Analysis of the effect of potato carboxypeptidase inhibitor pro-sequence on the folding of the mature protein.

Protein folding can be modulated in vivo by many factors. While chaperones act as folding catalysts and show broad substrate specificity, some pro-peptides specifically facilitate the folding of the mature protein to which they are bound. Potato carboxypeptidase inhibitor (PCI), a 39-residue protein carboxypeptidase inhibitor, is synthesized in vivo as a precursor protein that includes a 27-residue N-terminal and a seven-residue C-terminal pro-regions. In this work the disulfide-coupled folding of mature PCI in vitro has been compared with that of the same protein extended with either the N-terminal pro-sequence (ProNtPCI) or both N- and C-terminal pro-sequences (ProPCI), and also with the N-terminal pro-sequence in trans (ProNt + PCI). No significant differences can be observed in the folding kinetics or efficiencies of all these molecules. In addition, in vivo folding studies in Escherichia coli have been performed using wild-type PCI and three PCI mutant forms with and without the N-terminal pro-sequence, the mutations had been previously reported to affect folding of the PCI mature form. The extent to which the 'native-like' form was secreted to the media by each construction was not affected by the presence of the N-terminal pro-sequence. These results indicate that PCI does not depend on the N-terminal pro-sequence for its folding in both, in vitro and in vivo in E. coli. However, structural analysis by spectroscopy, hydrogen exchange and limited proteolysis by mass spectrometry, indicate the capability of such N-terminal pro-sequence to fold within the precursor form.     

Intraoperative electron radiation therapy after salvage surgery in gynecological cancers and retroperitoneal sarcomas: outcomes and adverse effects

BackgroundSalvage surgery is considered an option for isolated recurrences of retroperitoneal and pelvic tumors, in patients who have undergone previous radiotherapy. In order to increase local control intra operative electron radiation therapy (IOERT) can be used in these patients to administer additional radiation dose. We evaluated the outcomes and adverse effects in patients with retroperitoneal sarcoma and gynecologic tumors after salvage surgery and IOERT.Materials and methodsTwenty patients were retrospectively analyzed. Twenty-three IOERT treatments were performed after surgery. Six (30%) were sarcoma and 14 (70%) were gynecological carcinoma. Administered dose depended on previous dose received with external beam radiotherapy (EBRT) and proximity to critical structures. The toxicities were scored using the Common Terminology Criteria for Adverse Events version 4.0.ResultsThe median age of the patients was 51 years (range 34–70). After a median follow-up of 32 months (range 1–68), in the sarcoma group the local control rate was 66.6%; while in the gynecological group the local control rate was 64.3%. In relation to late toxicity, one patient had a Grade 2 vesicovaginal fistula, and one patient presented Grade 4 enterocolitis and enteric intestinal fistula.ConclusionsIOERT could have a role in the treatment of retroperitoneal sarcomas in primary tumors after EBRT, as it may suggest a benefit in local control or recurrences after surgical resection in those at high risk of microscopic residual disease. The addition of IOERT to salvage resection for isolated recurrence of gynecologic cancers suggest favorable local control in cases with concern for residual microscopic disease.     

Key Elements in a Framework for Land Use Impact Assessment Within LCA (11 pp)

Background, Aim and Scope Land use by agriculture, forestry, mining, house-building or industry leads to substantial impacts, particularly on biodiversity and on soil quality as a supplier of life support functions. Unfortunately there is no widely accepted assessment method so far for land use impacts. This paper presents an attempt, within the UNEP-SETAC Life Cycle Initiative, to provide a framework for the Life Cycle Impact Assessment (LCIA) of land use. Materials and Methods: This framework builds from previous documents, particularly the SETAC book on LCIA (Lindeijer et al. 2002), developing essential issues such as the reference for occupation impacts; the impact pathways to be included in the analysis; the units of measure in the impact mechanism (land use interventions to impacts); the ways to deal with impacts in the future; and bio-geographical differentiation. Results: The paper describes the selected impact pathways, linking the land use elementary flows (occupation; transformation) and parameters (intensity) registered in the inventory (LCI) to the midpoint impact indicators and to the relevant damage categories (natural environment and natural resources). An impact occurs when the land properties are modified (transformation) and also when the current man-made properties are maintained (occupation). Discussion: The size of impact is the difference between the effect on land quality from the studied case of land use and a suitable reference land use on the same area (dynamic reference situation). The impact depends not only on the type of land use (including coverage and intensity) but is also heavily influenced by the bio-geographical conditions of the area. The time lag between the land use intervention and the impact may be large; thus land use impacts should be calculated over a reasonable time period after the actual land use finishes, at least until a new steady state in land quality is reached. Conclusions: Guidance is provided on the definition of the dynamic reference situation and on methods and time frame to assess the impacts occurring after the actual land use. Including the occupation impacts acknowledges that humans are not the sole users of land. Recommendations and Perspectives: The main damages affected by land use that should be considered by any method to assess land use impacts in LCIA are: biodiversity (existence value); biotic production potential (including soil fertility and use value of biodiversity); ecological soil quality (including life support functions of soil other than biotic production potential). Bio-geographical differentiation is required for land use impacts, because the same intervention may have different consequences depending on the sensitivity and inherent land quality of the environment where it occurs. For the moment, an indication of how such task could be done and likely bio-geographical parameters to be considered are suggested. The recommendation of indicators for the suggested impact categories is a matter of future research.     

Effect of gestational and postnatal environmental temperature on metabolic rate in the altricial rodent, Phyllotis darwini

1. In the altricial rodent, Phyllotis darwini, we found higher body temperatures and faster developmental rates of the thermoregulatory capacity in neonates born from cold- than warm-acclimated mothers.

2. This difference could be explained by maternal effects on the litter, such as high levels of catecholamines and thyroxin levels, high concentration of the uncoupled protein and larger quantity of brown adipose tissue as a consequence of cold acclimation.

3. The exposition of mothers and the maintenance of cold condition during the early development might be responsible of the high metabolism and better thermoregulatory capacity of newborns.

Genetic and morphological differentiation between the two largest breeding colonies of Audouin's Gull Larus audouinii

We assessed the genetic and morphological differences between the two largest breeding colonies of Audouin's Gull Larus audouinii , an endemic seabird species of the Mediterranean region. The two colonies comprise c. 75% of the total world population and are 655 km apart. The Ebro Delta colony was formed recently and, after dramatic growth mainly due to high rates of both immigration and reproductive success, is now the largest in the world (more than 60% of the total population). The Chafarinas Islands support an ancient colony with relatively little fluctuation in breeding numbers. The two colonies also differ greatly in environmental conditions, with the Ebro Delta being a higher quality breeding site. Very little movement occurs between the two colonies. We collected morphological data and blood samples from both colonies. Polymorphic microsatellite markers were used to study the genetic differentiation. These showed no significant variation between colonies, nor evidence of a founder effect in the Ebro Delta. Individuals from the Ebro Delta were larger than those from Chafarinas, the difference being greater for males. This probably reflects a stronger male susceptibility to worse environmental conditions during chick growth at the Chafarinas Islands.     

Gruffi: an algorithm for computational removal of stressed cells from brain organoid transcriptomic datasets

Organoids enable in vitro modeling of complex developmental processes and disease pathologies. Like most 3D cultures, organoids lack sufficient oxygen supply and therefore experience cellular stress. These negative effects are particularly prominent in complex models, such as brain organoids, and can affect lineage commitment. Here, we analyze brain organoid and fetal single‐cell RNA sequencing (scRNAseq) data from published and new datasets, totaling about 190,000 cells. We identify a unique stress signature in the data from all organoid samples, but not in fetal samples. We demonstrate that cell stress is limited to a defined subpopulation of cells that is unique to organoids and does not affect neuronal specification or maturation. We have developed a computational algorithm, Gruffi, which uses granular functional filtering to identify and remove stressed cells from any organoid scRNAseq dataset in an unbiased manner. We validated our method using six additional datasets from different organoid protocols and early brains, and show its usefulness to other organoid systems including retinal organoids. Our data show that the adverse effects of cell stress can be corrected by bioinformatic analysis for improved delineation of developmental trajectories and resemblance to in vivo data.     

Transmembrane adaptor protein WBP1L regulates CXCR4 signalling and murine haematopoiesis

WW domain binding protein 1‐like (WBP1L), also known as outcome predictor of acute leukaemia 1 (OPAL1), is a transmembrane adaptor protein, expression of which correlates with ETV6‐RUNX1 (t(12;21)(p13;q22)) translocation and favourable prognosis in childhood leukaemia. It has a broad expression pattern in haematopoietic and in non‐haematopoietic cells. However, its physiological function has been unknown. Here, we show that WBP1L negatively regulates signalling through a critical chemokine receptor CXCR4 in multiple leucocyte subsets and cell lines. We also show that WBP1L interacts with NEDD4‐family ubiquitin ligases and regulates CXCR4 ubiquitination and expression. Moreover, analysis of Wbp1l‐deficient mice revealed alterations in B cell development and enhanced efficiency of bone marrow cell transplantation. Collectively, our data show that WBP1L is a novel regulator of CXCR4 signalling and haematopoiesis.     

Using life cycle assessment to achieve a circular economy

The International Journal of Life Cycle Assessment - The current global interest in circular economy (CE) opens an opportunity to make society’s consumption and production patterns more...     

Vitamin D receptor BsmI polymorphism modulates soy intake and 25-hydroxyvitamin D supplementation benefits in cardiovascular disease risk factors profile

Vitamin D receptor polymorphisms may predispose that not all individuals could have benefits from the nutritional supplementation of 25-hydroxyvitamin D. Furthermore, vitamin D-related cardiovascular effects may also be influenced by soy isoflavones considered endocrine regulators of cardiovascular homeostasis. To find possible gene–diet interactions by evaluating individualized lipid metabolism benefits from an increase in soy and 25-hydroxyvitamin D intake, 106 healthy individuals, genotyped for vitamin D receptor (VDR) gene polymorphism rs1544410 (BsmI) were randomly assigned to either no intake, to daily 250?mL or 500?mL of a 25-hydroxyvitamin D supplemented SB for 2 months. The soybean beverage induced differences in cardiovascular risk factors (lipid profile, blood pressure, TNFα and MCP-1), as well as vitamin D metabolites in a dose-gene-dependent relation. Thus, VDR BsmI polymorphism affected individual response being the GG genotype the ones that showed dose-dependent manner responsiveness in the reduction in total cholesterol, LDL and triglycerides in comparison with the AA/AG genotype. These differences were associated with increased plasma levels of 1α,25-dyhydroxyvitamin D3 in the carriers of the GG genotype. It was concluded that metabolic response to 25-hydroxyvitamin D and soybean supplementation is dependent on VDR BsmI GG genotype due to a higher conversion rate from vitamin D precursors.