Cadmium nanocluster as a safe nanocarrier: biodistribution in BALB/c mice and application to carry crocin to breast cancer cell lines.

Aim: Metal nanoclusters are emerging nanomaterials applicable for drug delivery. Here, the toxicity and oxidative stress induction of divalent cationic cadmium (Cd2+) was compared with a Cd in the form of nanocluster. Then, it was used for targeted drug delivery into breast cancer cell lines. Methods: Using a green chemistry route, a Cd nanocluster (Cd-NC) was synthesized based on bovine serum albumin. After characterization, its genotoxicity and oxidative stress induction were studied in both in vitro and in vivo. After that, it was conjugated with hyaluronic acid (HA). The efficiency of hyaloronized-Cd-CN (HA-Cd-NC) for loading and releasing crocin (Cro), an anticancer phytochemical, was studied. Finally, it was applied for cell death induction in a panel of breast cancer cell lines. Results: The comet assay results indicated that, unlike Cd2+ and potassium permanganate (KMnO4), no genotoxicity and oxidative stress was induced by Cd-NC in vitro. Then, the pharmacokinetics of this Cd-NC was studied in vivo. The data showed that Cd-NC has accumulated in the liver and excreted from the feces of mice. Unlike Cd2+, no toxicity and oxidative stress were induced by this Cd-NC in animal tissues. Then, the Cd-NC was targeted toward breast cancer cells by adding HA, a ligand for the CD44 cell surface receptor. After that, Cro was loaded on HA-Cd-NC and it was used for the treatment of a panel of human breast cancer cell lines with varying degrees of CD44. The half-maximal drug inhibitory concentration (IC50) of Cro was significantly decreased when it was loaded on HA-Cd-NC, especially in MDA-MB-468 with a higher degree of CD44 at the surface. These results indicate the higher toxicity of Cro toward breast cancers when carried out by HA-Cd-NC. Conclusions: The Cd-NC was completely safe and is a promising candidate for delivering anticancer drugs/phytochemicals into the targeted breast tumors.

Is it necessary to graft the void defect during open reduction and internal fixation of calcaneal fractures?

PURPOSE: There are still controversies on the effect of grafting during open reduction and internal fixation of calcaneal fractures. The aim of this study was to compare the radiological and functional outcomes in patients with or without intraoperative grafting. METHODS: In a comparative retrospective study, among 442 operatively-treated calcaneal fractures, 60 patients with unilateral closed sanders type II intraarticular calcaneal fracture who underwent ORIF via lateral extensile approach using locking anatomical plates with at least 1 year follow-up without any postoperative wound complication were enrolled. The patients were separated into 2 groups: with bone allograft and without bone allograft. The functional outcome of the patients was assessed using visual analog scale (VAS) for pain, the American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale, foot function index (FFI), and short-form (SF-36) health survey. Radiographic variables included Böhler angle, Gissane angle, calcaneal width, calcaneal height, and talar declination angle. Also, the differences (delta) of these values in comparison to the uninjured foot were calculated. RESULTS: The mean age was 39.1 ± 12.7 (range, 13-67) years with 54 males, 90.0%. No statistically significant differences were detected in age, gender, affected side, and subtypes of calcaneal fractures between the two groups (p > 0.05). The average follow-up was 25.1 (range, 12-48) months. The differences for all radiographic measurements and also, the delta values between the groups were not statistically significant, except talar declination angle which was more in cases without grafting (p = 0.007). Although the differences between the two groups regarding AOFAS ankle-hindfoot scale (p = 0.257), VAS for pain (p = 0.645), and FFI (p = 0.261) were not statistically significant; the group with bone graft experienced less pain (19.7 ± 22.0) than the other group (26.7 ± 22.8). The difference between the groups was not statistically significant (p = 0.87) according to the SF-36 questionnaire. CONCLUSIONS: Incorporating allografts into the void defects during ORIF of displaced intraarticular calcaneal fractures may not improve functional outcomes and recover postoperative radiological parameters. Therefore, routine use of allograft to fill the defects during ORIF of calcaneus may not be recommended. Of note, that these findings solely relate to the treatment of Sanders type II fractures. LEVEL OF EVIDENCE III: Comparative retrospective study.

Internal absorbed dose calculation in body organs due to injection of Rhenium-188 labeled to Mu-9 antibody.

The use of radiopharmaceuticals has gained a special place in the diagnosis and treatment of cancers and evaluation of the function of different organs of the body. In this study, the absorbed dose distribution of organs after injection of 188Re-Mu-9 has been investigated using MIRD method and MCNP-4C simulation code. The 188Re-Mu-9 labeled was injected the mouse body and the amount of 188Re-labeled accumulation was evaluated after 1, 4 and 2 4 h. Having a map of the distribution of radiopharmaceutical activity in the animal body, it is possible to convert it into a human model to obtain the internal dose received by 188Re-Mu-9 injection using the MIRD calculation method and the MCNP simulation code. According to the results of the study, the animal/human model can be acceptable method for dose estimation of antibody-based radiopharmaceuticals.

Bac to Bac System Efficiency for Preparing HPV Type 16 Virus-Like Particle Vaccine.

Today, the human papillomavirus (HPV) L1 protein is the main target in the construction of prophylactic HPV vaccines. The production of virus-like particles (VLPs) that closely resemble the natural structure of the HPV16 virus and induce high levels of virus-neutralizing antibodies in animals and humans is facilitated by the expression of HPV16-L1 protein in eukaryotic cells. The Bac-to-Bac system has been previously used to produce high levels of recombinant proteins. In this study, we utilized this expression system to generate HPV16-L1 VLPs in Spodoptra frugipedra (Sf9) insect cells. The wild-type L1 gene of papillomavirus type 16 was selected from Gene Bank and placed in bacmid structure after codon optimization using pFast Bac vector. The recombinant baculovirus containing HPV-16/L1 gene was then provided using the Bac-to-Bac system. It should be mentioned that the vector was transfected into the Sf9 cell. The cells were then lysed and the expression of L1 protein was revealed by SDS-PAGE and confirmed by Western Blot. The L1 purification was performed through Ni-NTA chromatography. The VLP formation of papillomavirus L1 protein was visualized by transmission electron microscopy. The expressed recombinant L1 was ~60 KD on SDS-PAGE which was identified in western blot by a specific anti-L1 monoclonal antibody. The electron microscopy confirmed the assembly of VLPs. Results of this study showed that the production of this protein at the industrial level can be optimized using a baculovirus/Sf9 system. The characteristics and advantages of this system are promising and it is a suitable candidate for protein synthesis.

Trastuzumab-Emtansine and Osimertinib Combination Therapy to Target HER2 Bypass Track Resistance in EGFR Mutation-Positive NSCLC.

Introduction: EGFR tyrosine kinase inhibitor improved the survival of patients with metastatic EGFR mutation-positive (EGFRm+) NSCLC. Despite high response rates, resistance develops inevitably in every patient. In up to 13%, HER2 protein overexpression is found on progression. We hypothesized that dual blockade of EGFR and HER2 by osimertinib combined with trastuzumab-emtansine (T-DM1) could reinduce tumor responses. Methods: In this multicenter, single-arm, phase 1-2 study (NCT03784599), patients with EGFRm+ NSCLC, progressing on osimertinib and HER2 overexpression were included. Patients were treated with T-DM1 3.6 mg/kg (intravenously) every 3 weeks and osimertinib 80 mg once a day. Primary end points were objective response rate (ORR) at 12 weeks and safety. Responses were assessed every 6 weeks (Response Evaluation Criteria in Solid Tumors 1.1). Sample size was calculated using Simon's two-stage minimax design (H0 = 41%, H1 > 55%, 80% power, one-sided type I error 10%: a ORR 16 of 36 was needed to proceed to 58 patients). Results: From January 2019 to April 2021, 27 patients were enrolled. ORR after 12 weeks of treatment was 4% (1 of 27). Median progression-free survival was 2.8 months (95% confidence interval: 1.4-4.6 mo). Most frequent treatment-related adverse events of any grade were fatigue, diarrhea, and nausea, among these, grade 3 in four patients. There were no grade 4 or 5 therapy-related adverse events. Conclusions: TRAEMOS (Trastuzumab-Emtansine and Osimertinib) is the first trial combining T-DM1 and osimertinib in patients with EGFRm+ NSCLC to target HER2 overexpression at osimertinib resistance. Safety profile was favorable compared with cytotoxic chemotherapy; but treatment revealed limited efficacy. Further clinical evaluation of this regimen is not warranted.

Long-Term Outcomes After Chemoradiotherapy and Surgery for Superior Sulcus Tumors.

Introduction: Superior sulcus tumors (SSTs) are uncommon, and their anatomical location can make treatment challenging. We analyzed late outcomes of patients with SST treated with concurrent chemoradiotherapy followed by surgical resection (trimodality) in a single tertiary institution. Methods: Patients with non-small cell SSTs, who underwent trimodality therapy between 2002 and 2017, were selected from a prospective institutional surgical database. Patients were uniformly staged with 18F-fluorodeoxyglucose-positron emission tomography, computed tomography scan of the chest and upper abdomen, and brain imaging. Patients undergoing resection of the lung plus chest wall were grouped as limited SST and those needing extensive resections (e.g., including the vertebral body) as extended SST. Kaplan-Meier survival analysis was performed to determine difference in survival. Multivariate Cox regression was used to identify prognostic factors. Results: A total of 123 patients were identified with a median follow-up of 4.9 years (interquartile range: 1.6-8.9 y). The 90-day postoperative mortality and morbidity (Clavien-Dindo grades III-V) were 6.5% and 21.1%, respectively. Patients with a radical resection (R0: 92.7%) had better survival (p = 0.002), as did those who had major pathologic response (73%) (p = 0.001). Ten-year overall survival (OS) and disease-free survival were 48.1% and 42.6%, respectively. There were no differences in 90-day mortality (p = 0.31) and OS (p = 0.79) between extended SST and limited SST patients. Conclusions: In patients with SST, trimodality resulted in a 10-year estimated OS and disease-free survival of 48.1% and 42.6%, respectively, which were improved after radical resection (R0) and major pathologic response. Survival for limited and extended resections was comparable, and distant relapse was the main pattern of failure. Better systemic treatments are therefore needed.

DRaW: prediction of COVID-19 antivirals by deep learning-an objection on using matrix factorization.

BACKGROUND: Due to the high resource consumption of introducing a new drug, drug repurposing plays an essential role in drug discovery. To do this, researchers examine the current drug-target interaction (DTI) to predict new interactions for the approved drugs. Matrix factorization methods have much attention and utilization in DTIs. However, they suffer from some drawbacks. METHODS: We explain why matrix factorization is not the best for DTI prediction. Then, we propose a deep learning model (DRaW) to predict DTIs without having input data leakage. We compare our model with several matrix factorization methods and a deep model on three COVID-19 datasets. In addition, to ensure the validation of DRaW, we evaluate it on benchmark datasets. Furthermore, as an external validation, we conduct a docking study on the COVID-19 recommended drugs. RESULTS: In all cases, the results confirm that DRaW outperforms matrix factorization and deep models. The docking results approve the top-ranked recommended drugs for COVID-19. CONCLUSIONS: In this paper, we show that it may not be the best choice to use matrix factorization in the DTI prediction. Matrix factorization methods suffer from some intrinsic issues, e.g., sparsity in the domain of bioinformatics applications and fixed-unchanged size of the matrix-related paradigm. Therefore, we propose an alternative method (DRaW) that uses feature vectors rather than matrix factorization and demonstrates better performance than other famous methods on three COVID-19 and four benchmark datasets.

Preparation of porous PCL-PEG-PCL scaffolds using supercritical carbon dioxide.

In this study, the Supercritical Carbon Dioxide (scCO2) gas foaming procedure was used in the preparation of scaffolds containing the model drug dexamethasone (DXMT). The method used did not include an organic solvent thus making it a safe method. The ring-opening polymerization of PCL-PEG-PCL (PCEC) triblock was conducted using an organocatalyst [1,8 diazabicyclo [5.4.0] undec-7-ene (DBU)]. After mixing 5.0 g of DXMT with 50.0 g of PCEC, hydraulic pressure was applied to compress the mixed powder into disc-like tablets. The tablet-like scaffold of the triblock containing DXMT was inserted into a scCO2 gas-foaming device. The peak porosity percentage of the synthesized triblock was found to be 55.58 %. Pressure, temperature, soaking time and the time required to depressurize were recorded as 198 bar, 50 °C, 2.0 h, and 28 min respectively. After treatment with scCO2, the scaffolds experienced an almost full release of DXMT in vitro after 30 days (83.74 ± 1.54 % vs 52.24 ± 2.03 % before scCO2 treatment). In conclusion, the results proved that the scCO2 gas foaming procedure could be employed for constructing modifiable PCEC scaffolds with plausible porosity and structural and morphological features which can manipulate drug release.

The effect of cochlear implant insertion technique on post-operative neural response telemetry and impedance in paediatric patients.

OBJECTIVE: This study aimed to compare neural response telemetry and impedance between the round window and cochleostomy approaches for cochlear implantation. METHODS: In this case-control study, 64 patients aged less than 3.5 years underwent cochlear implantation via the round window or cochleostomy approach. Post-operative neural response telemetry and impedance were measured. RESULTS: The impedance measurements at electrodes 1, 11 and 22 showed no significant differences between the two groups three months after implantation (p = 0.90, p = 0.08 and p = 0.37, respectively). Similar results were observed six months after implantation (p = 0.71, p = 0.65 and p = 0.70, respectively). There was no significant difference in neural response telemetry between the two groups after three months. The neural response telemetry of electrode 1 in the cochleostomy group (171.26 ± 19.81 µV) was significantly higher in comparison with that of electrode 1 in the round window group (161.97 ± 12.71 µV) after six months (p = 0.03). The neural response telemetry values for electrodes 11 and 22 did not show any significant difference after six months (p = 0.14 and p = 0.48, respectively). CONCLUSION: Both approaches provide equal stimulation of the cochlear nerve and impedance.

The first report of tinea nigra from Iran.

Hortaea werneckii causes Tinea nigra, a rare superficial mycosis. It has not been reported in Iran yet. We report a case of an Iranian boy resident of Amol (Mazandaran, Iran) that developed brown macules on his left palm. Direct microscopic examination and culture confirmed the diagnosis of Tinea nigra.

Impact of smoking status on the relative efficacy of the EGFR TKI/angiogenesis inhibitor combination therapy in advanced NSCLC-a systematic review and meta-analysis.

BACKGROUND: The ETOP 10-16 BOOSTER trial failed to demonstrate a progression-free survival (PFS) benefit for adding bevacizumab to osimertinib in second line. An exploratory subgroup analysis, however, suggested a PFS benefit of the combination in patients with a smoking history and prompted us to do this study. METHODS: A systematic review and meta-analysis to evaluate the differential effect of smoking status on the benefit of adding an angiogenesis inhibitor to epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor therapy was carried out. All relevant randomized controlled trials appearing in main oncology congresses or in PubMed as of 1 November 2021 were used according to the Preferred Reporting Items for Systematic Review and Meta-Analyses statement. Primarily PFS according to smoking status, and secondarily overall survival (OS) were of interest. Pooled and interaction hazard ratios (HRs) were estimated by fixed or random effects models, depending on the detected degree of heterogeneity. Bias was assessed using the revised Cochrane tool for randomized controlled trials (RoB 2). RESULTS: Information by smoking was available for 1291 patients for PFS (seven studies) and 678 patients for OS (four studies). The risk of bias was low for all studies. Combination treatment significantly prolonged PFS for smokers [n = 502, HR = 0.55, 95% confidence interval (CI): 0.44-0.69] but not for nonsmokers (n = 789, HR = 0.92, 95% CI: 0.66-1.27; treatment-by-smoking interaction P = 0.02). Similarly, a significant OS benefit was found for smokers (n = 271, HR = 0.66, 95% CI: 0.47-0.93) but not for nonsmokers (n = 407, HR = 1.07, 95% CI: 0.82-1.42; treatment-by-smoking interaction P = 0.03). CONCLUSION: In advanced EGFR-non-small-cell lung cancer patients, the addition of an angiogenesis inhibitor to EGFR-tyrosine kinase inhibitor therapy provides a statistically significant PFS and OS benefit in smokers, but not in non-smokers. The biological basis for this observation should be pursued and could determine whether this might be due to a specific co-mutational pattern produced by tobacco exposure.

A theoretical study on spontaneous dipole orientation in ice structures.

Spontaneous dipole orientation is studied for a set of simulated porous ASW ice films on a substrate held at temperatures ranging from 10 K to 140 K. It is found that the water dipoles in the films obtained at the lower temperatures are oriented such that a negative electric field with a magnitude of 108-109 V m-1 is obtained. The magnitude of the field increases approximately linearly with height above the substrate, akin to experimental observations, although the magnitude of our field increases faster. A strong temperature dependence of the surface potential resulting from the spontelectric field is found, where the surface potential decreases when the substrate temperature increases. The surface potential finally becomes close to zero for temperatures around and above 110 K.

Neoscytalidium dimidiatum as onychomycosis causative agent in an Iranian patient: a case report and literature review.

A 38-year-old healthy male presented to our medical mycology center with whitish opaque discoloration of the right toenail. He reported a history of some sand scratches subsequent to walking barefoot on the beach two years ago and wearing hard safety shoes for a period of two years. On clinical examination, onycholysis, onychodystrophy, and apparent thickening of the ungual bed in the left big toe were found. The microscopic examination of nail clippings using 15% potassium hydroxide (KOH/) revealed the presence of septate pigmented hyphae. The fungus was identified as Neoscytalidium dimidiatum based on the cultural characteristics, the arrangement of arthroconidia on lactophenol cotton blue (LPCB) staining, blocky-brown pigmented hyphae on serum physiology mounts, and sequencing. Susceptibility of the isolated fungi to amphotericin B, itraconazole, voriconazole, and terbinafine was tested using the standard broth microdilution M38-A2 method developed by the Clinical and Laboratory Standards Institute (CLSI). The minimum inhibitory concentrations (MICs) of the four antifungal drugs used in this study were: amphotericin B: 1 mg/L, itraconazole: 2 mg/L, voriconazole: 0.25 mg/L, and terbinafine: 1 mg/L. The patient underwent terbinafine and clobetasol topical treatments for 6 months.

Dosimetric verification of IMRT and 3D conformal treatment delivery using EPID.

PURPOSE: Electronic portal imaging devices (EPIDs) could potentially be useful for either in-vivo or pre-treatment dosimetric verification of external beam radiation therapy. The accuracy of EPID for dosimetric purposes is highly dependent on the specific method used for the determination of dose-response characteristics. The aim of this study was to develop a simple and time-saving EPID back-projection dosimetry algorithm for 2D dose verification in 3D conformal and intensity-modulated beams. METHODS: The procedure of dose reconstruction includes a first calibration step using ionization chamber measurements to convert the Electronic Portal Image (EPI) pixel values into an absorbed dose in water. Subsequently, several corrections were applied to the Portal Dose Images (PDIs) for the effect of field size, attenuator thickness, scattering radiation, beam hardening and EPID off-axis response. Furthermore, to consider tissue inhomogeneity for accurate dose reconstruction, the patient's water equivalent path length (WEPL) was calculated using a range of digitally reconstructed radiographs (DRRs) obtained at various thicknesses by Plastimatch software. The EPID-derived dose maps accuracy was assessed by comparing with the treatment planning system (TPS) calculated dose in the prostate region of Alderson phantom irradiated with 3D conformal and intensity-modulated beams. RESULTS: The gamma analysis for the dose plane showed agreements of 96.95% and 93.5% for 3D conformal and IMRT fields, respectively, with 3%/3 mm acceptance criteria. CONCLUSION: The presented algorithm can provide accurate absolute 2D dose maps for clinical use in the context of 3DCRT or IMRT Quality Assurance (QA) programs.

A randomised phase II study of osimertinib and bevacizumab versus osimertinib alone as second-line targeted treatment in advanced NSCLC with confirmed EGFR and acquired T790M mutations: the European Thoracic Oncology Platform (ETOP 10-16) BOOSTER trial.

BACKGROUND: While osimertinib, a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) is the standard treatment in patients with advanced non-small-cell lung cancer (NSCLC) with sensitising EGFR and acquired T790M mutations, progression inevitably occurs. The angiogenic pathway is implicated in EGFR TKI resistance. PATIENTS AND METHODS: BOOSTER is an open-label randomised phase II trial investigating the efficacy and safety of combined osimertinib 80 mg daily and bevacizumab 15 mg/kg every 3 weeks, versus osimertinib alone, in patients with EGFR-mutant advanced NSCLC and acquired T790M mutations after failure on previous EGFR TKI therapy. Primary endpoint was investigator-assessed progression-free survival (PFS). Secondary endpoints were overall survival (OS), objective response rate (ORR) and adverse events (AEs). RESULTS: Between May 2017 and February 2019, 155 patients were randomised (combination: 78; osimertinib: 77). At data cut-off of 22 February 2021, median follow-up was 33.8 months [interquartile range (IQR): 26.5-37.6 months] and 129 (83.2%) PFS events were reported in the intention-to-treat population. There was no difference in median PFS between the combination [15.4 months; 95% confidence interval (CI) 9.2-18.0 months] and osimertinib arm (12.3 months; 95% CI 6.2-17.2 months; stratified log-rank P = 0.83), [hazard ratio (HR) = 0.96; 95% CI 0.68-1.37]. Median OS was 24.0 months (95% CI 17.8-32.1 months) in the combination arm and 24.3 months (95% CI 16.9-37.0 months) in the osimertinib arm (stratified log-rank P = 0.91), (HR = 1.03; 95% CI 0.67-1.56). Exploratory analysis revealed a significant interaction of smoking history with treatment for PFS (adjusted P = 0.0052) with a HR of 0.52 (95% CI 0.30-0.90) for smokers, and 1.47 (95% CI 0.92-2.33) for never smokers. ORR was 55% in both arms and the median time to treatment failure was significantly shorter in the combination than in the osimertinib arm, 8.2 months versus 10.8 months, respectively (P = 0.0074). Safety of osimertinib and bevacizumab was consistent with previous reports with grade ≥3 treatment-related AEs (TRAEs) reported in 47% and 18% of patients on combination and osimertinib alone, respectively. CONCLUSIONS: No difference in PFS was observed between osimertinib plus bevacizumab and osimertinib alone. Grade ≥3 TRAEs were more common in patients on combination.

The in silico mechanism of hVKOR interaction with acetaminophen and its metabolite, as well as N-acetyl cysteine: caution on application in COVID-19 patients.

Acetaminophen and N-acetyl cysteine (NAC) are being used as supportive care in patients suffering from coronavirus disease 2019 (COVID-19). The coagulopathy and cerebral hemorrhage have been recently reported in these patients. Prolonged acetaminophen use increases the international normalized ratio (INR) and the risk of bleeding among patients taking anti-coagulants. Inhibition of vitamin K epoxide reductase (VKOR) by acetaminophen and NAC in chronic applications has been reported, however, detailed knowledge of the molecular mechanism and binding sites are not clear. Herein, we built the homology model of human VKOR (hVKOR) using ITASSER server, confirmed, and applied it for docking analysis of its interaction with acetaminophen and its metabolite, N-acetyl-p-benzoquinone imine (NAPQI), and NAC. We also calculated the lipophilicity and predicted the blood-brain-barrier (BBB) permeation of NAPQI by Swiss ADME. Our analysis showed that NAPQI and NAC, but not acetaminophen, bind strongly to the similar sites in hVKOR via both hydrogen and van der Waals bonding; particularly with Cys135. Thus, it interrupted the vitamin K reducing electron transfer pathway. Further, molecular dynamic (MD) simulation study revealed that the interactions of the ligands with hVKOR are stable. In conclusion, our analysis shed a light on the molecular mechanism of acetaminophen-induced coagulopathy previously reported in some clinical cases with chronic acetaminophen use. Furthermore, considering the anti-coagulopathy of NAPQI and NAC but not acetaminophen, the BBB permeation potency of these agents, and the risk of coagulopathy in COVID-19, we suggest a regular prothrombin time (PT) and INR monitoring of these patients taking acetaminophen and/or NAC.Communicated by Ramaswamy H. Sarma.

Characterization and Comparison of Mesenchymal Stem Cell-Derived Exosome Isolation Methods using Culture Supernatant.

Exosomes are extracellular endosomal nanoparticles, which are formed under complex processes during the formation of multivesicular bodies. They are also achieved from conditioned media of a variety of cell types, especially mesenchymal stem cells (MSCs). Exosomes can modulate intracellular physiological actions via signaling molecules on the surface or secretion of components to the extracellular spaces. Furthermore, they are potentially used as crucial agents for cell-free therapy; however, their isolation and characterization can be challenging. In the current study, two methods of exosome isolation have been characterized and compared using a culture media of adipose-derived mesenchymal stem cells, namely ultracentrifugation and a commercial kit; moreover, the efficiency of these two methods was highlighted in this study. Two different isolation methods of exosomes from MSCs were used to compare the efficiency of exosomes. For both isolation methods, transmission electron microscopy, dynamic light scattering (DLS), and bicinchoninic acid (BCA) assay have been performed. The electron microscopy and DLS indicated the presence of exosomes. Moreover, the kit and ultracentrifugation isolates contained approximately comparable amounts of protein measured by the BCA. Overall, the two isolation methods had similar performances. Although ultracentrifugation is used as a gold standard for exosome isolation, the commercial kit has some advantages and can be applied alternatively according to its cost-effectiveness and time-saving properties.

The Effects of Nano and Micro Particle Size of Zinc Oxide on Performance, Fertility, Hatchability, and Egg Quality Characteristics in Laying Japanese Quail.

The current trial was conducted by using the 288 Japanese quails to evaluate the effects of a zinc-deficient diet supplemented with nano and micro zinc oxide on performance, fertility, hatchability, and egg quality characteristics. In this experiment, birds were randomly allocated to 9 dietary treatments includes diets supplemented with nano or micro particles of zinc oxide (amount of ZnO supplement for each treatment) to supply 49, 74, 99, and 124 mg zinc per kilogram of diet in a factorial arrangement (2 × 4) and a control non supplemented diet (24 mg/kg) with four replicates of eight birds (six females and two males) in each pen. Birds were fed the experimental diets from 47 to 75 days of age and had free access to water and feed during the experimental period. Results showed that Zn supplementation, regardless of particle size, improved the eggshell thickness (P < 0.01). A significant (P < 0.05) interaction was observed between zinc level and ZnO particle size for Shell breaking strength. Quails fed diets supplement with ZnO showed significantly higher egg weight and eggshell surface (P < 0.05) as compared with birds fed a non-supplemented control diet. Results obtained here showed that supplementation of nano ZnO enhanced fertility considerably. Application of non-linear quadratic models showed that the maximum egg production percentage was achieved when 67 or 72 mg/kg of dietary zinc was supplied from nano and micro ZnO, respectively. This result indicated that nano ZnO could reduce the zinc requirement in laying Japanese quail.

A comparison effects of l-citrulline and l-arginine against cyclosporine-induced blood pressure and biochemical changes in the rats.

OBJECTIVE: The use of cyclosporine A (CsA) is associated with different adverse effects including hypertension and nephrotoxicity. The present study aimed to compare the inhibitory effects of l-arginine &l-citrulline on CsA-induced blood pressure and biochemical changes in the serum of rats. METHODS: Thirty-six rats were divided into 6 groups received daily: (1) 1ml distilled water, (2) 200mg/kg l-citrulline IP, (3) 25mg/kg CsA SC, (4) CsA+l-citrulline with the same dose of the former groups, (5) 200mg/kg l-arginine IP and (6) l-arginie+CsA with the same doses of group 4 for 7 days. RESULTS: The changes in the blood pressure, heart rate, creatinine, BUN, glucose and C-reactive protein (CRP) of the serum were determined in the treated animals. Significant (p<0.001) increase was shown in the blood pressure and heart rate of CsA treated rats compared to the control group. There were also a significant (p<0.05) increase in the creatinine, BUN and glucose, but a decrease in the CRP value in the CsA-treated group. However, l-citrulline significantly (p<0.001) inhibited the changes in the blood pressure and heart rate in CsA-treated as well as it was able to reduce blood pressure in non-treated group significantly (p<0.01). l-citrulline also inhibited the increased levels of BUN and creatinine induced by CsA, while, l-arginine was able to prevent the increased blood pressure and creatinine occurs after administration of CsA. CONCLUSIONS: These findings suggest that the l-citrulline is more efficient than l-arginine against the adverse effects induced by cyclosporine.

Subtilisin Gene Activity in Dermatophytes: A study on the Presence of the Subtilisin Gene in Trichophyton verrucosum and Microsporum gypseum in Clinical and Nonclinical Samples in Tehran, Iran.

The keratinolytic activities of dermatophyte species are accompanied by the secretion of enzymes, such as serine proteases, which are coded by the Subtilisin (SUB) genes. This study aimed to determine the presence of the SUB genes in the clinical and nonclinical samples of Trichophyton verrucosum and Microsporum gypseum. Isolation was carried out by direct and laboratory examination. Following that, for the determination of the presence of the SUB gene, polymerase chain reaction with specific primers was conducted. The frequencies of the SUB gene were observed in almost 66% of the isolates. Statistical analysis showed a significant relationship between the presence of the SUB gene and the samples collected from human, animals, and soil (p ˂0.005). The current investigation has been the first study of the presence/absence of the SUB gene in the clinical and nonclinical isolates of T. verrucosum and M. gypseum in Iran which may be a new step to perform further studies.