N-acetylcysteine reduces oxidative stress in sickle cell patients

Oxidative stress is of importance in the pathophysiology of sickle cell disease (SCD). In this open label randomized pilot study the effects of oral N-acetylcysteine (NAC) on phosphatidylserine (PS) expression as marker of cellular oxidative damage (primary end point), and markers of hemolysis, coagulation and endothelial activation and NAC tolerability (secondary end points) were studied. Eleven consecutive patients (ten homozygous [HbSS] sickle cell patients, one HbSβ0-thalassemia patient) were randomly assigned to treatment with either 1,200 or 2,400 mg NAC daily during 6 weeks. The data indicate an increment in whole blood glutathione levels and a decrease in erythrocyte outer membrane phosphatidylserine exposure, plasma levels of advanced glycation end-products (AGEs) and cell-free hemoglobin after 6 weeks of NAC treatment in both dose groups. One patient did not tolerate the 2,400 mg dose and continued with the 1,200 mg dose. During the study period, none of the patients experienced painful crises or other significant SCD or NAC related complications. These data indicate that N-acetylcysteine treatment of sickle cell patients may reduce SCD related oxidative stress

C-Terminus Glycans with Critical Functional Role in the Maturation of Secretory Glycoproteins

The N-glycans of membrane glycoproteins are mainly exposed to the extracellular space. Human tyrosinase is a transmembrane glycoprotein with six or seven bulky N-glycans exposed towards the lumen of subcellular organelles. The central active site region of human tyrosinase is modeled here within less than 2.5 Å accuracy starting from Streptomyces castaneoglobisporus tyrosinase. The model accounts for the last five C-terminus glycosylation sites of which four are occupied and indicates that these cluster in two pairs - one in close vicinity to the active site and the other on the opposite side. We have analyzed and compared the roles of all tyrosinase N-glycans during tyrosinase processing with a special focus on the proximal to the active site N-glycans, s6:N337 and s7:N371, versus s3:N161 and s4:N230 which decorate the opposite side of the domain. To this end, we have constructed mutants of human tyrosinase in which its seven N-glycosylation sites were deleted. Ablation of the s6:N337 and s7:N371 sites arrests the post-translational productive folding process resulting in terminally misfolded mutants subjected to degradation through the mannosidase driven ERAD pathway. In contrast, single mutants of the other five N-glycans located either opposite to the active site or into the N-terminus Cys1 extension of tyrosinase are temperature-sensitive mutants and recover enzymatic activity at the permissive temperature of 31°C. Sites s3 and s4 display selective calreticulin binding properties. The C-terminus sites s7 and s6 are critical for the endoplasmic reticulum retention and intracellular disposal. Results herein suggest that individual N-glycan location is critical for the stability, regional folding control and secretion of human tyrosinase and explains some tyrosinase gene missense mutations associated with oculocutaneous albinism type I

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

An Overview On Development Of Three Dimensional Reinforcements For Use In Composites At CSIR-NAL

3D composites have potential applications in aerospace, space and defence sectors. Skin- stiffener elements in wings, aircraft and spacecraft root fittings, armors, helmets & bullet proof vests, engine components such as nozzles, exhaust cones, convergent-divergent flaps, etc.. are the potential areas of interest for 3D composites. These composites have the ability to improve the out-of-plane properties which has been one of the challenges currently faced by composite designers. The interweaves of the 3D reinforcements in the composite will hold the structure together when subjected to thermal shocks and exposure to high temperatures. Also, it has been shown that the stress analysis conducted on a 3D composite has isotropic thermo-mechanical loading regime. This paper details about the development of 3D weaving technologies for use in composites keeping the above applications in hindsight. The focus of the work is on single layer profile weaving for inserts, angle interlock and noobing technologies for integrated thick structures and 3D weaving technology based on dual-direction shedding principle for integrated thick profiles used in fittings

Evaluation of Antioxidant and Anti-amylase Activities of Sukhasarak Churna, an Ayurvedic Formulation

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Agglutinated foraminifera (superfamily Hormosinacea) across the Indian margin oxygen minimum zone (Arabian Sea)

We present a semi-quantitative survey of ‘live’ (stained) and dead hormosinacean foraminifera at six sites (500–2,000 m water depth; bottom-water oxygen concentrations 0.007–2.43 ml L−1) across the Indian margin oxygen minimum zone (OMZ). Abundance of stained and dead specimens was highest at 800 m followed by 1,100 m, lowest at 2,000 m (stained) and 500 m (dead). The peak at 800 m possibly represents a release from oxygen stress combined with a rich food supply (‘edge effect’). We recognised 31 species (27 Reophax, 2 Hormosinella, 1 Hormosina and 1 Nodosinella) among the 605 stained and dead specimens; the majority (21) are apparently undescribed. Species richness was low at 2,000 m; within the OMZ, it was maximal at 1,100 m and minimal at 500 m for both stained and dead populations. Three species (R. agglutinatus, R. aff. bilocularis and R. dentaliniformis) occurred across the entire depth range. However, most species were either confined to the 2,000-m site or to one or more sites within the OMZ. Multivariate analysis of assemblage composition revealed that the 2,000-m site was distinct from shallower sites. Within the OMZ, the 900- and 1,100-m sites were the most similar, and the 500-m site the most distinct. Stained:dead test ratios were maximal at 500–835 m, perhaps reflecting enhanced preservation of cytoplasm at very low oxygen concentrations. At least two Reophax species are common to the Indian and Pakistan margin OMZ; one of these may be confined to the core of the Arabian Sea OMZ