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Objectives: Obesity is a metabolic syndrome that leads to many chronic diseases worldwide. In this study, we investigate the antihyperlipidemic activities of chitosan nanoparticles (CH NPs) on silymarin (SIL) as a carrier in the drug delivery system that can improve some biochemical parameters and hormones in the model of hyperlipidemic rats receiving a high-fat diet (HFD)., Materials and Methods: Physicochemical characterization of silymarin-loaded chitosannanoparticles (CH-SIL NPs) was done by Fourier-transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS), and drug loading efficiency (LE). Diet-induced hyperlipidemic rats were treated with SIL (15 mg/kg/day) and CH-SIL NPs(15 mg/kg/day) for twelve weeks orally daily. The body weight loss (BW), food consumption, serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), levels of fasting blood glucose (FBG) in serum, serum insulin, cortisol, testosterone, and brain neuropeptide Y (NPY), Y1 and Y5 receptor mRNA expression were analyzed., Results: A significant reduction in BW and food consumption from 417 ± 16 g and 33 ± 1.03 in group HFD to 338 ± 10 g and 17.33 ± 1.02 in group CHS+HFD was observed, respectively. This data revealed that CH-SIL NPs improved hyperlipidemia, hyperinsulinemia, and hyperglycemia, reduced serum cortisol, and down-regulated NPY and Y1R with a significant increase in HDL and testosterone hormones compared to the control group., Conclusion: The developed Sil-loaded CH NPs were good agents for improving efficacy. It is the first report of the proposed weight loss mechanism of SIL CH NPs, thereby providing information about the anti-hyperlipidemic and antihyperglycemic effects of silymarin-loaded chitosan nanoparticles, a natural food with proper effects against metabolic disorders in case of hyperlipidemia that may lead to obesity and up-regulation of brain NPY., Competing Interests: The authors declare no conflicts of interest.

Nowadays, the Claus process is one of the most efficient procedures to recover sulfur from acid gases. In the current study, the effect of working pressure and the role of initial species (sour gas, ammonia, carbon dioxide, hydrocarbon, nitrogen, and oxygen) are analyzed using COMSOL software. The reaction occurs between acid gases, which contain 88% H 2 S, 10.5% CO 2 , 0.49% N 2 , and 1.01% CH 4 in terms of molar percentage, and pure air. A good agreement is obtained between the numerical simulation results and experimental data. According to the results, there is a direct correlation between the conversion rate of acid gases and the increase in pressure. However, this rise in reactor pressure also leads to an undesirable increase in the outlet temperature. It is also observed that reduction of hydrogen sulfide inflow decreases the sulfur monoxide production rate, which in turn significantly affects the reactor temperature and the sulfur recovery rate. The more the oxygen that enters the reactor, the more the hydrogen sulfides that change into sulfur., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Small heat shock proteins (SHSPs) are conserved proteins that participate in many cellular functions like preventing protein aggregation and stress response. However, their role in responding to nanoparticles (NPs) has not yet been explained. We used a chicken embryo model to investigate the effects of two different forms of iron oxide-NPs (IONPs) on the mRNA expression of HSPB1, HSPB5, HSPB8, and HSPB9 in cerebral tissue. Two hundred-ten fertilized eggs were randomly divided into seven groups (30 eggs/group; 10 eggs/replicate). Three groups received 100 ppm, 250 ppm, and 500 ppm of Fe 2 O 3 -NPs, respectively. Three other groups received 100 ppm, 250 ppm, and 500 ppm of Fe 3 O 4 -NPs, respectively, and one group remained untreated as a control. The NPs were given by in ovo method (0.3 ml/egg) only once on the first day of the embryonic period. Samples from cerebrums were collected on day 20 for gene expression analyses. HSPB1, HSPB5, HSPB8, and HSPB9 were all expressed in both normal and IONPs exposed cerebrums. SHSPs tested were differentially expressed in response to various concentrations of IONPs. The highest expression levels in response to Fe 2 O 3 -NPs and Fe 3 O 4 -NPs were observed for HSPB5 and HSPB9, respectively. The greatest gene expression changes due to the Fe 2 O 3 -NPs and Fe 3 O 4 -NPs exposure observed for HSPB1 and HSPB5, respectively. The results suggest a protective cellular mechanism against IONPs through SHSPs and recommend that expression profiling of SHSPs be included in the study of nanotoxicity., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Polycystic ovary syndrome (PCOS) is a common disease in women. Some plant compounds which have antioxidant properties, such as curcumin, may be useful for these patients when delivered orally or in vitro., Objective: The aim of this study was to evaluate the impact of PCOS on oocyte quality and the effect of curcumin on in vitro fertilization of oocytes., Materials and Methods: In this experimental study, Naval Medical Research Institute mice aged six to eight wk were used. Mice were divided into five experimental groups (control, experimental PCOS, curcumin 6, 12 and 24 μM). To induce experimental PCOS, estradiol valerate (100 mg/kg, IP) was injected. The total antioxidant capacity and production of malondialdehyde in ovarian tissue and blood serum were evaluated in all groups. Finally, 6, 12 and 24 μM of curcumin were added to the culture medium of the PCOS group oocytes and development in the different groups was evaluated., Results: A high percentage of oocytes for fertilization were not in good condition in terms of number and quality in the group of PCOS. The addition of curcumin to the embryo culture medium was associated with a higher percentage of fertilized oocytes, two-cells and blastocysts. This increase was significant at a concentration of 24 μM (p ≤ 0.01)., Conclusion: Given that adding curcumin seemed to improve fetal growth and prevent the harmful effects of oxygen free radicals on the culture medium, it is recommended to add a certain concentration of curcumin under normal conditions without oxidative stress., Competing Interests: The authors declare they have no conflict of interest., (Copyright © 2021 Bari et al.)

Frailty is one of the most important geriatric syndromes, which can be associated with increased risk for incident disability and hospitalization. Developing a real-time classification model of elderly frailty level could be beneficial for designing a clinical predictive assessment tool. Hence, the objective of this study was to predict the elderly frailty level utilizing the machine learning approach on skeleton data acquired from a Kinect sensor. Seven hundred and eighty-seven community elderly were recruited in this study. The Kinect data were acquired from the elderly performing different functional assessment exercises including: (1) 30-s arm curl; (2) 30-s chair sit-to-stand; (3) 2-min step; and (4) gait analysis tests. The proposed methodology was successfully validated by gender classification with accuracies up to 84 percent. Regarding frailty level evaluation and prediction, the results indicated that support vector classifier (SVC) and multi-layer perceptron (MLP) are the most successful estimators in prediction of the Fried's frailty level with median accuracies up to 97.5 percent. The high level of accuracy achieved with the proposed methodology indicates that ML modeling can identify the risk of frailty in elderly individuals based on evaluating the real-time skeletal movements using the Kinect sensor.

Lidocaine is used for epidural and spinal anesthesia in various animal species. The ideal drug for epidural and spinal anesthesia should have a long effective duration in addition to a fast onset of action, and adequate analgesia and muscle relaxation. Despite the delayed onset of action, bupivacaine provides a longer duration of anesthesia than lidocaine. The purpose of this study was to compare the onset to effect and duration of action between lidocaine and bupivacaine for spinal anesthesia in broiler chickens. Thirty-two, 8-week-old, female Ross broiler chickens were randomly divided into 4 groups of 8: 1) 2 mg/kg lidocaine (L); 2) 0.1 mg/kg bupivacaine (B0.1); 3) 0.25 mg/kg bupivacaine (B0.25); and 4) 0.5 mg/kg bupivacaine (B0.5). After aseptic preparation, a 23-gauge spinal needle was inserted into the synsacrococcygeal space of the chickens with correct needle placement confirmed by a sudden loss of resistance. Spinal anesthesia was performed with the aforementioned doses of lidocaine and bupivacaine. The respiratory rate and cloacal temperature were measured every 10 minutes in each chicken until the anesthetic effect was no longer present. The onset to effect and the duration of action were calculated for each bird based on the pinch test at predetermined time intervals. The results are demonstrated as mean ± SD. The onset of action for bupivacaine (9 ± 1.41, 4.33 ± 1.15, and 3.33 ± 1.23 minutes in B0.1, B0.25, and B0.5 groups, respectively) was significantly delayed compared with that of lidocaine (1.37 ± 0.52 minutes). The duration of action of B0.5 (54 ± 6.08 minutes) was significantly longer than that of any other group (17.87 ± 3.18, 11 ± 1.41, and 18 ± 4.36 min in L, B0.1, and B0.25 groups, respectively). The results showed that a spinal injection of 0.5 mg/kg bupivacaine produces approximately 55 minutes of spinal anesthesia in these broiler chickens, which is much longer than the 18 minutes of anesthesia provided by 2 mg/kg lidocaine. Considering the various disease conditions that affect the cloacal area of birds, one can use each of these anesthetic drugs for either short-term or long-term spinal anesthesia in chickens and possibly other avian species.

Prenatal exposure during the embryonic period has positive or adverse effect on newborn brain development. Neuroprotective activity of the hesperidin is well documented but there is no evidence for maternal exposure to hesperidin on offspring reflexive motor behaviors. So, the aim of the current study was to determine the prenatal exposure to hesperidin on reflexive motor behaviors in mice offspring. Forty pregnant female NMRI mice (8-10 weeks old) were allocated into four groups. Group 1 kept as control and groups 2-4 intraperitoneal (i.p) injected with hesperidin (0.1, 0.5, and 1 mg/kg) on days of 5, 8, 11, 14, and 17 of pregnancy. The control group injected with saline at the same days. Following delivery, 20 pups from each litter were selected and reflexive motor behaviors determined using ambulation, hind-limb foot angle, surface righting, hind-limb strength, grip strength, front-limb suspension, and negative geotaxis tests. At the end of the study serum Malondialdehyde (MDA), Superoxide dismutase (SOD), Glutathione peroxidase (GPx), and total antioxidant status (TAS) levels were determined. According to the results, maternal exposure to hesperidin (0.1, 0.5, and 1 mg/kg) increased ambulation score, front-limb suspension time, and hind-limb suspension score in mice offspring compared to the control group (p < .05). Hesperidin (0.1, 0.5, and 1 mg/kg) decreased hind-limb foot angle in mice offspring compared to the control group (p < .05). Prenatal exposure to hesperidin (0.5 and 1 mg/kg) significantly increased the surface righting and grip strength in comparison to the control group (p < .05). Hesperidin (0.1, 0.5, and 1 mg/kg) decreased MDA and increased SOD and GPx levels in mice offspring (p < .05). These results suggested hesperidin exposure during pregnancy has positive effect on reflexive motor behaviors in mice offspring may be due to its antioxidant activity., (© 2020 International Society for Developmental Neuroscience.)

Broilers are more vulnerable to high temperatures than mammals due to the feather cover, lack of sweat glands, fast growth and intensive breeding in commercial systems. Thermal stresses affect the function of various organs and change the expression profiles of hundreds of genes in the different tissues of broilers. Thermal manipulation (TM) during embryogenesis can increase heat tolerance in growing broilers. Small heat shock proteins (SHSPs) are a group of HSPs which participate in many cellular functions like response to different stressors. However, their role in the thermotolerance has not been fully elucidated. Ninety fertilized eggs were randomly divided into three groups (30 eggs/group; 10 eggs/replicate). Normal control (NC) eggs were incubated at 37.5 °C throughout the incubation period whereas heat stress (HS) and cold stress (CS) groups were kept at 41 °C and 33 °C from 15 to 17th day of incubation for 3 h each day, respectively. On day 20, samples from the cerebrums were harvested for histopathology and mRNA expression analyses of HSPB1, HSPB5, HSPB8, and HSPB9. There were no significant differences in survivability, defected embryos, hatchability, and body weight among treatments. TM had no major deleterious effects on the cerebral tissue except for mild degeneration in the HS group. HSPB1, HSPB5, HSPB8, and HSPB9 were expressed in the presence and absence of TM. All SHSP genes tested were downregulated in response to TM except for HSPB9 which was upregulated in the HS group. The highest change in gene expression due to TM observed for HSPB1. This study presents a broader understanding of mechanisms underlying response to TM in broilers. The results suggest that HSPB1, HSPB5, HSPB8, and HSPB9 are involved in thermotolerance in broilers and SHSPs could be involved in the gene expression profiling of TM. It may propose the use of nutritional supplements in the poultry industry to modulate SHSPs., (Copyright © 2020. Published by Elsevier Ltd.)

Objective: The objective of this study was to evaluate the effect of magnesium sulfate in the prevention of fetal growth restriction due to the impaired uterine blood supply in the rat model. Methods: A total number of 24 female rats were used in this study. They were mated overnight and randomly divided into control and treatment groups. After anesthesia and incising abdominal midline in day 17 of gestation, the uterine artery was occluded by an atraumatic clamp for 60 min. The rats of the control group received normal saline after surgery and the rats of treatment group received magnesium sulfate subcutaneously. The laparotomy was repeated on day 21 of gestation, and the number of alive and dead fetuses was counted in each horn. The viability of fetuses was evaluated. The weight of the placenta and fetuses and the distance between the head and tail as well as back to the abdomen of the fetuses were also measured. Samples of the amniotic fluid (AF) were collected during both surgeries for biochemical analyses of the glucose, urea, lactate, and pyruvate levels by an AutoAnalyzer. Results: Among the total fetuses in ischemic horn, only 50% survived in the control group. Dead fetuses had less body consistency and had a dark color. In contrary, only 7.6% of the fetuses in the treatment group were absorbed and 92.4% were completely healthy and developed. Parameters related to placenta weight, fetus weight, fetus length, and fetus width had significant differences and those of the treatment group were higher. Glucose and lactate levels of the AF in the treatment group were significantly lower and urea level was significantly higher than the control group in day 21 of gestation. The changes in pyruvate levels were not significant. Conclusion: In conclusion, magnesium sulfate may counteract with the effects of temporary uterine ischemia in pregnant rats and prevent intrauterine growth restriction.

Background: Paclitaxel (PTX), a chemotherapeutic agent, and monosodium glutamate (MSG) have oxidative effects on testicular tissue., Objective: In this study, the effects of MSG administration on the exacerbation of testicular tissue alterations related to PTX treatment were evaluated., Materials and Methods: MSG (30 & 60 mg/kg i.p.) was administrated to six groups (n = 8/each) of adult mice before or after PTX treatment: control, PTX-treated, MSG30 + PTX, MSG60 + PTX, PTX + MSG30, and PTX + MSG60. Following the euthanizing, the body weight measurement, pituitary-testicular axis hormonal analysis and serum lipid peroxidation index assessment was prepared, testicular histomorphometry (tubular diameter and germinal epithelium height), immunohistochemistry of p53 was completed. Microscopic indices of spermatogenesis (tubular differentiation, spermiogenesis and repopulation indices) were studied., Results: Body weight was not changed significantly. The levels of testosterone (p = 0.0001), follicle stimulating hormone (p = 0.019), and luteinizing hormone (p = 0.08) were decreased while the level of lipid peroxidation index was increased (p = 0.208) in the treated groups. The histomorphometry indices (p < 0.0001 and p = 0.001, respectively), germ cells population (p < 0.05) and microscopic indices of spermatogenesis (p = 0.001, p = 0.005, p < 0.0001, respectively) were significantly reduced in all treated groups. The administration of MSG before PTX treatment induces more changes. The most positive reaction to p53 was observed in MSG30 or 60 + PTX groups compared to other groups., Conclusion: The administration of MSG could intensify testicular tissue alterations related to PTX chemotherapy., Competing Interests: There is no conflict of interests in this study., (Copyright © 2019 Davoud Kianifard et al.)

The affiliation of the third author has been incorrectly published in the original publication of the article. The correct affiliation is provided in this erratum.

During an organism's evolution, functional adaptations help species to become better suited to their ecological niches. From the morphological aspect, these adaptations are reflected in the anatomical specializations of different organs. Specializations of the lingual organ is a critical adaptation of birds, such as the white-headed duck (Oxyura leucocephala), that enables their nutritional requirements to be met. For optimal use of the available food resources, the white-headed duck utilizes three methods of food collection, namely pecking, grazing and filter-feeding. Since this species is classified as endangered, we conducted the present study on two carcasses of the white-headed duck (death due to natural causes) employing routine histological methods, light microscopy and scanning electron microscopy. Our results show that the tongue of this bird shares some similarities and some differences with the tongue of other members of the family Anatidae. The results confirm that it is better adapted to the filter-feeding method rather than to other types of food intake. This adaptation is reflected by anatomical specializations of its lingual structures, including the stair-like outline shape, bi-sectional lingual body, a deep median sulcus, lateral conical papillae, mucus secreting glands, lack of serous secreting glands, cartilaginous skeleton and the triangular fibromuscular structure of the lingual body. The so-called triangular structure and cartilaginous skeleton are the major structures involved in the lingual motions during the filter-feeding method. The presence of the triangular structure and its connection with the cartilaginous skeleton and lingual mucosa have not previously been reported in any species of birds.

The hedgehog tongue is a tactile and taste organ which carries out various functions. Detailed functional and morphological studies are required to clearly define the relationship of the hedgehog tongue with taste, food palatability, mastication and swallowing of food, as well as the production of sounds. The aim of this study was to determine the relationship between the morphological characteristics of the European hedgehog tongue and the lifestyle of this animal, as well as to compare findings with the results of studies on other vertebrates. Gross and micro-anatomical light and scanning electron microscopy studies revealed that the hedgehog tongue could be divided in three areas, namely the apex, body and root. A keratinized stratified squamous epithelium, which was smooth on the ventral surface but bore four types of papillae on the dorsal surface, lined the tongue. Three types of these papillae were found to have gustatory functions and to express their activity in close relation with the salivary glands. These simple conical filiform papillae were situated caudally and distributed one after the other without a break. The dome-shaped fungiform papillae on the apex, with the highest distribution rate on the apex edge, were small, but those on the body and root were large. The three circular vallate papillae were arranged in a triangular shape. The foliate papillae with a few tiny projections, found in a shallow furrow, were situated between the root and the body. Most of the nerve fibers observed in different sections of the tongue tissue were of the unmyelinated type, confirming that the main task of the hedgehog tongue was its gustatory function.

Purpose: To determine the effect of folic acid (FA) on experimental testicular ischemia/reperfusion (I/R) in rats., Methods: Sixty male Wistar rats were randomly divided into 6 groups. The control group received physiologic saline orally. The sham-operated group received physiologic saline orally then exposed to midline laparotomy without clamping the IR. The I/R rats received oral gavage of the saline then subjected to 1h ischemia /24h reperfusion, period. In folic acid (2mg/kg+IR) rats received oral gavage of the FA (2mg/kg) then subjected to 1h I/24h R. groups 5-6 received FA (5 and 10 mg/kg), then subjected to 1 h I/24 h, respectively. At the end of the study, semen samples were collected for spermatozoa characteristics. The left testis was removed for histological analysis and superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GPx) measurement., Results: Spermatozoa mobility, mortality (%) significantly decreased in I/R group (P<0.05). Dose dependent increase observed on spermatozoa mobility, mortality (%) using different levels of the FA (2, 5 and 10 mg/kg) treated rat (P<0.05). Tissue MDA levels significantly increased in I/R rat (P<0.05) while FA (2, 5 and 10mg/kg) in a dose dependent manner decreased I/R-induced MDA (P<0.05). Experimental I/R significantly decreased SOD and GPx activity (P<0.05). Administration of the FA (2, 5 and 10mg/kg) significantly increased tissue SOD and GPx activity in I/R rat (P<0.05). Seminiferous tubules degenerated and loss of spermatogenesis with few spermatocytes was observed in degenerated testis tubules in I/R rat. Orally administration of the FA (5 and 10 mg/kg) improved testis characteristics with few normal seminiferous tubules and spermatocyte in seminiferous tubules in experimental I/R-induced rat., Conclusion: The treatment of folic acid had a benefit effect against ischemia-reperfusion.

Introduction: Torsion of the spermatic cord is a common urologic emergency among infants and adolescents. It requires early diagnosis and surgical intervention to prevent subfertility and infertility., Objective: The aim of this study was to investigate the effects of melatonin (MEL) and metformin (MET) co-administration on experimental testicular ischemia/reperfusion (I/R) injury in rats., Material and Methods: Fifty male Wistar rats were randomly divided into five experimental groups (n = 10), as follows. Group 1 was sham operated. In group 2, 1-hour ischemia was induced by the left testicular artery and vein clipping followed by 7 days of reperfusion. In groups 3 and 4, MEL (3 mg/kg) or MET (100 mg/kg) was administered orally for 7 days via oral gavage after ischemia, and in group 5 both agents were co-administered. At the end of trial, the left testis was removed for histological analysis and oxidative stress measurement. Histological findings in seminiferous tubule were evaluated according to Johnsen's scoring system., Results: I/R reduced superoxide dismutase (SOD) activities and testicular Johnsen's scores accompanied by an elevation in malondialdehyde (MDA) and myeloperoxidase (MPO) levels (p < 0.05). MEL and MET, and their combination restored SOD activity, tissue scores, MDA and MPO levels (p < 0.05). There was no significant difference among individual or combined treatment of these parameters (p > 0.05)., Discussion: In the present experiment, using a rat model it has been demonstrated that testicular I/R caused a significant increase in testicular injuries. This was in accordance with previous studies that have demonstrated the effect of I/R in testicular tissue. Treatment of MEL and MET had a benefit effect, but, there was no significant difference among individual or combined treatment., Conclusions: The results of the present study suggest that MEL and MET may be useful for protecting the testes from the I/R injury. However, the combined use of these agents does not further increase the protection from this damage., (Copyright © 2016 Journal of Pediatric Urology Company. Published by Elsevier Ltd. All rights reserved.)

Objective: Electroporation can be a highly efficient method for introducing the foreign genetic materials into the targeted cells for transient and/or permanent genetic modification. Considering the application of this technique as a very efficient method for drug, oligonucleotide, antibody and plasmid delivery for clinical applications and production of transgenic animals, the present study aimed to optimize the transfection efficiency of sheep testicular cells including spermatogonial stem cells (SSCs) via electroporation., Materials and Methods: This study is an experimental research conducted in Biotechnology Research Center (Avicenna Research Institute, Tehran, Iran) from September 2013 to March 2014. Following isolation and propagation of one-month lamb testicular cells (SSCs and somatic testicular cells including; Sertoli, Leydig, and myoid cells), the effect of different electroporation parameters including total voltages (280, 320, and 350 V), burst durations (10, 8, and 5 milliseconds), burst modes (single or double) and addition of dimethyl sulfoxide (DMSO) were evaluated on transfection efficiency, viability rate and mean fluorescent intensity (MFI) of sheep testicular cells., Results: The most transfection efficiency was obtained in 320 V/8 milliseconds/single burst group in transduction medium with and without DMSO. There was a significantly inverse correlation between transfection efficiency with application of both following parameters: addition of DMSO and double burst. After transfection, the highest and lowest viability rates of testicular cells were demonstrated in 320 V/8 milliseconds with transduction medium without DMSO and 350 V/5 milliseconds in medium containing DMSO. Ad- dition of DMSO to transduction medium in all groups significantly decreased the viability rate. The comparison of gene expression indicated that Sertoli and SSCs had the most fluorescence intensity in 320 V/double burst/DMSO positive. However, myoid and Leydig cells showed the maximum expression in 320 V/single burst and/or 350 V/double burst/ DMSO positive., Conclusion: We optimized the electroporation method for transfection of sheep testicular cells and recommended the application of 320 V/8 milliseconds/single pulse/DMSO negative for transduction of plasmid vector into these cells. Among testicular cells, the most external gene expression was demonstrated in SSC population.

Purpose: To investigate the protective effect of metformin on testicular ischemia/reperfusion (I/R) injury in rats., Methods: Eighteen adult male Wistar rats were randomly divided into three experimental groups (n=6), as follows: Sham, I/R, and Metformin. 1-hour ischemia was induced by the left testicular artery and vein clipping followed by 7 days of reperfusion. Metformin (100 mg/kg) was administrated orally for 7 days via oral gavage after ischemic period. At the end of trial, the left testis was removed for histological analysis and oxidative stress measurement., Results: I/R reduced superoxide dismutase (SOD) activities and testicular Johnsen's scores accompanied by an elevation in malondialdehyde (MDA) and myeloperoxidase (MPO) levels in comparison with the sham group (P < 0.05). Compared to I/R group, metformin restored testicular Johnsen's scores, SOD activity, MDA and MPO levels (P < 0.05)., Conclusion: Metformin has a protective effect against I/R injury on the testis.

Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL) in which gerbils are the reservoir host. ZCL is of great public health importance in Iran. In the current investigation, nested polymerase chain reaction (PCR) protocols were used to amplify a region of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene). The PCR assays detected L. major in three rodent species: Rhombomis opimus, Meriones lybicus and, for first time, Meriones persicus. L. major parasite was found in Natanz, Isfahan Province in the center of Iran in a focus of rural zoonotic cutaneous leishmaniasis. Four L. major infections were detected in R. opimus species, three in M. Lybicus, and two in M. persicus. All nine rodent infections of L. major were found to be the same haplotype based on the PCR detection and sequencing of parasite ITS-ribosomal DNA gene. In addition, also for the first time, the nested PCR assays detected Leishmania tropica only in one M. persicus. Allied to studies in country, the new findings mean that past conclusions about the reservoir of L. major in Iran must be treated with caution. Finding two Leishmania species in different rodent species as reservoir in Iran, therefore, careful molecular eco-epidemiological investigations will be an essential part of modeling the roles of different gerbil species in maintaining and spreading ZCL foci.