Multiplex PCR: Aid to more-timely and directed therapeutic intervention for patients with infectious gastroenteritis.

BACKGROUND: Multiplex PCR is a sensitive and rapid method compared with conventional methods. Therefore, we use multiplex PCR for the rapid detection of the four major intestinal pathogens causing gastroenteritis (Shigella spp., Campylobacter spp., Aeromonas spp. and Enterohemorrhagic Escherichia coli [EHEC]) in stool specimens. MATERIALS AND METHODS: A prospective randomized study using 200 stool samples obtained from patients presented with acute gastroenteritis during the study period (between February 2019 and December 2021). Bacteria in stool samples were identified using conventional culture methods and multiplex PCR for stool samples. RESULTS: The identified organisms using conventional cultures; were Shigella (27%), Aeromonas species (10%) and EHEC (O157) (8%). Using multiplex PCR. Shigella spp. was the most commonly identified pathogen (detected in 40.5% of positive samples), followed by Aeromonas spp. (30%), EHEC (20%) and Campylobacter species was only detected in (1%) of positive samples. The diagnostic evaluation of multiplex PCR in relation to conventional method in diagnosis of Shigella, EHEC and Aeromonas showed, sensitivity of 100% (for each), specificity of 88.5%, 92.4%, 77.8% respectively. However, the diagnostic evaluation of multiplex PCR in relation to conventional method in diagnosis of Campylobacter showed specificity of 99% and NPV of 100%. CONCLUSIONS: Multiplex PCR is an accurate and rapid method for detection of common intestinal pathogens causing severe gastroenteritis. a rapid method that could be used in outbreaks for diagnosis of the common enteric pathogens causing fatal gastroenteritis.

Usefulness of Circulating Methylated p16 as a Noninvasive Molecular Biomarker for Hepatitis C-Related Hepatocellular Carcinoma with Normal Serum Alpha-Fetoprotein Levels.

BACKGROUND: Screening of hepatocellular carcinoma (HCC) is challenged especially in patients with normal alpha-fetoprotein (AFP) levels. Aberrant p16 methylation has been implicated in HCC. OBJECTIVES AND AIMS: This study aimed to assess serum methylated p16 (MP16) expression levels and to evaluate MP16 diagnostic performance in HCC detection among HCV-infected Egyptian patients with normal AFP levels. METHODS: MP16 levels were quantified using real-time PCR in 230 serum samples (30 healthy controls, 95 with HCV-HCC, 40 with chronic hepatitis C "CHC" and 65 with HCV cirrhosis). Diagnostic performance of MP16 for diagnosis of HCC was done using receiver operator characteristic curve analysis. RESULTS: Serum MP16 levels were significantly higher in HCC than CHC, cirrhosis, and healthy subjects and significantly higher in HCC with normal AFP levels than those with higher AFP. ROC curves revealed promising diagnostic performance for MP16 in discriminating HCC with normal AFP levels from non-HCC cases. This predictive ability improved by combining MP16 and AFP (AUC of 0.872 with 100% sensitivity, 76.5% specificity, 79.1% positive predictive value, 100% negative predictive value, and 87.5% accuracy). CONCLUSION: MP16 can be a potential noninvasive molecular biomarker for HCC detection in patients with hepatic mass(es) and normal AFP levels especially in those where liver biopsy and radiological imaging cannot be done.

Validity of serum amyloid A and HMGB1 as biomarkers for early diagnosis of gastric cancer.

BACKGROUND AND AIM: Gastric carcinomais a frequent neoplasm with poor outcome, and its early detection would improve prognosis. This study was designed to evaluate the possible use of new biomarkers, namely SAA and HMGB1, for early diagnosis of gastric cancer. METHODS: A total of 100 patients presenting with gastric symptoms were included. All patients underwent upper endoscopic evaluation, histopathological diagnosis and serum CEA, SAA, and HMGB1 measurements. RESULTS: Patients were classed endoscopically with neoplastic, inflammatory, and normal-appearing gastric mucosa: 50, 25, and 25 patients, respectively. Histologically, half the patients had chronic gastritis and the remaining cases gastric carcinoma of diffuse (n=28) or intestinal (n=22) type. SAA at cutoff of 18.5 mg/L had the best validity to differentiate gastritis from gastric carcinoma, with AUC, sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) of 0.99, 98%, 100%, 100%, and 98%, respectively, followed by HMGB1 at cutoff of 14.5 pg/µL, with AUC, sensitivity, specificity, PPV, and NPV of 0.91, 70%, 96%, 94.6%, and 76.2%, respectively. Sensitivity, specificity, PPV, and NPV of serum CEA at cutoff of 2.9 ng/mL to differentiate gastritis from gastric carcinoma were 42%, 72%, 60%, and 55.4%, respectively, with AUC of 0.53. Nonetheless, higher serum levels of both SAA and HMGB1 reflected higher tumor grade (P=0.027 and P=0.016, respectively) and advanced tumor stage (P-OBrk-0.001 for both). CONCLUSION: Serum levels of both SAA and HMGB1 could be of great value for early diagnosis of gastric carcinoma, comparable to the diagnostic role of serum CEA, which is not valid for early diagnosis of gastric cancer.

Epidemiological Characteristics and Etiology of Budd-Chiari Syndrome in Upper Egypt.

BACKGROUND AND OBJECTIVES: Budd-Chiari syndrome (BCS) is a rare disorder caused by obstruction to hepatic venous outflow. It affects all races, usually during the third or fourth decade of life. Higher prevalence had being evident in developing countries. The aim of the present study was to clarify sociodemographic features, clinical, radiological presentations, and etiology of BCS among Upper Egyptian patients. PATIENTS AND METHODS: This retrospective cohort study enrolled 50 Upper Egyptian Patients with confirmed primary BCS. Liver, coagulation, and thrombophilia workup profiles were performed as anticardiolipin antibodies, lupus anticoagulant, protein C, protein S, and antithrombin III assays. Factor V Leiden and JAK2 mutations were assessed. Full radiological assessment was done. RESULTS: Fifty patients were included. There were 28 males (56%) and 22 females (44%) with mean age (32.5 ± 11.1 years). The etiological factor was not identified in 22% of cases (n=11). Isolated factor C deficiency was found in 26% (n=13) with male predominance 39.3% and protein S deficiency in 10% (n=5). Factor V Leiden mutation was the etiology in 5 patients (10%). Membranous web and antiphospholipid syndrome each were the etiology in 8% (n=4). Behςet's disease was diagnosed in 4% (n=2). Cases of liver cirrhosis(LC) were 41/50(82%)they were :33/50(66%) LC child class C, 8 /50(16%)  LC child class B, and 0/50 (0%) LC child class A. Abdominal pain was the most common symptom (96%), and ascites was the most common sign (82%). Obstruction of hepatic veins was present in 80%. CONCLUSION: BCS in Upper Egyptian patients was mainly occurred in males in the third and fourth decade of life, mostly with liver cirrhosis. The most common etiology is isolated protein C deficiency followed by Factor V Leiden mutation and isolated protein S deficiency. Hepatic veins obstruction was the most common pattern of vascular involvement.

Management Of Patients With Hepatitis B Virus Reactivation Post-DAA Treatment Of Chronic Hepatitis C Virus Infection In HCV-HBV Coinfected Patients With Pretreatment HBeAg Seroconversion And Early Degree Of Hepatic Fibrosis.

BACKGROUND AND AIM: Hepatitis C virus (HCV)-HBV coinfection is a significant health problem with rapid progression of liver disease without precise diagnosis and treatment. We aimed in this study to identify if there were any role of HBV antiviral therapy in patients with HBV reactivation after direct-acting antiviral therapy in HCV-HBV coinfected patients. METHODS: A prospective random study was carried out on 140 patients presenting with chronic HCV and chronic HBV coinfection. All patients had pretreatment HBeAg seroconversion, HBV DNA <2,000 IU/mL, normal liver enzymes, and F0/F1 hepatic fibrosis. They treated with sofosbuvir 400 mg and daklatasvir 60 mg once daily for 3 months. All patients underwent pretreatment hepatic fibrosis assessment using Fibro Scan and laboratory investigations: platelet count, liver-function tests, quantitative HCV PCR, HBsAg, HBc IgG, HBeAg, and HBeAb. All patients were followed up at 1, 3, 6, and 12 months from the start of HCV therapy. RESULTS: The study enrolled 140 HCV-HBV coinfected patients: 55% were F0 and the rest F1. All our patients had negative HCV PCR at 1 month posttreatment and had achieved sustained virologic response with negative HCV PCR 3 months after treatment end. Four patients showed HBV reactivation with raised HBV DNA PCR and liver enzymes. Their mean age was 23.7±2.7 years, and three were male. Regarding patients with HBV reactivation, at 12 months posttreatment they showed significant decreases in liver enzymes, bilirubin, and INR, with increased platelet count (P=0.001), each with undetectable HBV PCR (P=0.001). CONCLUSION: HBV-HCV coinfected patients with no/mild hepatic fibrosis, HBeAg seroconversion, and HBV DNA <2,000 IU/mL can complete direct-acting antiviral therapy without HBV antiviral treatment with close monitoring.

Types and microbiological spectrum of infections in patients with cirrhosis: A single-centre experience in Upper Egypt.

BACKGROUND AND STUDY AIMS: Egypt has a high prevalence of hepatitis C virus (HCV) and high morbidity and mortality related to cirrhosis complications. Patients with cirrhosis have an increased risk of bacterial infections. Approximately 25-35% of cirrhotics had infections at admission or during hospitalisation. Data on infection among cirrhotics in Egypt are limited. This study aimed to determine the frequency and microbiological spectrum of infections in cirrhotics and possible risk factors. PATIENTS AND METHODS: This study was conducted at a tertiary care hospital. The frequency and microbiological spectrum of infections in cirrhotics were determined. The risk factors for infection were evaluated. RESULTS: Of the 100 patients with liver cirrhosis, 61% had infection. Ascitic fluid infection (AFI) was the most common infection (44.3%), followed by urinary tract infection (UTI) (21.3%), respiratory tract infection (RTI) (19.7%), gastroenteritis (6.6%) and skin infection (4.9%). The only risk factor for infection among cirrhotics was diabetes mellitus (DM) (p=0.047). The mean value of mid-arm muscle circumference was significantly lower in the infected group (p=0.047). Among all the cirrhotics, 32.0% had mild to moderate malnutrition and 52.0% had severe malnutrition. The frequency of infection was higher in severe malnutrition (71.2%). CONCLUSIONS: The frequency of infections among cirrhotics was 61%. Many types of infections including AFI, RTI, UTI and skin infections were present in patients with liver cirrhosis, but AFI was the most common. DM was the only risk factor for infection, and independent predictors for infection were elevated WBC count and C-reactive protein levels. The frequency of infection was related to the degree of malnutrition.

VAP score as a novel non-invasive liver fibrosis model in patients with chronic hepatitis C.

AIM: Assessment of liver fibrosis in chronic hepatitis C (CHC) patients is necessary before antiviral treatment. This study aimed to evaluate the effectiveness of eight non-invasive models (aspartate aminotransferase [AST]/alanine transaminase ratio [AAR], AST/platelet ratio index [APRI], fibrosis-cirrhosis index [FCI], fibrosis index [FI], fibrosis-4 [FIB-4] score, fibrosis quotient [FibroQ], King, and von Willebrand factor antigen (vWF-Ag)/thrombocyte ratio [VITRO] scores) for predicting fibrosis compared with liver biopsy and to create a new score for predicting different fibrosis stages with increased accuracy. METHODS: We prospectively studied 127 treatment-naive CHC patients who underwent liver biopsy. The AAR, APRI, FCI, FI, FIB-4, FibroQ, King and VITRO scores were calculated and correlated with fibrosis stages. A new score (VAP) was derived from vWF-Ag, AST, and platelets: [VAP = (AST (U/L) × vWF-Ag)/platelets (109 /L)]. RESULTS: Apart from AAR, readily available scores were correlated with liver fibrosis stages. VITRO (r = 0.62) and APRI (r = 0.46) showed the closest correlation. Our new (VAP) score significantly correlated with fibrosis stages (r = 0.702, P < 0.001). Compared to other scores, VAP had the highest area under the receiver operating characteristic curve, with 0.854, 0.921, 0.849, and 0.861 for mild (F1), significant (≥F2), advanced (≥F3) fibrosis, and cirrhosis (F4) respectively. At a cut-off value >1, VAP had 75.2% sensitivity and 100% positive predictive value for predicting mild fibrosis. At a cut-off value >2.3 for predicting cirrhosis, VAP had 73% sensitivity and 81.7% positive predictive value. CONCLUSIONS: The VAP score is a novel model that had higher diagnostic performance to predict different fibrosis stages and subclinical cirrhosis among CHC patients compared to the other studied scores and hence may offer a useful strategy to stratify patients who would benefit from direct-acting antivirals.

SIRT-1expression is associated with expression of NANOG in patients with colorectal adenocarcinoma.

AIMS: The study aimed to investigate the quantitative expression of NANOG, p38 α , NCF2, ELF and TGF-ß genes in patients with colorectal adenocarcinoma, adenoma and normal colonic tissue and their correlation with SIRT-1 protein level expression. METHOD: This study enrolled one hundred sixty seven patients; group A: 87 patients with colonoscopic findings of no adenoma or adenocarcinoma and group B: 80 patients with colorectal mass. Consecutive colonoscopic examinations were conducted, and tissue samples were taken from the colonic lesions/masses. Total RNA was isolated and mRNA expression level of NANOG, mitogen activated p38α , Neutrophil Cytosol Factor 2 (NCF2), Embryonic Liver Fodrin (ELF) and Transforming Growth Factor Beta (TGF-ß) genes were quantified by qRT-PCR. Sirt-1 protein expression level was assessed by quantitative western blot. RESULTS: There were significantly high level of mRNA transcripts expression of the genes studied in patients with adenocarcinoma and adenoma compared with normal tissue (P value < 0.01), NANOG, NCF2, ELF and TGF-ß at a cut of > 0.314, > 0.392, 0.349 and 0.333 respectively showed sensitivity (96.5%, 98.8%, 95.3%, 98.8%) and specificity of (95.3%, 92.6%, 89.5%, 93.8%) respectively in diagnosing colonic adenocarcinoma. Sirt-1 protein level was significantly highly expressed in colorectal adenocarcinoma compared to normal and adenoma colonic tissue and positively correlated with NANOG. CONCLUSION: Over expression of NANOG, p38α , NCF2, ELF and TGF-ß genes in both cases of adenocarcinoma and adenoma could have a diagnostic value. SIRT-1 and NANOG are high correlated biological markers for diagnosis and prognosis follow up in patients with adenocarcinoma.

Multidisciplinary decision making in the management of hepatocellular carcinoma: A hospital-based study.

BACKGROUND/AIMS: To evaluate the short-term outcome of the decision taken by the Hepatoma Board for the treatment of Hepatocellular carcinoma (HCC). MATERIALS AND METHODS: This was a prospective descriptive study involving 74 patients with HCC diagnosed by the known criteria. The decisions taken by the Hepatoma Board for the 74 patients were as follows: 1- surgical resection (7 patients), 2- local ablative therapy (LAT) (22 patients), 3- conventional transarterial chemoembolization (TACE) (24 patients), and 4- palliative supportive care (21 patients). RESULTS: The short-term mortality rate was 25.7% of the total patients. The success rate was nearly equal in LAT (68.2%) and surgery (71.4%), whereas the success rate was approximately 33.3% in TACE. There was no difference in the mean total bilirubin level before and after LAT, surgery, or TACE (p>0.05 for each). There was a significant decrease in the mean serum albumin level after TACE (p=0.000). There was a decrease in the mean alpha fetoprotein level after surgery and LAT (p=0.033) for surgery and (p=0.048) for LAT. CONCLUSION: The management of HCC is better performed through a multidisciplinary team decision. Surgery has comparable outcome to LAT but is more invasive. According to our local experience, conventional TACE has a success rate of 33.3%.