The potential gonadoprotective effects of grape seed extract against the histopathological alterations elicited in an animal model of cadmium-induced testicular toxicity.

BACKGROUND: Grape seed extract (GSE) is a powerful antioxidant containing high levels of bioflavonoids, vitamin C and vitamin E. The aim of the work is to study the possible protective and ameliorative effects of grape seed extract in an animal model of cadmium (Cd)-induced testicular toxicity in rats. MATERIALS AND METHODS: A thirty-day oral gavage study in adult male albino rats was performed using 32 animals, randomly divided into four equal groups; negative control, Cd (5 mg/k/day), GSE (100 mg/k/day), and Cd + GSE. Testicular weights were measured. Haematoxylin and eosin (H&E) staining and proliferating nuclear cell antigen (PCNA) immunohistochemistry, as a marker for proliferation were done. Morphometric parameters were assessed and subjected to statistical analysis. RESULTS: The H&E results showed atrophy and distortion of the seminiferous tubules (STs) with sloughing of the spermatogenic epithelium in cadmium group. The interstitial spaces were widened and showed oedema and mononuclear cell infiltrations. No remarkable changes were observed in the GSE-only group when compared to the control group. In Cd + GSE group, maintaining of the STs and their lining cells was evident. The immunohistochemical results showed marked positive PCNA immunoreactivity in both control and GSE groups, while negative immunoreaction was noticed in Cd group. Limited positive PCNA immunoreactivity was ameliorated in Cd + GSE group. CONCLUSIONS: Grape seed extract protected against cadmium-induced testicular toxicity in rats, reducing induced histopathological changes, and maintaining testicular histoarchitecture.

A simple real-time polymerase chain reaction assay using SYBR Green for hepatitis C virus genotyping.

Detection of the hepatitis C virus (HCV) genome is crucial for diagnosis of HCV infection and for monitoring the efficacy of HCV treatment. Thus, we aimed to develop a convenient screening test for common HCV genotypes based on melting curve analysis with PCR. Serum samples were drawn from 124 patients with known HCV infection confirmed to be antibody and HCV RNA positive. A characteristic melting curve was obtained by monitoring the fluorescence as the temperature increased through the melting point of the PCR product. Results were compared with those obtained by the restriction fragment length polymorphism (RFLP) genotyping method. The melting curve analysis indicated that the different genotypes had discrete melting points (P < 0.0001): 90.43 +/- 0.065 degrees for genotype 1 (n = 35), 90.21 +/- 0.064 degrees for genotype 2 (n = 18), 90.62 +/- 0.045 degrees for genotype 3 (n = 29) and 90.84 +/- 0.130 degrees for genotype 4 (n = 42). The genotype was determined for all samples using the newly developed method as well as RFLP, and the two systems produced concordant results. The sensitivity of the assay was 91.4 % for genotype 1, 83.3 % for genotype 2, 93.1 % for genotype 3, and 85.7 % for genotype 4. Genotypes detected by melting curve analysis significantly correlated with those detected by RFLP (r = 0.946, P < 0.0001) with a strong linear relationship (r 2 = 0.895). This melting curve analysis is a rapid, convenient and low-cost screening test for differentiation of HCV genotypes 1-4.

Occurrence of iris yellow spot virus from onion crops in Egypt.

Iris yellow spot virus (IYSV) is a infects onion bulb and seed crops in many countries including Egypt. Results of the mechanical inoculation reveled that, small chlorotic lesions and systemic necrosis were observed on both Nicotiana benthamiana and Datura stramonium after 10 days, while there were no symptoms were appeared on the onion plant. The viral biological transmission with Thrips tabaci was highly reported to be efficiently for virus transmitted. Our results confirmed the presentence of virus-like particles of a Tospovirus infected onion leaf using transmission electron microscopy. Both of sequence and phylogenetic analysis of N gene revealed that our viral isolate is IYSV with 95 % identity with reported Israel isolate. The sequence of N gene had three motifs: casein kinase II Phosphorylation site, N-myristoylation site and protein kinase C phosphorylation site. These motifs are involved in regulation, activity and stability of IYSV. To our knowledge, this is the first molecular characterization of IYSV in Egypt.

First Report of Iris yellow spot virus Infection of Garlic and Egyptian Leek in Egypt.

Egyptian leek (Allium ampeloprasum), garlic (A. sativum), and onion (A. cepa) are key vegetables produced by small- and large-scale farmers in Egypt for national and international markets. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an economically important viral pathogen of bulb and seed onion crops in many onion-growing areas of the world (1,3). During February and March of 2011, symptoms of spindle-shaped, straw-colored, irregular lesions with occasional green islands were observed on onion, garlic, and Egyptian leek cultivated on large and small farms in Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiut governorates in Egypt. The presence of IYSV was confirmed by specific double antibody sandwich (DAS)-ELISA Flash Kits (Agdia Inc., Elkhart, IN) (2). A survey was carried out by collecting 100 plant samples (10 asymptomatic and 90 symptomatic) of each plant species from fields in the governorates of Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiute and testing the plants using DAS-ELISA. For onion and garlic, 45% of the symptomatic samples and 0% of the asymptomatic plants tested positive. For leek, 34% of the symptomatic samples tested positive and 0% of the asymptomatic samples. ELISA-positive samples were tested using a reverse transcription (RT)-PCR assay with primers specific to the S RNA of IYSV (forward primer 5'-TAAAACAAACATTCAAACAA-3' and reverse primer 5'-CTCTTAAACACATTTAACAAGCAC-3') (2). Amplicons of approximately 1,100 bp were obtained from all symptomatic samples that were ELISA positive, but none of the asymptomatic plants nor the sterile water control sample produced PCR amplicons. The amplicons were cloned (at least three clones per plant species) using the TOPO TA Cloning Kit (Invitrogen, Grand Island, NY), and sequenced. The Egyptian onion IYSV isolate (GenBank No. JN541273) had the greatest nucleotide sequence identity (86%) with the corresponding S RNA region of IYSV isolates from India (GenBank Nos. EU310290, EU310284, and EU310276). The Egyptian garlic IYSV isolate (GenBank No. JN541275) showed the strongest identity (93%) with that of a Sri Lankan IYSV isolate (GenBank No. GU901211). The Egyptian leek IYSV isolate (GenBank No. JN541274) exhibited 91% sequence identity with that of the same Sri Lankan isolate (No. GU901211). To our knowledge, this is the first report of IYSV infecting garlic and Egyptian leek in Egypt. IYSV infection of onion was reported previously from the agricultural farm of the Faculty of Agriculture, Cairo University, Giza (4), but to our knowledge, this is the first report of natural infection by the virus in commercial onion production in Egypt. Further surveys and monitoring of IYSV incidence and distribution in the entire Egyptian governorate are under investigation. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) A. Manal et al. Egypt. J. Virol. 3:49, 2006.

Non-vascularized free toe phalanx transfers in congenital hand deformities--the Great Ormond Street experience.

Many options of varying complexity are available for the management of congenital short digits resulting from aphalangia in symbrachydactyly and constriction ring syndrome. We have used non-vascularized free toe phalanx transfers for these children when a vascularized toe transfer has been contraindicated. We describe our technique and experience with 22 children who underwent a total of 64 transfers of the proximal (35) or middle (29) toe phalanges (average 3 per child). The mean age at initial surgery was 15 months, and the mean follow-up was 5 years. Duration of time until epiphyseal closure could not be determined accurately, but total digital elongation averaged 6mm. Complications of this technique include joint instability, premature epiphyseal closure and, in one patient, infection and graft loss. Donor site deformity was determined according to measured growth deficit and toe function. This technique is a simple option for digital elongation and, if performed in the appropriate age group in short fingered and monodactylous subtypes of symbrachydactyly, has the potential to allow growth and function with minimal donor site deficit.

Characterisation of the connexin32 promoter and changes in response element complexes in rat liver and hepatocytes during culture associated with oxidative stress.

Hepatic gap junctional intercellular communication (GJIC), mediated principally by connexin 32, provides a mechanism for regulating multicellular activities between neighbouring cells. The control of Cx32 gene expression at the transcriptional level has been investigated in rat liver tissue and in primary rat hepatocytes during culture. Several response elements have been identified and characterised using the electrophoretic mobility shift assay. Nuclear protein extract prepared from rat primary hepatocytes cultured for 2 h gave a larger number of DNA-protein complexes than observed with extracts from liver in vivo, including complexes containing Sp1. In contrast, nuclear extracts prepared from primary rat hepatocytes cultured for 96 h, and subject to oxidative stress, gave altered DNA-protein complexes when compared to those from hepatocytes cultured for 2 h. These results indicate that culture conditions, known to cause a loss of connexin expression, can modulate the transcription of Cx32 in hepatocytes by affecting the regulatory trans/cis-interactions of redox-sensitive zinc finger proteins within the promoter.

Evaluation of the BTA tests for the detection of bilharzial related bladder cancer: the Cairo experience.

OBJECTIVE: To evaluate the clinical performance of the BTA stat test and the BTA TRAK assay in the diagnosis of bilharzia-related bladder cancer and to calculate a new 'Egyptian' cut-off value for the BTA TRAK (quantitative) assay. METHODS: Urine samples of 149 individuals were tested for the presence of the human complement factor H-related protein, the antigen detected by the BTA stat and BTA TRAK tests. The group consisted of 53 healthy volunteers, 20 patients with active bilharziasis, 11 patients with other urologic disorders including prostate cancer, and 65 patients with histologically proven bladder cancer. All samples were obtained prior to surgery or therapy. RESULTS: The BTA stat test was positive in 64 of 65 samples from patients with bladder cancer, for an overall sensitivity of 99%. With a BTA TRAK assay cut-off of 60 U/ml (set at 97% specificity in the healthy population), the sensitivity of the TRAK assay was 94%. There was no statistically significant difference between the sensitivities of the two BTA tests in patients diagnosed with squamous cell carcinoma and those with transitional cell carcinoma. The overall specificity of the BTA stat test was 67% ranging from 15% in patients with bilharziasis to 94% in healthy volunteers. The overall specificity of the TRAK assay was 66%, again with negative results in 15% of the patients with bilharziasis. CONCLUSIONS: The BTA stat test and TRAK tests are extremely sensitive in the detection of bladder cancer in the Egyptian population. Positive results (85%) are also observed in patients with active bilharziasis, which often leads to bladder cancer. Longitudinal follow-up of these positive cases is needed to determine whether these positive results are false or predictive of bladder cancer.

The development of sinoatrial dysfunction in pacemaker patients with isolated atrioventricular block.

The purpose of this paper is the assessment of sinus node competence over time in patients with isolated atrioventricular block (AV block). Patients implanted with AV synchronous pacemakers for isolated AV block between December 1993 and June 1995 were prospectively evaluated at predischarge, 6 weeks, and subsequent 6 months follow-up with respect to atrial rate monitors/24-hour Holter and modified exercise test. Patients unable to maintain AV synchronous pacing or complete a modified exercise test were excluded. Sinus node competency is interpreted as: (1) absence of atrial brady- or tachyarrhythmia, (2) ability to achieve a minimum heart rate of 100 beats/min with modified exercise test or during daily activities. There were 58 patients (22 women), mean age 71.0 +/- 13.8 with an average follow-up of 30.4 months (11-40). Three patients did not complete a modified exercise test, 4 patients were lost to follow-up, and 2 patients were unable to maintain AV synchronous pacing. Of the remaining 49 patients, 3 developed chronic or paroxysmal atrial fibrillation. No patient developed significant bradyarrhythmias. All patients achieved a heart rate of > or = 100 beats/min modified exercise test. In our group of patients with isolated AV block within a moderate follow-up period, development of sinoatrial dysfunction was rare (6%). A longer follow-up is required to delineate the natural history of sinoatrial dysfunction in patients with isolated AV block.

Pituitary apoplexy after leuprolide administration for carcinoma of the prostate.

The syndrome of pituitary apoplexy has been reported to occur after the administration of several different medications. We report a case in which pituitary apoplexy developed shortly after the administration of leuprolide in a patient with prostate cancer. Leuprolide is a potent gonadotrophin releasing hormone (GnRH) analogue used to suppress leuteotrophic hormone (LH) and testosterone levels in patients with metastatic prostate cancer. LH and testosterone levels actually rise in the first week after its administration before becoming suppressed. We suspect that this acute stimulating effect of leuprolide is linked to the acute onset of pituitary apoplexy in a patient with a possible gonadotrophoma.

The spinal fluid of acute hepatitis B patients--an occupational hazard?

Spinal fluid and serum samples from 19 patients, with acute viral hepatitis, type B, already known to have HBs antigenemia, were tested for the detection of HBsAg by two techniques; Hemagglutination Inhibition (H.I.) and Enzyme Linked Immuno Sorbent assay (ELISA). HBsAg was detected in all serum samples by both techniques while all spinal fluid samples were free of that antigen except one sample that was found to contain HBsAg by the ELISA technique only. This sample was also proved to contain occult blood as shown by the positive benzidine reaction.