Bio-diagnostic performances of microRNAs set related to DNA damage response pathway among hepatitis C virus-associated hepatocellular carcinoma patients.

BACKGROUND: Up to date, a well-defined microRNAs (miRNAs) profile involved in hepatocellular carcinoma (HCC) pathogenesis remains indecisive. Thus, employing miRNAs for HCC diagnosis is demanded for early therapeutic interventions. We aimed to evaluate the usage of miRNAs set related to the SuperPath: miRNAs involved in DNA damage response pathway as effective biomarkers for HCV-related HCC diagnosis. RESULTS: The study enrolled 97 patients with HCV-related HCC, 84 with hepatitis C virus (HCV), 97 with liver cirrhosis (LC), and 84 healthy individuals. Serum miRNA-23a, miRNA-203, miRNA-100-5p, and miRNA-16 were quantified using qRT-PCR experiments, AFP and routine LFTs were estimated via standard techniques. Pathway enrichment analysis along with the construction of miRNAs regulatory network were performed. With respect to healthy individuals, miRNA-203, miRNA-100-5p, and miRNA-16 were significantly downregulated in HCC, HCV, and LC groups, while miRNA-23a showed significant upregulation (p < 0.001). miRNAs exhibited significant correlations with AFP, ALT, AST, and albumin. Also, elevated levels of miRNA-23a were recognized in patients with multiple focal lesions and/or lesion size > 5 cm. Additionally, the diagnostic performance of miRNA-23a expression level at a selected cut-off value of 3.99 overtakes AFP, while expressions of miR-203, miRNA-100-5p, and miRNA-16 represent poor diagnostic outcomes. CONCLUSIONS: Keeping in mind the individual variability and high level of heterogeneity in HCC, our data revealed the diagnostic value of miRNA-23a expression in HCV-related HCC patients. Further extra in silico HCC-specific microRNAs sets are demanded in diagnosis.

Inflammatory and endothelial dysfunction indices among Egyptian females with obesity classes I-III.

BACKGROUND: Obesity is an alarming threat to health in Egypt. More than one in three Egyptians is obese, the highest rate in the world. We aimed to delineate the variability of inflammation and endothelial dysfunction markers among Egyptian females with different obesity classes. METHODS: Out of 130 females, 70 were categorized into three obesity groups: Class I, body mass index (BMI) 30-34.9 kg/m2; Class II, BMI 35-39.9 kg/m2 and Class III BMI ≥ 40 kg/m2, besides 60 control subjects. Anthropometric measurements were recorded and serum levels of tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), interleukin (IL) 6 (IL-6), IL-12, soluble intercellular adhesion molecule 1 (sICAM-1) and soluble vascular adhesion molecule 1 (sVCAM-1) were assessed among participants. RESULTS: In all three classes of obesity, significant increase (P <0.05) in BMI, waist-hip ratio, fat mass and body fat mass % were noted. CRP and sVCAM-1 levels were increased among the three obesity groups. TNF-α levels were increased in class II and III obesity groups. IL-6 and IL-12 levels were elevated in class I and class III groups. While, ICAM-1 levels were increased in class III obesity group. CONCLUSION: Based on individuals' BMI, serum levels of TNF-α, CRP, IL-6, IL-12, sVCAM-1 and sICAM-1 are differentially altered with the progression of obesity. We strongly support the hypothesis that, as the obesity rate is still mounting, a subclinical inflammatory reaction has a role in pathogenesis of obesity and emphasize the elevation of endothelial dysfunction in individuals with obesity.

A novel HCV electrochemical biosensor based on a polyaniline@Ni-MOF nanocomposite.

Hepatitis-C virus ribonucleic acid (HCV-RNA) recognition and quantification based on real-time polymerase chain reaction (RT-PCR) is key to infection control, management, and response to treatment due to its specificity, sensitivity, and quantification capabilities. However, the high cost, time requirements, and need for sophisticated laboratory infrastructure have limited the use of this method in rapid screening, blood banks, and point-of-care testing (POCT). In this work, a novel label-free electrochemical biosensor constructed using a polyaniline@nickel metal-organic framework (Ni-MOF) nanocomposite was developed for direct detection of unamplified HCV nucleic acid. A robust biosensor was fabricated using smooth layer-by-layer deposition of the polyaniline@Ni-MOF nanocomposite, deoxyribonucleic acid (DNA) probe, and bovine serum albumin (BSA) onto a glassy carbon electrode (GCE) and was subsequently monitored real-time via cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The sensitivity and specificity of the newly developed biosensor were specifically examined using the EIS approach. The results revealed that the novel biosensor is highly efficient in quantitative sensing of the HCV target in the presence of nonspecific nucleic acids over the range of 1 fM-100 nM with a detection limit of 0.75 fM (at a S/N ratio of 3). To the best of the authors' knowledge, the proposed biosensor is superior to other MOF platforms. These research findings are expected to have a positive influence on the quantitative detection of HCV RNA and other nucleic acids by offering exceptional accuracy and cost effectiveness, especially in low resource countries. Moreover, this biosensor could be simply adopted for full automation and used in point-of-care testing.

New Biomarkers as Prognostic Factors for Cardiovascular Complications in Type 2 Diabetic Patients.

This study was initiated to explore some novel biomarkers like pro-inflammatory markers (chemerin and visfatin) and anti-inflammatory marker (omentin-1) as prognostic factors for cardiovascular complications in type 2 diabetic patients. Forty diabetic patients without cardiovascular disease, 40 diabetic patients with cardiovascular disease and twenty healthy control counterparts were included in this study. Serum chemerin, omentin-1 and visfatin levels were quantified. Receiver operating characteristic curve analysis was done to identify the cut off value for each marker. The mean serum level of chemerin was 57.65 ± 15.69 ng/l in diabetic patients versus 93.97 ± 26.62 ng/l for the cardio-diabetic ones (P < 0.0001). The mean serum level of omentin-1 was 8.77 ± 1.53 ng/ml in diabetic patients versus 1.76 ± 0.96 ng/ml for the cardio-diabetic ones (P < 0.0001). The mean level of visfatin was 1.44 ± 0.71 ug/l in diabetic patients versus 3.92 ± 3.32 ug/l for the cardio-diabetic ones (P < 0.0001). Chemerin and Visfatin levels were significantly enhanced in the cardio-diabetic patients with increasing C-reactive protein (CRP), triglycerides (TG), fasting blood glucose (FBG), micro-albumin and cholesterol. Omentin-1 level was significantly reduced in the cardio-diabetic patients with increasing CRP, TG, FBG, and cholesterol. It was observed that the area under curve for chemerin, omentin-1and visfatin was 0.877, 0.998 and 0.735, respectively. In conclusion, this study evidences that the measuring serum levels of chemerin, omentin-1 and visfatin may help in the prognosis of cardiovascular complications in type 2 diabetic patients.

Nucleic acids biosensors based on metal-organic framework (MOF): Paving the way to clinical laboratory diagnosis.

Development of ultra-sensitive, high specific and cost-effective nucleic acids (NAs) biosensors is critical for early diagnosis of cancer, genetic diseases and follows up response to treatment. Metal-organic frameworks (MOFs) as sensing materials underwent significant development in recent years due to their unique merits, such as structural diversity, tunable pore scale, large surface area, remarkable adsorption affinities, and good thermal stability. MOFs have shown potential contribution in nucleic acids biosensors research. Herein, a comprehensive overview on NAs biosensors state of the art based on MOFs has been discussed extensively, including different MOFs platforms sensing strategies (fluorescence, electrochemistry, electrochemiluminescence, and colorimetric techniques), their analytical performance and figures of merit in clinical diagnostics, with the future perspective in introducing MOFs in clinical laboratory diagnostics. Moreover, the different MOFs synthesis methods have been highlighted to serve as a guide for the researchers in selecting the appropriate platform that suits their research needs, and applications.

Biochemical and molecular evidences for the antitumor potential of Ginkgo biloba leaves extract in rodents.

Hepatocellular carcinoma (HCC) is one of the deadliest primary cancers, with a 5-year survival rate of 10% or less. This study was undertaken to elucidate the underlying biochemical and molecular mechanisms in favor of N-nitrosodiethylamine-induced hepatocellular carcinoma. Furthermore, the aim of this work was extended to explore the efficacy of Ginkgo biloba leaves extract in deterioration of HCC in rats. In the current study, HCC group experienced significant downregulation of ING-3 gene expression and upregulation of Foxp-1 gene expression in liver. Treatment of HCC groups with Ginkgo biloba leaves extract resulted in upregulation of ING-3 and downregulation of Foxp-1 gene expression in liver. In addition, there was significant increase in serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA) and glypican-3 (GPC-3) levels in HCC group versus the negative control group. In contrast, the groups with HCC subjected to either high or low dose of Ginkgo biloba leaves extract elicited significant reduction (P<0.05) of AFP, CEA and GPC-3 in serum compared to the untreated HCC rats. Besides, histological examination of liver tissue sections of rats in HCC group revealed typical anaplasia. Interestingly, treatment with Ginkgo biloba leaves extract elicited marked improvement in the histological feature of liver tissue in HCC groups. In conclusion, this research indicated that the carcinogenic potency of N-nitrosodiethylamine targeted multiple systems on the cellular and molecular levels. In addition, the results of the current study shed light on the promising anticancer activity of Ginkgo biloba leaves extract in treatment of hepatocellular carcinoma induced chemically in the experimental model through its apoptotic and antiproliferative properties.

Expanding the mutation and clinical spectrum of Roberts syndrome.

Roberts syndrome and SC phocomelia syndrome are rare autosomal recessive genetic disorders representing the extremes of the spectrum of severity of the same condition, caused by mutations in ESCO2 gene. We report three new patients with Roberts syndrome from three unrelated consanguineous Egyptian families. All patients presented with growth retardation, mesomelic shortening of the limbs more in the upper than in the lower limbs and microcephaly. Patients were subjected to clinical, cytogenetic and radiologic examinations. Cytogenetic analysis showed the characteristic premature separation of centromeres and puffing of heterochromatic regions. Further, sequencing of the ESCO2 gene identified a novel mutation c.244_245dupCT (p.T83Pfs*20) in one family besides two previously reported mutations c.760_761insA (p.T254Nfs*27) and c.764_765delTT (p.F255Cfs*25). All mutations were in homozygous state, in exon 3. The severity of the mesomelic shortening of the limbs and craniofacial anomalies showed variability among patients. Interestingly, patient 1 had abnormal skin hypopigmentation. Serial fetal ultrasound examinations and measurements of long bones diagnosed two affected fetuses in two of the studied families. A literature review and case comparison was performed. In conclusion, we report a novel ESCO2 mutation and expand the clinical spectrum of Roberts syndrome.

Implications of Sex Hormone Receptor Gene Expression in the Predominance of Hepatocellular Carcinoma in Males: Role of Natural Products.

The present study was planned to investigate the role of sex hormone receptor gene expression in the pathogenesis of hepatocellular carcinoma (HCC). Adult male Wistar rats were divided into seven groups. Group (1) was negative control. Groups (2), (5), (6), and (7) were orally administered with N-nitrosodiethylamine for the induction of HCC, then group (2) was left untreated, group (5) was orally treated with curcumin, group (6) was orally treated with carvacrol, and group (7) was intraperitoneally injected with doxorubicin, whereas groups (3) and (4) were orally administered only curcumin and carvacrol, respectively. The HCC group showed significant upregulation in the androgen receptor (AR) and the estrogen receptor-alpha (ERα) gene expression levels in the liver tissue. On the contrary, HCC groups treated with either curcumin or carvacrol showed significant downregulation in AR and ERα gene expression levels in the liver tissue. In conclusion, the obtained data highlight that both AR and ERα but not estrogen receptor-beta (ERß) gene expression may contribute to the male prevalence of HCC induced in male rats. Interestingly, both curcumin and carvacrol were found to have a promising potency in alleviating the male predominating HCC.

Curcumin: a unique antioxidant offers a multimechanistic approach for management of hepatocellular carcinoma in rat model.

This study was designed to investigate the role of curcumin against hepatocellular carcinoma (HCC) induced in rats. Forty rats were divided into five groups. Group (1) was negative control. Groups (2), (4), and (5) were orally administrated N-nitrosodiethylamine for HCC induction, then group (2) was left untreated, and group (4) was treated orally with curcumin, while group (5) was intraperitoneally injected with doxorubicin. Group (3) was served as curcumin control group. Serum alpha-fetoprotein, alpha L-fucosidase and vascular endothelial growth factor levels were analyzed. Gamma glutamyl transferase (GGT) and heat shock protein gp96 (HSPgp96) gene expressions were detected by RT-PCR. The immunohistochemical analysis of proliferating cell nuclear antigen (PCNA) and Ki-67 expressions was performed. Apoptosis was detected using DNA fragmentation assay. Also, histological investigation of liver tissue was achieved. Untreated HCC group showed significant elevation in the studied biochemical markers and significant upregulation in GGT and HSPgp96 gene expression as well as marked increase in PCNA and Ki-67 expression. Furthermore, this group revealed no DNA fragmentation. Histological investigation of liver tissue sections in HCC group revealed a typical anaplasia. On the other hand, the curcumin-treated group showed a significant depletion in the studied tumor markers and a significant downregulation in GGT and HSPgp96 gene expression. Also, this group displayed remarkable decrease in PCNA and Ki-67 expression. Moreover, this group revealed an obvious DNA fragmentation. Interestingly, treatment with curcumin showed remarkable improvement in the histological features of liver tissue. This study revealed the promising therapeutic role of curcumin against hepatocellular carcinoma owing to its antiangiogenic, antiproliferative, and apoptotic effects.

Potent neuroprotective role of novel melatonin derivatives for management of central neuropathy induced by acrylamide in rats.

Acrylamide (ACR) has been shown to be a neurotoxic agent for both laboratory animals and human. The present study aimed at synthesizing new functionalized melatonin derivatives bearing promising heterocyclic moiety that could be expected to have protective effect against ACR-induced neurotoxicity in adult female rats. The novel melatonin derivatives 4, 6, 7 and 11 were synthesized and their chemical structures were confirmed by studying their analytical and spectral data. The administration of ACR [i.p., 50 mg kg(-1) body weight (b. wt.)] alone resulted in significant increase in brain malondialdehyde level (MDA) and lactate dehydrogenase (LDH) activity whereas it caused significant decrease in brain monoamines levels and antioxidant enzymes activity. Treatment with melatonin derivatives 4, 6, 7 and 11 (i.p., 50 mg kg(-1) b. wt) prior to ACR produced significant decrease in brain MDA level and LDH activity with concomitant significant increase in brain monoamines and antioxidant enzymes activity. It could be concluded that the new synthesized melatonin derivatives exhibited promising protective activity against ACR-induced neurotoxicity.

Serodiagnosis of human intestinal schistosomiasis.

We developed an enzyme linked-immunosorbent assay (ELISA) for serodiagnosis of Schistosoma mansoni infection using a purified immunogenic fraction from schistosome adult worm, obtained by SDS-polyacrylamid gel electrophoresis. Sera from patients with active schistosomiasis (egg passers; n=10); inactive schistosomiasis previously treated with praziquantel (not passing eggs; n=10); fascioliasis, hydatosis (n=5); and healthy controls (n=10) were examined. Western blot analysis revealed that the Sm 31/32 KDa fraction of Schistosoma mansoni is recognized by sera from of both active and inactive schistosomiasis. ELISA IgG reactivity (optical density, OD) to Sm 31/32 KDa fraction by ELISA was significantly higher in sera of schistosomiasis patients (active and inactive), (p<0.001) compared to normal controls, while no significant difference was detected between active (OD=0.79 +/- 0.23) & inactive (OD=0.87 +/- 0.37) patients. No reactivity was detected using facioliasis or hydatosis sera. The overall level of specificity and sensitivity attained was 90% and 93%, respectively. It is concluded that the developed Sm 31/32 KDa ELISA may be of value in serodiagnosis of active and inactive intestinal Schistosoma mansoni infection in humans.