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.

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.

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.

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.

[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.

[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.

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.

[Determination of mold concentrations in homes and schools in Baden-Württemberg]. / Bestimmung der Schimmelpilzkonzentration in Wohnungen und Schulen Baden-Württembergs.

So far there have been rather few reliable and comparable data available on indoor pollution with mould. Following the publication of the Federal Environmental Agency and the Health Agency Baden-Württemberg which supports the assessment of mould pollution of indoor air, it seemed advisable to investigate as to how far these criteria can be used for the assessing the mould pollution in daily practice. The results of investigations of 130 homes and 117 classrooms in Baden-Württemberg. will be represented.

[Exposition and sensitisation to indoor allergens, house dust mite allergen and cat allergens]. / Exposition und Sensibilisierung gegenüber Milben- und Katzenallergenen bei Kindern in Baden-Württemberg.

The study examined the exposure to biological indoor air agents and their possible role for allergies and respiratory tract illnesses of children. It was conducted as a case control study (atopic vs non-atopic children) at the four surveillance public health departments in Baden-Württemberg in the winter season 1999/2000 and included 379 children of the fourth class. The concentrations of the house dust mite antigens Der F1, Der p1, and Der Gr2 as well as cat allergen Fel d1 were determined in the children's bedrooms on the ground and in the mattress. Specific IgE-antibodies against allergens from house dust, mites and cat were determined in the serum of the children. For mite allergens the following medians ( micro g/g) were estimated in floor dust: Der p1 = 0.6, Der f1 = 2.3, Gr2 = 0.1; in mattresses: Der p1 = 1.2, Der f1 = 3.4, Gr2 = 0.3. The median of Fel d1 in floor dust was 0.2 microg/g, in mattresses 0.1 microg/g. Sensitisation to dust mite allergen was found to be more prevalent than sensitisation to cat. The distribution of sensitisation among the cases and controls is different. Among the cases, more subjects were sensitised to dust mites (32.9 %) and cat (13.1 %). Among the controls, 17.1 % were sensitised to dust mites and 4.1 % to cat. The results showed no direct association between the prevalence of allergies or respiratory tract illnesses and the indoor concentrations of the allergens. Possible reasons for these findings are discussed.

[10 years of observation by public health offices in Baden-Württemberg--assessment of human biomonitoring for mercury due to dental amalgam fillings and other sources]. / 10 Jahre Beobachtungsgesundheitsämter in Baden-Württemberg--Beurteilung der Human-Biomonitoring-Untersuchungen bezüglich der Quecksilber-Belastung durch Amalgamfüllungen und andere Quellen.

Since 1992, in Baden-Württemberg, ten-year old children have been surveyed in the project "Sentinel Health Departments" to study their exposure to environmental pollutants and possible health effects. In the four study areas 1200 children have been investigated every year initially, since 1996 every second year. The data for mercury in body fluids are reported here. The decrease in the body burden of mercury as a result of the declining usage of dental amalgam fillings, was been verified. In 1992/93, of all the children who had been surveyed, the 95 percentile for the body burden of mercury was 3.1 microg/l and in 2000/01 1.35 microg/l. Also to be discussed is the reason why mercury-based cosmetic ointments seriously exceed the HBM-II-intervention-value. Because of using these ointments, concentrations of mercury in urine up to 1400 microg/l were found. A study within the project "Sentinel Health Departments" compared the concentrations of mercury in the urine of adults with those in blood and salvia. The results support the opinion that mercury in urine is appropriate for estimating the mercury uptake from dental amalgam fillings. It can be assumed that these results reflect the situation in the entire Federal Republic of Germany. The ten years' experience confirms that the concept of the "Sentinel Health Departments" is excellently suited to obtain data relevant for environmental health of children. Environmental health protection and the essential gathering of data for future health observation in Baden-Württemberg.

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.

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.

Sequential assessment of cell cycle S phase in flow cytometry: a non-isotopic method to measure lymphocyte activation in vitro.

Lymphocyte multiplication can be induced in vitro by mitogens or specific antigens, and is usually measured using isotopic methods involving tritiated thymidine. Cellular proliferation can also be analyzed by flow cytometry techniques based on cell cycle analysis through the measurement of DNA content. We applied this method to lymphocytes from 113 individuals, to evaluate lymphocyte proliferation after stimulation in vitro by a mitogen (phytohaemagglutinin, PHA) or a recall antigen (tetanus toxoid), using a kinetic approach with four points sequential measurements of the S and G2 phases over six days of culture. The proportion of cells in S phase after PHA stimulation was significantly higher than in controls overall and as early as on day three of the culture. Activation with a recall antigen significantly induced increasing S phase cell proportions up to day six. These data suggest that flow cytometric assessment of the S phase could be a useful alternative to isotopic methods measuring lymphocyte reactivity in vitro.

Autoantibodies against arterial antigens: characterization by ELISA and immunoblot analysis in the spontaneously hypertensive rat.

The presence of autoantibodies directed against arterial antigens in serum samples from spontaneously hypertensive rats and related controls that included Wistar-Kyoto and Sprague-dawley rats were assayed by enzyme-linked immunosorbent assay and immunoblotting technique. Circulating immunoglobulin G antibodies reactive against arterial antigen, as measured by enzyme-linked immunosorbent assay, could be detected in serum samples of 26 of 30 spontaneously hypertensive rats (87%) and 8 of 30 (27%) Wistar kyoto rats. These antibodies (Abs) were not detectable either by enzyme-linked immunosorbent assay or immunoblotting in sera from Sprague-dawley rats. The arterial antigen-reactive antibody was antigen specific, because the binding reactivity was absorbed by arterial antigen but not by fibroblasts or peripheral blood mononuclear cells. Immunoglobulin G arterial antigen-reactive antibody was significantly higher in adult spontaneously hypertensive rats with established hypertension, compared with young prehypertensive rats or normotensive wistar kyoto rats. Immunoblotting of spontaneously hypertensive rats sera revealed reactivity of arterial antigen-reactive antibody against arterial antigen ranging in size from 20 to 97 kDa. Sera from Wistar kyoto rats recognized arterial antigen ranging in size from 40 to 90 kDa. A significant correlation (p < 0.004) was found between adult spontaneously hypertensive rats with established hypertension and the presence of arterial antigen-reactive antibody reactivity against arterial antigen of 20, 69 and 97 kDa. Antibody directed against a 20 kDa arterial antigen was detected in both young prehypertensive rats and adult rats with established hypertension but not in Wistar kyoto or Sprague-dawley rats. Antibodies directed against both 69 and 97 kDa arterial antigens were detected only in spontaneously hypertensive rats sera. These data show that the pattern of arterial antigen-reactive antibody reactivity in sera of hypertensive rats in heterogeneous, and suggest that arterial antigen-reactive antibody directed against few arterial antigens may be involved in the pathogenesis of hypertension in the spontaneously hypertensive rat.

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.