L-carnitine protects human retinal pigment epithelial cells from oxidative damage

PURPOSE: To determine the efficacy of L-carnitine (LC) against oxidative changes in human retinal pigment epithelium (RPE) cells. METHODS: The RPE cells from human donor eyes were cultured in Hams F-10 medium. The effect of LC on H2O2-induced morphologic changes in the RPE cells was analyzed by light microscopy. Reduction in cell death after the impact of LC treatment on H2O2-treated cells was analyzed by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays. In addition, the effect of H2O2 on the activity of RPE-antioxidant enzymes, glutathione (GSH) and superoxide dismutase (SOD), and LC-induced protection was also determined. RESULTS: LC protected the RPE cells by inhibiting the peroxide-induced cytopathic effect from 50% to 10%. Nuclear condensation observed in 40% of the H2O2-treated cells decreased to 20% after LC treatment. The MTT assays demonstrated that 100 microM oxidant caused appreciable cell death, which was reduced by LC treatment; however, 100% protection was not achieved. Significant peroxide-induced cell death was seen within 5 hr of H2O2 treatment, and a quantifiable reduction was observed after LC treatment for a similar time period. The change in the antioxidant potential of the RPE induced by oxidative stress was restored by LC treatment, as demonstrated by an increase in GSH and SOD activities. CONCLUSIONS: LC is capable of protecting the RPE cells from H2O2-induced oxidative damage, implying that micronutrients can have a positive effect and can play an important role in the treatment of oxidation-induced ocular disorders. Further studies are needed to understand the mechanism of LC-induced protection to the RPE cells.     

Detection of Mycobacterium leprae in ocular tissues by histopathology and real-time polymerase chain reaction

AIM: To report detection of leprosy in ocular tissue by histopathology and its confirmation by genetic analysis. METHODS: Excised tissue from a clinically-suspected ocular leprosy patient was processed and analyzed histopathologically. The DNA from the paraffin-embedded tissue was extracted, an 85 A-C intergenic region of Mycobacterium leprae was amplified using specific primers and analyzed by conventional as well as real-time polymerase chain reaction (RT-PCR). RESULTS: With periodic acid-Schiff-hematoxylin (PAS-H) staining the specimen showed presence of a thin fibrinous layer of inflammatory cells. The majority of the tissue was fibrovascular with extensive infiltration by histiocytes having reticulated cytoplasm. Modified PAS-H and acid-fast staining (AFS) showed the presence of several acid-fast organisms within the cytoplasm of histiocytes and mast cells. Conventional PCR showed a 250-bp DNA from excised conjunctival tissue, which was in agreement with the positive controls for M. leprae. Through RT-PCR, it was calculated that the suspected tissue had 44.68 pg of M. leprae DNA, which is 8937.06 genome copies of M. leprae. CONCLUSIONS: Presence of inflammatory cells and AFS bacilli in tissue presented a typical picture of leprosy. M. leprae DNA can be detected using RT-PCR in ocular tissues when acid-fast bacteria are seen in histopathological sections. And when the diagnosis of leprosy is inconclusive and acid-fast bacteria are seen, RT-PCR for M. leprae DNA could be used as a rapid confirmatory test to identify the presence of M. leprae and, therefore, the diagnosis of leprosy.     

临床表现似视网膜母细胞瘤的脉络膜黑色素细胞瘤1例(英文)

我们报道罕见的脉络膜黑色素细胞瘤1例,以视网膜母细胞瘤之诊断行眼球内容物剜除术。患儿14mo,以左眼外斜视约10mo主诉就诊。眼科检查及影像学检查显示视网膜脱离并钙化病灶。患儿诊断为视网膜母细胞瘤,患眼行眼球内容物剜除术。组织病理学检查显示肿瘤细胞体积大,多面体型,核小,胞浆含大量黑色素颗粒,提示脉络膜及睫状体黑素细胞瘤之诊断。以往有脉络膜及睫状体黑色素细胞瘤表现类似与恶性黑素瘤的报道,但是,黑色素细胞瘤表现为钙化病灶及视网膜脱离,被误诊为视网膜母细胞瘤,此病例罕见,以往未见报道。     

Inhibition of RPE lysosomal and antioxidant activity by the age pigment lipofuscin.

PURPOSE: To determine whether lipofuscin is detrimental to lysosomal and antioxidant function in cultured human retinal pigment epithelial (RPE) cells. METHODS: Isolated lipofuscin granules were fed to confluent RPE cultures and the cells maintained in basal medium for 7 days. Parallel cultures were established that did not receive lipofuscin. Cultures were either exposed to visible light (390-550 nm) at an irradiance of 2.8 mW/cm(2) or maintained in the dark at 37 degrees C for up to 24 hours. Cells were subsequently assessed for cell viability, lysosomal enzyme activity, and antioxidant capacity. RESULTS: There was no loss of cell viability during the first 3 hours of light exposure, whereas a 10% loss of viability was observed in lipofuscin-fed cultures after 6 hours' exposure to light. Activities of acid phosphatase, N-acetyl-beta-glucuronidase, and cathepsin D were decreased by up to 50% in lipofuscin-fed cells exposed to light compared with either unfed cells or cells maintained in the dark. There was also a decrease in the antioxidant potential of RPE cells. Catalase and superoxide dismutase activities decreased by up to 60% and glutathione levels by 28% in light-exposed lipofuscin-fed cells compared with unfed cells or cells maintained in the dark. CONCLUSIONS: Lipofuscin has the capacity to reduce the efficacy of the lysosomal and antioxidant systems in RPE cells that may play an important role in retinal ageing and the development of age-related macular degeneration.     

The effect of jaw relaxation on pain anxiety during burn dressings: Randomised clinical trial

Aim

The purpose of this randomised clinical trial (RCT) was to determine the effect of jaw relaxation on pain anxiety related to dressing changes in burn injuries.

Introduction

Patients hospitalised with burns experience high levels of anticipatory anxiety during dressing changes, which cannot be completely managed by anxiolytic drugs. Nurses as members of the burn care team contribute to pain management by using relaxation techniques as one of the most frequently used approaches to pain anxiety management. However, there is not enough information about the effects of these techniques on pain anxiety of patients with burns. The aim of this study was to determine the effect of jaw relaxation on pain anxiety related to dressing changes in burn injuries.

Methods

It was a randomised clinical trial with a control group. A total of 100 patients hospitalised in Shahid Motahari Burn Centre affiliated with Tehran University of Medical Sciences were recruited by convenience sampling and were randomly assigned to either experimental or control groups using minimisation. With institutional approval and written consent, the experimental group practiced jaw relaxation for 20 min before entering the dressing room. Data were collected by the Burn Specific Pain Anxiety Scale (BSPAS) during July–December 2009 and analysed using Statistical Package for the Social Sciences (SPSS)-PC (17).

Results

An independent t-test showed no significant difference between mean pain anxiety scores in the experimental and control group before intervention (p = 0.787). A dependent t-test showed significantly less pain anxiety after intervention (before dressing) in the experimental group (p < 0.05). Moreover, the independent t-test showed that the post-dressing pain anxiety of the experimental group was less than the control group (p < 0.05). However, the dependent t-test showed no significant difference between before and after dressing pain anxiety (after intervention) in the experimental group (p = 0.303).

Conclusion

Nurses can independently decrease the pain anxiety of patients with burns and its subsequent physical and psychological burden by teaching the simple and inexpensive technique of jaw relaxation. Further research is needed to study the effect of this technique on pain anxiety of patients suffering from other painful procedures.     

Alpha-lipoic acid effect on leptin and adiponectin concentrations: a systematic review and meta-analysis of randomized controlled trials

New evidence suggests that dysregulation of adipocytokines caused by excess adiposity plays an important role in the pathogenesis of various obesity comorbidities. Our aim in this meta-analysis was to determine the effect of alpha-lipoic acid (ALA) supplementation on serum levels of leptin and adiponectin. We searched Scopus, PubMed, Google Scholar, and ISI Web of Science from inception up to July 2019. Mean difference for leptin and adiponectin were calculated by subtracting the change from baseline in each study group. Summary estimates for the overall effect of ALA on serum leptin and adiponectin concentrations were calculated using random effects model. Results were presented as weighted mean difference (WMD) and their 95% confidence intervals (CI). Between-study heterogeneity was examined using the I2 statistics. Eight studies were included in systematic review and seven studies in meta-analysis. The overall effect suggested a significant decrement in serum leptin concentrations (WMD = − 3.63; 95% CI, − 5.63, − 1.64 μg/ml; I2 = 80.7%) and a significant increase in serum levels of adiponectin (WMD = 1.98 μg/ml; 95% CI, 0.92, 3.04; I2 = 95.7%). Subgroup analyses based on age showed a significant reduction in leptin levels only in younger adults, and subgroup analysis based on duration indicated in studies with a duration of more than 8 weeks adiponectin levels increased significantly and leptin levels decreased significantly. Our results revealed ALA decreased leptin and increased adiponectin especially in studies lasted more than 8 weeks. We still need more studies with different ALA dose, intervention duration, and separately on male and female.     

The epigenetic alterations of human sperm cells caused by heroin use disorder

The molecular mechanisms of drug use on sexual health are largely unknown. We investigated, the relationship between heroin use disorder and epigenetic factors influencing histone acetylation in sperm cells. The volunteers included twenty-four 20- to 50-year-old men with a normal spermogram who did not consume any drugs and twenty-four age- to BMI-matched men who consume only the drug heroin for more than last four months. HDAC1 and HDAC11 mRNA expression levels in spermatozoa and miR-34c-5p and miR-125b-5p expression levels in seminal plasma were measured. The heroin-user group showed significantly increased white blood cell counts and decreased sperm motility and survival rates (8.61 ± 1.73, 21.50 ± 3.11, 69.90 ± 4.69 respectively) as compared to the control group (1.49 ± 0.32, 38.82 ± 3.05, 87.50 ± 0.99 respectively) (p ≤ .001). An increase in DNA fragmentation index (DFI) (heroin-user group: 41.93 ± 6.59% and control group: 10.14 ± 1.43%, p = .003), a change in frequency of HDAC1 (heroin-user group: 1.69 ± 0.55 and control group: 0.45 ± 0.14, p = .045) and HDAC11 (heroin-user group: 0.29 ± 0.13 and control group: 2.36 ± 0.76, p = .019) in spermatozoa and a significant decrease in seminal miR-125b-5p abundance (heroin-user group: 0.37 ± 0.11 and control group: 1.59 ± 0.47, p = .028) were reported in heroin consumers. Heroin use can lead to male infertility by causing leukocytospermia, asthenozoospermia, DFI elevation in sperm cells and alterations in seminal RNA profile.     

Randomized controlled trial of the foot reflexology on pain and anxiety severity during dressing change in burn patients

BackgroundOne of the most important problems in burn patients was pain, especially in dressing changes. This pain can lead to anxiety in the patient. The aim of this study was to determine the effect of foot reflexology on pain and anxiety severity in burn patients.MethodsThis study was a randomized controlled trial, in which 66 patients with burn injuries referred to Vali-e-asr Hospital, Arak, Iran participated. After obtaining written consent, patients were enrolled to study according to inclusion criteria and then, divided into intervention (n = 33) and control (n = 33) groups using simple random allocation. In the intervention group, in addition to standard care, reflexology was performed for one week on Saturday, Monday and Wednesday (three times in a week). The intervention was done one hour before dressing change in a separate room for 30 min. The control group received only standard care during this time (both intervention and control groups were the same in the type of treatment, and reflexology was considered as an extra care in the intervention group). Severity of pain and anxiety in both groups was measured using visual analog scale twice a day (5?10 min before dressing change and 5?10 min after dressing change) for six days. SPSS software ver. 15 was used for statistical analysis. Mean and standard deviation were used for quantitative variables and qualitative variables were reported as frequency and percentage. Data were analyzed using Chi?square, Mann–Whitney, Fisher’s exact tests, and paired t-test. The Kolmogorov–Smirnov test was used to check the normality of data.ResultsThe results showed no significant difference in severity of pain (p = 0.25) and anxiety (p = 0.37) between the two groups on the first day, before the intervention. In the following days, the results showed no significant difference between the two groups in the second and third treatments after intervention . However, the mean pain scores showed a significant difference between the two groups in the forth (p = 0.005), fifth (p = 0.001), and sixth (p = 0.001) days after intervention. Anxiety scores also showed a significant difference between the two groups on the fourth (p = 0.01), fifth (p = 0.001), and sixth (p = 0.001) days.ConclusionsOur results showed foot reflexology is an appropriate and safe intervention for management of pain and anxiety of burn patients. Therefore, it can be used as a complementary method alongside other methods.     

RNAi mediated gene silencing of ITPA using a targeted nanocarrier: Apoptosis induction in SKBR3 cancer cells

A pure nucleotide pool is required for high‐fidelity DNA replication and prevention of carcinogenesis in living cells. Human inosine triphosphatase (ITPase), encoded by the ITPA gene, plays a critical role in maintaining the purity of the cellular nucleotide pool by excluding nucleotides that enhance mutagenesis. ITPase is a nucleoside triphosphate pyrophosphatase that hydrolyzes the non‐canonical nucleotides inosine triphosphate (ITP) and xanthine triphosphate (XTP). The monophosphate products of ITPase reactions are subsequently excluded from the nucleotide pool and the improper substitution of ITP and XTP into DNA and RNA is prevented. Previous studies show that deficiency in ITPA can suppress cellular growth and enhance DNA instability. In this study, we evaluated the influence of effective ITPA down‐regulation on the induction of apoptosis in a human cancer cell line using folate‐single wall nanotubes (SWNT) as a targeted nanocarrier. We assessed whether SWNT enhances IPTA‐siRNA transfection efficiency in cancer cells using folate as a homing device. Since folate receptor is considerably overexpressed in cancer cells, conjugation of SWNTs to folate could enhance their cancer‐specific penetrance. We found that nanocarrier mediated ITPA‐siRNA transfection into SKBR3 cells caused significant reduction of ITPA mRNA expression level and complete down‐regulation of the ITPase protein product. The silencing of ITPA led to promotion of apoptosis in SWNT‐treated SKBR3 cancer cells.