Role of Lymphotoxin-α Gene Polymorphism in Hepatitis C Virus-Related Chronic Liver Disorders.

BACKGROUND: Tumor necrosis factor (TNF) family includes lymphotoxin-alpha (LTA) which is a pro-inflammatory cytokine which plays a role in hepatic fibrogenesis. LTA gene polymorphism plays a role in different inflammatory and immunomodulatory diseases. This polymorphism is also suggested to affect chronic hepatitis C (CHC) infection course. AIM: To study the contribution of LTA gene polymorphism in different chronic hepatitis C stages and hepatocellular carcinoma risk. PATIENTS AND METHODS: Our study included 108 chronic HCV patients grouped according to the disease stage. Group (A): CHC, group (B): liver cirrhosis (LC), group (C): LC with HCC, and group (D): healthy controls. Routine laboratory investigations, polymerase chain reaction (PCR) for quantification of HCV, abdominal ultrasonography, and Liver stiffness measurement (LSM) were done. Child-Turcotte-Pugh, Model for end-stage liver disease (MELD), and Fibrosis index based on 4 (FIB-4) scores were calculated. We used the PCR-restriction fragment length polymorphism technique for lymphotoxin-α genotyping. RESULTS: The A/G genotype was predominant in all groups. In HCC patients, G/G genotype was more frequent (31.8%) than in the LC group (19.4%), CHC group (17.8%), and controls (4.17%). A significant association was found between LTA genotypes and the child classes in HCC (P<0.01) but not in LC patients (P>0.05). HCC patients carrying A/G genotype had higher MELD scores than other genotypes. Multivariate binary logistic regression analysis confirmed that LTA G/G genotype and low platelet count were independent predictors for HCC development in patients with HCV-related LC. CONCLUSION: Detection of LTA G/G genotype in chronic HCV patients could help to recognize high-risk patients for disease progression and HCC development.

Identification of Fungal Pathogens in Otomycosis and Their Drug Sensitivity: Our Experience

Abstract Introduction Otomycosis is a common problem in otolaryngology practice. However, we usually encounter some difficulties in its treatment because many patients show resistance to antifungal agents, and present high recurrence rate. Objectives To determine the fungal pathogens that cause otomycosis as well as their susceptibility to the commonly used antifungal agents. Additionally, to discover the main reasons for antifungal resistance. Methods We conducted an experimental descriptive study on 122 patients clinically diagnosed with otomycosis from April 2016 to April 2017. Aural discharge specimens were collected for direct microscopic examination and fungal culture. In vitro antifungal susceptibility testing was performed against the commonly used antifungal drugs. We tested the isolated fungi for their enzymatic activity. Results Positive fungal infection was found in 102 samples. The most common fungal pathogens were Aspergillus and Candida species, with Aspergillus niger being the predominant isolate (51%). The antifungal susceptibility testing showed that mold isolates had the highest sensitivity to voriconazole (93.48%), while the highest resistance was to fluconazole (100%). For yeast, the highest sensitivity was to nystatin (88.24%), followed by amphotericin B (82.35%), and the highest resistance was to terbinafine (100%), followed by Itraconazole (94.12%). Filamentous fungi expressed a high enzymatic ability, making them more virulent. Conclusion The Aspergillus and Candida species are the most common fungal isolates in otomycosis. Voriconazole and Nystatin are the medications of choice for the treatment of otomycosis in our community. The high virulence of fungal pathogens is owed to their high enzymatic activity. Empirical use of antifungals should be discouraged.

Identification of Fungal Pathogens in Otomycosis and Their Drug Sensitivity: Our Experience.

Introduction Otomycosis is a common problem in otolaryngology practice. However, we usually encounter some difficulties in its treatment because many patients show resistance to antifungal agents, and present high recurrence rate. Objectives To determine the fungal pathogens that cause otomycosis as well as their susceptibility to the commonly used antifungal agents. Additionally, to discover the main reasons for antifungal resistance. Methods We conducted an experimental descriptive study on 122 patients clinically diagnosed with otomycosis from April 2016 to April 2017. Aural discharge specimens were collected for direct microscopic examination and fungal culture. In vitro antifungal susceptibility testing was performed against the commonly used antifungal drugs. We tested the isolated fungi for their enzymatic activity. Results Positive fungal infection was found in 102 samples. The most common fungal pathogens were Aspergillus and Candida species, with Aspergillus niger being the predominant isolate (51%). The antifungal susceptibility testing showed that mold isolates had the highest sensitivity to voriconazole (93.48%), while the highest resistance was to fluconazole (100%). For yeast, the highest sensitivity was to nystatin (88.24%), followed by amphotericin B (82.35%), and the highest resistance was to terbinafine (100%), followed by Itraconazole (94.12%). Filamentous fungi expressed a high enzymatic ability, making them more virulent. Conclusion The Aspergillus and Candida species are the most common fungal isolates in otomycosis. Voriconazole and Nystatin are the medications of choice for the treatment of otomycosis in our community. The high virulence of fungal pathogens is owed to their high enzymatic activity. Empirical use of antifungals should be discouraged.

Neutrophil CD11b, CD64 and Lipocalin-2: Early Diagnostic Markers of Neonatal Sepsis.

Neonatal sepsis remains a global health problem particularly in the developing countries. Its diagnosis remains one of the most difficult issues in clinical medicine. Many immunologic markers including neutrophils CD11b and CD 64 and Lipocalin-2 have been tested as biomarkers of neonatal sepsis. The aim of the present study was to assess the value of these markers for early diagnosis of neonatal sepsis. The study included 60 neonates with suspected neonatal sepsis and 20 apparently healthy controls. Lipocalin-2 serum level was assessed by ELISA while neutrophils CD11b and CD64 expressions were evaluated by flow cytometry. Neutrophils CD64 and CD11b expression levels elevated significantly in cases (67.8±7.57) and (57.01±2.46) respectively than in controls (11.78±7.20) and (8.26±4.79). Lipocalin-2 serum level was significantly higher in the patients (145.3 ± 55.3) than in controls (22.4 ± 12.9). In conclusion, neutrophils CD64, CD11b and Lipocalin-2 are early, specific and sensitive diagnostic markers of neonatal sepsis.

Pentraxin 3 Genetic Variants and The Risk of Active Pulmonary Tuberculosis.

Tuberculosis is a major health problem worldwide. Genetic factors are considered important determinants of the host susceptibility to Mycobacterium tuberculosis. The aim of the current study was to evaluate the association between pentraxin 3 genetic variants and the susceptibility and severity of active pulmonary tuberculosis. The study included 100 patients with newly diagnosed pulmonary TB and 50 healthy controls. PTX3 plasma level was assayed using ELISA and PTX3 genotypes (rs2305619, rs3816527, rs1840680) were detected by real time PCR in all participants. PTX3 rs1840680 genotype (AA) and allele (A) were significantly higher in the study group while the genotype (GG) was higher in the control group. The plasma level of PTX3 was higher in the patients than controls (P < 0.0001). There is a strong association between PTX3 plasma level and the activity and severity of pulmonary TB. PTX3 rs1840680 genotype AA is associated with increased risk of active pulmonary TB.

Interleukin-10 and Interferon Gamma Gene Polymorphisms and Hepatitis C Virus-Related Liver Cirrhosis Risk.

The aim of the study was to evaluate the association between the gene polymorphisms in interleukin-10 (IL-10) and interferon gamma (IFN-γ) genes with susceptibility and severity of hepatitis C virus (HCV) infection among Egyptian patients. Interleukin-10 -592 A/C, -1082 G/A and IFN-γ +874 T/A genotypes were determined in 100 chronic HCV patients and 50 healthy controls using restriction fragment length polymorphism (RFLP) and the amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) respectively. IL-10 -592 A/C polymorphism genotyping revealed that the frequency of CC genotype was significantly higher in chronic HCV patients than in controls (58% versus 30%, P < 0.05). Regarding IL-10 -1082 G/A polymorphism genotyping, a higher frequency of GG genotype was found in chronic HCV patients compared to controls (31% versus 10%, P < 0.05). IFN-γ +874 T/A genotyping showed that TT genotype was significantly higher in chronic HCV participants than controls (31% versus 18%, P < 0.05), while a higher frequency of T allele was found in cirrhotic patients compared to noncirrhotic patients (P < 0.05). Our observations suggested that IL-10 -592 A/C, -1082 G/A, and IFN-γ +874 T/A polymorphisms had a strong association with susceptibility to HCV infection. However, no significant association was observed between the cytokines (IL-10 and IFN-γ) genotypes profile and HCV-liver cirrhosis risk in the studied population, except for the high frequency of IFN-γ +874 T allele in cirrhotic patients.

The Role of Bcl-2 and Bax as Markers of Disease Progression in Hepatitis C Virus Infected Patients.

Many apoptotic markers have been linked to hepatic cell injury in HCV-related liver diseases, and hence could be used as potential markers for early detection of the disease. The present study aimed to assess the role of apoptotic markers Bcl-2 and Bax in the pathogenesis of chronic HCV-related liver diseases. A total of 85 participants were enrolled into the study; 70 chronic HCV patients (35 non-cirrhotic and 35 cirrhotic), and 15 healthy controls. The serum levels of Bcl-2 and Bax were assayed in all participants by ELISA. Bcl-2 and Bcl-2/Bax ratio were significantly higher in non-cirrhotic patients than the cirrhotic and controls (P < 0.001). Bax was significantly higher in cirrhotic patients compared to the other groups (P < 0.001). Positive and negative correlations were found between serum Bcl-2, Bax, Bcl-2/Bax ratio and HCV viral load in non-cirrhotic and cirrhotic patients respectively. These findings provide an evidence that apoptosis is dysregulated in patients with chronic HCV.

Serum MicroRNA-122 and MicroRNA-155: Markers of Disease Progression in Hepatitis C viral infection.

Chronic HCV with its longstanding complications of cirrhosis and HCC is a highly prevalent and challenging problem in Egypt. Recently, microRNAs are ranked as potential biomarkers for early diagnosis of HCV related complications. The aim of the present study was to evaluate the role of miRNA-122 and miRNA-155 for prediction of progression of HCV infection and for diagnosis of HCC. A total of 92 chronic HCV patients [chronic HCV (group 1, n =32); chronic HCV with cirrhosis (group 2, n=31); chronic HCV with HCC (group 3, n=29)] were enrolled into the study. Expression of serum miRNA-122 and miRNA-155 was assayed by real-time PCR in all participants. The serum level of miR-122 was significantly higher in chronic HCV patients than in healthy controls and both of cirrhotic and HCC patients (P < 0.001). Serum miR-155 was significantly elevated in HCC than in controls and non-HCC patients (P < 0.001). MiR-155 at the cut-off value of >6.11 for HCC diagnosis, had sensitivity and specificity of 72.4% and 95.2%, respectively. In conclusion; microRNA-122 is a potential marker of progression of hepatocytes injury in patients infected with HCV but not a reliable marker for diagnosis of HCC. MicroRNA-155 is a relatively reliable marker for HCC detection.

Flow cytometric analysis of peripheral blood lymphocytes in patients with vitiligo.

Vitiligo is an acquired cutaneous disorder characterized by progressive and selective destruction of melanocytes. Although, the exact etiology of vitiligo is still obscure, autoimmunity is strongly implicated in its pathogenesis. The aim of this study was to evaluate the immunological alterations in the peripheral lymphocytes in patients with vitiligo and their possible immunomodulation by narrowband ultraviolet B phototherapy. The study comprised of 44 patients with vitiligo (23 untreated, 21 treated) and 20 normal control subjects who were studied for peripheral blood lymphocytes imbalance using flow cytometry. The percentages of total T-lymphocytes, B-lymphocytes, helper T cells, cytotoxic T cells, activated T-lymphocytes, and natural killer cells were evaluated with the use of CD3, CD19, CD4, CD8, CD25, HLA-DR and CD56 monoclonal antibodies, respectively. A statistically significant lower difference in the median values of CD4+ cells (helper T lymphocytes) and CD4+/CD8+ ratio was found between untreated and both of treated and control groups. Natural killer cells (CD3-CD56+) were significantly higher in untreated than the other two groups. On the other hand, activated T-helper cells (CD4+CD25+) were significantly higher in both untreated and treated groups than the control subjects. In conclusion, the reported immunological alterations supported the role of cellular autoimmune mechanisms in the pathogenesis of the disease. Narrowband ultraviolet B phototherapy may enhance cellular immunity in vitiligo patients.