Load In HIV Patients Research Articles

Description of Viral Load in HIV Patients Receiving ARVS in Klungkung Regency Hospital

Description of Viral Load in HIV Patients Receiving ARVS in Klungkung Regency Hospital

Stability-indicating method development and validation for the concurrent determination of darunavir, cobicistat, emtricitabine and tenofovir alafenamide by UPLC in bulk and tablet dosage forms

BackgroundTablet dosage forms containing combination of darunavir a protease inhibitor, cobicistat a cytochrome P450 3A inhibitor, emtricitabine and tenofovir alafenamide which were nucleoside reverse transcriptase inhibitors were approved by USFDA on 1st July 2018 to suppress the viral load in HIV patients. It can be used as a complete regimen for the treatment of HIV-1 infection in adults and paediatric patients weighing at least 40 kg. An UPLC method was developed, and separation was done on SB C8 column of dimensions 50 × 2.1 × 1.8 μ with mobile phase 0.01 N potassium dihydrogen ortho phosphate (pH-4.8) and acetonitrile in 60:40 ratio, at a flow rate of 0.3 mL/min and an injection volume of 2 μL. The column temperature was maintained at 30 °C, and detection wavelength was 267 nm. The method was validated according to ICH guidelines.ResultsThe retention times were 1.031, 1.341, 1.630 and 2.153 min, and they were linear in the concentration range of 1.25–7.5 μg/mL, 18.75–112.5 μg/mL, 25–150 μg/mL and 100–600 μg/mL for tenofovir alafenamide, cobicistat, emtricitabine and darunavir, respectively. The intraday and interday precisions were found to be within acceptable limits. LOD was found to be 0.06 μg/mL, 0.51 μg/mL, 1.31 μg/mL and 3.01 μg/mL, and LOQ was 0.19 μg/mL, 1.54 μg/mL, 3.96 μg/mL and 9.13 μg/mL for tenofovir alafenamide, cobicistat, emtricitabine and darunavir. The correlation coefficients were found to be more than 0.999, and recovery was more than 99.52% indicating the method was accurate. Forced degradation studies reveal that the drugs are unstable under acidic conditions. The method was simple, accurate, precise, stable and can be analysed in less runtime of 4 min.ConclusionsThe flexibility, accuracy and precision of the developed method ensure its applicability in routine analysis of tablet dosage forms.Graphical

Cholesterol trajectories and its relationship with viral load in HIV patients over time

Lead Author's Financial Disclosure: Nothing to disclose.

Pruritic papular eruption in HIV-positive patients in Brazil

Pruritic papular eruption (PPE) is a mucocutaneous manifestation associated with HIV infection which was first described in the early years of the HIV epidemic [1]. PPE is characterized by erythematous papules or macules with symmetrical distribution mainly on the trunk and extremities in the absence of other causes of pruritus; intense itching is the most common symptom, leading to scratching, excoriation and postinflammatory hyperpigmentation [2]. PPE tends to be a chronic condition, but individual lesions will appear and disappear. Various aetiological factors have been explored in relation to PPE including immunological responses to insect bites, while infectious agents other than the primary HIV infection do not appear to be implicated [2]. Histopathological and haematological findings have been inconclusive and sometimes inconsistent, with most studies being small [2]. The salient feature of PPE is that it has been strongly and consistently associated with low CD4+ cell count, as described recently in paediatric and adult HIV patients [3,4]. Here, we report the associations that we found between PPE, CD4+ cell count and HIV viral load in HIV patients in Brazil who attended the Hospital Universitário Cassiano Antonio Moraes (HUCAM), Vitória (Espírito Santo) between 2004 and 2010. Patients had a dermatological examination and 4-mm punch skin biopsy, with diagnosis of PPE based on clinical and histopathologic findings to exclude differential diagnosis as scabies, prurigo caused by insect bites, cutaneous drug reactions, lichen planus, acne, demodicosis, eosinophilic folliculitis and bacterial folliculitis [2]. CD4+ cell count and viral load measurements at the time of examination, if available, were obtained from medical records. Ethical permission for use of these data was obtained from HUCAM research ethics committee. Of 370 patients [190 (51.4%) male, 180 (48.7%) female], median age 39 (range 15–82), 17.6% (65/370) had PPE, with no difference in prevalence by age (Pt test = 0.13) or sex (PChi-squared = 0.23). PPE was generalized (more than one body area) in 60.0% (39/65) of cases and PPE manifested mostly [76.9% (50/65)] as papules. Median CD4+ cell count was lower in patients with PPE [266 (interquartile range (IQR) 114–524) cells/μl, n = 45] than without PPE [400 (IQR 238–568) cells/μl, n = 159] (PKruskal–Wallis = 0.02) (Fig. 1a) and viral load was higher [1331 (IQR 5–171797, n = 40) copies/ml compared with 5 (IQR 5–569) copies/ml, n = 133] (PKruskal–Wallis = 0.001) (Fig. 1b); 32.5% (13/40) of patients with PPE had undetectable viral load (<10 copies/ml) compared with 51.9% (69/133) of patients without PPE (PChi-squared = 0.03). Ten patients with PPE were not taking antiretroviral therapy (ART) (15.4%) compared with 56 patients (18.4%) without PPE (PChi-squared = 0.57).Fig. 1: (a) CD4+ cell count in HIV patients with (n = 45) and without (n = 159) pruritic papular eruption [each box indicates the median (horizontal line) and interquartile range, whiskers include data points within 1.5 interquartile range of nearer quartile]; (b) Log10 HIV viral load in HIV patients with (n = 40) and without (n = 133) pruritic papular eruption (each point represents a single measurement, horizontal lines indicate medians).The strong association that we found between low CD4+ cell count and PPE replicates findings in most, but not all [5], previous studies of HIV-related dermatoses. PPE in the early years of the HIV epidemic was considered pathognomonic of AIDS and is potentially prognostic of ART failure [6,7]. Given that the prevalence of PPE has remained relatively unchanged among HIV patients even in the era of ART and that countries such as Brazil continue to see large numbers of new HIV cases each year (44 000 in 2018 http://www.aids.gov.br/pt-br/pub/2019/boletim-epidemiologico-de-hivaids-2019), more consideration needs to be given to identifying effective treatments to relieve debilitating and stigmatizing symptoms of HIV-related dermatological conditions and to improve patient quality of life [2]. Acknowledgements We thank Professor Elton Lucas and Professor Tania Reuter from the Hospital Universitário Cassiano Antonio Moraes (HUCAM) for their advice. This study received no specific funding. Conflicts of interest The authors have no conflicts of interest to declare.

Application of Structure-based Methods to Analyze Resistance Mutations for Chemically Diverse Non-Nucleoside Reverse Transcriptase Inhibitors.

Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are used in combination with antiretroviral therapy to suppress viral loads in HIV patients. The chemical design of NNRTIs has changed in recent years in response to resistance-associated mutations (RAMs) and resistance. NNRTIs are chemically diverse compounds that bind an allosteric site of HIV RT. Resistance- associated mutations (RAMs) identified in HIV patients are associated with NNRTI resistance. RAMs confer amino acid changes that alter both structural and physiochemical properties of the allosteric site. Ultimately, these changes reduce NNRTI affinity. Previously, we used a combination of computational and experimental methods to analyze and validate RAMs for 3 diarylpyrimidine (DAPY) NNRTIs. The objective of this study is to apply these methods to other chemically diverse, non- DAPY NNRTIs. We selected MIV-150 (experimental microbicide) and doravirine for this study. A computational and molecular modeling strategy was used to evaluate the effects of RAMs. Calculated changes in drug affinity and stability (ΔS + ΔA) were used to determine overall resistance levels: susceptible, low, intermediate, and high. The ΔS + ΔA values for K101P suggest that this mutation confers intermediate/high-level resistance to MIV-150, but remains susceptible to doravirine. Based on the determined resistance levels, we analyzed the models and used Molecular Dynamics (MD) to compare the interactions of MIV-150/doravirine with RT wild-type (WT) and RT (K101P). From MD, we found that key interactions were lost with RT (K101P), but were retained with doravirine. To experimentally validate our findings, we conducted a fluorescence-based reverse transcription assay for MIV-150 with RT (WT) and RT (K101P). IC50 values determined in assays showed a 101-fold change in potency for MIV-150, but essentially no change for doravirine. Our computational and experimental results are also consistent with antiviral data reported in the literature. We believe that this approach is effective for analyzing mutations to determine resistance profiles for chemically diverse NNRTIs in development.

The Adherence to Fixed-Dose Combination Antiretroviral Therapy and Viral Load in HIV Patients

Antiretroviral fixed-dose combination (FDC ARV) has been used in HIV patient therapy. The therapy is recommended to be monitored using a viral load (VL), reflecting the amount of HIV in the blood. Since 2015, the VL testing has been carried out at Pengayoman Cipinang Hospital. However, the evaluation of adherence to the FDC antiretroviral therapy on VL has never been conducted. This study aims to determine the relationship between the adherence level to the FDC ARVs and VL at Pengayoman Cipinang Hospital in 2015-2018. The study used a cross-sectional design and a random sampling technique. The sample size was 70, and data generated from the medical record. Kruskal Wallis test showed no differences in the level of adherence in the group based on gender, education level, marital status, employment status, and the risk factor for HIV. However, there was a significant difference in the age group. Cross-tab data showed virological failure (VL> 400 copies/ml) was found in 1.8% of patients with good adherence, 75% of patients with moderate adherence, and 100% of patients with poor adherence. The Chi-square test showed a significant relationship between adherence to FDC ARV and VL (p=0.000). It can be concluded that there was a significant relationship between the level of adherence in taking ARV FDC and viral load.

Association between Parvovirus B19 and anemia in HIV-infected patients.

Background: Human parvovirus B19 (B19V) can cause anemia in some patients, including those with compromised immunity system. There are a few studies on molecular epidemiology of B19V and its association with anemia in Iran. Therefore, the aim of this study was to determine the B19V DNA, IgM, IgG, genotyping, and viral load in HIV patients in different groups of pregnant women, general population, injection drug users (IDU), and Elite controllers. Also, the possible association of B19V with anemia was studied. Methods: In this case-control study, B19V DNA, anti-B19V IgM, anti-B19V IgG, viral load, and hemoglobin level were assessed in 113 HIV positive patients and 72 healthy controls. Also, CD4+ T cell counts and HIV load were measured in the patients’ group. All statistical analyses were done using STATA 14.2 software (Stata Corporation, College Station, Texas, USA). P value < 0.05 was considered statistically significant. Results: Among HIV patients, 19 (16.8%) cases had B19V DNA, 3 (2.7%) had B19V IgM, and 7 (6.2%) had B19V IgG. In control group, the prevalence of B19V DNA, IgM, and IgG was 6 (8.33%), 7(9.7%), and 19 (26.4%), respectively. In subpopulations based on transmission routes, general population had the highest B19V IgG and DNA positivity prevalence and viral load level. There was no significant association between B19V antibodies and DNA with anemia. Conclusion: The results demonstrated that B19V infection cannot be considered as a high-risk factor for anemia in adult HIV patients. However, further studies are needed to determine the exact role of B19V infection in HIV patients.

KORELASI ANTARA KADAR INTERFERON-γ PLASMA DENGAN JUMLAH VIRAL LOAD DI PENDERITA HIV

The incidence of HIV is increasing in Indonesia and Asia as well, Indonesia is considered as the most rapid. One of the diagnostic toolsfor diagnosing HIV is by viral load. Lymphocyte T-CD8+ secreted IFN-γ will inhibit replication of HIV virus through the induction of antiviralprotein and the host immune response, which kills infected cells. An examination of plasma IFN-γ and viral load will be more convincingfor the treatment and/or to know the progressiveness of HIV &amp; AIDS. The aim of this study is to know the correlation between plasma IFN-γand viral load in HIV patients. Forty two samples from HIV patients were collected at the Intermediate Care and Infectious Disease Unit ofDr. Soetomo Hospital, Surabaya from April to June 2011. The concentration of plasma IFN-γ was measured by ELISA (eBioscience) methodand the amount of viral load was measured using PCR Cobas Amplicor (Roche Diagnostics). The level of plasma IFN-γ in this study wasfound 11.4 pg/mL up to 576 pg/mL and the level of viral load was 589 copies/mL up to 510.000 copies/mL. The statistical analysis showedno significant correlation (p&gt;0.05) between plasma IFN-γ level and viral load in HIV patients, and no correlation was found between IFN-γplasma and viral load in HIV patients.

Bacterial etiology and antibiotic resistance pattern of septicemia in HIV and non-HIV patients admitted to tertiary care hospitals, Shiraz, South of Iran

The present study aimed to determine the bacteriological etiology and antibiotic susceptibility pattern of sepsis in HIV infected and HIV uninfected patients, and related risk factors to introduce an appropriate therapy. This cross-sectional study was conducted from January 2014 to January of 2015 enrolling patients with sepsis associated with or without HIV infection admitted to Shiraz teaching hospitals, South of Iran. Blood and urine cultures were performed and standard microbiological methods were followed for isolation and identification of the bacteria. HIV antibody testing and CD4+ lymphocyte count were done for HIV-infected patients. Antimicrobial susceptibility tests were performed using the disk diffusion method in accordance with CLSI recommendations. Totally, 140 patients with sepsis including 30 HIV-positive, and 110 HIV-negative were enrolled. Our finding showed 26.7% and 20% blood and urine culture positivity in HIV-positive and 20.9% and 14.5% positivity in HIV-negative patients. Staphylococcus aureus, Salmonella spp. and coagulase-negative staphylococci (CoNS) each with frequency of 25% were detected as the most prevalent isolates in samples of HIV patients. In contrast, the main etiology for sepsis in HIV-negative patients was CoNS (47.8%), followed by Escherichia coli (17.4%). The median of CD4+ lymphocyte count and viral load in HIV patients were estimated 10.15 cells/mm3 and 68019.48 copies/mL, respectively. The results of the present study revealed that the main cause of sepsis in the studied hospitals was nosocomial pathogens. These findings highlighted the importance of infection control policies for preventing the emergence and spread of nosocomial infections.

HIV-1 Tat exacerbates lipopolysaccharide-induced cytokine release via TLR4 signaling in the enteric nervous system.

The loss of gut epithelium integrity leads to translocation of microbes and microbial products resulting in immune activation and drives systemic inflammation in acquired immunodeficiency syndrome (AIDS) patients. Although viral loads in HIV patients are significantly reduced in the post-cART era, inflammation and immune activation persist and can lead to morbidity. Here, we determined the interactive effects of the viral protein HIV-1 Tat and lipopolysaccharide (LPS) on enteric neurons and glia. Bacterial translocation was significantly enhanced in Tat-expressing (Tat+) mice. Exposure to HIV-1 Tat in combination with LPS enhanced the expression and release of the pro-inflammatory cytokines IL-6, IL-1β and TNF-α in the ilea of Tat+ mice and by enteric glia. This coincided with enhanced NF-κB activation in enteric glia that was abrogated in glia from TLR4 knockout mice and by knockdown (siRNA) of MyD88 siRNA in wild type glia. The synergistic effects of Tat and LPS resulted in a reduced rate of colonic propulsion in Tat+ mice treated with LPS. These results show that HIV-1 Tat interacts with the TLR4 receptor to enhance the pro-inflammatory effects of LPS leading to gastrointestinal dysmotility and enhanced immune activation.

Monitoring the Viral Load in HIV Patients under Anti-Retroviral Therapy ( Art )

Viral load monitoring is an important factor in managing HIV disease. Antiretroviral therapy is the recommended treatment for HIV patients, and the goal therapy is achieving viral suppression and reducing viral load below the level of detection. Viral load is an important parameter used to monitor the progression of HIV and critically regarding treatment decision. The results of present study revealed that there were statistical significant differences between patients maintained on treatment (GA) and patients without treatment (GB) regarding the viral load, and clearly indicated that adherence to ART playing a role in suppression the viral load supporting the immune system of HIV infected patients. The success of ART for someone living with HIV depends on, starting the treatment at the right time, choosing the right combination of (ART) and monitoring the effectiveness of the treatment through monitoring the vilral load.

High heritability is compatible with the broad distribution of set point viral load in HIV carriers.

Set point viral load in HIV patients ranges over several orders of magnitude and is a key determinant of disease progression in HIV. A number of recent studies have reported high heritability of set point viral load implying that viral genetic factors contribute substantially to the overall variation in viral load. The high heritability is surprising given the diversity of host factors associated with controlling viral infection. Here we develop an analytical model that describes the temporal changes of the distribution of set point viral load as a function of heritability. This model shows that high heritability is the most parsimonious explanation for the observed variance of set point viral load. Our results thus not only reinforce the credibility of previous estimates of heritability but also shed new light onto mechanisms of viral pathogenesis.

No reduction of HCV viral load in HIV patients co-infected with HCV genotype 1 during a 30 days course of nitazoxanide monotherapy

There are two new drugs approved and several in development for treatment of chronic HCV; among them nitazoxanide (NTZ). Twelve HIV/HCV genotype 1 co-infected patients were enrolled prospectively to receive a 30days course of oral NTZ 500mg bid. This therapy was well tolerated in this group of HIV patients co-infected with HCV genotype 1. Nevertheless no changes in HCV viral load were observed during treatment in none of the patients evaluated. This data suggests that despite the promising results reported for HCV genotype 4 mono-infected patients, NTZ exhibit poor activity as monotherapy in HIV/HCV co-infected patients with genotype 1.

Pneumocystis-associated organizing pneumonia as a manifestation of immune reconstitution inflammatory syndrome in an HIV-infected individual with a normal CD4+ T-cell count following antiretroviral therapy

We describe a rare case of Pneumocystic jirovecii-associated organizing pneumonia (PJP) in an HIV-infected individual on highly active antiretroviral therapy (HAART) with a CD4(+) T-cell count of 835 x 10(3) cells/mL and a low viral load. PJP was confirmed using transbronchial biopsies and bronchoalveolar lavage. The presentation in this patient suggests immune reconstitution inflammatory syndrome (IRIS) after institution of antiretroviral therapy (ART). This case report, however, is the first documented presentation of PJP in a patient with CD4 count greater than 300 prior to the induction of HAART who developed PJP and organizing pneumonia as a manifestation of IRIS. This suggests that there is continuing immune dysfunction in the face of re-expansion of CD4(+) T-cells and low viral load in HIV patients despite ART.

A Tolerance Interval Approach for Assessment of Agreement with Left Censored Data

We describe a tolerance interval approach for assessing agreement in method comparison data that may be left censored. We model the data using a mixed model and discuss a Bayesian and a frequentist methodology for inference. A simulation study suggests that the Bayesian approach with noninformative priors provides a good alternative to the frequentist one for moderate sample sizes as the latter tends to be liberal. Both may be used for sample sizes 100 or more, with the Bayesian one being slightly conservative. The proposed methods are illustrated with real data involving comparison of two assays for quantifying viral load in HIV patients.

Stata Tip 1: The Eform() Option of Regress

Did you know about the eform() option of regress? It is very useful for calculating confidence intervals for geometric means and their ratios. These are frequently used with skewed Y -variables, such as house prices and serum viral loads in HIV patients, as approximations for medians and their ratios. In Stata, I usually do this by using the regress command on the logs of the Y -values, with the eform() and noconstant options. For instance, in the auto dataset, we might compare prices between non-US and US cars as follows:

HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing CD36-dependent cholesteryl ester accumulation in macrophages

Protease inhibitors decrease the viral load in HIV patients, however the patients develop hypertriglyceridemia, hypercholesterolemia, and atherosclerosis. It has been assumed that protease inhibitor-dependent increases in atherosclerosis are secondary to the dyslipidemia. Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of CD36 and the accumulation of cholesteryl esters. The use of CD36-blocking antibodies, a CD36 morpholino, and monocytes isolated from CD36 null mice demonstrated that protease inhibitor-induced increases in cholesteryl esters were dependent on CD36 upregulation. These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating CD36 and cholesteryl ester accumulation independent of dyslipidemia. Studies with LDL receptor null mice demonstrated that low doses of protease inhibitors induce an increase in the level of CD36 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids. Furthermore, the lack of CD36 protected the animals from protease inhibitor-induced atherosclerosis. Finally, ritonavir increased PPAR-gamma and CD36 mRNA levels in a PKC- and PPAR-gamma-dependent manner. We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of CD36 and the subsequent accumulation of sterol in macrophages.

HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing CD36-dependent cholesteryl ester accumulation in macrophages

Protease inhibitors decrease the viral load in HIV patients, however the patients develop hypertriglyceridemia, hypercholesterolemia, and atherosclerosis. It has been assumed that protease inhibitor–dependent increases in atherosclerosis are secondary to the dyslipidemia. Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of CD36 and the accumulation of cholesteryl esters. The use of CD36-blocking antibodies, a CD36 morpholino, and monocytes isolated from CD36 null mice demonstrated that protease inhibitor–induced increases in cholesteryl esters were dependent on CD36 upregulation. These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating CD36 and cholesteryl ester accumulation independent of dyslipidemia. Studies with LDL receptor null mice demonstrated that low doses of protease inhibitors induce an increase in the level of CD36 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids. Furthermore, the lack of CD36 protected the animals from protease inhibitor–induced atherosclerosis. Finally, ritonavir increased PPAR-γ and CD36 mRNA levels in a PKC- and PPAR-γ–dependent manner. We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of CD36 and the subsequent accumulation of sterol in macrophages.

Strong correlation between the complement-mediated antibody-dependent enhancement of HIV-1 infection and plasma viral load.

We have previously demonstrated that complement-mediated antibody-dependent enhancement (C-ADE) of HIV-1 infection correlates with accelerated immunosuppression and disease progression in HIV-1-infected individuals. In the present work the relationship between C-ADE and plasma HIV-1 RNA concentrations was studied to determine the effect of C-ADE on viral replication. Three studies were performed: (a) C-ADE and HIV-1 RNA concentrations were determined in the serum and plasma aliquots taken at the same time from 98 HIV patients, mostly in the advanced stage of the disease; (b) the above two parameters as well as HIV enzyme-linked immunosorbent assay (ELISA)-reactive antibodies (Abbott HIV 1/2 test), and p24 antigen levels (Abbott antigen test; Abbott, Delkenheim, Germany) were determined in four seroconversion panels purchased from the Boston Biomedica firm; (c) changes of HIV-1 RNA concentration and C-ADE during a 17 month follow-up period were determined in 18 HIV-infected patients. C-ADE was measured by the method previously established in our laboratories. The results were expressed by an enhancement/neutralization index (E/NI). HIV-1 RNA levels were determined with the Amplicor monitor kit (Roche, Basel, Switzerland), and in some experiments with the nucleic acid sequence based amplification (Organon Teknika, Turnhout, Belgium) kits. (a) We found a highly significant (P<0.0001) positive correlation between E/NI values reflecting the extent of HIV-1 infection enhancement and plasma HIV-1 RNA levels. Both E/NI and HIV-1 RNA levels negatively correlated to the CD4 cell counts. (b) C-ADE was first detected just before, or concomitantly with, seroconversion in 4/4 seroconversion panels. (c) Both E/NI values and HIV-1 RNA levels significantly (P<0.001) increased during a 17 month observation period in 18 HIV-infected patients. We found strong association between the extent of the complement-mediated antibody-dependent enhancement of HIV-1 infection and the plasma viral load in HIV patients. On the basis of these findings, C-ADE correlates with HIV replication in vivo, and potentially contributes to the progression of HIV disease.

Variation in body weight and plasma viral load in HIV patients treated with tritherapy including a protease inhibitor

Variation in body weight and plasma viral load in HIV patients treated with tritherapy including a protease inhibitor