Simultaneous estimation of troxerutin and calcium dobesilate in presence of the carcinogenic hydroquinone using green spectrofluorimetric method.

In the present study, we conducted two facile and highly sensitive spectrofluorimetric approaches in order to quantify the vasoprotective agents; troxerutin (TROX) and calcium dobesilate (DOB) in the presence of hydroquinone (HQ) (as a highly toxic impurity and potential degradation product of DOB) in commercial formulations and human plasma. The first approach relies simply on using ethanol as an eco-friendly solvent for the estimation of DOB at 345 nm after being excited at 305 nm. The linearity was carefully investigated between DOB concentration and the relative fluorescence intensity in the range of 0.05-0.8 µg ml-1. Due to the high method simplicity and sensitivity, applying the first approach to quality control analysis and spiked human plasma samples with mean % recoveries 100.74 ± 3.71 adds another merit. The second approach involved rapid conventional fluorimetric estimation of ethanolic TROX solution in TROX/DOB combined dosage forms at 455/350 nm (emission/excitation) with a linear calibration chart covering the range of 0.1-1.2 µg ml-1. Moreover, the second approach involved a comprehensive study in a trial to solve the problem of superposition of DOB and HQ graph adopting the first derivative synchronous fluorimetric mechanism in ethanol at Δλ = 60 nm. Therefore, DOB was measured at 286 and 323 nm, while HQ could be quantitated at 301 nm. The Beer-Lambert Law has complied over the ranges of 0.1-1.0 and 0.02-0.4 µg ml-1 for DOB and HQ, respectively. Guidelines adopted by the International Council of Harmonization (ICH) were used to validate the target approaches. The developed methods are more convenient for routine quality control laboratory instead of the time-consuming and sophisticated reported techniques. Moreover, different aspects of evaluating the greenness of the proposed approaches were conducted to have a complete image of their environmental impact.

Hydrogen sulfide ameliorates cardiovascular dysfunction induced by cecal ligation and puncture in rats.

Hydrogen sulfide (H2S) is an endogenously produced gaseous messenger that participates in regulation of cardiovascular functions. This study evaluates the possible protective effect of H2S in cardiovascular dysfunction induced by cecal ligation and puncture (CLP) in rats. After 24 h of induction of CLP, heart rate (HR), mortality, cardiac and inflammation biomarkers (creatine kinase-MB (CK-MB) isozyme, cardiac troponin I (cTnI), C-reactive protein (CRP), and lactate dehydrogenase (LDH)), in vitro vascular reactivity, histopathological examination, and oxidative biomarkers (malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD)) were determined. CLP induced elevations in HR, mortality, serum CK-MB, cTnI, CRP, and LDH, in addition to impaired aortic contraction to potassium chloride and phenylephrine and relaxation to acetylcholine without affecting sodium nitroprusside responses. Moreover, CLP increased cardiac and aortic MDA and decreased SOD, without affecting GSH and caused a marked subserosal and interstitial inflammation in endocardium. Sodium hydrosulfide, but not the irreversible inhibitor of H2S synthesis dl-propargyl glycine, protected against CLP-induced changes in HR, mortality, cardiac and inflammatory biomarkers, oxidative stress, and myocardium histopathological changes without affecting vascular dysfunction. Our results confirm that H2S can attenuate CLP-induced cardiac, but not vascular, dysfunction possibly through its anti-inflammatory and antioxidant effects.

Spontaneous clearance of chronic hepatitis C infection is associated with an internal ribosomal entry site IIId stem loop structure variant.

AIM: To investigate if any mutations in hepatitis C virus (HCV) internal ribosome entry site (IRES) can inhibit the translation of viral polyprotein. MATERIALS AND METHODS: A 26-year-old male patient infected with HCV 10 years ago was followed up. After 9 years of chronic infection. The patient had managed to resolve the infection for a period of 9 months, after which the patient experienced a viral recurrence characterized by high viral load and diverse HCV quasispecies. The IRES structures of the viral strains that disappeared were comparable with those that are currently active using structural mutational analysis. RESULTS: A novo mutational position 254 combined with a rarely observed mutation at position 253 in the stem of the IIId subdomain were observed and the new conformation had an octa-apical loop (AGUGUUGG) and a shift in the 3 ` GU from the loop to the stem. CONCLUSIONS: These mutations were found to be highly deleterious, and they affected the direct binding of the IIId loop to the 40S ribosomal subunit with a subsequent inhibition of translation of viral polyprotein and clearance of the virus.

Air quality in Ain Shams University Surgery Hospital.

Through air sampling, it was possible to evaluate microbial contamination in environments at high risk of infection, and to check the efficiency of ventilation system and the medical team's hygiene procedures. This study measured the concentration of particulate matter (PM) 2.5 or less microns and microbiological organisms in operating rooms (OR), intensive care units (ICU) and emergency rooms (ER) in Ain Shams University Surgery Hospital, and to assess ventilation characteristics in operating rooms in the hospital. The passive air sampling was done from ICUs, ORs, and ERs in Ain Shams University Surgery Hospital. Also for each operating room, an observational checklist was done to record other factors that may affect air quality in the room. The evaluated air quality indices were: suspended (PM) 2.5 micrometer or less, culture media and microbial identification of bacteria and fungi, and temperature and relative humidity. The results showed that the highest mean found for bacterial (105.70±30.49) and fungi concentration (7.50±5.30) was in ER. The three settings did not differ statistically as regard levels of PM 2.5, temperature, and relative humidity. A positive correlation exits between bacteria and fungi concentration on one hand and relative humidity on the other. Diphteroid, CONS, MRSA, S. aureus, and Anthracoid were the most frequent isolated bacterial types, while Penicillium and Asperigillus fumigatus were the most frequent isolated fungi. In operating rooms, the percent of unmasked persons present and the temperature positively influence the bacterial count, while ventilation condition is negatively influencing fungi count, and the number of persons present in the operating room positively affects the PM level.

Fluorometric determination of bopindolol and celiprolol in pharmaceutical preparations and biological fluids.

Two sensitive fluorometric methods were developed for the determination of both bopindolol malonate (BOP) and celiprolol HCl (CLP) based on measuring their native fluorescence in methanol and acetonitrile, respectively. For BOP, the fluorescence was measured at 316 nm after excitation at 278 nm. The proposed method was successfully applied to the assay of commercial tablets as well as content uniformity testing. For CLP, the fluorescence was enhanced by the addition of carboxymethylcellulose solution and measured at 455 nm after excitation at 339 nm. The method was successfully applied to the analysis of CLP in tablets and biological fluids. In both methods, interference likely to be introduced from co-formulated, co-administered, or chemically related drugs was studied. The results were statistically compared with those obtained by reference methods and were found to be in good agreement.

pH-sensitive sodium alginate hydrogels for riboflavin controlled release.

Sodium alginate (SA) grafted with polyglycidyl methacrylate hydrogels (PGMA-g-SA) was prepared as pH sensitive drug delivery matrices for riboflavin (RF). The hydrogel copolymer matrices were compared with calcium alginate (CA) beads for swelling, degradation, entrapment efficiency and in vitro release of RF. The structure, surface morphology of the CA beads and the prepared hydrogels as well as the chemical stability of the encapsulated drug were characterized by FT-IR and SEM, respectively. The results demonstrate that the optimal formulation was achieved with PGMA-g-SA proportion of (0.75 mol/1 g) and loaded RF 0.03 g. It has been observed that the in vitro release study of RF from this formulation was superior to the other ones and was able to maintain the release for ∼3 and 4 days for the simulated intestinal fluid (SIF) and simulated gastric fluid (SGF), respectively. In general, it has been shown that, GMA grafted onto SA enhanced drug entrapment efficiency, decreased swelling and degradation behaviors of the carrier. In addition, it slowed and controlled the release of RF from the PGMA-g-SA hydrogel compared with pure SA beads crosslinked with Ca2+ ions alone, which thereby provides a facile and effective method to improve the drug delivery systems.

Desensitization of the soluble guanylyl cyclase/cGMP pathway by lipopolysaccharide in rat isolated pulmonary artery but not aorta.

BACKGROUND AND PURPOSE: To investigate the function of soluble guanylyl cyclase (sGC)/3',5'-cyclic guanosine monophosphate (cGMP) pathway in lipopolysaccharide (LPS)-induced changes in vascular reactivity of rat isolated pulmonary artery and aorta. EXPERIMENTAL APPROACH: Nitric oxide (NO) production, contraction responses to endothelin-1 (ET-1), relaxation responses to sodium nitroprusside (SNP), 8-pCPT-cGMP, BAY412272 and T-0156, SNP-induced cGMP production and expression of sGC(alpha1), sGC(beta1) and 3',5'-cyclic nucleotide phosphodiesterase-5 (PDE5) proteins were measured in LPS-treated pulmonary and aortic rings from male Wistar rats. KEY RESULTS: In both vessels, LPS (10 microg mL(-1), 20 h) increased NO production, which was inhibited by the selective inducible NOS (iNOS) inhibitor 1400W (1 microM). In the aorta, LPS decreased ET-1-induced contractility and this decrease was inhibited by the selective sGC inhibitor ODQ (10 microM) but not by removal of endothelium, or inhibitors of cyclooxygenase (indomethacin, 10 microM) or iNOS (1400W, 1 microM). Furthermore, aortic relaxation responses to the direct sGC activator BAY412272 were enhanced. In the pulmonary artery, SNP (1 nM to 30 microM)-induced relaxation and cGMP production, BAY412272-induced relaxation and sGC(beta1) protein expression were decreased, whereas relaxation responses to the PDE5-specific inhibitor T-0156 (0.1-100 nM) were enhanced. Relaxation responses to the phosphodiesterase-resistant cGMP analogue, 8-pCPT-cGMP, and protein expression levels of sGC(alpha1) and PDE5 were not altered in either vessel. CONCLUSION AND IMPLICATIONS: LPS caused a selective hypocontractility of rat aorta to ET-1 mediated mainly through NO-independent sGC activation, whereas in the pulmonary artery, the effect of sGC activation was reduced by a decreased protein expression of sGC(beta1) together with increased PDE5 activity.

Status of diphtheria immunity in the Egyptian population.

In an attempt to evaluate the levels of immunity against diphtheria in the Egyptian population, 709 healthy subjects aged 2 months-105 years and from six regions of Egypt were investigated. An ELISA was used to determine the serum concentration of anti-diphtheria IgG in each subject. Following widely-used criteria for defining levels of protection, 34.1% of the subjects were categorized as susceptible to diphtheria (with < 0.01 IU antitoxin/ml serum), 43.7% were considered to have basic protection (0.01- < 0.1 IU/ml) and only the other 22.1% appeared fully protected (> or = 0.1 IU/ml). The results revealed that most of the subjects aged 2 months-50 years had a basic or fully protective level of IgG against diphtheria, although males were slightly more likely to be unprotected than females (36.2% v. 31.6%) and certain age-groups appeared to be much more likely to be susceptible than others. If outbreaks of diphtheria like those seen in recent years in the former Soviet Union are to be avoided in Egypt, the most susceptible groups of the population need to be given booster immunizations.

The milder phenotype of the dystrophin gene double deletions.

OBJECTIVES: This study aimed to examine the genotypephenotype correlation in Duchenne muscular dystrophy (MD) patients with double deletion (Ddel) mutations in comparison with those having single deletions (Sdel). MATERIALS AND METHODS: The study included 250 Duchenne/Becker MD male patients from whom the 10 Ddel patients were compared with 20 Sdel subjects of same age and disease durations. The patients were subjected to neurological examination including functional disability grading scale (FDGS), molecular analysis of the dystrophin gene and immunohistochemical studies of some muscle biopsies. RESULTS: The mean FDGS value in the Ddel group was lower than that in Sdel patients. The Ddel patients had partial expression of dystrophin in their skeletal muscles, while Sdel cases showed complete absence of the protein. CONCLUSION: Patients with double deletion mutations within the dystrophin gene have a milder phenotype than patients harboring single deletions at either major or minor hot spots of the gene.

Direct enantiomeric resolution of some cardiovascular agents using synthetic polymers imprinted with (-)-S-timolol as chiral stationary phase by thin layer chromatography.

Molecular imprinted polymers (MIPs) of S-timolol were prepared as chiral stationary phases (CSPs) in thin layer chromatography (TLC). The resolution of the enantiomers of some cardiovascular drugs, including propranolol, atenolol, timolol, nadolol, nifedipine and verapamil were investigated on these CSPs. A mobile phase system of either methanol or acetonitrile was used and the effects of acetic acid content of the mobile phases were also investigated. The best resolution was achieved for enantioseparation of propranolol, timolol and atenolol on plates based on MIP of (-)-S-timolol using methacrylic acid as functional monomer (alpha = 1.52, 1.6, 1.59) respectively, using acetonitrile containing 5% acetic acid and (alpha = 1.47, 1.52, 1.5) in methanol containing 1% acetic acid as mobile phases. The results obtained show that TLC based on MIPs could be applied in the direct separation of several beta-adrenergic drugs. As the side chains on beta-blockers are similar, it is possible that this method could also be used for the resolution of other racemates in this family of drugs. Racemic drugs structurally related to print molecule, were completely resolved into two spots with the MIP plates. In general the retention of (+)-R-isomers was greater than that of (-)-S-isomers, indicating lower stereoselectivity of the MIPs to the dextrorotatory isomers. The method offers a rapid, sensitive and reliable method for quality control for these drugs.

In utero exposure to cadmium pollution in Cairo and Giza governorates of Egypt.

Serum cadmium levels at delivery were measured in a consecutive sample of 100 mother-infant pairs in Egypt using venous blood from mothers and umbilical cord blood from neonates. The serum cadmium levels of mothers ranged from 0.4 to 2.2 microg/L (mean 0.73 microg/L) and of infants from 0.2 to 1.5 microg/L (mean 0.66 microg/L). Infant cadmium levels were about 70% of maternal levels in most pairs. Serum cadmium was significantly higher in mothers and babies passively exposed to tobacco smoke. Five-minute Apgar scores were negatively correlated with cord blood cadmium levels. The cadmium levels did not differ between subjects from Cairo and Giza or according to urban, suburban or rural areas. Thus, in utero exposure to cadmium was evident and wider-scale studies on its long-term effects are recommended.

In utero exposure to cadmium pollution in Cairo and Giza governorates of Egypt

Serum cadmium levels at delivery were measured in a consecutive sample of 100 mother-infant pairs in Egypt using venous blood from mothers and umbilical cord blood from neonates. The serum cadmium levels of mothers ranged from 0.4 to 2.2 microg/L [mean 0.73 microg/L] and of infants from 0.2 to 1.5 microg/L [mean 0.66 microg/L]. Infant cadmium levels were about 70% of maternal levels in most pairs. Serum cadmium was significantly higher in mothers and babies passively exposed to tobacco smoke. Five-minute Apgar scores were negatively correlated with cord blood cadmium levels. The cadmium levels did not differ between subjects from Cairo and Giza or according to urban, suburban or rural areas. Thus, in utero exposure to cadmium was evident and wider-scale studies on its long-term effects are recommended

Schistosoma hematobium soluble egg antigens induce proliferation of urothelial and endothelial cells.

Bladder carcinoma accounts for 26% of reported human malignancies in Egypt, and has been strongly associated with urinary schistosomiasis. Nevertheless, the immediate role of schistosomal egg proteins in bladder carcinogenesis is unexplored. We investigated the effects of crude soluble egg antigens (SEA) of Schistosoma hematobium on urothelial cell proliferation. The proliferation of bovine endothelial Endo, human urothelial J82 and smooth muscle SMC cell lines was assessed by low-density growth assays. SEA induced proliferation of both J82 and Endo cells in a dose-dependent fashion, but not SMC. Preboiling or proteinase K treatment of SEA abolished its effect. In addition, SEA enhanced urothelial expression of B-cell translocation protein (BTG1) and human proliferating cell nuclear antigen (PCNA) mRNAs. Given the strong correlation between cell proliferation and carcinogenesis, the findings suggest that crude SEA may play some role in schistosomal bladder carcinogenesis.

Environmental exposure of the pediatric age groups in Cairo City and its suburbs to cadmium pollution.

In a trial to assess the exposure of subjects in the pediatric age group to cadmium (Cd) pollution, serum Cd was estimated by atomic absorption spectrometry in 405 subjects, birth-18 years old, from Cairo City and its suburbs. Serum Cd mean concentrations were: 0.92 microg/l in 32 neonates (birth-4 weeks); 1.33 microg/l in 70 infants (4 weeks-2 years); 1.11 microg/l in 100 children in the preschool period (2-6 years); 1.34 microg/l in 103 primary school children (6-12 years); and 1.24 microg/l in 100 adolescents (12-18 years). In neonates, serum Cd was higher in babies with weights and heights that remained below the 5th percentile for age. Breast-fed infants had a serum Cd geometric mean level (1.25 microg/l) that was not in accordance to that of their mothers' milk (0.52 microg/l, P < 0.001), suggesting alternative routes of exposure. Environmental tobacco-smoke exposure was the most important determinant of Cd status in the school-aged children, the geometric mean being 1.42 microg/l in passive smokers vs. 1.2 microg/l in non-exposed children (P < 0.05). Moreover, adolescents who were active smokers had a significantly higher serum Cd level (1.7 microg/l) as compared to non-smokers (1.2 microg/l). Gender did influence the Cd status in adolescents, being higher among males, probably related to smoking, or to the difference in lifestyle of adolescents according to gender in the community. Alpha-1-microglobulinuria was accompanied by a higher serum Cd concentration in the group of adolescents only, suggesting a subclinical renal effect after several years of cumulative exposure. The residential classification, whether urban or suburban, did not influence the serum Cd status; neither did the present or past history of bronchial asthma. These findings certainly justify further evaluation of the problem of Cd pollution among Cairene individuals, knowing the long-term consequences of exposure to it. Systematic efforts for the proper disposal of Cd wastes and prevention of smoking in public places are recommended.

Differential display analysis of mRNAs in chronic myelogenous leukaemia.

Analysis of expressed mRNAs with differential display-polymerase chain reaction (DD-PCR) is a powerful tool for the characterization of genes involved in malignant pathways and might identify markers for different phases of chronic myelogenous leukaemia (CML). We examined the presence of BCR-ABL transcripts in 25 CML patients in either the chronic phase or blast crisis. We then analysed the expression of leukocytic RNA transcripts in CML phases. DD-PCR technique was used to examine CML cases with BCR-ABL in comparison with CML cases lacking detectable BCR-ABL transcripts. Our results support the use of differential display not only for characterization of the CML differentially expressed genes but also to locate patterns that can be implemented as valuable fingerprints for each phase of CML.

Prevalence of infertility in rural areas of Kafr El-Sheikh-Egypt: a community-based study.

The current study was carried out in Desok district-Kafr El Sheikh province, to measure the prevalence of primary and secondary infertility among rural women and to study some risk factors as well. The study included 1125 married women between 15-49 years. The results of the study showed that; 7.9% reported secondary infertility 2.5% experienced primary infertility, the prevalence of primary infertility is higher among women under 30 years than older ages, and secondary infertility increases with advance in age. Both types of infertility were higher among women married under the age of 16 or above 30 years. There was an insignificant difference between fertile group and infertile groups regarding age at menarche. Irregular menses was significantly higher among infertile groups compared to fertile group. Secondary infertility group had significant higher abortion and difficult labor than fertile group. There was an insignificant difference between the study groups regarding illiteracy rate. Chronic illnesses of women as well as husbands were significantly more reported among women with secondary infertility. In conclusion, the overall prevalence of infertility is 10.4 %.

Differential display analysis of mRNAs in chronic myelogenous leukaemia

Analysis of expressed mRNAs with differential display-polymerase chain reaction [DD-PCR] is a powerful tool for the characterization of genes involved in malignant pathways and might identify markers for different phases of chronic myelogenous leukaemia [CML]. We examined the presence of BCR-ABL transcripts in 25 CML patients in either the chronic phase or blast crisis. We then analysed the expression of leukocytic RNA transcripts in CML phases. DD-PCR technique was used to examine CML cases with BCR-ABL in comparison with CML cases lacking detectable BCR-ABL transcripts. Our results support the use of differential display not only for characterization of the CML differentially expressed genes but also to locate patterns that can be implemented as valuable fingerprints for each phase of CML

DNA copy number changes in Schistosoma-associated and non-Schistosoma-associated bladder cancer.

DNA copy number changes were investigated in 69 samples of schistosoma-associated (SA) and non-schistosoma-associated (NSA) squamous cell carcinoma (SCC) and transitional cell carcinoma (TCC) of the bladder by comparative genomic hybridization (CGH). DNA copy number changes were detected in 47 tumors. SA tumors had more changes than NSA tumors (mean, 7 vs. 4), whereas the number of changes in SCC and TCC tumors was similar. SA tumors displayed more gains than losses (1.7:1), whereas NSA tumors showed an equal number of gains and losses. Changes that were observed at similar frequencies in SCC and TCC, irrespective of the schistosomal status, included gains and high-level amplifications at 1q, 8q, and 20q and losses in 9p and 13q. These changes may be involved in a common pathway for bladder tumor development and progression independent of schistosomal status or histological subtype. Losses in 3p and gains at 5p were seen only in SCC (P < 0.01) and losses in 5q were more frequent in SA-SCC than in other tumors (P < 0.05). However, changes that were more frequent in TCC than those in SCC included gains at 17q (P < 0.01) and losses in 4q (P < 0.05) and 6q (P < 0.01). Gains and high-level amplifications at 5p were seen only in SA-SCC (P < 0. 01), whereas gains and high-level amplifications with minimal common overlapping regions at 11q13 were more frequently seen both in SA-SCC and SA-TCC tumors (P < 0.01). In addition to the above mentioned alterations, several other changes were also seen at lower frequencies. The variations in the DNA copy number changes observed in TCC, SCC, SA, and NSA bladder carcinomas suggest that these tumors have different genetic pathways.

Comparison between Toxoplasma gondii DNA and specific immunoglobulins during pregnancy.

Early diagnosis of toxoplasmosis in pregnant women can be of great help in early intervention and prevention of congenital disorders that usually lead to fetal death. The purpose of the present study was to evaluate nested PCR amplification of the B1 gene of Toxoplasma gondii before and after treatment and in comparison to serological follow-up during treatment. The efficiency of treatment on the bases of PCR detection of T. gondii DNA was statistically significant, while it was insignificant when anti-toxoplasma specific IgM and IgG antibodies were used. PCR detection of T. gondii DNA when performed on whole blood is a rapid, sensitive and specific diagnostic procedure and is a valuable tool for establishing the diagnosis of T. gondii infection in women before or during pregnancy.

Screening of dystrophin gene deletions in Egyptian patients with DMD/BMD muscular dystrophies.

Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations within the dystrophin gene. Our study has identified 100 Egyptian families collected from the Human Genetics Clinic, National Research Center, Cairo. All cases were subjected to complete clinical evaluation pedigree analysis, electromyography studies, estimation of serum creatine phosphokinase enzyme (CPK) levels and DNA analysis. Multiplex PCR using 18 pairs of specific primers were used for screening of deletion mutations within the dystrophin gene. A frequency of 55% among the families. Sixty per cent of detected deletions involved multiple exons spanning the major or the minor hot spot of the dystrophin gene. The remainder 40% which mainly involved exon 45. Comparing these findings with frequencies of other countries it was found that our figures fall within the reported range of 40%- distribution of deletions in our study and other different studies was variable and specific ethnic differences do not apparently account for specific deletions. In addition this study concluded that employment of the 18 exon analysis is a cost effective and a highly accurate (97% to launch a nationwide program.