Plasma metadherin mRNA expression in bladder cancer.

Urinary bladder cancer (BC) is the ninth most common cancer worldwide. At present, the clinical diagnosis of BC depends on self-reported symptoms, tissue biopsy specimens by cystoscopy and from voided urine cytology. However, cystoscopy is an invasive examination and voided urine cytology has low sensitivity, which might provoke misdiagnosis. The search for cancer biomarkers in blood is worthy of intense attention due to patients' comfort and ease of sampling. This work aimed to study expression of mRNA metadherin (MTDH) in plasma, serum BC specific antigen 1 (BLCA-1) and cystatin C as biomarkers of BC and their relation to different disease stages. This study included 59 BC patients, 11 patients with benign bladder lesion and 18 subjects as normal controls. MTDH expression was assessed by real time polymerase chain reaction, BLCA-1, and cystatin C by the enzyme linked immunosorbent assay. The three biomarkers were elevated in BC patients than patients with benign bladder diseases and controls. Patients with BC grade 3 and 4 had higher cystatin C, BLCA-1 and MTDH in comparison to patients with grade 1 and grade 2 (p=0.000). The receiver operating characteristic curve analysis showed that BLCA-1 at a cutoff point of 32.5 ng/ml and area under the curve of 1.00, had 100% accuracy, 100% sensitivity, 100% specificity, 100% positive predictive values and 100% negative predictive value. In conclusion, BLCA-1 was a better biomarker than cystatin C and MTDH. Cystatin C, BLCA-1 and MTDH levels, can differentiate between benign bladder lesion and BC and correlated with tumor grades.especially with OL-HDF compared to HF-HD, with acceptable albumin loss in the dialysate.

Long noncoding RNA HOTAIR and Midkine as biomarkers in thyroid cancer.

Thyroid cancer is the most common endocrine malignancy, and its incidence is increasing. Differentiated thyroid cancer is the most common type and papillary thyroid carcinoma is the most common type of differentiated thyroid cancer. This work aimed to study long noncoding (Lnc) RNA homeobox transcript antisense RNA (HOTAIR) expression in plasma and serum midkine, a heparin binding growth factor, as biomarkers of thyroid cancer. This study included 27 thyroid cancer patients, 29 patients with benign thyroid disease and 26 individuals as normal controls. HOTAIR expression was assessed by real time polymerase chain reaction and midkine by ELISA. These biomarkers were elevated in thyroid cancer patients than patients with benign thyroid diseases and controls. Patients with thyroid cancer stage III had higher midkine levels in comparison to those with stage-I and stage-II (p < 0.001). Patients with grade II had higher midkine in comparison to those with grade I (p < 0.001). Statistically significant elevation of HOTAIR expression was found in stage III and stage II (p=0.001), compared to stage I. However, no difference was observed between stage II and stage III (p=0.533). There was no difference in both biomarkers in different histopathological types of thyroid cancer. ROC analysis was used for detection of thyroid cancer, midkine had AUC of 0.95 at a cutoff 897.5 pg/ml with a sensitivity of 98.0%, and specificity of 81.5% (p < 0.001). HOTAIR had AUC of 1 at a cutoff 11.8-fold change with a sensitivity and specificity of 100 %, (p < 0.001). We concluded that HOTAIR has high sensitivity and specificity in detection of thyroid cancer. It was correlated with tumor stage but not with histopathological types.

Neutralizing antibody responses in asymptomatic close contacts of COVID-19 patients and in asymptomatic healthcare workers.

The identification of novel antibodies that could neutralize SARS-CoV-2 is one of the novel approaches to use in combating COVID-19. This study aimed to explore the level of neutralizing antibodies (NAbs) in asymptomatic close contacts of COVID-19 patients and asymptomatic healthcare workers. In vitro qualitative detection of serum antibodies of participants from both populations was done using an anti-SARS-CoV-2 immunoassay. The study included 107 participants, of which 59.8% were healthcare workers and 40.2% were family contacts of confirmed COVID-19 cases. Their median age was 22 years. The percentage of positivity and median titer for NAbs were significantly higher among family contacts than mong healthcare workers (P = 0.013 and < 0.001, respectively). We also measured C-reactive protein (CRP) levels and the median value of CRP was significantly higher in the family members who had been in contact with COVID-19 patients than in healthcare workers (P < 0.001). In the family contact group, there was a significant negative correlation between the absolute lymphocyte count and CRP (r = -0.409, P = 0.034). There was no significant correlation between neutralizing antibody titers and either CRP or absolute lymphocyte count (P > 0.05 for both). In conclusion, the indication of elevated NAb titers in asymptomatic family contacts could help lay the groundwork for further studies to explore the potential utility of these antibodies to provide future immunity from infection within a family as well as for potential use in general during passive antibody therapies for COVID-19 patients.

Genetic detection of AmpC beta-lactamase among gram negative isolates "A Single Center Experience''.

The misuse of ß-Lactam antibiotics results in major problem, microbial resistance against these antibiotics by expression of ß-lactamases, facing its use. AmpCs are one of the ß-lactamases which confer resistance to penicillins, cephalosporins, cephamycins, and aztreonam, and are not affected by classic ß-lactamase inhibitors. Plasmid-mediated AmpC ß-lactamases pose a major challenge to infection control because the AmpC gene can be expressed in larger quantities and has a high transmissibility to other bacterial species. This study aimed to detect plasmid mediated AmpC ß-lactamases in gram negative isolates in Assiut university hospital. It was performed on 120 cefoxtin resistant isolates obtained from 300-gram negative isolates using the disc diffusion method as a screening test for AmpC production. Since the presence of pAmpC is often associated with the presence of ESBLs, phenotypic detection of ESBL was done using combined disc method and vitek2 compact 15. Phenotypic detection of AmpC was done by disc approximation method and inhibitor-based method using phenyl boronic acid (PBA). Genotypic detection of 5 plasmid mediated AmpC genes families (MOX, CIT, DHA, EBC, and FOX) was done by multiplex PCR. Our result showed that Klebsiella pneumoniae (62.5%) and Escherichia coli (25.8%) were the most frequent isolates. Only 15.8 %, 12.5%, 17.5% resistant isolates to cefoxitin were positive by using disc approximation test, inhibitor-based method using PBA (150 g/mL), and PBA (600 g/mL), respectively. Out of the 120 Cefoxitin-resistant isolates, 22 isolates (18.3 %) were positive by multiplex PCR. CIT and MOX were solely detected in 45.5% and 4.5%, respectively. CIT and FOX together were detected in 45.5%, CIT and DHA together in 4.5%. No isolate was positive for EBC gene. Finally, boronic acid test using 600 µg/mL PBA with 30µg ceftazidime, as phenotypic method for detecting AmpC ß- lactamases, was ranked very good for marking negative tests.

Correlation Between Vascular Endothelial Growth Factor and Long-Term Occurrence of HCV-Related Hepatocellular Carcinoma After Treatment with Direct-Acting Antivirals.

We aimed to evaluate the correlation between vascular endothelial growth factor (VEGF) and long-term occurrence of hepatocellular carcinoma after HCV treatment with direct-acting antivirals (DAAs) and the HCC stage. Two groups with HCV-related liver cirrhosis and HCC were included: group 1, HCC following DAAs; group 2, HCC did not receive DAAs. The serum level of VEGF and HCC staging was evaluated. The duration between DAAs and HCC was 21.81 ± 11.66 months. Portal vein thrombosis (PVT) was observed more in group 1 (31%). VEGF was relatively elevated in group 1 compared to group 2. HCC patients after DAAs, showed elevated VEGF with frequent PVT.

Galactin-3 and brain natriuretic peptide versus conventional echocardiography in the early detection of cirrhotic cardiomyopathy.

BACKGROUND/AIMS: Cirrhotic cardiomyopathy (CCM) is defined as an abnormal heart structure and function in cirrhotic patients. CCM includes systolic and diastolic dysfunction, electrophysiological abnormalities, and structural changes, both microscopic and macroscopic. Currently, there is no one diagnostic test that can identify patients with CCM. Evaluation of the validity of galactin-3 and brain natriuretic peptide (BNP) as biomarkers in the early detection of CCM in comparison to conventional echocardiography. MATERIALS AND METHODS: A case control study was carried out in the Departments of internal medicine and tropical Medicine, Assuit University, Egypt. Seventy-one subjects were divided into the following three groups: 26 cirrhotic patients without ascites, 25 cirrhotic patients with ascites, and 20 healthy controls. All groups underwent clinical examination, and laboratory investigation including BNP, galactin-3, and echocardiography. RESULTS: There was a significant difference between the three groups (p < 0.001) with regard to corrected QT (cQT), BNP and galactin-3. Left ventricular diastolic dysfunction with different grades was the most recorded cardiac abnormality in the patient group I and II (88.5% and 96%; respectively) with significantly increased frequency and severity in ascetic patients and with the advancement of liver cirrhosis. BNP and galactin-3 were sensitive and specific biomarkers for the detection of diastolic dysfunction in cirrhotic patients (77.6%, 95.5%, 89.9% and 86.4%; respectively). CONCLUSION: Diastolic dysfunction is a common cardiac abnormality in cirrhotic patients that worsens with the advancement of cirrhosis. BNP and galactin-3 had higher sensitivity and specificity in the early detection of CCM compared with those of conventional echocardiography.

Vitamin D status and viral response to therapy in hepatitis C infected children.

AIM: To study the frequency of vitamin D deficiency in patients with hepatitis C virus (HCV) infection and to evaluate the role of vitamin D supplementation in improving antiviral therapy. METHODS: Sixty-six children aged from 7-14 years (mean ± SD, 11.17±2.293) diagnosed with HCV infection were matched to 28 healthy controls. Serum levels of 25 (OH) D3, calcium, phosphorus, alkaline phosphatase and plasma level of parathormone were measured. Quantitative PCR for HCV was performed Bone density was determined by dual energy X-ray absorptiometry. All cases received conventional therapy, and only 33 patients received vitamin D supplementation. RESULTS: Children with HCV showed significantly increased levels of HCV RNA (P<0.001), parathormone (P<0.01) and decreased vitamin D levels (P<0.05) (33.3% deficient and 43.3% insufficient) compared with controls. Abnormal bone status (Z score -1.98±0.75) was found in ribs, L-spine, pelvis and total body. Cases treated with vitamin D showed significant higher early (P<0.04) and sustained (P<0.05) virological response. There was a high frequency of vitamin D deficiency among the Egyptian HCV children, with significant decrease in bone density. The vitamin D level should be assessed before the start of antiviral treatment with the correction of any detected deficiency. CONCLUSION: Adding vitamin D to conventional Peg/RBV therapy significantly improved the virological response and helped to prevent the risk of emerging bone fragility.

Vitamin D level and Fok-I vitamin D receptor gene polymorphism in Egyptian patients with type-1 diabetes.

Many cellular, preclinical and observational studies support a role of vitamin D in pathogenesis of type-1 diabetes mellitus (DM). The vitamin D receptor (VDR) locus has been studied in different populations for association with susceptibility to immune-mediated diseases, but with inconsistent findings in type-1 DM. This study aimed to investigate vitamin D status in patients with type-1 DM. We examined the frequency of VDR Fokl (rs10735810) gene polymorphism, and its association with serum 25-hydroxyvitamin D (25(OH) D) level in Egyptian patients with type-1 DM. 132 children with type-1 DM and 40 age and sex matched healthy control subjects were studied. VDR Fokl polymorphism was assessed using polymerase chain reaction and restriction fragment length polymorphism (RFLP) analysis. Diabetic children demonstrated lower circulating levels of 25(OH) D than the controls (13.4 +/- 7.6 vs 32.1 +/- 3.8ng/ml) (P < 0.01). Patients with deficient 25(OH) D showed lower calcium levels and higher HbA1c% than those with sufficient levels (8.1 +/- 2.1 versus 9.1 +/- 1.6 mg/dl & 9.9 +/- 2.5 versus 8.1 +/- 1.4%, respectively (P < 0.05). There was no significant difference in the genotype distribution or the allele frequencies of VDR Fokl between patients and controls. The odds ratio (OR) was 1.08 (P = 0.76), and the 95% confidence interval (CI) ranged from 0.64-1.85. The diabetic carriers of the ff genotype showed low serum levels of 25(OH) D and calcium when compared with the carriers of the F allele (9.1 +/- 4.4 vs 13.1 +/- 7 and 13.9 +/- 6.09 ng/ml & 8.1 +/- 2.1 vs 9.1 +/- 1.1 and 9.3 +/- 1.2 mg/dl, respectively) (P < 0.05). In conclusion, 84.8% of children with type-1 DM have low circulating levels of 25(OH) D. These patients have poor glycemic control (56.06%) than those with sufficient levels of 25(OH) D. Fokl polymorphism of VDR gene is associated with vitamin D deficiency but has no significant role in susceptibility to type-1 diabetes.