Improved mass spectrometric proteomic profiling of the secretome of rat vascular endothelial cells

Serum albumin contamination of cells cultured in vitro significantly impedes the mass spectrometric analysis of proteins secreted by the cells. Here we report a novel washing and culturing technique for rat vascular endothelial cells that considerably reduces the concentration of the commonly used additive for cell culture, bovine serum albumin (BSA), in the secretome of these cells. Cells are rinsed stringently and cultured for 24 h in serum-free media without appreciably impeding cell growth or viability. The percentage of BSA scans identified by tandem mass spectrometry (LC-MS/MS) in stringently rinsed cells (average 13.2%) was significantly lower than either the moderately rinsed or no rinse cell treatments (average 35.2% and 45.2% respectively). Furthermore, the stringent wash treatment allowed the confident identification of a larger portion of the secretome of rat endothelial cells by LC-MS/MS.     

Countrywide Survey for MERS-Coronavirus Antibodies in Dromedaries and Humans in Pakistan

Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050)samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to detect IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged [ 10 years (81.37%) followed by those aged 3.1–10 years (78.65%) and B 3 years (58.19%).Higher prevalence was observed in female (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 2016–2017. Among the sampled population, 840 humans were camel herders.Although we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels.     

Sequential abundant ion fragmentation analysis (SAIFA): an alternative approach for phosphopeptide identification using an ion trap mass spectrometer

Phosphorylation has been the most studied of all the posttranslational modifications of proteins. Mass spectrometry has emerged as a powerful tool for phosphomapping on proteins/peptides. Collision-induced dissociation (CID) of phosphopeptides leads to the loss of phosphoric or metaphosphoric acid as a neutral molecule, giving an intense neutral loss product ion in the mass spectrum. Dissociation of the neutral loss product ion identifies peptide sequence. This method of data-dependent constant neutral loss (DDNL) scanning analysis has been commonly used for mapping phosphopeptides. However, preferential losses of groups other than phosphate are frequently observed during CID of phosphopeptides. Ions that result from such losses are not identified during DDNL analysis due to predetermined scanning for phosphate loss. In this study, we describe an alternative approach for improved identification of phosphopeptides by sequential abundant ion fragmentation analysis (SAIFA). In this approach, there is no predetermined neutral loss molecule, thereby undergoing sequential fragmentation of abundant peak, irrespective of the moiety lost during CID. In addition to improved phosphomapping, the method increases the sequence coverage of the proteins identified, thereby increasing the confidence of protein identification. To the best of our knowledge, this is the first report to use SAIFA for phosphopeptide identification.     

P2RY1/ALK3-Expressing Cells within the Adult Human Exocrine Pancreas Are BMP-7 Expandable and Exhibit Progenitor-like Characteristics

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印度地鼠的性成熟和窝仔数

为有效地进行有害脊椎动物的控制 ,研究了栖息于巴基斯坦Punjab中部的 348只印度地鼠的繁殖模式。在所捕捉到的 10 7雄鼠和 2 4 1只雌鼠中 ,分别有 75只和 179只是性成熟的个体。性比偏雌。使用被捕个体标本的体重和体长作为指标 ,发现最小的性成熟的雄性个体体重为 70 - 89g ,体长 13 1- 14 0cm ;一窝仔里面的平均胚胎数为 2 74± 0 15 (范围 ,1- 5 ) ,而平均胎斑数为 4 2 9± 0 19(1- 11) .体重似乎对窝仔数有显著影响 ,而体长和估计的年龄对窝仔数没影响。当老鼠繁殖不活跃和不繁殖时 ,控制害兽最有效     

A new high-elevation scorpion species of the genus Scorpiops Peters, 1861 (Scorpiones: Euscorpiidae: Scorpiopinae) from the Himalayas,India

A new high-elevation scorpion species of the genus Scorpiops is described from the Indian state of Himachal Pradesh. Scorpiops spitiensis sp. nov. is the second highest-elevation scorpion species in Asia and the first one from India occurring at elevations above 4200 m. The new species closely resembles Scorpiops petersii, but it can be distinguished from it based on a suit of characters, one of which is the presence of 16 trichobothria on the external aspect of the patella, which is unique to the new species.     

Cadmium induced oxidative stress and biochemical responses in cyanobacterium <Emphasis Type="Italic">Nostoc muscorum</Emphasis>

The present study deals with the growth, photosynthesis, oxidative stress and heavy metal accumulation ability of Nostoc muscorum exposed to different levels (2, 4, 8, 16, 20 μM) of cadmium (Cd) concentrations. Growth and photosynthetic pigments i.e., chlorophyll a, carotenoids and phycocyanin were significantly affected by cadmium exposure and inhibition was found to be dose dependent. ¹⁴C-fixation appeared to be more sensitive to Cd than whole cell oxygen evolution. Significant accumulation of Cd in the cells of N. muscorum was noticed after 1 and 2 h of exposure and the accumulation rate was dose and time dependent. Furthermore, the levels of superoxide radicals and hydrogen peroxide (H₂O₂) were found significantly increased by cadmium exposure which in turn accelerated the formation of malondialdehyde (MDA) content, and protein and DNA damage. The selected dose of Cd (20 μM) showed the induction of new polypeptide of ~23.24 kD and the loss of ~37.84 kD and ~69.63 kD whereas the remaining bands were inhibited as compared to control. Significant DNA fragmentation which is a hallmark of programmed cell death (PCD) was also observed in the cells treated with 20 μM of Cd for 48 h. The decrease in proline and total phenol content at 8 and 16 μM suggest that the cells of N. muscorum were not able to mitigate the oxidative stress induced by cadmium exposure. Similarly, the decreased activities of antioxidant enzymes i.e., superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) also indicates the failure of the antioxidant defense system of N. muscorum to survive at higher concentration (8 and 16 μM) of cadmium.     

Plasma free amino acid kinetics in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) using a bolus injection of <Superscript>15</Superscript>N-labeled amino acids

To gain insight into the metabolic design of the amino acid carrier systems in fish, we injected a bolus of ¹⁵N amino acids into the dorsal aorta in mature rainbow trout (Oncorhynchus mykiss). The plasma kinetic parameters including concentration, pool size, rate of disappearance (R _d), half-life and turnover rate were determined for 15 amino acids. When corrected for metabolic rate, the R _d values obtained for trout for most amino acids were largely comparable to human values, with the exception of glutamine (which was lower) and threonine (which was higher). R _d values ranged from 0.9 μmol 100 g⁻¹ h⁻¹ (lysine) to 22.1 μmol 100 g⁻¹ h⁻¹ (threonine) with most values falling between 2 and 6 μmol 100 g⁻¹ h⁻¹. There was a significant correlation between R _d and the molar proportion of amino acids in rainbow trout whole body protein hydrolysate. Other kinetic parameters did not correlate significantly with whole body amino acid composition. This indicates that an important design feature of the plasma-free amino acids system involves proportional delivery of amino acids to tissues for protein synthesis.     

Association of nitrogen-fixing,plant-growth-promoting rhizobacteria (PGPR) with kallar grass and rice

Leptochloa fusca (L.) Kunth (kallar grass) has previously been found to exhibit high rates of nitrogen fixation. A series of experiments to determine the level of biological nitrogen fixation using N isotopic dilution were carried out in nutrient solution and saline soil. These studies indicated an agronomically significant amount of nitrogen being fixed in soil. Kallar grass has a similar growth habitat to rice. Therefore similar studies were carried out with rice after isolating various diazotrophs from the roots which were also screened for their ability to produce auxin (IAA). Five such strains namely Azospirillum lipoferum N-4, Azospirillum brasilense Wb-3, Azoarcus K-1, Pseudomonas 96-51, Zoogloea Ky-1 were selected for inoculating two rice varieties i.e. NIAB-6 and BAS-370 under aseptic laboratory conditions. The nitrogen fixed was quantified using the N isotopic dilution method. Variety BAS-370 had nearly 70% nitrogen derived from atmosphere (Ndfa) when inoculated with Azospirillum N-4. Similar studies with the mixed inoculum using N fertilizer in the micro plots indicated that nearly 29% of plant nitrogen was derived from the atmosphere.     

LAP2 Proteins Chaperone GLI1 Movement between the Lamina and Chromatin to Regulate Transcription

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