Organelle Isolation for Proteomics: Mitochondria from Peripheral Blood Mononuclear Cells

Mitochondria play key roles in many cell functions including energy production, fatty acid metabolism, pyrimidine biosynthesis, calcium homeostasis, and aging. They also regulate crucial signaling cascades such as apoptosis and oxidative stress. The proteome is often used to investigate the functional correlations on protein levels. Based upon the human, genome there is estimated 2000 to 2500 associated mitochondrial proteins, however, just over 600-800 have been identified at the protein level. For this reason, mitochondria contain a great number of proteins that have yet to be identified and characterized. The identification of these proteins can help in discovery of biological process. This protocol focuses on step-by-step procedure of mitochondrial proteome extraction from peripheral blood mononuclear cell (PBMC) mitochondria. The isolation and preparation procedures described here require 6 hours approximately

Massive Uveal Relapse of Retinoblastoma Presumed to Be Choroidal Tumorous Involvement: Case Series

We report the choroidal and ciliary body invasion by retinoblastoma (RB) in a salvaged eye after complete and successful primary treatment. Case 1: A 25-month-old boy was referred due to group B RB lesions based on the International Classification of RB (ICRB; groups A–E) in the right eye (OD). His left eye (OS) was enucleated because of advanced group E RB. After 47 months of uneventful follow-up (F/U), a new lesion recurred and was treated with transpupillary thermotherapy. Four months later, a fast-growing pigmented subretinal mass was detected that was treated by brachytherapy with the apical dose of 80 Gy. Three weeks later, the lesion regressed completely, and no recurrence happened after 6 years of F/U. Case 2: A 4-month-old girl with a deletion in chromosome 13 was referred for bilateral RB. OD was enucleated because of unresponsive RB and anterior segment involvement. In OS, group B lesions had multiple recurrences after systemic chemotherapy. After a while, a single mass appeared in the nasal periphery which was controlled well with brachytherapy. Four months later, AC involvement was controlled with IAC, intravitreal, and intracameral chemotherapy, but posterior synechia and cataract appeared later. One year after the last treatment, UBM showed a ring-shaped ciliary body mass. Her parents refused enucleation again, and she received intravenous chemotherapy. Two years later, magnetic resonance imaging showed orbital and optic canal involvement with a deformed globe. In conclusion, RB recurrence can appear as local choroidal and ciliary body involvement even after a time of complete remission. The role of B-scan and UBM in early diagnosis and successful treatment is valuable

Nanofluid thin film flow and heat transfer over an unsteady stretching elastic sheet by LSM

This study is carried out on the unsteady flow and heat transfer of a nanofluid in a stretching flat plate. Least square method is implemented for solving the governing equations. It also attempts to demonstrate the accuracy of the aforementioned method compared with a numerical one, Runge-Kutta fourth order. Furthermore, the impact of some physical parameters like unsteadiness parameter (S), Prandtl number (Pr) and the nanoparticles volume fraction (ϕ) on the temperature and velocity profiles is scrutinized carefully. Accordingly, the results obtained from this study reveal that the temperature enhances by means of augmenting the nanoparticles volume fraction. At η ∈ {0, 0.5}, the velocity decreases as a result of a rise in nanoparticles volume fraction and at η ∈ {0.5, 1}, an opposite treatment takes place. Moreover, velocity distribution augments by raising the S value, however an inverse trend is observed in temperature values. Moreover, the local skin friction coefficient indicated a notable rise by increasing the S parameter as well as a steady decrease by rising ϕ. Finally, water-Alumina nanofluid demonstrated better heat transfer enhancement compared to other types of nanofluids

Study on Periplaneta americana (Blattodea: Blattidae) Fungal Infections in Hospital Sewer System, Esfahan City, Iran, 2017

Background. American cockroaches contaminated with pathogens inside hospital manholes can be one of the major problems that health care systems face. Objectives. The aim of this study was to investigate the fungal infections of American cockroaches in the Esfahan hospital sewage network. The principle goal of the study was about the roaches as a vector of fungi and other pathogens. Method. The type of study was descriptive-analytical. A total of 55 American cockroach specimens from the manhole walls of the sewerage system of 7 large hospitals were captured. Samples were taken from the surface of the body, digestive tract, and haemocoel of cockroaches. The specimens were then cultured on Sabouraud dextrose agar separately, and fungi were identified according to the macroscopic and microscopic characteristics. Results. All cockroaches collected from hospitals were infected with fungi. Among the 24 (13 infected and 11 noninfected) (44%) female cockroaches and 31 (18 infected and 13 noninfected) (56%) male cockroaches, it was identified that 40.00% was infected with Aspergillus niger, 3.64% with Rhizopus, 7.27% with Penicillium, and 5.45% with Mucor. 6 cockroaches had no yeast contamination. 17 (30.91%) cockroaches were contaminated with Candida glabrata, 23 (41.82%) cockroaches were contaminated with Candida krusei, and 22 (40%) cockroaches were contaminated with other yeast species. The results of this study showed that Candida krusei had the highest prevalence among the isolated fungi with 35.37% of the digestive system and Aspergillus niger with 70.97% of the surface of the cockroach body. Conclusion. The results emphasized the role played by cockroaches as potential pathogenic vectors in hospital environments. Therefore, suitable management is needed for controlling this insect to prevent disease transmission in hospitals

Is liver fibrosis in association with opium addiction and intravenous drug abuse among hepatitis C virus-infected patients?

Introduction: Hepatitis C virus (HCV) infection is a prevalent etiology that leads to cirrhosis. Various factors affect liver fibrosis progression. In the current study, we aimed to assess the probable role of opium consumption and intravenous drug abuse(IVDU) on liver inflammation and fibrosis. Methods: This is a case-control study conducted on 58 patients with hepatitis C virus infection in 2012. Anti-HCV antibody and quantitative HCV-RNA burden were performed for all patients. Then, they underwent a liver biopsy for the determination of inflammation grading and liver fibrosis based on the Hepatic Activity Index(HAI). Regarding inflammation grade, patients were divided into two groups of 0-4 grade as controls and 5-18 as cases. Considering the fibrosis score, patients were divided into two groups of 0-2 score as controls and 3-6 score as cases. Results: This study was conducted on HCV positive patients; among them, 74.1% were smokers, and 53.4% were opium addicts. Regarding liver inflammation grading, 52.2% of cases and 25.7% of controls were IVDU and 65.2% versus 45.7% were opium addict (P-value=0.04 and 0.145, respectively). On the other hand, regarding fibrosis score, 60% of cases versus 50% of controls were opium addicts, while 30% of cases versus 39% of controls were IVDUs (P-value>0.05). Conclusion: Contrary to the previous studies, we found no association between opium addiction with either liver inflammation or fibrosis. Based on this study, IVDU was only associated with liver inflammation, but liver fibrosis

Macular surgery using intraoperative spectral domain optical coherence tomography

Purpose: To report the use of intraoperative spectral domain optical coherence tomography (SD-OCT) for detecting anatomical changes during macular surgery. Methods: In a consecutive case series, 32 eyes of 32 patients undergoing concurrent pars plana vitrectomy and intraoperative SD-OCT for macular hole (MH), epiretinal membrane (ERM) and vitreomacular traction (VMT) were enrolled. Intraoperative changes in retinal thickness and dimensions of the macular hole were measured in patients with ERM and VMT following surgical manipulation using a hand-held SD-OCT device (iVue, Optovue Inc., Fremont, CA, USA). Results: SD-OCT images of sixteen eyes with macular hole were subjected to quantitative and qualitative analysis. All MH dimensions remained stable during consecutive stages of surgery except for MH apex diameter, which showed a significant decrease after internal limiting membrane (ILM) peeling (P=0.025). Quantitative analysis of ten patients with ERM showed a significant decrease in retinal thickness after membrane removal (P=0.018) which did not remain significant until the end of the procedure (P=0.8). In three cases, subretinal fluid was formed after ILM peeling. Quantitative analysis of five patients with VMT showed a decrease in retinal thickness during consecutive steps of the surgery, although these changes were not significant. In two cases, subretinal fluid was formed after ILM peeling. Conclusion: Intraoperative SD-OCT is a useful imaging technique which provides vitreoretinal surgeons with rapid awareness of changes in macular anatomy during surgery and may therefore result in better anatomical and visual outcomes

Quantitative measurement of vascular density and flow using optical coherence tomography angiography (OCTA) in patients with central retinal vein occlusion: Can OCTA help in distinguishing ischemic from non-ischemic type?

Abstract Background To evaluate microvascular changes and quantitative parameters in patients with central retinal vein occlusion (CRVO) by using optical coherence tomography angiography (OCTA) and finding difference between presumably ischemic and non ischemic CRVO. Methods Patients with CRVO (31) and healthy control (20) were enrolled in this observational case control study. The OCTA was done for each patient and control subject. In macular area 2 images were taken for each eye (3 × 3 mm and 8 × 8 mm). The images were analyzed at three capillary plexuses (superficial and deep retinal capillary layers and choriocapillaris layer). Results Thirty-one patients with CRVO (mean age 60.00 ± 13.72 years) and 20 healthy age/gender matched subjects (mean age 54.10 ± 12.33 years) were enrolled in this study (p = 0.095). The mean visual acuity of patients was 0.47 ± 0.54 LogMAR. Eyes with CRVO as compared with fellow eyes and control group showed significant reduction of flow in superficial (1.171 ± 0.262 vs. 1.362 ± 0.285 vs. 1.453 ± 0.105) and deep capillary plexus (1.042 ± 0.402 vs. 1.331 ± 0.315 vs. 1.526 ± 0.123) and choriocapillaris (1.206 ± 0.543 vs. 1.841 ± 0.308 vs. 1.966 ± 0.05) and vascular density in superficial (45.92 ± 4.2 vs. 50.99 ± 4.35 vs. 52.85 ± 2.99) and deep (48.03 ± 4.71 vs. 55.86 ± 3.81 vs. 58.2 ± 2.65) capillary plexuses. Some parameters (flow of both retinal capillary plexuses and parafoveal vascular density in deep plexus) showed significantly reduction in fellow eyes than control group. The parameters including flow [superficial (1.014 ± 0.264 vs. 1.279 ± 0.19) and deep (0.873 ± 0.442 vs. 1.152 ± 0.32) capillary plexuses and choriocapillaris (0.79 ± 0.327 vs. 1.424 ± 0.51)] and vascular density [superficial (44.24 ± 2.13 vs. 46.58 ± 4.13) and deep (45.28 ± 3.5 vs. 49.32 ± 3.94) capillary plexuses] were lower significantly in ischemic type than non ischemic CRVO. The most damaged parameter was flow in deep capillary plexus. The model with smallest Akaike information criterion and Bayesian information criterion was chosen as the best model. For easier calculation, we also calculated the reduced model. By choosing the threshold of 12.6, the formula [3.9 × F1S + 0.8 × F3S] can diagnose the presumably ischemic CRVO from non ischemic type with AUC of 0.84, sensitivity of 100% and specificity of 69%. (F1S: flow in the central 1 mm-radius-circle of superficial plexus and F3S: flow in the central 3 mm-radius-circle of superficial plexus). Conclusion and relevance In CRVO patients, the OCTA can accurately evaluate changes in microvascular structures. It may help in differentiation ischemic CRVO from non-ischemic CRVO