Monomeric C-Reactive Protein Localized in the Cerebral Tissue of Damaged Vascular Brain Regions Is Associated With Neuro-Inflammation and Neurodegeneration-An Immunohistochemical Study.

Monomeric C-reactive protein (mCRP) is now accepted as having a key role in modulating inflammation and in particular, has been strongly associated with atherosclerotic arterial plaque progression and instability and neuroinflammation after stroke where a build-up of the mCRP protein within the brain parenchyma appears to be connected to vascular damage, neurodegenerative pathophysiology and possibly Alzheimer's Disease (AD) and dementia. Here, using immunohistochemical analysis, we wanted to confirm mCRP localization and overall distribution within a cohort of AD patients showing evidence of previous infarction and then focus on its co-localization with inflammatory active regions in order to provide further evidence of its functional and direct impact. We showed that mCRP was particularly seen in large amounts within brain vessels of all sizes and that the immediate micro-environment surrounding these had become laden with mCRP positive cells and extra cellular matrix. This suggested possible leakage and transport into the local tissue. The mCRP-positive regions were almost always associated with neurodegenerative, damaged tissue as hallmarked by co-positivity with pTau and ß-amyloid staining. Where this occurred, cells with the morphology of neurons, macrophages and glia, as well as smaller microvessels became mCRP-positive in regions staining for the inflammatory markers CD68 (macrophage), interleukin-1 beta (IL-1ß) and nuclear factor kappa B (NFκB), showing evidence of a perpetuation of inflammation. Positive staining for mCRP was seen even in distant hypothalamic regions. In conclusion, brain injury or inflammatory neurodegenerative processes are strongly associated with mCRP localization within the tissue and given our knowledge of its biological properties, it is likely that this protein plays a direct role in promoting tissue damage and supporting progression of AD after injury.

MicroRNA155 Expression in Different Phenotypes and Genotypes of Behçet's Disease in a Sample of Egyptian Patients.

AIM: To display microRNA155 (miRNA155) expression in different entities of Behçet's disease (BD), and to find out if expression is affected in more than one of disease status than another, either phenotypically, according to HLA B51 expression, presence of family history, or patients' age. METHODS: Thirty BD patients (13 of which were HLAB51 positive) and 15 healthy subjects' samples were obtained. White blood cell miRNA155 expression in both types of samples was estimated. RESULTS: Results showed that there is a degree of relation between decrease of miRNA155 expression and different disease aspects, and also, that miRNA155 has an inverse relation with the patients' ages. CONCLUSION: MiRNA155 might be used as a measure of disease of different phenotypes, and that any manifestation of the disease can happen when the expression level decreases.

Submacular Injection of Ranibizumab as a New Surgical Treatment for Refractory Diabetic Macular Edema.

PURPOSE: In this study, we describe a new surgical technique for the treatment of refractory DME. The technique consists of vitrectomy with ILM peeling with a subretinal injection of ranibizumab. METHODS: This is a prospective interventional noncomparative study including patients with refractory DME. Included patients were subjected to the new surgical technique of pars plana vitrectomy with subretinal injection of ranibizumab. RESULTS: The study included 19 eyes with refractory macular edema, in which this novel technique was attempted. There were 10 males and 9 females. The age of the patients ranged from 17 to 67 years with a mean of 55.58 ± 13.242 years. The duration of diabetes before enrollment in the study ranged from 7 to 25 years with a mean of 16.3 years. Preoperatively, the mean CMT of the eyes ranged from 352 to 883 microns with mean ± SD of 498.58 ± 152.16 microns. Postoperatively, this improved significantly to 373.5 ± 100.3, 355.9 ± 89.8, and 365.74 ± 120.12 microns at 1, 3, and 6 months, respectively (p ≤ 0.001 for all). CONCLUSION: This novel surgical procedure of vitrectomy with ILM peeling with a subretinal injection of ranibizumab is effective in cases of refractory DME. The study has been registered in Contact ClinicalTrials.gov PRS Identifier: NCT03975088.

Evaluation of Vitrectomy with Planned Foveal Detachment as Surgical Treatment for Refractory Diabetic Macular Edema with or without Vitreomacular Interface Abnormality.

PURPOSE: To evaluate the therapeutic efficacy of subretinal BSS injections done during vitrectomy for refractory diabetic macular edema (DME) resistant to other modes of treatment including previous vitrectomy. MATERIALS AND METHODS: A prospective, interventional noncomparative case series in which cases had refractory DME with a central macular thickness (CMT) ≥ 300 µm, despite previous anti-VEGF therapy (ranibizumab or bevacizumab with shifting to aflibercept). Some cases even received intravitreal triamcinolone acetonide injection, before attempting this solution. The study included group 1, surgically naïve eyes, and group 2, cases with persistent edema despite a previous vitrectomy (7 eyes (25%)). The cases were also divided into group a, eyes with normal vitreomacular interface, and group b, with abnormal vitreomacular attachment (VMA) (6 (21.4%)). The 1ry endpoint for this study was the change in CMT after 9-12 months from surgery. The 2ry endpoints were change in BCVA, recurrence of DME, and surgical complications. RESULTS: The study included 28 eyes, 6 (21.4%) of which suffered from edema recurrence. The mean recorded CMT was 496 ± 88.7 µm and 274.1 ± 31.6 µm preoperatively and postoperatively, respectively. In all eyes, the preoperative mean BCVA in decimal form was 0.2 ± 0.11, which improved significantly to 0.45 ± 0.2. In the end, the CMT of groups 1 and 2 measured 239 µm and 170.8 µm, respectively (p = 0.019). The preoperative BCVA in groups 1 and 2 was 0.16 ± 0.07 and 0.37 ± 0.14, respectively, which improved to a mean of 0.34 ± 0.09 and 0.7 ± 0.16 postoperatively, respectively (p = 0.185). CONCLUSION: Vitrectomy with a planned foveal detachment technique was shown to be a promising solution for refractory DME cases with rapid edema resolution. CMT was shown to improve more in eyes where conventional vitrectomy was not attempted. Moreover, cases with VMA resistant to pharmacotherapy was shown to respond well to this technique. The study has been registered in Contact ClinicalTrials.gov PRS Identifier: NCT03345056.

A New Sutureless Illuminated Macular Buckle Designed for Myopic Macular Hole Retinal Detachment.

Purpose. To report the anatomic and visual results of a new sutureless illuminated macular buckle designed for patients with macular hole retinal detachment related to high myopia (MMHRD). Design. Prospective nonrandomized comparative interventional trial. Methods. Twenty myopic eyes of 20 patients (mean age, 51.4 years; range, 35-65 years) presenting with MMHRD with a posterior staphyloma, in whom the new buckle was used, were evaluated. The buckle used was assembled from a 5 mm wide sponge and a 7 mm wide silicone tire; it was fixed utilizing the sterile topical adhesive Histoacryl Blue (B Braun, TS1050044FP) which polymerizes in seconds upon being exposed to water-containing substances. The primary outcomes measured included aided visual acuity (BCVA) and optical coherence tomography (OCT) findings. The mean follow-up period was 6 months. Results. Postoperatively, the MH closure was identified by OCT in 8 (40%) eyes. The mean BCVA increased from 0.11 to 0.21 (p < 0.005). The axial length of the eyes included decreased from 30.5 mm preoperatively to 29.8 mm (p = 0.002) postoperatively. Conclusion. Preparation of the new sutureless macular buckle is simple and easy. Illumination of the terminal part of the buckle ensures proper placement. Histoacryl Blue is effective in fixing the buckle in its place for at least 6 months with no reported intra- or postoperative complications.

A Comparative Study between Vitrectomy with Internal Tamponade and a New Modified Fiber Optic Illuminated Ando Plombe for Cases of Macular Hole Retinal Detachment in Myopic Eyes.

Aim. To compare pars plana vitrectomy (PPV) with silicone tamponade or gas (Groups Ia and Ib) and a new modified Ando plombe equipped with a fiber optic light (Group II) for cases with macular hole retinal detachment (MHRD) in high myopic eyes (axial length > 26 mm). Methods. A prospective interventional randomized case series included 60 eyes (20 in each group). Successful outcome was considered if the retina was completely attached at the end of the follow-up period. Complications were identified for each group. Results. Visual acuity improved by 37.31%, 40.67%, and 49.40% in Groups Ia, Ib, and II, respectively. The success rate was 55%, 60%, and 100% in Groups Ia , Ib, and II, respectively, with a statistically significant difference between Groups Ia, Ib, and II (p < 0.001 in Ia, p: 0.002 in Ib). Complications rates were 60%, 45%, and 20% in Groups Ia, Ib, and II, respectively, with a statistically significant difference between Groups Ia and II (p: 0.01). Conclusion. Fiber optic illuminated Ando plombe allows better positioning under the macula and consequently improves the success rate of epimacular buckling in comparison to PPV with internal tamponade in MMHRD.

Control of aflatoxin production of Aspergillus flavus and Aspergillus parasiticus using RNA silencing technology by targeting aflD (nor-1) gene.

Aspergillus flavus and Aspergillus parasiticus are important pathogens of cotton, corn, peanuts and other oil-seed crops, producing toxins both in the field and during storage. We have designed three siRNA sequences (Nor-Ia, Nor-Ib, Nor-Ic) to target the mRNA sequence of the aflD gene to examine the potential for using RNA silencing technology to control aflatoxin production. Thus, the effect of siRNAs targeting of two key genes in the aflatoxin biosynthetic pathway, aflD (structural) and aflR (regulatory gene) and on aflatoxin B(1 )(AFB(1)), and aflatoxin G(1) (AFG(1)) production was examined. The study showed that Nor-Ib gave a significant decrease in aflD mRNA, aflR mRNA abundance, and AFB(1) production (98, 97 and 97% when compared to the controls) in A. flavus NRRL3357, respectively. Reduction in aflD and aflR mRNA abundance and AFB(1 )production increased with concentration of siRNA tested. There was a significant inhibition in aflD and AFB(1) production by A. flavus EGP9 and AFG(1 )production by A. parasiticus NRRL 13005. However, there was no significant decrease in AFG(1) production by A. parasiticus SSWT 2999. Changes in AFB(1) production in relation to mRNA levels of aflD showed a good correlation (R = 0.88; P = 0.00001); changes in aflR mRNA level in relation to mRNA level of aflD also showed good correlation (R = 0.82; P = 0.0001). The correlations between changes in aflR and aflD gene expression suggests a strong relationship between these structural and regulatory genes, and that aflD could be used as a target gene to develop efficient means for aflatoxin control using RNA silencing technology.

Effect of Ficus sycomorus and Azadirachta indica extracts on liver state of mice infected with Schistosoma mansoni.

The potential role of hepatoprotective and antipathological effect of Ficus sycomorus and Azadirachta indica extracts was evaluated for scavenging the reactive oxygen species (ROS) and reduced the oxidative damage and pathological changes in the liver of S. mansoni infected mice. The levels of alanine aminotransferase (ALT), asparate aminotransferase (AST) and gamma glutamyl transferase (GGT) were evaluated in the infected mice and treated orally with each plant extract 12 weeks post infection (P.I.) in a dose of 500 mg/kg of each plant extract for five consecutive days and sacrificed two weeks P.I. The infection of mice showed an elevation of ALT, AST & GGT. Treatment of mice with 70% methanol extract of each plant extract reduced significantly ALT, AST & GGT elevation. The highest reduction was with the methanolic extract of F. sycomorus (42%, 35% &44% for ALT, AST & GGT respectively). Fractionation of the methanolic extract of each plant was carried out. The effect of ethyl acetate and butanolic fractions of each plant was also evaluated. The result showed that the two fractions lowered the levels of the tested enzymes and decreased the number and size of granuloma diameters with an increased in the percentage of degenerated ova.

Differential expression of cell cycle regulators in HCV-infection and related hepatocellular carcinoma.

AIM: To investigate cell cycle proteins in chronic hepatitis C virus infection in order to analyze their role in the process of hepatocyte transformation and to characterize their prognostic properties. METHODS: Subjects of the current study included 50 cases of chronic hepatitis C (CHC) without cirrhosis, 30 cases of CHC with liver cirrhosis (LC), and 30 cases of hepatitis C-related hepatocellular carcinoma (HCC) admitted to the Department of Hepato-Gastroenterology, Theodor Bilharz Research Institute (TBRI), Giza, Egypt. Fifteen wedge liver biopsies, taken during laparoscopic cholecystectomy, were also included as normal controls. Laboratory investigations including urine and stool analysis, liver function tests and prothrombin concentration; serologic markers for viral hepatitis and ultrasonography were done for all cases of the study together with immunohistochemical analysis using primary antibodies against Cyclin D1, Cyclin E, p21, p27 and Rb/p105 proteins. RESULTS: Normal wedge liver biopsies didn't express Cyclin E or Rb/p105 immunostaining but show positive staining for Cyclin D1, p21 and p27. Cyclin D1 expressed nuclear staining that was sequentially increased from CHC to LC (P < 0.01) to HCC (P < 0.001) cases; meanwhile, Cyclin E revealed nuclear positivity only in the case of HCCs patients that was directly correlated to Rb/p105 immuno-reactivity. The expression of p21 and p27 was significantly increased in CHC and LC cases compared to normal controls and HCCs with no significant difference between well- and poorly-differentiated tumors. p21 showed only a nuclear pattern of staining, while, p27 presented with either cytoplasmic and/or nuclear reactivity in all studied cases. Correlation analysis revealed a direct relation between Cyclin D1 and p21 in CHC cases (P < 0.001), between Cyclin D1 and Cyclin E in HCCs (P < 0.01); however, an inverse relationship was detected between Cyclin D1 and p21 or p27 (P < 0.001) and between p21 and Rb/p105 (P < 0.05) in HCCs. CONCLUSION: Upregulation of Cyclin D1 in CHC plays a vital role in the development and differentiation of HCC; while, Cyclin E may be a useful marker formonitoring tumor behavior. p21 and p27 can be used as predictive markers for HCC. Furthermore, higher expression of Rb/p105 as well as inverse relation with p21 and histologic grades suggests its important role in hepatic carcinogenesis.

Possible role of cyclooxygenase-2 in schistosomal and non-schistosomal-associated bladder cancer.

BACKGROUND AND PURPOSE: Cyclooxygenase (COX) is an angiogenic factor that is strongly related to inflammatory diseases and the development of cancer and metastasis in several cancers. It is overexpressed in a variety of premalignant and malignant conditions, including urinary bladder cancer. Our aim was to investigate and compare the expression of COX-2 enzyme in patients with bladder cancer, chronic cystitis, and normal bladder tissue. The results were correlated to the classic prognostic factors, mainly tumor stage and grade, in a trial to determine the prognostic significance of COX-2 marker. MATERIALS AND METHODS: Seventy-five bladder samples were taken, including 50 cases with bladder cancer (31 were schistosomal-associated and 19 non-schistosomal-associated), 20 samples from cases with chronic cystitis (7 were nonschistosomal and 13 were schistosomal cystitis), and 5 samples from normal bladder tissue taken as control. The specimens were stained by streptavidin-biotin immunohistochemistry protocol, with COX-2 monoclonal antibody. RESULTS: Although no notable expression of COX-2 was observed in the normal bladder, it was slightly expressed in chronic cystitis especially in areas of dysplasia and squamous metaplasia, whereas there was a significant increase in COX-2 (P < .001) with moderate-to-strong granular cytoplasmic expression in all malignant histologic types. The COX-2 reactivity was higher in transitional cell carcinoma (TCC) than in squamous cell carcinoma (SqCC) (P < .01). COX-2 expression was significantly higher in schistosomal-associated TCC than in non-schistosomal-associated TCC (P < .01). There was a statistically significant positive correlation between COX-2 expression and tumor grade (P = .0052). COX-2 expression was significantly higher in grade 3 bladder TCC than in grades 1 and 2 bladder TCC (P < .05, P < .01). A correlation between COX-2 expression and progression of bladder TCC also was observed (P = .001). There was a significant difference in COX-2 expression level between the bladder TCCs at different clinical stages (P < .01). CONCLUSION: COX-2 is overexpressed in schistosomal-associated bladder cancer. COX-2 may be of significance to the development and proliferation of bladder TCC, consistent with a potential role for COX-2 inhibitors in the prevention and management of this disease.