Detection of Antibiotic Resistance Genes (CTX-M, Van A and Van B) of Enterococcus faecalis Isolated from Children with Bacteremia by RT-PCR.

Fever is one of the most common diseases affecting humans, as it results from any disease or development and worsening of the disease for most people with widespread infections in the body. Therefore, this study aimed to evaluate antibiotic resistance genes (CTX-M, Van A and Van B) of Enterococcus faecalis isolated from children with bacteremia by RT-PCR. A total of 200 children was enrolled in the study, 100 children with fever and 100 healthy children (not suffering from any problem); that is, they are a control group for the detection of antibiotic resistance genes (CTX-M, Van A and Van B) of Enterococcus faecalis by RT-PCR. The age of the two groups ranged from one to five years. Four ml of venous blood sample was collected from each child; the venipuncture area was sterilized first with alcohol at a rate of 70%, followed by medical iodine and then sterilized with alcohol again to avoid contamination with skin flora. The blood samples were cultured on media for isolating bacteria. Then, the resistant isolates of E. faecalis to Vancomycin and cefotaxime antibiotics were taken and kept in special nutrient agar media where the DNA of the bacteria was extracted using (Zymogene Extraction kit, Japan). The detection of the exact genes (CTX-M, Van A and Van B) was done using Real-Time PCR technology according to the protocol mentioned by the company (Sacace biotechnology, Italy). The study presented that 40% of children with fever have positive blood cultures compared with 5% in the control group, with a significant difference between the two groups (P<0.001). The study found that 32.5% of bacteremic children were due to S. aureus, 30%, 5%, and 4% were due to E. faecalis, E. coli, P. aeruginosa and Klebsiella spp, respectively, with significant difference (P<0.01). The study showed that 91.67% of E. faecalis isolates were sensitive to Levofloxacin, 83.33% to Amoxiclav, 66.67% to Erythromycin, 58.33% to Amikacin, 50% to Ampicillin, 33.33% to cefotaxime and Ceftriaxone and 25% toward Vancomycin. From 9 isolates resistant to Vancomycin, the study presented that 88.89% of them were observed with Van A gene production as detected by real-time PCR (P<0.001). The study also showed that 77.78% were observed with Van B gene production as detected by real-time PCR (P<0.001). The study revealed that all E. faecalis isolates resistant to cefotaxime and Ceftriaxone were characterized by CTX gene production as detected by real-time PCR (P<0.001).

Phenolic profiling of an extract from Eugenia jambos L. (Alston)--the structure of three flavonoid glycosides--antioxidant and cytotoxic activities.

Phenolic metabolite profiling and identification using high performance liquid chromatography (HPLC) coupled to high resolution accurate mass spectrometry (HR-ESI-MS) with detection of negative ions was used for assaying the complex mixture of phenolics of an aqueous ethanol leaf extract of Eugeniajambos L. (Myrtaceae). Eight known polyphenolics were tentatively identified, and, in addition, three hitherto unknown flavonol-O-glycosides were detected in the extract. These unknowns were taken as the targets and isolated by means of consecutive polyamide S6, MCI gel and repeated Sephadex LH-20 column fractionation. The isolation and purification were monitored by HPLC/ESI-MS. The isolates were subsequently identified as quercetin 3-O-xylosyl-(1"' --> 2")-O-xyloside, myricetin 7-methylether 3-O-xylosyl-(1"' --> 2")-rhamnoside and myricetin 3',5'-dimethyl ether 3-O-xylosyl-(1"'-->* 2")-O-rhamnoside. All known metabolites were also separated by applying the same chromatographic techniques. ESI-MS, ¹H and ¹³C NMR spectra were then recorded, completely interpreted and confirmed by HR-ESI-MS and 2D NMR spectroscopy. In order to get information about biological activities of E. jambos the extract was tested for radical scavenging activity by DPPH and ORAC assay. In addition, its cytotoxicity was assessed by the neutral red assay against non-tumorigenic HaCaT keratinocytes and the human bladder carcinoma cell line 5637.

Phenolics from Caesalpinia ferrea Mart.: antioxidant, cytotoxic and hypolipidemic activity.

Nine phenolics were isolated from the aqueous ethanol extract of the leaves of Caesalpinia ferrea. The isolates were characterized for the first time from that plant. The structures of all isolates (1-9) were elucidated by conventional methods, spectroscopic analysis, including 1 D and 2D NMR, and by HR-ESIMS as well. The antioxidant capacities using the ORAC method and the cytotoxic activity using the neutral red assay (NRU) for that extract and three major isolates have been evaluated. In addition, the hypolipidemic activity (in vitro and in vivo) of the extract has been assessed.

Identification of Two Homozygous Sequence Variants in the COL7A1 Gene Underlying Dystrophic Epidermolysis Bullosa by Whole-Exome Analysis in a Consanguineous Family.

Dystrophic epidermolysis bullosa (DEB) is an inherited skin disorder with variable severity and heterogeneous genetic involvement. Diagnostic approaches for this condition include clinical evaluations and electron microscopy of patients' skin biopsies, followed by Sanger sequencing (SS) of a large gene (118 exons) that encodes the alpha chain of type VII collagen (COL7A1) located on Chromosome 3p21.1. However, the use of SS may hinder diagnostic efficiency and lead to delays because it is costly and time-consuming. We evaluated a 5-generation consanguineous family with 3 affected individuals presenting the severe generalised DEB phenotype. Human whole-exome sequencing (WES) revealed 2 homozygous sequence variants: the previously reported variant p.Arg578* in exon 13 and a novel variant p.Arg2063Gln in exon 74 of the COL7A1 gene. Validation by SS, performed on all family members, confirmed the cosegregation of the 2 variants with the disease phenotype. To the best of our knowledge, 2 homozygous COL7A1 variants have never been simultaneously reported in DEB patients; however, the upstream protein truncation variant is more likely to be disease-causing than the novel missense variant. WES can be used as an efficient molecular diagnostic tool for evaluating autosomal recessive forms of DEB.

Case of Sjögren-Larsson syndrome with a large deletion in the ALDH3A2 gene confirmed by single nucleotide polymorphism array analysis.

Sjögren-Larsson syndrome (SLS) is a neurocutaneous disorder inherited in an autosomal recessive fashion. SLS patients are characterized by lipid metabolism error, primarily leading to cardinal signs of ichthyosis, spasticity and mental retardation. Additional signs include short stature, epilepsy, retinal abnormalities and photophobia. More than 90 mutations of the ALDH3A2 gene have been reported for SLS, and such variants can be successfully detected at a rate of 94% by direct DNA sequencing. We performed direct sequencing of ALDH3A2 gene from the index patient, however, no mutation could be detected. HumanCytoSNPs12 array analysis and subsequent targeted single nucleotide polymorphism analysis revealed a novel deletion mutation at chromosome 17p11.2. This 67-Kb region includes the first five coding exons of ALDH3A2, and is flanked by rs2245639 and rs962801. To the best of our knowledge, this mutation is novel and our findings broaden the mutation spectrum of ALDH3A2 causing SLS phenotype.

Soft tissue high grade myoepithelial carcinoma with round cell morphology: report of a newly described entity with EWSR1 gene rearrangement.

The case of soft tissue malignant myoepithelioma is presented including clinicopathological, immunohistochemical and cytogenetic findings. A 36-year-old Saudi male patient suffered from large mass involving right scapula and right shoulder joint measuring 14x13x11 mm. Core biopsy revealed sheets and lobules of poorly differentiated small malignant cells with marked atypia and frequent mitosis. Initially, immunohistochemistry was reactive for vimentin, pan-cytokeratin, EMA and CD99. The case was negative for desmin, SMA, CD34, S-100 protein and GFAP. FISH analysis exhibited negativity for SS18 (18q11.2) gene rearrangement and positivity for EWSR1 (22q12) gene rearrangement and a diagnosis of Ewing/PNET was considered. Clinical behavior and therapeutic response did not match the diagnosis with re-evaluation. Wedge biopsy demonstrated aggregates of epithelioid cells besides calponin and P63 positivity. Final diagnosis of malignant myoepithelioma with EWSR1 gene rearrangement was issued; a new entity with aggressive course. Myoepithelial carcinoma of soft tissue exhibits a wide spectrum of cytomorphology with overlapping phenotype similar to other soft tissue sarcoma like synovial sarcoma, mesenchymal chondrosarcoma, epithelioid sarcoma as well as Ewing/PNET. Moreover, a new finding of EWSR1 gene rearrangement is recognized in malignant myoepithelioma with different fusion partners. Hence, myoepithelial carcinoma should be kept in mind in diagnosis of soft tissue tumors even with unusual phenotype and gene rearrangement.

Aphyllin, the first isoferulic acid glycoside and other phenolics from Tamarix aphylla flowers.

The first glycosylated isoferulic acid, isoferulic acid 3-O-beta-glucopyranoside, together with the new phenolics, tamarixetin 3,3'-di-sodium sulphate and dehydrodigallic acid dimetyl ester have been characterized from a flower extract of Tamarix aphylla. The structures were established on the basis of spectral data. The extract exhibited a distinct radical scavenging effect and to improve the viability of human keratinocytes (HaCaT cells). Also, the known isoferulic acid and ferulic acid which have been determined to be the major components of the investigated extract by HPLC/ESI mass spectrometric screening have been separated, characterized and evaluated as active antioxidants and as cell activity stimulating agents as well.

Bone mineralization enhancing activity of a methoxyellagic acid glucoside from a Feijoa sellowiana leaf extract.

The capability of an aqueous methanol extract obtained from the leaves of Feijoa sellowiana Berg. on possible prevention and treatment of osteoporosis has been examined by evaluating its stimulating effect on the two human osteoblastic cell lines HOS58 and SaOS-2. The extract was found to increase significantly the mineralization of cultivated human bone cell, whereby a clear increment (15.3 +/- 2.7%) in von Kossa positive area was determined when administering 25 microg/ml leaf extract. A phytochemical investigation of the extract has demonstrated the high phenolic content and led to the isolation and identification of twenty three of them, among which the new 3-methoxyellagic acid 4-O-beta-glucopyranoside was fully identified. All structures were elucidated on the basis of conventional analytical methods and confirmed by FTMS, 1D- and 2D-NMR data. The new compound was found to cause a significant increase of mineralized area at 20 microg/mL, while at lower concentrations the effect was not significant. However, an increase of the number of mineralized spots (nodules) at all tested concentrations of the compound was observed.

Ericifolin: an eugenol 5-O-galloylglucoside and other phenolics from Melaleuca ericifolia.

Ericifolin, an eugenol 5-O-beta-(6'-O-galloylglucopyranoside) possessing the naturally unknown phenolic moiety, 5-hydroxyeugenol, together with the two new phenolics, 2-O-p-hydroxybenzoyl-6-O-galloyl-(alpha/beta)-4C1-glucopyranose and 3-methoxyellagic acid 4-O-rhamnopyranoside have been isolated from the antibacterial leaves extract of Melaleuca ericifolia. In addition, 19 known phenolics were also separated and characterized. All structures were elucidated on the basis of analysis of 1H, 13C NMR, HMQC, HMBC and FTMS spectral data.

BDN programmes and the effect of medical students' interventions to promote child health in Sudan.

Basic development needs (BDN) is an approach to community development that aims at improving quality of life through the full involvement and self-management of communities, supported by intersectoral collaboration. The objective of this study was to assess the effects of BDN implementation in an area of Sudan on aspects of a programme implemented by students from the Faculty of Medicine, University of Gezira. The study revealed that the joint implementation of BDN programmes and students' interventions brought about marked improvement of family and community practices as well as greater reduction in the incidence of all assessed major childhood illnesses (diarrhoea, cough and fever) than when the students' programme was implemented alone. BDN implementation also facilitated the retrieval of good quality community-based data.

BDN programmes and the effect of medical students' interventions to promote child health in Sudan

Basic development needs [BDN] is an approach to community development that aims at improving quality of life through the full involvement and self-management of communities, supported by intersectoral collaboration. The objective of this study was to assess the effects of BDN implementation in an area of Sudan on aspects of a programme implemented by students from the Faculty of Medicine, University of Gezira. The study revealed that the joint implementation of BDN programmes and students' interventions brought about marked improvement of family and community practices as well as greater reduction in the incidence of all assessed major childhood illnesses [diarrhoea, cough and fever] than when the students' programme was implemented alone. BDN implementation also facilitated the retrieval of good quality community-based data

Phenolics from extracts of Brahea armata with inhibitory effect against 5alpha-reductase type-II.

Fractions of the aqueous alcohol extracts of the rind and kernel of Brahea aramata fruits have been investigated for their activity against 5alpha-reductase type II, which is expressed predominantly in the prostate. This isozyme represents a major target for drugs against benign prostate hyperplasia (BPH) and prostate cancer. Also, a structural analysis of the phytophenolics, present in both aqueous alcohol extracts as the major constituents, has led to the isolation of five phenolics, including the new natural product, 4',6'-dimethoxy beta,4,2'-trihydroxy chalcone from the rind extract and three phenolics, including the new natural product, 1-p-hydroxybenzoyl glycerol from the kernel extract. All structures were confirmed by ESI-MS and NMR analysis.

The economic impact of incorporating flexible endoscopy into a community general surgery practice.

BACKGROUND: Flexible endoscopy is a vital component of gastrointestinal surgery. It has and will replace many of the surgical procedures now commonly performed. Flexible endoscopy, unfortunately, is not an integral part of surgical residency training based on resident operative experience as reported by the Residency Review Committee. Moreover, general surgeons have deferred the practice of flexible endoscopy to the gastroenterologists because of concerns over turf battles and referral patterns. The purpose of this study was to assess the overall case load and the economic impact of flexible endoscopy on the practice of general surgery in a community hospital setting. METHODS: This retrospective review was performed over a 6-month period. The total cases and the total billings of inpatient and outpatient procedures for a group practice of five general surgeons in a community hospital were evaluated. The billings were the actual charges based on current procedural terminology (CPT) codes for these procedures using the Medicare fee schedule. RESULTS: Of the 2,159 procedures performed, 1,154 involved flexible endoscopy cases accounting for 54% of all cases (1,154 of 2,159) performed from February 1, 2003 to July 31, 2003. Flexible endocopy accounted for 43% of the total charges. A. total of 46 surgical procedures and 216 future endoscopies were generated from the flexible endoscopic procedures. Future endoscopic cases were for surveillance of colonic neoplasia and Barrett's esophagus. CONCLUSIONS: Flexible endoscopy contributed to a major portion of the caseload and revenue generated by the general surgery group studied. The overall impact of flexible endoscopy is even greater than reported because of the future endoscopic surveillance cases or surgical interventions generated on the basis of endoscopic findings.

Flavonoid and methoxyellagic acid sodium sulphates from Frankenia laevis L.

Two new flavonol di-sodium sulphates and an ellagic acid methyl ether mono-sodium sulphate have been found in the aqueous alcohol whole plant extract of Frankenia laevis L. They have been identified as the 3,7-di-sodium sulphate of kaempferol, the 3,7-di-sodium sulphates of quercetin and the 4'-mono-sodium sulphate of ellagic acid-3-methyl ether. Also, five known compounds have been isolated and characterized from the same extract. Establishment of all structures has been achieved mainly, by ESI-MS and NMR.

Phenolic sodium sulphates of Frankenia laevis L.

Four new phenolic anionic conjugates have been isolated from the whole plant aqueous alcohol extract of Frankenia laevis L. Their structures were established, mainly on the basis of ESI-MS, 1D and 2D NMR spectroscopic evidence, as gallic acid-3-methyl ether-5-sodium sulphate, acetophenone-4-methyl ether-2-sodium sulphate, ellagic acid-3,3'-dimethyl ether-4,4'-di-sodium sulphate and ellagic acid-3-methyl ether-4-sodium sulphate.

Unusual phenolic glycosides from Cotoneaster orbicularis.

The whole plant of Cotoneaster orbicularis contains the novel di-C-glycosylflavone, 4",4"'-di-O-beta-glucopyranosyl-vicenin II, or 6,8-di-C-beta-Cellobiosylapigenin, as well as the hitherto unknown natural phenolic glucoside, gentisic acid 2-O-beta-glucopyranoside, or orbicularin. Further phenolics are protocatechuic, anisic, caffeic, p-coumaric acids, catechin, epicatechin, 2"-O-alpha-rhamnopyranosylvitexin, vitexin, rutin, isoquercetrin, hyperin and naringenin. All structures were determined by routine methods of analysis and confirmed mostly by 1H- and 13C-NMR.

Teucrol, a decarboxyrosmarinic acid and its 4'-O-triglycoside, teucroside from Teucrium pilosum.

Teucrol, a decarboxy rosmarinic acid and its triglycoside, teucroside, 9'-decarboxyrosmarinic acid-4'-O-alpha-rhamnosyl-(1'''-->6''')-O-beta-galactosyl-(1'''-->4'')-O-alpha-rhamnoside, are two new natural phenolics which have been isolated and identified from the aqueous alcohol extract of the whole plant of Teucrium pilosum (Decne) Asch. & Schweinf. (Lamiaceae). Structures were determined by conventional methods of analysis and confirmed by MS and NMR spectral analysis, including 2D-homonuclear COSY experiment.

Metabolic changes concerning the effect of castration on some blood constituents in male rabbits.

This experiment was conducted to determine the effects of castration on some metabolic alterations concerning proteins, lipids and minerals metabolism in both immature and mature rabbits. Twenty male New Zealand White rabbits (10 immature and 10 mature) were used. Blood samples were collected from the immature and mature intact animals (control non castrated). Moreover, the castrated animal groups (immature and mature) were sampled at 2, 4, 6 and 8 weeks after castration. The obtained results revealed that, castration caused a significant decrease in serum total protein, albumin, alpha 1-globulins and alpha 2-globulins concentrations. Meanwhile, gamma-globulins level was decreased temporarily at 2 weeks after castration in mature rabbits. beta-globulins level revealed non significant decrease at 2 weeks after castration in immature rabbits followed by a non significant increase. In mature rabbits beta-globulins level non significantly increased. A significant decrease in serum total nucleic acid concentrations was observed after castration throughout the experimental periods, whereas the value of serum uric acid level markedly increased after castration. Serum lipids concentrations (total lipids, total cholesterol, phospholipids and non esterified fatty acids) were increased after castration. The value of serum trace elements including copper, iron, zinc and manganese concentrations showed a significant decrease after castration. The decrease in serum copper and zinc levels were markedly observed in mature castrated rabbits. Also, serum sodium and potassium concentrations were decreased after castration. Testosterone propionate administration in mature castrated rabbits restored most of the investigated serum blood parameters to the levels of intact non castrated mature rabbit. It could be concluded that castration (deficiency of testosterone) induced marked decrease in protein synthesis and increased protein degradation, but serum lipids markedly elevated after castration in both immature and mature animals. Also, castration caused a marked decrease in some serum minerals and trace elements.

Plasma concentrations of lipids and lipoproteins in newborn kids and female Baladi goats during late pregnancy and onset of lactation.

Concentrations of blood lipids and some lipoproteins were investigated in normal female Baladi goats during late pregnancy, parturition and onset of lactation as well as in their newborn kids during the first two weeks of life. A total number of 60 herparinized blood samples was collected from does at 4, 3, 2 and 1 weeks pre-partum, day of parturition and at 1, 2, 3 and 4 weeks postpartum. In addition, blood samples were also collected from their newborn kids during the first two weeks of life (day of birth, 1 and 2 weeks of age). Plasma was separated and analyzed for concentration of total lipid, total cholesterol, triacylglycerols, phospholipids, non esterified fatty acids (NEFA) and some lipoproteins as high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C). The obtained results revealed that there was a significant decrease in plasma level of total lipids at one week after parturition. Plasma level of triaclyglycerols was significantly higher at 4, 3 and 2 weeks before parturition. This increase became very highly significant at one week before parturition. Meanwhile, plasma phospholipid concentrations showed a significant decrease at 3 weeks before parturition, followed by an significant increase at 2 and 3 weeks after parturition and highly significant increase at 4 weeks after parturition. The concentration of plasma NEFA showed a significant increase at 4 weeks before parturition followed by a very highly significant increase at 2 and 1 week before parturition. On the other hand plasma NEFA was non detected at 2, 3 and 4 weeks post-partum when compared with the value reported at day of parturition. Regarding plasma lipoprotein concentrations the obtained results showed that there was a significant increase in plasma HDL-C level at 2 and 3 weeks after parturition, followed by a very highly significant decrease at the fourth week post-partum. However, plasma LDL-C level showed a significant decrease at 3, 2 and 1 weeks before parturition, followed by a further highly significant decrease at 1 and 2 weeks post-partum. In newborn kids plasma concentrations of total lipids, total cholesterol, phospholipids, HDL-C and LDL-C were very markedly increased at 1 and 2 weeks of age. However, plasma triacylglycerol concentrations showed a highly significant decrease at 1 and 2 weeks of age. The concentration of plasma NEFA showed a very highly significant decrease at 2 weeks of age, whereas, at one week of age plasma NEFA were not detected in comparison with first day of life.