Association between Psychosocial Stress and Fecal Microbiota in Pregnant Women.

Maternal prenatal psychosocial stress is associated with altered child emotional and behavioral development. One potential underlying mechanism is that prenatal psychosocial stress affects child outcomes via the mother's, and in turn the child's, intestinal microbiota. This study investigates the first step of this mechanism: the relation between psychosocial stress and fecal microbiota in pregnant mothers. Mothers (N = 70) provided a late pregnancy stool sample and filled in questionnaires on general and pregnancy-specific stress and anxiety. Bacterial DNA was extracted and analysed by Illumina HiSeq sequencing of PCR-amplified 16 S ribosomal RNA gene fragments. Associations between maternal general anxiety and microbial composition were found. No associations between the other measured psychosocial stress variables and the relative abundance of microbial groups were detected. This study shows associations between maternal pregnancy general anxiety and microbial composition, providing first evidence of a mechanism through which psychological symptoms in pregnancy may affect the offspring.

In vitro cultured fetal fibroblasts have myofibroblast-associated characteristics and produce a fibrotic-like environment upon stimulation with TGF-ß1: Is there a thin line between fetal scarless healing and fibrosis?

Transforming growth factor-ß (TGF-ß) is a cytokine occurring in three isoforms with an important function in development and wound healing. In wound healing, prolonged TGF-ß signaling results in myofibroblast differentiation and fibrosis. In contrast, the developing second-trimester fetal skin contains high levels of all three TGF-ß isoforms but still has the intrinsic capacity to heal without scarring. Insight into TGF-ß signal transduction during fetal wound healing might lead to methods to control the signaling pathway during adult wound healing. In this study, we imitated wound healing in vitro by stimulating fibroblasts with TGF-ß1 and examining myofibroblast differentiation. The aim was to gain insight into TGF-ß signaling in human fibroblasts from fetal and adult dermis. First, TGF-ß1 stimulation resulted in similar or even more severe upregulation of myofibroblast-associated genes in fetal fibroblasts compared to adult fibroblasts. Second, fetal fibroblasts also had higher protein levels of myofibroblast-marker α-smooth muscle actin (α-SMA). Third, stimulated fetal fibroblasts in collagen matrices had higher protein levels of α-SMA, produced more of the fibrotic protein fibronectin splice-variant extra domain A (FnEDA), and showed enhanced contraction. Finally, fetal fibroblasts also produced significant higher levels of TGF-ß1. Altogether, these data show that in vitro cultured fetal fibroblasts have myofibroblast-associated characteristics and do produce a fibrotic environment. As healthy fetal skin has high levels of TGF-ß1, FnEDA, and collagen-III as well, these findings correlate with the in vivo situation. Therefore, our study demonstrates that there are similarities between fetal skin development and fibrosis and shows the necessity to discriminate between these processes.

Transforming growth factor-ß (TGF-ß) signaling in healthy human fetal skin: a descriptive study.

BACKGROUND: TGF-ß plays an important role in growth and development but is also involved in scarring and fibrosis. Differences for this growth factor are known between scarless fetal wound healing and adult wound healing. Nonetheless, most of the data in this area are from animal studies or in vitro studies and, thus, information about the human situation is incomplete and scarce. OBJECTIVE: The aim of this study was to compare the canonical TGF-ß signaling in unwounded human fetal and adult skin. METHODS: Q-PCR, immunohistochemistry, Western Blot and Luminex assays were used to determine gene expression, protein levels and protein localization of components of this pathway in healthy skin. RESULTS: All components of the canonical TGF-ß pathway were present in unwounded fetal skin. Compared to adult skin, fetal skin had differential concentrations of the TGF-ß isoforms, had high levels of phosphorylated receptor-Smads, especially in the epidermis, and had low expression of several fibrosis-associated target genes. Further, the results indicated that the processes of receptor endocytosis might also differ between fetal and adult skin. CONCLUSION: This descriptive study showed that there are differences in gene expression, protein concentrations and protein localization for most components of the canonical TGF-ß pathway between fetal and adult skin. The findings of this study can be a starting point for further research into the role of TGF-ß signaling in scarless healing.

Symptoms from treatment with sunitinib or sorafenib: a multicenter explorative cohort study to explore the influence of patient-reported outcomes on therapy decisions.

PURPOSE: Optimal long-lasting treatment with sunitinib and sorafenib is limited by dose modifications (DMs) due to adverse events (AEs). These AEs may be underrecognized and their influence on health-related quality of life (HRQL) underestimated. Improved insight into the relationship between AEs and therapy decisions is needed. To improve decision making around managing symptoms and reduce DMs, this study was set up to explore the influence of patient-reported symptoms on therapy decisions. METHODS: In this multicenter cohort study, patient characteristics, reasons for and different forms of used dose modifications, and AEs were prospectively obtained from cancer patients on sunitinib/sorafenib treatment. Used instruments to get insight into AEs were the patient-scored Utrecht Symptom Diary (USD) and the professional-scored Common Terminology Criteria for AEs version 3.0. RESULTS: Median total treatment duration in 42 patients was 16 weeks. Median time till dose modification was 10 weeks. DMs occurred mostly due to multiple mild AEs. By using the USD, a higher prevalence of most AEs was found compared to the literature. Sixty percent of the patients experienced a decreased HRQL due to multiple AEs. CONCLUSIONS: Because severe AEs due to sunitinib/sorafenib treatment seldom occur, it is more important to focus on treating and preventing multiple mild AEs with higher impact on HRQL, when trying to avoid dose modifications. Using patient self-reported measurement methods helps to early recognize symptoms and to differentiate among symptom intensities. This systematic approach might help to achieve the optimal dosing, which might improve PFS and OS.

Targeting the endothelin axis with atrasentan, in combination with IFN-alpha, in metastatic renal cell carcinoma.

BACKGROUND: The endothelin system is involved in tumour growth. Atrasentan, a selective endothelin-A-receptor antagonist, blocks endothelin signalling. This phase I trial studied combining treatment of interferon-alpha (IFN-α) with atrasentan in renal cell carcinoma (RCC). PATIENTS AND METHODS: This study evaluated the safety and tolerance of IFN-α (9MU subcutaneously (s.c.) three times a week) in combination with atrasentan (2.5, 5 and 10 mg orally once daily) in untreated metastatic RCC. Cohort 10 mg was extended to obtain insights in efficacy and pharmacodynamics. RESULTS: Observed toxicities mainly consisted of known IFN-like toxicities (anorexia, chills, fever, fatigue and nausea), and of nasal congestion (associated to atrasentan). None of these toxicities were considered dose limiting. Cohort 10 mg was extended up to 32 patients; in a subset of patients treated according to the protocol (n=27), median overall survival (OS) was 17.3 months. One patient (3.1%) showed a partial response lasting 14.3 months. In an exploratory analysis, we observed that in the subset of patients with declining vascular endothelial growth factor (VEGF) levels (in combination with rising Endothelin-1 levels), median OS was 22.2 months compared with 2.2 months in patients with increasing VEGF levels. CONCLUSION: Combination treatment of IFN-α 9MU-α s.c. three times a week and atrasentan 10 mg once daily is tolerated. Clinical activity, especially OS, and biomarkers in our view warrant further studies targeting the endothelin axis.

Simple and selective method for the determination of various tyrosine kinase inhibitors used in the clinical setting by liquid chromatography tandem mass spectrometry.

A fast, sensitive, universal and accurate method for the determination of four different tyrosine kinase inhibitors from biological material was developed using LC-MS/MS techniques. Utilizing a simple protein precipitation with acetonitrile a 20 microl sample volume of biological matrixes can be extracted at 4 degrees C with minimal effort. After centrifugation the sample extract is introduced directly onto the LC-MS/MS system without further clean-up and assayed across a linear range of 1-4000 ng/ml. Chromatography was performed using a Dionex Ultimate 3000 with a Phenomenex prodigy ODS3 (2.0 mm x 100 mm, 3 microm) column and eluted at 200 microl/min with a tertiary mobile phase consisting of 20mM ammonium acetate:acetonitrile:methanol (2.5:6.7:8.3%). Injection volume varied from 0.1 microl to 1 microl depending on the concentration of the drug observed. Samples were observed to be stable for a maximum of 48 h after extraction when kept at 4 degrees C. Detection was performed using a turbo-spray ionization source and mass spectrometric positive multi-reaction-monitoring-mode (+MRM) for Gefitinib (447.1 m/z; 127.9 m/z), Erlotinib (393.9 m/z; 278.2 m/z), Sunitinib (399.1 m/z; 283.1 m/z) and Sorafenib (465.0 m/z; 251.9 m/z) at an ion voltage of +3500 V. The accuracy, precision and limit-of-quantification (LOQ) from cell culture medium were as follows: Gefitinib: 100.2+/-3.8%, 11.2 nM; Erlotinib: 101.6+/-3.7%, 12.7 nM; Sunitinib: 100.8+/-4.3%, 12.6 nM; Sorafenib: 93.9+/-3.0%, 10.8 nM, respectively. This was reproducible for plasma, whole blood, and serum. The method was observed to be linear between the LOQ and 4000 ng/ml for each analyte. Effectiveness of the method is illustrated with the analysis of samples from a cellular accumulation investigation and from determination of steady state concentrations in clinically treated patients.

[Three patients with gynaecomastia]. / Drie patiënten met gynaecomastie.

In three patients gynaecomastia was diagnosed: a 22-year-old man with concomitant thyrotoxicosis due to an extensively metastasized extragonadal choriocarcinoma, a 53-year-old man with hypogonadism due to Klinefelter's syndrome that was biochemically obscured due to medications leading to elevated prolactin levels, and a 62-year-old man with acromegaly and secondary hypogonadism due to a mixed prolactin and growth hormone secreting pituitary adenoma. Gynaecomastia calls for thorough evaluation.

Antibody responses and body weights of chicken lines selected for high and low humoral responsiveness to sheep red blood cells. 2. Effects of separate application of Freund's Complete and Incomplete Adjuvant and antigen.

Antibody (Ab) responses to SRBC, BSA, Mycobacterium butyricum, and Escherichia coli lipopolysaccharide (LPS) were measured in two chicken lines divergently selected for high and low Ab responses to SRBC, and in a randombred control line. Levels of Ab binding SRBC, BSA, and Mycobacterium protein, but not LPS were higher in the high Ab producing (H) line than in the control (C) and low Ab producing (L) lines (P < 0.05), and at almost every time, the L line showed significantly lower titers than the H and C lines. In the H and C lines, Ab responses to SRBC were enhanced when Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA) were simultaneously administered on a separate location than SRBC. In the L line, Ab titers to SRBC and BSA were enhanced when antigen was administered emulsified in CFA. At all times until 28 d after sensitization the C and L line birds were significantly heavier than birds of the H line. Body weight, body growth, and percentage body growth were impaired in birds that received antigen emulsified in CFA, which suggested a negative relationship between BW gain and immune responses to Mycobacteria protein. Prolonged divergent selection for Ab responses to SRBC resulted into two lines that not only differ in Ab responses to T cell-dependent antigens but also in BW. In contrast to previous findings with the current lines, line differences with respect to Ab responses were not abolished by CFA treatment.

Antibody responses and body weights of chicken lines selected for high and low humoral responsiveness to sheep red blood cells. 1. Effect of Escherichia coli lipopolysaccharide.

Antibody (Ab) responses to i.m. administered SRBC and BSA, and i.p. administered Escherichia coli lipopolysaccharide (LPS), and BW at various times after treatment, were measured in chicken lines divergently selected for high (H) and low (L) Ab responses to SRBC, and in a randombred control line (C). The Ab responses to SRBC and BSA, but not LPS, were significantly affected by line by treatment interactions. Levels of antibodies to SRBC and BSA were higher in the H line than in either the C or L line (P < 0.05). Administration of LPS did not affect Ab responses to SRBC, but Ab responses to BSA were decreased in birds that received BSA and LPS simultaneously. Body weights of C and L lines were significantly higher than BW of H line birds at all times. Lipopolysaccharide injection induced an acute, but transient reduction of BW gain, which was not affected by line. Antibody responses to SRBC and BSA were negatively correlated with BW. During the experimental period, however, percentage BW gain and humoral responsiveness were positively correlated. A higher percentage BW gain growth was seen in H line birds at the end of the experimental period. The present results confirm the hypothesized acute cachectin nature of LPS, but the relationship between live BW (gain) and immune responsiveness in chickens remains to be further clarified.