Sabry M. El-Bahr

Simultaneous multiresidue determination of metronidazole and spiramycin in fish muscle using high performance liquid chromatography with UV detection

An efficient multiresidue method for the simultaneous determination of metronidazole (MET) and spiramycin (SPY) in tilapia fish muscle, based on high performance liquid chromatography with UV detection (HPLC-UV), has been developed. The drugs were extracted with 0.2% orthophosphoric acid-methanol (6:4), and the extracts were cleaned up on a solid phase extraction cartridge, C18 Sep-Pak light column. The LC separation was performed on a RP stainless-steel C-18 analytical column (150 mm x 4.6 mm, 5 microm) with a gradient elution system of 0.05 M phosphate buffer adjusted to pH 2.4-acetonitrile as the mobile phase at the flow rate of 1.0 ml min(-1). A wavelength programming was applied for the UV detection of the analytes. The method not only enabled the determination of the parent drugs, MET and SPY, but also permitted the determination of their metabolites, hydroxymetronidazole (HMET) and neospiramycin (NSPY). The calibration graphs for each drug were rectilinear in the range of 0.005-1.000 microg g(-1) for MET and HMET and 0.025-1.000 microg g(-1) for SPY and NSPY. With this method, the cited drugs with their metabolites were determined in fortified fish muscle tissues at levels of 0.025, 0.1 and 1.0 microg g(-1) with good accuracy and precision. LOD and LOQ obtained for each drug were as follows: 0.002 and 0.005 microg g(-1) for MET and HMET and 0.005 and 0.025 microg g(-1) for SPY and NSPY. Utilization of the method to successfully analyze tilapia fish muscle samples incurred with MET and SPY was described.

Biochemical and histopathological study in rats intoxicated with carbontetrachloride and treated with camel milk

The unique characters of camel’s milk make it used extensively in the field of medicine as anti-microbial, anti-diabetic and hepatoprotective agent. The lack of studies demonstrating the protective effect of camel’s milk against hepatotoxic compound was the main reason beyond the conduction of the current experiment which aimed to investigate the protective effects of camel’s milk against carbontetrachloride (CCl4) induced hepatotoxicity. Therefore, 24 rats were fed on standard diet and divided into four groups. Rats of the first group and second groups were injected i/p with paraffin oil and received either tap water (control 1) or camel’s milk (control 2), respectively. Rats of the third and fourth groups were injected i/p with CCl4 and received either tap water or camel’s milk, respectively. At the end of the experiment (5 weeks), blood and liver samples were collected for biochemical and histopathological analysis. The present findings revealed that, CCl4 elevated serum enzyme activities of liver and some biochemical parameters, but these effects were prevented by the treatment of rats with camel milk. Histopathologically, a great amount of mononuclear cells infiltration, necrotic cells and few fibroblasts were observed in liver of CCl4 treated group. The present study concluded that camel milk treatment may play a protective role against CCl4-induced liver damages in rats. These protective effects were in the form of improving of liver enzyme activities, blood biochemical parameters and histological picture of liver of intoxicated rats. In the future, examination of the liver protective effect of camel milk against CCl4 in dose dependant manner could be investigated.

Serum biochemical profile and molecular detection of pathogens in semen of infertile male dromedary camels (Camelus dromedarius)

Detection of pathogens in the semen of camels has not been completely elucidated. Therefore, the current study aimed to determine the association of some economically important pathogens with infertility in 94 male infertile camels through molecular detection and estimation of selected biochemical parameters in serum of these animals compared with a control non infected fertile animals (n=40). PCR analysis of semen samples of infertile camels indicated that, four potential pathogens namely Mycoplasma spp., Leptospira spp., Brucella melitensis, and Bovine viral diarrhea virus (BVDV) were detected in 50 semen samples of infertile camels whereas, 44 semen samples of infertile camels were free of pathogens and all tested semen samples were negative for bovine herpes virus 1, Salmonella spp. and Trypanosoma evansi. Single and mixed infection was detected in 88% and 12% of the infected semen samples, respectively. Mycoplasma spp., Leptospira spp., Brucella and Bovine viral diarrhea virus infection represented 66%, 27.2%, 4.5% and 2.3% of the single infected semen samples. Mycoplasma spp.+Leptospira spp. and Mycoplasma spp.+Brucella spp. were detected in 83.3% and 16.7% of mixed infected semen samples, respectively. Testosterone concentration decreased significantly in infertile infected camels compare to both control and infertile non infected animals that remained comparable. The current findings reported the molecular detection of mixed infection in camel semen for the first time. Mycoplasma spp. is the most widely recognized microorganism in the present study and together with Leptospira spp., Brucella spp. and Bovine viral diarrhea virus, might be associated with infertility in dromedary camels.

Biochemical and hormonal analysis of follicular fluid and serum of female dromedary camels (Camelus dromedarius) with different sized ovarian follicles

The current study aimed to compare some biochemical and hormonal constituents in follicular fluids and serum of female dromedary camels with different sized ovarian follicles. Therefore, follicular fluids from follicles sized 1.1-1.5cm (n=10), 1.6-2.1cm (n=10) and 2.2-2.5cm (n=10) and sera were harvested from 20 female camels. The concentrations of ascorbic acid, glucose, cholesterol and activities of acid phosphatase (ACP) and alkaline phosphatase (ALP) were not changed significantly neither in follicular fluids of all follicle sizes nor in sera of female camels with different sized follicles. The concentrations of estradiol-17β (E2) in the follicular fluid of follicles sized 2.2-2.5cm were significantly lower (P<0.01) than its corresponding value in follicular fluid of other follicle sizes. The concentrations of progesterone (P4), tri-iodothyronine (T3), thyroxin (T4), cortisol and insulin-like growth factor-1 (IGF-1) remained comparable in follicular fluids of all examined different sized follicles. The concentrations of E2, P4, T3, T4, cortisol and IGF-1 were similar in the serum of camels with different sized follicles. Interestingly, mean concentrations of P4 and IGF-1 in follicular fluids were higher than their corresponding values in sera of camels with different sized follicles and the mean concentrations of glucose, cholesterol, ALP and cortisol in sera were higher than their corresponding values in follicular fluids of the examined camels. With the exception of E2, there were no significant differences in biochemical and hormonal constituents between follicular fluids from different sized follicles.

Cancer cell death induced by nanomagnetolectin

Magnetic nanoparticles represent a new paradigm for molecular targeting therapy in cancer. However, the transformative targeting potential of magnetic nanoparticles has been stymied by a key obstacle-safe delivery to specified target cells in vivo. As cancer cells grow under nutrient deprivation and hypoxic conditions and decorate cell surface with excessive sialoglycans, sialic acid binding lectins might be suitable for targeting cancer cells in vivo. Here we explore the potential of magnetic nanoparticles functionalized with wheat germ lectin (WGA) conjugate, so-called nanomagnetolectin, as apoptotic targetable agents for prostate cancer. In the presence of magnetic field (magnetofection) for 15min, 2.46nM nanomagnetolectin significantly promoted apoptosis (∼12-fold, p value <0.01) of prostate cancer cells (LNCaP, PC-3, DU-145) compared to normal prostate epithelial cells (PrEC, PNT2, PZ-HPV-7), when supplemented with 10mM sialic acid under nutrient deprived condition. Nanomagnetolectin targets cell-surface glycosylation, particularly sialic acid as nanomagnetolectin induced apoptosis of cancer cells largely diminished (only 2 to 2.5-fold) compared to normal cells. The efficacy of magnetofected nanomagnetolectin was demonstrated in orthotopically xenografted (DU-145) mice, where tumor was not only completely arrested, but also reduced significantly (p value <0.001). This was further corroborated in subcutaneous xenograft model, where nanomagnetolectin in the presence of magnetic field and photothermal heating at ∼42°C induced apoptosis of tumor by ∼4-fold compared to tumor section heated at ∼42°C, but without magnetic field. Taken all together, the study demonstrates, for the first time, the utility of nanomagnetolectin as a potential cancer therapeutic.

Effect of prednisolone on thyroid and gonadotrophic hormones secretion in male domestic rabbits

Context: An excess of endogenous and exogenous glucocorticoids causes many biochemical endocrine effects. Such alterations resulted in erroneous misleading interpretation of laboratory results. Aims: The effect of six intramuscular (I/M) injections of prednisolone on the serum concentrations of thyroxin (T 4 ), triiodothyronine (T 3 ), thyroid stimulating hormone (TSH), and luteinizing hormone (LH) of male domestic rabbits was assessed. Materials and Methods: Ten rabbits were assigned into two equal groups, treated and control groups. The treated group was injected prednisolone 1% at a rate of 2.2 mg/kg body weight every other day for a total of six doses. The control group was injected I/M with an equivalent volume of vehicle solution of prednisolone 1%. Statistical Analysis: All data were presented as mean ± standard error of mean by Student′s t-test. All tests were performed using the Statistical Package for Social Science Program. Results: At day 6 of the experiment (after three injections of prednisolone), there were nonsignificant differences (P > 0.5) in serum T 4 , T 3 and TSH concentrations in prednisolone-treated animals compare to control. However, the concentration of serum LH was decreased significantly (P < 0.5) in prednisolone-treated animals compare to control. At day 12 of the experiment (after six injections of prednisolone), there were nonsignificant changes (P > 0.5) in the concentration of TSH in prednisolone-treated animals compare to control whereas the concentrations of serum T 4 , T 3 and LH were decreased significantly (P < 0.5) in prednisolone-treated animals compare to control. Conclusion: Multiple administration of prednisolone was clearly associated with substantial reduction in the concentrations of T 4 and T 3 and reduction of LH concentration without any effect on TSH concentration in serum of domestic rabbits.

Thermal Manipulation Mid-term Broiler Chicken Embryogenesis: Effect on Muscle Growth Factors and Muscle Marker Genes

ABSTRACT Thermal manipulation (TM) during broiler chicken embryogenesis has been shown to promote muscle development and growth. However, the molecular bases of promoting broiler muscle development and growth are not fully understood. The aim of this study was to investigate the molecular bases of muscle growth and development in broiler chickens subjected to TM. This included the investigating of the changes in mRNA expression levels of muscle marker genes, namely MyoD, myogenin, paired box transcription factor (Pax7) and proliferating cell nuclear antigen (PCNA), and muscle growth factors namely insulin-like growth factor 1 (IGF-1), myostatin and growth hormone (GH) during embryogenesis and on posthatch days 10 and 28. Fertile Cobb eggs (n=1500) were divided into four groups. Eggs in the first group (control) were incubated at 37.8°C and 56% RH, whereas, eggs in the second group (TM 1 ) , third group (TM 2 ) , and fourth group (TM 3 ) were subjected to 39 oC and 65% RH daily during embryonic days (ED) 12–18 for 9, 12, and 18 hours, respectively. Body weight (BW) during embryogenesis and posthatch days (1, 3, 5, 7, 14, 21, 28 and 35) was recorded. mRNA expression levels of muscle marker genes and muscle growth factor genes during ED 12, 14, 16 and 18 and on posthatch days 10 and 28 were analyzed using real-time RT-PCR. TM upregulated the mRNA expressions of muscle marker and growth factors genes. This upregulation was accompanied by improvement of body weight near and at market age.