Pluripotent stem cells in neuropsychiatric disorders.

Neuropsychiatric disorders place an enormous medical burden on patients across all social and economic ranks. The current understanding of the molecular and cellular causes of neuropsychiatric disease remains limited, which leads to a lack of targeted therapies. Human-induced pluripotent stem cell (iPSC) technology offers a novel platform for modeling the genetic contribution to mental disorders and yields access to patient-specific cells for drug discovery and personalized medicine. Here, we review recent progress in using iPSC technology to model and potentially treat neuropsychiatric disorders by focusing on the most prevalent conditions in psychiatry, including depression, anxiety disorders, bipolar disorder and schizophrenia.

Genotypic and pathogenic characterization of genotype VII Newcastle disease viruses isolated from commercial farms in Egypt and evaluation of heterologous antibody responses.

Newcastle disease viruses (NDV) represent a major threat to poultry production worldwide. Recently in Egypt NDV circulated extensively, even in vaccinated farms. In the present study samples were collected from sixteen vaccinated broiler farms in animals exhibiting the typical gross lesions of NDV. Virus isolation and pathogenicity studies for positive samples were carried out in accordance to reference procedures and phylogenetic analysis was carried out based on partial sequences of the Fusion gene. Furthermore, in vivo investigation of the ability of heterologous antibody, induced by commercially available lentogenic strain-based vaccines, to efficiently reduce viral shedding was examined. Results revealed that all the sixteen farms were positive for the presence of NDV. Out of these fifteen were confirmed to due to velogenic viruses, based on a main death time (MDT) ≤ 48 hours and partial sequencing of the F gene that showed the presence of a polybasic amino acid motif. However, three patterns in the cleavage site of these velogenic viruses were identified in the present study. Phylogenetic analysis revealed that all fifteen isolates were clustered with class II genotype VIIb while the remaining isolate (B81) was class II genotype II. Results of the in vivo study revealed that adequate heterologous antibody levels, induced by the proposed vaccination program, sufficiently protected birds from morbidity and mortality. However, virus shedding was quantitatively affected in relation to the time of challenge after vaccination. Altogether, with an absence of vaccines able to induce homologous antibody to the presently circulating viruses, higher antibody levels, which depend on efficient and timely implementation of the vaccination program, are considered as highly important in relation to the reduction of virus shedding.

Effect of preservative-free and preserved prostaglandin analogues on the histology of cornea of adult male guinea pigs following repeated exposure.

BACKGROUND: Glaucoma is a group of eye diseases that can cause vision loss. Prostaglandin analogues (PGAs) are known to be first-line treatment for patients with glaucoma. Latanoprost is a good, efficient and well-tolerated PGA that is currently available as latanoprost with benzalkonium chloride (BAC) (Xalatan) and preservative-free (PF) prostaglandin analogue latanoprost (Monopost). Lately, using PF anti-glaucoma agents has been considered an essential procedure for enhancing glaucoma care. This study aimed to analyse the histological changes within the corneal tissue with the use of currently available preserved prostaglandins-derived eye drops and PF prostaglandin analogue. MATERIALS AND METHODS: In this study, 40 male guinea pigs were divided into four equal groups. Control group, Latanoprost with 0.02% BAC-treated group, Recovery group and PF latanoprost-treated group. After 2 months, the corneal tissues of guinea pigs were prepared for light and electron microscopic studies; morphometric and statistical studies were performed. RESULTS: Our results indicated that guinea pigs treated with latanoprost with BAC exhibited ocular surface changes: there was epithelial thinning with desquamation, the stroma showed irregularly arranged collagen fibres and small keratocytes. Morphometrically, there was a marked decrease in the thickness of epithelium and number of keratocytes. Negative periodic acid Schiff (PAS) reaction was observed in some parts of the epithelial basement membrane. The epithelium gave a strong positive immunoreactivity for Bcl-2-associated X protein (BAX). Guinea pigs left to recover exhibited improvement, while treatment of animals with PF latanoprost resulted in nearly normal corneal structure. CONCLUSIONS: In conclusion, PF prostaglandin anti-glaucoma medication seems to be better and have protective effect on cornea of male guinea pigs than prostaglandins with BAC preservative.

Development of hydrogel based on Carboxymethyl cellulose/poly(4-vinylpyridine) for controlled releasing of fertilizers.

A novel Carboxymethyl cellulose (CMC) and poly (4-vinylpyridine) (P4VP) hydrogel system is synthesized with different ratios, in the presence of cross-linker N, N,- methylene bis-acrylamide (MBA). The hydrogel is characterized via FTIR spectroscopy, thermal gravimetric analysis (TGA), X-ray diffraction (XRD), and scanning electron microscope (SEM). The FTIR results showed a strong interaction between both CMC, P4VP and the loaded fertilizer. The water uptake of the hydrogel was evaluated by swelling tests under variations in pH, biodegradability was investigated in soil to simulate real-world conditions. To determine the best release behavior of urea and calcium nitrate from the hydrogel, fertilizers were loaded with different ratios onto the hydrogel during its formation. Fertilizers release was followed by Atomic absorption spectroscopy to study the release of calcium nitrate and urea. Release kinetic parameters were obtained based on different mathematical models as Zero order, First order, Korsmeyer-Peppas and Higuchi models. The suitable proportionality between the mathematical models used and the fertilizers release was determined based on the correlation coefficients (R2). According to Zero order model urea release showed independent concentration. Based on Korsmeyer-Pappas and Higuchi models with high n value and R2 equals to 0.97. Compared to urea, Ca2+, Zero order and Higuchi have been ignored due to their poor correlation coefficients values as proportion with Ca2+ fertilizer release.

Serum Fetuin A Levels: Are They a Reliable Marker for Hepatic Steatosis and Regional Adiposity in Renal Transplant Recipients?

BACKGROUND: Fetuin A is a protein expressed in the liver and it is an important inhibitor of ectopic calcification. High levels of fetuin A correlate with insulin resistance, hepatic steatosis, and regional adiposity in the general population. The association between hepatic steatosis and fetuin A level in renal transplant recipients (RTRs) remains unclear. AIM: The aim of this study was to explore the relationships between fetuin A, hepatic steatosis, and regional adiposity in RTRs. METHODS: Data from 44 patients with normal renal functions were included, all subjected to history taking for clinical data, assessment of central obesity and regional adiposity, assessment of hepatic steatosis using abdominal ultrasound (US), and measurements of serum fetuin A concentration using enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: Our study included 20 females (45.4%) and 24 males (54.6%) with mean age of 41.26 ± 11.2 years. Twenty-four subjects had hepatic steatosis. Fetuin A level in RTRs with hepatic steatosis with a mean of 1642.92 ± 358.91 is significantly higher (P < .001) than those without hepatic steatosis with a mean of 711.74 ± 57.85. Serum fetuin A level was positively correlated with regional adiposity (P = .021) and hepatic steatosis grade (P = .017). Fetuin A level increased with increased duration after renal transplantation (P < .001). The best cutoff value for detecting entrance into phase 3 or 4 steatosis is fetuin of 1862 with sensitivity of 88.9% and specificity of 87.7%. CONCLUSIONS: Fetuin A is positively correlated with hepatic steatosis and regional adiposity in RTRs. Fetuin increases with increased duration after renal transplantation. Accordingly it may be used as a marker for hepatic steatosis and regional adiposity in these patients.

IL-17 Producing Cells and RORγt mRNA Transcriptional Factor in Cirrhotic and HCC Egyptian Patients.

Retinoic acid-related orphan receptor gT (RORγT) is the orphan nuclear receptor that regulates the development of Th17 cells and the expression of IL-17. The differentiation of Th17 cells is associated with the upregulation of RORγT mRNA, and the mechanisms regulating that process in human cells are not well understood. RORγt as transcription factor was selectively expressed in Th17 cells and is regulated by STAT3. The relationship between Th17 cells and tumor immunopathology has been controversial. Aim of the study is to evaluate Th17 cells and RORγt transcriptional factor in cirrhotic, early and advanced HCC patients. Ninety patients were studied (30 cirrhotic, 30 early stage and 30 advanced stage HCC patients). They were recruited from the National Liver Institute, Menoufia University, and subjected to full clinical examination, investigations to detect liver cirrhosis, portal vein thrombosis and tumor staging. Peripheral blood mononuclear cells (PBMCs) were stimulated with cytosim, ionomycin and monensin, and surface (CD4-PE)as well as intracellular staining for Th17 (IL-17 FITC) was performed. Analysis of cells was carried out using FACS Calibur. RORγt mRNA expression in PBMCs was measured by real time RT- PCR. Cirrhotic patients showed increased Th17% cells, without significant change in RORγt mRNA as compared to early stage HCC. Advanced stage HCC patients showed significant increase of Th17 cells% and RORγt mRNA compared to studied patients group. Positive correlation of Th17 and RORγtmRNA was found with aminotransferases and bilirubin levels while, negative with serum albumin in advanced stage patients group (P < 0.001). Both markers were significantly increased with tumor size; RORγt mRNA increased with multiple tumor foci. In conclusion, Th17 cells and RORγt may be useful prognostic markers for advanced liver cirrhosis and HCC.

Effect of hydrocortisone on immune system of the lizard, Chalcides ocellatus. III. Effect on cellular and humoral immune responses.

Administration of a single injection of 1.0 mg/g body weight hydrocortisone acetate (HC) to adult lizards, Chalcides ocellatus in summer elicited a high and lasting rise in serum corticosterone and cortisol, peak levels being, however, in the physiologic ranges of 10 micrograms % (2 X 10(-7) M) and 40 micrograms % (1 X 10(-6) M), respectively. Elevation of serum corticosteroid (CS) concentrations by exogenous HC impaired the proliferative capacity of spleen cells in mixed leukocyte reaction (MLR) and significantly delayed skin allograft rejection. In vivo HC also abrogated the primary in vivo and in vitro immune responses of lizards to rat erythrocytes (RRBC). Reactivity to allogeneic cells in MLR, and antibody production against RRBC were, however, recovered when serum CS levels resumed normal, basal values i.e. at three weeks post-HC injection. These data indicate that rise in circulating CS induces severe immunosuppression in lizards.

Concanavalin A responsiveness and interleukin 2 production in the snake Spalerosophis diadema.

Thymocytes and splenocytes (SC) of adult snakes, Spalerosophis diadema, responded to concanavalin A (Con A) in vitro by strong proliferation during the spring and autumn seasons. Con A-mediated mitogenesis was, however, abrogated in summer and winter. Conditioned medium (CM) collected from snake SC cultures stimulated with Con A in spring or autumn could enhance the Con A summer and winter responses and support the proliferation of splenic lymphoblasts. Gel filtration of native CM on Sephadex G-100 revealed the presence of two biologically active peaks of molecular weight 39-42 and 15 KD. However, only one peak of activity corresponding to molecular weight (m.w.) of 14-15 KD was observed when CM was subjected to analysis by sodium dodecyl sulphate gel electrophoresis (SDS-PAGE). The active molecular forms exhibited isoelectric points of 5.5-5.8 and 6.4-6.6. The findings suggest that Con A activation of snake lymphocytes in optimal seasonal conditions is associated with the secretion of a lymphokine analogous to the interleukin 2 (IL 2) of endothermic vertebrates.

Distance from treatment facility and risk of death from cardiovascular and infectious causes in renal transplant patients.

We investigated whether patients receiving RTx who live farther from their attending nephrologist are more likely to die than those who live closer. A random sample of 167 patients who undergone RTx between 1996 and 2004 was examined. We calculated the distance between each patient's residence and the practice location of their attending nephrologist. We used Cox proportional hazards models to examine the adjusted relation between distance and clinical outcomes (death from all causes, rejection episodes, infectious causes, and cardiovascular complications) over a follow-up period of upto 6 years. During the follow-up period (median: 3.3, range: 1.0-6.5 years), 22% of patients died. Compared with patients who lived within 50 km of their nephrologist, the adjusted hazard ratio of death was 1.04 among those who lived 50.1-150 km away, 1.16 for those who lived 150.1-300 km away, and 1.19 for those who lived more than 300 km (P for trend <0.001). The risk of death from infectious causes increased with greater distance from the attending nephrologist (P for trend <0.001). The risk of developing acute rejection episodes did not increase with distance from the attending nephrologist (P for trend = 0.2). The risk of death from cardiovascular causes increased with distance from the attending nephrologist (P for trend <0.05). Compared with patients who lived within 50 km of their nephrologist, the adjusted hazard ratio of death among those who lived >300 km away was 1.75 for infectious causes and 1.39 for cardiovascular causes. We conclude that mortality and morbidity associated with RTx was greater among patients who lived farther from their attending nephrologist, as compared with those who lived closer.

Further evidence of antigenic drift and protective efficacy afforded by a recombinant HVT-H5 vaccine against challenge with two antigenically divergent Egyptian clade 2.2.1 HPAI H5N1 strains.

In this study, we have compared the protection afforded by a recombinant turkey herpesvirus vaccine expressing the H5 gene from a clade 2.2 H5N1 strain (rHVT-H5) and a Mexican-origin H5N2 inactivated vaccine, alone or in combination, against two antigenically divergent H5N1 Egyptian strains isolated in 2007 and 2008. Our results confirm the existence of a major antigenic drift among the Egyptian H5N1 strains such that, although protection against the "classical" 2007 HPAI H5N1 Egyptian strain could be obtained with both types of vaccines, only vaccination with the rHVT-H5 vaccine protected against challenge with the "variant" 2008 HPAI H5N1 Egyptian strain.

Antiplasmodial and antitrypanosomal activity of plants from the Kingdom of Saudi Arabia.

The antiplasmodial and antitrypanosomal activity of the methanol extracts of 42 plants collected from the Kingdom of Saudi Arabia and some fractions obtained thereof were evaluated. The antiplasmodial activity was tested in vitro against chloroquine-resistant strain (K1) and sensitive strain (FCR3), and the antitrypanosomal activity was tested in vitro against Trypanosoma brucei brucei GUTat 3.1 strain. For host cells, the cytotoxicity of the active extracts was also evaluated against the MRC5 human cell line. Only extracts of three samples demonstrated good antiplasmodial activity (IC(50) < 12.5 and > 1.56 microg/ml, score 2), the methanol extracts of Lycium shawii, Heliotropium zeylanicum and the petroleum ether-soluble fraction of the methanol extract of Caralluma tuberculata, while extracts of the remaining 42 plants were inactive (IC(50) > 12.5 microg/ml, score 1). As for the antitrypanosomal activity, the methanol extract of Solanum schimperianum demonstrated the highest activity (IC(50) 0.061 microg/ml), followed by the petroleum ether-soluble fraction of the methanol extract of C. tuberculata (IC(50) 0.5 microg/ml). The chloroform-soluble fraction of the methanol extract of C. tuberculata was moderately active (IC(50) 3.5 microg/ml), with low cytotoxicity (IC(50) 62.6 microg/ml) and moderate selectivity index (SI 17.9). The methanolic extracts of 34 plants showed good activity with score 2 (IC(50) < 12.5 and > 1.56 microg/ml), while the extracts of seven plants were inactive (IC(50) > 12.5 microg/ml, score 1).

Phase-contrast study of the efficacy of Lampit (BAY 2502, Nifurtimox) on experimental infection with Trypanosoma rhodesiense.

Lampit (BAY 2502, Nifurtimox) was tested in experimental Wistar strain white rats with acute infection induced by Trypanosoma rhodesiense strain (180 TS1A1), which normally contains cytoplasmic granules of both Type (I) & (II). Lampit injected subcutaneously in low doses of 25, 50, 100, and 250 mg/kg b-w as one-time application, gave no effect. When repeated higher doses (500 mg/kg b-w) was given a reasonable effect was achieved and formation of additional granules in T. rhodesiense was noticed. In experimental animals, some what higher initial doses appear to be particularly advantageous.

In vitro study on generation time (G) of Trichomonas vaginalis.

31 strains of T. vaginalis were isolated from female patients attending venereal diseases clinic, Middlesex Hospital, London. Cloned organisms were studied in vitro for generation time (G). According to generation time T. vaginalis strains were divided into three different groups: slow, medium and fast grow.

Comparative study of the effect of atropine, P2AM, and Buscopan. Total belladonna and hyoscyamus alkaloids on parathion poisoning.

Among the available antidotes used against parathion poisoning, atropine and Buscopan are tolerated in relatively high doses by rats. Hyoscine (in very small doses), T. Bel. Alk. and T. Hyosc. Alk. (better results in small doses, also for dogs), P2AM (of superior antimuscarine action) are redommended with each of the preceding drugs than either alone or in combination with Buscopan.

Isoenzyme characterisation of Trichomonas vaginalis.

Clones of 32 strains of Trichomonas vaginalis isolated from patients attending a venereal diseases clinic were compared among themselves and with authentic Pentatrichomonas hominis on the basis of their isoenzyme patterns for eight enzymes by thin-layer starch-gel electrophoresis. The enzymes examined were: glucose phosphate isomerase (GPI); phosphoglucomutase (PGM); malic enzyme (NADP+) (ME); hexokinase (HK); malate dehydrogenase (NAD+) (MDH); glucose-6-phosphate dehydrogenase (G6PD); aldolase (ALD); and lactate dehydrogenase (LDH). From the isoenzyme patterns of four enzymes (LDH, MDH, HK, and GPI) the strains of T vaginalis could be divided clearly into five groups. PGM showed differences in only one strain, while two other enzyme patterns (ME and ALD) were the same for all the strains of T vaginalis tested. All isolates were clearly distinguishable from P hominis. Although G6PD patterns were not sharp some differences were evident among T vaginalis strains.

Thymus histogenesis in the lizard, Chalcides ocellatus.

Different stages of thymus morphogenesis and thymocyte differentiation have been studied at the ultrastructural level in the lizard, Chalcides ocellatus. On stage 36 of embryonic development, the thymus primordium was composed principally of undifferentiated epithelial cells and some lymphoid stem-cells. From stage 37 to 38, the lymphoid stem-cells differentiate into lymphoblasts and then transform into typical lymphocytes. A clasmotosis phenomenon seems to be involved in this transformation. In the developing cortical regions, lymphoblasts accumulated rapidly, stretching the epithelial cells which become stellate in shape. From stage 39 to 40, a phase of intense proliferation occurs and numerous lymphocytes die in the thymic tissue and are phagocytosed by macrophages. On stage 41, the presence of interdigitating cells in the medullary area completed cortico-medullary differentiation. On neonatal and juvenile lizards, small cortical thymocytes differentiated and the thymus possessed all characteristic of an adult thymus. Thus, at birth, the histogenesis of the lizard thymus was achieved and the only further modification consisted in a gain of weight.

Development of Wuchereria bancrofti in Culex pipiens L. (Diptera: Culicidae) exposed in the larval instar to sublethal dosages of insecticides and one insect growth regulator and their influence on reproduction of filaria-infected mosquitoes.

The effects of exposure of Culex pipiens larvae to sublethel concentrations of larvicides on uptake, development of Wuchereria bancrofti, survival rate and reproduction of filaria-infected mosquitoes were investigated. Fourth instar larvae of Cx. pipiens were exposed to LC40 of the surfactant Triton X-100, the insect growth regulator DPX alone or combined with LC10 of the surfactant and permethrin alone or combined with LC10 of the surfactant. Adults that survived insecticide treatments and controls were infected by allowing them to feed on microfilaremic volunteers. Significant reduction in the uptake of microfilaria was observed in groups treated with Triton X-100 alone or combined either with permethrin or DPX when compared to control. The overall infection and infective rates were significantly reduced in mosquitoes treated with Triton X-100 either alone or combined with permethrin. Treatment with Triton X-100 and DPX prolonged the extrinsic incubation period (EIP) and retarded the development of filarial larvae, while permethrin either alone or combined with Triton X-100 and DPX combined with Triton X-100 shortened the EIP. All larvicides reduced the number of infective larvae (L3)/mosquito and induced deformities among he different parasite stages, especially in mosquitoes treated with combination of permethrin and Triton X-100 or mixture of DPX and Triton X-100 where 36% and 54.9% respectively of L3S were deformed. In treated mosquitoes, a low percentage of L3S was detected in the head and proboscis region while the majority was trapped in the thoracic region. The survival rates of mosquitoes were reduced in cases treated with permethrin, DPX and Triton X-100 while treatment with mixture of DPX and Triton X-100 induced higher rate of mortalities when compared to control. Egg production of filaria- infected Cx. pipiens was significantly reduced in mosquitoes treated with DPX and Triton X-100. It was observed that the addition of Triton X-100 to DPX or to permethrin significantly reduced egg production. The results suggest that sublethal concentrations of larvicides especially Triton X-100 applied to 4th instar larvae of Cx. pipiens could effectively interfere with the development of W. bancrofti in Cx. pipiens and reduced the survival rate and fecundity of the vector.

Studies on heterologous resistance between schistosomiasis mansoni and fascioliasis.

S. mansoni and F. gigantica GSTs were purified from adult worm homogenates by affinity chromatography. Assessment of both SmGST and FgGST as candidate vaccines was done by comparing vaccinated mice groups challenged with S. mansoni cercariae with control ones in terms of worm load, tissue egg number viability and maturity, and the survival rate of the animals. SmGST 5 ug vaccination generally appeared to confer best protection against homologous challenge followed by FgGST 5 ug while vaccination with both antigens using low doses (2 ug) appeared to have no specific role in decreasing worm load but had significant effects on egg production.