ENDOCELL-Seud : a Delphi protocol to harmonise methods in endometrial cell culturing

culturing of endometrial cells obtained from the uterine mucosa or ectopic sites is used to study molecular and cellular signalling relevant to physiologic and pathologic reproductive conditions. However, the lack of consensus on standard operating procedures for deriving, characterising and maintaining primary cells in two- or three-dimensional cultures from eutopic or ectopic endometrium may be hindering progress in this area of research. Guidance for unbiased in vitro research methodologies in the field of reproductive science remains essential to increase confidence in the reliability of in vitro models. We present herein the protocol for a Delphi process to develop a consensus on in vitro methodologies using endometrial cells (ENDOCELL-Seud Project). A steering committee composed of leading scientists will select critical methodologies, topics and items that need to be harmonised and that will be included in a survey. An enlarged panel of experts (ENDOCELL-Seud Working Group) will be invited to participate in the survey and provide their ratings to the items to be harmonised. According to Delphi, an iterative investigation method will be adopted. Recommended measures will be finalised by the steering committee. The study received full ethical approval from the Ethical Committee of the Maastricht University (ref. FHML-REC/2021/103). The study findings will be available in both peer-reviewed articles and will also be disseminated to appropriate audiences at relevant conferences

IL-21 induces in vivo immune activation of NK cells and CD8+ T cells in patients with metastatic melanoma and renal cell carcinoma

PURPOSE: Human interleukin-21 (IL-21) is a class I cytokine previously reported in clinical studies on immune responsive cancers. Here we report the effects of systemic IL-21 therapy on the immune system in two phase 1 trials with this novel cytokine. EXPERIMENTAL DESIGN: Recombinant IL-21 was administered by intravenous bolus injection at dose levels from 1 to 100 microg/kg using two planned treatment regimens: thrice weekly for 6 weeks (3/week); or once daily for five consecutive days followed by nine dose-free days (5 + 9). The following biomarkers were studied in peripheral blood mononuclear cells (PBMC) during treatment: phosphorylation of STAT3, alterations in the composition of leukocyte subsets, ex vivo cytotoxicity, expression of effector molecules in enriched CD8(+) T cells and CD56(+) NK cells by quantitative RT-PCR, and gene array profiling of CD8(+) T cells. RESULTS: Effects of IL-21 were observed at all dose levels. In the 5 + 9 regimen IL-21 induced a dose dependent decrease in circulating NK cells and T cells followed by a return to baseline in resting periods. In both CD8(+) T cells and CD56(+) NK cells we found up-regulation of perforin and granzyme B mRNA. In addition, full transcriptome analysis of CD8(+) T cells displayed changes in several transcripts associated with increased cell cycle progression, cellular motility, and immune activation. Finally, cytotoxicity assays showed that IL-21 enhanced the ability of NK cells to kill sensitive targets ex vivo. CONCLUSIONS: IL-21 was biologically active at all dose levels administered with evidence of in vivo NK cell and CD8(+) T cell activation

Anamnestic risk factor questionnaire as reliable diagnostic instrument for osteoporosis (reduced bone morphogenic density)

<p>Abstract</p> <p>Background</p> <p>Osteoporosis is a major health problem worldwide, and is included in the WHO list of the top 10 major diseases. However, it is often undiagnosed until the first fracture occurs, due to inadequate patient education and lack of insurance coverage for screening tests. Anamnestic risk factors like positive family anamnesis or early menopause are assumed to correlate with reduced BMD.</p> <p>Methods</p> <p>In our study of 78 patients with metaphyseal long bone fractures, we searched for a correlation between anamnestic risk factors, bone specific laboratory values, and the bone morphogenic density (BMD). Each indicator was examined as a possible diagnostic instrument for osteoporosis. The secondary aim of this study was to demonstrate the high prevalence of osteoporosis in patients with metaphyseal fractures.</p> <p>Results</p> <p>76.9% of our fracture patients had decreased bone density and 43.6% showed manifest osteoporosis in DXA (densitometry) measurements. Our questionnaire, identifying anamnestic risk factors, correlated highly significantly (p = 0.01) with reduced BMD, whereas seven bone-specific laboratory values (p = 0.046) correlated significantly.</p> <p>Conclusions</p> <p>Anamnestic risk factors correlate with pathological BMD. The medical questionnaire used in this study would therefore function as a cost-effective primary diagnostic instrument for identification of osteoporosis patients.</p

Accelerometer Measured Levels of Moderate-to-Vigorous Intensity Physical Activity and Sedentary Time in Children and Adolescents with Chronic Disease: a Systematic Review and Meta-Analysis

Context: Moderate-to-vigorous physical activity (MVPA) and sedentary time (ST) are important for child and adolescent health. Objective: To examine habitual levels of accelerometer measured MVPA and ST in children and adolescents with chronic disease, and how these levels compare with healthy peers. Methods: Data sources: An extensive search was carried out in Medline, Cochrane library, EMBASE, SPORTDiscus and CINAHL from 2000–2017. Study selection: Studies with accelerometer-measured MVPA and/or ST (at least 3 days and 6 hours/day to provide estimates of habitual levels) in children 0–19 years of age with chronic diseases but without co-morbidities that would present major impediments to physical activity. In all cases patients were studied while well and clinically stable. Results: Out of 1592 records, 25 studies were eligible, in four chronic disease categories: cardiovascular disease (7 studies), respiratory disease (7 studies), diabetes (8 studies), and malignancy (3 studies). Patient MVPA was generally below the recommended 60 min/day and ST generally high regardless of the disease condition. Comparison with healthy controls suggested no marked differences in MVPA between controls and patients with cardiovascular disease (1 study, n = 42) and type 1 diabetes (5 studies, n = 400; SMD -0.70, 95% CI -1.89 to 0.48, p = 0.25). In patients with respiratory disease, MVPA was lower in patients than controls (4 studies, n = 470; SMD -0.39, 95% CI -0.80, 0.02, p = 0.06). Meta-analysis indicated significantly lower MVPA in patients with malignancies than in the controls (2 studies, n = 90; SMD -2.2, 95% CI -4.08 to -0.26, p = 0.03). Time spent sedentary was significantly higher in patients in 4/10 studies compared with healthy control groups, significantly lower in 1 study, while 5 studies showed no significant group difference. Conclusions: MVPA in children/adolescents with chronic disease appear to be well below guideline recommendations, although comparable with activity levels of their healthy peers except for children with malignancies. Tailored and disease appropriate intervention strategies may be needed to increase MVPA and reduce ST in children and adolescents with chronic disease

Development of light-weight steel castings for efficient aircraft engines - Summary report

This report summaries the work carried out in the project, LEAN - Development of light-weight steel castings for efficient aircraft engines. The overall object in the project was to develop thinner cast steel components for aircraft engines. The work was conducted in collaboration between research institutes and industry. Swerea SWECAST, the Swedish foundry institute, which has a long tradition in collaboration with the industry in research projects, was the project leader and leader in one of the work packages. FRI, the Polish Foundry Research Institute, which has advance equipment for material science and material data investigations, was the leader of the second work package. The investment foundry TPC Components AB conducted casting trials and contributed with their wide knowledge of investment casting. Casting trials have been performed in order to investigate the influence of different process parameters governing the fluidity of thin walled investment castings. The alloy used was CbCu7-1, i.e. the cast analogy of the stainless precipitation-hardening steel 17-4PH. Two levels of geometry complexity were used as well as top- and bottom gated casting systems. In the first trial, a blade thicknesses ranging from 0.7-2.0 mm was used. In the second trial, some features were added to the blade as well as a textured surface on one side. It was shown that the top gated casting system showed an overall improved fluidity compared to the bottom gated casting system for the simpler geometry. Blade thickness and pouring temperature were shown to have the greatest impact on fluidity. Adding some geometrical features to the simple geometry drastically decreased the differences between the filling systems. Using a one side textured blade with thickness of 1.3 and 1.5 mm was comparable with 1.5 and 2.0 mm flat castings thus reducing weight of the thinnest sections of a steel casting. Predictions of miss-runs with simulations were shown to be in good agreement with experiments and gave valuable insight to problems in the casting trials. Differences in porosity levels were seen between the top- and bottom gated casting systems for the simpler geometry at high metal temperatures, where the former showed a larger amount of porosity. Besides the work performed on fluidity of the cast analogy of 17-4PH, a number of other alloys not commonly used for castings today were evaluated in terms of their fluidity and were compared to 17-4PH. The casting trials were performed with a bar casting. It was shown that JETHETE 152M had the best fluidity followed by Custom 465, L0H12N4M and 17-4PH. CSS 42L and PH13-8M ranked worst in the fluidity comparison and were therefore excluded from further investigations. Mechanical testing at both ambient and elevated temperatures (400 degrees Celsius) was performed. It was concluded that JETHETE 152M was the best performing alloy with respect to mechanical properties, followed by L0H12N4M, 17-4PH and Custom 465. However, JETHETE 152M was later excluded due to its poor corrosion properties. In the corrosion test, at 400 degrees Celsius, for 100 hours with salt spray fog, it was determined that the 17-4PH and L0H12N4M showed similar corrosion rates. Wettability test performed on two different shell systems with 17-4PH showed that the shell/alloy system is important to consider during filling of thin sections. The 17-4PH alloy was used in the casting trials of a demonstrator. It was demonstrated during the casting trials that filling of a wide flat section with a thickness below 2 mm is hard to achieve

The pathogenesis of endometriosis: Molecular and cell biology insights

The etiopathogenesis of endometriosis is a multifactorial process resulting in a heterogeneous disease. Considering that endometriosis etiology and pathogenesis are still far from being fully elucidated, the current review aims to offer a comprehensive summary of the available evidence. We performed a narrative review synthesizing the findings of the English literature retrieved from computerized databases from inception to June 2019, using the Medical Subject Headings (MeSH) unique ID term \u201cEndometriosis\u201d (ID:D004715) with \u201cEtiology\u201d (ID:Q000209), \u201cImmunology\u201d (ID:Q000276), \u201cGenetics\u201d (ID:D005823) and \u201cEpigenesis, Genetic\u201d (ID:D044127). Endometriosis may origin from M\ufcllerian or non-M\ufcllerian stem cells including those from the endometrial basal layer, M\ufcllerian remnants, bone marrow, or the peritoneum. The innate ability of endometrial stem cells to regenerate cyclically seems to play a key role, as well as the dysregulated hormonal pathways. The presence of such cells in the peritoneal cavity and what leads to the development of endometriosis is a complex process with a large number of interconnected factors, potentially both inherited and acquired. Genetic predisposition is complex and related to the combined action of several genes with limited influence. The epigenetic mechanisms control many of the processes involved in the immunologic, immunohistochemical, histological, and biological aberrations that characterize the eutopic and ectopic endometrium in affected patients. However, what triggers such alterations is not clear and may be both genetically and epigenetically inherited, or it may be acquired by the particular combination of several elements such as the persistent peritoneal menstrual reflux as well as exogenous factors. The heterogeneity of endometriosis and the different contexts in which it develops suggest that a single etiopathogenetic model is not sufficient to explain its complex pathobiology

A Study Using Sulfur K-Edge XAS of Bitumens, Asphaltenes, Maltenes and their Oxidation Products by Comparison with Model Compounds

High resolution Sulfur K-edge XANES spectra have been recorded at the ESRF on "test" samples of bitumen, asphaltene and maltene fractions and various model compounds. All these fractions were found to have a low content of sulfidic species as opposed to thiophenic species and it is therefore not surprising that thermally oxidized samples (RTFOT) had also a low content (1%) of sulfoxides. This is indeed consistent with the absence of any detectable S=0 signature in the EXAFS spectra which were recorded over more than 1300 eV using the "undulator gap scan" technique

New roles for old friends: Involvement of the innate immune system in tumor progression

The identification of various innate immune system effectors in the TME has prompted researchers to investigate their role in tumor progression and metastasis, leading to the development of potential targeted therapies. Undoubtedly, this field is in constant development and much remains to be unraveled. Therefore, we encourage researchers to share their latest findings in this promising field related to the management of oncologic disorders