Molecular and cellular study of liver diseases: focusing on the biology of toll-like receptors in liver tumours

Background. Liver cancer is the 3rd commonest cause of cancer death worldwide. Understanding the mechanisms of hepatocarcinogenesis is vital for developing more effective treatments. Chronic liver disease is a predisposing factor for development of hepatocellular carcinoma (HCC) and increased translocation of gut bacteria is believed to exacerbate this inflammatory condition. Toll-like receptors (TLRs) play a crucial role in immunity against microbial pathogens and recent evidence suggests they may be important in pathogenesis of chronic liver disease. The aim of this thesis was to explore the role of TLRs in the pathogenesis of HCC. Materials & Methods: Tissue microarrays obtained from patients with cirrhosis, viral hepatitis and HCC were stained with for TLR4, TLR7 and TLR9 and the data were validated in actual patient samples. The role of gut translocation was explored in an animal model of HCC using diethylnitrosamine (DEN) and nitrosomorpholine (NMOR) after treatment with Norfloxacin. Proliferation of HCC cell lines were studied after stimulation (Imiquimod and CpG-ODN) and inhibition of TLR7 and TLR9 (IRS 954) and chloroquine. The effect of these interventions was confirmed in the DEN and NMOR and, a xenograft model. The studies were extended to determine their effect in cholangiocarcinoma. Results: TLR7 and TLR9 but not TLR4 were up regulated in HCC tissue and gut decontamination with Norfloxacin did not prevent HCC development but reduced liver fibrosis. TLR7 stimulation increased cell proliferation of HuH7 cells significantly and inhibition of TLR7 and TLR9 using IRS or chloroquine resulted in significant inhibition. TLR7 and TLR9 inhibition using IRS 954 and chloroquine reduced tumour growth in xenograft models and, chloroquine also decreased liver fibrosis and tumour growth in the DEN and NMOR model. These beneficial effects were also observed in cholangiocarcinoma. Conclusion: In conclusion, these data suggest that inhibiting TLR7 and TLR9 and, using chloroquine could be potential novel therapeutic strategies for the prevention and the progression of primary liver cancers in susceptible patients

Room-temperature synthesis of zinc oxide nanoparticles in different media and their application in cyanide photodegradation

Cyanide is an extreme hazard and extensively found in the wastes of refinery, coke plant, and metal plating industries. A simple, fast, cost-effective, room-temperature wet chemical route, based on cyclohexylamine, for synthesizing zinc oxide nanoparticles in aqueous and enthanolic media was established and tested for the photodegradation of cyanide ions. Particles of polyhedra morphology were obtained for zinc oxide, prepared in ethanol (ZnO(E)), while spherical and some chunky particles were observed for zinc oxide, prepared in water (ZnO(W)). The morphology was crucial in enhancing the cyanide ion photocatalytic degradation efficiency of ZnO(E) by a factor of 1.5 in comparison to the efficiency of ZnO(W) at an equivalent concentration of 0.02 wt.% ZnO. Increasing the concentration wt.% of ZnO(E) from 0.01 to 0.09 led to an increase in the photocatalytic degradation efficiency from 85% to almost 100% after 180 min and a doubling of the first-order rate constant (k)

Promoter Hypermethylation and Suppression of Glutathione Peroxidase 3 Are Associated with Inflammatory Breast Carcinogenesis

Reactive oxygen species (ROS) play a crucial role in breast cancer initiation, promotion, and progression. Inhibition of antioxidant enzymes that remove ROS was found to accelerate cancer growth. Studies showed that inhibition of glutathione peroxidase-3 (GPX3) was associated with cancer progression. Although the role of GPX3 has been studied in different cancer types, its role in breast cancer and its epigenetic regulation have not yet been investigated. The aim of the present study was to investigate GPX3 expression and epigenetic regulation in carcinoma tissues of breast cancer patients’ in comparison to normal breast tissues. Furthermore, we compared GPX3 level of expression and methylation status in aggressive phenotype inflammatory breast cancer (IBC) versus non-IBC invasive ductal carcinoma (IDC). We found that GPX3 mRNA and protein expression levels were downregulated in the carcinoma tissues of IBC compared to non-IBC. However, we did not detect significant correlation between GPX3 and patients’ clinical-pathological prosperities. Promoter hypermethylation of GPX3 gene was detected in carcinoma tissues not normal breast tissues. In addition, IBC carcinoma tissues showed a significant increase in the promoter hypermethylation of GPX3 gene compared to non-IBC. Our results propose that downregulation of GPX3 in IBC may play a role in the disease progression

Influence of self-efficacy management on adherence to self-care activities and treatment outcome among diabetes mellitus type 2 Sudanese patients

Background: High level of self-efficacy and adherence to self-care activities have a positive impact on the achievement of glycemic goal among diabetic patients. In Sudan, there is a gap in knowledge related to self-efficacy management and its influence on adherence to self-care activities and overall disease control. Objective: To identify the influence of management self-efficacy on adherence to self-care activities and treatment outcome among Sudanese patients with type 2 diabetes mellitus. Methods: A cross-sectional study was conducted at two health care facilities in Sudan from April to May 2016. Patients with type 2 diabetes mellitus were included. Convenience sampling method was adopted. Diabetes Management Self-Efficacy Scale and the Revised Summary of Diabetes Self-care Activities were used to collect data through a face-to-face interview. Logistic regression analysis was performed. A p value <0.05 was considered to be significant. Data were processed using the software SPPS v 21.0. Results: A total of 392 patients were included. Respondents classified with high level of self- efficacy across all domains were 191 (48.7%). Moreover, high level of education [adjusted OR 0.5 (0.3-0.7), (p=0.001)] and formal health education on diabetes [adjusted OR 2.4 (1.6-3.7), (p<0.001)], were found to be significantly associated with high level of diabetes management self-efficacy. Patients who had high level of self-efficacy to manage nutrition, physical exercise activity and medication were found more adherent to general diet, exercise activity, and medication taking, respectively. Patients with controlled disease were 87(22.2%). The only predictor of diabetes control was diabetes management self-efficacy [OR 2.1(1.3- 3.5), (p=0.002)]. Conclusions: Diabetes management self-efficacy was associated with high level of education and receiving health education. Self-efficacy was significantly associated with adherence to self-care activities and glycemic control. Substantial efforts are still needed to empower the patients with self-efficacy and improving adherence to self-care activities through appropriate interventions

Antiangiogenic properties of Koetjapic acid, a natural triterpene isolated from Sandoricum koetjaoe Merr

Background: Angiogenesis, the formation of new blood vessels, has become an important target in cancer therapy. Angiogenesis plays an important role in tumor growth and metastasis. Koetjapic acid (KA) is a seco-A-ring oleanene triterpene isolated from S. koetjape. The solvent extract of this plant species was shown previously to have strong antiangiogenic activity; however the active ingredient(s) that conferred the biological activity and the mode of action was not established. Given the high concentration of KA in S. koetjape, an attempt has been made in this study to investigate the antiangiogenic properties of KA.Results: Treatment with 10-50 μg/ml KA resulted in dose dependent inhibition of new blood vessels growth in ex vivo rat aortic ring assay. KA was found to be non-cytotoxic against HUVECs with IC40.97 ± 0.37 μg/ml. KA inhibited major angiogenesis process steps, endothelial cell migration and differentiation as well as VEGF expression.Conclusions: The non-cytotoxic compound, KA, may be a potent antiangiogenic agent; its activity may be attributed to inhibition of endothelial cells migration and differentiation as well VEGF suppression

Evaluation of Xpert® MTB/RIF and ustar easyNAT™ TB IAD for diagnosis of tuberculous lymphadenitis of children in Tanzania : a prospective descriptive study

Fine needle aspiration biopsy has become a standard approach for diagnosis of peripheral tuberculous lymphadenitis. The aim of this study was to compare the performance of Xpert MTB/RIF and Ustar EasyNAT TB IAD nucleic acid amplification assays, against acid-fast bacilli microscopy, cytology and mycobacterial culture for the diagnosis of TB lymphadenitis in children from a TB-endemic setting in Tanzania.; Children of 8 weeks to 16 years of age, suspected of having TB lymphadenitis, were recruited at a district hospital in Tanzania. Fine needle aspirates of lymph nodes were analysed using acid-fast bacilli microscopy, liquid TB culture, cytology, Xpert MTB/RIF and EasyNAT. Latent class analysis and comparison against a composite reference standard comprising "culture and/or cytology" was done, to assess the performance of Xpert MTB/RIF and EasyNAT for the diagnosis of TB lymphadenitis.; Seventy-nine children were recruited; 4 were excluded from analysis. Against a composite reference standard of culture and/or cytology, Xpert MTB/RIF and EasyNAT had a sensitivity and specificity of 58 % and 93 %; and 19 % and 100 % respectively. Relative to latent class definitions, cytology had a sensitivity of 100 % and specificity of 94.7 %.; Combining clinical assessment, cytology and Xpert MTB/RIF may allow for a rapid and accurate diagnosis of childhood TB lymphadenitis. Larger diagnostic evaluation studies are recommended to validate these findings and on Xpert MTB/RIF to assess its use as a solitary initial test for TB lymphadenitis in children

Synchronous fluorescence spectrofluorimetric method for the simultaneous determination of metoprolol and felodipine in combined pharmaceutical preparation

A rapid, simple and sensitive synchronous specrtofluorimetric method has been developed for the simultaneous analysis of binary mixture of metoprolol (MTP) and felodipine (FDP). The method is based upon measurement of the synchronous fluorescence intensity of the two drugs at Δλ of 70 nm in aqueous solution. The different experimental parameters affecting the synchronous fluorescence intensities of the two drugs were carefully studied and optimized. The fluorescence intensity-concentration plots were rectilinear over the ranges of 0.5-10 μg/mL and 0.2-2 μg/mL for MTP and FDP, respectively. The limits of detection were 0.11 and 0.02 μg/mL and quantification limits were 0.32 and 0.06 μg/mL for MTP and FDP, respectively. The proposed method was successfully applied for the determination of the two compounds in their commercial tablets and the results obtained were favorably compared to those obtained with a comparison method

Parvovirus B19 infection in Tunisian patients with sickle-cell anemia and acute erythroblastopenia

<p>Abstract</p> <p>Background</p> <p>Human parvovirus B19 is the etiologic agent of erythema infectiosum in children. It is also associated with other clinical manifestations in different target groups. Patients with chronic hemolytic anemia are at high risk of developing acute erythroblastopenia following infection by the virus. They usually become highly viremic and pose an increased risk of virus transmission. Close monitoring of such high risk groups is required for epidemiologic surveillance and disease prevention activities. Here we report a molecular epidemiological study on B19 virus infection in Tunisian patients with chronic hemolytic anemia.</p> <p>Methods</p> <p>This study was conducted on 92 young chronic hemolytic anemia patients who attended the same ward at the National Bone Marrow Transplantation Center of Tunis and 46 controls from a different hospital. Screening for IgM and IgG anti-B19 antibodies was performed using commercially available enzyme immunoassays and B19 DNA was detected by nested PCR in the overlapping <it>VP1/VP2 </it>region. DNA was sequenced using dideoxy-terminator cycle sequencing technology.</p> <p>Results</p> <p>Anti-parvovirus B19 IgG antibodies were detected in 26 of 46 sickle-cell anemia patients, 18 of 46 β-thalassemia and 7 of 46 controls. Anti-parvovirus B19 IgM antibodies were detected only in 4 of the sickle-cell anemia patients: two siblings and two unrelated who presented with acute erythroblastopenia at the time of blood collection for this study and had no history of past transfusion. B19 DNA was detected only in sera of these four patients and the corresponding 288 bp nested DNA amplicons were sequenced. The sequences obtained were all identical and phylogenetic analysis showed that they belonged to a new B19 virus strain of Genotype1.</p> <p>Conclusion</p> <p>A new parvovirus B19 strain of genotype1 was detected in four Tunisian patients with sickle-cell anemia. Virus transmission appeared to be nosocomial and resulted in acute erythroblastopenia in the four patients. The possibility of independent transmission of this B19 variant to the patients is unlikely in light of the present epidemiological data. However this possibility cannot be ruled out because of the low genetic variability of the virus.</p

Induction of protein citrullination and auto-antibodies production in murine exposed to nickel

Abstract Citrullination, or the post-translational deimination of polypeptide-bound arginine, is involved in several pathological processes in the body, including autoimmunity and tumorigenesis. Recent studies have shown that nanomaterials can trigger protein citrullination, which might constitute a common pathogenic link to disease development. Here we demonstrated auto-antibody production in serum of nanomaterials-treated mice. Citrullination-associated phenomena and PAD levels were found to be elevated in nanomaterials -treated cell lines as well as in the spleen, kidneys and lymph nodes of mice, suggesting a systemic response to nanomaterials injection, and validated in human pleural and pericardial malignant mesothelioma (MM) samples. The observed systemic responses in mice exposed to nanomaterials support the evidence linking exposure to environmental factors with the development of autoimmunity responses and reinforces the need for comprehensive safety screening of nanomaterials. Furthermore, these nanomaterials induce pathological processes that mimic those observed in Pleural MM, and therefore require further investigations into their carcinogenicity