Heat shock-induced enhanced susceptibility of barley to Bipolaris sorokiniana is associated with elevated ROS production and plant defence-related gene expression.

Heat stress alters plant defence responses to pathogens. Short-term heat shock promotes infections by biotrophic pathogens. However, little is known about how heat shock affects infection by hemibiotrophic pathogens like Bipolaris sorokiniana (teleomorph: Cochliobolus sativus). We assessed the effect of heat shock in B. sorokiniana-susceptible barley (Hordeum vulgare cv. Ingrid) by monitoring leaf spot symptoms, B. sorokiniana biomass, ROS and plant defence-related gene expression following pre-exposure to heat shock. For heat shock, barley plants were kept at 49 °C for 20 s. B. sorokiniana biomass was assessed by qPCR, ROS levels determined by histochemical staining, while gene expression was assayed by RT-qPCR. Heat shock suppressed defence responses of barley to B. sorokiniana, resulting in more severe necrotic symptoms and increased fungal biomass, as compared to untreated plants. Heat shock-induced increased susceptibility was accompanied by significant increases in ROS (superoxide, H2 O2 ). Transient expression of plant defence-related antioxidant genes and a barley programmed cell death inhibitor (HvBI-1) were induced in response to heat shock. However, heat shock followed by B. sorokiniana infection caused further transient increases in expression of HvSOD and HvBI-1 correlated with enhanced susceptibility. Expression of the HvPR-1b gene encoding pathogenesis-related protein-1b increased several fold 24 h after B. sorokiniana infection, however, heat shock further increased transcript levels along with enhanced susceptibility. Heat shock induces enhanced susceptibility of barley to B. sorokiniana, associated with elevated ROS levels and expression of plant defence-related genes encoding antioxidants, a cell death inhibitor, and PR-1b. Our results may contribute to elucidating the influence of heat shock on barley defence responses to hemibiotrophic pathogens.

Signature of real-time PCR in detection of Trichomonas vaginalis infection and its association with human papillomavirus genotype 16.

OBJECTIVE: Infection with Trichomonas vaginalis (TV) is the most prevalent non-viral sexually transmitted infection in the world. The objective of the study was to investigate the incidence of TV infection and its association with Human Papillomavirus (HPV) in a sample of Egyptian females. PATIENTS AND METHODS: 96 Egyptian females suspected for trichomoniasis were involved in our study. Vaginal washouts and cervical (cytobrush) samples were obtained from all patients and examined for T. vaginalis by direct wet mount, modified Diamond's culture medium, and real-time PCR. Cervical (cytobrush) samples were examined for HPV using real-time PCR. RESULTS: Out of 96 patients, 28 (29%) was positive for T. vaginalis by real-time PCR. HPV was detected in 33 patients (34.4%); 31 cases (32.3%) were infected with HPV of genotype 16, whereas only two cases (2.1%) had genotype 18 infection. A significant association was found between TV and HPV infection [Odds ratio (OR)=10.58; 95% confidence interval (CI): 3.819 - 29.29; p<0.001]. CONCLUSIONS: When it comes to diagnosing trichomoniasis in a susceptible population, real-time PCR is more reliable than traditional standard approaches. A significant association between TV and HPV infection was found. Further research into the processes by which these two organisms interact at the cellular level could be revealed.

Effect of date palm (Phoenix dactylifera L.) leaves on productive performance of growing lambs.

Eighteen 4-month-old lambs, with a mean live weight (LW) of 19.47 ± 0.20 kg, were used to evaluate the nutritive value of date palm leaves (DPL) ensiled with different additives in a completely randomized design. Lambs were stratified into three groups of 6 lambs each and fed a control diet comprising 60% concentrate feed mixture (CFM) and 40% DPL silage (T1). In other treatments, the DPL silage (DPLS) of the control treatment was replaced with EM1 additive-treated DPLS (T2) or El-Mofeed additive-treated DPLS (T3). Apparent digestibility, total digestible nutrient, digestible crude protein, dry matter intake, daily weight gain (DWG), price of DWG, daily profit, and economics of feed efficiency were higher (P < 0.05) for the additives-treated DPLS relative to the control, with T2 enhancing these parameters compared with T3. With exception of ruminal pH, which was reduced, concentrations of ruminal NH3-N and total volatile fatty acids (VFA) increased 4 h post feeding. However, ruminal NH3-N and total VFA were greater (P < 0.05) for the additives-treated DPLS, with T2 producing higher values than T3. Ruminal pH and feed cost/kg LW gain were lower for T2 relative to other treatments. Blood constituents were within the normal ranges for lambs, though slightly altered by treatments. Whereas serum total protein, albumin, and globulin were affected (P < 0.05) in this rank order, T1 < T3 < T2, other serum parameters were not affected. Relative feed cost and relative daily profit were lower and higher respectively for T2 than for T3. It is concluded that additives-treated DPLS is nutritionally superior to untreated DPLS as a roughage source in total mixed rations fed to growing lambs. However, for improved performance of the lambs and economic benefits, EM1-treated DPLS is recommended.

Long non-coding RNA EGFR-AS1 in colorectal cancer: potential role in tumorigenesis and survival via miRNA-133b sponge and EGFR/STAT3 axis regulation.

BACKGROUND: Colorectal cancer is one of the most common cancers worldwide and a major cause of cancer-related death. Thus molecular biomarkers for colorectal cancer have been proposed. The role of long non-coding RNA EGFR-AS1 in colorectal cancer is still unclear. We aimed to evaluate its expression in different stages of colorectal cancer and determine any possible role in regulating the miR­133b/EGFR/STAT3 signalling pathway. MATERIALS AND METHODS: The relative expression of EGFR-AS1 and miR­133b were evaluated by quantitative real-time RT-transcription PCR in 130 colorectal cancer samples and 30 normal tissues. EGFR expression was assessed using immunohistochemistry. Furthermore, levels of p-EGFR, p-STAT3, and apoptotic proteins were determined by ELISA. RESULTS: Both EGFR-AS1 and EGFR overexpression were positively linked with colorectal cancer status (both p < 0.01), grade (both p < 0.01), and metastasis (P < 0.01 and p = 0.019 respectively). EGFR-AS1 and miR-133b were significantly inversely correlated (P < 0.01). Low expression of miR-133b was inversely associated with overexpressed EGFR and increased p-STAT3 levels. EGFR-AS1 was an independent prognostic factor for survival of colorectal cancer patients (P < 0.01, HR 2.06; 95% CI 1.32-3.19) where low EGFR-AS1 expression was associated with higher survival rate (p = 0.003). CONCLUSION: EGFR-AS1 may have a role in colorectal cancer by regulation of miR­133b/EGFR/STAT3 signalling. It may be a potential biomarker for early diagnosis and predicting the survival rate of colorectal cancer.

Construction of ovine GH-pmKate2N expression vector and its uptake by ovine spermatozoa using different methods.

This study aims to produce transgenic ovine spermatozoa bearing Ossimi sheep growth hormone (Os_GH) cDNA using different methods. The complete coding sequence of Os_GH has been registered in GenBank accession no. KP221575. The sequence of Os_GH cDNA has been subcloned into pmkate2-N expression vectors to construct Os_GH-pmKate2-N vector. Five groups of sperm uptake were submitted. All groups were incubated at 37 °C for 1 h: Control (sperm cells were incubated without vector), Traditional incubation (sperm cells were incubated with vector), Heat shock (sperm cells were incubated with vector at 4 °C for 20 min and heated for 2 min at 42 °C), Heat shock + Dimethyl sulfoxide (DMSO) (sperm cells were incubated with vector and supplemented with 3% of DMSO and then submitted to heat shock regime) and DMSO (sperm cells were incubated with vector and supplemented with 3% DMSO). The sperm genomic DNA in groups was extracted. The Os_GH-pmKate2-N vector was introduced efficiently into the head of sperm cells in all treated groups. Adding DMSO either with or without heat shock increased the sperm uptake. The progressive motility was reduced (P < 0.05) by 29.9% in heat shock group compared to the control. Adding DMSO improved (P < 0.05) the total and progressive motilities by 8.2% and 19.8%, respectively in heat shock group compared to the heat shock group without DMSO. The results documented the ability of ovine spermatozoa to uptake the exogenous vector. Also, sperm incubation with 3% DMSO is the best method to introduce the exogenous vector into spermatozoa without notable adverse effects on sperm motilities.

Characteristics of ovarian follicular dynamics throughout the estrous cycle of Egyptian buffaloes.

Data of 56 normal and 9 abnormal estrous cycles were collected from 9 Egyptian buffaloes (Bublus bublis) to describe the follicular growth wave pattern. Heat was checked twice daily while, ovaries were scanned daily to monitor the patterns of follicular waves. Day of ovulation was determined when the largest follicle was replaced by corpus haemorrhgicum (CH). Number of waves/cycle, day of emergence of the follicular wave, characteristics of the dominant follicle and corpus luteum (CL) growth features were monitored. Buffaloes displayed mainly two types of follicular waves; two (46.4%) and three (53.6%). In cycles of three wave pattern, time of emergence of the 1st wave post-heat was longer (P<0.05) and number of recruited follicles/wave were larger (P<0.05) compared to the corresponding values of the two wave pattern. Number of recruited follicles in early follicular waves (1st or the 2nd) had larger number (P<0.05) compared to the subsequent ones. Follicles that reached ovulation in both types of estrous cycle had shorter life-span (P<0.05) than the previous ones. Life-span of CH, growing and regressed CL were 3.6+/-0.6, 11.2+/-0.8 and 4.4+/-0.5 days, respectively with no difference in both types of follicular wave. Three types of ovarian disorders were observed. Follicular waves and CL growth features showed unique pattern for each individual. These results demonstrate that buffaloes display two main types of follicular waves with dominance of three wave type.

Suppression of tobacco mosaic virus-induced hypersensitive-type necrotization in tobacco at high temperature is associated with downregulation of NADPH oxidase and superoxide and stimulation of dehydroascorbate reductase.

Tissue necroses and resistance during the hypersensitive response (HR) of tobacco to tobacco mosaic virus (TMV) are overcome at temperatures above 28 degrees C and the virus multiplies to high levels in the originally resistant N-gene expressing plants. We have demonstrated that chemical compounds that generate reactive oxygen species (ROS) or directly applied hydrogen peroxide (H(2)O(2)) are able to induce HR-type necroses in TMV-inoculated Xanthi-nc tobacco even at high temperatures (e.g. 30 degrees C). The amount of superoxide (O(2)(*-)) decreased, while H(2)O(2) slightly increased in TMV- and mock-inoculated leaves at 30 degrees C, as compared with 20 degrees C. Activity of NADPH oxidase and mRNA levels of genes that encode NADPH oxidase and an alternative oxidase, respectively, were significantly lower, while activity of dehydroascorbate reductase was significantly higher at 30 degrees C, as compared with 20 degrees C. It was possible to reverse or suppress the chemically induced HR-type necrotization at 30 degrees C by the application of antioxidants, such as superoxide dismutase and catalase, demonstrating that the development of HR-type necroses indeed depends on a certain level of superoxide and other ROS. Importantly, high TMV levels at 30 degrees C were similar in infected plants, whether the HR-type necrotization developed or not. Suppression of virus multiplication in resistant, HR-producing tobacco at lower temperatures seems to be independent of the appearance of necroses but is associated with temperatures below 28 degrees C.

The effect of dietary menhaden, olive, and coconut oil fed with three levels of vitamin E on plasma and liver lipids and plasma fatty acid composition in rats.

The effect of dietary fats with varying degrees of unsaturation in the presence of different concentrations of vitamin E on tissue lipid levels was studied in rats. Rats were fed either menhaden oil, olive oil or coconut oil at 15% levels with either 0.1, 0.3 or 0.6 mg/g of vitamin E as alpha-tocopherol for four weeks. Rat serum and liver were analyzed for total cholesterol, HDL-cholesterol, triacylglycerol and phospholipids. In addition, fatty acid composition of serum lipids was also analyzed. Serum total cholesterol and triacylglycerol were significantly lower in rats fed menhaden oil than in those fed olive or coconut oil, while the HDL-cholesterol was significantly higher in serum of rats fed menhaden and olive oil than in those fed coconut oil. Levels of vitamin E in the diet had only a significant effect on serum cholesterol and liver phospholipids. The Pearson correlation coefficient showed a significant positive relationship between serum triacylglycerol and total cholesterol, and a negative correlation between triacylglycerol and HDL-cholesterol, and between total and HDL-cholesterol.In the liver, total cholesterol was significantly higher in rats fed coconut oil than in rats fed menhaden oil. Total liver phospholipids were lower in rats fed either coconut oil or olive oil compared to those fed menhaden oil, especially with higher levels of vitamin E intake. Higher levels of vitamin E in the diet appear to increase triacylglycerol and phospholipids in livers of rats fed menhaden oil. In the liver a significant negative correlation was observed between phospholipids and cholesterol. The type and degree of unsaturation (polyunsaturated fatty acids in menhaden oil, monounsaturated fatty acids in olive oil and saturated fatty acids in coconut oil) significantly affected plasma and tissue lipids.

Biochemical effect of antioxidants on lipids and liver function in experimentally-induced liver damage.

Recent studies demonstrated the role of antioxidants in preventing organ damage caused by free radicals. The present study was conducted to find out the modulatory effect of some antioxidants on lipid patterns in experimentally-induced liver damage. Rats chronically intoxicated with carbon tetrachloride (CCl4) were used as a model of liver injury terminating with fibrosis or cirrhosis. One hundred and sixty six albino rats were classified into five groups: one served as a control group; the second was subjected to oral administration of CCl4 (200 microL/100 g body weight) twice a week; the other three groups, in addition to CCl4, received oral doses of silymarin (30 mg/kg), vitamin E (200 IU/kg) and vitamin C (50 mg/kg) respectively. At the end of the experiment, the animals were killed, blood was collected and liver was taken for histopathological examination. Liver function tests, disturbed by CCl4 were significantly modulated by antioxidants, and histopathological examination showed that antioxidants ameliorated the necrotic and fibrotic changes caused by CCl4. Treatment with antioxidants was also shown to modulate the toxic effect of CCl4 on the lipid profile and malondialdehyde content. Administration of antioxidants could play an important role in prophylaxis against lipid peroxidation and consequently liver fibrosis caused by free radicals.

Mycoplasma gallisepticum vaccination-challenge: an egg-production model.

Specific-pathogen-free layer hens in maximum lay were exposed by aerosol to a broth culture of Mycoplasma gallisepticum R' strain. Egg-production loss of greater than 50% was evident 7-14 days following challenge of unvaccinated chickens, with a gradual recovery during the next several days. Various vaccine preparations were tested to determine the effect in the model. All vaccinated chickens exhibited significantly (P < or = 0.05) lower egg-production loss than the unvaccinated controls. The model provides a method for testing treatment effects on egg-production losses resulting from controlled exposure to M. gallisepticum and may be useful in simulating field exposure.

Demonstration of the genetic stability of a Mycoplasma gallisepticum strain following in vivo passage.

A Mycoplasma gallisepticum strain designated 6/85 (MGI) exhibiting reduced virulence for both chickens and turkeys was sequentially passaged 10 times in each species. DNA extracted from organisms before passage and those isolated after the third, sixth, and 10th passages was studied by restriction endonuclease DNA analysis using BamHI, BglII, EcoRI, HindIII, and PstI endonucleases. The virulent-type strain designated S6 was used as a comparison. Comparison of DNA fragment patterns of MGI and S6 strains showed distinct differences, although some similarities were evident. Passage of the strain in vivo did not affect DNA fragment patterns of the MGI strain. Electrophoretic protein patterns produced by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed very similar band patterns in both the MGI and S6 strains. The most notable differences were seen in bands located in the molecular-mass regions of approximately 46.5, 50-54, 58-64, and 105-140 kilodaltons. Alteration of band pattern profiles following in vivo passage of the MGI strain was apparent in a single band at approximately 86 kilodaltons that appeared to stain more intensely following passage.

Evaluation of a Mycoplasma gallisepticum strain exhibiting reduced virulence for prevention and control of poultry mycoplasmosis.

Two experiments were conducted to evaluate the virulence and vaccination efficacy of a Mycoplasma gallisepticum (MG) isolate designated MG Intervet 6/85. Virulence of the strain was determined by evaluation of airsacculitis scores following aerosol exposure to the isolate before and after 10 sequential passes in either commercial broiler chickens or commercial turkeys. Two-week-old specific-pathogen-free chickens were vaccinated by aerosol exposure. The birds were challenged with the R' strain of MG at either 4 or 8 weeks post-vaccination. Efficacy was evaluated by airsacculitis scores determined 21 days after challenge. Ten repetitive back-passes of the isolate in chickens and turkeys did not substantially increase the virulence. Virulence for both chickens and turkeys was minimal, while protection elicited by aerosol vaccination in young chickens against virulent R' strain was significant (P less than or equal to 0.05) compared with unvaccinated controls.

Proposed weaning formula for developing countries.

PIP: This article proposes a process of combining soybean, corn and wheat to make an inexpensive weaning formula, and suggests that animal feeding studies, human organoleptic (taste) tests and finally feeding trials on infants be conducted. The method combines 3 steps involving soybean yogurt, corn starch solution and cracked wheat gruel. First a yogurt is fermented from cracked soybeans cooked in sodium bicarbonate solution, with sucrose added, and homogenized. Then corn is cracked, soaked, boiled, homogenized and sieved. Wheat bulgar (cooked cracked wheat) is powdered and boiled. The soy, corn and wheat components are mixed 3:2:1, sterilized and packaged. These operations would require equipment to mill, cook, time, measure and adjust pH, incubate, homogenize, sieve, weigh, compute dry weight, sterilize and package the formula. The authors tabulate published amino acid analysis of soy, corn and wheat. They propose to test the formula on laboratory animals to determine the feed efficiency ratio compared to standard chow diets, to test organoleptic quality on adult humans and finally to test the formula on weaned infants. It is anticipated that this formula will be inexpensive and appropriate for children with lactose or galactose intolerance or milk allergy.^ieng

Methionine toxicity in chicks and poults.

In feeding experiments with poults, 2% DL-methionine caused a marked growth depression which could be alleviated by the addition of glycine. Homocystine at an equimolar level depressed growth to a lesser degree than methionine, and this growth depression could be alleviated by glycine. Betaine could alleviate the growth depression of homocystine but not that of methionine. Methionine-fed poults developed a cervical paralysis similar to that of a folic acid deficiency, but the addition of this vitamin at several times the requirement was ineffective in counteracting the toxicity of methionine in either chicks or poults.

The effect of diet on the toxicity of vanadium.

Five experiments were conducted with growing chicks to determine the effectiveness of various materials in modifying the toxicity of vanadium. The toxicity of vanadium (as it is measured by growth depression and mortality) was much greater when vanadium was added to a semipurified diet than when it was added to a practical diet containing natural ingredients. When EDTA was added to diets containing 50 to 200 p.p.m. vanadium, the growth depression was reduced from 22.1% and 75.9% to 8.4% and 36.7% respectively, and the mortality was reduced from 80% to 20% among chicks fed 200 p.p.m. vanadium. The addition of 10% lactose to a diet containing 100 p.p.m. vanadium increased the growth depression from 41% to 76.2% and caused 80% mortality. Chromium added to diets containing 100 and 200 p.p.m. vanadium improved the growth and reduced the mortality.

The effect of pharmacological levels of dietary vanadium on the egg production, shell thickness and egg yolk cholesterol in laying hens and coturnix.

The addition of 100 p.p.m. of vanadium to a ration for White Leghorn laying hens caused a significant increase (p less than 0.01) in the egg shell thickness, but did not cause any significant change in the production, egg weight or egg ypok cholesterol content. The addition of 300 p.p.m. of vanadium caused a severe depression in the egg production which was the only symptom of vanadium toxicity to be observed. The addition of 50, 100, 200 or 300 p.p.m. vanadium to the ration of laying coturnix did not cause any significant change in egg production, egg weight, or egg yolk cholesterol content. The addition of the same levels of vanadium to one-day old male coturnix chicks did not cause any significant growth depression or mortality. We suggest that coturnix may be more resistant to vanadium toxicity than chickens.

The effect of dietary vanadium on fatty acid and cholesterol synthesis and turnover in the chick.

Day-old male, broiler type chicks were used to study the effect of 100 ppm dietary vanadium on fatty acid and cholesterol synthesis and turnover in vivo. After feeding the experimental diets for 4 weeks body weight and liver weight of chicks fed 100 ppm vanadium were significantly less than those of the control chicks and liver total lipid and cholesterol tended to be slightly higher than the levels of the control chicks. [1-14C] Acetate was administered intravenously and the specific activities of plasma and liver total lipid, cholesterol and fatty acid were determined at 0.25, 0.50, 1.0, 4.0, 8.0 and 15.0 hours after the injection. Plasma total lipid and cholesterol were significantly higher than the levels in the control chicks. The rate of incorporation of [1-14C]acetate into plasma and liver total lipid, cholesterol and fatty acid was higher in chicks fed vanadium than the control group at any of the time being tested after the injection. There was a significant increase in the hepatic citrate cleavage enzyme activity among chicks fed 100 ppm vanadium, whereas, there was no significant change in acetate thiokinase activity. Turnover rate of plasma total lipid and fatty acid in vanadium fed chicks was lower than the control. The turnover rate of plasma cholesterol determined by administering [4-14C]cholesterol and periodically measuring the specific activity of plasma cholesterol was higher in chicks fed vanadium than in those fed the basal diet.