Expression of the novel human gene,UBE2Q1, in breast tumors

The novel human gene, designated ubiquitin-conjugating enzyme E2Q family member 1 (UBE2Q1) maps to chromosome 1q21.3. The gene has an open reading frame corresponding to 422 amino acids and contains a RWD domain and an E2 ubiquitin conjugating enzyme domain. Here, we investigated the expression levels of both mRNA and protein of UBE2Q1 gene in cancerous versus normal parts of breast specimens from 26 patients. Real-time PCR data showed that the relative expression level of UBE2Q1 mRNA was significantly greater in cancers than in non-cancerous tissues of breast specimens (Mean ± SEM, 0.064 ± 0.015 for cancers and 0.026 ± 0.01 for noncancerous tissues, P < 0.05 Mann–Whitney test). A rabbit polyclonal antibody was generated against an amino acid sequence predicted from the DNA sequence of UBE2Q1 gene. This antibody was used to perform Western blotting on 21 cases in our cohort of breast specimens. Thus, 13 (61.904%) of the cases showed an increase in the UBE2Q1 immunoreactivity in their cancerous tissues as compared with the corresponding normal tissues. This result along with the real-time PCR data shows that the novel human gene, UBE2Q1, is expressed in human breast and may have implications for pathogenesis of breast cancer.     

Serum concentrations of MCP-1 and IL-6 in combination predict the presence of coronary artery disease and mortality in subjects undergoing coronary angiography

The UBA–UBX domain-containing proteins can interact with ubiquitinated substrates and p97 during endoplasmic reticulum-associated degradation (ERAD). Here, we found that the expressions of all UBA–UBX genes p47, SAKS1, UBXD8, FAF1, and UBXD7 were elevated upon ER stress, albeit with different levels. Of which p47, SAKS1, and UBXD8 are ‘immediate’ respondents whereas FAF1 and UBXD7 were ‘late’ respondents to ER stress. Interestingly, the expression of specific UBA–UBX genes were altered in cells stably expressing three different ERAD substrates such as α-TCR, α1-antitrypsin, and δCD3. We first found that p47 and UBXD8 expression levels were increased in α-TCR and α1-antitrypsin stable cell lines, respectively, whereas SAKS1 expression level was reduced in all the three ERAD substrates tested. Of note, we also found p47 promotes, whereas SASK1 delays the degradation of a single ERAD substrate, α-TCR. Additionally, we found that SAKS1 selectively inhibits the degradation of ERAD substrates without affecting cytosolic proteasomal substrates. Taken together, our results identified that UBA–UBX proteins possess substrate selectivity and opposite role of two different UBA–UBX proteins in the degradation of a single ERAD substrate.     

UBE2Q1 expression in human colorectal tumors and cell lines

Colorectal cancer is the third most common cancer in the world. Ubiquitin–proteasome system has shown to be activated in colorectal and other malignancies. UBE2Q1 is a novel human gene that encodes a putative E2 ubiquitin conjugating enzyme. Here, we investigated the expression pattern of UBE2Q1 gene in cell lines and tissues from human colorectal tumors. Quantitative (q) RT-PCR were employed to evaluate the expression levels of the mRNA for UBE2Q1 in colorectal cancer cell lines (HT29/219, LS180, SW742, Caco2, HTC116, SW48, SW480 and SW1116). Expression of UBE2Q1 at the protein levels were assessed by Western blotting in cell lines as well as in 43 human colorectal tumor tissues. All cell lines tested expressed UBE2Q1 gene at the level of both mRNA and protein, with the SW1116 line representing the lowest level of expression. The cell lines HT29/219 and SW742 showed the highest levels of UBE2Q1 protein and mRNA respectively. When compared to corresponding normal tissues, malignant parts of colorectal tumors showed increased levels of UBE2Q1 immunoreactivity in 32 (74.42 %) of cases. These data suggest that UBE2Q1 is differentially expressed in colorectal cell lines and shows overexpression in colorectal tumors.     

Cloning and overexpression of Bacillus cereus penicillin-binding protein 3 gene in Escherichia coli

The pbp3 gene encoding PBP3 of Bacillus cereus was cloned and sequenced. For this purpose, PBP3 was first purified from B. cereus ts-4, and N-terminal amino acid sequences of the peptides obtained from the protease digests of the protein were analyzed. The B. cereus ts-4 pbp3 gene consisted of an open reading frame of 1,986 bp encoding 662 amino acid residues with a calculated molecular mass of 73,044 Da. The active site-motifs SXXK, SXN, and KTG are present at the positions 393, 452, and 590, respectively, in the deduced amino acid sequence. The pbp3 structural gene was ligated into the pET17 x b expression vector and pET-pbp3 was constructed. A protein was produced by the cells of E. coli carrying pET-pbp3. The produced protein migrated at about 75 kDa in SDS-polyacrylamide gel and strongly reacted with biotinylated ampicillin.     

Investigating the role of potassium and urea to control fruit drop and to improve fruit quality of “Dhakki” date palm

Fruit drop is a key issue with date palm cultivars that can be addressed with a variety of methods and strategies. Foliar application of macronutrients can be more effective in inhibiting fruit drop and improving the quality of date fruits. The current study was carried out to investigate the possible role of potassium (K) and urea to reduce fruit drop and improve the fruit quality of “Dhakki” date palm. It was conducted in a complete randomised block design with seven treatments and three replications at Pakistan''s Agricultural Research Institute, Dera Ismail Khan. The treatments used were: (i) Control (distilled water spray); (ii) Potassium sulphate (K₂SO₄) at 1 %; (iii) K₂SO₄ at 1 % + Urea at 2 %; (iv) K₂SO₄ at 2 %; (v) K₂SO₄ at 2 % + Urea at 2 %; (vi) K₂SO₄ at 3 % and; (vii) K₂SO₄ at 3 % + Urea at 2 %. All the concentrations were sprayed at Kimri stage of fruit development during two consecutive growing seasons. Twenty-one date palms of equal size and age were chosen for the assessments to measure percent fruit drop and other physicochemical variables, including fruit length, fruit diameter, fruit weight, pulp percentage, yield/bundle, pH, total soluble solids (TSS), K content in fruit, and all sugars (percent) of harvested date fruit. The results revealed that bunch spray of K significantly affected all the parameters during both seasons. Application of K₂SO₄ alone and in combination with urea not only effectively reduced the fruit drop but also improved fruit quality in date where, K₂SO₄ applied at 2 % combined with urea was the best concentration in reducing fruit drop, enhancing other physicochemical attributes, and improving fruit quality of “Dhakki” date palm. This study may effectively contribute to reduce the fruit drop and enhance the fruit quality by using K and urea, enabling farmers to improve the date yield and increase economic growth.     

Global dynamic modes of peristaltic-ciliary flow of a Phan–Thien–Tanner hybrid nanofluid model

Biomechanics and Modeling in Mechanobiology - In this paper, the tools of dynamical systems theory are applied to examine the streamline patterns and their local and global bifurcations for...     

Efficacy of dietary supplemental insoluble fibrous materials in ameliorating adverse effects of coccidial challenge in broiler chickens

ABSTRACT

The current experiment was designed to examine effects of dietary supplemental sunflower hulls (SH) and rice hulls (RH) on growth performance, carcass traits, intestinal morphology, lesion score and oocyst shedding in broiler chickens exposed to coccidial challenge. A total of 540 broiler chickens (Ross 308) were assigned to six dietary treatments based on a factorial arrangement (2 × 3) across 1–14, 14–28 and 28-42-d periods. Experimental treatments consisted of broiler chickens without or with coccidial challenge each offered with three different diets: a basal diet or basal diet supplemented with either RH or SH at 40 g/kg diet, respectively. Infection with Eimeria impaired daily weight gain (DWG) and feed conversion ratio (FCR) of broiler chickens during growing period (p < 0.05) while supplementation of SH or RH reduced the adverse effect of coccidiosis so that birds had similar DWG to those fed the basal diet without infection. However, only dietary SH improved the FCR of broilers challenged with coccidiosis. Regardless of coccidial challenge, dietary access to insoluble fibre improved performance of broilers across the growing period (p < 0.05); however, this effect was not observed during the entire rearing period. Relative weights of liver and pancreas were increased in birds subjected to coccidial challenge on d 21 of age (p < 0.05). Moreover, relative weights of the intestinal segments were enhanced (p < 0.05). Furthermore, gizzard weights were higher in birds receiving diets added with fibre (p < 0.05). Infection with coccidiosis decreased villus height and villus height to crypt depth ratio in duodenum of broilers which received the basal diet compared with those fed the same feed without coccidial challenge (p < 0.05). However, supplemental SH could decrease the negative effect of infection on the noted intestinal morphometric attributes. Similarly, a marked reduction was observed for lesion score and faecal oocyst excretion of challenged broilers fed on dietary supplemental fibre (p < 0.05). In conclusion, supplementation of insoluble fibre could ameliorate negative effects of coccidial challenge on DWG of broiler chickens and inclusion of SH in diet of birds exposed to Eimeria infection could be recommended.     

Effect of vehicle on intrauterine absorption of gentamicin in cattle

An intrauterine (IU) infusion of 4 ml gentamicin solution (G) was administered in a water vehicle to 4 normal cycling and healthy cows and in a saline vehicle to 4 other cows during the diestrous phase (days 7–14) of the cycle. Infusion solution (IS) and blood samples were collected periodically postinfusion. G concentration was determined in plasma and IS by radioimmunoassay. Eighty-three and 18% of the infused G disappeared from the uterine lumen by 6 hours postinfusion on using water and saline vehicles, respectively.Total amounts of G reaching the blood stream were nearly the same during the 6 hours postinfusion period using water and saline vehicles. This suggested the possibility of retention of drug in uterine tissue using the distilled water vehicle.     

Effects of glucobrassicin, epiprogoitrin and related breakdown products on locusts feeding: Schouwia purpurea and desert locust relationships

The glucosinolates of a Saharan crucifer Schouwia purpurea (Forskål) (Brassicaceae) were determined by liquid chromatography. Two of these glucosinolates and sinigrin were tested for their deterrent effect on Schistocerca gregaria (Forskål) (Orthoptera: Acrididae). Glucobrassicin, three indolyls and epigoitrin were synthesized for this purpose. Epiprogoitrin was extracted from Crambe seeds. Choice tests on artificial substrate compared feeding responses to glucosinolates and to related breakdown products released when the plant is eaten. Breakdown products were more efficient in deterring the generalist locust than were glucosinolates. Two patterns of dose responses were recorded: glucosidic compounds deterred or stimulated feeding, depending on the concentration tested; aglycones did not stimulate feeding at any concentration. Allyl isothiocyanate, a volatile compound, was a 100-fold higher deterrent than its substrate (sinigrin).     

Thermolysis of derivatives of amino sugars

Amino groups influence the course of the pyrolysis of carbohydrate derivatives and result in substantial dehydration and charring. N-Phenylglycosylamines and their acetates rearrange with almost quantitative loss of water or acetic acid and retention of the arylamino groups. The sharp dehydration reaction contrasts with the behaviour of the corresponding phenyl glycosides on thermolysis, which results in cleavage of the phenolic groups and subsequent breakdown of the glycosyl moiety. Thermal analysis and chemical data indicate that charring of other derivatives of amino sugars is due to intermediate formation of glycosylamines from thermal reaction of the amino group with glycosidic centres.