Intra-stromal injection of honey-treated keratocytes as a cell-based therapy for experimental corneal laceration.

OBJECTIVES: This study aimed to investigate the potential of honey-supplemented medium (HSM) for expanding corneal keratocytes and its transplantation in a model of corneal laceration. METHODS: Keratocytes were cultured in 1 % HSM- or 10 % fetal bovine serum (FBS)-supplemented medium for 24 h. The effect of HSM on keratocyte proliferation was evaluated using the MTT assay. The relative expression of Lum, Kera, and ALDH3A1, known markers of native keratocytes, was quantified by real-time PCR. The safety and efficacy of HSM-treated keratocyte intrastromal injection in a rabbit model of corneal laceration were also evaluated. RESULTS: The MTT assay showed that HSM treatment did not significantly affect cell viability compared to FBS-supplemented medium (84.71 ± 2.38 vs. 100.08 ± 10.92, respectively; p=0.076). Moreover, HSM-treated keratocytes had significantly increased expression of Lum, Kera, and ALDH3A1 compared to cells treated with FBS, while the expression of the proliferation biomarker Thy-1 did not significantly differ between the two treatments. Intrastromal injection of HSM-treated keratocytes in the laceration animal model was safe and uneventful, resulting in less stromal inflammation and neovascularization, and consequently, better final architecture with less residual haze compared to the group injected with FBS-treated keratocytes. CONCLUSIONS: These findings suggest that honey is a suitable supplement for keratocyte treatment and corneal cell therapy. The use of HSM may have potential applications in the treatment of corneal injuries and diseases.

Variance in the macular sublayers' volume as a diagnostic tool for primary open-angle glaucoma.

PURPOSE: To assess the variance of macular sublayers' volume in glaucoma patients compared with normal individuals. METHODS: This case-control observational study included 63 cases of primary open-angle glaucoma and 57 healthy controls. Macular volumetric scans were captured at the 6 mm ETDRS circle for each retinal sublayer using Spectralis OCT2. The studied macular sublayers included the retinal nerve fiber layer, ganglion cell layer, inner plexiform layer, inner nuclear layer, outer plexiform layer, outer nuclear layer, and outer retinal layers (external limiting membrane to the retinal pigment epithelium). Standard deviation (SD) and coefficient of variation (CoV) of macular sublayers' volume were calculated. An unpaired Student t test (or its nonparametric equivalent) was used to compare each variable between groups. The receiver operating characteristic curve (ROC) was used to investigate the discriminative ability of each parameter. RESULTS: There was no significant difference for age or sex between the groups. The SD (of all sublayers' volume) was greater in the glaucomatous eyes compared with controls (0.620 ± 0.073vs.0.524 ± 0.056  mm3, respectively; P < 0.001). The same pattern was observed for CoV (7.890 ± 0.979vs.6.128 ± 0.583; P < 0.001). The area under curves (AUCs) for SD and CoV were 0.855and0.930, respectively (P = 0.05). The best cutoff value for the CoV was 6.712. The CoV and ganglion cell layer (GCL) volume revealed similar sensitivity (80.75) at 95% specificity for diagnosing glaucoma. The CoV detected 5 patients with glaucoma who had normal GCC volume. CONCLUSION: This study showed that the macular sublayers' volume variance parameters could be viable OCT biomarkers for detecting glaucomatous changes.

The concentration of potentially toxic elements (PTEs) in drinking water from Shiraz, Iran: a health risk assessment of samples.

The existence of potentially toxic elements (PTEs) in water bodies has posed a menace to human health. Thus, water resources should be protected from PTEs, and their effect on the exposed population should be investigated. In the present investigation, the concentrations of PTEs such as lead (Pb), mercury (Hg), manganese (Mn), and iron(Fe) in the drinking water of Shiraz, Iran, were determined for the first time. In addition, hazard quotient, hazard index, cancer risk, and sensitivity analysis were applied to estimate the noncarcinogenic and carcinogenic impacts of Pb, Hg, Mn, and Fe on exposed children and adults through ingestion. The mean concentrations (µg/L) of Pb, Hg, Mn, and Fe were 0.36, 0.32, 2.28, and 8.72, respectively, in winter and 0.50, 0.20, 0.55, and 10.36, respectively, in summer. The results displayed that Fe concentration was more than the other PTEs. PTE concentrations were lower than the standard values of the Environment Protection Agency and World Health Organization. Values of the degree of contamination and heavy metal pollution index for lead, mercury, manganese, and iron were significantly low (< 1) and excellent (< 50), respectively. Based on the Spearman rank correlation analysis, positive and negative relationships were observed in the present study. The observations of the health risk assessment demonstrated that mercury, lead, iron, and manganese had an acceptable level of noncarcinogenic harmful health risk in exposed children and adults (hazard quotients < 1 and hazard index < 1). The carcinogenic risk of lead was low (< E - 06), which can be neglected. Monte Carlo simulation showed that water intake rate and mercury concentration were the most critical parameters in the hazard index for children and adults. Lead concentration was also the most crucial factor in the cancer risk analysis. The results of the present study proved that the drinking water of Shiraz is safe and healthy and can be confidently consumed by people.

Cardiac hypertrophy and fibrosis were attenuated by olive leaf extract treatment in a rat model of diabetes.

The key role of fibrosis and hypertrophy processes in developing diabetes-induced heart injury has been demonstrated. Considering the known hypoglycemic effects of olive leaf extract (OLE), we decided to investigate its potential effect and associated mechanisms on cardiac fibrosis and myocardial hypertrophy in streptozotocin (STZ)-induced diabetic rats. Eight groups were included in this study: control, diabetic, diabetic-OLEs (100, 200 and 400 mg/kg), diabetic-metformin (300 mg/kg), diabetic-valsartan (30 mg/kg), and diabetic-metformin/valsartan (300/30 mg/kg). After a treatment period of 6 weeks, echocardiography was used to assess cardiac function. Heart-to-body weight ratio (HW/BW) and fasting blood sugar (FBS) were measured. Myocardial histology was examined by Masson's trichrome staining. Gene expressions of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), ß-myosin heavy chain (ß-MHC), TGF-ß1, TGF-ß3, angiotensin II type 1 receptor (AT1), alpha-smooth muscle actin (α-SMA), and collagen were evaluated by the quantitative real-time PCR in heart tissue. A reduction in the FBS level and HW/BW ratio in the extract groups was obvious. The improvement of left ventricular dysfunction, cardiac myocytes hypertrophy, and myocardial interstitial fibrosis was also observed in treated groups. A lowering trend in the expression of all hypertrophic and fibrotic indicator genes was evident in the myocardium of OLE treated rats. Our data indicated that OLE could attenuate fibrosis and reduce myocardial hypertrophy markers, thus improving the cardiac function and structure in the STZ-induced diabetic rats. PRACTICAL APPLICATIONS: This study demonstrates that olive leaf extract in addition to lowering blood glucose levels and the heart-to-body weight ratio (HW/BW) may also improve cardiac function and reduce cardiac hypertrophy and fibrosis in cardiac tissue, which leads to inhibition of diabetic heart damage. Thus it is possible that including olive leaf extracts in the diets of individuals with diabetes may assist in lowering cardiovascular disease risk factors.

Outcomes of near confluent laser versus combined less dense laser and bevacizumab treatment of prethreshold ROP Type 1 Zone 2: a randomized controlled trial.

BACKGROUND: To evaluate the results of near confluent laser therapy versus combined less dense laser and intra vitreal bevacizumab in treatment of infants with type 1 retinopathy of prematurity (ROP) in zone II. METHODS: This is a prospective double-blinded randomized clinical trial study. Infants with Type 1 ROP in Zone 2 were randomized into case and control groups. Conventional laser therapy was executed for control group and combination of IVB and laser treatment was employed for the case group. RESULTS: Eighty-six eyes from 43 infants were analyzed in this trial. The first group included 42 eyes from 21 infants receiving a combination of laser ablation and IVB. The second group contained 44 eyes from 22 infants who received only conventional laser therapy. The combined IVB and laser ablation group demonstrated the neovascularization regression (20 out of 21 infants) one week after the procedure. In the conventional laser therapy group, this regression was found in (12 out of 22 infants) within one week after laser therapy (P = 0.001). Plus disease regression was observed in 20 (20/21) of combined treatment group and 7 infants (7/22) of conventional laser treatment group after one week. CONCLUSION: Combined less dense laser and bevacizumab treatment resulted in more rapid regression in comparison with the conventional laser treatment. TRIAL REGISTRATION: IRCT20201120049450N1, 27/12/2021.

Simultaneous removal of fluoride and nitrate from synthetic aqueous solution and groundwater by the electrochemical process using non-coated and coated anode electrodes: A human health risk study.

Co-presence of fluoride (F-) and nitrate (NO3-) in water causes numerous health complications. Thus, they should be eliminated by an appropriate method like the EC process. In this research, simultaneous removal of F- and NO3- from synthetic aqueous solution and groundwater has been considered by the EC technique under operational parameters like anode materials (un-coated (Al and Fe) and synthesized coated (Ti/TiRuSnO2 and Ti/PbO2)), cathode materials (Cu, St, and Gr), current density (12, 24, and 36 mA/cm2), inter-electrode distance (0.5, 1, and 2 cm), pH (5.5, 7, and 8.5), NaCl concentrations (0.5, 1, and 1.5 g/L), electrolysis time (15, 30, 45, 60, 90, and 120 min), NO3- concentrations (75, 150, and 225 mg/L), and F- concentrations (2, 4, 6, and 8 mg/L) for the first time in this research. The results proved that Al as non-coated anode and Cu as cathode electrodes were more effective in the co-removal of F- and NO3-. The maximum removal efficiencies of 94.19 and 95% were observed at the current density of 36 mA/cm2, 1 cm of inter-electrode distance, pH 7, 1 g/L of NaCl, and 90 min electrolysis time by Al-Cu electrode for F- (2 mg/L) and NO3- (75 mg/L), respectively. The higher efficiency of Al-Cu electrodes was due to the simultaneous occurrence of electrocoagulation, electroreduction, and electrooxidation processes. Al-Cu electrode application considerably diminished f- and NO3- concentrations in the groundwater. Health risk assessment proved that HQ of F- was significantly decreased after treatment by the Al-Cu electrode. Thus, the EC process using an appropriate and effective electrode is a promising technique for treating aqueous solutions containing F- and NO3-.

Simultaneous anaerobic decolorization/degradation of Reactive Black-5 azo dye and chromium(VI) removal by Bacillus cereus strain MS038EH followed by UV-C/H2O2 post-treatment for detoxification of biotransformed products.

The existence of synthetic dyes and heavy metals in textile wastewater is a serious problem. These compounds should be removed before discharge into the environment by an appropriate method. The present study was conducted for the characterization of efficient multi-functional strain Bacillus cereus MS038EH for the simultaneous removal of Reactive Black-5 and Chromium(VI). Maximum decolorization efficiency of 94.74% was achieved at pH 7, 35 °C, and 4% inoculum size for 900 mg/L of Reactive Black-5. Also, 94.10% efficiency was observed in the presence of 8 g/L of yeast extract as an optimum nitrogen source, while carbon sources had no significant effect on decolorization. It should be pointed out that the decolorization efficiency was decreased from 94 to 64% by increasing NaCl concentrations from 0 to 50 g/L, respectively. Bacillus cereus strain MS038EH could decolorize 94.31% of Reactive Black-5 (900 mg/L) and remove 87.31% of chromium(VI) (30 mg/L) within 36 h. Results of Gas chromatography-mass spectrometry and Fourier transform-infrared spectroscopy proved that Reactive Black-5 was cleaved into the lower molecular weight products without any azo bonds. However, the phyto-toxicity analysis showed that Reactive Black-5 was not toxic for Triticum aestivum and Maize, while biologically treated Reactive Black-5 was toxic for seeds. Therefore, ultraviolet-C/H2O2 was applied for the detoxification of biotransformed products. When ultraviolet-C/H2O2 was applied as post-treatment, the seeds were germinated completely. It is demonstrated that the application of ultraviolet-C/H2O2 after anaerobic treatment is effective for toxicity reduction of textile wastewater.

Decolorization mechanism, identification of an FMN-dependent NADH-azoreductase from a moderately halotolerant Staphylococcus sp. MEH038S, and toxicity assessment of biotransformed metabolites.

The application of halotolerant microorganisms capable of decolorizing is attractive. Decolorization mechanism, the effect of different parameters on the decolorization percentage, and toxicity analysis of Reactive Black 5 before and after decolorization were investigated in the present study. The decolorization percentage for live cells of Staphylococcus sp. strain MEH038S was more than dead cells, which demonstrated that Reactive Black 5 was decolorized through the degradation process. The results confirmed that an FMN-dependent NADH-azoreductase gene was responsible for the decolorization and then was identified as Staphylococcus sp. EFS01 azoreductase from a moderately halotolerant Staphylococcus strain for the first time. The maximal decolorization of 98.15% was observed at pH 6.5 and 35 ° C for 50 mg/L of Reactive Black 5. In addition, more than 90% decolorization was achieved with 5-40 g/L of NaCl. The results of Gas chromatography-mass spectrometry and Fourier transform infrared spectroscopy showed that Reactive Black 5 was broken to the lower molecular weight compounds without any chromophoric azo groups. Phytotoxicity and fish toxicity proved that the biotransformed metabolites of Reactive Black 5 degradation were more toxic than the original dye. The moderate halotolerant strain exhibited a remarkable decolorization capability and can be applied for textile wastewater treatment. PRACTITIONER POINTS: An azoreductase gene from a moderately halotolerant Staphylococcus was identified. More than 90% decolorization efficiency was observed under high-salt conditions. Biotransformed metabolites of RB5 degradation were identified. Toxicity analysis of biotransformed metabolites was investigated.

Effects of the secretome of human Wharton's jelly mesenchymal stem cells on the proliferation and apoptosis gene expression of the retinal pigmented epithelium.

Human retinal pigmented epithelium (RPE) can undergo an uncontrolled proliferation in some disorders such as retinal detachment associated with proliferative vitreoretinopathy (PVR). The present study was conducted to evaluate the effect of the conditioned medium secreted by human Wharton's jelly mesenchymal stem cells (WJMSCs-CM) on the proliferation and apoptosis gene expression of the RPE. WJMSCs-CM was collected from WJMSCs after two periods of 24-h and 9-h culture in serum-free medium. RPE cells were cultured in WJMSCs-CM versus serum-deprived media for 24 h. The effect of WJMSCs-CM on RPE cell proliferation was determined using the MTT assay. Relative expression of apoptotic genes (Bcl2, Bax, and IL-1B) was also assessed by real-time PCR. MTT assay demonstrated that RPE cell viability was reduced significantly in WJMSCs-CM treated RPE cells compared to those cultured in serum-deprived medium (64.23 ± 2.44 vs 100.10 ± 5.68; P = 0.006). Moreover, the expression of anti-apoptotic Bcl2 was significantly decreased in WJMSCs-CM compared to serum-deprived medium (0.52 ± 0.06 in WJMSCs-CM vs 1.02 ± 0.2 in serum-free treatment; P = 0.03), while the expression of pro-apoptotic biomarkers of Bax and IL-1B was not significantly different between the two treatments. The represented data showed that WJMSCs-CM can induce apoptosis in RPE cells in vitro through activating apoptosis pathways. This proof-of-the-concept study provides basic evidence for the possible effect of WJMSCs-CM on preventing PVR.