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Cloning of Vibrio cholerae outer membrane protein W in Pichia pastoris.

BACKGROUND AND OBJECTIVE: The outer membrane protein W (ompW) of Vibrio cholerae is involved in stimulating the immune response via inductio … sistant transformants, clones were selected and subsequently confirmed for cloning by PCR enzymatic digestion and sequencing. RESULTS: PCR, enzymatic digestion and sequencing showed that the ompW gene was correctly cloned into P. pastoris genome. CONCLUSION: Results of our study showed that the methylotrophic yeast P. pastoris can be considered as an appr …

Protein quantification of UDP-glucuronosyltransferases 1A1 and 2B7 in human liver microsomes by LC-MS/MS and correlation with glucuronidation activities.

The aims of this study were to quantify absolute protein levels of uridine 5'-diphosphate-glucuronosyltransferases (UGTs) 1A1 and 2B7 in hum … vine serum albumin was used as a blank matrix for calibration curve samples. Our procedure had good digestion efficiency, sensitivity, calibration curve linearity, and reproducibility of digestion to quantification. In 16 individual HLMs, the protein levels of UGT1A1 and UGT2B7 ranged from 6.50 to 44.6 pmol/mg and 4.45 to 18.2 pmol/mg, respectively. Estradi …

Key gene regulatory sequences with distinctive ontological signatures associate with differentially endonuclease-accessible mouse sperm chromatin.

Using a well-established endonuclease-based chromatin dissection procedure in conjunction with both experimental comparative genome hybridisation (CGH) array profiling and in silico data mining, we show that mouse spermatozoa contain chromatin that is sensitive and resistant to digestion with micrococcal nuclease (MNase). Sequences represented in the micrococcal nuclease digestion solubilised (MNDS) but not the MND insoluble (MNDI) chromatin are strongly enriched in chromosomal regions of high gene density. Furthermore, by fluorescence in situ hybridisation (FISH) analysis, we show that MNDS a…

Differential DNA methylation as a tool for noninvasive prenatal diagnosis (NIPD) of X chromosome aneuploidies.

The demographic tendency in industrial countries to delay childbearing, coupled with the maternal age effect in common chromosomal aneuploidies and the risk to the fetus of invasive prenatal diagnosis, are potent drivers for the development of strategies for noninvasive prenatal diagnosis. One breakthrough has been the discovery of differentially methylated cell-free fetal DNA in the maternal circulation. We describe novel bisulfite conversion- and methylation-sensitive enzyme digestion DNA methylation-related approaches that we used to diagnose Turner syndrome from first trimester samples. We…

[Construction of the recombinant secretion type BCG-Eg95 vaccine of Echinococcus granulosus].

OBJECTIVE: To construct the recombinant secretion type BCG-Eg95 vaccine of Echinococcus granulosus (rsBCG-Eg95). METHODS: BCG-Ag85B signal sequence with 117 bp and Eg95 gene with 471 bp were amplified from the genome of BCG and pGEX-4T-Eg95 by PCR, respectively. BCG-Ag85B signal coding gene and Eg95 gene were cloned into E. coli-BCG shuttle-vector pMV261 to get the recombinant plasmid pSMEg95, which was confirmed by restriction endonuclease digestion, PCR amplification and gene sequencing. These recombinant plasmids were introduced into BCG by electroporation for the construction of rsBCG-Eg9…

[Detection of two novel mutations of iduronate-2-sulfatase gene in patients with mucopolysaccharidosis type II].

In the present study, through PCR amplification and direct sequencing of mutation "hotspots", we were able to identify two novel mutations i … es from normal individuals, indicating that they were not polymorphisms. The PCR-restriction enzyme digestion showed that the two newly identified mutations were of maternal origin, which was consistent with the X-linked recessive disorder. These findings suggest that the IDS gene mutations could be detected by amplifying mutation "hotspots", direct sequenc …

Повышение иммуноактивности с хелаторами в целях повышения чувствительности к обнаружению человека prPSc за счет западного пятна.

Прионские заболевания представляют собой нейродегенеративные расстройства, затрагивающие людей, как болезнь Креуцфельдта-Якоба. Закодированный белок приона (PRP(C)) будет преобразован в структурно изоформу (PRP(Sc)). PrP(Sc) различаются по размерам и гликоформам и могут быть идентифицированы с помощью молекулярного печатания с западной пятнистостью. Электрофоретическая мобильность ПР(Sc) изменяется при обработке металлическими ионами или хелаторами до пищеварения выступами. В ходе этого исследования мы изучили последствия применения хелаторов к пр(Sc) после того, как переваривание протеза не б

Совпадение в аннотации оригинала
Prion diseases are neurodegenerative disorders affecting humans as Creutzfeldt-Jakob disease. The host-encoded prion protein (PrP(C)) will b … The electrophoretic mobility of PrP(Sc) changes on treatment with metal ions or chelators prior to digestion with proteases. The effects of chelators applied to PrP(Sc) after protease digestion had not been examined in detail, we investigated these effects in this study. Application of EDTA, NTA and DTPA, and to a lesser extent EGTA, significantly enhanced …

Relation between the outer cover of the egg case of Argiope aurantia (Araneae: Araneidae) and the emergence of its spiderlings.

To emerge from the egg case, Argiope aurantia spiderlings must penetrate a tightly woven outer cover composed primarily of large-diameter cylindrical gland fibers and small-diameter fibers, likely of aciniform gland origin. They accomplish this using enzymatic digestion and mastication to form a communal hole in the outer cover. The involvement of proteolytic enzymes in this process was demonstrated by zymography of spiderling homogenates and washes made from the edges of holes. The specific source(s) of the proteases is unknown, but histological examination of spiderling sections indicates th…

[Histochemical studies of glycosaminoglycans in the developing eyelids of experimental microphthalmic (Cts) mice].

PURPOSE: The Cts mouse is a mutant strain which induces congenital cataract and small eyes in homozygotes. The process of eyelid development in Cts mice was examined histochemically. MATERIALS AND METHODS: Prenatal (from day 14 of gestation) and postnatal (to day 14) animals of unaffected and homozygous mice were examined. Sensitized high iron diamine staining was done after chondroitinase B/testicular hyaluronidase double digestion, and sensitized low iron diamine staining was done after Streptomyces hyaluronidase digestion. RESULTS: The main chondroitin sulfate isomer in the dermis of the …

Oral colonization is unlikely to play an important role in Helicobacter pylori infection.

BACKGROUND: This clinical and microbiological study investigated whether Helicobacter pylori colonizes the mouth en route to the pyloric ant … d histological examination. Gastric and duodenal juice samples were aspirated. Restriction nuclease digestion with HaeIII was employed on all specimens from patients in whom there was evidence of the organism in the mouth. RESULTS: H. pylori was observed in dental plaque in only 15 patients, all from the 116 who had evidence of the organism in the stomach. …

Ultrastructural distribution of sulfated glycosaminoglycans in epithelial-mesenchymal interface of developing rat tooth germs.

We investigated the ultrastructural distribution of sulfated glycosaminoglycans in the epithelial-mesenchymal interface of tooth germs by use of the high-iron diamine thiocarbohydrazide silver proteinate (HID-TCH-SP) staining and enzymatic digestion method. At an early stage in odontoblast differentiation, HID-TCH-SP stain deposits were sparsely distributed in the basement membrane and in the intercellular spaces. Subsequently, as formation of the initial predentin matrix began, HID-TCH-SP stain deposits were densely distributed in the interfibrillar spaces and the basement membrane. Testicula…

Contributions of ascariasis to poor nutritional status in children from Chiriqui Province, Republic of Panama.

Relationships between ascariasis and lactose digestion and between ascariasis and food transit time from mouth to caecum were investigated in young children from Chiriqui Province, Republic of Panama. The breath hydrogen method was used in both studies. Ascaris-infected children showed a significantly poorer degree of lactose digestion following a test oral load than uninfected children. Recovery of the capacity of the children to digest lactose was still not fully complete for at least 3 weeks following anthelmintic treatment. On average, the mouth-to-caecum transit time was similar in infect…

Structural differences between liver- and muscle-derived insulin receptors in rats.

The structure of insulin receptors, solubilized from rat skeletal muscle and liver, was studied. The alpha subunit was identified by specifi … lated beta subunits migrated with a Mr of 98,000 for muscle and 101,000 for liver. After partial V8 digestion of autophosphorylated, immunoprecipitated receptors the major phosphopeptide fragment migrated on SDS-PAGE at Mr 57,000 from muscle and 60,000 from liver. Glycosidase digestion of autophosphorylated receptors suggested that Mr heterogeneity was due …

Is gastric sham feeding really sham feeding?

Rats were fitted with gastric cannulas, food deprived, and allowed to drink a sugar solution that drained out of the opened cannula; i.e., the rats sham-fed. Although this procedure is thought to prevent absorption of ingested food, it was found that the sham feeding of a 32% glucose or sucrose solution significantly elevated blood glucose levels. The addition of acarbose, a drug that inhibits the digestion of sucrose, to the 32% sucrose solution blocked the blood glucose rise, as did closing the pylorus with an inflatable pyloric cuff. Neither the drug nor the cuff, however, reduced the amoun…

Effectiveness of second-grade Trigonella foenum-graecum seed supplementation on voluntary intake, nutrient utilization, and growth performance in Black Bengal goats.

BACKGROUND: Fenugreek (Trigonella foenum-graecum) seeds contain bioactive compounds that may enhance nutrient utilization and growth in small ruminants. However, fenugreek is often expensive and may compete with human food requirements. AIMS: This study evaluated the effect of second-grade fenugreek seed supplementation with concentrate ration on intake, nutrient digestibility, and growth performance of Black Bengal goat kids. METHODS: A total of 18 post-weaned Black Bengal female kids were selected and divided into three groups. The control group was fed a basal diet (a mixture of roughage

Energy and phosphorus evaluation of poultry meal fed to broiler chickens using a regression method.

Two experiments were conducted to determine energy (Exp. 1) and P (Exp. 2) utilization in poultry meal (PM) for broiler chickens. A total of 192 birds were allotted to 3 experimental diets in a randomized complete block design with BW as a blocking factor on d 15 and 16 post hatching in Exp. 1 and 2, respectively. Each diet was fed to 8 replicate cages with 8 birds per cage in both experiments. Initial BW of birds in Exp. 1 and 2 were 438 ± 76.9 g and 543 ± 50.2 g, respectively. Three corn-soybean meal-based diets were prepared to contain 0, 80, or 160 g/kg in Exp. 1 and 0, 50, or 100 g/kg in

Breadfruit flour is a healthy option for modern foods and food security.

Breadfruit is a traditional staple crop from Pacific islands with the potential to improve worldwide food security and mitigate diabetes. Flour produced from breadfruit is a gluten-free, low glycemic index, nutrient dense and complete protein option for modern foods but basic scientific knowledge of health impacts of a breadfruit-based diet in animals and humans was lacking. We designed a series of studies to provide basic and fundamental data on impacts of a breadfruit-based diet through an in vitro and in vivo model. Cooked breadfruit flour was digested through a multi-stage enzyme digestion…

Characterization of Clostridium Perfringens Isolates Collected from Three Agricultural Biogas Plants over a One-Year Period.

Digestate produced by agricultural biogas plants (BGPs) may contain pathogenic bacteria. Among them, Clostridium perfringens deserves particular attention due to its ability to grow under anaerobic conditions and persist in amended soil. The aim of this study was to examine the potential pathogenicity and the antimicrobial resistance of C. perfringens in manure and digestate collected from three agricultural biogas plants (BGPs). A total of 157 isolates (92 from manure, 65 from digestate) were screened for genes encoding seven toxins (cpa, cpb, etx, iapcpe, netB, and cpb2). The 138 cpa positiv

Aorto-mesenteric space reduction in women with anorexia nervosa: retrospective audit and analysis.

BACKGROUND AND AIMS: Superior mesenteric artery syndrome (SMAS) is a rare condition favored by weight loss, with nonspecific digestive complaints that may hinder weight recovery in anorexia nervosa. This study aimed to examine the radiological features of aorto-mesenteric (A-M) space reduction in anorexia nervosa and their association with digestive complaints. METHODS: Female patients with anorexia nervosa and a history of computerized tomography angiography for digestive complaints were included. Clinical data were retrospectively collected and computerized tomography scans were reviewed by

Multimorbidity Patterns and Depression: Bridging Epidemiological Associations with Predictive Analytics for Risk Stratification.

Background: Late-life depression is a critical public health concern, particularly among older adults with chronic multimorbidity. Existing studies often focus on single-disease associations, neglecting the complex interplay of coexisting conditions. Understanding how multimorbidity patterns contribute to depression risk and identifying high-risk subgroups through integrated statistical and machine learning approaches remain underexplored, limiting targeted prevention strategies. Methods: Using data from the China Health and Retirement Longitudinal Study (CHARLS), latent class analysis (LCA) w