Towards a xeno-free and fully chemically defined cryopreservation medium for maintaining viability, recovery, and antigen-specific functionality of PBMC during long-term storage

Analysis of cryopreserved peripheral mononuclear cells (PBMC) is important for evaluating new vaccines in immune based therapies and in pathogenesis studies. To ensure comparable assay results from different laboratories and points of time, collaborative research in multicenter trials needs reliable and reproducible cryopreservation protocols that maintain cell viability and functionality. Current cryomedia consist largely of fetal bovine serum (FBS), a natural mix of growth factors, cytokines, and undefined compounds. Standardized procedures are not possible, as FBS can affect the antigen-specific T-cell response, the most important parameter in functionality assays. Also, worldwide sample exchange is complicated by the strict import restrictions on FBS, because of transfection risk. After establishing a serum-free cryopreservation protocol that maintains cell viability, recovery and antigen-specific T-cell response of PBMC comparably to FBS-based cryomedia (Germann et al., 2011), the aim of this study was the complete avoidance of animal proteins and products in combination with efficient cryopreservation. As long-term stability of the cryopreservation process is crucial for retrospective evaluation of samples at different points of time, PBMC were analyzed after storage for maximal four weeks and again after approximately six months. The cryopreservation efficiency of the protein-free and fully chemically defined cryomedium was comparable to FBS-medium after storage for few weeks and several months. Directly after thawing, this medium yielded viabilities over 97% and recovery values over 84%. Also, the specific T-cell functionality was preserved. Additionally, short-term and six month cryopreservation gave comparable results. The fully chemically defined medium presented here will increase standardization and reproducibility of analysis in multicenter-studies or in retrospective evaluation.     

Poorly differentiated oncocytic thyroid carcinoma--diagnostic implications and outcome

Dettmer M, Schmitt A, Steinert H, Moch H, Komminoth P & Perren A
(2012) Histopathology  60, 1045–1051 Poorly differentiated oncocytic thyroid carcinoma – diagnostic implications and outcome Aims: Poorly differentiated thyroid carcinomas (PDTC) are an ongoing diagnostic challenge. Although the Turin consensus criteria for PDTC excluded consideration of oncocytic tumours, the World Health Organization (WHO) classification does recognise an oncocytic variant of conventional PDTC. The aims of this study were to establish whether the Turin criteria can be applied to oncocytic PDTC, and to determine if there are prognostic differences between conventional and oncocytic PDTC. Methods and results: We applied the Turin criteria to 129 thyroid carcinomas. We identified 18 oncocytic PDTC and 16 conventional PDTC. Kaplan–Meier analysis revealed a significantly worse outcome for oncocytic PDTC with regard to overall and tumour‐specific survival but no difference for relapse‐free survival, all of which were confirmed by multivariate analysis. There was no association of survival with gender, age or tumour stage. Conclusions: The Turin criteria can be applied to oncocytic PDTC and patients with this variant have a decreased survival using conventional radioiodine treatment compared to conventional PDTC and might therefore be candidates for novel treatment modalities.     

Control strategies using diclazuril against coccidiosis in goat kids

Coccidiosis is probably the main parasitic disease affecting goat kids around the weaning period, leading to high economic losses in goat production due to deaths and delayed growth rates of infected animals. A total of 101 kids of 2-4 weeks of age, naturally infected with Eimeria spp., were divided into five groups and studies were conducted to analyse the effects of metaphylactic administration of diclazuril (Vecoxan?) on parasitological and productive parameters. Two different doses of diclazuril (1 and 2 mg/kg BW, p.o.) were given either at 3 weeks (single treatment) or at 3 and 5 weeks of life (double treatment). The faecal oocyst shedding and the body weights of the animals were monitored at 2-weeks intervals for 6 consecutive weeks. Treatments of goat kids with diclazuril were effective against the three most predominant Eimeria species recorded in this study (E. arloingi, E. ninakohlyakimovae and E. christenseni) and also against other minor species found in faecal examinations, including E. alijevi, E. caprina, E. jolchijevi, E. caprovina, E. hirci and E. aspheronica). In consequence, OPG values lower than 1?×?10(3) were detected in 90 to 100% of the animals up to 15-20 days post-treatment depending on the treatment regimen. Even a single dose of 1 mg/kg BW p.o. resulted in an increase of growth rates in treated animals and therefore should be considered as a control strategy in farms precluding coccidian infections, whilst double and multiple dose treatments could be the recommendation for environments heavily contaminated with Eimeria oocysts. In relation to the OPG reduction and increased growth rates, the severity of the clinical signs (i.e., diarrhoea) was ameliorated in treated animals during the course of infection compared to that of non-treated or control kids. The precise timing of treatment appears crucial in order to prevent severe clinical coccidiosis and thereby enabling the adequate development of protective immune response against Eimeria challenge infections.     

The contribution of risk factors to the higher incidence of invasive and in situ breast cancers in women with higher levels of education in the European prospective investigation into cancer and nutrition

The authors investigated the role of known risk factors in educational differences in breast cancer incidence. Analyses were based on the European Prospective Investigation Into Cancer and Nutrition and included 242,095 women, 433 cases of in situ breast cancer, and 4,469 cases of invasive breast cancer. Reproductive history (age at first full-term pregnancy and parity), exposure to endogenous and exogenous hormones, height, and health behaviors were accounted for in the analyses. Relative indices of inequality (RII) for education were estimated using Cox regression models. A higher risk of invasive breast cancer was found among women with higher levels of education (RII = 1.22, 95% confidence interval (CI): 1.09, 1.37). This association was not observed among nulliparous women (RII = 1.13, 95% CI: 0.84, 1.52). Inequalities in breast cancer incidence decreased substantially after adjusting for reproductive history (RII = 1.11, 95% CI: 0.98, 1.25), with most of the association being explained by age at first full-term pregnancy. Each other risk factor explained a small additional part of the inequalities in breast cancer incidence. Height accounted for most of the remaining differences in incidence. After adjusting for all known risk factors, the authors found no association between education level and risk of invasive breast cancer. Inequalities in incidence were more pronounced for in situ breast cancer, and those inequalities remained after adjustment for all known risk factors (RII = 1.61, 95% CI: 1.07, 2.41), especially among nulliparous women.     

Fibre intake among the Belgian population by sex-age and sex-education groups and its association with BMI and waist circumference

The objectives of the present study were to assess total dietary fibre intake and the main contributors to fibre intake in the Belgian population by sex-age and sex-education groups and to investigate its relationship with BMI and waist circumference (WC). The participants of the Belgian food consumption survey (2004) were randomly selected. Information about food intake was collected using two repeated, non-consecutive 24 h recall interviews. A total of 3083 individuals ( ≥ 15 years; 1546 men and 1537 women) completed both interviews. The main contributors to total fibre intake (17·8 g/d) were cereals and cereal products (34 %; 5·9 g/d), potatoes and other tubers (18·6 %; 3·3 g/d), fruits (14·7 %; 2·8 g/d) and vegetables (14·4 %; 2·6 g/d). Legume fibre intake was extremely low (0·672 %; 0·139 g/d). In all sex-age and sex-education groups, total fibre intake was below the recommendations of the Belgian Superior Health Council. Men (21 g/d) consumed significantly more fibre than women (17·3 g/d) (P < 0·001). Lower educated men and higher educated women reported the highest fibre intake. A significant inverse association was found between total fibre intake and WC (β = - 0·118, P < 0·001). Fruit-derived fibre was positively associated with WC (β = 0·731, P = 0·001). In summary, total fibre intake was inversely associated with WC, whereas fruit-derived fibre intake was positively associated with WC in the Belgian population.     

Plant and animal protein intake and its association with overweight and obesity among the Belgian population

The objective of the present study was to assess animal and plant protein intakes in the Belgian population and to examine their relationship with overweight and obesity (OB). The subjects participated in the Belgian National Food Consumption Survey conducted in 2004. Food consumption was assessed by using two non-consecutive 24?h dietary recalls. About 3083 participants (?≥?15 years of age; 1546 males, 1537 females) provided completed dietary information. Animal protein intake (47?g/d) contributed more to total protein intakes of 72?g/d than plant protein intake, which accounted for 25?g/d. Meat and meat products were the main contributors to total animal protein intakes (53?%), whereas cereals and cereal products contributed most to plant protein intake (54?%). Males had higher animal and plant protein intakes than females (P?相似文献   

Androgen and androgen metabolite levels in serum and urine of East African chimpanzees (Pan troglodytes schweinfurthii): comparison of EIA and LC-MS analyses

The primary male androgen testosterone (T) is often used as an endocrinological marker to investigate androgen-behaviour interactions in males. In chimpanzees and bonobos, studies investigating the relationship between T levels and dominance rank or aggressive behaviour have revealed contradictory results. The immunoassays used in these studies were originally developed for the measurement of steroids in serum. Their application to non-invasively collected samples, however, can lead to methodological problems due to cross-reacting metabolites, which might occur in urine or faeces but not in blood. The overall aim of this study, therefore, is to clarify whether a T enzyme immunoassay (EIA) is an applicable method to monitor testicular function in adult male chimpanzees. To estimate the impact of cross-reacting androgens on the used T EIA, we compared the results of an EIA measurement with a set of androgen metabolite levels measured by LC–MS. In urine from male chimpanzees, cross-reactivities appear to exist mainly with T and its exclusive metabolites, 5α-dihydrotestosterone (5α-DHT) and 5α-androstanediol (androstanediol). Both urinary and serum T levels of male chimpanzees were significantly higher than female T levels when measured with the T EIA, indicating a reliable measurement of testicular androgens and their exclusive metabolites with the used EIA. In urine from female chimpanzees, the comparison between LC–MS and T EIA results indicated a higher impact of cross-reactions with adrenal androgen metabolites. Therefore, the investigation of urinary T levels in female chimpanzees with a T EIA seems to be problematic. Overall our results show that a T EIA can be a reliable method to monitor testicular function in male chimpanzee urine and that LC–MS is a valuable tool for the validation of immunoassays.