Real-life rilpivirine resistance and potential emergence of an E138A-positive HIV strain in north-eastern France.

OBJECTIVES: To assess the prevalence of resistance to rilpivirine and mutations at position 138 in reverse transcriptase and to identify associated epidemiological and biological characteristics. METHODS: This retrospective study included 238 patients with available HIV-1 nucleotide sequences analysed at the Laboratory of Virology at the University Hospital of Nancy between January 2011 and June 2013. Resistance to non-nucleoside reverse transcriptase inhibitors (NNRTIs) was evaluated according to the ANRS algorithm (version 23) and correlated with clinico-epidemiological and therapeutic data. The virus strains were analysed by evaluating the distance and distribution of the phylogenetic tree (MEGAv5). RESULTS: Among previously treated patients (111/238, 46.6%), 68/111 (61.3%) had received NNRTIs; all were rilpivirine-naive. The prevalence of rilpivirine resistance in the whole cohort was 12.6% (30/238), and was 10.2% (13/127) and 15.3% (17/111) in naive and pre-treated patients, respectively. The E138A mutation was the most frequent mutation associated with resistance to rilpivirine (P < 0.0001). The prevalence of the E138A mutation tended to increase over time, from 3.6% (2/55) during the first half of 2011 to 9.3% (4/43) during the first half of 2013 (P = 0.0614). Seven viral strains from seven naive male patients positive for the E138A mutation appeared in the same cluster. CONCLUSIONS: In our cohort of patients, we observed significantly increased resistance to rilpivirine, mostly because of the E138A mutation, probably due to an E138A strain circulating in newly diagnosed men who have sex with men. Taken together, our results emphasize the need to investigate the prevalence of rilpivirine resistance-associated mutations in the coming years both in France and abroad.

[Impact of anti-HLA antibodies on outcomes of allogeneic stem cell transplantation: a report by the SFGM-TC]. / Impact des anticorps anti-HLA sur le devenir de l'allogreffe de cellules souches hématopoïétiques : un rapport par la SFGM-TC.

The role of anti-HLA antibodies in allogeneic stem cell transplantation setting is still unclear. In the attempt to harmonize clinical practices between different French transplantation centers, the French Society of Bone Marrow Transplantation and Cell Therapies (SFGM-TC) set up its fourth annual series of workshops which brought together practitioners from all of its member centers. These workshops took place in September 2013 in Lille. This article offers the recommendations of the group that considered the impact that have anti-HLA antibodies on outcomes in allogeneic stem cell transplantation.

HLA-B*51:79 is a novel allele associated with a conserved haplotype.

The B*51:79 allele displays a conserved haplotype association with HLA-A*68:01, C*01:02, DRB1*14:01 and DQB1*05:03.

[Discrepancy between two methods of D-dimers measurement: one case of human anti-mouse antibody interference]. / Incohérence entre deux méthodes de dosage des D-dimères: un cas d'interférence d'anticorps humain anti-souris.

Quantification of D-dimers is the major biometry step in the diagnostic of an episode of the venous thromboembolic disease. The measurement of D-dimers can be performed with ELISA or immunoturbidimetric methods suited to emergency, using a mouse monoclonal antibody as capture and/or revelation antibody. Therefore, the presence in patient's plasma of human antibody mouse (HAMA) that binds the mouse antiglobulin used in immunoassays can lead to false negative or false positive results. In a young woman presenting repetitive thoracic pain suggestive of a pulmonary embolism, a major discrepancy was found between one result of D-dimers above the cut-off with an immunoturbidimetric method (STA Liatest D-DI; Diagnostica Stago) and one result below the cut-off with a sandwich method (Vidas D-Dimer Exclusion; bioMérieux). HAMA, which is known to be responsible for this type of discrepancy, was detected in the patient serum. The false positive result probably impaired with the management of patient. Taking in charge the patient should take into account the possible presence of this antibody. Interference by heterophilic antibodies is not easily detected by the laboratory. Even if their frequency is low, it remains a difficult problem for the biologist. Suspicion generally arises from inconsistency between the clinical data and immunoassay results. A good communication between physician and biologist should avoid to providing false negative or positive results.

Temporal Trend and Time-Varying Effect of Preemptive Second Kidney Transplantation on Graft Survival: A 30-Year Single-Center Cohort Study.

BACKGROUND: There are discrepancies regarding the impact of preemptive 2nd kidney transplantation (PSKT) on graft survival. The present study aimed to determine whether the association between PSKT and outcome varies over time and whether this association is era dependent. METHODS: A total of 266 patients underwent SKT (244 non-PSKT, 22 PSKT) in our center from 1985 to 2015. Association between PSKT and graft survival (allograft failure from any cause including death) was assessed with the use of Cox models. RESULTS: During a median follow-up of 6.7 years, 116 events were recorded: 72 returns to dialysis and 44 deaths before return to dialysis. Survival curves diverged up to 5 years (5-year survivals: PSKT, 94.1 ± 5.7%; non-PSKT, 76.8 ± 2.9%) but they converged thereafter (12-year survivals: PSKT, 50.9 ± 15.2%; non-PSKT, 55.5 ± 3.9%). After adjustment for age and living-donor status, PSKT tended to be associated with better graft survival (hazard ratio [HR], 0.18; 95% confidence interval [CI], 0.02-1.27; P = .08) within the first 5 years of SKT but tended to be associated with worse outcome thereafter (HR, 2.36; 95% CI, 0.97-5.72; P = .06; P for interaction with time = .04). In addition, a significant interaction was identified between PSKT and SKT year (P for interaction = .04). In the multivariable model, the estimated HR for PSKT was 2.54 (95% CI, 0.88-7.35; P = .08) in 1990 as opposed to 0.16 (95% CI, 0.02-1.17; P = .07) in 2012. CONCLUSIONS: The effect of PSKT on graft survival varies over time and according to year of the procedure. Although the benefit observed within the first 5 years of SKT appears to fade over time, overall graft survival seemingly improved in more recent years.

Unusual association of myasthenia gravis and ulcerative colitis in a 14-year-old boy.

Ulcerative colitis (UC) is a chronic inflammatory condition. Its incidence is 0.8 per 100,000 children per year and median age at diagnosis is 14.1 years in the pediatric population. Extradigestive manifestations are frequent in inflammatory bowel disease (IBD), but neurologic disorders are rarely associated. We report the case of a 14-year-old boy who was diagnosed with UC and thereafter autoimmune hepatitis with primary sclerosing cholangitis. Two months later, he also developed myasthenia gravis, another autoimmune disease. Physicians should be aware of the possibility of an association between several autoimmune diseases in patients with IBD in order to improve disease outcomes.

Quality-of-care assessment. II. Outpatient medical care following hospital dismissal after myocardial infarction.

This study was undertaken to compare process and outcome methods of quality assessment of medical care in outpatient office practice. Follow-up care after hospitalization for first acute myocardial infarction was used as the model. One hundred fifty-two patients followed up for a minimum of 2 years comprised the study group. An expert committee of cardiologists and internists in community practice established the process criteria for satisfactory care and predicted outcomes of continuing disability and mortality. Using weighted process criteria and a weighted performance index permitted demonstration of a significant association between process items performed at the first posthospitalization visit and 2-year mortality. A significant association could not be demonstrated between later process of care and outcome at 2 years. The outcome assessment study disclosed that predicted disability and mortality rates compared closely with observed outcomes. However, this method for evaluating the quality of outpatient medical care is weakened because little information is available to provide the basis of prediction of satisfactory outcome rates in complicated cases. Although both the process and outcome methods of quality assessment have short comings, the latter method is recommended because satisfactory outcomes is the essential criterion of quality medical care. Moreover, when process items are not specified outcome assessment maintains the flexibility of individual physician practice. Refinement of satisfactory outcome prediction for common illnesses managed in office practice should be the goal for future studies.

Automated cell count in flow cytometry: a valuable tool to assess CD4 absolute levels in peripheral blood.

The enumeration of lymphocyte subsets in absolute counts has long relied on different methods applied separately to whole blood cell count, lymphocyte differential appreciation, and flow cytometric evaluation of lymphocyte subsets percentages. The development of multicolor labeling methods inflow cytometry now allows a more homogeneous appreciation of several cell subsets among gated lymphocytes. The use of internal calibrators, such as microbead suspensions, also permits a direct appreciation of subsets in absolute counts in a single-platform method. These methods were compared with a traditional multiplatform method of assessing absolute counts of lymphocyte subsets in a pilot study in which all manipulations were performed by 1 person and in a full-scale larger study performed in the normal working conditions of a hospital laboratory. Microspheres seem to be a reliable tool to perform absolute count enumeration inflow cytometry, but several precautions in the sample preparation and flow cytometric analysis are required.

Serum anti-dextran antibodies in IgA nephropathy.

Abnormally high humoral responses have been described in IgA nephropathy (IgAN), towards antigens commonly involved in infectious events or food intolerance. Here we report a comparative analysis of humoral responses to a common environmental antigen, dextran B 512, present both in nonpathogenic microorganisms and normal diet. Dextran-specific IgG, IgA and IgM were assayed using an ELISA method in serum samples from 121 patients with IgAN, 40 controls and 32 patients with biopsy-proven renal diseases different from IgAN. Among the latter, 17 were transplant recipients. Anti-dextran IgG antibodies were found significantly (p < 0.05) more frequently and at higher levels in IgAN patients than in patients with other renal diseases. Anti-dextran IgA were at significantly (p < 0.05) higher levels in IgAN patients than in controls or untransplanted NIgAN patients. However, similarly elevated levels of anti-dextran IgA were found in IgAN patients and transplanted patients with other renal diseases. These data confirm that IgA and IgG responses towards common antigens are elevated in IgAN, possibly as a consequence of the pathogenesis of this disease rather than because of renal impairment. Moreover, since the same degree of dysregulation was noted in the control group of transplanted patients whose initial diagnosis was not IgAN, these data favour the hypothesis of a global dysregulation of the IgA system in IgAN, yielding enhanced humoral immune responses to possible pathogens and diet antigens.

Surgical correction of an acquired vaginal stricture in a llama, using a carbon-dioxide laser.

An acquired vaginal stricture in a llama secondary to trauma from uterine torsion during gestation was repaired, using a carbon-dioxide laser. The procedure was accomplished by use of epidural anesthesia with the llama in a standing position. Repeated dilation of the vagina was required after surgery; however, the llama subsequently mated and conceived, and a male cria was delivered by cesarean section at the end of a full-term gestation.

Decrease of internal exposure to chlororganic compounds and heavy metals in children in Baden-Württemberg between 1996/1997 and 2008/2009.

From 1996/1997 to 2008/2009, blood and urine were sampled from 9- to 11-year-old pupils in the state of Baden-Württemberg, South-West Germany. In blood samples the chlororganics DDE, HCB, PCB-138, PCB-153 and PCB-180 were analysed by gas chromatography and ECD detection. PCDD/PCDF were measured in pooled blood samples using GC/MS. Lead concentrations in blood were quantified by graphite furnace atomic absorption spectrometry (AAS), mercury in urine by using cold vapour AAS. For all chlororganics and heavy metals a distinct decrease of the internal concentration could be shown within the 12-year investigation period. For DDE, the median decreased from 0.32 µg/l to 0.11 µg/l. The median of HCB fell from 0.19 µg/l to 0.07 µg/l. The median of the sum of the three PCB-congeners PCB-138, PCB-153 and PCB-180 decreased from 0.47 µg/l to 0.18 µg/l. The burden of PCDD/PCDF in pooled blood samples resulted in approximately half the concentration after the 12-year period. The internal concentrations of the chlororganics were distinctly higher in breastfed children than in non-breastfed children. In 2008/2009, these differences were strongest for PCB and DDE (2-fold), moderate for PCDD/PCDF (1.6-fold), and slight for HCB (1.2-fold). The median of lead concentrations in blood dropped from 23.6 µg/l to 15.9 µg/l. The median of mercury concentrations in urine decreased from 0.25 µg/l to a value below the limit of quantification of 0.2 µg/l; the 95th percentile was reduced from 3.1 µg/l to less than 0.2 µg/l. The decline of amalgam fillings in children during the investigation period had a strong influence on internal concentrations of mercury. The internal concentration of the persistent xenobiotics investigated here decreased to a low level not likely to be of concern for human health.

[Sensitization to airborne ragweed pollen--a cause of allergic respiratory diseases in Germany?]. / Sensibilisierung gegen Ambrosiapollen--Eine Ursache für allergische Atemwegserkrankungen in Deutschland?

BACKGROUND AND OBJECTIVE: Allergic skin and respiratory diseases show a high prevalence in most industrial countries. In addition, during the last years ragweed colonization has increased in Europe. Ambrosia pollen ( AMBROSIA ARTEMISIIFOLIA L. - common ragweed) are highly allergenic. Due to the late flowering time (august/September) of ragweed this can result in increasing health threats for allergic populations. This is of particular importance for those who already are sensitive to some grass or tree pollen. These individuals can then suffer from allergies during nearly the whole year. The present study examined the prevalence of sensitization to ragweed in German children and possible health implications. METHODS: Between 2004 and 2007 sera of 1323 10-years old children in Baden Württemberg were tested in-vitro for specific IgE-antibodies against common aeroallergens including ragweed pollen. RESULTS: Specific IgE-antibodies to extracts of common ragweed pollen were present in 10 - 17 % of the tested sera depending on the year of investigation. CONCLUSION: The determined specific IgE-antibodies may be the result of a direct sensitization to ragweed pollen or correspond to cross-reactivity to other plants of the asteraceae subfamily or some nutritional allergens. The detection of sensitization to ragweed pollen does not prove actual allergic disease. However, a ragweed derived allergy should be considered in the differential diagnosis when allergic symptoms are present in direct connection to the flowering-time of ragweed. Ragweed plants should be removed and the spread of the plant 'restricted', as experiences in other countries with already wide spreading show.

Human umbilical vein endothelial cells increase ex vivo expansion of human CD34(+) PBPC through IL-6 secretion.

BACKGROUND: Ex vivo expansion of hematopoietic stem cells (HSC) can help reduce cytopenia following transplantation, especially in NHL patients whose BM is deficient because of extensive chemotherapy. We have previously reported that human umbilical vein endothelial cells (HUVEC) can contribute to improved PBPC expansion when used in co-culture with CD34(+) cells. METHODS: We evaluated the roles of direct HUVEC CD34(+) contact and HUVEC-produced soluble factors. We cultured CD34(+) PBPC harvested from NHL patients in four different conditions: (1) liquid culture without HUVEC; (2) co-culture in contact with HUVEC; (3) co-culture with HUVEC but without direct contact; (4) liquid culture with HUVEC-conditioned medium (CM). Thrombopoietin (Tpo), Flk2Flt3 ligand (FL) and c-kit ligand (KL) with or without rhIL-6 were added to these four culture conditions. RESULTS AND DISCUSSION: Our results showed that HUVEC co-culture or addition of HUVEC-CM to Tpo, FL and KL (TFK) improved CD34(+) PBPC expansion compared with liquid culture, as determined by total viable nucleated cells (TNC), colony-forming cell assay (CFC) and week-6 cobblestone area-forming cells (Wk-6 CAFC) expansions. Non-contact culture led to similar PBPC expansion as contact co-culture; moreover, HUVEC-CM improved PBPC expansion. However, when rhIL-6 was added to HUVEC-CM with TFK, no significant difference was observed. Finally, high quantities of IL-6 were detected in HUVEC-CM and addition of anti-IL-6 Ab inhibited the positive effect of HUVEC on PBPC expansion. Our results thus suggest that HUVEC may improve PBPC expansion, at least through IL-6 secretion.

[Human biomonitoring investigations of organochlorine compounds -- PCB, DDE, HCB, beta- and gamma-HCH, PCDD/PCDF, Dioxin-like PCB's and polybrominated biphenyl ethers]. / Humanbiomonitoring-Untersuchungen von Organohalogenverbindungen -- PCB, DDE; HCB, beta- und gamma-HCH, PCDD/PCDF, koplanaren PCB sowie polybromierten Biphenylethern.

Although PCB and PCB-containing materials are not processed for a long time, PCB is under discussion again and again caused by the pollution of indoor environments. To objectify the discussion, the dates of the PCB-biomonitoring, the organochlorine-compounds (DDE, HCB, beta-/gamma -HCH, PCDD/PCDF) and the polybrominated biphenyl ethers concerning the investigations within the project "Sentinel Health Departments" in Baden-Wurttemberg are represented. Additionally results from children from Kazakhstan (Aral-Sea area) and from teachers which are working in PCB polluted schools as well as from a long term investigated test person are reported. Blood concentrations of the following compounds decreased from 1996/97 to 2002/03: the sum of the concentration of PCB 138,153 and 180 decreased from 0.46 microg/L to 0.20 microg/L, DDE from 0.32 microg/L to 0.17 microg/, HCB from 0.20 microg/L to 0.08 microg/L, beta-HCH below the level of detection, I-TEQ NATO to 4.8 pg/g blood fat, TEQ WHO (without PCB) to 5.5 pg/g blood fat, PCB 126 to 18,8,pg/g blood fat and PCB 169 to 12.8 pg/g blood fat. The influence of breast feeding and the gender on the level of the pollution is conspicious. No local correlations were found in Baden-Wurttemberg, but they were found in comparison with the results of Kazakhstan (Aral-Sea area). The difficulty to produce time series while the analyzing pollutants are more and more decreasing, as well as the change of the calculation base of the summation of parameters like I-TEQ NATO to TEQ WHO are discussed.