Dina M. Abdallah

Elucidation of the mechanism of atorvastatin-induced myopathy in a rat model.

Myopathy is among the well documented and the most disturbing adverse effects of statins. The underlying mechanism is still unknown. Mitochondrial dysfunction related to coenzyme Q10 decline is one of the proposed theories. The present study aimed to investigate the mechanism of atorvastatin-induced myopathy in rats. In addition, the mechanism of the coenzyme Q10 protection was investigated with special focus of mitochondrial alterations. Sprague-Dawely rats were treated orally either with atorvastatin (100mg/kg) or atorvastatin and coenzyme Q10 (100mg/kg). Myopathy was assessed by measuring serum creatine kinase (CK) and myoglobin levels together with examination of necrosis in type IIB fiber muscles. Mitochondrial dysfunction was evaluated by measuring muscle lactate/pyruvate ratio, ATP level, pAkt as well as mitochondrial ultrastructure examination. Atorvastatin treatment resulted in a rise in both CK (2X) and myoglobin (6X) level with graded degrees of muscle necrosis. Biochemical determinations showed prominent increase in lactate/pyruvate ratio and a decline in both ATP (>80%) and pAkt (>50%) levels. Ultrastructure examination showed mitochondrial swelling with disrupted organelle membrane. Co-treatment with coenzyme Q10 induced reduction in muscle necrosis as well as in CK and myoglobin levels. In addition, coenzyme Q10 improved all mitochondrial dysfunction parameters including mitochondrial swelling and disruption. These results presented a model for atorvastatin-induced myopathy in rats and proved that mitochondrial dysfunction is the main contributor in statin-myopathy pathophysiology.

Adalimumab ameliorates OVA-induced airway inflammation in mice: role of CD4(+) CD25(+) FOXP3(+) regulatory T-cells.

Asthma is a chronic inflammatory heterogeneous disorder initiated by a dysregulated immune response which drives disease development in susceptible individuals. Though T helper 2 (TH2) biased responses are usually linked to eosinophilic asthma, other Th cell subsets induce neutrophilic airway inflammation which provokes the most severe asthmatic phenotypes. A growing evidence highlights the role of T regulatory (Treg) cells in damping abnormal Th responses and thus inhibiting allergy and asthma. Therefore, strategies to induce or augment Treg cells hold promise for treatment and prevention of allergic airway inflammation. Recently, the link between Tumor necrosis factor-α (TNF-α) and Treg has been uncovered, and TNF-α antagonists are increasingly used in many autoimmune diseases. Yet, their benefits in allergic airway inflammation is not clarified. We investigated the effect of Adalimumab, a TNF-α antagonist, on Ovalbumin (OVA)-induced allergic airway inflammation in CD1 mice and explored its impact on Treg cells. Our results showed that Adalimumab treatment attenuated the OVA-induced increase in serum IgE, TH2 and TH1 derived inflammatory cytokines (IL-4 and IFN-γ, respectively) in bronchoalveolar lavage (BAL) fluid, suppressed recruitment of inflammatory cells in BAL fluid and lung, and inhibited BAL fluid neutrophilia. It also ameliorated goblet cell metaplasia and bronchial fibrosis. Splenocytes flow cytometry revealed increased percentage of CD4(+) CD25(+) FOXP3(+) Treg cells by Adalimumab that was associated with increase in their suppressive activity as shown by elevated BAL fluid IL-10. We conclude that the beneficial effects of Adalimumab in this CD1 neutrophilic model of allergic airway inflammation are attributed to augmentation of Treg cell number and activity.

Toll like receptor 3 expression as a novel predictor of response to treatment in chronic hepatitis C virus patients

Abstract Purpose. This work was carried out to study the level of pretreatment hepatic expression of Toll like receptor 3 (TLR3) among chronic HCV patients, aiming to determine if there are consistent differences in gene expression, between those who show complete early virological response (cEVR) at week-12 of Pegylated-Interferon α-2a plus ribavirin treatment and others who are not responding to this combination, also if this could be used to predict treatment outcomes. Method. A total of 61 chronic hepatitis C patients were enrolled in the study. For all of them, baseline hepatitis C virus (HCV) viral load was determined and TLR3 gene expression was examined in their hepatic percutaneous needle biopsy specimens using a real time–polymerase chain reaction technique. Hepatic TLR3 was also traced using immunohistochemistry. All patients followed a 12-week regimen of Pegylated-Interferon α-2a plus ribavirin then post-treatment viral load was assayed. Results. Hepatic expression of TLR3 was significantly increased in the non-responder group as compared to those who showed complete early virological response to the used treatment regimen. Using receiver operator characteristic (ROC) curve analysis, a cutoff level of 1.5 was set for TLR3 expression to distinguish responders from non-responders to the 12-week treatment regimen used. Conclusion. In conclusion, measuring the hepatic expression of the interferon stimulated gene, TLR3 could provide a new molecular marker for pre-treatment prediction of complete early viral clearance, thus helping in patients’ selection and optimizing treatment outcomes.

The role of immunohistochemical expression of OCT-4 and SOX2 as predictors of recurrence and progression in superficial urinary bladder carcinoma

Background Tumor recurrence and progression are two important landmarks in non-muscle-invasive bladder cancer (NMIBC) patients. Periodic cystoscopic follow-up visits of NMIBC patients are necessary after complete tumor resection as tumor recurrence occurs in nearly 70–80% of cases. Patients with a high risk for frequent tumor recurrence and disease progression may benefit from immediate adjuvant chemotherapy, immunotherapy, or even early cystoscopy. Accumulating pieces of evidence suggest that cancer stem/progenitor cells are involved in tumor relapse and therapy resistance in different solid tumors, including urinary bladder cancer (UBC), according to cancer stem cell theory. Octamer-binding transcription factor-4 (OCT-4) and sex-determining region Y-box 2 (SOX2) have been identified as two important cancer stem cell markers in various cancer tissues, not only as diagnostic markers but also as a predictor of tumor progression. In the present study, we evaluated the expression pattern of OCT-4 and SOX2 in patients with NMIBC and their role in predicting patients with a high risk for tumor recurrence and progression. Patients and methods Tumor tissue samples from 50 patients with NMIBC were collected; the pathological follow-up data of these patients were recorded; and expression status of OCT-4 and SOX2 was examined in these tissue samples by immunohistochemistry. Results Correlation between clinicopathologic parameters and follow-up results reveals that tumor size greater than 3 cm is significantly associated with tumor recurrence (MCP=0.049) and approaches level of significance with poor recurrence-free rate (MCP=0.052) based on a follow-up period of 36 months. As regards tumor progression in either histologic grade or pathologic stage, we found a significant correlation between tumor size greater than 3 cm with tumor progression (MCP=0.027) and progression-free rate (MCP=0.031). Positive OCT-4 expression was detected in 68% of cases and it was significantly associated with high tumor grade (P=0.035); this may indicate the role of OCT-4 in tumor cell dedifferentiation and promote the aggressive behavior of UBC. Positive SOX2 expression was detected in 50% of cases and was not significantly associated with tumor recurrence or tumor progression. Conclusion The present study reveals that tumor size is an important clinical risk factor for tumor recurrence and progression. Positive OCT-4 expression may be a predictive marker in NMIBC progression. More prospective studies on different stages of UBC are needed to clarify the potential role of OCT-4 in the biology and behavior of UBC. However, SOX2 has limited role in early-stage bladder cancer.

The reliability of digital image analysis and immunohistochemistry in the estimation of liver fibrosis in hepatitis C virus-related chronic hepatitis

Background Hepatitis C virus (HCV) infection is a major cause of chronic liver disease worldwide; it is characterized by liver cell injury and persistent inflammation with matrix remodeling, which leads to progressive fibrosis and eventually cirrhosis. The discovery of new screening tools for the presence of chronic liver disease and new effective therapies has created a need for a high level of reproducibility and accuracy in the evaluation of abnormal fibrosis. Several investigators have proposed the use of digital quantification as an adjunct or alternative to subjective scoring of fibrosis in liver biopsy specimens of patients with chronic HCV. Aim The aim of the present study was to measure the degree of liver fibrosis by image analysis (IA) using routine trichrome-stained sections, a cheap, available, and easily attainable stain, to evaluate the distribution and percentage of &agr;-smooth muscle actin-positive hepatic stellate cells, and to correlate both assessments with the degree of the fibrosis measured by Ishak histological scoring system. Results Thirty biopsies performed with tru-cut needle from patients with chronic HCV were analyzed. With respect to the Ishak scoring system, 14 biopsies belonged to stage 1 and 2 (seven biopsies each), 10 biopsies belonged to stage 3, three biopsies belonged to stage 4, two biopsies belonged to stage 5, and only one biopsy belonged to stage 6. A positive correlation was found between Ishak score of fibrosis and computerized IA (P⩽0.001). No correlation was detected between Ishak score of fibrosis and &agr;-smooth muscle actin immunohistochemical staining (P=0.018). From the present study, we conclude that histopathological scoring system is the gold standard for assessment of hepatic necroinflammatory activity and fibrosis stage. Conclusion We found that the fibrosis ratio of liver biopsy specimens calculated by digital IA is not always reflective of fibrosis in chronic hepatitis as indicated by subjective scoring classifications. This applies particularly to livers that are normal or show early fibrosis (stage 0–2).

The diagnostic role of fine-needle aspiration cytology combined with ultrasonography in cystic breast lesions

Background Breast cysts are among the more common reasons for referral to a breast clinic. They may be detected incidentally on screening mammography or patients may present with symptoms and signs such as mastalgia, nipple discharge, or a palpable mass. Breast ultrasound plays an established role in the evaluation of breast abnormalities, especially in women under 40 years of age. Fine-needle aspiration cytology (FNAC) has become a popular and valuable tool in the preoperative assessment of breast masses, and it shows a high accuracy, sensitivity, and specificity. The triple-diagnostic method (consisting of clinical evaluation, mammography, and FNAC) yields a precise diagnosis and reduces the risk of missed diagnosis of breast cancer to less than 1%. Aim The aim of this work was to evaluate FNAC for cystic breast lesions compared with the sonographic findings and their impact on the management policy. Patients and methods Triple assessment was carried out on 60 patients with the diagnosis of breast cysts, which included full history taking, clinical examination, ultrasonography, and FNAC. Results The most common cystic breast lesion in the present study was fibrocystic disease (46.7%), followed by abscess (20%), and the least common lesion was fat necrosis (3.3%). Regarding the radiological classification of breast cysts, simple cysts were the most common type in this study (36 cases), there were 14 cases of complicated cysts, whereas complex cysts were the least common (10 cases). Cytological examination of simple cysts was mostly bloody or inflammatory, complicated cysts were mostly inflammatory, whereas complex cysts were mostly malignant. There was a statistically significant correlation between the cyst type and the cellular content. There was no statistically significant correlation between the cyst margin and the cellular content. Regular cyst margins had a bloody and inflammatory aspirate, whereas irregular cyst margins were either inflammatory or malignant. Histopathological examination of cysts with a residual mass after aspiration showed intracystic carcinoma (six cases), abscess (two cases), fibrocystic disease (two cases), and hematoma (two cases). This correlation was statistically significant. The correlation between the cytological study and the color of the aspirate was statistically significant; malignant cells were noted mainly in bloody or greenish aspirates, whereas yellowish aspirates were always inflammatory. Conclusion The accuracy of sonography and fine-needle aspiration in detecting and allowing the correct diagnosis of cysts has been reported to be almost 100%. The use of sonography has reduced the number of biopsies performed for benign breast cysts. Aspiration should be followed by excisional biopsy when (a) blood or green fluid is aspirated or (b) the mass does not disappear completely.