Adenovirus type D and type E infection in broiler chickens: the effect on CD4 and CD8 T cell response, cytokines expression and their immunopathology.

1. A total of 150-day-old chicks were divided into three groups of 50 birds (G1-G3); G1 and G2 were orally inoculated at 1-day old with 0.5 ml of 107 TCID50/ml FAdV-D serotype 2 (MT386509.1) and FAdV-E serotype 8a (MW847902), respectively, and G3 was blank control group.2. Cell-mediated immune response was evaluated by detection of CD4, CD8 T lymphocytes and the mRNA expression of IL6 and IL8 in the chicken spleen using q-PCR. Additionally, immunopathology was performed at 3, 5 and 7 day post infection (dpi) and weekly until the end of the experiment.3. Results revealed that transcription of inflammatory cytokines (IL6, IL8) was up regulated in the spleen of FAdV type D and type E infected chickens at various time points relative to the control group. A marked decrease in the number of CD4 and CD8 T lymphocytes at 5 and 7 dpi in G1 of chickens infected with FAdV type D. Whereas, in chickens infected with FAdV type E, the CD4 and CD8 T lymphocytes were markedly decreased at 7 dpi.4. In contrast, there were no significant differences in humoral immune responses against NDV vaccine in (G1 and G2) at different intervals post-vaccination compared to the control group. The histopathology of the bursa, thymus, and spleen in the infected groups showed lymphocytolysis with severe reticular cells hyperplasia and lymphoid depletion.5. In conclusion, fowl adenovirus types D and E have an immunosuppressive effect in broilers which may be considered one of the main causes of the continuous co-infections with other viruses reported in the field during the last 10 years.

Venlafaxine Determination in Pharmaceutical Formulation and Serum by Ion-Selective Electrodes.

BACKGROUND: The application of an ion selective technique for the determination of analyte concentrations is considered one of the most economical techniques for quality control purposes. OBJECTIVE: To elaborate and investigate the construction and general performance characteristics of potentiometric PVC membrane sensors for venlafaxine cation (Ven+). METHOD: The sensors are based on the use of the ion association complexes of the venlafaxine cation with phosphotungstate (PT) and silicotungstate (ST) counter anions as ion exchange sites in the plasticized PVC matrix. They are characterized by potentiometric and conductimetric measurements, performed under various conditions. RESULTS: The electrodes showed a fast (response time around 15 s), stable (life span 45 days) and linear (r2 0.995) response for venlafaxine over the concentration range of 5x10-5 - 1x10-2 M venlafaxine hydrochloride. The solubility product of the ion pair and the formation of the precipitation reaction leading to the ion pair, were determined conductimetrically. The electrodes were found to be very selective, precise (RSD < 1%) and applicable to the potentiometric determination of venlafaxine hydrochloride in pure solutions or in pharmaceutical preparation and in biological fluid (serum), without any interference. Validation of the method shows the suitability of the proposed electrodes for use in the quality assessment of venlafaxine hydrochloride. CONCLUSION: Using only a pH meter in combination with the selective electrodes, drug substance or drug product could be determined accurately in a few seconds. In addition, the in-house made electrodes were tested to monitor venlafaxine in serum. Acceptable results were achieved using the standard addition technique.

Protective efficacy of a prime-boost protocol using H5-DNA plasmid as prime and inactivated H5N2 vaccine as the booster against the Egyptian avian influenza challenge virus.

In this study, a recombinant DNA plasmid was constructed, encoding for HA1 of a selected Egyptian H5N1 virus (isolated during the 2012 outbreaks). In the immunization and challenge experiments, SPF chickens received 1 or 2 doses of H5-DNA plasmid prime, and boosted with the inactivated H5N2 vaccine. Haemagglutination inhibition (HI) titers, protection levels, and the magnitude of virus shedding were compared to that of the chickens that received either DNA plasmid or inactivated H5N2 vaccine alone. H5N1 virus A/chicken/Egypt/128s/2012 (H5N1) highly pathogenic avian influenza (HPAI) clade 2.2.1/C was used for the challenge. Chickens immunized with 1 or 2 doses of H5-DNA vaccine failed to overcome the challenge with 0% and 10% protection, respectively. Quantitative real-time reverse transcription-PCR revealed virus shedding of 2.2 x 104 PCR copies/ml 3 days post challenge (dpc) in the only surviving bird from the group that received 2 doses of plasmid. However, chickens immunized with 1 or 2 doses of H5-DNA plasmid as prime and inactivated H5N2 vaccine as booster, showed 80% protection after challenge, with a viral shedding of 1.2 x 104 PCR copies/ml (1 dose) and 1.6 x 104 PCR copies/ml (2 doses) 3 dpc. The surviving birds in both groups did not shed the virus at 5 and 7 dpc. In H5N2-vaccinated chickens, protection levels were 70% with relatively high virus shedding (1.8 x 104 PCR copies/ml) 3 dpc. HI titers were protective to the surviving chickens. This study reports the efficacy of H5-DNA plasmid to augment reduction in viral shedding and to provide better protection when applied in a prime-boost program with the inactivated AI vaccine.

Next generation sequencing (NGS): a golden tool in forensic toolkit.

The DNA analysis is a cornerstone in contemporary forensic sciences. DNA sequencing technologies are powerful tools that enrich molecular sciences in the past based on Sanger sequencing and continue to glowing these sciences based on Next generation sequencing (NGS). Next generation sequencing has excellent potential to flourish and increase the molecular applications in forensic sciences by jumping over the pitfalls of the conventional method of sequencing. The main advantages of NGS compared to conventional method that it utilizes simultaneously a large number of genetic markers with high-resolution of genetic data. These advantages will help in solving several challenges such as mixture analysis and dealing with minute degraded samples. Based on these new technologies, many markers could be examined to get important biological data such as age, geographical origins, tissue type determination, external visible traits and monozygotic twins identification. It also could get data related to microbes, insects, plants and soil which are of great medico-legal importance. Despite the dozens of forensic research involving NGS, there are requirements before using this technology routinely in forensic cases. Thus, there is a great need to more studies that address robustness of these techniques. Therefore, this work highlights the applications of forensic sciences in the era of massively parallel sequencing.

Echinacea purpurea and Allium sativum as immunostimulants in fish culture using Nile tilapia (Oreochromis niloticus).

The study was conducted to evaluate the efficiency of echinacea (E) and garlic (G) supplemented diets as immunostimulant for tilapia (Oreochromis niloticus). Seven treatments were designed including a control (C). Fish were fed on 35% protein diet at a rate of 3% body weight per day. Echinacea (1.0 ppt) and garlic (3%) were incorporated in the feed, which was administered for periods of 1, 2 and 3 months (summer season), followed by basal diet for 4 more months (winter season). Neutrophil adherence and haematocrit values increased in both supplemented groups with prolonging period of application. The neutrophils adherence was significantly increased in all treatments except group administered echinacea for 1 month. The lymphocytic counts were significantly (p < 0.004) elevated that resulted in a significant increase in the total leucocytic count in groups administered echinacea for 1 and 2 months when compared with the control and/or other treatments. The gain in the body weight and specific growth rate was significantly increased in all supplemented groups (p < 0.004) during summer, but remained without any significant increase after winter. The survival rate was significantly high (>85%) in all the supplemented groups. The percentage of protection, after challenge infection using pathogenic Aeromonas hydrophila was the highest in groups supplemented with echinacea and garlic for 3 months after summer and winter seasons. It could be concluded that echinacea and garlic improve the gain in body weight, survival rate and resistance against challenge infection. Both compounds showed extended effects after withdrawal and improved resistance to cold stress during the winter season. However, a full commercial cost benefit analysis is necessary before recommending their application in aquaculture.

Study of growth in prepubertal asthmatics.

OBJECTIVE: The aim of this pilot study was to assess whether long standing asthma affects growth in prepubertal Egyptian children before initiation of long-term corticosteroid therapy. METHODS: Children with asthma were divided into two groups according to asthma severity, moderate (n=24) and severe (n=14) and were compared for their physical and skeletal growth with a control group (n=15) using standard deviation score (SDS) and one-way ANOVA (analysis of variance) test. RESULTS: No statistically significant differences were found between various growth parameters (weight, height, BMI, upper segment lower segment ratio, and skin fold thickness in asthmatic and normal children, although a positive correlation was found between the age at which the asthma presented and the height in all asthmatic children, r= 0.288, p= 0.036. The bone age standard deviation scores (SDS) were 0.97 mean, -0.165 and -0.572 for controls, moderate and severe asthmatics respectively (P< 0.05), and significant inter group difference between the 2 asthmatic groups (moderate and severe) and the controls was found. CONCLUSION: The authors conclude that there was no significant major effect of asthma per se on growth parameters in children, but that skeletal maturation was influenced by long standing asthma.

Determination of meloxicam in bulk and pharmaceutical formulations.

Three sensitive and reproducible methods for quantitative determination of meloxicam (mel) in pure form and in pharmaceutical formulations are presented. The first method is high performance liquid chromatography by which the drug is determined in the presence of its degradation products over concentration range 100-500 microg x ml(-1) with mean percentage accuracy 100.13+/-0.53. The second method is based on measuring the absorbance of the formed neutral complex between basic methylene blue and mel in phosphate buffer (pH 8) at lambda=653.5 nm over concentration range 1-5 microg x ml(-1) with mean percentage accuracy 99.12+/-1.18. The third method is based on reaction between 2,3-dichloro-5,6-dicyano-p-benzoquinone resulting in the formation of an intense orange red coloured product after heating in a boiling water bath for 5 min. The coloured product exhibits an absorption maximum at 455 nm, over concentration range 40-160 microg x ml(-1) with mean percentage accuracy 100.53+/-1.04.

Determination of aceclofenac in bulk and pharmaceutical formulations.

Three sensitive and reproducible methods for quantitative determination of aceclofenac (AC) in pure form and in pharmaceutical formulation are presented. The first method is based on the reaction between the drug via its secondary aromatic amino group and p-dimethylaminocinnamaldehyde (PDAC) in acidified methanol to give a stable coloured complex after heating at 75 degrees C for 20 min. Absorption measurements were carried out at 665.5 nm. Beer's law is obeyed over concentration range 20-100 microg ml(-1) with mean recovery 100.33 +/- 0.84. The other two methods are high performance liquid chromatography (HPLC) and densitometric methods by which the drug was determined in the presence of its degradation products over concentration range of 20-70 microg ml(-1) and 1-10 microg per spot and mean recoveries are 99.59 +/- 0.90 and 99.45 +/- 1.09, respectively.

Detection of coproantigen in early trichinellosis.

Trichinellosis has become undoubtedly worldwide in distribution. Its diagnosis relies largely on the serodiagnostic procedures which are of great value but unfortunately miss the enteric phase. This could be a serious diagnostic problem in the absence of corresponding epidemiological data and typical symptoms and signs of the disease. In this study the possibility of coproantigen detection, as an early diagnostic aid in trichinellosis, was investigated in mice experimentally infected with Trichinella spiralis. A modified double sandwich ELISA was developed using polyclonal antibodies raised in rabbits and guinea pigs against larval somatic antigens. The first detection of coproantigen was as early as the first day post infection, gradually increasing to reach its peak on the seventh day and then decreasing to disappear completely on the third week post infection. Another test, the coagglutination test (Co-A) was used, and this test confirmed the previous results. The finding of this study suggest that the coproantigen detection could be exploited to confirm ongoing early Trichinella spiralis infection. This fast and easy to use diagnostic method should improve the early infection in human.

Effects of Nigella sativa oil on gastric secretion and ethanol induced ulcer in rats.

The present work was done to investigate the possible effects of Nigella sativa oil (NSO) on gastric secretion and ethanol-induced ulcer in rats. Thirty two adult male rats were used in this study (four groups) and several parameters were determined to assess any degree of protection. It was found that the administration of NSO in rats produced a significant increase in mucin content and glutathione level and a significant decrease in mucosal histamine content. Ethanol administration produced a 100% ulcer induction with an ulcer score of 12.62+/-1.35 (mean+/-S.E., n=8). It caused a significant reduction in free acidity and glutathione level while it produced a significant increase in mucosal histamine content. When animals were pretreated with NSO before induction of ulcer, there was a significant increase in glutathione level, mucin content and free acidity and a significant decrease in gastric mucosal histamine content with a protection ratio of 53.56% as compared to the ethanol group. It can be concluded that NSO imparted a protective action against ethanol induced ulcer in rats.

DNA analysis of hepatocytes: a novel method for estimating the value of liver tissue vaccine (LTV).

Because of the socio-economic condition of African countries, the traditional types of vaccines should be encouraged. LTV in a preliminary study, proved to be of value in reducing morbidity and mortality in Schistosoma mansoni infection. In the present study and because of the vaccine contains liver tissue homogonate, the present authors analyed the DNA content of the hepatocytes of the vaccinated and control groups of animals to know any effects of the vaccine on the hepatocytes ploidy. The DNA content of vaccinated animals was higher. The results were discussed. It was concluded that for testing S. mansoni vaccine, DNA analysis of heptocytes for detecting proliferative S-phase is an important as a positive immune reaction.

Heated fats. Part 3. Biological effect and effect of heating and tempering oils on fatty acid composition of liver, heart and serum lipids of rats.

The study deals with the biological effect caused by changes in fats during heating. The study includes feeding experiments and extraction of serum, liver, and heart from the animals tested. The biological study reveals that animals fed heated oil showed retardation of growth, poor efficiencies, rough, greasy mottled coats, and shortened life span.