Differential expression of CYP2j2 gene and protein in Camelus dromedarius.

CYP2J2 is a member of the cytochrome P450 superfamily. It had been described in different mammalian species; however, no studies have described this gene in Camelus dromedarius. CYP2J2 is an epoxygenase enzyme which oxidizes various fatty acids, mainly arachidonic acid, via NADPH-dependent epoxidation to generate epoxyeicosatrienoic acids (EETs). It is a multi-functional enzyme that plays crucial roles in inflammation, cancer, drug metabolism, and embryo development. It controls the water re-absorption in the kidney and maintains the blood pressure and glucose homeostasis. This study is considered the first report investigating the differential expression profiles of the CYP2J2 mRNA and protein in the liver, heart, and kidney of Camelus dromedarius. A total of 30 samples were used to determine the expression of both CYP2J2 mRNA and protein using qRT-PCR and western blotting methods, respectively. The mRNA level of CYP2J2 was significantly elevated in the liver compared to that in the heart and kidney. The tissue distribution of the CYP2J2 protein was coherent to its transcript level in the kidney, but not in the liver and heart samples. The difference between the CYP2J2 mRNA and protein distributions in the three studied organs may be attributed to the mechanism by which the CYP2J2 might be involved in the adaptability of the camel to the arid environment.

Epidemiology and public health significance of Cryptosporidium isolated from cattle, buffaloes, and humans in Egypt.

The epidemiology and public health significance of Cryptosporidium species and genotypes were investigated in Beni-Suef Governorate, Egypt. A total of 610 animal fecal samples (480 from cattle and 130 from buffaloes) beside 290 stool samples from humans were collected in the period between January and December 2014. Based on the microscopic examination, the overall estimated prevalence of Cryptosporidium spp. in cattle, buffaloes, and humans was 10.2, 12.3, and 19 %, respectively. The highest detection rates were in calves less than 2 months of age (17.1 %) and diarrheic animals (13.0 %). Likewise in humans, the highest prevalence of Cryptosporidium was in infants (31.3 %) and diarrheic individuals (21.1 %). The gender distribution in humans denoted that Cryptosporidium was reported more frequently in males (21.7 %) than females (14.5 %). Based on the molecular characterization of Cryptosporidium, Cryptosporidium oocyst wall protein (COWP) and gp60 genes were successfully amplified in 36 out of 50 samples subjected to genotyping. Restriction fragment length polymorphism (RFLP) analysis of the COWP fragments revealed that Cryptosporidium parvum was the only species detected in cattle (12 isolates) and buffaloes (4 isolates), while in humans, the detected species were Cryptosporidium hominis (15 isolates) and C. parvum (5 isolates). Sequence analysis of the gp60 gene identified the subtype IIdA20G1 within C. parvum isolated from both animals and humans. The common occurrence of zoonotic subtypes of C. parvum in cattle and buffaloes highlights the potential role of these animals as significant reservoirs of infection to humans. Also, the presence of C. hominis and C. parvum in humans indicates that both anthroponotic and zoonotic pathways are expected.

MH17: the Malaysian experience.

A disaster is a natural or man-made (or technological) hazard resulting in an event of substantial extent causing significant physical damage or destruction, loss of life, or drastic change to the environment. It is a phenomenon that can cause damage to life and property and destroy the economic, social and cultural life of the people; and overwhelms the capacity of the community to cope with the event. The recent tragic aviation accidents in 2014 involving Malaysia Airlines flights MH370 and MH17 shocked the world in an unprecedented manner. This paper focuses on the Malaysian experience in the MH17 mission in Ukraine as well as the first ever international Disaster Victim Identification (DVI) operation for the Malaysian DVI team. The DVI operations in Hilversum, the Netherlands were well described in stages. The Netherlands' Landelijk Team Forensische Opsporing as the lead DVI team in Hilversum operated systematically, ensuring the success of the whole mission. This paper discusses the lessons learned by the Malaysian team on proper DVI structure, inter- and intra-agency cooperation, facilities planning and set up, logistics and health and safety aspects, as well as effective communication and collaboration with other international delegates. Several issues and challenges faced by the Malaysian team were also documented. In addition, the authors shared views, opinions and recommendations for a more comprehensive DVI operation in the future.

Significant impact of IL-6 -174G/C but inverse relation with -634 C/G polymorphism in patients with non-small cell lung cancer in Kashmiri population.

To study the possible role of proinflammatory interleukin 6 -174 G>C (rs 1800795) and -634 C>G (rs 1800796) polymorphism in the pathogenesis of non-small cell lung cancer (NSCLC). A total of 190 NSCLC patients and 200 healthy controls were evaluated for polymorphic analysis of -174 G/C and -634 C/G by PCR-RFLP followed by DNA sequencing. A significant association was observed in the genotypic and allelic distribution of IL-6 -174 G/C in the NSCLC group as compared to control group [OR = 2.7 (1.77-4.11), p < 0.0001]. Smokers with the -174C allele were found to be significantly associated with NSCLC (p = 0.01), while 634C/G SNP showed an inverse relation [OR-0.4, p < 0.0001]. The present investigation revealed a significant association of the IL6 -174 G/C gene promoter polymorphism with NSCLC, and thus, the IL-6 -174G/C genotype can be considered as one of the biological markers in the etiology of NSCLC.

c-Met is a prognostic marker and potential therapeutic target in clear cell renal cell carcinoma.

BACKGROUND: Activation of the c-Met pathway occurs in a range of malignancies, including papillary renal cell carcinoma (RCC). Its activity in clear cell RCC is less clear. We investigated c-Met expression and inhibition in a large cohort of RCC tumors and cell lines. METHODS: c-Met protein expression was determined by automated quantitative analysis (AQUA) on a tissue microarray (TMA) constructed from 330 RCC tumors paired with adjacent normal renal tissue. c-Met expression and selective inhibition with SU11274 and ARQ 197 were studied in clear cell RCC cell lines. RESULTS: Higher c-Met expression was detected in all RCC subtypes than in the adjacent normal renal tissue (P < 0.0001). Expression was highest in papillary and sarcomatoid subtypes, and high-grade and stage tumors. Higher c-Met expression correlated with worse disease-specific survival [risk ratio = 1.36; 95% confidence interval (CI) 1.08-1.74; P = 0.0091] and was an independent predictor of survival, maintained in clear cell subset analyses. c-Met protein was activated in all cell lines, and proliferation (and colony formation) was blocked by SU11274 and ARQ 197. CONCLUSIONS: c-Met is associated with poor pathologic features and prognosis in RCC. c-Met inhibition demonstrates in vitro activity against clear cell RCC. Further study of ARQ 197 with appropriate biomarker studies in RCC is warranted.

Characterization and risk factors of vancomycin-resistant Enterococci (VRE) among animal-affiliated workers in Malaysia.

AIMS: This study determined the risk factors and characteristics of vancomycin-resistant Enterococci (VRE) among individuals working with animals in Malaysia. METHODS AND RESULTS: Targeted cross-sectional studies accompanied with laboratory analysis for the identification and characterization of resistance and virulence genes and with genotype of VRE were performed. VRE were detected in 9·4% (95% CI: 6·46-13·12) of the sampled populations. Enterococcus faecium, Enterococcus faecalis and Enterococcus gallinarum were isolated, and vanA was detected in 70% of the isolates. Enterococcus faecalis with vanB was obtained from one foreign poultry worker. At least one virulence gene was detected in >50% of Ent. faecium and Ent. faecalis isolates. The esp and gelE genes were common among Ent. faecium (58·3%) and Ent. faecalis (78%), respectively. The VRE species showed diverse RAPD profiles with some clustering of strains based on the individual's background. However, the risk factors found to be significantly associated with the prevalence of VRE were age (OR: 5·39, 95% CI: 1·98-14·61) and previous hospitalization (OR: 4·06, 95% CI: 1·33-12·35). CONCLUSION: VRE species isolated from individuals in this study have high level of vancomycin resistance, were genetically diverse and possessed the virulence traits. Age of individuals and history of hospitalization rather than occupational background determined VRE colonization. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides comprehensive findings on the epidemiological and molecular features of VRE among healthy individuals working with animals.

Significant Psychosocial Influence in Frail People Living with HIV Independent of Frailty Instrument Used.

BACKGROUND: Antiretroviral therapy (ART) usage among people living with HIV (PLWH) has led to significant mortality declines and increasing lifespan. However, high incidence and early onset of aging-related conditions such as frailty, pose as a new threat to this population. OBJECTIVES: We aimed to characterize frailty by comparing health domains consisting of psychosocial, functional and physical deficits between frail PLWH and matched uninfected controls; identify associated risk factors and the impact on negative health outcomes including mortality risk score, quality of life, healthcare utilization, functional disability and history of falls among virally suppressed PLWH. DESIGN: Cross-sectional study. SETTING: Infectious disease clinic in a tertiary institution. PARTICIPANTS: Individuals aged >25 years, on ART >12 months, not pregnant and without acute illness; multi-ethnic, Asian. MEASUREMENTS: Frailty instruments included Frailty phenotype (FP), FRAIL scale (FS) and Frailty index (FI). FI health deficits were categorized into health domains (psychosocial, functional and physical) and used as standard comparator to characterize frailty. Health domains of frail PLWH were compared with frail matched, uninfected controls. Regression analyses were applied to explore associated risk factors and health-related frailty outcomes. RESULTS: We recruited 336 PLWH. Majority were male (83%), Chinese (71%) with CD4+ count 561 (397-738) cells/µl. Frailty prevalence among PLWH were 7% (FP); 16% (FS) and 22% (FI). Proportions of psychosocial, functional, and physical domains were similarly distributed among frail PLWH measured by different frailty instruments. When compared with matched controls, psychosocial dominance was significant among the PLWH, but not in functional and physical domains. Identified frailty risk factors included poor nutritional status, higher CD4+ count nadir, depression, metabolic syndrome, higher highly sensitive C-reactive protein (hsCRP) and history of AIDS-defining illness (ADI). Frailty influenced the risk for negative health outcomes including increased mortality risk scores, poor quality of life (QOL), frequent healthcare utilization and increased functional disability (p<0.05). CONCLUSIONS: This study highlighted the importance of psychosocial influence in the development of frailty among treated PLWH in a multi-ethnic, Asian setting.

Cutaneous vasculitis: a review of aetiology and clinical manifestations in 85 patients in Malaysia.

Cutaneous vasculitis presents with a variety of clinical morphologies and causes significant morbidity. A total of 85 patients with cutaneous vasculitis at Hospital Kuala Lumpur were retrospectively reviewed. Palpable purpura was seen in 49.4% and frequently involved the lower limbs (50.6%). Identifiable causes include drugs (28.2%), infections (20.0%) and connective tissue disorders (16.5%). Non steroidal antiinflammatory were the commonest group of drugs responsible for 25% of cases while B-haemolytic streptococci was the leading infectious cause (64.7%).

Immunomodulating effect of inositol hexaphosphate against Aeromonas hydrophila-endotoxin.

The present study was carried out to evaluate the effect of inositol hexaphosphate (IP6) administration on endotoxemia as an example of the systemic inflammatory response. Mice were divided into three groups as follows: First group, remained as a naive group injected intraperitoneally (i.p.) with PBS (pH 7.4; 0.2 ml/mice) at intervals parallel to the treated groups. The second group was injected i.p. with the lipopolysaccharide (LPS) of Aeromonas hydrophila once a week for four weeks at a dose of LPS suspension: 20 mg/kg mice/week. The third group was injected with the same LPS dose and synergistically intubated with IP6 three times a week for four weeks at a total dose of 4 0mg/kg. At different experimental periods (1, 2, 3 and 4 weeks), six animals from each group were sacrificed under mild diethyl ether anesthesia. Blood and sera were taken for the estimation of phagocytic activity, electrophoretic pattern of proteins and immunoglobulin levels. Also, a slice of liver was homogenized to estimate the respiratory burst enzymes activities and nitric acid synthesis. Histopathological changes of hepatic tissues were investigated. In the LPS-treated group, marked increase in the phagocytic activities and nitric oxide synthesis, and a decrease in hepatocyte catalase, total peroxidase and superoxide dismutase activities were observed. The histopathological features revealed a degeneration and highly mitotic division within the hepatic nuclei in addition to some karyomegaly and nuclear pyknosis. During the treatment period, liver sections of the LPS+IP6 group showed somewhat regenerative features. Reduction in the toxicity of free radicals by IP6 was observed and the IP6 effect seemed to be responsible for the observed ameliorative influence.

Effects of chronic deoxynivalenol exposure on p53 heterozygous and p53 homozygous mice.

Deoxynivalenol (DON) is a secondary metabolite associated with Fusarium species pathogenic to important food crops. A two-year feeding study reported that DON was non-carcinogenic in B6C3F1 mice. The present study was conducted to further characterize the chronic effects of DON by exposing cancer-prone transgenic p53 heterozygous (p53+/-) male mice and p53 homozygous (p53+/+) male mice to 0, 1, 5, or 10 mg DON/kg in diet for 26 weeks. Gross and microscopic organ-specific neoplastic and non-neoplastic changes and expression profiles of key hepatic and renal genes were assessed. Few toxicologic differences between p53+/+ and p53+/- mice were observed, and no tumours were observed due to DON. The results indicated that DON was non-carcinogenic and that reduced expression of the p53 gene did not play a key role in responses to DON toxicity. The lack of inflammatory and proliferative lesions in mice may be attributed to the anorectic effects of DON, which resulted in dose-dependent reductions in body weight in p53+/+ and p53+/- mice. Hepatic and renal gene expression analyses confirmed that chronic exposure to DON was noninflammatory. The effects of 26-week DON exposure on p53+/+ and p53+/-mice were consistent with those previously seen in B6C3F1 mice exposed to DON for two years.

Synthesis of cholesterol modified cationic lipids for liposomal drug delivery of antisense oligonucleotides.

The paper describes a novel synthesis of cholest-5-en-3 beta-yl-6-aminohexyl ether (AH-Chol). AH-Chol was used to prepare positively charged liposomes. The liposomes consisted of phospholipon 90H and the cationic cholesterol derivative in an equimolar ratio. Liposome preparation was achieved by membrane homogenization after rehydration of a dry lipid film. Oligonucleotides (ODN) were adsorbed to the cationic liposomes very efficiently. At an ODN/liposome ratio of 1:5 (10:50 micrograms/ml) 84.2 +/- 5.4% of the ODNs were bound to the liposomal membrane. Within the range of 1:40 and 1:100 charge neutralization occurred and the liposome dispersion showed an increase in particle size due to aggregation. Below or above this range of charge neutralization the ODN loaded liposome preparation was physically stable, no sedimentation, increase of vesicle size or vesicle aggregation occurred.

Significance of immunohistochemical c-ErbB-2 product localisation pattern for prognosis in human breast cancer.

Breast cancer is an increasingly important cause of illness and death among women. In recent years several novel prognostic determinants of breast cancer have been identified, including c-ErbB-2. In this study, expression of c-ErbB-2 in breast carcinoma was correlated with axillary lymph node metastases and disease outcome. The expression of c-ErbB-2 oncoprotein was analysed in 315 tumor specimens of infiltrating ductal carcinoma of breast. They were categorized according to the modified Bloom and Richardson criteria into three histological grades. These patients also had axillary lymph nodes sampling. The expression of c-ErbB-2 oncoprotein was analysed immunohistochemically. Over expression of c-ErbB-2 were observed in 39.36% tumors. Axillary lymph node metastasis had significant correlation with intensified positivity of c-ErbB-2. C-ErbB-2 positive patients did show resistance to chemotherapy when compared for recurrence and distant metastases following surgery (p< 0.05). At a median follow-up of 48 months in c-ErbB-2 positive patients, the overall survival was 3.0 years and disease free survival was 2.5 years. c-ErbB-2 negative tumor patients showed a far better survival. In this group the overall survival was 4.44 years and the disease free survival was 3.78 years. These findings reinforce the view that c-ErbB-2 immunohistochemical detection is of help in detecting a subgroup of breast carcinoma patients who are at high risk. This may also be of particular relevance in decisions regarding adjuvant chemotherapy to these patients.

Evaluation and reinforcement of the correlation of S-phase fraction with nodal metastases, tumor grade and tumor size in breast carcinoma patients in Pakistan.

The aim of this study was to evaluate the S-phase fraction (SPF) of tumors in breast cancer patients in Pakistan. Its association with the traditional morphological prognostic markers, i.e., axillary lymph node metastasis, tumor size and grade, was also studied. Flow cytometry was used to estimate SPF on breast cancer tissues from 166 patients reported at the Aga Khan University Hospital between the years 1997 and 2000. Univariate analysis was done to find any association between SPF and the aforesaid variables. For the ease of analysis, the cases were subdivided into two categories depending on the SPF value, i.e., <10% (low-risk group) and > or = 10% (high-risk group). The mean and median SPF values were 21.45% and 20.035%, respectively, with a range of 3.26% to 54.30%. Twenty-six (15.66%) of the cases had SPF <10%, 57 (34.34%) had SPF from 10%-20%, and 83 (50%) had SPF >20%. A significant correlation between SPF and nodal metastasis was observed (p = 0.0111), but not between SPF and the number of lymph nodes involved when metastatic cases were subdivided into <4 lymph node-positive cases and > or =4 lymph node-positive cases. Significant correlations were also found between SPF and tumor grade (p = 0.0244), as well as between SPF and tumor size (p = 0.048). In conclusion, DNA flow cytometric analysis of SPF carried out in our laboratory could reasonably predict the chances of lymph node metastasis, tumor grade and size in breast cancer patients, thus proving to be an important prognostic marker in the Pakistani setting. This requires further investigations regarding the survival of patients so as to evaluate its capacity of predicting the outcome of Pakistani patients.

Supraventricular arrhythmia: a complication of 5-fluorouracil therapy.

5-Fluorouracil is an S-phase-specific, synthetic pyrimidine antimetabolite, which is used as a cytostatic agent for a variety of malignant lesions, either singly or in multidrug regimens. Gastrointestinal toxicity and myelosuppression are the most common adverse reactions, but, of late, clinical cardiotoxicity has been reported in both prospective and retrospective studies. We present our experience of clinical cardiotoxicity in five patients.

The influence of casting solvent composition on structure and permeability of acrylic-methacrylic ester copolymer films.

Decrease in the solvation of polymer by inclusion of ethanol in the acetone casting solution resulted in greater permeability to urea of cast acrylate-methacrylate film. The greater permeability was accompanied particularly by a decrease in pore size and increase in pore number, despite the absence of change in pore area. A decrease in cohesiveness in the film was suggested by the decrease in tortuosity of the pores as seen by scanning electron microscopy; also, water uptake was increased when the film was cast from an ethanolic solution. The results support the view that, in practice, the composition of the solvent, by its effect on microstructure, can affect the function of the film cast from it.

Profile of Kangari cancer: a prospecive study.

Kangari, an earthenware brazier containing burning charcoal, is used for personal warming during the winter months by most of the people in Kashmir valley. Kangari is usually held close to the lower abdomen or in between the legs while squatting on the ground. The use of kangari is associated with development of skin cancer at the sites of prolonged exposure. In the present study 43 patients with Kangari cancer were analyzed prospectively with the objective of understanding their clinico-pathological profile and the response to various treatment modalities. Variables such as age, gender, grade, site of lesion etc., were evaluated for their prognostic significance.

A syndrome of short stature, mental retardation, facial dysmorphism, short webbed neck, skin changes and congenital heart disease.

We report four sibs from a consanguineous Arab Bedouin family with short stature, mental retardation, a peculiar face, a short webbed neck, skin changes and congenital heart defects. We suggest that the constellation of abnormalities in these children could represent a newly recognized syndrome.

Synthesis and antimicrobial activities of some new 2-substituted benzoxazole/benzothiazole derivatives.

In the search for new antimicrobial compounds, several new sulfur bearing heterobicyclic moieties (4-11) have been synthesized by acylation and alkylation of acetamide, thioacetamide and semicarbazide derivatives. The structure of the products was deduced from elemental analyses as well as spectral data (IR, 1H NMR and MS). Significant antimicrobial activities were obtained for all new compounds especially against Fusarium oxysporum.

Flow cytometric analysis of childhood leukemias.

OBJECTIVE: To collect demographic data for childhood (less than 15 years) leukemias in Karachi, describe the accuracy of the cell surface markers routinely used in the flow cytometric analysis of leukemic cells and arrive at an ideal panel of antibodies for analyzing leukemic samples. MATERIALS AND METHODS: Data from 62 consecutive cases of childhood leukemias referred to the Department of Pathology, Aga Khan University Hospital, (AKUH) between January 1995 and December 1998 was analyzed using Epi Info Version 6. Flow cytometry on all samples was performed using standard protocols. RESULTS: The mean age of patients was 8.2 years and 49 (79%) were males. Fifty (81%) had acute lymphoblastic leukemias of which 50% were CD10 positive and 24% CD10 negative Pre-B cell leukemias. Among all Pre B cell All 98% were positive for CD19, 96% for CD22, 89% for HLA-DR and 67% for CD10. Of the 10 AML cases, 100% were positive for CD33, 90% for CD13, 80% for CD19 and 70% for HLA-DR. CONCLUSION: The mean age in this study population was significantly higher and percentage of CD10 positive Pre-B All is lower than that in the West. Both these factors might be responsible for the poorer prognosis of these patients. It is not possible to specify a minimum or maximum panel of antibodies that should be used for phenotyping all cases of childhood leukemias. A certain degree or redundancy is essential in any panel of antibodies used for flow cytometry of leukemias.

p53 and PCNA expression in benign, atypical and malignant meningiomas.

OBJECTIVE: Alterations: p53 genes are turning out to be the most common genetic alterations in human cancers. Due to long half-life of mutated p53, its detection is possible by immunohistochemistry. Proliferating cell nuclear antigen (PCNA) is expressed by dividing cells, hence has been shown to correlate with prognosis. We have used monoclonal antibodies protein DO-7 (p53) and PC10 (PCNA) to see whether their expression correlates with histological grading in meningethelial tumour. MATERIAL AND METHODS: Twenty nine meningiomas (20 benign, 7 atypical and 2 malignant) were selected from the records of our laboratory. p53 and PCNA expression was sought by immunohistochemistry using Peroxidase Anti Peroxidase (PAP) technique. RESULTS: Four benign and 2 atypical meningiomas showed weak staining for p53. Both malignant meningiomas showed strong positivity for p53. Six benign meningiomas had less than 5% PCNA positivity, one 10% positivity and three showed 20% positivity. PCNA positivity ranged for 10-80% in atypical meningiomas. In two malignant meningiomas PCNA positivity was 70% and 90%. CONCLUSION: It is worthwhile to include p53 and PCNA expression along with histologic assessment in predicting outcome of meningiomas. A larger series with complete follow-up is essential in assessing value of these markers which unfortunately remains a dream in our country.