International Conference on Retroviruses and Novel Drugs June 08-09, 2015 Chicago, Illinois, USA

Biography:

Abstract:

HIV infected long-term nonprogressors (LTNP) and elite controllers (EC) maintain enduring control of HIV infection without antiretroviral therapy (ART). Emerging evidence suggests that cells in tissues such as the gut continue to produce low-levels of HIV, low-level ongoing viral production occurs in the body even with concurrent ART. Whether LTNP/EC receiving ART can completely suppress low-level ongoing viral replication is unknown. We have previously found that SIV infection in Chinese rhesus macaque (ChRM) is the closest animal model that mimics HIV infection. We have also shown that gut mucosa is a major reservoir in LTNP/EC of SIV-infected ChRM. Here we studied SIV mutation and evolution in the gut-associated lymphoid tissue to determine whether ART can fully suppress viral replication in LTNP/EC of SIV-infected ChRM. Ten animals were treated with reverse integrase inhibitors (tenofovir and Emtricitabine) daily for up to 24 weeks. PBMCs and lymphocytes were isolated from blood, spleen, lymph nodes and gut tissues of 6 sites (duodenum, jejunum, ileum, cecum, colon and rectum) by the end of ART. Nested PCR and single genome amplification (SGA) was performed. SIV envelope gp120 gene from positive samples was amplified. Sequences from PBMC before and after ART were compared with sequences isolated from the 6 sites of the small and large intestine. Phylogenetic trees were constructed. Viral mutation and evolution were analyzed. The size of reservoirs in PBMC and lymph nodes was evaluated by quantitative coculture assays. We found that plasma viral loads (pVL) in all animals were suppressed and sustained below 28 copies/ml (limitation of detection) during ART. Two animals had viral rebound after approximately 2 weeks of cessation of ART. Viral DNA loads were detectable in either PBMCs and/or lymphocytes from different gut sections at variant levels by the end of ART. The frequency of detective PCR positives was higher in large intestine than small intestine, indicating large intestine is a major reservoir. A total of 245 sequences were analyzed. While envelope sequences showed limited viral mutations in some animals, viral evolution was observed in different sections of the gut in other animals even received 24 weeks of ART with sustained undetectable pVL. The size of viral reservoirs in blood reduced in LTNPs/ECs after ART. However, spleen and lymph nodes had higher resting CD4+ T cells carrying replication-competent SIV than blood by the end of ART. These data suggest that low-level of ongoing viral replication may still occur in some LTNP on ART. The SIV/ChRM nonhuman primate model is excellent for testing novel therapeutic interventions on typical progressors and LTNP/EC for HIV eradication.

Biography:

Lawrence Kleiman completed his Ph.D at Johns Hopkins University, and after postdoctoral studies at the Beatson Cancer Institute in Glasgow, Scotland, has spent the the remainder of his career at the Lady Davis Institute for Medical Research, Jewish General Hospital, in Montreal, Quebec, where he is also a Professor in the Department of Medicine at McGill University. He has published over 140 papers, and has worked for the last 20 years studying HIV-1 replication at the McGill AIDS Centre.

Abstract:

During HIV-1 infection, the conversion of the HIV-1 RNA genome into a double-stranded DNA that can be integrated into the host cell’s genome is accomplished by reverse transcription. The initiation of this reverse transcription requires a specific cellular tRNALys3 to act as a primer. This tRNA is annealed during assembly of new HIV-1, i.e., the infecting virus already contains annealed tRNALys3. The 3´ 18 nts of tRNALys3 anneal to a complementary 18nt sequences in the viral RNA termed the primer binding site (PBS), and in this presentation, we will discuss the cellular and viral factors that promote the ability of the tRNALys3 to locate the 18 nt sequence within the >9 KB comprising the HIV-1 viral RNA genome. This is associated with the formation of an early HIV-1 assembly intermediate at the site of the Gag/GagPol translation, whose components include Gag, GagPol, lysyl-tRNA synthetase (LysRS), and tRNALys3. This intermediate contains an increased concentration of tRNALys3 due to a specific interaction between viral Gag and LysRS, and is able to bind through LysRS to a viral RNA structure near the PBS that resembles tRNALys3 . This initial cytoplasmic annealing of tRNALys3 to viral RNA is part of a multi-staged annealing process, and not only functions in promoting reverse transcription, but also facilitates a conformation change in the 5´-untranslated region of viral RNA that promotes viral RNA dimerization.

Biography:

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Background: In Ethiopia, free access to ART has been expanded rapidly since 2005. With the rapid scale up and decentralization of the ART service, the emergency and transmission of HIV drug resistance (HIVDR) will be a major problem. In view of this, we evaluated the prevalence of transmitted HIVDR (TDR) in recently HIV-1 infected adults in Gondar town, Ethiopia using WHO threshold surveillance method. Methods: A cross-sectional survey was conducted among antiretroviral-naive adults who were seeking HIV diagnostic testing in VCT site in Gondar university hospital and Gondar Health center, from August 2011 to December 2012 using the WHO recommended eligibility criterion. A total of 84 study participants fulfilling the inclusion criterion were consecutively enrolled, and blood specimen were collected. HIVDR genotyping was done using the in-house assay. Sequence were interpreted using the calibrated population resistance (CPR) tool of the Stanford university database according to the 2009 WHO surveillance drug resistance mutation (SDRM) list. Result: Among the 84 participants, 70 (83.3%) were female and mean age was 21years (range: 18–24 years). Amplification and sequencing was successful for 67(79.8%) of specimens.Using the WHO-recommended truncated sequential sampling technique, among the first 47 sequenced specimens, 3 were found to harbor major HIVDR mutation associated to non-nucleoside reverse transcriptase inhibitor (NNRTI), suggesting moderate prevalence (5%–15%) of TDR in the study area. The mutation detected were K103N (n=2) and G190S (n=1). Including all sequenced sample in the analysis (n=67), 4(6%) were found to have major HIVDR mutations (K103N (n=2), G190S (n=1), and Y181Y (n=1)) associated to NNRTI. However, no nucleoside/tide reverse transcriptase inhibitor (NRTI) or protease inhibitor (PI) associated major HIVDR mutation were detected. Conclusion: In comparison to previous studies done in the Gondar town, our result showed an increase in prevalence of TDR, which could be associated to the ART scale up. As the TDR HIV-1 may seriously affect the efficacy of first-line ART, the moderate levels of TDR observed in this study area indicate the need for continuous surveillance of TDR in Gondar town and in different region of Ethiopia to optimize treatment efficacy of the current ART and improve the drug resistance strategy within a country.

Biography:

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Background: Many clinical trials[1, 2] have been carried out assessing the feasibility of cell phone text messaging to improve adherence to their medication and positive results have been reported; 12% improved adherence reported by the Lester et al study. However there was need to assess the perceptions of the HIV infected patients towards the use of these cell phones to support their adherence to medication which necessitated this study. Methods: A cross sectional survey was conducted among HIV infected patients receiving Highly Active Anti-retroviral Therapy at the Comprehensive Care Clinic in Kenyatta National Referral Hospital between May and July, 2011. Pre-tested questionnaires were used to collect the socio-demographic and perceptions data. The recruitment of the participants was done using the random probability sampling method and data analyzed using SPSS version 16.0. Results: A total of 500 HIV infected patients (Male-107, Female-307) aged 19-72 years were interviewed among which (99%) had access to cell phone and with support to the idea of using cell phone to support adherence. A greater proportion (46%) needed the cell phone to seek medical advice on factors hindering their adherence but only 3% needed it to receive reminders. Majority (72%) preferred calling the healthcare provider with their own phones and only 0.4% preferred to be called or texted by the doctor. Most (94%) of the participants had no problem with confidentiality infringement but (6%) of them had problem with it; majorly the young population. Conclusion: Doctor-patient communication via mobile phone is acceptable strategy to improve adherence as it offers the patient opportunity to seek medical advice by calling the doctor with their phones. Confidentiality infringement is only an issue among the young population calling for more studies to determine the associated factors to this behavior.

Biography:

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Colon cancer is the second leading cause of cancer related deaths in the United States. The nuclear factor B (NF-B) is an important family of transcription factors whose aberrant activation has been found in many types of cancer, including colon cancer. Therefore, understanding the regulation of NF-B is of ultimate importance for cancer therapy. The purpose of this study is to use a novel validation-based insertional mutagenesis (VBIM) strategy to identify novel regulators of NF-B, and further evaluate their roles in the regulation of NF-B signaling in colon cancer cells. We infected Z cells (293 derived cells with hyper active NF-B activity) with VBIM virus to cause the overexpression of negative regulators of NF-B, and then further selected the mutant cells with low NF-B activity under ganciclovir (GCV) treatment. Targeted gene was then identified by using VBIM specific primers. In a preliminary screen, we identified the novel adenomatous polyposis coli like protein (ALP) gene as a negative regulator of NF-B. Overexpression of ALP led to decreased NF-B activity by B reporter assay, while knocking it down had the opposite effect. Furthermore, we found that overexpression of ALP in HT29 colon cancer cells greatly reduced both the number and the size of colonies that were formed in a soft agar assay, while using shRNA resulted in an opposite effect, confirming that ALP is a tumor suppressor in HT29 cells. Future experiments aim to further assess the role of ALP in colon tumor formation in a mouse xenograft model. In summary, by using the novel VBIM technique, we identified ALP as a novel negative regulator of NF-B. This discovery could lead to the establishment of ALP as a potential biomarker and therapeutic target in colon cancer.

Biography:

Franck Mortreux has completed his PhD from Paris 6 University (France) and Postdoctoral studies from Agronomy Universirty of Gembloux (Belgium). He is PI in France in the Ecole Normal Superieur de Lyon (UMR5239, LBMC). His researches are mainly focused on HTLV-1 infection and on molecular mechanisms underlying post-transcriptional modifications, virus persistence and oncogenesis.

Abstract:

Reprogramming cellular gene transcription sustains HTLV-1 viral persistence that ultimately leads to the development of adult T-cell leukemia/lymphoma (ATLL). We hypothesized that besides these quantitative transcriptional effects, HTLV-1 qualitatively modifies the pattern of cellular gene expression. Exon expression analysis shows that patients’ untransformed and malignant HTLV-1+ CD4+ T-cells exhibit multiple alternate exon usage (AEU) events. These affect either transcriptionally modified or unmodified genes, culminate in ATLL, and unveil new functional pathways involved in cancer and cell cycle. Unsupervised hierarchical clustering of array data permitted to isolate exon expression patterns of 3977 exons that discriminate uninfected, infected, and transformed CD4+ T-cells. Furthermore, untransformed infected CD4+ clones and ATLL samples shared 486 exon modifications distributed in 320 genes, thereby indicating a role of AEUs in HTLV-1 leukemogenesis. Exposing cells to splicing modulators revealed that Sudemycin E reduces cell viability of HTLV-1 transformed cells without affecting primary control CD4+ cells and HTLV-1 negative cell lines, suggesting that the huge excess of AEU might provide news targets for treating ATLL. Taken together, these data reveal that HTLV-1 significantly modifies the structure of cellular transcripts and unmask new putative leukemogenic pathways and possible therapeutic targets.

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Glioblastoma multiforme (GBM) is considered to be one of the deadliest human cancers, characterized by a high proliferative rate, aggressive invasiveness and insensitivity to radio- and chemotherapy, as well as average survival is less than one year. On the basis of published data, there is sufficient evidence to conclude that HCMV DNA, mRNA, and/or antigens exist in most glioblastoma tissues. HCMV could modulate the malignant phenotype in glioblastomas by interacting with key signaling pathways that involve in apoptosis, angiogenesis, invasion and immunoevasion; including PDGFR, PI3K/AKT, STAT3 and GSK-3b. Existing evidence supports an oncomodulatory role for HCMV in glioblastoma, in this study need to focus on determining the role of HCMV as a glioblastoma-initiating event inPI3K/AKT pathway. EZH2 is a marker of PI3K/AKT pathway so we decided to evaluate the expression of this gene in 3 groups: Negative HCMV glioblastoma multiformetissus, positive HCMV glioblastoma multiformetissus and non tumor tissues. The presence of Human Cytomegalovirus was assessed according to our previous article. Human Cytomegalovirus was present in 75% of glioblastoma tissues. Then RNA was extracted, cDNA was synthesis and Real-time PCR was performed. Then rate of increased expression was calculated using the Livac or 2-ΔΔCt.ΔCt of samples in the three groups were compared using ANOVA (Analysis of Variance). Expression of EZH2 gene relative to GAPDH gene in HCMV negative glioblastoma tissues were increased 6.053 times compared to non-neoplastic brain tissues. Expression of EZH2 gene relative to GAPDH gene in HCMV positive glioblastoma tissues were increased 41.098 times compared to non-neoplastic brain tissues.ANOVA test showed thatthe difference of mean ΔCt for EZH2 gene between healthy subjects and patients with HCMV positive glioblastoma tumor and HCMV negative glioblastoma tumor is statistically significant (p-Value = 0.0001).

Biography:

Abstract:

Background: In Ethiopia where there is no strong surveillance system and diagnostic facilities are limited, the real burden of tuberculosis (TB) lymphadenitis is not well known. Therefore, we conducted a study to estimate the prevalence of TB lymphadenitis in Southwest Ethiopia. Methods: A community based cross-sectional study was conducted from February to March 2009 in the Gilgel Gibe field research area. A total of 30,040 individuals 15 years or older in 10,882 households were screened for TB lymphadenitis. Any individual 15 years or older with lumps in the neck, armpits or groin up on interview were considered TB lymphadenitis suspect. The diagnosis of TB lymphadenitis was established when acid fast bacilli (AFB) smear microscopy of fine needle aspiration (FNA) sample, culture or cytology suggested TB. HIV counseling and testing was offered to all TB lymphadenitis suspects. Descriptive and bivariate analysis was done using SPSS version 15. Results: Complete data were available for 27,597 individuals. A total of 87 TB lymphadenitis suspects were identified. Most of the TB lymphadenitis suspects were females (72.4%). Sixteen cases of TB lymphadenitis were confirmed. The prevalence of TB lymphadenitis was thus 58.0 per 100,000 people (16/27,597) (95% CI 35.7-94.2). Individuals who had a contact history with chronic coughers (OR 5.58, 95% CI 1.23-25.43) were more likely to have TB lymphadenitis. Lymph nodes with caseous FNA were more likely to be positive for TB lymphadenitis (OR 5.46, 95% CI 1.69-17.61). Conclusion: The prevalence of TB lymphadenitis in Gilgel Gibe is similar with the WHO estimates for Ethiopia. Screening of TB lymphadenitis particularly for family members who have contact with chronic coughers is recommended. Health extension workers could be trained to screen and refer TB lymphadenitis suspects using simple methods.

Biography:

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Background: Rotavirus infections are a major cause of diarrhea in children under 5 years in both the developed and developing countries. Rotavirus is often associated with acute infection with high severity level that causes death. Genetic rotavirus, patient immunity, and environmental are thought to be related to the severity of the incidence of acute diarrhea due to rotavirus in infants and young children. Objective: To prove correlation rotavirus genotypes and severity degree of acute diarrhea. Methods: Cross sectional study was conducted in children aged 1-60 months with acute diarrhea hospitalized at gastroenterology ward Dr. Soetomo Hospital between January to June 2014. Examination of rotavirus in stool specimens made from bed-side examination using BD-Rota/Adeno Examine kit, whereas rotavirus genotypes examined by molecular methods, namely reserve transcription-polymerase chain reaction (RT-PCR) two-step at the in situ of Tropical Disease (ITD) University Airlangga by personnel trained analysts at ITD. The severity of diarrhea was measured by using a scoring system scores Ruuska and Vesikari (1990). Result: A total of 88 children met the criteria, 80.7% were aged 6-24 months. Average value of the total score of severity of diarrhea was 10.21 (SD 2.12). Most rotavirus genotype G2P {4} (19.3%) to group common genotype and genotype G1P {4} and G9P {4} for group genotyping uncommon with a prevalence of respectively 4.5% . There are significant differences between the types common genotype and uncommon genotype to the total score of the severity degree of diarrhea (p<0.05) Conclusion: Severity degree of acute diarrhea in children with genotype G2P {4} is the highest prevalence genotype of rotavirus in East Java.

Biography:

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Background: Hepatitis is an inflammatory condition of the liver and viral hepatitis is a conventional term used to denote hepatitis caused by hepatotrophic viruses (Hepatitis A-G). High prevalence of these viruses are reported in Nigeria. Hepatitis B and C may cause liver cirrhosis and they can be contacted through contaminated blood and blood products. Many blood banks in Nigeria screen for hepatitis B and C using immune-chromatographic screening method (Rapid test strip). This is because these strips are readily available in the market, cheap, requires no electricity for storage, special training or equipment before use. The intent of our study is to compare the sensitivity of this method using an advanced immunological method. Method: 660 potential donors are tested for hepatitis B surface antigen (HBs Ag) and hepatitis C virus antibody using immune-chromatographic test strip and ELISA methods. Result: We found out that 38 (5.7%) out of 660 subjects tested positive for HBS Ag using immunochromographic method while 71 (10.8%) were positive using ELISA. None is positive for hepatitis C antibody using immunochromatographic method while 4 (0.6%) subjects were positive using ELISA method. Conclusion: Immunochromatographic method is not good enough to screen blood donors for hepatitis B and C.

Biography:

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HIV Viral infectivity factor (Vif) evokes the destruction of the host restriction factor known as APOBEC3G (A3G). Vif dimerization has been shown to be essential for Vif binding to A3G and A3G degradation. Experimental data show that in the absence of Vif, virion-­‐assembled A3G will hypermutate proviral DNA during reverse transcription. The open questions are: can Vif be successfully drugged and by sparing A3G, will hypermutation activity be sufficient to inhibit viral replication? High throughput screening (HTS) assays were developed using FRET for Vif dimerization. The screens were used to select compounds with dose-­‐dependent signals by preventing Vif FRET. A secondary assay for Vif-­‐dependent A3G degradation was used to identify compounds that preserved A3G. These were triaged for the ability to inhibit pseudotyped HIV replication and to have low cytoxicity. Ion torrent sequencing of integrated viral genomes revealed extensive hypermutation characteristic of A3G preferences. Following medicinal chemistry, a lead was tested in a seven day spreading infection in PBMC. The Vif dimerization FRET assay provided a robust HTS method applicable to a large library of drug-­‐like molecules. Quantitative HTS followed by orthogonal secondary screen and cytotox counter screening enabled selection of limited number of chemistries for pseudo typed viral testing. A lead (SMVDA) was selected with nanomolar IC50. By impairing Vif dimerization, A3G degradation was reduced and viral particle incorporation of A3G was enhanced. Proviral DNA isolated form target cells showed numerous tracts of dG to dA hypermutation that corresponded to multiple nonsense codon and sense changes. 17 different clades and 8 drug resistant strains of HIV infecting PBMC were sterilized by seven days following a single dose of SMVDA . Conclusions: Drugging Vif led to massive dG to dA hypermutation of HIV proviral DNA, such that the protein coding capacity of the virus would be severely compromised. Inhibitors of Vif are broadly neutralizing and inhibited all drug resistant strains of HIV tested. Drugging Vif therefore is achievable and the data suggest that this will serve as a firewall for viremia induced by activating reservoirs as well as a solution for rescue and salvage therapies.

Biography:

Luise Mansky is Professor and Director of the Institute for Molecular Virology, University of Minnesota. He received his PhD degree in Molecular Virology from Iowa State University and was a postdoctoral fellow in the laboratory of Dr. Howard Temin at the McArdle Laboratory for Cancer Research, University of Wisconsin-Madison.

Abstract:

Multiple therapeutic approaches are being investigated towards eradicating viral reservoirs towards a cure to HIV-1 infection. Our current studies have been directed towards the discovery of viral mutagens that can eliminate the infectivity of reactivated virus through elevation of mutational load. We have found that 5-azacytidine and the combination of decitabine and gemcitabine work as potent viral mutagens and progressed in the preclinical evaluation of these antiretroviral strategies. We have discovered new antiretrovirals that also have potential for such therapeutic applications, which will be discussed

Biography:

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Background: Though HIV/AIDS is one of the worst health crises in the recorded history of the world, it has moved beyond being primarily a health issue, to developmental crisis. More than 83% of all new infections in many African countries are among young people. Higher learning institutions need to seriously address HIV/AIDS in their mandate. This study sought to assess risk perception, HIV/AIDS related knowledge, attitude and behaviors of the ECSC community. Materials and methods: A cross-sectional survey was conducted between January and June 2011.Mixed methods of combining structured questionnaire, focus group discussion and key informants interviews were conducted. Results: Out of 250 respondents, 238 returned the questionnaire. The majority of respondents know about the risk, ways of transmission and prevention about HIV and AIDS. All respondents never perceive they are at risk for HIV. Unsafe sex and multiple concurrent sexual partnerships were found among all religious groups, married staff and students. Qualitative data support this finding. Conclusions: Positive changes in awareness and attitudes toward HIV/AIDS were noted, yet comprehensive knowledge is lacking. An improved strategy to promote comprehensive knowledge and behavioral change interventions is needed in ECSC. Implications from these finding suggest that other institutions may benefit from similar changes.

Biography:

Rafael Correa Rocha has completed his first PhD in Biology at the Universidad Complutense of Madrid (Spain) in 2004. His thesis was awarded with the National Prize of Doctorate 2005. He joined the ISREC (Epalinges, Switzerand) as a Post-doctoral researcher and afterwards, he obtained a position as Assistant Professor at the Hopitaux Universitaires de Genève (Switzerland). He joined the IISGM of Madrid as a Senior Scientist in February 2008. He completed a second PhD in Medicine at the Universidad Autónoma of Madrid in 2014. At present, he is the Head of the Laboratory of Immune-regulation at IISGM. He has published more than 40 papers in reputed journals.

Abstract:

Regulatory T cells (Treg) play an important role in infections modulating host immune responses and avoiding over-reactive immunity. Immune hyperactivation associated with HIV infection lead to a marked erosion and deregulation of immune system, and by that, the role of Treg in HIV-infected patients is critical because their implication preventing this hyperactivation. The findings about the role of Treg in HIV infection are controversial, and considering that Treg are susceptible of being infected by HIV, there are not data about the effect of HIV infection on Treg phenotype and function.We demonstrated by first time that HIV infection of Treg markedly disturb the phenotype of these cells downregulating the expression of Foxp3 and CD25, which is followed by a loss of their suppressive capacity. We also demonstrated that the balance between Treg and effector cells is broken in HIV patients by a direct effect of the virus on Treg, and finally we have also described that HIV-infected patients have a marked deficit and impaired function of Treg that would be related with the incidence of Immune hyperactivation in these patients.

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Human Immunodeficiency Virus (HIV) uses the human CD4 protein (hCD4) as the primary surface receptor for attachment and infection of host cells such as CD4+Th-lymphocytes. This hCD4 receptor dependency can be exploited in an antiviral strategy: Removal of the CD4 receptor from the cell surface will prevent HIV infection of target cells and viral replication.In eukaryotic cells, surface expression of most transmembrane proteins is dependent on the presence of a hydrophobic N-terminal signal peptide (SP) on nascent proteins. It facilitates targeting of the nascent proteins to the Sec61 translocon, a universally conserved protein-conducting channel in the ER-membrane, and subsequent insertion of the chain for translocation. Despite their common function, signal peptides have diverse primary sequences. Thus, drugs that recognize unique signal peptide sequences could be exploited to inhibit translocation of selected proteins into the ER and their expression at the cell surface. Previously, the small-molecule macrocycle CADA was identified as an antiviral drug with broad spectrum anti-HIV activity. It acts as a highly selective hCD4 expression down-modulator. Here we show that CADA inhibits hCD4 biogenesis by preventing co-translational translocation of hCD4 to the ER lumen, both in cell culture and in a cell-free in vitro translation/translocation system. The activity of CADA maps to the signal peptide of hCD4 which represents the minimal sequence requested for full CADA sensitivity. Importantly, we could show direct binding between this SP and CADA through suface plasmon resonance (SPR). Furthermore, translocation inhibition by CADA causes the precursor protein to be routed to the cytosol for degradation. These findings demonstrate that a synthetic, cell-permeable small-molecule such as CADA can act as a signal peptide-binding drug to regulate the expression of specific target proteins by selective and reversible inhibition of protein translocation.

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Small molecules targeting the enzymes responsible for human immunodeficiency virus (HIV) polyprotein maturation (protease), DNA synthesis (reverse transcriptase) and the subsequent insertion of ribonucleotide-free double-stranded DNA into the host chromosome (integrase) have for several years been the central components of combination antiretroviral therapy. For infected individuals harboring drug-susceptible virus, this approach has afforded complete or near-complete viral suppression. However, in the absence of a curative strategy, the predictable emergence of drug-resistant variants requires continued development of improved antiviral strategies, inherent to which is the need to identify novel targets. Cis-acting regulatory elements of the HIV-1 RNA genome that regulate its transcription (the transactivation response element, TAR), translation (the ribosomal frameshift signal), nucleocytoplasmic transport (the Rev response element or RRE), dimerization (the dimer linkage sequence or DLS), packaging (the  element) and reverse transcription of the (+) strand RNA genome (the primer binding site, or PBS) should now be considered as alternative targets for small molecule, peptide- and oligonucleotide-based therapeutics, as well as combinations thereof. The first part of this talk will summarize how high-resolution 3D structural information is being used to develop small molecule and peptide-based therapeutics that target critical cis-acting RNA motifs of the HIV-1 genome and consequently may be less prone to resistance-conferring mutations. Subsequently, advances in the development of novel high-throughput small molecule microarrays (SMMs) and RNA motifs that have been successfully targeted by this approach will be presented. An extension of the (SMM) approach to target other viral RNAs, or virus-specified RNAs, will be presented. Finally, where target specificity, endosomal release, cellular penetration and toxicity have been the primary obstacle to successful “macromolecule therapeutics”, methodological advances will be reviewed.

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Background: Client satisfaction and adherence on ART service were an important task for care providers to increase service utilization and to respond to HIV emergency; however, Other provider-defined criteria is far from ideal if as a result of the service that the patient is unhappy or dissatisfied. There is, then, a sound rationale for making the organization and delivery of health service responsive to consumer opinion. Objective: The aim of this study is to assess client satisfaction and adherence on an ART service provision in Jimma University specialized hospital. Methods: A cross sectional study involving both quantitative and qualitative data collection methods was conducted from may 1-30/2010.A total of 337 Adult PLWHA on ART for at least 3 months were the study participants. Systematic sampling technique was used to select the study subjects. Data were collected using structure questionnaire, check lists and semi structure interview guide. After clearing and checking for consistency data were coded, entered and univariate and multivariate analysis was carried out using SPSS version 16.0. qualitative data‘s were transcribed and narrated under themes. Result: A total response rate of 100% from 337 sample size was obtained. Among those 203 (60.2%) were females. Two hundred thirty one (68.5%) of respondents scores  mean which means 68.5% satisfied relationship with their care providers. In this study, in which adherence was measured using a self report method, 95.5% of patients were adherent with 95% prescribed doses. Marital status, occupation, and waiting time were was found to be associated significantly with adherence [OR.136, 95%CI.019-.997], [OR 9.341, 95%CI 1.189-73.358], [OR 9.88E-24, 95%CI 1.759E-24-5.550E-23] respectively. Conclusion and Recommendation: The result of the study showed that assessment of client satisfaction and adherence in Jimma University specialized hospital is high adherence rate in spite of satisfaction. Overall client satisfaction and patient, provider relationship satisfaction rate were low. However measured by self report method, adherence to ARV treatment in this study was seems to be encouraging. Working with other religious leaders, and community leaders to strengthen adherence status are recommended.

Biography:

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Background: Highly active antiretroviral therapy is the cornerstone of management of patients with human immunodeficiency virus infection. Antiretroviral therapy can prolong survival of patients however this drugs are associated with adverse effects that can affect patient adherence and if severe may require regimen change. The aim of the study is to assess the prevalence of antiretroviral associated adverse effects and management strategies undertaken among patients taking antiretroviral therapy in Jimma University Specialized Hospital. Methods: A retrospective review of patient medical records (2009-2011) was done to assess adverse effects associated with antiretroviral therapy. A sample of 403 patient medical records was selected using systematic random sampling method. Data was collected using structured data abstraction format. Data were entered into SPSS windows version 16 and chi-square test was used to analyze factors associated with adverse effects. P-value of less than 0.05 was considered as statistical significant. Results: About 65.5% of patients had developed at least one adverse effect to antiretroviral drugs. The most commonly encountered adverse effects were gastrointestinal and central nervous system effects. Severe side effects that resulted in high rate of regimen switch and discontinuation included anemia, peripheral neuropathy, rash and hepatotoxicity. Conclusion: Majority of patients taking antiretroviral therapy experienced mild to severe adverse effects in the course of treatment which can affect the patient treatment outcome. Thus close monitoring of toxicities considering the risk-benefit ratio of continuing, switching or discontinuation of treatment is critical.

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Background: Despite the massive resources and intensified interventions, desired decline in HIV/AIDS epidemics has not been achieved. This study was aimed to evaluate how students are reacting being faithfulness message for HIV/AIDS prevention using Extended Parallel Process Model (EPPM). Method: Cross sectional study was conducted using quantitative and qualitative methods of data collection. Structured self-administered questionnaires were used to collect data. Simple random sampling was used to select respondents. Quantitative data were analyzed using SPSS version 16.0. Qualitative data were analyzed using OpenCode software. Results: 61.5% (251/408) of the respondents were in danger control response whereas 38.5% (157/408) of the respondents were in fear control response. As independent predictors, self efficacy [AOR (95%CI)=0.32 (0.37 to 0.72)], response efficacy [AOR (95%CI)=0.82 (0.59 to 0.98)] of HIV/AIDS, religion (catholic) [AOR (95%CI)=0.33 (0.65 to 0.69)] and age (20-24) [AOR (95%CI)=0.13 (0.43 to 0.73)] were positively associated with danger control response where as fathers’ occupation [AOR (95%CI)=3.31 (5.55 to 19.08)], perceived susceptibility to [AOR (95%CI)=4.42 (2.44 to 8.61)], perceived severity of [AOR (95%CI)=5.33 (3.21 to 14.74)] HIV/AIDS and not hearing faithfulness message [AOR (95%CI)=5.11 (6.91 to 17.08)] were negatively associated with danger control response. The EPPM Model explained 59.04% of variance in this study. Conclusion: Despite higher numbers of students were in danger control psychological responses, intolerable numbers are in fear control responses. Therefore, due attention should be given to fill the gap of perception of risk in both self-efficacy to be fixed with one sexual partner and response efficacy to be stayed with that one sexual partner tailoring the message in the context of their religion, age and their income.

Biography:

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Background: Mother-to-child transmission is by far the largest source of HIV infection in children below the age of 15 year and accounts for 90% of childhood HIV infections. The transmission of HIV from infected mothers to babies could occur during pregnancy, delivery and breastfeeding. For women to take advantage of measures to reduce transmission, they need to know their HIV status. Objective: To assess the knowledge, attitude and practices towards prevention of mother-to-child transmission of HIV/AIDS and associated factors among pregnant women in Hossana town Methods: community based cross-sectional study was conducted at Hossana town from November 2013- December 2014. Of a total of 422 sample women, 417 pregnant women in Hossana town was included in the study and data were collected by using pre tested structured questionnaire. The collected data were cleaned, edited, fed to computer to be analyzed using SPSS window version 16.0. Results were presented by ratios, tables & diagrams. Binary and multiple logistic regression analysis were done to identify factors associated with KAP towards PMTCT. Result: A total of 417 pregnant women responded to the questionnaire, yielding a response rate of 98.8%.78.2% knew that the risk of MTCT of HIV can be prevented. 69.92% responded that ART should be started as soon as the status known. 64% of the respondents had good knowledge (greater than average score) on PMTCT. Majority, 98.3% of the respondents had good attitude. 90.4% respondents were practiced PMTCT of HIV. Most, 90.4% mothers tested for HIV during current pregnancy and 93.9% shared test result to their husbands/partners. Husband’s attended higher education, family size of 9-11, Place of the most recent child delivery being health facility, ANC visit for current pregnancy, Awared on HIV counselling and testing, Participated during community conversation on HIV/AIDS issues had shown significant association with knowledge of PMTCT of HIV. On the other hand, Knowledge of MTCT of HIV had shown significant association with practice of PMTCT of HIV. Conclusion: More than three-forth (78.2%) mothers knew about PMTCT of HIV and 44.7% knew ART drugs could reduce risk of MTCT of HIV. Majority, 98.3% had good attitude towards PMTCT of HIV. Most, 90.4% mothers tested for HIV during current pregnancy and 93.9% shared test result to their husbands/partners. Recommendation: Awareness creation activities on HIV/AIDS in general and prevention of MTCT in particular through distribution of ICE/BCC materials, mass medias and maternal and child health service units should be strengthened.

Biography:

Stephen Talugende completed an MPH from the University of Liverppol, and is currently DrPH candidate at Walden University. He is the Chief HIV/AIDS Officer of the United Nations Interim Force in Lebanon. Stephen is a self-made adaptive leader that developed from a service recipient to a public health professional. As a result, he was recognised as the Highly Commended Outstanding Leader of the year 2013 by the British National Leadership and Management awards in association with the Chartered Management Institute for his extensive work on HIV and AIDS programs management, advocacy, and community mobilization, capacity building, training, counseling, social and behavior change communication, etc. He has held several portfolios, including Chief Executive Officer of the National Forum of PLHIV networks in Uganda, Africa Representative for Mildmay International, and HIV/AIDS Policy Advisor to the United Nations Operations in Burundi, and carried out many technical support and consultancy services.

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Protracted expectations from HIV research and treatment has been and continue to be giving hope to multidisciplinary teams and holistic interventions for prevention and impact mitigation of the pandemic. Whilst comprehensive resources are committed to generating appropriate knowledge through research, empathetic ethical practice epitomises holistic needs of beneficiaries and service providers. HIV shatters not only dreams of those infected, but also visions for the future of possibilities for improved health and related socioeconomic services. Requisite outcome of both research and treatment is when knowledge and hope becomes pragmatic, reflects more intrinsic community perspectives, and easily traslates community perspectives, meaning and conclusions. Communities, according to Bacon (2009), due to their unity, they share common beliefs and interests. Consequently, the threshold of production, publication, availability and usage of research and treatment is based on empathetically identifying how peoeple’s needs affect their ability to achieve relevant outcomes, and how this impacts on their wellbeing, as well as consistent effectiveness of innovations. Phenominally, it is an ambivalent dimension to seperate researchers as professionals from researchers as social beings. Experientially, there is a critical missing link in the way research and treatment are considered from professional and scholarly practice. Thwaites (2007) attributed the model of therapeutic empathy within contexts of: empathic attunement, empathic attitude/stance, empathic communication, and empathy knowledge. It is not a matter of research and making treatment available, but how much professionals can practically relate to the products as prospective users. Years as a service recipient enabled me to recognise how the lack of empathetic relationship of professionals led to severe backdrops to shortcomings in practice. It is easy to see possibilities in the eyes of a researcher, but how easy is it to see possibilities in those of recipients. Conclusions are easier understood by professional, than they’re in those of communities. Therefore, research conclusions and most importantly treatment are always a major breakthrough, but are always a mere step in responses against pandemics like HIV. Empathy should, in that sense remain an integral component to any kind of research.

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Introduction: Initial entry of HIV-1 into host cells remains a compelling and yet elusive target for developing agents to prevent infection. This step is mediated by a sequence of interactions of a trimeric gp120/gp41 envelope (Env) protein complex with host cells, including initial gp120 encounter with the cellular receptor CD4 and a chemokine co-receptor usually either CCR5 or CXCR4. A peptide triazole class of entry inhibitor leads has been shown to bind to gp120 with close to nanomolar affinity, to suppress protein ligand interactions of the Env protein at both its CD4 and co-receptor binding sites and to inhibit cell infection by a broad range of virus subtypes. Further we have also shown that gold nanoparticle conjugated peptide triazoles lead to 20 fold enhanced potency of their anti-viral effects against HIV-1. Previously bowman et al. has shown that multivalent display of HIV inhibitors on gold nanoparticles (AuNPs) has lead to a substantial amount of poency enhancement. This study will lead to the study of size dependency and density dependency on the gold nanoparticle peptide triazole conjugates to further understanding of their mechanism of action leading to enhanced potency. Materials and Methods: The AuNPs were synthesized using a modified citrate reduction method to obtain size-controlled, stable and monodisperse AuNPs. The peptide (KR13) was conjugated to the AuNP using a direct gold-thiol covalent link. The size and extent of polydispersity of the AuNP-KR13 conjugates were measured using Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS). The HIV-1 viral entry inhibition potencies of KR13 and AuNP-KR13 conjugates were compared using a single-round pseudoviral infection luciferase reporter assay using lab synthesized pseudoviruses. We subsequently tested the effects of KR13 and AuNP-KR13 on the virus particle itself by measuring release of the nucleocapsid protein p24 using ELISA analysis of cell-free virion inhibition. Results and Discussion: Compared to peptide triazole alone, the 20 nm AuNP-KR13 conjugate exhibited a close to 20-orders of magnitude enhancement of infection inhibition activity (Table) and further with increasing size of AuNP, the potency was enhanced further with the 120 nm AuNP-KR13 having nearly 1600 fold enhancement (Table). KR13 and AuNP-KR13 conjugates are specific for HIV-1 envelope.There was no significant in vitro cytotoxicity observed for either KR13 or AuNP-KR13 conjugates. Further the residual virion that was generated upon treatment with KR13 was immunoreactive while the residual virion generated upon AuNP-KR13 treatment was completely inactive. The mechanism was studied by observing the morphological changes caused by both KR13 and AuNP-KR13 on the HIV-1 virion using TEM.

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Recently, WHO have reported that 35.3 million people were living with HIV/AIDS (PLWH) globally at the end of 2012 which included about 0.8% of adults aged 15-49 years. National AIDS Control Organization (NACO) of India has shown the prevalence of AIDS in India in 2013 to be 0.27 million. In the low and middle income countries with resource limited settings, more than 8 million PLWH were receiving antiretroviral therapy (ART) at the end of 2011. Application of highly active antiretroviral therapy (HAART) worldwide has been able to significantly reduce the mortality and morbidity of human immunodeficiency virus type 1 (HIV-1) infected individuals. However, the appearance of clinical drug resistance in AIDS patients due to various factors including nonadherance to medication (intake of antiretroviral) has been found to be associated to chemotherapeutic and virologic failure. In addition, high rate of viral replication, appearance of heterogenous circulating viral quasispecies, infidelity in proviral cDNA synthesis as well as immunological and pharmacological factors are also associated to drug resistance. The present research paper describes our understanding on the mechanistic aspect of drug resistance in HIV-1 against the application of the second generation antiretrovirals and possible strategies to encounter the issue. The recent approaches to explore new targets for actions for specific actions of the antiretrovirals and to develop target-structure based new small molecules with least toxicity to the host would be discussed.

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Background: BST-2(Tetherin/CD317) has been recently recognized as a potent restriction factor that inhibits the release of HIV-1 particles from infected cells. HIV-1 Vpu induces the degradation of BST-2 and allows virus escape from the restriction. Therefore, blocking Vpu-mediated down-regulation of cell surface BST-2 provides clearly an opportunity for developing anti-HIV-1 drugs. Method: In this study, a cell-based ELISA assay was used to screen for small molecule compounds that restore cell surface BST-2 in the presence of Vpu. Co-IP, BRET, cell image and other techniques were performed to study the mechanism of positive compounds. Results: In this study, we identified two small molecule compounds IMB-AZ and IMB-LA, both of which can restore cell surface BST-2 in a Vpu-dependent manner. Interestingly, the compounds had no effect on the degradation of IFNAR1 induced by IFN, suggesting that their abilities to block BST-2 degradation most unlikely resulted from a general inhibition of lysosome degradation pathway. Mechanism study revealed that IMB-AZ specifically inhibit Vpu-induced ubiquitination of BST-2 without interrupting the formation of Vpu-β-TrCP complex. Unlike IMB-AZ, the treatment of IMB-LA resulted in accumulation of BST-2 within CD63-containing endosomal compartments, suggesting that IMB-LA may impair the translocalization of BST-2 to lysosomes，and consequently inhibit the degradation of BST-2. Furthermore, the results of viral infectivity assay demonstrated that both of IMB-AZ and IMB-LA inhibit the release of HIV-1 particles and the infectivity of progeny virions in cell lines expressing BST-2. Conclusion: In summary, results herein demonstrated that both of IMB-AZ and IMB-LA could specifically inhibit the degradation of BST-2 induced by Vpu, and impair HIV-1 replication in a BST-2 dependent manner, suggesting IMB-AZ and IMB-LA have potent potential to develop a new anti-HIV drug.

Biography:

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Insect hemolymph studies demonstrated the presence of active principles such as antiviral. The aim of this work was cloning, expression and concentration of the recAVLOEc protein with antiviral activity from L. obliqua hemolymph. For that, a PCR with specific primers for the antiviral protein, based on the sequence previously cloned in bac-to-bac system, was made. Restriction sites were inserted in the primer for connection to the plasmid pET28a. First reaction was performed with the restriction enzyme BamHI, followed by digestion with the enzyme HindIII. After binding insert in the vector, the construction was selected in E. coli BL21 (DE3) pLysS and the cloning was confirmed by sequencing. The protein expression was induced with IPTG (1mM) at 0.7 DO at 37°C for 4h. In order to concentrate the target-protein, precipitation by salting-out method was used. Initially, ultrasound bacterial lysis was performed. After centrifugation (8,000 rpm x 10min), the saturated salt solution (sodium or ammonium sulfate) was added to the supernatant phase of cellular lysate. This solution was maintained for 12h at 4°C until the formation of the precipitate phase. The solubility curve of recAVLOEc was determined by measuring the protein composition of the supernatant phase. The effectiveness of salts as precipitant agents was verified. In addition, antiviral tests were performed and a titration was made with Measles virus infecting VERO cells, in which 24µg recAVLOEc/mL resulted in the viral title 256-fold lower than control.

Biography:

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The aim of this study is to report the comparative results of using combined antiviral drugs on naturally-infected FIV cats in the asymptomatic and AIDS stages of the disease. The drug combinations evaluated were: ZDV alone and ZDV+3TC+Nevirapine. Plasma viral load was measured at the beginning of treatment and at one year. Total leucocytes, neutrophils and lymphocyte counts, CD4+/CD8+ ratios and biochemical profile values were also analyzed. Neurological disease was evaluated by evoked potentials determinations. The results showed a better efficacy of the combined therapy vs. ZDV alone, reducing the viral load, increasing the CD4+/CD8+ ratio and producing the improvement of the clinical signs in the asymptomatic stage. In the AIDS stage the differences between both protocols were not so marked, but the combined therapy produced better results compared with ZDV alone. The auditory and visual evoked potentials resulted good as predictors of the neurological disease and showing the evolution of the patients with both protocols contrasted.

Biography:

Ekwu B B Ochigbo is a Registered Pharmacist and a HIV & AIDS resource person. He holds an MPhil with distinction in HIV & AIDS Management from the Africa Centre for HIV & AIDS Management, Stellenbosch University, Cape Town, South Africa. He is also the Director of TibePharmacare, a pharmaceutical and health research consultancy close corporation. His interests are in therapeutics and treatment outcomes research.

Abstract:

The purpose of this research was to determine the level of adherence among HIV infected pregnant women on prevention of mother to child transmission (PMTCT) antiretroviral therapy, and to establish the factors that contribute to poor adherence and their relative importance, in order to suggest intervention strategies that will improve treatment adherence among this population. The study was conducted in Area W Clinic, Francistown Botswana, and was a prospective cross sectional study using semi-structured questionnaire, and data collection form. In total, 61 pregnant women participated in the study and were all within three to nine months gestation, and had been enrolled into the PMTCT program at least more than one month previously. The following were their characteristics: 75% were within the ages of 26 to 42 years old, 90% were single, 81% had attained secondary school education, and 60% were unemployed. Adherence was considered optimum if greater than or equal to 95%. The participants demonstrated a good knowledge of the importance of PMTCT treatment adherence. Reported optimum adherence levels were 84% by virtual analogue assessment, and 82% by pill count. Ninety eight percent of participants reported they did not miss any dose during the last three days before the interview. The most important factors influencing adherence from the study were pregnancy related illnesses, medication side effects, and month of pregnancy of the patient as participants tended to adhere less as they got closer to delivery. It is therefore important for care-givers to carefully monitor patients for these effects, and to carry out continuous adherence counselling with special attention given to those approaching delivery in order to improve or maintain overall adherence to PMTCT therapy. In conclusion, adherence levels to PMTCT therapy among the population sampled was high but can be further enhanced with interventions designed to cover and improve the highlighted areas in the implementation of the preventive therapy.

Biography:

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Background: Mother-to-child HIV transmission remains very high in Cameroon. Therefore follow-up of numerous HIV-infected infants is a critical issue in the country. Here, we investigated the file of HIV-infected infants remaining asymptomatic in the absence of anti-retroviral therapy (ART). The first goal was to obtain an estimate of the prevalence of infants with an HIV controller like status. Method: HIV-infected infants, aged 6 months to 17 years children presenting at CIRCB for biological examinations were enrolled upon signed a proxy-consent. The enrollment took place from April 2011 to February 2013. From the medical file of 359 HIV vertically-infected infants, 41 were found naive of anti-retroviral therapy and free of clinical symptoms. Diseases related to HIV infection (oral candidosis, zona, chronic diarrhea, pulmonary tuberculosis, dermatitis) were more particularly checked and corresponding infants not included in the study. From the selected infants, CD4 counts and viral load were recorded. Non-exposed children were enrolled as control group. Results: Of the 359 infants, 41 were ARV-naive and free from HIV clinical symptoms. Five of them (12%) exhibit a viral load < 1200 RNA copies/ml. Their CD4 counts were found not statistically different from those of a control group of HIV negative infants (p = 0.33). Furthermore, ten years after contamination, three children did exhibit a viral load < 5500 RNA copies/ml. Altogether, this suggests the existence of pediatric HIV controllers (pHIC) with a frequency much higher (>10%). Conclusion: Our preliminary cross-sectional study highly suggests the existence of pediatric HIV controllers like in Cameroon despite all disfavoring living conditions. However, a longitudinal study would be required to confirm this hypothesis. The development of an HIV vaccine applicable to infants of countries with high incidence of HIV-infected people should benefit from the immunological analysis explaining the HIV controller (HIC) status.

Biography:

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Introduction: There is a growing concern about the increasing rates of Loss to Follow up (LTFU) among people who are on HIV/AIDS treatment programs. It is more common in resource-poor settings. However little is known about the time to LTFU and predictors after Antiretroviral Therapy (ART) initiation in low resource settings including Ethiopia. Method: Retrospective cohort study was employed among a total of 520 records of patients who were enrolled on antiretroviral therapy in Aksum St. Marry hospital. Baseline patient records were extracted from electronic and paper based medical records database and analyzed using Kaplan Meier survival and Cox proportional hazard model to identify the independent predictors of loss to follow up of patients on ART. Result: of 520 patients, 51(9.8%) were loss giving a LTFU rate of 8.2 per 100 person- years. From these LTFU, 21(41%) occurred within the first Six months of ART initiation. The independent predictors of LTFU of patient were being smear positive pulmonary TB [Adj.HR (95% CI)=(2.05 (1.02, 4.12)], male gender [Adj. HR (95%CI)=(2.73 (1.31, 5.66)], regiment AZT-3TC-NVP [Adj. HR (95%CI)=(3.47 (1.02,11.83)] and weight ≥60kg [Adj. HR (95% CI)=(3.47 (1.02,11.83)]. Conclusion: Substantial magnitude of loss to follow up has been found among patients on ART which significantly affect the overall outcome of HIV/AIDS program of treatment. The independent predictors identified were TB smear positive, male gender, regiment AZT-3TC-NVP and lower weight. So, continuous and comprehensive follow up is necessary to minimize loss to follow up and optimize treatment outcome of people on ART. Mr. Kidane has completed his MSc. from Mekelle University College of health sciences. He is member of different college committees related to quality of education, a postgraduate program coordinator. He has published more than 10 papers in reputed journals.

Biography:

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Abstract Objectives: The aim of this study was to determine antiretroviral drug resistance patterns in patients on chronic HAART presenting with OPC. Methods: An exploratory survey was performed among HIV-infected patients on HAART for minimum of 24 months presenting with OPC in Nairobi, Kenya. Type (pseudomembraneous or erythematous candidiasis, angular cheilitis) and previous episodes of OPC, CD4-cell counts, duration, regimen and adherence on HAART were compared between patients with detectable (>1000copies/ml) and undetectable HIV-RNA levels. Genotypic resistance testing was performed on those with detectable viral loads. Results: Out of (n=45) patients with OPC, (n=28; 62%) had detectable HIV-RNA levels. The (n=28) patients who mostly presented with pseudomembraneuos candidiasis (n=26; p<.0001), had significantly more previous episodes of OPC (55% versus 18%; P<0.0373) lower median CD4 cell counts (74 versus 521; P<.0001) and higher HIV-RNA median plasma levels (111,191 copies/ml versus <20; P<.0001). The sensitivity (0.96) and specificity (0.87) of pseudomebraneous candidiasis to predict virological failure was high. HIV genotyping performed in 22 of the 28 patients showed that most (18/22) had drug resistance mutations of which 12/18 had Lamivudine-associated M184V mutation, 14/18 had TAMS and 16/18 had NNRTI mutations. One patient had major PI mutations. Conclusion: Virological failure and drug resistance mutations including TAMs should be suspected in patients on chronic HAART that present with pseudomembraneous candidiasis. We propose to include recurrent OPC in the WHO clinical criteria for HAART failure as well as to establish clinical training sessions to build competences among health care providers. Trial registration: Netherlands Trial Register NTR2697 (date registered 13th January 2011).

Biography:

Dr. Prabagaran Narayanasamy is a faculty member in the Department of Pathology and Microbiology at the University of Nebraska Medical Center. He received his Ph. D at IIT in Organic Chemistry and did his postdoctoral studies at North Dakota State University, Harvard University and University of Illinois Urbana-champaign. Later, he joined as a Research Scientist at Colorado State University to explore drug discovery. He has been a faculty at University of Nebraska Medical Center since 2011. Dr. Narayanasamy’s research interests are on development, delivering and discovering drug for anti-mycobacterial medicine and antiretroviral therapy. For antibacterial drug discovery - glyoxalase, quorum sensing, MEP and menaquinone pathway are utilized. For antiviral drug discovery NRTI concept is used. Conventional (HIV and TB) drugs and new inhibitors are used in nanoformulation to generate active nanomedicine for sustained drug release through macrophages. In vitro and In vivo characterizations of drug like compounds were also carried out. In addition, metabolites are evaluated in the infected brain for characterizing neurodegenerative disorders. He has funding from NIH and also in study sections.

Abstract:

Human immunodeficiency virus (HIV) infection and Mycobacterium tuberculosis (TB) are responsible for two of major global human infectious diseases. As a result co-infection is common. Thus, improvements in drug access and simplified treatment regimens are needed immediately. One of the key host cells infected by both HIV and TB is the mononuclear phagocyte (macrophage). Therefore, we hypothesized that one way this can be achieved is through nanoformulated drug that ideally would be active against both HIV and TB. We manufactured nanoformulations of antibiotics, antiretroviral therapy and Gallium (Ga). The manufactured nanomedicine was used to study the drug uptake and release kinetics by monocytes (MDM) treated with macrophage colony stimulating factor (MCSF). Finally the efficacy study with nanomedicine was done against Mycobacterium smegmatis, HIV and co-infection in macrophages. The nanoparticle (NP) agent enhanced the drug uptake and maintained the drug release. It also inhibited growth of both HIV and mycobacterium in the macrophage separately and while co-infected for up to 15 days following single drug loading. The NP was also found in all compartments of macrophage in subcellular trafficking study. The multi targeted prolonged-acting NP was effective up to 15 days after single drug loading. The subcellular trafficking of NP was determined and the presence of NP in all the compartments confirmed the multi-targeting approach. These results provide a potential new approach to treat HIV-mycobacterial co-infection that could eventually lead to improved clinical outcomes. Supported by grants from the National Institute of Health

Biography:

Joan Smith-Sonneborn is a Professor Emeritus who completed her PhD at Indiana University, Postdoctoral studies at Brandeis University, and University of California Berkeley. She was working as a Research Associate, University of Wisconsin, Professor at University Wyoming and did sabbatical training at U Southern California Berkeley, and Monash University, Australia. She has 68 articles in reputed journals, over 100 presentations in United States, England, Germany, Monte Carlo, Sicily, Italy, Japan, Malaysia, Canada, Mexico, and Argentina. She is presently a member of NYAS, GSA, and NSCA. She has served as Chair of the International Ciliate Conference, Gordon Conference, and reviewer for grants, journals, and Advisory Board for FDA.

Abstract:

Telomerase can be touted as the miracle anti-aging enzyme that reverses the age-related attrition of telomere ends. But recent studies reveal a darker side of telomerase related to its role independent of telomere functions in cancer, inherited, and infectious disease states. The catalytic subunit of telomerase, TERT telomerase reverse transcriptase, is known to function as an RNA dependent DNA polymerase in the nucleus (TERT-TERC), as an RNA dependent RNA polymerase, anRdRP (TERT-RMRP), in mitochondria, and, as TERT alone interacting with master regulators of cell function, and/or protection of mitochondrial DNA. The mitochondrial TERT-RMRP is the only mammalian RdRP identified to date, capable of generating double stranded RNA, and acting as a substrate for dicers to generate small interfering RNA’s. TERT also shows conservation of the viral close-right-handed polymerase structure, like viral polymerases, and is capable of producing cDNA, double stranded RNA, and small RNA’s capable of controlling mammalian genome expression. TERT also shows promiscuous partnering with RNA elements, RMPR, tRNA, and TERC. TERT, with its versatile viral-like functions, seems like a valuable hostage for use in viral infection. Telomerase is known to increase in retroviral infections, and can increase mitochondrial oxidative stress resistance by inhibition of pro apoptotic genes, a known role of independent of TERT-TERC. Since telomerase is already a target in anti-cancer drugs, the literature provides a valuable reservoir for potential anti-retroviral drugs. Drugs that inhibit the siRNA’s that promote viral survival, or are necessary for viral survival, offer promising potential success in anti-retroviral therapy, such as interfering with the HIV-TAR miRNA, used to establish initial virus survival and replication or allowing release/stimulation of cell immunity processes using hormetic mimetics.

Biography:

Abstract:

Background: Short intra-weekly cycles of anti-HIV combinations have provided intermittent yet effective therapy. The concept is now extended to 94 patients on treatment (Rx) 4 days a week (d/wk) or less over a median 2.7 discontinuous treatment-years per patient. Patients & Combinations: On steadily suppressive combinations, 94 patients volunteered to 5 and 4 d/w treatment, or directly reduced stepwise to 4, 3, 2, 1 d/wk in respectively 94, 84, 66, 12 pts, on various triple (standard) or non-registered quadruple antiviral combinations. Results: 4 d/wk RX aggregated 165 intermittent treatment-years recording no failure over 87 average Rx-weeks per patient; with 63/94 pts successfully passing the 144 weeks bore on any of the antiviral combinations prescribed. On highly (3d/wk) or ultra (2 d/wk) or hyper (1d/wk) intermittent Rx, HIV RNA surged >50 copies 4 weeks apart in 18 instances (6.8 viral escapes / 100 discontinuous maintenance-years), relatable to: Erratic adherence to regimen or follow-up (3 patients); base-drug at ½ the daily recommended dosage (8 pts) ; and/or overlooked archival resistant HIVs from antecedent treatment failures (6pts). Aside from such human blunders, HIV unexpectedly rebounded in 3 and 1 patients respectively on 2 and 1/d/wk Rx, = 2.2 intrinsic viral escapes per 100 ultra-hyper intermittent treatment-years. All 18 escapes were countered by 7 day-a-wk salvage combinations and 11/18 has been back to a second course of intermittent therapy 4 days a week or less. Both cell-activation markers on the surface of T- lymphocytes, and cell-bound HIV DNA levels remained stable or declined. CD4/CD8 ratios rose to ≥1 in 35% of patients, while CD4 counts went ≥500/μl in 75% - from 7% and 40% respectively on unremitting therapy. Conclusion: In our aging, long-HIV enduring, multi-treated patient cohort, ICCARRE cut into current overmedication by 60%, offering the equivalent of 3 drug-free/ 3 virus-free remission years per patient ≈3 million Euros unspent for just 94 patients, at the cost of 2.2 intrinsic viral failures per 100 ultra-hyper intermittent treatment-years. On-treatment 4 days a week would universally provide 40% cuts in dispensable medications at no risk of failure.

Biography:

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The Raf kinase was discovered 30 years ago as a retroviral oncogen. It is activated in several human cancers and a major target of recent drug therapy. However, the Raf kinase can also induce differentiation instead of proliferation, depending on the cell-type and growth factor stimuli. We described a cross-talk between the Raf-kinase and the PI3Kinase pathways which influences the cellular responses. Thus, in cells where the Raf kinase is normally inducing differentiation, anti-cancer drugs may induce prolieration and cause an undesired opposite effect. This has been observed repeatedly in patients treated with one of the novel drugs against the Raf kinase. We also described previously two negative feedback loops inducing upstream signalling from Raf-MEK to the EGF receptor. Again, inhibition of this loop by drugs against Raf may induce the opposite effect. This was indeed observed in patients and therefore recently a dual therapy was applied in order to compensate for the loss of the negative feedback, with some therapeutic success. Considering the unexpected counterintuitive effects of Raf kinase inhibitors and novel therapeutics it is worth discussing the known regulatory mechanisms we have described, and avoid side-effects.

Biography:

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Despite the encouraging development of a preventive vaccine for human papillomavirus (HPV), it cannot improve ongoing infections. Therefore, a new vaccine is urgently needed that can prevent and treat cervical cancer, and cure pre-cancerous lesions. In this study, we constructed two peptide-based vaccines. The first was a short-term, long-peptide (ST-LP) vaccine that simultaneously targeted three key carcinogenic epitopes (E5-E6-E7) on HPV16. We tested this vaccine in murine TC-1 cells infected with a recombinant adeno-associated virus (rAAV) fused with HPV16E5 DNA (rTC-1 cells), which served as a cell model; we also tested it in immune-competent mice loaded with rTC-1 cells, which served as an ectopic tumor model. The ST-LP injections resulted in strong, cell-mediated immunity, capable of attacking and eliminating abnormal antigen-bearing cells. Furthermore, to prolong immunogenic capability, we designed a unique rAAV that encoded the three predicted epitopes for a second, long-term, long-peptide (LT-LP) vaccine. Moreover, we used a new immune strategy of continuous re-injections, where three ST-LP injections were performed at one-week intervals (days 1, 8, 15), then one LT-LP injection was performed on day 120. Our in vitro and in vivo studies revealed that this strategy could boost the immune response to produce longer and stronger protection against target cells, and mice were thoroughly protected from tumor growth. Our results showed that priming the immune system with the ST-LP vaccine, followed by boosting the immune system with the LT-LP vaccine could generate a rapid, robust, durable cytotoxic T-lymphocyte response to HPV16-positive tumors.

Biography:

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cases occur in Africa including Ethiopia. The transmission of HIV from infected mothers to babies could occur during pregnancy, delivery and breastfeeding. For women to take advantage of measures to reduce transmission, they need to know about Mother To Child Transmission (MTCT) of HIV and their HIV status. The aim of this study was to assess the knowledge on MTCT and utilization of services designed for Prevention of Mother-To-Child Transmission (PMTCT) of HIV/AIDS among pregnant women. Methods: Community based cross-sectional study was conducted at Hossana town from March 3-28/ 2014 using pre-tested questionnaire and structured interviews. The collected data were analyzed using SPSS version 16. Descriptive statistics and logistic regression analysis were done. Result: Out of the 417 pregnant women interviewed, 370 (88.7%) responded that they know MTCT of HIV, 377(90.4%) mothers tested for HIV during current pregnancy and 354(93.9%) shared test result to their husband. Few, 23(6.1%) did not disclose test result due to fear of divorce. The main reason for HIV testing, according to this study, is to know self-status. Knowledge of Mother to child transmission was the independent predictor of utilization of the services rendered for PMTCT of HIV/AIDS. Conclusions: More than three-fourth of pregnant women knew about MTCT of HIV. Nine women in every ten tested for HIV during current pregnancy and shared test result to their husband. Knowledge of mother to child transmission of HIV/AIDS was the independent predictor of utilization of PMTCT services. Thus, improving awareness of pregnant women about MTCT of HIV/AIDS and its prevention strategies by means of health care providers in maternal and child health service units should be strengthened.

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Obtaining biological functionalities such as pharmacological activities, disease processes, physiological and structural properties by analyzing the sequence information attributed to them has preliminarily been considered impracticable. This is because there are no known procedures that could be engaged in order to help uncover biological functionalities encoded in the sequence information. It has also been recognized that most biological functionalities of drugs consisting of alkaloids, flavonoids, Terpenes, steroids, etc could be expressed in one gene/protein or the other. For example, Multidrug-resistance transporter gene (MDR1) encoding genes for P-glycoprotein and CYP450, which play vital roles transport and metabolism of Antiretroviral agents have been identified. This MDR1 gene regulates the activity of some antiretroviral drugs. Similarly, anti-bacteria multi-drug resistance genes (MDR1 and MDR2) control the activities of Ciproxfloxacin (an alkaloid), Penicillin (a Beta-lactamase) and tetracycline (a polyketide), Gentamycin (an Aminoglycoside) antibiotics, etc. The same applies to anti-Malaria agents and others. In effect, activities of a wide range of therapeutic agents belonging to numerous groups could be read from one gene/protein they express. Furthermore, genes/proteins encoding therapeutic agents can provide as much information as the therapeutic agents. Direct translations of sequence information into biological functionalities were truly impossible until 1985 when researchers saw proteins/peptides as signals (numerical sequences) instead of piece of fish, meat, bowl of beans or an enzyme. This has opened up a novel area of research. As signals, proteins and peptides were analyzed using techniques that have help develop technologies like Radar, Image Processing and Speech Detector. This technique is called Digital Signal Processing (DSP). This presentation demonstrates how biological functionalities could be translated directly from their sequence information using a DSP technique called Informational Spectrum Method (ISM) and two peptides VIPMFSALS and CAPAGFAIL. This technique has offered direct translation of sequence information into various bio-functionalities. It compared the efficacy of two anti-retroviral agents (Enfurvitude and Sifurvitude) as well as two starter materials (P18 and P32) for the designing of anti-malaria vaccines. It explained the HIV progression to AIDS. It identified the origins of HIV-1 non B subtypes that infected American soldiers on Foreign Service. It elucidated the molecular mechanisms to protection offered to the heart by Influenza vaccines as well as Emilins to Anthrax antigen. It calculated biological functionalities and has helped develop a biomedical device, Computer-Aided Drug Resistance Calculator. Based on the level of biological functionalities uncovered from sequence information using DSP procedures, it can be ascertained that it has now become feasible to obtain same results we get from clinical laboratories by analyzing sequence information involved. Because this approach is rational, it is recommended that it be fully exploited.

Biography:

Abstract:

Long Interspersed Nuclear Elements (LINE-1) and endogenous retroviruses represent the most abundant families of retrotransposable elements in mammalian genomes. They encode a reverse transcriptase (RT) protein as part of the ORF2, which is required for their own mobilization as well as that of non-autonomous Alu/SINE retrotrasposons. We have shown that LINE-1-derived ORF2p, encoding RT, is abundantly expressed and specifically localized in murine embryo early stages and in a variety of human cancers, while being virtually absent from normal somatic differentiated tissues. We also detected a progressive increase of LINE-1 and SINE copy numbers in murine preimplantation developmental stages and in tumor progression. Conversely, RT inhibition arrests early embryo development and inhibits cancer cell proliferation, promotes their differentiation and antagonizes tumor growth in animal models. In line with this, the nonnucleoside RT inhibitor efavirenz recently proved effective in phase II trials with prostate carcinoma metastatic patients. To get insight into the RT mechanism in diverse processes as early embyrogenesis and cell transformation, we have examined global expression profiles in native and RTinhibited cells. We find that: i) RT inhibition causes a global reprogramming of expression of coding genes, microRNAs (miRNAs) and genomic ultra conserved regions (UCRs) (the latter two are enriched in Alu sequences, often organized as pairs of inverted repeats), ii) Alu- and LINE-1-containing RNA:DNA hybrid molecules are abundant in cancer but not in normal cells or in RT inhibitor-treated tumor cells. We therefore propose that the abundant RT in embryos and cancer cells reverse-transcribes RNA precursors, generating RNA:DNA hybrids that affect the overall production of various RNA classes, including regulatory miRNAs, with an ensuing impact on global gene expression. RT inhibition restores the ‘normal’ RNA expression profiles. Thus, LINE1-RT drives a novel regulatory mechanism, required in early embryogenesis, which, when erroneously reactivated in adult life, drives cell transformation and tumorigenesis.

Biography:

Dr Donatien Mavoungou is the Director of the Research Center on Hormonal Pathologies (CRPH) Gabon. Invited Member at McGill University Aids Center (Montreal) and at Genomic Chair of Canada. He authored more than 150 publications. He discovered IMMUNOREXTM DM28 inhibiting HIV-1 replication. He is Professor of Biochemistry and Endocrinology at the University of Health Science Libreville, Gabon. Field: HIV, Metabolic disorders, HAART, Hormonal Therapy. He received the Prize of the National Center for Scientific Research in 2009 and 2012. (Gabon). Blue Ribbon Presenter, recipient of the Glaxo Smith Klein Award at ISHIB, Orlando (USA, 2007). Member of WABT Academy

Abstract:

ImmunorexTM as an antagonist of HIV-1 Tat, removes the overproduction of IL-10, contributing to the restoration of TH1 and TH2 balance and prevents cooperation between HIV-1 and co-infecting pathogens. Mass spectroscopy of ImmunorexTM revealed presence of DHEA, salicin, threonine ,fatty acids among others molecules, having antiviral, antibacterial, anti pyretic, anti inflammatory and anti cancer activities. Objective: analyse ImmunorexTM lipids fraction (LCF) as a putative source of antibiotics. LCF microbicidal activity was assessed by the sensitivity using the methods of Chapman and Sabourreau. Escherichia Coli (E.coli), Staphylococusareus, Pseudomonas and Klebsiella were isolated from the urine of HIV-1-infected patients diagnosed by western Blot technique. Various antibiotic disc (ABD) have been impregnated at 25°C with the LCF (Group 1), then dried and incubated with various germs isolated in parallel with native counterparts in Group 2. The diameters were measured (cm) to compare sensitivity between groups. LCF showed significant increase of respective antibiotics via diameter (cm) : 16→28; 10→18 + Ac Claulamique for staphylococcus areus, LCF increased sensitivities of Amoxicilline, Cefotaxime (0→32), Netelmicine ( 20→34); Rifampicine (13→22).cefotaxime (21→24); Aztreonam (41→48) and Amoxicilline Ac clavulamique ( 0→18) for E. coli while for Klebsiella, sensitivities to ABD was: Amoxiciline + Ac. clavulamique (20→28;24→30); Neteilmicine (20→24; 20→28); LCF showedwhile for Pseudomonas, negligible sensitivities to ABD. ImmunorexTM revealed potential microbicide activity that may contribute to block HIV-1 infection and the cooperation with co-infecting pathogens, protecting the unavoidable TH1 and TH2 balance in a drug and vaccine approach against HIV-1, including Tat Oyi vaccine as suggested by Biosantech, France.

Biography:

Abstract:

The case definition of AIDS encompasses a spectrum of infection and malignancies, labeled as opportunistic infections. In recent years, numerous studies have outlined the emergence of opportunistic gastrointestinal protozoa that have caused diarrheal illness among HIV/AIDS patients thus affecting the quality of life in these patients. These infections are the leading cause of morbidity and mortality among these patients especially in third world countries (with Cameroon inclusive) where assess to antiretroviral is still a major public health problem. The purpose of this study was to determine the prevalence of opportunistic intestinal protozoan as well as correlate the mean parasite density with CD4+ count levels among HIV/AIDS patients attending the North West Regional Hospital Bamenda, Cameroon. A hospital base cross-sectional study design was conducted were 98 stool samples were collected from HIV/AIDS volunteers. The stool samples were concentrated using the modified Ziehl-Neelsen(Zn) technique for the detection of oocyst of the opportunistic protozoans. Also EDTA blood samples were collected for determination of CD4+ count using a FACS count analyzer. Intestinal protozoans were detected in fourteen (14) of the subjects giving a prevalence of 14.2%. Cryptosporidium parvum (12.2%) was the most predominant parasite identified among the study subjects followed by Cyclosopracayetenensis (2.2%). Majority of the participants (56%) had a low CD4+ count and recorded the highest prevalence (11.2%) of intestinal protozoans which was statistically significant (p< 0.05). This study also reported negative correlation (r= -0.66) between mean parasite density and CD4+ among HIV/AIDS subjects.This study thus ascertains the existence of intestinal protozoans among HIV/AIDS subjects attending the North West West Regional Hospital, Bamenda.

Biography:

Abstract:

Integration of retroviral elements into the host genome is a phenomena observed among many classes of retroviruses. Much information concerning integration of retroviral elements has been documented based on in vitro analysis or expression of selectable markers. To identify possible integration events of the LTR retrotransposon, Tf1, within silent regions of the S. pombe genome, we focused on performing an in vivo genome-wide analysis of Tf1 integration events from the nonselective phase of the retrotransposition assay. After analyzing 1000 individual colonies streaked from four independent Tf1 transposed patches under nonselection conditions we detected a population of G418S/neo+ Tf1 integration events that would have been overlooked during the selective phase of the assay. Interestingly, further RNA analysis from the G418S/neo+ cells revealed 50% of clones expressing the neo selectable marker. S. pombe cells have in place mechanisms such as DNA methylation, histone modification, and RNA interference (RNAi), to control retroviral activity. There are increasing reports suggesting that RNAi may play a role in silencing virally infected eukaryotic cells. RNAi was also shown to be involved in the inhibition of viruses and silencing of viruses in plants, insects, fungi, and nematodes. We utilized non denaturing Polyacrylamide gel electrophoresis (PAGE) and Northern Blot hybridization using a DIG-labeled neo probe to detect the presence of microRNAs in G418S/neo+ clones.

Biography:

Donatien Mavoungou is the Director of the Research Center on Hormonal Pathologies (CRPH) Gabon. He is the Invited Member at McGill University Aids Center (Montreal) and at Genomic Chair of Canada. He authored more than 150 publications. He discovered IMMUNOREX TM DM28 inhibiting HIV-1 replication. He is Professor of Biochemistry and Endocrinology at the University of Health Science Libreville, Gabon. Field: HIV, Metabolic disorders, HAART, Hormonal Therapy. He received the Prize of the National Center for Scientific Research in 2009 and 2012. (Gabon). He is the Blue Ribbon Presenter, recipient of the Glaxo Smith Klein Award at ISHIB, Orlando (USA, 2007). He is the Member of WABT Academy.

Abstract:

LCF microbicidal activity was assessed by the sensitivity using the methods of Chapman and Sabourreau. Escherichia coli (E.coli), Staphylococcus aureus, Pseudomonas and Klebsiella were isolated from the urine of HIV-1-infected patients diagnosed by western Blot technique. Various antibiotic disc (ABD) have been impregnated at 25°C with the LCF (Group 1), then dried and incubated with various germs isolated in parallel with native counter parts in Group 2. The diameters were measured (cm) to compare sensitivity between groups. LCF showed significant increase of respective antibiotics via diameter (cm): 16→28; 10→18 + Ac Claulamique for Staphylococcus aureus, LCF increased sensitivities of Amoxicilline, Cefotaxime (0→32), Netelmicine ( 20→34); Rifampicine (13→22).Cefotaxime (21→24); Aztreonam (41→48) and Amoxicilline Ac clavulamique ( 0→18) for E. coli while for Klebsiella, sensitivities to ABD was: Amoxiciline + Ac. clavulamique (20→28;24→30); Neteilmicine (20→24; 20→28); LCF showed while for Pseudomonas, negligible sensitivities to ABD. ImmunorexTM revealed potential microbicide activity that may contribute to block HIV-1 infection and the cooperation with co-infecting pathogens, protecting the unavoidable TH1 and TH2 balance in a drug and vaccine approach against HIV-1, including Tat Oyi vaccine as suggested by Biosantech, France.

Biography:

Dawit Assefa Research student at Clinical Virology, Malmö located at Sweden

Abstract:

Background: In Ethiopia, free access to ART has been expanded rapidly since 2005. With the rapid scale up and decentralization of the ART service, the emergency and transmission of HIV drug resistance (HIVDR) will be a major problem. In view of this, we evaluated the prevalence of transmitted HIVDR (TDR) in recently HIV-1 infected adults in Gondar town, Ethiopia using WHO threshold surveillance method. Methods: A cross-sectional survey was conducted among antiretroviral naive adults who were seeking HIV diagnostic testing in VCT site in Gondar university hospital and Gondar Health center, from August 2011 to December 2012 using the WHO recommended eligibility criterion. A total of 84 study participants fulfilling the inclusion criterion were consecutively enrolled and blood specimen were collected. HIVDR genotyping was done using the in-house assay. Sequences were interpreted using the calibrated population resistance (CPR) tool of the Stanford University database according to the 2009 WHO surveillance drug resistance mutation (SDRM) list. Result: Among the 84 participants, 70 (83.3%) were female and mean age was 21years (range: 18–24 years). Amplification and sequencing was successful for 67(79.8%) of specimens. Using the WHO recommended truncated sequential sampling technique, among the first 47 sequenced specimens, 3 were found to harbor major HIVDR mutation associated to non-nucleoside reverse transcriptase inhibitor (NNRTI), suggesting moderate prevalence (5%–15%) of TDR in the study area. The mutation detected were K103N (n=2) and G190S (n=1). Including all sequenced sample in the analysis (n=67), 4(6%) were found to have major HIVDR mutations (K103N (n=2), G190S (n=1), and Y181Y (n=1)) associated to NNRTI. However, no nucleoside/tide reverse transcriptase inhibitor (NRTI) or protease inhibitor (PI) associated major HIVDR mutation were detected. Conclusion: In comparison to previous studies done in the Gondar town, our result showed an increase in prevalence of TDR, which could be associated to the ART scale up. As the TDR HIV-1 may seriously affect the efficacy of first line ART, the moderate levels of TDR observed in this study area indicate the need for continuous surveillance of TDR in Gondar town and in different region of Ethiopia to optimize treatment efficacy of the current ART and improve the drug resistance strategy within a country.

Biography:

Sergei Nekhai has a Ph.D. degree in biophysics from St. Petersburg Nuclear Physics Institute, Russia. He did his postdoctoral training at the George Washington University. He serves as co-Director for Howard University’s Center for Sickle Cell Disease and Director of Proteomics. He studies HIV-1 transcription, pulmonary hypertension in sickle cell disease and iron metabolism with the focus on ferroportin mutations that might pose a risk for iron overload and worsen HIV-1 infection in African-Americans. He has more than 50 publications in basic virology and hematology journals and serves as a PI and co-PI on several NIH-funded grants.

Abstract:

HIV-1 transcription is activated by Tat protein that recruits CDK9/cyclin T1 to TAR RNA. We previously showed that Tat also binds to protein phosphatase-1 (PP1) through the Q35VCF38 sequence and translocates PP1 to the nucleus. PP1 dephosphorylates CDK9 and activates HIV-1 transcription. We recently identified PP1-targeting small molecule, 1H4, that prevented HIV-1 Tat interaction with PP1 and inhibited HIV-1 gene transcription. Using the model of 1H4-PP1 complex we iteratively designed and synthesized follow-up libraries that were analyzed for the inhibition of HIV-1 transcription and toxicity. We obtained a tetrahydroquinoline derivative, 1E7, which inhibited phosphatase activity of PP1 and also disrupted the interaction of Tat with PP1. We further optimized 1E7 and obtained compound 1E7-03 that inhibited HIV-1 with low IC50, showed no toxicity when administered in mice. The 1E7-03 was also active in HIV-1 transgenic mice preventing death from acute lung inflammation induced by LPS injection. The LPS administration led to neutrophil and macrophages recruitment to the lungs where HIV-1 expression in the lung macrophages prevented neutrophil clearance. Injection of 1E7-03 reduced both macrophages and neutrophil accumulation in the lungs likely due to the reduction of HIV-1 expression. We further analyzed stability of 1E7-03 compound and identified its major metabolites. We also developed 1E7-03 analogs that had improved stability and showed similar activity to the parental compound. Our study shows that PP1 can serve as a target for development of novel therapeutics against HIV-1 to target HIV-1 expression in lungs and potentially other organs. Acknowledgements: This work was supported by NIH Research Grants (1P50HL118006, 1R01HL125005, U19AI109664 and 5G12MD007597), and District of Columbia Developmental Center for AIDS Research grant (P30AI087714). This study was also funded in part with NIH Grant UL1TR000101 from the National Center for Advancing Translational Sciences (NCATS), National Institutes of Health (NIH), through the Clinical and Translational Science Awards Program (CTSA), a trademark of DHHS, part of the Roadmap Initiative, “Re-Engineering the Clinical Research Enterprise.” The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Biography:

Binhua Ling of Tulane University, New Orleans with expertise in Clinical Immunology, Infectious Diseases, Allergology , HIV, AIDS

Abstract:

HIV infected long term non-progressors (LTNP) and elite controllers (EC) maintain enduring control of HIV infection without anti-retroviral therapy (ART). Emerging evidence suggests that cells in tissues such as the gut continue to produce low levels of HIV, low level ongoing viral production occurs in the body even with concurrent ART. Whether LTNP/ EC receiving ART can completely suppress low level ongoing viral replication is unknown. We have previously found that SIV infection in Chinese rhesus macaque (ChRM) is the closest animal model that mimics HIV infection. We have also shown that gut mucosa is a major reservoir in LTNP/EC of SIV infected ChRM. Here we studied SIV mutation and evolution in the gut associated lymphoid tissue to determine whether ART can fully suppress viral replication in LTNP/EC of SIV infected ChRM. Ten animals were treated with reverse integrase inhibitors (tenofovir and Emtricitabine) daily for up to 24 weeks. PBMCs and lymphocytes were isolated from blood, spleen, lymph nodes and gut tissues of 6 sites (duodenum, jejunum, ileum, cecum, colon and rectum) by the end of ART. Nested PCR and single genome amplification (SGA) was performed. SIV envelope gp120 gene from positive samples was amplified. Sequences from PBMC before and after ART were compared with sequences isolated from the 6 sites of the small and large intestine. Phylogenetic trees were constructed. Viral mutation and evolution were analyzed. The size of reservoirs in PBMC and lymph nodes was evaluated by quantitative co-culture assays. We found that plasma viral loads (pVL) in all animals were suppressed and sustained below 28 copies per ml (limitation of detection) during ART. Two animals had viral rebound after approximately 2 weeks of cessation of ART. Viral DNA loads were detectable in either PBMCs and/or lymphocytes from different gut sections at variant levels by the end of ART. The frequency of detective PCR positives was higher in large intestine than small intestine, indicating large intestine is a major reservoir. A total of 245 sequences were analyzed. While envelope sequences showed limited viral mutations in some animals, viral evolution was observed in different sections of the gut in other animals even received 24 weeks of ART with sustained undetectable pVL. The size of viral reservoirs in blood reduced in LTNPs/ECs after ART. However, spleen and lymph nodes had higher resting CD4+ T cells carrying replication competent SIV than blood by the end of ART. These data suggest that low level of ongoing viral replication may still occur in some LTNP on ART. The SIV/ChRM non-human primate model is excellent for testing novel therapeutic interventions on typical progressors and LTNP/EC for HIV eradication.

Biography:

Tao Lu is a tenure-track Assistant Professor and Principle Investigator at Department of Pharmacology and Toxicology and Department of Biochemistry and Molecular Biology, as well as a member of Experimental and Developmental Therapeutics Research Program at Simon Cancer Center at Indiana University School of Medicine. She obtained her Ph.D. degree from University of Toledo, School of Medicine. She then did her postdoctoral training with the world renowned scientist Dr. George R. Stark at Cleveland Clinic, Ohio. She has been working on discovery of novel regulators of NFκB, and is particular interested in epigenetic regulation of transcription factors and their role in cancer. She has won several international awards, including the First Place Prize of Young Investigator Award at Tri-Society [Society of Leukocyte Biology (SLB) & International Cytokine Society (ICS) & International Society of Interferon and Cytokine Research (ISICR)] conference and Seymour and Vivian Milstein Young Investigator Award at ISICR & ICS International conference.

Abstract:

Colon cancer is the second leading cause of cancer related deaths in the United States. The nuclear factor κB (NF-κB) is an important family of transcription factors whose aberrant activation has been found in many types of cancer, including colon cancer. Therefore, understanding the regulation of NF-κB is of ultimate importance for cancer therapy. The purpose of this study is to use a novel validation-based insertional mutagenesis (VBIM) strategy to identify novel regulators of NF-κB, and further evaluate their roles in the regulation of NF-κB signaling in colon cancer cells. We infected Z cells (293 derived cells with hyper active NF-κB activity) with VBIM virus to cause the over expression of negative regulators of NF-κB, and then further selected the mutant cells with low NF-κB activity under ganciclovir (GCV) treatment. Targeted gene was then identified by using VBIM specific primers. In a preliminary screen, we identified the novel adenomatous polyposis coli like protein (ALP) gene as a negative regulator of NF-κB. Over expression of ALP led to decreased NF-κB activity by κB reporter assay, while knocking it down had the opposite effect. Furthermore, we found that over expression of ALP in HT29 colon cancer cells greatly reduced both the number and the size of colonies that were formed in a soft agar assay, while using shRNA resulted in an opposite effect, confirming that ALP is a tumor suppressor in HT29 cells. Future experiments aim to further assess the role of ALP in colon tumor formation in a mouse xenograft model. In summary, by using the novel VBIM technique, we identified ALP as a novel negative regulator of NF-κB. This discovery could lead to the establishment of ALP as a potential biomarker and therapeutic target in colon cancer.

Biography:

Corrado Spadafora's laboratory originally discovered that mature sperm cells from a variety of species share the ability to spontaneously take up exogenous DNA molecules and deliver them to oocytes at fertilization: they called that phenomenon cell-mediated sperm-mediated gene transfer (SMGT). That feature was subsequently exploited, in theirs and other laboratories, to generate genetically modified animals in different species.

Abstract:

Long Interspersed Nuclear Elements (LINE-1) and endogenous retroviruses represent the most abundant families of retrotransposable elements in mammalian genomes. They encode a reverse transcriptase (RT) protein as part of the ORF2, which is required for their own mobilization as well as that of non-autonomous Alu/SINE retrotransposons. We have shown that LINE-1-derived ORF2p, encoding RT, is abundantly expressed and specifically localized in murine embryo early stages and in a variety of human cancers, while being virtually absent from normal somatic differentiated tissues. We also detected a progressive increase of LINE-1 and SINE copy numbers in murine preimplantation developmental stages and in tumor progression. Conversely, RT inhibition arrests early embryo development and inhibits cancer cell proliferation, promotes their differentiation and antagonizes tumor growth in animal models. In line with this, the nonnucleoside RT inhibitor efavirenz recently proved effective in phase II trials with prostate carcinoma metastatic patients. To get insight into the RT mechanism in diverse processes as early embyrogenesis and cell transformation, we have examined global expression profiles in native and RT inhibited cells. We find that: i) RT inhibition causes a global reprogramming of expression of coding genes, microRNAs (miRNAs) and genomic ultra-conserved regions (UCRs) (the latter two are enriched in Alu sequences, often organized as pairs of inverted repeats), ii) Alu- and LINE-1-containing RNA:DNA hybrid molecules are abundant in cancer but not in normal cells or in RT inhibitor-treated tumor cells. We therefore propose that the abundant RT in embryos and cancer cells reversetranscribes RNA precursors, generating RNA:DNA hybrids that affect the overall production of various RNA classes, including regulatory miRNAs, with an ensuing impact on global gene expression. RT inhibition restores the ‘normal’ RNA expression profiles. Thus, LINE1-RT drives a novel regulatory mechanism, required in early embryogenesis, which, when erroneously reactivated in adult life, drives cell transformation and tumorigenesis.

Biography:

Lawrence Kleiman completed his Ph D at Johns Hopkins University, and after Postdoctoral studies at the Beatson Cancer Institute in Glasgow, Scotland, he has spent the remaining of his career at the Lady Davis Institute for Medical Research, Jewish General Hospital, in Montreal, Quebec, where he is also a Professor in the Department of Medicine at McGill University. He has published over 140 papers, and has worked for the last 20 years studying HIV-1 replication at the McGill AIDS Centre.

Abstract:

During HIV-1 infection, the conversion of the HIV-1 RNA genome into a double-stranded DNA that can be integrated into the host cell’s genome is accomplished by reverse transcription. The initiation of this reverse transcription requires a specific cellular tRNALys3 to act as a primer. This tRNA is annealed during assembly of new HIV-1, i.e., the infecting virus already contains annealed tRNALys3. The 3´ 18nts of tRNALys3 anneal to a complementary 18nt sequences in the viral RNA termed the primer binding site (PBS), and in this presentation, we will discuss the cellular and viral factors that promote the ability of the tRNALys3 to locate the 18nt sequence within the >9 KB comprising the HIV-1 viral RNA genome. This is associated with the formation of an early HIV-1 assembly intermediate at the site of the Gag/GagPol translation, whose components include Gag, GagPol, lysyl-tRNA synthetase (LysRS), and tRNALys3. This intermediate contains an increased concentration of tRNALys3due to a specific interaction between viral Gag and LysRS, and is able to bind through LysRS to a viral RNA structure near the PBS that resembles tRNALys3. This initial cytoplasmic annealing of tRNALys3 to viral RNA is part of a multi-staged annealing process, and not only functions in promoting reverse transcription, but also facilitates a conformation change in the 5´-untranslated region of viral RNA that promotes viral RNA dimerization.

Biography:

Jaffer shiffa, is a professor and Consultant Internist of Jimma University which is located in Ethiopia

Abstract:

Background: In Ethiopia where there is no strong surveillance system and diagnostic facilities are limited, the real burden of Tuberculosis (TB) lymphadenitis is not well known. Therefore, we conducted a study to estimate the prevalence of TB lymphadenitis in Southwest Ethiopia. Methods: A community based cross-sectional study was conducted from February to March 2009 in the Gilgel Gibe field research area. A total of 30,040 individuals 15 years or older in 10,882 households were screened for TB lymphadenitis. Any individual 15 years or older with lumps in the neck, armpits or groin up on interview were considered TB lymphadenitis suspect. The diagnosis of TB lymphadenitis was established when acid fast bacilli (AFB) smear microscopy of fine needle aspiration (FNA) sample, culture or cytology suggested TB. HIV counseling and testing was offered to all TB lymphadenitis suspects. Descriptive and bivariate analysis was done using SPSS version 15. Results: Complete data were available for 27,597 individuals. A total of 87 TB lymphadenitis suspects were identified. Most of the TB lymphadenitis suspects were females (72.4%). Sixteen cases of TB lymphadenitis were confirmed. The prevalence of TB lymphadenitis was thus 58.0 per 100,000 people (16/27,597) (95% CI 35.7-94.2). Individuals who had a contact history with chronic coughers (OR 5.58, 95% CI 1.23-25.43) were more likely to have TB lymphadenitis. Lymph nodes with caseous FNA were more likely to be positive for TB lymphadenitis (OR 5.46, 95% CI 1.69-17.61). Conclusion: The prevalence of TB lymphadenitis in Gilgel Gibe is similar with the WHO estimates for Ethiopia. Screening of TB lymphadenitis particularly for family members who have contact with chronic coughers is recommended. Health extension workers could be trained to screen and refer TB lymphadenitis suspects using simple methods.

Biography:

Li Tang, PhD, joined the staff of Roswell Park Cancer Institute (RPCI) as a Research Assistant Professor in 2010 in the Department of Cancer Prevention and Control within the Division of Cancer Prevention and Population Sciences. She earned her doctoral degree in Cancer Prevention and Pathology from the University at Buffalo in 2006; and completed postdoctoral training in Epidemiology at RPCI.

Abstract:

Human endogenous retroviruses (HERVs) are remnants of ancient virus infection. The majority of them are disabled due to mutation and/or deletion. However, HERVK113 and K115 have been shown to have full-length insertion in human genome and retain the ability to encode functional virus proteins in some individuals. Considering the potential role of HERVs in carcinogenesis and a high genetic homology between HERV K and mouse mammary tumor virus, we examined the distribution of HERV K113 and K115 in African American (AA) and European American (EA) women, and investigated their association with breast cancer risk. PCR followed by fragment analysis was used for insertional polymorphism assay. For each HERV, three PCRs were performed to determine whether the insertion is partial or full length. Indeed, a subset of individuals showed insertion of long terminal repeat (LTR) of HERV, instead of whole virus insertion. For both K113 and K115, with or without including LTR insertion, the distribution was significantly different between AA and EA women. A two-fold higher prevalence of HERVs was observed in AA women, showing 51.5% of individuals with at least one copy of either K113 or K115 compared to 23.3% in EA women among controls. The prevalence of HERVs was inversely correlated with proportions of European ancestry. HERV K113 insertion was significantly associated with the reduced risk of breast cancer in both AA and EA women. Near 50% reduction of breast cancer risk was observed in AA and EA women with homozygous insertion of K113 (combined OR, 0.45; 95% CI, 0.25-0.80), while K113 is only HERV identified so far with a capacity to produce virus particles in cells. Interestingly, hormone exposure seems modulate the association of HERV K113 insertion with breast cancer risk. Anti- HERV-K specific antibody has been shown to inhibit breast cancer growth in animal models. Polymorphic insertion of HERV K113 may interact with host immune system and offer some protection against breast cancer.

Biography:

Olabowale Adefolake Omolade is working as a professor in Olabisi Onabanjo University which is located in Nigeria

Abstract:

Background: Hepatitis is an inflammatory condition of the liver and viral hepatitis is a conventional term used to denote hepatitis caused by hepatotrophic viruses (Hepatitis A-G). High prevalence of these viruses was reported in Nigeria. Hepatitis B and C may cause liver cirrhosis and they can be contacted through contaminated blood and blood products. Many blood banks in Nigeria screen for hepatitis B and C using immune-chromatographic screening method (Rapid test strip). This is because these strips are readily available in the market, cheap, requires no electricity for storage, special training or equipment before use. The intent of our study is to compare the sensitivity of this method using an advanced immunological method. Method: 660 potential donors are tested for hepatitis B surface antigen (HBs Ag) and hepatitis C virus antibody using immunechromatographic test strip and ELISA methods. Result: We found out that 38 (5.7%) out of 660 subjects tested positive for HBS Ag using immunochromographic method while 71 (10.8%) were positive using ELISA. None were positive for hepatitis C antibody using immunochromatographic method while 4 (0.6%) subjects were positive using ELISA method. Conclusion: Immunochromatographic method is not good enough to screen blood donors for hepatitis B and C.

Biography:

David Harrich working as a researcher at QIMR Berghofer Medical Research Institute, he belongs to Australia. Associate Professor David Harrich has found a promising way of stopping HIV from causing AIDS. The human immunodeficiency virus (HIV) is currently treated with a cocktail of drugs to stop the virus from causing acquired immunodeficiency syndrome (AIDS). But there is no cure or vaccine

Abstract:

Reverse transcription, the process which converts viral genomic RNA into a double strand DNA, is the central defining feature of HIV-1 replication and a major target for anti-retroviral therapy. Along with reverse transcriptase (RT), viral protein including IN, CA, and Vpr and cellular proteins including eukaryotic translation elongation factor 1A (eEF1A) are important for the process. We previously showed that eEF1A stabilized the RTC in cells and was important for late steps of reverse transcription. Our recent experiments show a direct interaction between RT and eEF1A that can be down regulated by amino acid substitutions in the RT thumb domain, and which lead to downregulated late reverse transcription. Moreover we show that drugs which bind to eEF1A are potent inhibitors of reverse transcription. Experiments to determine if eEF1A binding drugs negatively affect the RTC in cells will be presented. Recently we also showed that a Tat mutant called Nullbasic inhibits reverse transcription. Our recent data shows that Nullbasic is an RT binding protein that is found in viral particles. In vitro uncoating assays show that virions containing Nullbasic undergo accelerated uncoating kinetics, and analysis of cells infected with HIV-1 containing Nullbasic indicates that the levels of RTCs are reduced, consistent with the uncoating defect. We have recently made Jurkat cell line expressing Nullbasic which appear to be highly resistant to HIV-1 infection. In chronically infected Jurkat cells, introduction of Nullbasic can decrease HIV-1 mRNA levels from 150-to 800 fold. The mechanisms responsible for strong inhibition of HIV-1 will be presented. The combined evidence indicates investigations of interaction between RT and viral and cellular proteins could enable new antiviral strategies.

Biography:

Harold Smith is a tenured professor of biochemistry and biophysics at theUniversity of Rochester, School of Medicine and Dentistry. Dr. Smith¹sprimary function at the University is basic research and in this contexthe is fully engaged in biomedical laboratory research as well as trainingpostdocs, graduate and undergraduate students. The Smith lab¹s primaryinterest is understanding the composition, regulation and structure ofmacromolecular complexes involved in regulating gene expression at thelevel of messenger RNA expression and processing. Our focus is on aplatform of enzymes that change the genetic code at the DNA or RNA levelby deaminating cytidine to form uridine. Current data suggest that thisfamily of cytidine deaminase function with other proteins (auxiliaryproteins) as holoenzymes complexes which we refer to as editosomes (forRNA) or mutasomes (for DNA). RNA editing or DNA mutational activity bythese enzymes affect the protein coding capacity of mRNAs and thereby candiversify the proteins that are expressed by cells (the proteome). In 2003he founded OyaGen, Inc as a drug discovery and drug development biotechfocused on anti retroviral therapeutics.

Abstract:

HIV Viral infectivity factor (Vif) evokes the destruction of the host restriction factor known as APOBEC3G (A3G).Vif dimerization has been shown to be essential for Vif binding to A3G and A3G degradation. Experimental data show that in the absence of Vif, virion-assembled A3G will hypermutateproviral DNA during reverse transcription. The open questions are: can Vif be successfully drugged and by sparing A3G, will hypermutation activity be sufficient to inhibit viral replication? High throughput screening (HTS) assays were developed using FRET for Vif dimerization. The screens were used to select compounds with dose-dependent signals by preventing Vif FRET. A secondary assay for Vif-dependent A3G degradation was used to identify compoundsthat preserved A3G. These were triaged for the ability to inhibit pseudotyped HIV replication and to have low cytoxicity. Ion torrent sequencing of integrated viral genomes revealed extensive hypermutation characteristic of A3G preferences. Following medicinal chemistry, a lead was tested in a seven day spreading infection in PBMC. TheVif dimerization FRET assay provided a robust HTS method applicable to a large library of drug-like molecules. Quantitative HTS followed by orthogonal secondary screen and cytotox counter screening enabled selection of limited number of chemistries for pseudo typed viral testing. A lead (SMVDA) was selected with nanomolar IC50. By impairing Vif dimerization, A3G degradation was reduced and viral particle incorporation of A3G was enhanced. Proviral DNA isolated form target cells showed numerous tracts of dG to dA hypermutation that corresponded to multiple nonsense codon and sense changes. 17 different clades and 8 drug resistant strains of HIV infecting PBMC were sterilized by seven days following a single dose of SMVDA. Conclusions: Drugging Vif led to massivedG to dAhypermutation of HIV proviral DNA, such that the protein coding capacity of the virus would be severely compromised. Inhibitors of Vifare broadly neutralizing and inhibited all drug resistant strains of HIV tested. DruggingVif therefore is achievable and the data suggest that this will serve as a firewall for viremia induced by activating reservoirs as well as a solution for rescue and salvage therapies.

Biography:

Hone Anagaw is working in Jimma University, which is located in Ethiopia

Abstract:

Background: Client satisfaction and adherence on ART service were an important task for care providers to increase service utilization and to respond to HIV emergency; however, Other provider-defined criteria is far from ideal if as a result of the service that the patient is unhappy or dissatisfied. There is, then, a sound rationale for making the organization and delivery of health service responsive to consumer opinion. Objective: The aim of this study is to assess client satisfaction and adherence on an ART service provision in Jimma University specialized hospital. Methods: A cross sectional study involving both quantitative and qualitative data collection methods was conducted from May 1-30, 2010. A total of 337 Adult PLWHA on ART for at least 3 months were the study participants. Systematic sampling technique was used to select the study subjects. Data were collected using structure questionnaire, check lists and semi structure interview guide. After clearing and checking for consistency data were coded, entered and univariate and multivariate analysis was carried out using SPSS version 16.0. qualitative data‘s were transcribed and narrated under themes. Result: A total response rate of 100% from 337 sample size was obtained. Among those 203(60.2%) were females. Two hundred thirty one (68.5%) of respondents scores ≥ mean which means 68.5% satisfied relationship with their care providers. In this study, in which adherence was measured using a self report method, 95.5% of patients were adherent with ≥95% prescribed doses. Marital status, occupation, and waiting time was found to be associated significantly with adherence [OR-136, 95% CI 019 - 0.997], [OR 9.341, 95%CI 1.189-73.358], [OR 9.88E-24, 95% CI 1.759E-24-5.550E-23] respectively. Conclusion and Recommendation: The result of the study showed that assessment of client satisfaction and adherence in Jimma University specialized hospital is high adherence rate inspite of satisfaction. Overall client satisfaction and patient, provider relationship satisfaction rate were low. However measured by self report method, adherence to ARV treatment in this study was seems to be encouraging. Working with other religious leaders, and community leaders to strengthen adherence status are recommended.

Biography:

Dr. Sando worked for 5 years as the Monitoring & Evaluation Team Lead at the Ministry of Health and Social Welfare in the Epidemiology Unit of the National AIDS Control Programme (NACP). Prior to his work at NACP, Dr. Sando was a Medical Officer In-Charge at the Tanzania Heart Institute (THI), supervising and overseeing daily medical activities at the facility. Dr. Sando has extensive research experience, mostly pertaining to HIV/AIDS in Tanzania. Dr. Sando received his Doctor of Medicine at Muhimbili University College of Health Sciences (MUCHS), a Master’s of Science in Health Monitoring and Evaluation at Jimma University in Ethiopia, and a Master’s of Science in Epidemiology from Harvard School of Public Health..

Abstract:

Background: The impact of expanded access to HAART on the epidemiology of patients enrolled in HIV care and treatment services is poorly documented. In this study we describe the temporal trends in; a) baseline characteristics b) 12-months lost to follow up c) mortality of patients in 28 public facilities in Dar es Salaam, Tanzania. Methods: Epidemiological, clinical and immunological data of adult patients aged 15 years and above, enrolled October 2004 to September 2012 were collected at baseline and 12 months. Year of enrollment was treated as independent variable and binomial logistic regression model was used for trend tests. P-values under 0.05 were considered significant. Results: A total of 109943 adults with median age of 35 years were enrolled during the study, 71% were female. There was increased mean baseline CD4 (163 to 227 cells/uL, `P<0.01), decreased proportions with advanced disease stages III or IV (72% to 67%, P<0.01), decrease percent’s diagnosed with PCP (P=0.04). We found no significant differences in; mean cholesterol level (P=0.08), systolic blood pressure (P=0.3), esophageal candidiasis (P=0.052), diagnosed with TB (P=0.13) and for women those who are pregnant during first visit to the clinic (P=0.8). There was an increase in mean Hemoglobin level (9.9 to 10.5g/ dL, P=0.02), it was found that over time there was a decrease in enrollment in hospital level facilities and increase in health centers and dispensaries (P<0.001). Increase in patients who are lost to follow up in the first 12-months, 8.9% (2004/05) to 18% (2009) which started to decrease in following years (P < .0001). We also found decrease in both patients who are reported dead within 30 and 90 days after enrollment (P<0.01). There was overall increase in 12-months mortality rate from 8.8%(2004/05) to 21% (2008), which was followed by decrease from 15% (2009) to 3.7% in 2012 (P< .0001). Conclusion: Between 2004 and 2012 the expansion of HAART services has demonstrated positive impact both in early enrollment, retention and survival of PLHIV. The findings highlight the need to escalate efforts to improve access to HIV diagnostic testing and HAART.

Biography:

He obtained Ph. D. in France, then he went to USA in 1990 for a first post-doc in the department of biophysics and biochemistry at Oregon State University, followed by a second post-doc in 1992 in the department of chemistry at the University of California, Davis. He was recruited as full senior scientist in 1992 in CNRS (Centre National de la Recherche Scientifique), which is the main French national agency for scientific research. Erwann Loret obtained the GlaxoSmith Kline Drug Discovery and Development Award from an American scientific committee for his contribution to HIV research, the Medal of Honor from the Aix Marseille University and the French National Institute for Intellectual Properties Award (Institut National de la Propriété Industrielle). He is the director of a CNRS lab entitled “Etude de Tat et Recherches Appliquées sur le VIH-1” (ETRAV) located in the faculty of Pharmacy at Marseille. ETRAV is the only lab working on AIDS in Aix Marseille University. He is member of the Editorial Board of scientific journals. He is author and co author of 55 scientific publications referenced in PUBMED (two are in press), most of them having a high impact factor (PNAS, EMBO, JBC, RETROVIROLOGY etc …) and five international patents (PCT) on active compounds against the HIV-1 Tat. His main field of interest is related to the HIV-1 Tat proteins and he is the coordinator of a Phase I/II clinical trial of a vaccine targeting Tat at Marseille that began in 2013.

Abstract:

The human immunodeficiency virus type 1 (HIV) eradication will require elimination of HIV infected cells. No antiretroviral treatments (ART) or vaccine approaches have been able to reduce significantly the level of HIV infected cells in peripheral blood. This inefficacy is generally explained by the presence of a major reservoir of latent HIV infected cells in the central nervous system (CNS) that would be a sanctuary where Cytotoxic T Lymphocytes (CTL) have no access and would refresh peripheral blood with activated HIV infected cells. In this review, the presence of a major reservoir in the CNS appears to be inconsistent with recent clinical studies measuring HIV DNA. The major reservoirs are gut tissue, rectal tissue and the peripheral blood where HIV infected cells survive in an environment containing CTL. Extracellular Tat might protect HIV infected cells from CTL due to its capacity to cross CTL membranes and trigger apoptosis. Evidences of Tat secretion from HIV infected cells are shown with the detection of Tat antibodies in different clinical studies. Presence of neutralizing Tat antibodies in cohorts of patients who were exposed to HIV but who are now seronegative is described. The conclusion of this review is that a vaccine eliciting neutralizing antibodies against Tat might significantly reduce the level of HIV infected cells, what ART or other vaccine approaches have been unable to achieve now. It could be a first step towards HIV eradication.

Biography:

Dr. Sonia Escaich, Ph.D., served as the Chief Scientific Officer at Mutabilis S.A. She is the Member of the Executive Board at Mutabilis S.A. Her initial appointment involved heading up the virology unit in the Sandoz/Systemix Inc. joint venture, working on anti-HIV gene therapy. She later managed various laboratory units in the anti-infective research department at Rhone-Poulenc /Aventis in France; first in the field of antivirals then in the antibacterial department, where she setup and develop new antibacterial projects, including exploratory projects based on virulence inhibition. Dr. Escaich serves as a Director of Mutabilis S.A. She holds a PhD in Microbiology with a specialization in Virology, and a post doctorate in Sandoz in Vienna.

Abstract:

Background: Tat protein is an HIV virulence factor involved in several physiopathological pathways, and shown to be expressed even during HAART. Several studies have shown that an immune response to HIV-1 Tat is correlated with the control of HIV replication in animal models and a slower disease progression in human. Tat-based vaccine candidates were assessed in clinical trials with variable results. Our rational for using Tat Oyi as a novel vaccine candidate is provided by the isolation of the HIV-1 Oyi defective clone in an asymtomatic Gabonese. The Tat Oyi mutant is defective for transactivation. Further more, antibodies generated against Tat Oyi were able to cross-recognize all Tat variants of the five main world HIV-1 clades. Methods: The Tat Oyi-based vaccine candidate was designed using the full length Tat. The immune response to Tat Oyi has been characterized in preclinical models. A clinical trial phase I/II has been started for a therapeutic Tat Oyi vaccine application: In a double blind study, forty eight HIV seropositive were divided in four groups (twelve patients per group) received three consecutive injections of a candidate vaccine composed of 0 (group 1 control placebo), 10 (group 2), 33 (group 3) and 100 μg (group 4) of synthetic Tat Oyi. In each case, the three injections were intradermal and achieved monthly (i.e. M0, M1 and M2). Results: Preclinical studies have demonstrated the potency of Tat Oyi vaccination to generated neutralizing antibodies against conformational epitopes not found in other Tat vaccine candidates. Further more, a vaccination assay with Tat Oyi in a macaque model showed that injected animals had lower viremia levels after heterologous mucosal challenge with SHIV, moreover the virus-infected reservoir cells disappeared with time in the vaccine group. In clinical Phase I trial, the vaccine candidate with Tat Oyi proved to be safe and well tolerated. Conclusions: The specific feature of this vaccine candidate versus other Tat based vaccine candidates relies on Tat 3D conformation and the ability to induce a cross reactive immune response to Tat variants, active against this extracellular HIV target. Biosantech’s vaccine candidate therapeutic effect will be analyzed by measuring the immune response and HIV replication after stopping HAART for 2 months in the ongoing Phase IIa trial.

Biography:

Arangassery Rosemary Bastian was born in Kerala in India, before moving to Kenya with her family at the age of three. In 2006, Arangassery enrolled to study Biomedical Engineering at the Manipal Institute of Technology in India, doing her final year in a partnership programme with Drexel University in Philadelphia, where she graduated with a Bachelor of Science degree in 2009. She is currently studying for there for PhD.

Abstract:

Introduction: Initial entry of HIV-1 into host cells remains a compelling and yet elusive target for developing agents to prevent infection. This step is mediated by a sequence of interactions of a trimeric gp120/gp41 envelope (Env) protein complex with host cells, including initial gp120 encounter with the cellular receptor CD4 and a chemokine co-receptor usually either CCR5 or CXCR4. A peptide triazole class of entry inhibitor leads has been shown to bind to gp120 with close to nanomolar affinity, to suppress protein ligand interactions of the Env protein at both its CD4 and co-receptor binding sites and to inhibit cell infection by a broad range of virus subtypes. Further we have also shown that gold nanoparticle conjugated peptide triazoles lead to 20 fold enhanced potency of their anti-viral effects against HIV-1. Previously bowman et al. has shown that multivalent display of HIV inhibitors on gold nanoparticles (AuNPs) has lead to a substantial amount of poency enhancement. This study will lead to the study of size dependency and density dependency on the gold nanoparticle peptide triazole conjugates to further understanding of their mechanism of action leading to enhanced potency. Materials and Methods: The AuNPs were synthesized using a modified citrate reduction method to obtain size-controlled, stable and monodisperse AuNPs. The peptide (KR13) was conjugated to the AuNP using a direct gold-thiol covalent link. The size and extent of polydispersity of the AuNP-KR13 conjugates were measured using Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS). The HIV-1 viral entry inhibition potencies of KR13 and AuNP-KR13 conjugates were compared using a single-round pseudoviral infection luciferase reporter assay using lab synthesized pseudoviruses. We subsequently tested the effects of KR13 and AuNP-KR13 on the virus particle itself by measuring release of the nucleocapsid protein p24 using ELISA analysis of cell-free virion inhibition. Results and Discussion: Compared to peptide triazole alone, the 20 nm AuNP-KR13 conjugate exhibited a close to 20-orders of magnitude enhancement of infection inhibition activity (Table) and further with increasing size of AuNP, the potency was enhanced further with the 120 nm AuNP-KR13 having nearly 1600 fold enhancement (Table). KR13 and AuNP-KR13 conjugates are specific for HIV-1 envelope. There was no significant in vitro cytotoxicity observed for either KR13 or AuNPKR13 conjugates. Further the residual virion that was generated upon treatment with KR13 was immunoreactive while the residual virion generated upon AuNP-KR13 treatment was completely inactive. The mechanism was studied by observing the morphological changes caused by both KR13 and AuNP-KR13 on the HIV-1 virion using TEM. Conclusions: In summary, we report the ability of modified peptide triazole inhibitors that target HIV-1 gp120 to physically disrupt virus particles in the absence of host cells. At conditions similar to those at which both KR13 and AuNP-KR13 conjugates inhibited HIV-1 BaLpseudovirus infection of HOS.T4.R5 cells, it also caused release of HIV-1 gag p24 when incubated with virus alone. Both inhibition of cell infection and p24 release were enhanced substantially by increasing diameter of the multivalent display of KR13 on gold nanoparticles. The mechanism of action of AuNP-KR13 is evidently different from KR13 leading to cell free virus transformation leading to further insights towards virus-cell fusion mechanisms.

Biography:

Ekwu B B Ochigbo is a Registered Pharmacist and a HIV & AIDS resource person. He holds an MPhil with distinction in HIV & AIDS Management from the Africa Centre for HIV & AIDS Management, Stellenbosch University, Cape Town, South Africa. He is also the Director of TibePharmacare, a pharmaceutical and health research consultancy close corporation. His interests are in therapeutics and treatment outcomes research.

Abstract:

The purpose of this research was to determine the level of adherence among HIV infected pregnant women on prevention of mother to child transmission (PMTCT) antiretroviral therapy, and to establish the factors that contribute to poor adherence and their relative importance, in order to suggest intervention strategies that will improve treatment adherence among this population. The study was conducted in Area W Clinic, Francistown Botswana, and was a prospective cross sectional study using semi-structured questionnaire, and data collection form. In total, 61 pregnant women participated in the study and were all within three to nine months gestation, and had been enrolled into the PMTCT program at least more than one month previously. The following were their characteristics: 75% were within the ages of 26 to 42 years old, 90% were single, 81% had attained secondary school education, and 60% were unemployed. Adherence was considered optimum if greater than or equal to 95%. The participants demonstrated a good knowledge of the importance of PMTCT treatment adherence. Reported optimum adherence levels were 84% by virtual analogue assessment, and 82% by pill count. Ninety eight percent of participants reported they did not miss any dose during the last three days before the interview. The most important factors influencing adherence from the study were pregnancy related illnesses, medication side effects, and month of pregnancy of the patient as participants tended to adhere less as they got closer to delivery. It is therefore important for care-givers to carefully monitor patients for these effects, and to carry out continuous adherence counselling with special attention given to those approaching delivery in order to improve or maintain overall adherence to PMTCT therapy. In conclusion, adherence levels to PMTCT therapy among the population sampled was high but can be further enhanced with interventions designed to cover and improve the highlighted areas in the implementation of the preventive therapy.

Biography:

Joan Smith-Sonneborn is a Professor Emeritus who completed her PhD at Indiana University, Postdoctoral studies at Brandeis University, and University of California Berkeley. She was working as a Research Associate, University of Wisconsin, Professor at University Wyoming and did sabbatical training at U Southern California Berkeley, and Monash University, Australia. She has 68 articles in reputed journals, over 100 presentations in United States, England, Germany, Monte Carlo, Sicily, Italy, Japan, Malaysia, Canada, Mexico, and Argentina. She is presently a member of NYAS, GSA, and NSCA. She has served as Chair of the International Ciliate Conference, Gordon Conference, and reviewer for grants, journals, and Advisory Board for FDA.

Abstract:

Telomerase can be touted as the miracle anti-aging enzyme that reverses the age-related attrition of telomere ends. But recent studies reveal a darker side of telomerase related to its role independent of telomere functions in cancer, inherited, and infectious disease states. The catalytic subunit of telomerase, TERT telomerase reverse transcriptase, is known to function as an RNA dependent DNA polymerase in the nucleus (TERT-TERC), as an RNA dependent RNA polymerase, an RdRP (TERT-RMRP), in mitochondria, and, as TERT alone interacting with master regulators of cell function, and/or protection of mitochondrial DNA. The mitochondrial TERT-RMRP is the only mammalian RdRP identified to date, capable of generating double stranded RNA, and acting as a substrate for dicers to generate small interfering RNA’s. TERT also shows conservation of the viral close-right-handed polymerase structure, like viral polymerases, and is capable of producing cDNA, double stranded RNA, and small RNA’s capable of controlling mammalian genome expression. TERT also shows promiscuous partnering with RNA elements, RMPR, tRNA, and TERC. TERT, with its versatile viral-like functions, seems like a valuable hostage for use in viral infection. Telomerase is known to increase in retroviral infections, and can increase mitochondrial oxidative stress resistance by inhibition of pro apoptotic genes, a known role of independent of TERT-TERC. Since telomerase is already a target in anti-cancer drugs, the literature provides a valuable reservoir for potential anti-retroviral drugs. Drugs that inhibit the siRNA’s that promote viral survival, or are necessary for viral survival, offer promising potential success in anti-retroviral therapy, such as interfering with the HIV-TAR miRNA, used to establish initial virus survival and replication or allowing release/stimulation of cell immunity processes using hormetic mimetics.

Biography:

Norbert Nwankwo is working as an assistant professor in Madonna University which is located in Nigeria

Abstract:

Obtaining biological functionalities such as pharmacological activities, disease processes, physiological and structural properties by analyzing the sequence information attributed to them has preliminarily been considered impracticable. This is because there are no known procedures that could be engaged in order to help uncover biological functionalities encoded in the sequence information. It has also been recognized that most biological functionalities of drugs consisting of alkaloids, flavonoids, Terpenes, steroids, etc could be expressed in one gene/protein or the other. For example, Multidrug-resistance transporter gene (MDR1) encoding genes for P-glycoprotein and CYP450, which play vital roles transport and metabolism of Antiretroviral agents have been identified. This MDR1 gene regulates the activity of some antiretroviral drugs. Similarly, anti-bacteria multi-drug resistance genes (MDR1 and MDR2) control the activities of Ciproxfloxacin (an alkaloid), Penicillin (a Beta-lactamase) and tetracycline (a polyketide), Gentamycin (an Aminoglycoside) antibiotics, etc. The same applies to anti-Malaria agents and others. In effect, activities of a wide range of therapeutic agents belonging to numerous groups could be read from one gene/protein they express. Furthermore, genes/proteins encoding therapeutic agents can provide as much information as the therapeutic agents. Direct translations of sequence information into biological functionalities were truly impossible until 1985 when researchers saw proteins/peptides as signals (numerical sequences) instead of piece of fish, meat, bowl of beans or an enzyme. This has opened up a novel area of research. As signals, proteins and peptides were analyzed using techniques that have help develop technologies like Radar, Image Processing and Speech Detector. This technique is called Digital Signal Processing (DSP). This presentation demonstrates how biological functionalities could be translated directly from their sequence information using a DSP technique called Informational Spectrum Method (ISM) and two peptides VIPMFSALS and CAPAGFAIL. This technique has offered direct translation of sequence information into various bio-functionalities. It compared the efficacy of two anti-retroviral agents (Enfurvitude and Sifurvitude) as well as two starter materials (P18 and P32) for the designing of anti-malaria vaccines. It explained the HIV progression to AIDS. It identified the origins of HIV-1 non B subtypes that infected American soldiers on Foreign Service. It elucidated the molecular mechanisms to protection offered to the heart by Influenza vaccines as well as Emilins to Anthrax antigen. It calculated biological functionalities and has helped develop a biomedical device, Computer-Aided Drug Resistance Calculator. Based on the level of biological functionalities uncovered from sequence information using DSP procedures, it can be ascertained that it has now become feasible to obtain same results we get from clinical laboratories by analyzing sequence information involved. Because this approach is rational, it is recommended that it be fully exploited.

Biography:

Namita Kumari is a researcher at Center for Sickle Cell Disease Howard University which is located in USA

Abstract:

Hemin inhibits HIV-1 infection in cultured macrophages and T-cells and also in HIV-1 infected humanized mice by inducing heme oxygenase -1 (HO-1) via a protein kinase C-dependent pathway (reviewed in [1]). HO-1 expression in LPS-treated human macrophages protects them against HIV-1 infection in part through production of MIP1α, MIP1β and LD78β chemokines that decrease CCR5 expression. Iron depletion by iron chelators or through the expression of ferroportin, an iron export protein, inhibited HIV-1 [2, 3]. Here, we show that in heme-treated THP1 cells, mRNA expression of ferroportin, p21, hypoxia-induced factor (HIF)-1α and IKBα, an NF-κB inhibitor was increased and CDK2 expression decreased. HIV-1 replication was also suppressed in THP-1 cells treated with hemin but subsequent treatment with hepcidin restored HIV-1 replication, suggesting that ferroportin played a key role in the HIV-1 inhibition. Stable ferroportin knock-down in THP-1 cells led to the inability of hemin to inhibit HIV-1, further supporting the idea that that ferroportin plays a key role in this process. In addition, hemin treatment reduced the expression of p65 subunit of NF-kB and induced expression of IKBα. Cells treated with hemin were arrested in G1 phase of cell cycle suggesting that expression of p21 and decreased in CDK2 expression affected the cells cycle. Stable HIF-1α knockdown in THP-1 cells increased HIV replication indicating that HIF-1 might also restrict HIV-1 replication. Taken together, our study shows that induction of HIF-1 and iron export pathways might protect hemin-treated THP-1 cells from HIV-1 infection. Additional molecular mechanisms of heme-mediated HIV-1 inhibition might also include NF-kB inhibition by IKBα, reduction of CDK2 epxression and induction of p21 leading to the inhibition of HIV-1 transcription.