Molecular mechanisms of extracellular-ATP-mediated colorectal cancer progression: Implication of purinergic receptors-mediated nucleocytoplasmic shuttling of HuR.

One of the leading causes of cancer-related deaths worldwide is colorectal cancer (CRC). Extracellular ATP (e-ATP) and purinergic receptors (P2R) play a central role in CRC proliferation and progression. Human antigen R (HuR) is becoming more and more understood to be essential for the expression of genes linked to cancer. The current study demonstrates that ATP can mediate CRC (Caco-2 cells) progression via induction of HuR nucleocytoplasmic shuttling and subsequent expression of cancer-related genes, a consequence mostly mediated via the P2R receptor. It was also noted that suppression of HuR activity by using dihydrotanshinone I (DHTS) prevents cancer-related gene expression and subsequent CRC (Caco-2 cells) progression induced by ATP. The expression of cyclin A2/cyclin-dependent kinase 2 (CDK2), Bcl-2, ProT-α, hypoxia-inducible factor1-α (HIF1-α), vascular endothelial growth factor A (VEGF-A), transforming growth factor-ß (TGF-ß) and matrix metallopeptidase 9 (MMP-9) induced by ATP were highly reduced in the presence of either PPADS (non-selective P2R antagonist) or DHTS. In addition, e-ATP-induced Caco-2 cell proliferation as well as cell survival were highly reduced in the presence of either PPADS or DHTS or selective CDK-2 inhibitor (Roscovitine) or selective Bcl-2 inhibitor (ABT-263). Furthermore, it was found that MMP-9 is critical for Caco-2 cells migration induced by e-ATP as demonstrated by a clear reduction in cells migration in the presence of a selective MMP-9 inhibitor (Marimastat). Collectively, these data demonstrate that ATP through P2R activation can induce HuR nucleocytoplasmic shuttling that could be translated into an increase in cancer-related genes expression and subsequent, cell proliferation and progression.

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.

Perceived barriers to breast cancer screening among Saudi women at primary care setting.

INTRODUCTION: Screening for breast cancer (BC) is of low rate in Saudi Arabia; although it is provided in the country free of charge to the population. This cross-sectional study aimed at investigating the perceived barriers towards BC screening in Al Hassa, Saudi Arabia.Participants and methods. A total of 816 adult Saudi women aged ≥ 30 years attending for routine primary health services or accompanying patients at the selected primary health care centers (PHCs) were randomly selected from 12 PHCs (8 urban and four rural) using multi-stage sampling method. Participants were invited to personal interview using semi-structured data collection instrument including inquiries about socio-demographics, reproductive history, previous histories of diagnosed breast lesions and breast cancer. The perceived individual barriers towards screening, their attitudes, the reasons for not attending previously held screening campaigns in Al Hassa, were also included. RESULTS: Low utilization of BC screening being significantly positively associated with woman's age, higher educational status, higher family income, using hormonal contraception and positive history of previous breast as shown by the results of the logistic regression model. Exploratory factor analysis showed that personal fears (especially fear of doctors/examiners, fear of hospitals and health facilities and fear of consequences/results) were the major factors that hinder women from utilizing the free of charge BC screening with high loading eigenvalue of 3.335, explaining 30.4% of the barriers. CONCLUSIONS: Educational interventions aim at improving breast cancer knowledge and addressing barriers should be incorporated as core component of the screening program in Saudi Arabia.

Effect of dietary counseling on preventing excessive weight gain during pregnancy.

OBJECTIVES: Excessive gestational weight gain (EGWG) is associated with short- and long-term health problems among mothers and their offspring. The aim of this study was to assess the effect of dietary counseling on EGWG. STUDY DESIGN: Randomized controlled intervention trial. METHODS: The study was conducted at the antenatal care (ANC) clinic, Center for Social and Preventive Medicine, Pediatrics Hospital, Cairo University, during the period from July 2015 to April 2016. A total of 200 primigravidae aged between 20 and 30 years were included and randomized into intervention and control groups. Based on data analysis from phase 1, nutritional practices of the studied participants were identified and tailored nutrition counseling sessions designed. Follow-up of the studied participants throughout ANC visits was done to enforce the healthy dietary intake and encourage weight gain according to the recommendations to avoid EGWG. RESULTS: The intervention resulted in a significantly higher proportion of women in the intervention group who gained gestational weight within the Institute of Medicine recommendations compared to women in the control group (42.7% vs 13.9%, respectively) (P-value <0.001); this was detected between the 25th and 35th weeks of gestation. Dietary counseling had significantly improved the frequency of consumption of different food items and knowledge, attitude, and practice mean scores of the intervention group in comparison with the control group (P-value <0.001). CONCLUSIONS: The results of the present study demonstrated that dietary counseling given to pregnant women reduced the proportion of EGWG and improved dietary practices. There is a strong need for effective intervention strategies targeting EGWG to prevent adverse pregnancy outcomes.

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.

Structural and functional effects of early exposure to 4-nonylphenol on gonadal development of Nile tilapia (Oreochromis niloticus): a-histological alterations in ovaries.

The present study investigated the gonads alterations of mature female Oreochromis niloticus caused by different concentrations (0, 40, 60, 100 µg/L) of 4-nonylphenol (NP) and also its effect on steroidogenesis. A tendency for a dose-dependent reduction in the gonadosomatic index, fecundity and oocytes diameter in the NP-exposed groups was observed. Histological examination revealed that NP impairs gonadal growth clearly shown in the oocyte development and differentiation. Also, the estrogenic activity of such NP was identified through 11-ketotestosterone, 17b-estradiol and vitellogenin which were affected significantly by the applied concentrations. The study indicates that NP has estrogenic potency-induced marked alteration in the sexual development including gonadal maturation, spawning time and egg production.

Structural and functional effects of early exposure to 4-nonylphenol on gonadal development of Nile tilapia (Oreochromis niloticus): b-histological alterations in testes.

The present study investigates the effects of different doses (0, 40, 60, 100 µg/L) of 4-nonylphenol (NP) on testis histology and sperms motility of mature Oreochromis niloticus. A tendency for a dose-dependent reduction in the gonado-somatic index (GSI) in the NP-exposed groups was observed. Histological examination revealed that high doses of NP (60 and 100 µg/L) cause hyperplasia of interlobular connective tissue components coincide with decrease in the size of seminiferous lobules and amount of lumen spermatozoa in comparison with the control and low-dose (40 µg/L)-exposed groups. In addition, within the seminiferous lobules, rupture of spermatogenic cysts, vacuolation and necrosis of primary spermatocytes due to damage of sertoli cells were clearly observed and lead to decrease in advanced spermatogenic cells. The highest dose caused significant decrease in the GSI associated with appearance of testis-ova (intersex) state. The percentage of abnormal sperms increased with increasing the dose of NP reaching to 96 % for those subjected to 100 µg NP/L, while it was only 11 % for the control group specimens. The study indicates that NP has estrogenic potency induced marked alteration in the sexual development including inhibition in testicular growth and deformation in the sperms.

Study of the effect of dietary counselling on the improvement of end-stage renal disease patients.

Proper nutrition may help to reverse the wasting syndrome in dialysis patients with kidney disease on dialysis. This intervention study aimed to identify malnutrition problems and assess the effect of dietary counselling on improvement of health status of end-stage renal disease patients subjected to haemodialysis. Pre-intervention data were collected from 41 patients attending El Haram dialysis centre, Giza, Egypt; 97.5% of the patients were considered mildly to moderately malnourished and multiple malnutrition problems were detected (protein-energy malnutrition, hypocalcaemia, and anaemia and hyperphosphataemia). Nutritional counselling sessions resulted in statistically significant post-intervention improvements in Karnofsky performance scale scores, malnutrition inflammation scores and nutritional knowledge of the patients. Providing one-to-one nutrition counselling could be linked to improvements in the patients' nutritional knowledge and practices and to their health status and performance in activities of daily life.

Study of the effect of dietary counselling on the improvement of end-stage renal disease patients

Proper nutrition may help to reverse the wasting syndrome in dialysis patients with kidney disease on dialysis. This intervention study aimed to identify malnutrition problems and assess the effect of dietary counselling on improvement of health status of end-stage renal disease patients subjected to haemodialysis. Pre-intervention data were collected from 41 patients attending El Haram dialysis centre, Giza, Egypt; 97.5% of the patients were considered mildly to moderately malnourished and multiple malnutrition problems were detected [protein-energy malnutrition, hypocalcaemia, and anaemia and hyperphosphataemia]. Nutritional counselling sessions resulted in statistically significant post-intervention improvements in Karnofsky performance scale scores, malnutrition inflammation scores and nutritional knowledge of the patients. Providing one-to-one nutrition counselling could be linked to improvements in the patients' nutritional knowledge and practices and to their health status and performance in activities of daily life

HCV infection among Saudi population: high prevalence of genotype 4 and increased viral clearance rate.

HCV is a major etiological agent of liver disease with a high rate of chronic evolution. The virus possesses 6 genotypes with many subtypes. The rate of spontaneous clearance among HCV infected individuals denotes a genetic determinant factor. The current study was designed in order to estimate the rate of HCV infection and ratio of virus clearance among a group of infected patients in Saudi Arabia from 2008 to 2011. It was additionally designed to determine the genotypes of the HCV in persistently infected patients. HCV seroprevalence was conducted on a total of 15,323 individuals. Seropositive individuals were tested by Cobas AmpliPrep/Cobas TaqMan HCV assay to determine the ratio of persistently infected patients to those who showed spontaneous viral clearance. HCV genotyping on random samples from persistently infected patients were conducted based on the differences in the 5'untranslated region (5'UTR). Anti-HCV antibodies were detected in 7.3% of the totally examined sera. A high percentage of the HCV infected individuals experienced virus clearance (48.4%). HCV genotyping revealed the presence of genotypes 1 and 4, the latter represented 97.6% of the tested strains. Evidences of the widespread of the HCV genotype 4 and a high rate of HCV virus clearance were found in Saudi Arabia.

Immobilization of low and intermediate level of organic radioactive wastes in cement matrices.

The adequacy of cement-clay composite, for solidification/stabilization of organic radioactive spent liquid scintillator wastes and its resistance to frost attack were determined by a freezing/thawing (F/T) test. Frost resistance is assessed for the candidate cement-clay composite after 75 cycles of freezing and thawing by evaluating their mass durability index, compressive strength, apparent porosity, volume of open pores, water absorption, and bulk density. Infrared (IR), X-ray diffraction (XRD), differential thermal analysis (DTA), thermal gravimetric analysis (TGA) and scanning electron microscopy (SEM) were performed for the final waste form (FWF) before and after the F/T treatment to follow the changes that may take place in its microstructure during the hydration regime. The results were obtained indicate that the candidate composite exhibits acceptable resistance to freeze/thaw treatment and has adequate suitability to solidify and stabilize organic radioactive spent liquid scintillator wastes even at very exaggerating conditions (-50°C and +60°C).

Molecular evolution of the six internal genes of H5N1 equine influenza A virus.

Phylogenetic and evolutionary patterns of the six internal genes of an equine H5N1 influenza A virus isolated in Egypt on 2009 were analyzed using direct sequencing. All of the internal genes of the equine H5N1 strain showed a genetic pattern potentially related to Eurasian lineages. Variable dendrogram topologies revealed an absence of reassortment in the equine strain while confirming its close relatedness to other Egyptian H5N1 strains from human and avian species. The equine strain is characterized by a variety of amino acid substitutions in six internal proteins compared to the available Egyptian H5N1 strains. Interestingly, the equine strain displayed amino acids in the PB2, PA, M2 and NS2 proteins that are unique among the available H5N1 sequences in the flu database, and their potential effect on virulence needs to be further investigated.

Synthesis of new 2-naphthyl ethers and their protective activities against DNA damage induced by bleomycin-iron.

The reaction of 2-naphthaloxyacetic acid with thiosemicarbazide in the presence of phosphoryl chloride, followed by treatment with phenacylbromides, led to the formation of imidazo[2,1-b][1,3,4]thiadiazoles 3a-c. 2-(Naphthalen-3-yloxy)acetohydrazide 4 on treatment with ethyl 2-(2-arylhydrazono)-3-oxobutanoates (5a-c), 2-methoxymethylene)malononitrile, or ethyl 2-cyano-3,3-bis(methylthio)acrylate led to the formation of substituted pyrazoles 6-8. The reaction of the hydrazide 4 with hydrazonoyl chlorides 9a-c and 1,2,4,5-benzene tetracarboxylic-1,2:4,5-dianhydride produced bis-diazo compounds 10a-c and dimide 11 respectively. All new compounds were tested for their protective activity against DNA damage induced by bleomycin-iron complex. Compound 2 showed the greatest protection against DNA damage, thus diminishing chromogen formation between the damaged DNA and thiobarbituric acid.

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.

Modulation of macrophage functions by sheeppox virus provides clues to understand interaction of the virus with host immune system.

BACKGROUND: Poxviruses encode a range of immunomodulatory genes to subvert or evade the challenges posed by the innate and adaptive immune responses. However, the inactivated poxviruses possessed immunostimulating capacity and were used as a prophylactic or metaphylactic application that efficiently reduced susceptibility to infectious diseases in different species. This fact is intensively studied in different genera of poxviruses. However, little is known about the basic mechanisms adopted by sheeppox virus (SPPV). SPPV causes an acute disease of sheep that recently, has been observed to reinfect its host in spite of vaccination. RESULTS: By injecting inactivated or attenuated sheeppox virus SPPV vaccine in adult male Swiss mice, SPPV was found to reduce macrophages' functions in a local event that occurs at the site of application 12 h after vaccine administration as indicated by increased level of IL-10 and decreased level of SOD from cultured peritoneal macrophages. In contrast increased levels of IL-12, and SOD activity from cultured splenic macrophages, lymphocyte response to PHA-P, and in-vivo response to T-dependant Ag were detected. These effects were observed in both attenuated and inactivated SPPV, but more prominent in attenuated one. CONCLUSION: The results of this study help to elucidate, the phenomenon of existence natural SPPV infections in sheep instead of vaccination and the basic mechanisms responsible for the immunostimulating capacity of sheeppox virus. Locally, SPPV shows evidence for an immune escape mechanism that alleviates the host's immune response. Later and systemically, the virus protects the host from any fatal consequences of the immune system suppression.

Evaluation of the effect of a plant alkaloid (berberine derived from Berberis aristata) on Trichomonas vaginalis in vitro.

Berberine is a quaternary alkaloid derived from the plant Berberis aristata having antibacterial, antiamoebic, antifungal, antihelminthic, leishmanicidal and tuberculostatic properties. The effect of berberine sulphate salt on the growth of Trichomonas vaginalis in vitro was compared to the efficacy of metronidazole as a reference drug. Results showed that berberine sulphate was comparable to metronidazole as regards potency with the advantage of being more safe and possible replacement in metronidazole resistant cases.

Assessment of the role of soluble intracellular adhesion molecule-1 (sICAM-1) in the pathogenesis and as a marker of disease severity in different stages of human schistosomiasis and toxoplasmosis.

The role of soluble intracellular adhesion molecule 1 (sICAM-1) in the pathogenesis and its relevance to disease severity was assessed in different stages of human infection with S. mansoni, S. haematobium and T. gondii. The levels of sICAM-1, obtained in the current study correlated with disease severity, degree of cell destruction and type of immune response. Highest sICAM-1 levels were observed in only two groups (hepatosplenic schistosomiasis and patients with mixed active S. haematobium and T. gondii infections), while in the other two groups (separate cases of active S. haematobium infection or latent T. gondii infection) showed no significant rise in sICAM-1 levels.

Experimental trials of an artemisinin derivative in treatment of Schistosoma mansoni infected mice.

Dihyroartemesinin, an artemesinin derivative, was used for treatment of mice experimentally infected with S. mansoni. A significant decrease in total worm count was recorded when mice were treated during early and late stages of infection. Histopathological examination of liver sections showed minimal pathological changes with early treatment and enhanced fibrosis of granulomas with late treatment. The results showed that dihydroartemesinin had a promising effect on S. mansoni infected mice almost comparable to that of praziquantel.

Evaluation of aminoalkylmethacrylate nanoparticles as colloidal drug carrier systems. Part II: characterization of antisense oligonucleotides loaded copolymer nanoparticles.

Aminoalkylmethacrylate methylmethacrylate copolymer nanoparticles were evaluated for their use as potential drug carrier systems. Their cytotoxicity, as well as the loading of antisense oligonucleotides that were employed as anionic model drugs depended on the substitution of the basic aminoalkyl copolymer. Toxic influences on the integrity of cell membranes depended on aminoalkyl groups located on the particle surfaces. Toxicity was observed either by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays using African green monkey kidney (AGMK) cells or by a hemolysis test, where the efflux of haemoglobin from disrupted erythrocytes was measured. The cytotoxic effects were increased by the elongation of the N-alkyl chain by four additional methylene groups. Lipophilic polymethylmethacrylate (PMMA) homopolymer nanoparticles showed a negative surface charge and, therefore, were not suitable for the adsorption of anionic drugs. The surface charge was changed to positive values by the incorporation of basic monomers. Consequently, the loading efficacy was increased by raising the basic copolymer portion. Additionally, a pH-dependent loading behaviour of oligonucleotides was observed. Substitution of the amino nitrogen protons by methyl groups led to a decreased oligonucleotide loading and to a reduced cytotoxicity. Nanoparticles with permanent positively charged quarternary ammonium groups showed a high pH-independent loading efficacy, but also possessed a high cytotoxic potential. In this study, cationic copolymer nanoparticles containing 30% (w/w) methylaminoethyl-methacrylate (MMAEMC) were found to be optimal with regard to biocompatibility and carrier properties for hydrophilic anionic antisense oligonucleotides. A significant portion of adsorbed oligonucleotides were protected from enzymatic degradation. The cellular uptake of oligonucleotides into Vero cells was significantly enhanced by this methylaminoethyl-methacrylate derivative.