[Surgical urinary diversion in children from war and crisis areas-20-year experience report]. / Operative Harnableitung bei Kindern aus Kriegs- und Krisengebieten ­ 20-jähriger Erfahrungsbericht.

BACKGROUND: Because sick and injured children from war and crisis areas can often only be helped to a limited extent in their countries of origin, those children with the most complex diseases often receive medical treatment in Europe. MATERIALS AND METHODS: To evaluate the postoperative outcome of reconstructive interventions in children from war and crisis areas, an evaluation of the clinic's internal database between 1997 and 2017 was carried out. The operative indication, the surgical procedure, any revisions or conversions and the overall mortality were analyzed. RESULTS: During the above-mentioned period a total of 44 (male: 32, female: 12) children aged 3-14 years from Afghanistan, Angola, Central Asia and the Caucasus were treated. Indications for the reconstructive procedures were bladder exstrophy (n = 23), neurogenic bladder dysfunction (n = 9), penile partial amputation (n = 1) and traumatic bladder dysfunction (n = 11). In 26 children, a conduit was created, 11 children received a sigma rectum pouch and 1 child an ileocecal pouch, while 5 other children received bladder augmentation and 1 child an oral mucosal urethroplasty. In total, 7 revisions or conversions due to ureteral implantation stenoses and recurrent acidosis had to be performed. Five children died in their home country due to acidosis, sepsis, end stage renal failure, accident and social exclusion. CONCLUSION: In the long term 39 of the 44 (88.6%) children live socially integrated in their home country and are regularly monitored by local aid organizations. When choosing the reconstructive intervention, the local conditions must be considered.

A cross-sectional study of non-attendance among patients at a US hemophilia treatment center 2010-2014.

INTRODUCTION: Among patients with chronic disease, non-attendance at scheduled healthcare visits is associated with poor outcomes. The impact of non-attendance among patients with bleeding disorders is unknown. METHODS: Scheduling and medical record data over a 5-year period for all individuals with at least one scheduled appointment during 2010-2014 at a US Hemophilia Treatment Center (HTC) were analysed. Non-attendance rates were calculated as the number of non-attended visits divided by the number of years as a patient during the time period. Consistent non-attenders were patients who did not attend more than one scheduled appointment per person-year on average. Logistic regression determined characteristics associated with consistent non-attendance and Poisson regression estimated adjusted incidence rate ratios (aIRRs) describing associations between non-attendance and emergency department (ED) visits and hospitalizations. RESULTS: There were 8028 appointments scheduled for 950 individuals; 12% were not attended. Consistent non-attenders (n = 62; 7% of the HTC patient population) accounted for over one-third of non-attended appointments and over one-quarter of hospitalizations. Characteristics associated with consistent non-attendance included public health insurance and black race. Higher non-attendance rates were associated with more ED visits (aIRR 1.78; 95% CI: 1.37-2.30) and hospitalizations (aIRR 2.73; 95% CI: 2.18-3.42). Consistent non-attenders had more ED visits (aIRR 2.49; 95% CI: 1.56-3.96) and hospitalizations (aIRR 4.73; 95% CI: 2.96-7.57) compared with patients who never missed appointments. CONCLUSIONS: Frequent non-attendance identified a small but at-risk population. Interventions to improve disease management that target them may have an impact on health outcomes and healthcare utilization.

[Boys' health survey-between gender gap and information backlog]. / Wissenserhebung zum Thema Jungengesundheit ­ zwischen Geschlechterkluft und Nachholbedarf.

BACKGROUND: Early detection examinations take place from birth to the age of 6 years. The youth screening is a continuation of the screening of the "U-series" and should be carried out between the age of 12-15 and 16-17, respectively. Afterwards adolescent girls have good contact with a gynecologist, but adolescent boys usually do not have a medical contact person who they can trust in. MATERIALS AND METHODS: To evaluate the state of knowledge on boys' health, a 15-item comprehensive knowledge survey was conducted among ninth grade students at 7 secondary schools (Gymnasien) in North Rhine-Westphalia. The knowledge survey took place at three specified times (before, immediately after and approximately 3 months after adolescent sexual education classes). Only completed questionnaires were analyzed and evaluated in a gender-specific manner. RESULTS: Overall, 459 students participated from March-September 2017. Before sexual education instruction, about half of all questions were answered correctly by the students. Immediately after class, the proportion increased by a factor of 1.5 to a total of 79.24%. Then 2-3 months after the class, the percentage was 69.67%. Considering gender separately, this resulted in an increase of 15.32% for the female students and 16.99% for the male students. CONCLUSION: The knowledge survey reveals a need to catch up on facts on the subject of boys' health. Despite evidence of an increase in knowledge of both sexes after sexual education instruction, there is a gender gap. Hence, a preventive check-up especially for boys should be established and offered. Issues such as the prevention of sexually transmitted diseases, options for vaccination against human papillomavirus, etc. should be actively addressed.

Quality of life assessment in patients with metastatic colorectal cancer receiving maintenance therapy after first-line induction treatment: a preplanned analysis of the phase III AIO KRK 0207 trial.

BACKGROUND: First-line maintenance strategies are a current matter of debate in the management of mCRC. Their impact on patient's health-related quality of life (HRQOL) has not yet been evaluated. The objective of this study was to assess whether differences in HRQOL during any active maintenance treatment compared with no maintenance treatment exist. PATIENT AND METHODS: Eight hundred and thirty-seven patients were enrolled in the AIO KRK 0207 trial. Four hundred and seventy-two underwent randomization (after 24 weeks of induction treatment) into one of the maintenance arms: FP plus Bev (arm A), Bev alone (arm B), or no active treatment (arm C). HRQOL were assessed every 6 weeks during induction and maintenance treatment independent from treatment stop, delay, or modification, and also continued after progression, using the EORTC QLQ-C30, QLQ-CR29. The mean value of the global quality of life dimension (GHS/QoL) of the EORTC QLQ-C30, calculated as the average of all available time points after randomization was considered as pre-specified main endpoint. Additionally, EORTC QLQ-C30 response scores were analyzed. RESULTS: For HRQOL analysis, 413 patients were eligible (arm A: 136; arm B: 142, arm C: 135). Compliance rate with the HRQOL questionnaires was 95% at time of randomization and remained high during maintenance (98%, 99%, 97% and 97% at week 6, 12, 18 and 24). No significant differences between treatment arms in the mean GHS/QoL scores were observed at any time point. Also, rates of GHS/QoL score deterioration were similar (20.5%; 17.2% and 20.7% of patients), whereas a score improvement occurred in 36.1%; 43.8% and 42.1% (arms A, B and C). CONCLUSION: Continuation of an active maintenance treatment with FP/Bev after induction treatment was neither associated with a detrimental effect on GHS/QoL scores when compared with both, less active treatment with Bev alone or no active treatment. CLINICAL TRIALS NUMBER: NCT00973609 (ClinicalTrials.gov).

Capecitabine and bevacizumab with or without vinorelbine in first-line treatment of HER2/neu-negative metastatic or locally advanced breast cancer: final efficacy and safety data of the randomised, open-label superiority phase 3 CARIN trial.

The study was designed to evaluate efficacy and superiority of capecitabine/bevacizumab + vinorelbine (CAP/BEV/VIN) compared to CAP/BEV alone. Main purpose was to introduce a taxane-/anthracycline-free first-line treatment in advanced breast cancer (ABC), in order to avoid long-term toxicities. In this open-label, superiority, phase 3 trial, patients with HER2-negative ABC were randomized 1:1 to receive either oral CAP at 1000 mg/m(2) [twice daily, days 1-14, q3w] plus intravenous BEV at 15 mg/kg [day 1, q3w] (arm A) or in addition to this protocol intravenous VIN at 25 mg/m(2) [days 1 + 8, q3w] (arm B) until disease progression, unacceptable toxicity or withdrawal of consent. Between 26 February 2009 and 26 October 2012, we randomised 600 patients (arm A N = 300; arm B N = 300) from 57 German outpatient-centres and 2 university hospitals. Median progression-free survival (PFS) (primary endpoint) was not improved with VIN (CAP/BEV, 8.8 months; CAP/BEV/VIN, 9.6 months; HR 0.84 [95 % CI 0.70-1.01], P = 0.058). Median overall survival (OS) (secondary endpoint) was 25.1 and 27.2 months for CAP/BEV and CAP/BEV/VIN, respectively, average HR 0.85 [95 % CI 0.70-1.03], P = 0.104). The 1- and 2-year OS rates appeared to be similar (78.0 and 77.0 %; 53.0 and 54.0 %). Toxicity profiles were generally mild and manageable. Adverse events occurred more frequently in arm B. Regarding the balance between clinical efficacy (PFS, OS) and toxicity, the CAP/BEV combination provides a favourable treatment option in first-line ABC avoiding taxane- and/or anthracycline-induced long-term toxicity. Superiority of CAP/BEV/VIN was not met, and side effects were even enhanced. Nevertheless, no safety issues occurred.

Water quality assessment of the Sinos River - RS, Brazil.

Worldwide environmental pollution is increasing at the same rate as social and economic development. This growth, however, is disorganized and leads to increased degradation of water resources. Water, which was once considered inexhaustible, has become the focus of environmental concerns because it is essential for life and for many production processes. This article describes monitoring of the water quality at three points along the Sinos River (RS, Brazil), one in each of the upper, middle and lower stretches. The points were sampled in 2013 and again in 2014. The water samples were analyzed to determine the following physical and chemical parameters plus genotoxicity to fish: metals (Cr, Fe, Al), chemical oxygen demand, biochemical oxygen demand, chlorides, conductivity, total suspended solids, total phosphorous, total and fecal coliforms, pH, dissolved oxygen, turbidity, total Kjeldahl nitrogen nitrate and ammoniacal nitrogen. Genotoxicity was tested by exposing individuals of the species Astyanax jacuhiensis to water samples and then comparing them with a control group exposed to water from the public water supply. The results confirmed the presence of substances with genotoxic potential at the sample points located in the middle and lower stretches of the river. The results for samples from the upper stretch, at P1, did not exhibit differences in relation to the control group. The physical and chemical analyses did not detect reductions in water quality in the lower stretch, as had been expected in view of the large volumes of domestic and industrial effluents discharged into this part of the river.

Water quality assessment of the Sinos River – RS, Brazil / Qualidade da água do Rio dos Sinos – RS, Brasil

Worldwide environmental pollution is increasing at the same rate as social and economic development. This growth, however, is disorganized and leads to increased degradation of water resources. Water, which was once considered inexhaustible, has become the focus of environmental concerns because it is essential for life and for many production processes. This article describes monitoring of the water quality at three points along the Sinos River (RS, Brazil), one in each of the upper, middle and lower stretches. The points were sampled in 2013 and again in 2014. The water samples were analyzed to determine the following physical and chemical parameters plus genotoxicity to fish: metals (Cr, Fe, Al), chemical oxygen demand, biochemical oxygen demand, chlorides, conductivity, total suspended solids, total phosphorous, total and fecal coliforms, pH, dissolved oxygen, turbidity, total Kjeldahl nitrogen nitrate and ammoniacal nitrogen. Genotoxicity was tested by exposing individuals of the species Astyanax jacuhiensis to water samples and then comparing them with a control group exposed to water from the public water supply. The results confirmed the presence of substances with genotoxic potential at the sample points located in the middle and lower stretches of the river. The results for samples from the upper stretch, at P1, did not exhibit differences in relation to the control group. The physical and chemical analyses did not detect reductions in water quality in the lower stretch, as had been expected in view of the large volumes of domestic and industrial effluents discharged into this part of the river.

Resumo A poluição ambiental aumenta mundialmente, no mesmo ritmo que o desenvolvimento social e econômico. Este crescimento é, muitas vezes, desorganizado e proporciona um aumento da degradação dos recursos hídricos. Água, que já foi considerado inesgotável, tornou-se o foco das preocupações ambientais, pois é essencial para a vida e para muitos processos de produção. Este artigo aborda o monitoramento da qualidade da água, em três pontos do Rio dos Sinos, compreendendo o trecho superior, médio e inferior. Os pontos foram monitorados entre 2013 e 2014. As amostras de água foram analisadas para determinar os parâmetros físico-químicos complementarmente a genotoxicidade em peixes: metais (Cr, Fe, Al), demanda química de oxigenio, demanda bioquímica de oxigênio, cloretos, condutividade, sólidos suspensos totais, fósforo total, coliformes totais e fecais, pH, oxigênio dissolvido, turbidez, nitrogênio total Kjeldahl, nitrato e nitrogênio amoniacal. A avaliação de genotoxicidade foi conduzida pela exposição à água de diferentes pontos do rio de indivíduos da espécie Astyanax jacuhiensis. O grupo controle foi exposto à água de abastecimento público. Os resultados confirmam a presença de substâncias com potencial genotóxico nos pontos localizados nos trechos médio e inferior. No trecho superior, ponto 1, não houve diferenças em relação ao grupo controle. A diminuição da qualidade da água do trecho inferior quando comparada com o superior também foi confirmada através das análises físico-químicas, as quais indicaram uma redução na qualidade da água nos trechos inferiores, como o esperado devido ao grande escoamento de esgotos domésticos e industriais neste trecho do rio.

Water quality assessment of the Sinos River RS, Brazil

Worldwide environmental pollution is increasing at the same rate as social and economic development. This growth, however, is disorganized and leads to increased degradation of water resources. Water, which was once considered inexhaustible, has become the focus of environmental concerns because it is essential for life and for many production processes. This article describes monitoring of the water quality at three points along the Sinos River (RS, Brazil), one in each of the upper, middle and lower stretches. The points were sampled in 2013 and again in 2014. The water samples were analyzed to determine the following physical and chemical parameters plus genotoxicity to fish: metals (Cr, Fe, Al), chemical oxygen demand, biochemical oxygen demand, chlorides, conductivity, total suspended solids, total phosphorous, total and fecal coliforms, pH, dissolved oxygen, turbidity, total Kjeldahl nitrogen nitrate and ammoniacal nitrogen. Genotoxicity was tested by exposing individuals of the species Astyanax jacuhiensis to water samples and then comparing them with a control group exposed to water from the public water supply. The results confirmed the presence of substances with genotoxic potential at the sample points located in the middle and lower stretches of the river. The results for samples from the upper stretch, at P1, did not exhibit differences in relation to the control group. The physical and chemical analyses did not detect reductions in water quality in the lower stretch, as had been expected in view of the large volumes of domestic and industrial effluents discharged into this part of the river.

Resumo A poluição ambiental aumenta mundialmente, no mesmo ritmo que o desenvolvimento social e econômico. Este crescimento é, muitas vezes, desorganizado e proporciona um aumento da degradação dos recursos hídricos. Água, que já foi considerado inesgotável, tornou-se o foco das preocupações ambientais, pois é essencial para a vida e para muitos processos de produção. Este artigo aborda o monitoramento da qualidade da água, em três pontos do Rio dos Sinos, compreendendo o trecho superior, médio e inferior. Os pontos foram monitorados entre 2013 e 2014. As amostras de água foram analisadas para determinar os parâmetros físico-químicos complementarmente a genotoxicidade em peixes: metais (Cr, Fe, Al), demanda química de oxigenio, demanda bioquímica de oxigênio, cloretos, condutividade, sólidos suspensos totais, fósforo total, coliformes totais e fecais, pH, oxigênio dissolvido, turbidez, nitrogênio total Kjeldahl, nitrato e nitrogênio amoniacal. A avaliação de genotoxicidade foi conduzida pela exposição à água de diferentes pontos do rio de indivíduos da espécie Astyanax jacuhiensis. O grupo controle foi exposto à água de abastecimento público. Os resultados confirmam a presença de substâncias com potencial genotóxico nos pontos localizados nos trechos médio e inferior. No trecho superior, ponto 1, não houve diferenças em relação ao grupo controle. A diminuição da qualidade da água do trecho inferior quando comparada com o superior também foi confirmada através das análises físico-químicas, as quais indicaram uma redução na qualidade da água nos trechos inferiores, como o esperado devido ao grande escoamento de esgotos domésticos e industriais neste trecho do rio.

Microcantilever sensors coated with doped polyaniline for the detection of water vapor.

In the present work, PANI (polyaniline) emeraldine salt (doped) and base (dedoped) were used as the sensitive layer of a silicon microcantilever, and the mechanical response (deflection) of the bimaterial (coated microcantilever) was investigated under the influence of humidity. PANI in the emeraldine base oxidation state was obtained by interfacial synthesis and was deposited on the microcantilever surface by spin-coating (dedoped). Next, the conducting polymer was doped with 1 M HCl (hydrochloric acid). A four-quadrant AFM head with an integrated laser and a position-sensitive detector (AFM Veeco Dimension V) was used to measure the optical deflection of the coated microcantilever. The deflection of the coated (doped and undoped PANI) and uncoated microcantilever was measured under different humidities (in triplicate) at room pressure and temperature in a closed chamber to evaluate the sensor's sensitivity. The relative humidity (RH) in the chamber was varied from 20% to 70% using dry nitrogen as a carrier gas, which was passed through a bubbler containing water to generate humidity. The results showed that microcantilevers coated with sensitive layers of doped and undoped PANI films were sensitive (12,717 ± 6% and 6,939 ± 8%, respectively) and provided good repeatability (98.6 ± 0.015% and 99 ± 0.01%, respectively) after several cycles of exposure to RH. The microcantilever sensor without a PANI coating (uncoated) was not sensitive to humidity. The strong effect of doping on the sensitivity of the sensor was attributed to an increased adsorption of water molecules dissociated at imine nitrogen centers, which improves the performance of the coated microcantilever sensor. Moreover, microcantilever sensors coated with a sensitive layer provided good results in several cycles of exposure to RH (%).

Microcantilever sensors coated with a sensitive polyaniline layer for detecting volatile organic compounds.

This paper describes a silicon cantilever sensor coated with a conducting polymer layer. The mechanical response (deflection) of the bimaterial (the coated microcantilever) was investigated under the influence of several volatile compounds-methanol, ethanol, acetone, propanol, dichloroethane, toluene and benzene. The variations in the deflection of the coated and uncoated microcantilevers when exposed to volatile organic compounds were evaluated, and the results indicated that the highest sensitivity was obtained with the coated microcantilever and methanol. The uncoated microcantilever was not sensitive to the volatile organic compounds. An increase in the concentration of the volatile organic compound resulted in higher deflections of the microcantilever sensor. The sensor responses were reversible, sensible, rapid and proportional to the volatile concentration.

Calagem e adubação fosfatada favorecem o crescimento do capim-limão, Cymbopogon citratus (DC) Stapf / Liming and phosphate fertilization promote lemongrass, Cymbopogon citratus (DC) Stapf. growth

O capim-limão, Cymbopogon citratus (DC) Stapf., é uma planta utilizada para fins medicinais e aromáticos, porém pouco se conhece sobre as exigências nutricionais. O objetivo do presente trabalho foi avaliar os efeitos da elevação do pH do solo e da adubação com fósforo (P) na disponibilidade de nutrientes e no crescimento inicial do capim-limão. O experimento foi conduzido em Lages, SC, de abril a dezembro de 2008, em casa de vegetação. Foi utilizado um esquema fatorial, incluindo quatro níveis de pH (4,1; 5,5; 6,0 e 6,5) e três doses de P (0, 50 e 100 mg kg-1 de solo), com quatro repetições (correspondendo a um vaso com duas plantas). A correção do pH aumentou o Ca e Mg e diminuiu o Al, Mn, Fe, Cu e Zn no solo. A aplicação de 100 mg kg-1 de P elevou em mais de 250% a concentração deste nutriente no solo, independente do pH. A aplicação de P e, principalmente, a elevação do pH do solo incrementaram todos os atributos avaliados nas plantas de capim limão (massa seca da parte aérea e do sistema radicular, número de folhas e altura de plantas), sendo que a interação entre pH e dose de P somente foi significativa para o número de folhas. O maior crescimento das plantas ocorreu no tratamento com pH 6,5, associado à adição de 100 mg kg-1 de P.

Lemongrass, Cymbopogon citratus (DC) Stapf., is a plant with medicinal and aromatic uses. However, little is known about its nutrition requirements. This work was carried out to investigate the effects of pH and phosphorus levels on nutrients availability and the initial growth of lemongrass. The experiment was carried out in Lages (SC, Brazil) from April tol December 2008, in greenhouse conditions. The experiment followed a factorial design, with four levels of pH (4.1, 5.5, 6.0, and 6.5), three doses of phosphorus (0, 50, and 100 mg kg-1 of soil), and four replicates (each replicate corresponding to a pot with two plants). The correction of pH increased Ca and Mg, and reduced Al, Mn, Fe, Cu, and Zn in the soil. The addition of 100 mg kg-1 P increased by more than 250% the P in the soil, irrespective of pH. The addition of P and the correction of pH, specially the last one, increased all growth attributes of lemongrass (dry matter of shoot and root parts, number of leaves, and plant height), with a significant interaction between pH and the dose of P only for the number of leaves. The highest plant growth was achieved by correcting the soil pH to 6.5, with the addition of 100 mg kg-1 P.

Desempenho e composição dos filés de jundiás (Rhamdia quelen) submetidos a diferentes dietas na fase de recria / Performance and fillet composition of jundiá (Rhamdia quelen) submitted to different diets in the rearing

Avaliaram-se o crescimento e a composição dos filés de juvenis de jundiá após 90 dias de alimentação com dietas contendo diferentes fontes protéicas. Foram utilizados 540 peixes, peso inicial=15,0±0,62g; comprimento inicial=11,98±0,35cm, distribuídos ao acaso em 18 tanques (30 peixes/tanque) com sistema de recirculação de água. Foram testadas seis dietas: 1 - farinha de carne e ossos + levedura (CL), 2 - farelo de soja + levedura (SL), 3 - farelo de soja (S), 4 - farinha de carne e ossos + farelo de soja (CS), 5 - farinha de peixe + levedura (PL) e 6 - farinha de peixe + farelo de soja (PS). Avaliaram-se: peso, comprimento total, taxa de crescimento específico, fator de condição, sobrevivência, conversão alimentar aparente, taxa de eficiência protéica e consumo diário. Também foram analisadas as composições do filé - umidade, proteína, lipídios, cinzas -, as taxas de deposição de proteína e gordura, além dos rendimentos de filé e carcaça. Maiores pesos, 111,5 e 108,6g, comprimento total, 22,6 e 21,6cm, e taxa de crescimento específico, 2,1 e 2,2 por cento/dia, foram observados nos peixes alimentados com as dietas CS e PS. Os peixes alimentados com CS e PS também apresentaram maior deposição de proteína, 69,8 e 71,5mg/dia, e lipídios, 77,2 e 78,3mg/dia. A sobrevivência foi menor na dieta PS, 92,9 por cento. Os maiores rendimentos de carcaça foram observados nos peixes alimentados com PS e PL, 82,9 e 82,7 por cento, e o menor rendimento de filé ocorreu na dieta CL, 34,1 por cento.

The growth and fillet composition of jundiá juveniles after 90 days being fed with diets containing different protein sources were evaluated. Five hundred and forty fish (initial weight: 15.00±0.62g; initial length: 11.98±0.35cm) were randomly distributed in 18 tanks (30 fish/tank), in a water re-use system. Six diets were tested: 1- meat and bone meal + yeast (CL), 2- soybean meal + yeast (SL), 3- soybean meal (S), 4- meat and bone meal + soybean meal (CS), 5- fish meal + yeast (PL,) and 6- fish meal + soybean meal (PS). The following parameters were evaluated: weight, total length, specific growth rate, condition factor, survival, feed conversion ratio, protein efficiency ratio, and daily consumption. Fillet composition (moisture, protein, lipid, and ash), protein and lipid depositions ratios, carcass and fillet yields were also determined. The highest values of weight (111.5 and 108.6g), total length (22.6 and 21.6cm), and specific growth rate (2.1 and 2.2 percent/day), were observed in fish fed CS and PS diets. Fish fed CS and PS diets also showed the highest protein (69.8 and 71.5mg/day), and lipid depositions (77.2 and 78.3mg/day). Survival was smaller in fish fed diet PS (92.86 percent). The highest values of carcass yield were observed in fish fed PL and PS diets (82.9 and 82.7 percent), and the lower fillet yield was observed in CL diet (34.1 percent).

[Twiddler's syndrome in a left pectoral implanted defibrillator]. / Twiddler-Syndrom bei einem links pektoral implantierten Defibrillator.

Twiddler's syndrome describes a rare complication following implantation of a pacemaker/defibrillator and is characterized by spontaneous, repeated rotation of the generator in the subpectoral pocket. We describe the case of a 78-year-old man who presented with an exit block 7 months after defibrillator implantation. As cause, a 15-fold rotation of the defibrillator's generator around its own axis could be verified, which entailed dislocation of the defibrillator's electrode. Safe fixation of the electrode and generator in the subpectoral pocket is of paramount importance, particularly in patients with loose subcutaneous fatty tissue, to prevent this rare complication.

T cell receptor excision circle (TREC) content following maximum HIV suppression is equivalent in HIV-infected and HIV-uninfected individuals.

BACKGROUND: The adult human thymus contributes to de novo T cell synthesis; such synthesis can be assessed by analyzing T cell receptor excision circles (TREC). METHODS: TREC levels were measured in total peripheral blood mononuclear cells (PBMC) and CD4- and CD8-enriched cells of 29 HIV-positive patients with maximal viral suppression. The expression of CD45RA+CD45RO-, CD45RA+CD62L+, CD45RO-CD27+CD95low and HLA-DR+CD38+ was assessed using three-color flow cytometric analysis of whole blood. Thymic index score was based on computed tomographic scans of the thymus. The relationship of TREC with thymic index and the expression of the naive phenotypes was evaluated. RESULTS: TREC expression was not statistically different in these HIV-positive patients from that in age-matched HIV-negative controls. Among HIV-positive patients with CD4 cell count of > 500 x 10(6) cells/l after antiretroviral therapy (n = 15), PBMC TREC levels correlated with the expression of CD45RA+CD45RO- and CD45RA+CD62L+ naive phenotypes, and inversely correlated with the expression of HLA-DR+CD38+. The change between pre- and post-therapy CD4 cell counts for these 15 patients significantly correlated with both thymic index and expression of the CD45RA+CD45RO- phenotype. CONCLUSIONS: The finding that TREC expression was equivalent between HIV-positive patients after therapy and HIV-negative donors suggests that there is no reduction in thymic output among HIV-positive individuals after therapy. Given that TREC is inversely correlated with HLA-DR/CD38 expression, its analysis in studies of thymopoiesis should be evaluated in the context of maximum viral suppression to reduce HIV-mediated immune activation and/or by normalizing for cell turnover.

Evaluation of thymopoiesis using T cell receptor excision circles (TRECs): differential correlation between adult and pediatric TRECs and naïve phenotypes.

To determine whether the thymus is still functional despite age-related involution, we measured a biomarker for thymopoiesis known as the T cell receptor excision circle (TREC) from peripheral blood mononuclear cells (PBMCs) of 148 healthy children and from PBMCs, CD4(+), and CD8(+) cells of 32, 30, and 50 healthy adults, respectively. We demonstrate that during the first 5 years of life, thymic output is decreased (P 0.002) but not dramatically (r = -0. 282). Among adults aged 23-58, thymic output was inversely correlated with age, as measured from PBMCs (r = -0.628, P < 0.0005), CD4(+) (r = -0.530, P 0.003), and CD8(+) fractions (r = -0.385, P 0. 006). A strong correlation existed between pediatric PBMC TRECs and the expression of three naïve phenotypic markers (CD45RA(+)CD45RO(-), CD45RA(+)CD62L(+), and CD45RO(-)CD27(+)CD95(low)). Adult PBMC TRECs correlated only with the expression of CD45RA(+)CD45RO(-) (r = 0.459, P 0.012). Our data suggest that in adults CD45RA(+)CD45RO(-) may be enriched for TRECs and add to a growing body of evidence illustrating intact thymic function in adulthood.

Detection of T cell receptor circles (TRECs) as biomarkers for de novo T cell synthesis using a quantitative polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA).

Currently, phenotypic markers that distinguish between recent thymic emigrants/de novo T cells and the rest of the peripheral T cell pool are lacking. This distinction is critical in studies aimed at evaluating immune reconstitution following intensive chemotherapy, in immunodeficiency-related therapies, or in the elucidation of the kinetics of thymic function. During V(D)J T cell receptor rearrangement, DNA extrachromosomal excision products are generated. These products, known as T cell receptor excision circles (TRECs), are not replicated during mitosis and are thus diluted with each round of cell division. Therefore, TRECs can be used as an indicator of recent thymic emigrants. Thus far, quantitative competitive-polymerase chain reaction (QC-PCR) and real time PCR were used to measure TREC levels. However, QC-PCR relies on radioactivity, is cumbersome when processing many samples at once and the cost of real time PCR does not make it a viable option for many laboratories. We describe here the development of a quantitative PCR-ELISA method for the measurement of coding joint TRECs generated from ValphaJalpha recombination. Our assay is ultra sensitive, relies on biotin labeling rather than radioactivity, is based on a 96-well format making multiple process sampling relatively easy, and is cost effective. Using this PCR-ELISA method, we evaluated thymic output among 22 normal subjects, ranging in age from 22-53 years, and among HIV-infected individuals following highly active antiretroviral therapy (HAART). We demonstrate that an inverse relationship exists between TREC levels and aging in normal individuals and that, among some HIV patients, HAART treatment leads to enhanced thymic output. Our assay has direct relevance in projects examining normal and abnormal thymic function and in immune reconstitution studies.

Sequence analysis of an amphioxus cosmid containing a gene homologous to members of the aldo-keto reductase gene superfamily.

To gain an insight into vertebrate genome evolution, we have analysed the organization of an approximately 40-kb genomic clone of an amphioxus (Branchiostoma floridae) cosmid library. Amphioxus is considered as being the last non-vertebrate relative to vertebrates. Sequencing and analysis of the above clone using three different exon prediction programs (Grail, GenScan, Mzef) have led to the identification of a gene of the aldo-keto reductase family as well as further exons that gave a significant database match to known genes.

The genomic organization of NKG2C, E, F, and D receptor genes in the human natural killer gene complex.

Interactions of natural killer cell receptors with their cognate ligands play a major role in regulating NK cell function. The NKG2 gene family encodes several highly similar proteins, which are known to form heterodimers with the CD94 receptor. These dimers play a role in the inhibition as well as the activation of NK cells. We have analyzed the gene structures of the NKG2C, D, E, and F genes, and determined their genomic organization. Restriction mapping and sequencing revealed the four genes to be closely linked to one another, and of the same transcriptional orientation. An exon duplication within the NKG2C and E genes was identified, although the duplicated version of this exon has not yet been found in mRNA sequences. The NKG2C, E, and F genes, despite being highly similar, are variable at their 3' ends. We show that NKG2C consists of six exons, whereas NKG2E has seven, and the splice acceptor site for the seventh exon occurs in an Alu repeat. NKG2F consists of only four exons and part of exon IV is in some cases spliced to the 5' end of the NKG2D transcript. NKG2D has only a low similarity to the other NKG2 genes.

Effects of age and estrogen status on the skeletal IGF regulatory system. Studies with human marrow.

Human marrow was obtained as material discarded during total hip replacement and was established in culture with phenol red-free alpha-MEM with 10% fetal bovine serum (FBS) and antibiotics. Insulin-like growth factor I (IGF-I) and its binding proteins were secreted by human marrow cells, in amounts that increased with time in culture. Western ligand blotting showed that insulin-like growth factor binding protein-3 (IGFBP-3) accounted for the majority (approximately 75%) of the secreted binding proteins. Evidence for marrow secretion of BP-3 protease was found by electrophoretic analysis of mixtures of radiolabeled IGFBP-3 and marrow-conditioned media. The amount of constitutive secretion of IGFBP-3 increased with age of the subject (r = 0.97, p = 0.0058). A notable exception was marrow from a postmenopausal women on estrogen replacement therapy (ERT) at the time of surgery; her marrow secreted 89.3 ng/mL after 14 d in vitro, only 38% of the IGFBP-3 that was secreted by cultures from two age-matched peers (208.8 and 285.2 ng/mL). This in vivo effect of estrogen was matched by an in vitro experiment in which 10(-8) M 17-beta estradiol suppressed IGFBP-3 to 60% of the constitutive level. In all cultures of marrow from postmenopausal women, IL-1 beta suppressed IGFBP-3 secretion to either undetectable levels or levels between 11% and 35% of control. Thus, human bone marrow cultures demonstrate components of the skeletal IGF regulatory system: IGF-I, IGF-binding proteins, and evidence of IGFBP-3 proteolysis. These results provide evidence of regulation by both systemic (age, estrogen status) and cytokine (IL-1 beta) factors.