Isolation, Characterization, and Metabolic Profiling of Ceratorhiza hydrophila from the Aquatic Plant Myriophyllum spicatum

The goal of the present study was to investigate the antibacterial properties, enzyme production, and metabolic profiling of a new Ceratorhiza hydrophila strain isolated from the submerged aquatic plant Myriophyllum spicatum. Furthermore, the fungus’ morphological characterization and DNA sequencing have been described. The fungus has been identified and submitted to the GenBank as Ceratorhiza hydrophila isolate EG19 and the fungus ID is MK387081. The enzyme analyses showed its ability to produce protease and cellulase enzymes. According to the CSLI standard, the ethyl acetate extract of C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia, Micrococcus luteus, and Staphylococcus aureus. Metabolic profiling has been carried out using 700 MHz NMR spectroscopy. Based on the 1 H and 1 H- 13 C heteronuclear single quantum coherence (HSQC) NMR data and NMR databases, 23 compounds have been identified. The identified metabolites include 31% amino acids, 9% sugars, 9% amines, 4% sugar alcohols, and 4% alkaloids. This is the first report for the metabolic characterization of C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia,, which gave preliminary information about the fungus. It is expected that our findings not only will pave the way to other perspectives in enormous applications using C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia, as a new promising source of antimicrobial agents and essential metabolites, but also it will be valuable in the classification and chemotaxonomy of the species.

Molecular characteristics of methicillin-resistant Staphylococcus aureus isolates from hospital and community environments in northeastern Brazil

ABSTRACT This study characterized 30 MRSA isolates from intensive care unit (ICU) environment and equipment surfaces and healthy children. The SCCmec types I, IVa and V were detected in HA-MRSA isolates while CA-MRSA showed the SCCmec type IVa and V. Most isolates were classified as agr group II. All isolates presented the sei gene, and only HA-MRSA were positive for etb e tst genes. Three genotypes were related to Pediatric (ST5/SCCmecIV) and Berlin (ST45/SCCmecIV) clones. The present study showed molecular similarity between CA- and HA-MRSA isolates in hospital and community settings in a Brazilian region.

Estimation of the prophylactic effect of the egyptian propolis extract against aluminum silicate toxicity on some organs of albino rats growth performance and histochemical studies

Background: the liver is a critical organ because it contains most of the accumulated metals where toxic effects can expected. Also, the lung is directly affected by receiving aluminum as aluminum silicate. Exposure of aluminum leads to production of free radicals that damage living organs and tissues. Aim of the work: this study aimed to evaluate the nucleic acid content in liver, lung and cerebellum tissues intoxicated by aluminum silicate (AlS) and the possible ameliorative effect of propolis extract (PP). Material and methods: Forty male albino rats (weighting 100-120 grams) were categorized randomly into four groups, ten rats on each group (n=10). The 1st group considered as the healthy control group. The 2nd group received 200 ml PP/kg b.wt., day after day by stomach gavage. The 3rd group was injected intraperitoneally by 20 mg AlS/kg b.wt., twice weekly. The 4th group was treated with AlS in addition to PP as the same doses as in the 2nd and 3rd groups. After two months for each group. Liver, lung and cerebellum organs were harvested. Results: decreased body weight gain of rats was realized with weakly stained nucleic acids contents in liver, lung and cerebellum tissues that intoxicated by AlS. While, using the supplemented treatment (PP) at the same time with the induction of AlS compound showed an ameliorative effect on the nucleic acid contents. Conclusion: propolis has anti-oxidant by inhibiting AlS toxicity on nucleic acids in the different experimental organs of rats

Improved reverse transcription-polymerase chain reaction assay for the detection of flaviviruses with semi-nested primers for discrimination between dengue virus serotypes and Zika virus

BACKGROUND The genus Flavivirus includes a variety of medically important viruses, including dengue virus (DENV) and Zika virus (ZIKV), which are most prevalent in Brazil. Because the clinical profile of patients affected by different DENV serotypes or ZIKV may be similar, the development of new methods that facilitate a rapid and accurate diagnosis is crucial. OBJECTIVES The current study aimed to develop an improved reverse transcription-polymerase chain reaction (RT-PCR) protocol for universal detection of flaviviruses by using semi-nested primers that discriminate between DENV serotypes and ZIKV. METHODS The bioinformatics workflow adopted for primer design included: (1) alignment of 1,442 flavivirus genome sequences, (2) characterisation of 27 conserved regions, (3) generation of a primer set comprising 77 universal primers, and (4) selection of primer pairs with greatest coverage and specificity. Following primer design, the reaction was validated in vitro. The same approach was applied to the design of primers specific for DENV and ZIKV, using a species-specific sequence database. FINDINGS The new assay amplified an 800-806 nt variable region of the NS5 gene and allowed discrimination of virtually all flavivirus species using reference-sequence comparison. The 800-806 nt fragment was validated as a template for a semi-nested multiplex PCR using five additional primers for the detection of DENV and ZIKV. These primers were designed to generate amplicons of different sizes, allowing differentiation of the four serotypes of DENV, and ZIKV using agarose gel electrophoresis. MAIN CONCLUSIONS The bioinformatics pipeline allowed efficient primer design, making it possible to identify the best targets within the coding region of the NS5 protein. The multiplex system proved effective in differentiation of DENV1-4 and ZIKV on a 2% agarose gel. The possibility of discriminating DENV serotypes and ZIKV in the same reaction provided a faster result consuming less sample. In addition, this simplified approach ensured the reduction of the cost per analysis.

Community-acquired methicillin-resistant Staphylococcus aureus carrying SCCmec type IV and V isolated from healthy children attending public daycares in northeastern Brazil

Abstract Nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) have increasingly been reported in healthy communities. This study aimed to assess the rate of S. aureus in general and MRSA in particular from nasal secretion of children in daycare centers in Vitória da Conquista, Brazil. The isolates were identified based on morphology, biochemical tests and by PCR. Detection of virulence genes, biofilm production, and susceptibility test by disk diffusion agar were performed. MRSA isolates were characterized by spa, SCCmec, and multilocus sequence typing (MLST). S. aureus were recovered from 70 (47.3%) of 148 children. Among the 11 MRSA strains (15.7%), two SCCmec types (IV and V) were detected. MLST identified four STs related to three clonal complexes (CC): 5, 45, and 398. Four spa types were found circulating in this setting. Resistance of S. aureus isolates to ampicillin, erythromycin, ciprofloxacin, clindamycin, and tetracycline was 80%, 32.8%, 7.1%, 7.1% and 4.3%, respectively. One isolate presented intermediate resistance to vancomycin detected by Etest methodology. All strains were biofilm producers. The virulence genes seb, sec, spa, and pvl were detected in some isolates. This study revealed a high rate of children carrying MRSA among healthy attendees in daycare centers in Vitória da Conquista, Brazil.

efficacy of non cultured melanocytes and keratinocytes transplantation in treatment of generalized vitiligo

Objective: determine the efficacy and safety of the non-cultured melanocyte-keratinocyte transplantation procedure in the treatment of stable resistant generalized vitiligo. Design: A simpler and modified method based on that of Olsson and Juhlin [1998] was performed this method uses a shaved biopsy skin sample one tenth the size of the recipient area. The skin sample is incubated, and the cells are mechanically separated using trypsin-EDTA solution and then centrifuged to prepare a suspension. Cell suspension is then applied to the derma braded depigmented skin area, and a collagen dressing is applied to keep it in place

Patients: twenty one patients with stable generalized vitiligo of different sites and sizes were treated with NMKT and followed up for 6 months

Intervention: Autologous, non-cultured melanocyte keratinocyte cell transplantation

Main Outcome Measure: initial pigmentation was observed. Regimentation was graded as excellent with 75 to 100% pigmentation, good with 50% to 74% fair with 25 to 49%, and poor with <25% pigmentation, assessment by modified vitiligo area scoring index, color match ,patient satisfaction and adverse events were assessed

Results: 9.66% showed excellent response, 11.50 % showed good response, 40.38% showed fair response and 38.46 % with poor response. Average percent change in VASI was 24.56% +-33.71. The color matching was excellent to good in 65.4% and poor in 34.6% of lesions, 3.8% of patients only were very satisfied and complications were minimal. Limitations: Limitations include small sample size, lack of control group and short follow-up period

Conclusions: NMKT is an easy economic technique, which may be used in resistant areas of stable vitiligo. The smaller the size of the lesion and the longer the stability duration the higher the percentage of regimentation response obtained. Results tend to be better over the trunk and proximal limbs than elbows, knees and distal extremities. Complications are minimal the most common is post inflammatory hyperpigmentation of the donor area

Cardiac involvement in Sudanese children with connective tissue disease

Connective tissue diseases [CTD] are associated with a variable range of cardiac abnormalities. We identified all patients with CTD and cardiac involvement seen at Jafar Ibn Ouf Children's Hospital and the Sudan Heart Centre by one cardiologist between Jan 2005 and January 2010. Clinical and echocardiographic evaluations were done and arranged consultation with rheumatologist. Nine patients were identified, three males and six females. Five patients [62%] were initially seen by the cardiologist and CTD was suspected by clinical examination in four patients and in one patient diagnosed by routine screening during workup for pulmonary hypertension. Symptoms were present for one month to 3 years before presentation to the cardiologist. In three patients [30%] the cardiac symptoms preceded the rheumatological symptoms. In one patient, aortic valve replacement for presumed rheumatic aortic regurgitation preceded arthritis by 12 months. Patients with CTD may present first to the cardiologist with findings that mimic common cardiac lesions. All patients with CTD should have routine evaluation by the cardiologist so as to implement optimal early interventions

Fatal abdominal sarcomatosis secondary to gastrointestinal stromal tumor with bland histology

Fatal abdominal sarcomatosis is a rare complication secondary to gastrointestinal stromal tumor [GIST] arising from the small intestine. Here, we describe a 49-year-old man who presented with massive ascites and multiple mesenteric solid masses. Autopsy showed large necrotic mass [19x14x8.0 cm] surrounding the terminal ileum with multiple smaller nodules on the omentum and mesentery. Histological examination revealed highly cellular, predominantly bland, spindle cells with low mitotic activity. Gastrointestinal stromal tumor was confirmed by strong positive staining for c-KIT [CD 117]. Our case is an example of fatal intra- abdominal sarcomatosis of GIST where the histological features were essentially bland

Fetal macrosomia greater than or equal to 4000 grains. Comparing maternal and neonatal outcomes in diabetic and nondiabetic women

To compare maternal and neonatal outcomes of fetal macrosomia in diabetic and nondiabetic women. A retrospective case-control study was conducted at Riyadh Military Hospital, Riyadh, Kingdom of Saudi Arabia from January 2004 to December 2006. A total of 766 macrosomic newborns met the inclusion criteria. We compared maternal and neonatal characteristics and outcomes between diabetic [group 1, n=207] and nondiabetic [group 2, n=559] women. There were significantly more macrosomic newborns in nondiabetic women; 73% versus 27% in diabetic women, p=0.0001. Cesarean deliveries were significantly higher in diabetic women compared to nondiabetic women [30.4% versus 19.5%, p=0.002]. There were no significant differences between either group in total maternal morbidity [12.6% versus 14.1%, p=0.7]. There were significantly more severe cases of shoulder dystocia occurring in newborns of diabetic women compared to nondiabetic women [1.9% versus 0.2%, p=0.03]. Elective cesarean delivery for estimated fetal weight >/= 4500g for nondiabetic women and >/= 4250 g for diabetic women may avoid severe shoulder dystocia without increasing maternal morbidity rates

Rapid detection of Haemophilus influenzae type b in Bangladeshi children with pneumonia and meningitis by PCR and analysis of antimicrobial resistance.

A polymerase chain reaction (PCR) assay with primers from 'bexA' gene was compared with culture for the detection of Haemophilus influenzae type b (Hib) in clinical samples from children with pneumonia and meningitis. Of 200 sera (180 from pneumonia, 20 from non-pneumonia patients) tested by PCR (serum-PCR), Hib was detected in 15 of 16 blood culture-positive and in 6 blood culture-negative pneumonia cases. When compared with the results of blood culture, serum-PCR had sensitivity, specificity, and accuracy index of 93.7%, 96.7%, and 96.5% respectively. Of 120 cerebrospinal fluid (CSF) samples from meningitis patients tested by culture and PCR (CSF-PCR), the latter method could detect Hib in all 15 culture-positive and in 8 of 105 culture-negative cases, showing sensitivity, specificity, and accuracy index of 100%, 92.4%, and 94.4% respectively. The PCR result was available within a day. Antimicrobial susceptibility of Hib was determined by the disc-diffusion method. High rate of resistance to ampicillin (54.8%), chloramphenicol (48.4%), and co-trimoxazole (80.6%) was observed among 31 invasive Hib isolates with resistance to all 3 drugs (multiresistance) in 48.4% of the isolates. All the Hib isolates were susceptible to ceftriaxone. The study has shown that PCR is a rapid, sensitive and specific diagnostic test for Hib from clinical samples, and a combination of culture and PCR is necessary for the detection of Hib infections to the maximum extent for case management to reduce morbidity, mortality, and complications of the invasive Hib infections. A high prevalence of multiresistant Hib strains is a matter of concern.

Seroprevalence of hepatitis B, hepatitis C, CMV and HIV in multiply transfused thalassemia patients: results from a thalassemia day care center in Malaysia.

Regular blood transfusions for patients with thalassemia have improved their overall survival although these transfusions carry a definite risk of the transmission of certain viruses. Infection with hepatitis B virus (HBV), hepatitis C virus (HCV), cytomegalovirus (CMV) and human immunodeficiency virus (HIV) leads to complications which contribute to the morbidity and mortality of patients with thalassemia. We analyzed the blood samples taken from 85 transfusion dependent thalassemics receiving treatment at the day care center in Hospital Universiti Kebangsaan Malaysia and found that the seroprevalence rates for HBV, HCV and CMV were 2.4%, 22.4% and 91.8% respectively. None of the patients tested positive for HIV. Those positive for HBV and HCV will require further tests and treatment if chronic hepatitis is confirmed.

Development and evaluation of co-agglutination test to detect rotavirus antigens in stools of patients with diarrhoea.

Rotavirus is the most common cause of acute diarrhoea in infants and children in both the developed and developing countries including Bangladesh. Information about rotavirus diarrhoea in Bangladesh is insufficient primarily due to the lack of diagnostic facilities due to the high cost of reagents and equipment and lack of skilled personnel. A simple, suitable and less costly technique of co-agglutination test using protein-A secreting staphylococci was developed and evaluated against a commercially available ELISA kit to detect rotavirus antigen in stools of patients with diarrhoea. Staphylococcus cowan strain 1 was grown and coated with rabbit antisera raised against RV5 and SA11 rotavirus strains. The antibody-coated staphylococci were agglutinated specifically by rotavirus present in faecal samples within one or two minutes. A total of 1332 stool specimens were tested by co-agglutination and ELISA, of which 210 (15.77%) were positive by ELISA and 276 (20.72%) by co-agglutination test. Compared to ELISA, sensitivity of co-agglutination test was 76.19%, specificity 89.66% and predictive values of a positive and a negative test were 57.97% and 95.26% respectively. The results indicate that the co-agglutination test is a simple and suitable technique for rapid screening of rotavirus infection which could be adopted in clinical practice.

Epidemiological study and comparison of pyrantel and levamisole in the treatment of roundworm and hookworm infestations.

Stool examined for presence of nematodes from 467 school students in Dacca city showed 48% ova positive cases with 22% among them cases of mixed nematode infection. Highest prevalence of 58% was observed with roundworm and 5% was hookworm infestations. Among hookworm cases, Necator americana (68%) appears to be the predominant species. Two anthelmintics-pyrantel and levamisole achieved similar high cure rate (around 97%) and Egg reduction rate of 99% in roundworm infection. However, pyrantel was observed to be more effective in hookworm infestation (87% against 60%) particularly in Necator americana cases.