Cell disposition of raltegravir and newer antiretrovirals in HIV-infected patients: high inter-individual variability in raltegravir cellular penetration.

Objectives The site of pharmacological activity of raltegravir is intracellular. Our aim was to determine the extent of raltegravir cellular penetration and whether raltegravir total plasma concentration (C(tot)) predicts cellular concentration (C(cell)). Methods Open-label, prospective, pharmacokinetic study on HIV-infected patients on a stable raltegravir-containing regimen. Plasma and peripheral blood mononuclear cells were simultaneously collected during a 12 h dosing interval after drug intake. C(tot) and C(cell) of raltegravir, darunavir, etravirine, maraviroc and ritonavir were measured by liquid chromatography coupled to tandem mass spectrometry after protein precipitation. Longitudinal mixed effects analysis was applied to the C(cell)/C(tot) ratio. Results Ten HIV-infected patients were included. The geometric mean (GM) raltegravir total plasma maximum concentration (C(max)), minimum concentration (C(min)) and area under the time-concentration curve from 0-12 h (AUC(0-12)) were 1068 ng/mL, 51.1 ng/mL and 4171 ng·h/mL, respectively. GM raltegravir cellular C(max), C(min) and AUC(0-12) were 27.5 ng/mL, 2.9 ng/mL and 165 ng·h/mL, respectively. Raltegravir C(cell) corresponded to 5.3% of C(tot) measured simultaneously. Both concentrations fluctuate in parallel, with C(cell)/C(tot) ratios remaining fairly constant for each patient without a significant time-related trend over the dosing interval. The AUC(cell)/AUC(tot) GM ratios for raltegravir, darunavir and etravirine were 0.039, 0.14 and 1.55, respectively. Conclusions Raltegravir C(cell) correlated with C(tot) (r = 0.86). Raltegravir penetration into cells is low overall (∼5% of plasma levels), with distinct raltegravir cellular penetration varying by as much as 15-fold between patients. The importance of this finding in the context of development of resistance to integrase inhibitors needs to be further investigated.

Gain-of-function haplotype in the epithelial calcium channel TRPV6 is a risk factor for renal calcium stone formation.

The rate-limiting step of dietary calcium absorption in the intestine requires the brush border calcium entry channel TRPV6. The TRPV6 gene was completely sequenced in 170 renal calcium stone patients. The frequency of an ancestral TRPV6 haplotype consisting of three non-synonymous polymorphisms (C157R, M378V, M681T) was significantly higher (P = 0.039) in calcium stone formers (8.4%; derived = 502, ancestral = 46) compared to non-stone-forming individuals (5.4%; derived = 645, ancestral = 37). Mineral metabolism was investigated on four different calcium regimens: (i) free-choice diet, (ii) low calcium diet, (iii) fasting and (iv) after a 1 g oral calcium load. When patients homozygous for the derived haplotype were compared with heterozygous patients, no differences were found with respect to the plasma concentrations of 1,25-vitamin D, PTH and calcium, and the urinary excretion of calcium. In one stone-forming patient, the ancestral haplotype was found to be homozygous. This patient had absorptive hypercalciuria. We therefore expressed the ancestral protein (157R+378V+681T) in Xenopus oocytes and found a significantly enhanced calcium permeability when tested by a (45)Ca(2+) uptake assay (7.11 +/- 1.93 versus 3.61 +/- 1.01 pmol/min/oocyte for ancestral versus derived haplotype, P < 0.01). These results suggest that the ancestral gain-of-function haplotype in TRPV6 plays a role in calcium stone formation in certain forms of absorptive hypercalciuria.

Response-to-comments about: "Is it really the method for carbon dioxide determination in human postmortem cardiac gas samples using Headspace-Gas Chromatography-Mass Spectrometry valid?" from T. Saffaj and B. Ihssane

Saffaj et al. recently criticized our method of monitoring carbon dioxide in human postmortem cardiac gas samples using Headspace-Gas Chromatography-Mass Spectrometry. According to the authors, their demonstration, based on the latest SFSTP guidelines (established after 2007 [1,2]) fitted for the validation of drug monitoring bioanalytical methods, has put in evidence potential errors. However, our validation approach was built using SFSTP guidelines established before 2007 [3-6]. We justify the use of these guidelines because of the post-mortem context of the study (and not clinical) and the gaseous state of the sample (and not solid or liquid). Using these guidelines, our validation remains correct.

Interferon-Induced Gene Expression Is a Stronger Predictor of Treatment Response Than IL28B Genotype in Patients With Hepatitis C.

BACKGROUND & AIMS: The host immune response during the chronic phase of hepatitis C virus infection varies among individuals; some patients have a no interferon (IFN) response in the liver, whereas others have full activation IFN-stimulated genes (ISGs). Preactivation of this endogenous IFN system is associated with nonresponse to pegylated IFN-α (pegIFN-α) and ribavirin. Genome-wide association studies have associated allelic variants near the IL28B (IFNλ3) gene with treatment response. We investigated whether IL28B genotype determines the constitutive expression of ISGs in the liver and compared the abilities of ISG levels and IL28B genotype to predict treatment outcome. METHODS: We genotyped 109 patients with chronic hepatitis C for IL28B allelic variants and quantified the hepatic expression of ISGs and of IL28B. Decision tree ensembles, in the form of a random forest classifier, were used to calculate the relative predictive power of these different variables in a multivariate analysis. RESULTS: The minor IL28B allele was significantly associated with increased expression of ISG. However, stratification of the patients according to treatment response revealed increased ISG expression in nonresponders, irrespective of IL28B genotype. Multivariate analysis of ISG expression, IL28B genotype, and several other factors associated with response to therapy identified ISG expression as the best predictor of treatment response. CONCLUSIONS: IL28B genotype and hepatic expression of ISGs are independent predictors of response to treatment with pegIFN-α and ribavirin in patients with chronic hepatitis C. The most accurate prediction of response was obtained with a 4-gene classifier comprising IFI27, ISG15, RSAD2, and HTATIP2.

Increased endometrial placenta growth factor (PLGF) gene expression in women with successful implantation.

Objective: To analyze the vascularization of the endometrium via hysteroscopy and to assess its correlation with angiogenic factor gene expression and embryo implantation rate.Design: Cross-sectional study.Setting: Public university hospital.Patient(s): Patients undergoing hysteroscopy for supposed infertility.Intervention(s): Endometrial quality assessment according to Sakumoto-Masamoto, performed in the early secretory phase of the cycle. Collection of an endometrial tissue biopsy.Main Outcome Measure(s): RNA extraction, reverse transcription, and determination of gene expression of angiogenesis- and implantation-relevant factors using quantitative polymerase chain reaction. Retrieval of pregnancy information from the medical records.Result(s): Good quantity/quality RNA with infertility history was obtained from 63 participating women. Those with a "good" endometrium and subsequent pregnancy showed increased gene expression for placenta growth factor when compared with patients with a "bad" endometrium and who did not succeed with pregnancy to date. Nonpregnant women with a "good" endometrium presented an intermediate result. No significant differences were observed for several other genes tested, but trends in the same direction were observed.Conclusion(s): This study demonstrates for the first time that endometrial PLGF expression corresponds to the hysteroscopic appearance of the endometrium, and therefore has potential as a clinically relevant prognosticator for infertility treatment success. (Fertil Steril (R) 2011;96:663-8. (C)2011 by American Society for Reproductive Medicine.)

Ischemic cecal perforation secondary to ESWL of junctional stone in ureterosigmoidostomy (Mainz Pouch II).

We report an unusual case of ischemic cecal perforation after extracorporeal shock wave lithotripsy for an impacted stone at the distal end of the right ureter of a ureterosigmoidostomy in a 78-year-old female patient.

A polymorphism in the leptin gene promoter is associated with anemia in patients with HIV disease.

To study factors associated with anemia and its effect on survival in HIV-infected persons treated with modern combined antiretroviral therapy (cART), we characterized the prevalence of anemia in the Veterans Aging Cohort Study (VACS) and used a candidate gene approach to identify proinflammatory gene single nucleotide polymorphisms (SNPs) associated with anemia in HIV disease. The study comprised 1597 HIV(+) and 865 HIV(-) VACS subjects with DNA, blood, and annotated clinical data available for analysis. Anemia was defined according to World Health Organization criteria (hemoglobin < 13 g/dL and < 12 g/dL in men and women, respectively). The prevalence of anemia in HIV(+) and HIV(-) subjects was 23.1% and 12.9%, respectively. Independent of HIV status, anemia was present in 23.4% and 8% in blacks and whites, respectively. Analysis of our candidate genes revealed that the leptin -2548 G/A SNP was associated with anemia in HIV(+), but not HIV(-), patients, with the AA and AG genotypes significantly predicting anemia (P < .003 and P < .039, respectively, logistic regression). This association was replicated in an independent cohort of HIV(+) women. Our study provides novel insight into the association between genetic variability in the leptin gene and anemia in HIV(+) individuals.

Síndrome de Burnout entre enfermeiros de um hospital geral da cidade do Recife

Este estudo descritivo, transversal, censitário, identificou Burnout e alguns fatores associados entre enfermeiros da assistência pediátrica e tocoginecológica de hospital geral do nível terciário de atenção do Recife (PE). Participaram 63 profissionais (98,4% do total) que responderam a um questionário auto-aplicável (aspectos sócio-demográficos, condições laborais e Maslach Burnout Inventory). Na análise utilizou-se qui-quadrado, com nível de confiança de 95%. Predominou o gênero feminino (92,1%), com até cinco anos de profissão (68,2%), sendo 52,5% da área pediátrica. Constataram-se altos níveis de exaustão emocional (49,2%) e despersonalização (27,0%) e baixo nível de realização profissional (4,8%), estando 4,7% com Burnout. Mostraram associação: alto nível de exaustão emocional e realizar frequentemente/sempre tarefas com muita rapidez (p=0,039) e receber salário incompatível com o esforço empregado (p=0,016); altos níveis de despersonalização e ter até cinco anos de profissão (p=0,010) e efetuar frequentemente/sempre tarefas com muita rapidez (p=0,009). Para 19,0% pelo menos duas das três dimensões apontavam alta propensão à síndrome.

The influence of Bone Morphogenic Protein-2 on the consolidation phase in a distraction osteogenesis model.

We asked whether locally applied recombinant-Bone Morphogenic Protein-2 (rh-BMP-2) with an absorbable Type I collagen sponge (ACS) carrier could enhance the consolidation phase in a callotasis model. We performed unilateral transverse osteotomy of the tibia in 21 immature male rabbits. After a latency period of 7 days, a 3-weeks distraction was begun at a rate of 0.5mm/12h. At the end of the distraction period (Day 28) animals were randomly divided into three groups and underwent a second surgical procedure: 6 rabbits in Group I (Control group; the callus was exposed and nothing was added), 6 rabbits in Group II (ACS group; receiving the absorbable collagen sponge soaked with saline) and 9 rabbits in Group III (rh-BMP-2/ACS group; receiving the ACS soaked with 100μg/kg of rh-BMP-2, Inductos(®), Medtronic). Starting at Day 28 we assessed quantitative and qualitative radiographic parameters as well as densitometric parameters every two weeks (Days 28, 42, 56, 70 and 84). Animals were sacrificed after 8 weeks of consolidation (Day 84). Qualitative radiographic evaluation revealed hypertrophic calluses in the Group III animals. The rh-BMP-2/ACS also influenced the development of the cortex of the calluses as shown by the modified radiographic patterns in Group III when compared to Groups I and II. Densitometric analysis revealed the bone mineral content (BMC) was significantly higher in the rh-BMP-2/ACS treated animals (Group III).

Multilayered genetic and omics dissection of mitochondrial activity in a mouse reference population

The manner by which genotype and environment affect complex phenotypes is one of the fundamental questions in biology. In this study, we quantified the transcriptome--a subset of the metabolome--and, using targeted proteomics, quantified a subset of the liver proteome from 40 strains of the BXD mouse genetic reference population on two diverse diets. We discovered dozens of transcript, protein, and metabolite QTLs, several of which linked to metabolic phenotypes. Most prominently, Dhtkd1 was identified as a primary regulator of 2-aminoadipate, explaining variance in fasted glucose and diabetes status in both mice and humans. These integrated molecular profiles also allowed further characterization of complex pathways, particularly the mitochondrial unfolded protein response (UPR(mt)). UPR(mt) shows strikingly variant responses at the transcript and protein level that are remarkably conserved among C. elegans, mice, and humans. Overall, these examples demonstrate the value of an integrated multilayered omics approach to characterize complex metabolic phenotypes.

Parent perceived quality of life is age-dependent in children with food allergy.

Food allergy in children significantly affects their quality of life. Its impact can be analyzed by quality of life questionnaires. The aim of our study was to validate the French version of disease-specific questionnaires and to evaluate the quality of life in children with IgE-mediated food allergy. Two validated food allergy-specific questionnaires for quality of life, the parent's and children's forms (FAQLQ-PF and FAQLQ-CF), were translated from English to French and submitted to children with food allergy and their parents. Questionnaires were analyzed in terms of emotional impact, food anxiety, and social and food limitations. NCT 01480427. Sixty-two parents of children aged 0-12 yrs answered the FAQLQ-PF, and 32 children aged 8-12 yrs the FAQLQ-CF. Construct validity of both questionnaires was assessed by correlation between the FAQLQs and FAIM (r = 0.85 and 0.84, respectively). Both FAQLQs had good internal consistency (Cronbach's α = 0.748 and 0.67, respectively). Young children (0-3 yrs old) showed better quality of life scores than older children (FAQLQ-PF global score: p = 0.02). Worse scores were also shown among children with previous severe systemic reactions (FAQLQ-PF global score: p = 0.039), the ones with an allergic mother (FAQLQ-PF global score: p = 0.002), or allergic siblings (FAQLQ-PF emotional impact score: p = 0.034), the ones with multiple food allergy (more than 1 food) (FAQLQ-PF anxiety score: p = 0.04) and among the girls (FAQLQ-CF global score: p = 0.031). Older children, the ones with severe systemic reactions, or with mothers or siblings also affected by allergies, as well as girls, and children with multiple food allergies show worse quality of life scores.

Caracterización ambiental y potencial pesquero 2004-2011: Bahía de Samanco, Chimbote, Perú.

El desarrollo de la maricultura en la Bahía de Samanco ha ocasionado descarga de restos orgánicos al ecosistema marino; se efectuaron monitoreos en esta Bahía en época de veda y de actividad pesquera industrial del 2005 al 2010; temperatura, salinidad y nutrientes se mantuvieron normales, pudiéndose afirmar que la capacidad de resiliencia del ecosistema no está afectada. Los parámetros fisicoquímicos, aceites y grasa presentaron altas concentraciones del 2007 al 2010, en relación al valor límite de los Estándares de Calidad Ambiental. La pesca artesanal generó 21.416 t de recursos hidrobiológicos, conformado por moluscos 57,78%, peces óseos 38,68%, equinodermos 3,20% y crustáceos, celentéreos y algas. La diversidad correspondió a 114 especies, mayor relevancia íctica dada por pejerrey, lisa, machete, lorna y coco; los invertebrados fueron concha de abanico, navajuela, calamar, marucha, caracol y ancoco. La captura de la pesca industrial de madera y artesanal anchovetera para consumo fue de 7.039 t, se identificaron 15 especies: peces (13) e invertebrados (2), la anchoveta aportó 98,71%. Se registraron 11 especies de fitoplancton potencialmente tóxicos, diatomeas: Pseudonitzschia cf delicatissima y P. pungens; dinoflagelados: Alexandrium sp., Dinophysis acuminata, D. caudata, D. rotundata, D. tripos, Gymnodinium sp., Prorocentrum mínimum, Protoperidinium crassipes y P. depressum.

Efeito de sistemas de preparo e de cultura na dinâmica da matéria orgânica e na mitigação das emissões de CO2

Neste estudo, ajustou-se um modelo exponencial de primeira ordem aos dados de carbono orgânico (CO) e N total (NT) do solo, do 5º e 9º ano de um experimento instalado em um Podzólico Vermelho-Escuro, em Eldorado do Sul (RS). Determinaram-se os parâmetros da dinâmica da matéria orgânica e simularam-se os estoques de CO e NT do solo em três sistemas de preparo (convencional-PC, reduzido-PR e plantio direto-PD) e três sistemas de cultura (aveia/milho-A/M, aveia + ervilhaca/milho-A + E/M e aveia + ervilhaca/milho + caupi A + E/M + C). A taxa de decomposição da matéria orgânica do solo diminuiu de 0,054 ano-1, no sistema com lavração e gradagem (PC), para 0,039 ano-1, no solo escarificado (PR), e 0,029 ano-1, no solo não revolvido (PD). Estimou-se que os estoques de CO e NT do solo em 1990 (30,78 Mg ha-1 e 2.200 kg ha-1) diminuirão para 16,11 Mg ha-1 e 1.396 kg ha-1 na combinação PC - A/M. Por sua vez, no sistema PD - A + E/M + C, os estoques de CO e NT do solo em 1990 (32,52 Mg ha-1 e 2.690 Mg ha-1) tenderão a aumentar a estoques estáveis de 54,83 Mg ha-1 e 7.966 kg ha-1, respectivamente. Os sistemas de manejo sem revolvimento do solo e alto aporte de resíduos apresentaram efeito positivo na mitigação das emissões de CO2. Enquanto o sistema PC-A/M apresentou um efluxo líquido (emitido pelo solo > fixado pelas culturas por fotossíntese) de 2,08 Mg CO2 ha-1 para atmosfera no ano de 1994, o sistema PD - A + E/M + C mostrou um influxo líquido (emitido pelo solo < fixado pelas culturas) de CO2 de 1,79 Mg ha-1 ano-1. Atingidos os estoques estáveis de CO no solo estimados pelo modelo, o solo no sistema PD - A + E/M + C terá seqüestrado aproximadamente 142 Mg CO2 ha-1, em comparação ao sistema PC-A/M.

Fator erosividade da chuva de Piraju (SP): distribuição, probabilidade de ocorrência, período de retorno e correlação com o coeficiente de chuva

Foram selecionadas 874 chuvas individuais erosivas, de uma série contínua de 23 anos de registros de dados pluviográficos. As chuvas selecionadas foram cotadas, digitalizadas e, posteriormente, analisadas. O índice de erosividade, EI30, médio anual calculado foi de 7.074 MJ mm ha-1 h-1 ano-1. Espera-se que este índice ocorra no local pelo menos uma vez a cada 2,33 anos, com uma probabilidade de 42,92%. Os valores dos índices anuais de erosividade de Piraju, esperados nos períodos de retorno de 2, 5, 10, 20, 50 e 100 anos, foram, respectivamente, de 6.696, 8.730, 10.076, 11.367, 13.039 e 14.292 MJ mm ha-1 h-1 ano-1. Foi observada uma distribuição de 78,5% do total da erosividade anual durante o semestre de outubro a março, indicando que, nesse período, era esperada a maior parte das perdas de solo por erosão. Observou-se elevada correlação entre o índice de erosividade, EI30, médio mensal e o coeficiente de chuva. Portanto, a equação de regressão determinada permite que seja estimado, com boa margem de segurança, o EI30 para outros locais que não possuam dados pluviográficos, mas que disponham de dados pluviométricos e condições climáticas semelhantes às de Piraju (SP).