bluuebunny/crossref_metadata_2025_split huggingface dataset:
Everything fine so far? Physical and mental health in HIV‐infected patients with virological success and long‐term exposure to antiretroviral therapy
Authors: Erdbeer Gesa , Sabranski Michael , Sonntag Ina , Stoehr Albrecht , Horst Heinz‐A. , Plettenberg Andreas , Schewe Knud , Unger Stefan , Stellbrink Hans‐J. , Fenske Stefan , Hoffmann Christian 
Published: November 2014

IntroductionLittle is known about the well‐being on long‐term exposure to antiretroviral therapy. The ACTG Augmented Symptoms Distress Module (ASDM) is a validated tool which measures the presence of a total of 22 symptoms seen with HIV and quantifies the extent to which they cause distress to the patient.MethodsELBE was a cross‐sectional study that consecutively included adult HIV‐infected patients presenting with viral suppression (<50 HIV RNA copies/mL) and ART exposure for at least five years. Patients were evaluated by four different questionnaires, including ASDM.ResultsOf a total of 894 patients included in the three participating ELBE centres, complete data on ASDM were available for 698 patients (626 male, 69 female, 3 transsexual). Median age was 49.7 years (range, 23.3–82.5 years) and median exposure to ART was 11.5 years (range, 5–28 years). Median CD4 T‐cell counts had increased from a CD4 nadir of 180 to currently 640 cells/µL. Despite immunological and virological success, a high degree of symptom‐related distress was noted in this patient population. In total, 63.8% and 36.3% of the patients had at least one “bothersome” or one “very bothersome” symptom, respectively. The symptoms most frequently reported to be “bothersome” or “very bothersome” were fatigue and energy loss (18.5% and 11.0% respectively), insomnia (12.8% and 11.6%), sadness and depression (13.0% and 10.0%), sexual dysfunction (12.0% and 10.0%), and changes in body appearance (11.0% and 10.9%). There was no association between the degree of symptom‐related distress and gender, age or CD4 T‐cell nadir. However, the history of AIDS‐defining illnesses, comorbidities such as depression but also the duration of ART were significantly associated with a higher overall symptom summary score and with a higher frequency of symptoms. For example, in patients with at least 15 years of ART exposure, only 27.3% of the patients did not report at least one “bothersome” or “very bothersome” symptom.ConclusionsIn this large group of positively selected HIV+ patients with virological success and long‐term exposure to ART, a high degree of symptom‐related distress was found. Medical care of HIV‐infected patients should not only focus on optimal virological outcome. More data on quality of life in patients with long‐term exposure to ART is needed.
Detection of resistance mutations and CD4 slopes in individuals experiencing sustained virological failure
Authors: Schultze Anna , Paredes Roger , Sabin Caroline , Phillips Andrew N , Pillay Deenan , Kirk Ole , Lundgren Jens D , Pozniak Anton , Nelson Mark , Cozzi‐Lepri Alessandro ,   on behalf of Eurosida in Eurocoord
Published: November 2014

IntroductionSeveral resistance mutations have been shown to affect viral fitness, and the presence of certain mutations might result in clinical benefit for patients kept on a virologically failing regimen due to an exhaustion of drug options. We sought to quantify the effect of resistance mutations on CD4 slopes in patients undergoing episodes of viral failure.Materials and MethodsPatients from the EuroSIDA and UK CHIC cohorts undergoing at least one episode of virological failure (>3 consecutive RNA measurements >500 on ART) with at least three CD4 measurements and a resistance test during the episode were included. Mutations were identified using the IAS‐US (2013) list, and were presumed to be present from detection until the end of an episode. Multivariable linear mixed models with a random intercept and slope adjusted for age, baseline CD4 count, hepatitis C, drug type, RNA (log‐scale), risk group and subtype were used to estimate CD4 slopes. Individual mutations with a population prevalence of >10% were tested for their effect on the CD4 slope.ResultsA total of 2731 patients experiencing a median of 1 (range 1–4) episodes were included in this analysis. The prevalence of any resistance per episode was 88.4%; NNRTI resistance was most common (78.5%). Overall, CD4 counts declined by 17.1 (−19.7; −14.5) cells per year; this decline was less marked with partial viral suppression (current HIV RNA more than 1.5 log below the setpoint; p=0.01). In multivariable models adjusting for viral load, CD4 decline was slower during episodes with detected resistance compared to episodes without detected resistance (21.0 cells/year less, 95% CI 11.75–30.31, p<0.001). Among those with more than one resistance mutation, there was only weak evidence that class‐specific mutations had any effect on the CD4 slope (Table 1). The effects of individual mutations (incl. M184V) were explored, but none were significantly associated with the CD4 slope; for these comparisons, a Bonferroni‐corrected p‐value level was 0.003.ConclusionsIn our study population, detected resistance was associated with slightly less steep CD4 declines. This may be due to a biological effect of resistance on CD4 slopes, or other unmeasured factors such as poor adherence among individuals without resistance. Among individuals with detected drug resistance, we found no evidence suggesting that the presence of individual mutations was associated with beneficial CD4 slope changes.
Appearance of NS3 Q80K mutation in HCV genotype 1a mono‐ or HIV/HCV co‐infected patients in a Berlin laboratory
Authors: Ehret Robert , Neifer Stefan , Walter Hauke , Baumgarten Axel , Obermeier Martin 
Published: November 2014

IntroductionSimeprevir, a new oral NS3/4A protease inhibitor, was recently approved by the FDA and the EMA for the treatment of patients with chronic HCV genotype 1, 4, 5 and 6 infection l. It has been recommended in the 2014 UK Consensus Guidelines as a possible treatment of previously untreated genotype 1a‐infected patients. The antiviral efficacy of simeprevir is adversely affected by the mutation at the Q80K loci. There is controversial discussion that the incidence of Q80K in the European HCV 1a‐infected community is very low and therefore testing of Q80K before starting a therapy including simeprevir is not necessary. We analyzed the appearance of Q80K in all sequenced HCV NS3A samples in 2014 in our laboratory.Materials and MethodsAll in 2014 received orders for HCV resistance tests were analyzed with an in‐house bulk sequencing method analyzing NS3A amino acids 1–181. Analysis was performed using geno2pheno HCV. The genotype 1a samples were selected, Q80K status and data of HIV co‐infection were collected.ResultsForty‐two HCV 1a samples were sent to us for resistance analyses from nine different medical centres in Berlin and Hannover, Germany. Nineteen (or 45%) of the sequences showed a Q80K mutation. Six extra clade I viruses had no Q80K mutation. Comparison between mono‐ and HIV‐1 co‐infected patients showed no difference in frequency of Q80K (mono‐infected: 8 out of 19 patients; co‐infected: 9 out of 23). For two 80K‐positive patients, the HIV‐status was not available.ConclusionsThe incidence for Q80K mutation in HCV genotype 1a with overall 45% is substantially high in our cohort and does not differ between mono‐ and HIV‐1 co‐infected patients. Response to simeprevir is affected by the presence of viral Q80K. When treating HCV‐infected patients with a simeprevir containing regimen, it is therefore important that HCV does not contain the Q80K mutation.
New dolutegravir resistance pattern identified in a patient failing antiretroviral therapy
Authors: Carganico Andreas , Dupke Stefan , Ehret Robert , Berg Thomas , Baumgarten Axel , Obermeier Martin , Walter Hauke 
Published: November 2014

IntroductionThe most recently approved antiretroviral, the integrase inhibitor dolutegravir (DTG), is described to be a very potent drug with a unique resistance profile, but a certain degree of cross‐resistance to RAL or EVG induced drug resistance, which is mediated mainly by integrase mutations at positions 140 and 148. The impact of a single N155H mutation to DTG resistance is described to be minor. However, there is only rare data available about the impact of N155H in the context of secondary site integrase mutations. Here, we present a case of virological failure in a DTG treated patient based on N155H mutation background.MethodsTherapy monitoring of an HIV‐HCV co‐infected patient harbouring already an omni‐drug‐class resistant HIV‐1 in consequence of more than 20 years ART history. Drug susceptibility testing was performed by RT‐PCR from plasma and subsequent Sanger sequencing. Tropism testing was done from proviral DNA with FPR cut‐offs according to the German recommendations.ResultsIn 2013, the patient harbouring a virus with high level resistance to all RTI and PI received a regimen containing FTC, TDF, DRV/r, RPV, T20, and RAL to handle a viral load of 5000 RNA copies/mL, but never achieved fully suppressed viral load. In June 2013, after S119R, N155H and E157Q mutations in viral integrase were detected, the patient received DTG, and RAL was stopped. One month later, when viral load was undetectable for the first time since 2007, ART was de‐escalated by removing T20. Since February 2014, low‐level viral load was re‐detectable. Two new mutations T97A and S147G in the integrase and no other new resistance associated mutations in protease and reverse transcriptase in comparison to the sample analyzed in June 2013 were detected.ConclusionsHighly resistant HIV‐1 strains have been a common problem in the past. Their frequencies were pushed back by highly potent ART, but the virus is still able to become resistant against all available antiretrovirals at once. The here documented strain became resistant to DTG without carrying mutations at the described positions 140 and 148, but in the context of integrase N155H. Since N155H alone is described not to contribute sufficient resistance to DTG, there seems to be a need to re‐check viruses with N155H plus minor mutations (like T97A, S119R, S147G and E157Q) potentially contributing to DTG resistance.
Determinants of use of the fixed dose combination emtricitabine/rilpivirine/tenofovir (Eviplera) in HIV‐infected persons receiving care in Italy
Authors: Cozzi‐Lepri Alessandro , Lo Caputo Sergio , Maggiolo Franco , Antinori Andrea , Ammassari Adriana , Marchetti Giulia , Mastroianni Claudio , Gori Andrea , Di Perri Giovanni , Angarano Gioacchino , Carbone Alessia , d'Arminio Monforte Antonella ,   Study, on behalf of the ICONA Foundation
Published: November 2014

IntroductionEmtricitabine/rilpivirine/tenofovir (EVP) is a fixed‐dose combination of antiretrovirals (ARV) approved by the European Medicines Agency in November 2011 and introduced in Italy in February 2013. It is a once‐a‐day single tablet and is licensed in Europe for use only in ARV‐naïve patients with a viral load (VL) ≤100,000 copies/mL.ObjectiveTo identify factors that may be associated with the use of EVP as first‐line regimen in HIV‐infected individuals starting cART from ARV‐naïve in Italy.MethodsClinical sites in ICONA Foundation Study in which ≥1 person had started EVP were selected for this analysis. From these we included all patients who started an EVP‐based cART regimen as well as those starting other cART regimens after the date of introduction of EVP at the site (after February 2013 in any case) and with a VL ≤100,000 copies/mL from ARV‐naïve. Characteristics at the time of starting cART were compared using chi‐square test and unadjusted and adjusted logistic regression analysis. Factors investigated included: gender, mode of HIV transmission, time from HIV diagnosis, CD4 count, nation of birth, AIDS, HCV‐status, age, CD8 count, VL, diabetes, smoking, total and HDL cholesterol, eGFR, blood glucose, level of education and employment and site location. Factors showing unadjusted associations with a p‐value of 10% or smaller, were retained in the multivariable model.ResultsWe identified 183 patients starting EVP and 173 starting the control regimen from 23 sites. The number of patients starting EVP included at each site ranged from 1 to 12 and the number of those starting the control regimen was similar. The most frequently used drugs in the concurrent group were: TDF (75%), FTC (74%), DRV (39%), ATV/r (26%), LPV/r (9%), EFV (13%) and RAL (14%). In univariable analysis, there were differences in median CD4 count (390 cells/mm3 in EVP versus 348 in controls, p=0.002), time from HIV diagnosis to starting cART (11 versus 3 months, p=0.001) and prevalence of students (6% versus 3%, p=0.07). No differences were observed for all other factors examined. The table shows estimates of the odds ratios (OR) for factors included in the multivariable model.ConclusionsCD4 count was higher in EVP‐treated patients compared to controls. Guidelines suggest avoiding initiation of EVP in presence of high VL, possibly explaining this residual difference in CD4. There was also a tendency to prescribe EVP to people with perceived lower adherence or hesitant to start or perhaps with a slow progressing disease.
Efficacy of PI monotherapy versus triple therapy for 1964 patients in 10 randomised trials
Authors: Arribas Jose , Girard Pierre‐Marie , Paton Nicholas , Winston Alan , Marcelin Anne‐Genevieve , Elbirt Daniel , Hill Andrew , Blanca Hadacek Maria 
Published: November 2014

IntroductionThe efficacy of protease inhibitor monotherapy has been analyzed with different endpoints: initial HIV‐1 RNA rebound, long‐term HIV‐1 RNA suppression after re‐intensification, treatment‐emergent drug resistance, neurocognitive testing and HIV‐1 RNA in the cerebrospinal fluid (CSF).MethodsA PUBMED search identified nine randomised trials of PI monotherapy versus triple therapy in patients with HIV RNA<50 copies/mL at baseline. Results from the recently completed PROTEA trial were also included. The percentage of patients with HIV RNA suppression <50 copies/mL was analyzed using switch equals failure and intensification included endpoints (ITT). The number of patients with new drug resistance mutations, HIV RNA in the CSF or change in neurocognitive function was analyzed by treatment arm.ResultsFour trials evaluated darunavir/r monotherapy (n=785: MONET, MONOI, MONARCH, PROTEA), five evaluated lopinavir/r monotherapy (n=592: OK‐04, KalMo, KALESOLO, KRETA, MOST) and one evaluated both (MRC PIVOT, n=587). HIV‐1 RNA suppression rates tended to be lower on PI monotherapy than triple therapy in “switch equals failure” analysis (76% vs 82%), but not when the outcome of intensification was included (87% vs 85%). There were small numbers of patients taking PI monotherapy with detectable HIV‐1 RNA in the CSF, in three trials: PROTEA (n=2), MONOI (n=2) and MOST (n=5), but only two cases of CSF viral escape (MONOI). In two trials, there was no difference in neurocognitive test results between PI monotherapy and triple therapy, based on z‐scores from five domains (in PROTEA, mean change in NPZ‐5 score=0.0 for DRV/r monotherapy vs −0.10 for triple therapy); similar results were observed in the MRC PIVOT trial. The risk of treatment emergent NRTI or PI resistance (Table 1) was 11/973 (1.1%) for patients on PI monotherapy, versus 7/991 (0.7%) for patients on triple therapy.ConclusionsIn 10 randomised trials of 1964 patients with HIV‐1 RNA suppression at baseline, PI monotherapy showed a higher risk of HIV RNA elevations, and small numbers with HIV RNA detectable in CSF and concomitantly in the plasma. However there was no increased risk of treatment‐emergent drug resistance, neurocognitive endpoints were not different between the arms and HIV‐1 RNA suppression rates after intensification were similar between PI monotherapy and triple therapy.
Durability of lopinavir/ritonavir dual‐therapies in individuals with viral load <50 copies/mL in the observational setting
Authors: Gianotti Nicola , Cozzi‐Lepri Alessandro , Antinori Andrea , Di Biagio Antonio , Cristina Moioli Maria , Nozza Silvia , Cingolani Antonella , De Luca Andrea , Madeddu Giordano , Bonora Stefano , Ceccherini‐Silberstein Francesca , d'Arminio Monforte Antonella ,   Study on behalf of ICONA Foundation
Published: November 2014

IntroductionWe aimed at evaluating the efficacy and durability of a lopinavir/ritonavir‐based dual regimen (LPV/r‐DR) in virologically controlled HIV‐infected individuals in current clinical practice.MethodsPatients who have initiated for the first time a LPV/r‐DR with HIV‐RNA<50 copies/mL were included in this observational study. The main endpoints were: time to virological rebound [VR=time of first of two consecutive viral loads (VL)>50 copies/mL] and time to experience either a single VL>200 copies/mL or discontinuation/intensification (= treatment failure, TF). Individuals’ follow‐up accrued from the date of starting the LPV/r‐DR to event or last available VL. Kaplan‐Meier curves and Cox regression analysis were used. Covariates included in the multivariable analysis were gender, age, route of transmission, hepatitis co‐infection, calendar year of starting the DR, nadir CD4+ count, VL at initiation of first cART, previous failures to protease inhibitors (PIs), time with undetectable VL before starting the DR and the type of DR [nucleoside reverse transcriptase (NRTI), non‐NRTI (NNRTI), raltegravir or maraviroc, with NRTI as reference group]. Results are presented as median (Q1, Q3) or frequency (%) as appropriate.Results108 individuals followed for 18 (7, 30) months were included; baseline (BL) characteristics are detailed in Table 1.LPV/r was associated with a NRTI in 51, with a NNRTI in 10, with raltegravir in 29, and with maraviroc in 18 individuals. By 36 months from switching to the LPV/r‐DR, the proportion of individuals with VR and TF was 10% (95% CI 3–17%) and 36% (95% CI 22–50%), respectively. We did not find any factor independently associated with the risk of VR. Older age (ARH=0.49 (95% CI 0.30–0.78) per 10 years older; p=0.003) was found to be protective from TF. Mean (SE) CD4+ cells/µL increase from BL to month 36 resulted significant: 195 (40.1) cells/µL (p=0.0028). We did not observe significant changes in AST, ALT, eGFR (MDRD formula), triglycerides and both total and HDL‐cholesterol.ConclusionsA LPV/r‐DR can be considered a valuable option in patients with HIV‐RNA<50 copies/mL and ongoing toxicity from the third drug of the regimen, although up to 17% of patients showed viral rebound by 3 years. Older patients are at lower risk of failure with this strategy, but larger sample size is needed to identify who might benefit from this strategy instead of others.
Lamivudine plus darunavir boosted with ritonavir as simplification dual regimen in HIV‐infected patients
Authors: Luis Casado Jose , Bañón Sara , Moreno Ana , Diaz de Santiago Alberto , Gomez Cristina , Perez‐Elías María J , Moreno Santiago 
Published: November 2014

IntroductionThe combination of lamivudine plus a protease inhibitor boosted with ritonavir (PI/r) has become an alternative as simplification strategy in HIV‐infected patients with toxicity/intolerance to other nucleoside analogues (NA). Lamivudine plus darunavir/r (DRV/r) could be an adequate once daily option.Materials and MethodsProspective cohort study of 48 HIV‐infected patients on suppressive triple therapy‐based HAART, HBV negative, who switched to lamivudine 300 mg plus DRV/r 800/100 mg once daily.ResultsMean age was 50 yrs (35–74), and 65% were male. Thirty patients (63%) had HCV co‐infection (fibrosis 4 in 7 cases, 23%). Median time of HIV infection was 19.1 years, and CD4+ count nadir was 220 cells/µL (2–604). They had received a mean of three regimens before (2–20), and 20 (42%) had a previous AIDS diagnosis. In eight cases, a previous resistance test showed two to seven secondary mutations in the protease gene, without resistance to DRV/r (one patient with the I84V mutation). At baseline, patients had viral suppression (<50 copies/mL) for a median time of 1263 days (341–1884), and they were receiving predominantly a PI based regimen (ATV in four, FPV in four, LPV in three, DRV in six) or an efavirenz‐based regimen (seven). The main reason to switching to this dual therapy was toxicity (35 patients, 75%), mainly renal toxicity attributed to tenofovir (24 cases). During 104.3 patients‐year of follow‐up (median 912 days), only two patients (4%) failed at 27 and 505 days, due to non‐adherence and lost to follow up, respectively. Total cholesterol and triglycerides increased significantly during the first six months after initiation (TC, from 185 to 269 mg/dL; p=0.01, TG from 118 to 185 mg/dL; p=0.03, TC/HDL ratio, from 4.09 to 4.66) and decreased after. Median estimated glomerular filtration rate (eGFR) improved during follow up (from 86 to 96.1 mL/min; p=0.13). In patients with renal toxicity as cause of switch there was a mild, no significant improvement during the first year (from 63.3 to 68.7 mL/min), although an improvement in protein‐uria levels (protein/creatinine ratio, from 171 to 126 mg/g; p=0.04) was observed soon after initiation.ConclusionsDual therapy with lamivudine plus darunavir boosted with ritonavir once daily is safe and effective as simplification strategy in HIV‐infected patients.
Folding and Purification of Insoluble (Inclusion Body) Proteins from Escherichia coli
Authors: Wingfield Paul T. , Palmer Ira , Liang Shu‐Mei 
Published: November 2014

AbstractHeterologous expression of recombinant proteins in E. coli often results in the formation of insoluble and inactive protein aggregates, commonly referred to as inclusion bodies. To obtain the native (i.e., correctly folded) and hence active form of the protein from such aggregates, four steps are usually followed: (1) the cells are lysed, (2) the cell wall and outer membrane components are removed, (3) the aggregates are solubilized (or extracted) with strong protein denaturants, and (4) the solubilized, denatured proteins are folded with concomitant oxidation of reduced cysteine residues into the correct disulfide bonds to obtain the native protein. This unit features three different approaches to the final step of protein folding and purification. In the first, guanidine·HCl is used as the denaturant, after which the solubilized protein is folded (before purification) in an “oxido‐shuffling” buffer system to increase the rate of protein oxidation. In the second, acetic acid is used to solubilize the protein, which is then partially purified by gel filtration before folding; the protein is then folded and oxidized by simple dialysis against water. Thirdly, folding and purification of a fusion protein using metal‐chelate affinity chromatography are described. © 2014 by John Wiley & Sons, Inc.