Kinetics of human myeloid-derived suppressor cells after blood draw.

BACKGROUND: Human myeloid-derived suppressor cells (MDSC) have been described as a group of immature myeloid cells which exert immunosuppressive action by inhibiting function of T lymphocytes. While there is a huge scientific interest to study these cells in multiple human diseases, the methodological approach varies substantially between published studies. This is problematic as human MDSC seem to be a sensible cell type concerning not only cryopreservation but also time point after blood draw. To date data on delayed blood processing influencing cell numbers and phenotype is missing. We therefore evaluated the kinetics of granulocytic MDSC (gMDSC) and monocytic MDSC (mMDSC) frequencies after blood draw in order to determine the best time point for analysis of this recently defined cell type. METHODS: In this study, we isolated peripheral blood mononuclear cells (PBMC) of patients with HIV infection or solid tumors directly after blood draw. We then analyzed the frequencies of gMDSC and mMDSC 2, 4 and 6 h after blood draw and after an overnight rest by FACS analysis using the standard phenotypic markers. In addition, part of the cells was frozen directly after PBMC preparation and was measured after thawing. RESULTS: gMDSC levels showed no significant difference using fresh PBMC over time with a limitation for the overnight sample. However they were massively diminished after freezing (p = 0.0001 for all subjects). In contrast, frequencies of fresh mMDSC varied over time with no difference between time point 2 and 4 h but a significantly reduction after 6 h and overnight rest (p = 0.0005 and p = 0.005 respectively). Freezing of PBMC decreased the yield of mMDSC reaching statistical significance (p = 0.04). For both MDSC subgroups, FACS analysis became more difficult over time due to less sharp divisions between populations. CONCLUSIONS: According to our data human MDSC need to be studied on fresh PBMC. gMDSC can be studied with delay, mMDSC however should be studied no later than 4 h after blood draw. These results are crucial as an increasing number of clinical trials aim at analyzing MDSC nowadays and the logistics of blood processing implies delayed sample processing in some cases.

Reduced sleep quality and depression associate with decreased quality of life in patients with pituitary adenomas.

OBJECTIVES: Several studies reported decreased quality of life (QoL) and sleep as well as increased rates of depression for patients with pituitary adenomas. Our aim was to explore to what extent differences in depression and sleep quality contribute to differences in QoL between patients with pituitary adenomas and controls. DESIGN: A cross-sectional case-control study. SETTING: Endocrine Outpatient Unit of the Max Planck Institute of Psychiatry, Munich, Department of Internal Medicine, Ludwig-Maximilians-University, Munich, and the Institute of Clinical Psychology and Psychotherapy, Technical University, Dresden. PARTICIPANTS: Patients with pituitary adenomas (n=247) and controls (from the DETECT cohort, a large epidemiological study in primary care patients) matched individually by age and gender (n=757). MEASUREMENTS: Sleep quality was assessed with the Pittsburgh Sleep Quality Index (PSQI) and QoL was measured by the generic EQ-5D and calculated by the time trade-off- and VAS-method. Depression was categorized as 'no depression', 'subclinical depression', and 'clinical depression' according to the Beck Depressions Inventory for patients and the Depression Screening Questionnaire for control subjects. STATISTICAL ANALYSES: General linear and generalized, logistic mixed models as well as proportional odds mixed models were calculated for analyzing differences in baseline characteristics and in different subgroups. RESULTS: Patients with pituitary adenomas showed decreased QoL (VAS index: 0.73±0.19) and sleep (PSQI score: 6.75±4.17) as well as increased rates of depression (subclinical or clinical depression: 41.4%) compared with their matched control subjects (VAS index: 0.79±0.18, PSQI score: 5.66±4.31, subclinical or clinical depression: 25.9%). We have shown that a substantial proportion of the reduced QoL (48% respectively 65%) was due to the incidence of depression and reduced sleep quality. CONCLUSIONS: These findings emphasize the importance of diagnosing depressive symptoms and sleep disturbances in patients with pituitary disease, with the ultimate goal to improve QoL in patients with pituitary adenomas.

Insights into the coexistence of two mutations in the same LHCGR gene locus causing severe Leydig cell hypoplasia.

BACKGROUND: Leydig cell hypoplasia is a rare autosomal recessive condition that interferes with the normal development of male external genitalia in 46,XY individuals. It is mediated by mutations in the lutropin/choriogonadotropin receptor gene, resulting in the impairment of either the binding of hormone or signal transduction. OBJECTIVE/DESIGN: We report a 32-year-old female patient with severe Leydig cell hypoplasia due to a novel homozygote nonsense mutation in exon 10 (c.907C>T, p.Gln303Ter) of the lutropin/choriogonadotropin receptor gene. Interestingly, a second mutation was found (c.935A>G, p.Asn312Ser) downstream of the disruption of the gene sequence. CONCLUSIONS: This case report demonstrates the coexistence of a novel homozygote nonsense mutation with a second mutation in the same hormone binding domain, expanding the genotypic spectrum of lutropin-choriogonadotropic hormone receptor gene mutations. The first diagnosis of this mutation in an adult 46,XY female patient from Morocco underlines the importance of thorough clinical and genetic examination, not only in pre- and post-pubertal children but also in adults originating from conservative socio-cultural backgrounds.

Twenty years of endocrinologic treatment in transsexualism: analyzing the role of chromosomal analysis and hormonal profiling in the diagnostic work-up.

OBJECTIVE: To demonstrate that adequate pubertal history, physical examination, and a basal hormone profile is sufficient to exclude disorders of sexual development (DSD) in adult transsexuals and that chromosomal analysis could be omitted in cases of unremarkable hormonal profile and pubertal history. DESIGN: Retrospective chart analysis. SETTING: Endocrine outpatient clinic of a psychiatric research institute. PATIENT(S): A total of 475 subjects (302 male-to-female transsexuals [MtF], 173 female-to-male transsexuals [FtM]). Data from 323 (192 MtF/131 FtM) were collected for hormonal and pubertal abnormalities. Information regarding chromosomal analysis was available for 270 patients (165 MtF/105 FtM). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pubertal abnormalities, menstrual cycle, and hormonal irregularities in relation to chromosomal analysis conducted by karyotype or hair root analysis. RESULT(S): In the MtF group, 5.2% of the patients reported pubertal irregularities and 5.7% hormonal abnormalities, and in the FtM group 3.8% and 19.1%, respectively. Overall chromosomal abnormality in both groups was 1.5% (2.9% in the FtM and 0.6% in the MtF group). The aneuploidies found included one gonosomal aneuploidy (45,X[10]/47,XXX[6]/46,XX[98]), two Robertsonian translocations (45,XXder(14;22)(q10;q10)), and one Klinefelter syndrome (47,XXY) that had already been diagnosed in puberty. CONCLUSION(S): Our data show a low incidence of chromosomal abnormalities and thus question routine chromosomal analysis at the baseline evaluation of transsexualism, and suggest that it be considered only in cases of abnormal history or hormonal examination.

Inadequate statistical power of published comparative cohort studies on ventilator-associated pneumonia to detect mortality differences.

BACKGROUND: Comparative cohort studies are often conducted to identify novel therapeutic strategies or prognostic factors for ventilator-associated pneumonia (VAP). We aimed to evaluate the power of such studies to provide clinically and statistically significant conclusions with regard to mortality differences. METHODS: We searched in PubMed and Scopus for comparative cohort studies that evaluated mortality in patients with VAP. We calculated the central estimates and corresponding 95% confidence intervals (CIs) for mortality differences between compared patient groups. We also calculated the statistical power of the included studies to detect a difference in mortality that corresponds to a risk ratio of 0.80. RESULTS: We identified 39 (20 prospective) comparative cohort studies on VAP as eligible for inclusion in this analysis. The median absolute risk difference in mortality between compared groups was 10% (interquartile range [IQR], 5%-18%), and the median width of the 95% CI of the absolute risk difference in mortality was 34% (IQR, 28%-42.5%). The median power of the included studies to detect a risk ratio for mortality of 0.80 was 14.7% (IQR, 10.6%-21.8%). CONCLUSIONS: There is considerable uncertainty around the central estimate of comparative cohort studies on VAP with regard to mortality differences. For a wiser use of resources allocated to research, we emphasize the need to conduct cohort studies with larger sample size so that potential differences between the compared groups are more likely to be shown.

Prevalence of hepatitis B, hepatitis C, and HIV infections among patients in a psychiatric hospital in Greece.

OBJECTIVE: Psychiatric patients are considered to be at increased risk of infection with hepatitis B virus (HBV), hepatitis C virus (HCV), or HIV. METHODS: This study retrospectively assessed the seroprevalence of these infections among psychiatric patients who were referred for laboratory testing over a 24-month period in a 415-bed tertiary care psychiatric hospital in Greece. RESULTS: More than two-thirds of the 805 unique tested patients received care in short-term hospitalization units (patients receiving treatment for substance-related disorders were excluded from this analysis). Two percent tested positive for having the hepatitis B surface antigen, 9% for the HCV antibody, and 1% for HIV. Males were more likely than females to test positive for the HCV antibody (p=.04). CONCLUSIONS: Compared with rates in the general population, in this study population the rate of HCV was more than ten times as high, the rate of HIV was three times as high, and the rate of HBV was comparable.

Mapping health literacy research in the European Union: a bibliometric analysis.

BACKGROUND: To examine and compare the research productivity on selected fields related to health literacy of the current members of the European Union, the four candidate countries waiting to join the EU, Norway, Switzerland, and the United States. METHODOLOGY/PRINCIPAL FINDINGS: A bibliometric analysis (1991-2005). Data sources included papers published by authors from each country separately. The 25 European countries produce less than 1/3 health literacy research when compared to the U.S. (13,710 and 49,523 articles were published by authors with main affiliation in the European Union and the four candidate countries, and the U.S., respectively). The Netherlands and Sweden (followed by Germany, Italy, and France) are the European countries with the highest number of research published in fields related to health literacy. After adjustment for population Sweden, Finland, and Norway, were on the top of the relevant list. In addition, Sweden, Finland, and Ireland, were on the top of the list of countries regarding research productivity on the selected fields after adjustment for gross domestic product (GDP). CONCLUSIONS/SIGNIFICANCE: Inequalities in research published on the topic of health literacy exist among Europe, Norway, Switzerland, and the U.S. More research may need to be done in all areas of health literacy in Europe and the potential detrimental effects of this gap should be further investigated.