Protective effect of dental pulp stem cells' conditioned medium against cisplatin-induced testicular damage in rats.

Cisplatin is a highly effective chemotherapy drug used to treat most solid tumors. However, one of its side effects is testicular toxicity, which can lead to fertility abnormalities. This study investigated the effectiveness of dental pulp mesenchymal stem cells conditioned medium (DPSC-CM) on cisplatin-induced testicular toxicity. In this study, 36 eight-week-old male Wistar rats were randomly divided into three groups equally (n = 12). Group 1 control "CTR", which received normal saline (0.5 ml) intraperitoneally (i.p), group 2 "Cis" which received an intraperitoneal dose of cisplatin (7 mg/kg), and group 3 "Cis+CM" which received an i.p injection of DPSC-CM (0.5 mg/kg) after cisplatin injection. Biochemical, histomorphometric, and histopathological studies were performed on the testis. Our results exhibited that cis administration led to a decline in total body weight, testis weight, diameter, and volume. A decrease in testosterone and IL-6 serum levels, as well as a decrease in IL-6 and TNFα levels, the activity of catalase and SOD enzymes, and an increase in MDA in testicular tissue were detected. Testicular tissue damage was associated with a significant decrease in tube diameter, germinal epithelium height, number of spermatogonia and Sertoli cells, along with a noticeable increase in basement membrane thickness, and perivascular fibrosis. DMSC-CM improved all the mentioned parameters. Taken together, our results demonstrated that DMSC-CM due to its antioxidant and anti-inflammatory properties, could be effective in reversing cisplatin-induced testicular toxicity.

Clinical Correlation of Altered Molecular Signatures in Epileptic Human Hippocampus and Amygdala.

Widespread alterations in the expression of various genes could contribute to the pathogenesis of epilepsy. The expression levels of various genes, including major inhibitory and excitatory receptors, ion channels, cell type-specific markers, and excitatory amino acid transporters, were assessed and compared between the human epileptic hippocampus and amygdala, and findings from autopsy controls. Moreover, the potential correlation between molecular alterations in epileptic brain tissues and the clinical characteristics of patients undergoing epilepsy surgery was evaluated. Our findings revealed significant and complex changes in the expression of several key regulatory genes in both the hippocampus and amygdala of patients with intractable epilepsy. The expression changes in various genes differed considerably between the epileptic hippocampus and amygdala. Different correlation patterns were observed between changes in gene expression and clinical characteristics, depending on whether the patients were considered as a whole or were subdivided. Altered molecular signatures in different groups of epileptic patients, defined within a given category, could be viewed as diagnostic biomarkers. Distinct patterns of molecular changes that distinguish these groups from each other appear to be associated with epilepsy-specific functional consequences.

Evaluation of sperm chromatin/DNA integrity, morphology, and Catsper expression on diabetic C57BL/6 mice.

BACKGROUND: Diabetes is associated with reproductive impairment on the male reproductive system and causes complications such as decreased libido, fertility, spermatogenesis, sperm motility, and morphology. High levels of blood sugar may affect sperm quality and reduce the potential for male fertility. Increased levels of sperm DNA damage is often associated with reduced count and motility or abnormal morphology. METHODS: This experimental study was conducted in Mashhad University of Medical Sciences. In this work, 40 mice (C57BL/6) were divided randomly into 4 groups: 1) Control, 2) Diabetic, 3) Diabetic + Insulin, and 4) Sham. After 35 days, the right epididymis of all specimens was used for Real-Time PCR and left epididymis for evaluation of sperm parameters using Aniline blue, Toluidine blue, Papanicolaou, and immunohistochemical study. Also, testes were applied for immunohistochemical, TUNEL studies, and biochemical assay. RESULTS: Results of this study showed that chromatin integrity, morphology, cation channels of sperm (Catsper) expression, and biochemical factors level were significantly changed in diabetic mice in comparison to other groups (P<0.05) and treatment with insulin improved these parameters. CONCLUSION: Our findings showed that the sperm parameters such as DNA integrity, morphology, and Catsper expression change in diabetic mice.

The protective effect of co-administration of platelet-rich plasma (PRP) and pentoxifylline (PTX) on cyclophosphamide-induced premature ovarian failure in mature and immature rats.

Cyclophosphamide (CP), as an antineoplastic agent, causes premature ovarian failure (POF) due to ovarian toxicity and subsequent infertility in women. Platelet-rich plasma (PRP) has accumulated significant attention in regenerative medicine. Pentoxifylline (PTX) as a methylxanthine derivative has been shown to have antioxidant and antiapoptotic properties. The aim of this study was to evaluate the protective effect of PRP and PTX on CP-induced POF. Fifty mature and immature female rats were assigned into five groups: control, CP (75 mg/kg, intraperitoneal [ip] on days 1 and 10 to induce POF), CP + PRP (200 µl, ip, half an hour after CP injection on day 1 and 10), CP + PTX (50 mg/kg, orally, half an hour after CP injection daily for 21 day), and CP + PRP + PTX. At the end of experiments on day 21, measurement of body weight, ovarian parameters (ovarian volume, follicular granulosa cell layers diameter, oocyte diameter, and the number of granulosa cells), measurement of ovarian hormone in sera for estradiol (E2), and anti-Mullerian hormone (AMH), as well as biochemical assessment were performed.The results showed that CP significantly reduced the ovarian parameters, E2, AMH, superoxide dismutase (SOD) activity and increased Malondialdehyde (MDA) levels compared to the control group (p < 0.001). Our results also indicated that all histomorphometric parameters and biochemical markers in CP-induced POF, were preserved close to normal by PRP and PTX treatments in both mature and immature rats (p < 0.001). Therefore, it is concluded that the co-administration of PRP and PTX can protect the ovary from CP-induced POF.

Differentiation of spermatogonial stem cells by soft agar three-dimensional culture system.

Since proliferation and differentiation of spermatogonial stem cells (SSCs) in culture system provide successful transplantation in this study, culture of human SSCs was compared to SACS (soft agar culture system), gelatin and control groups. The cells were isolated from seminiferous tubules of non-azoospermia patients (NOA) and cultured in DMEM for 3 weeks. The presence of SSCs in culture system was confirmed by immunocytochemistry of GFR-α1 and ITGα6 antibodies. The proliferated cells were cultured in three mentioned groups in the presence of retinoic acid and Sertoli cells conditioned medium for another 2 weeks. The number of colonies in the SACS group was significantly higher than two other groups. Before 2 weeks of culture, only Oct4 expression was observed in testicular cells (2.32 ± 0.25). After 2 weeks, the expression of Oct4 in the gelatin group was higher than that of the SACS group on day 7. The maximum expression of Stra8 was observed in SACS and gelatin groups after 7 days, but its expression was significantly decreased after 14 days of culture (p < .05). The expression of Scp3 and Acrosin genes were higher after 14 days in the SACS group compared to other groups. SACS has positive effects on proliferation and differentiation of hSSCs.

Cell injury and receptor expression in the epileptic human amygdala.

Neuropathological findings in the amygdala obtained from patients with mesial temporal lobe epilepsy (MTLE) indicate varying degrees of histopathological alterations, such as neuronal loss and gliosis. The mechanisms underlying cellular damage in the amygdala of patients with MTLE have not been fully elucidated. In the present study, we assess cellular damage, determine the receptor expression of major inhibitory and excitatory neurotransmitters, and evaluate the correlation between the expression of various receptors and cell damage in the basolateral complex and the centromedial areas in the amygdala specimens resected during brain surgery on 30 patients with medically intractable MTLE. Our data reveal an increased rate of cell damage and apoptosis as well as decreased expression levels of several GABAergic receptor subunits (GABAARα1, GABAARß3, and GABABR1) and GAD65 in the amygdalae obtained during epilepsy surgery compared to autopsy specimens. Analyses of the expression of glutamate excitatory receptor subunits (NR1, NR2B, mGluR1α, GluR1, and GluR2) reveal no significant differences between the epileptic amygdalae and autopsy control tissues. Furthermore, the increased occurrence of apoptotic cells in the amygdala is negatively correlated with the reduced expression of the studied GABAergic receptor subunits and GAD65 but is not correlated with the expression of excitatory receptors. The present data point to the importance of GABAergic neurotransmission in seizure-induced cell injury in the amygdala of patients with MTLE and suggest several GABA receptor subunits as potential druggable target structures to control epilepsy and its comorbid disorders, such as anxiety.

Assessment of sperm morphology, chromatin integrity, and catSper genes expression in hypothyroid mice.

There is an evident relationship between the fertilizing capacity of sperm and the normal morphology, quality chromatin, and motility of sperm. It is well known that thyroid hormones are the important regulators of testicular function. A correlation was found between the hypothyroidism and sperm damages. The present study was conducted to investigate the effects of hypothyroidism on sperm morphology, chromatin quality, and motility. For this purpose, 20 male mice were divided into the control and the hypothyroid groups that received 0.05% 6-n-propyl-2-thiouracil (PTU) for 35 days. Sperm morphology with Papanicolaou staining and sperm chromatin quality with both Aniline Blue (AB) and Toluidine blue (TB) staining were assessed. Besides, immunohistochemistry and real-time PCR were performed to evaluate the changes of cation sperm channel (CatSper) genes. A significant increase in the sperm chromatin condensation was found in the hypothyroid mice compared to the control mice (p < 0.05). Furthermore, a significant decrease was observed in the morphology of normal sperm in hypothyroid mice compared to the controls (p < 0.05). The results showed that Hypothyroidism could downregulate the expression of CatSper genes. Immunohistochemical data confirmed the real time-PCR results. Furthermore, the results showed that hypothyroidism could adversely affect sperm morphology, sperm chromatin condensation, and CatSper gene expression in mice and these abnormalities may be related to the excessive production of reactive oxygen species (ROS) in a hypothyroid state.

Resveratrol promotes the arcuate nucleus architecture remodeling to produce more anorexigenic neurons in high-fat-diet-fed mice.

OBJECTIVE: Adult hypothalamic neurogenesis has been considered a central regulator of energy balance. Resveratrol (RSV), a natural polyphenol, influences the body fat mass and reduces the amount of adipose tissue. The present study was designed to evaluate the effect of RSV on dynamic of hypothalamic neurons in a diet-induced obesity model of mice. METHODS: Apoptosis, neurogenesis, the expression of the main trophic factors, and the fate of newborn cells were evaluated in the hypothalamus of adult male C57 BL/6 J mice fed a normal diet, a high-fat (HF) diet, or an HF diet supplemented with 400 mg/kg RSV (HF + RSV) for 6 wk. RESULTS: The HF diet caused an increase in neuronal apoptosis in the hypothalamus, which coincided with an increase in the number of newborn cells in the arcuate nucleus, suggesting that compensatory mechanisms developed to overcome deleterious effects of the HF diet. Addition of RSV to the HF diet enhanced the production of newborn cells in all studied regions of the hypothalamus. These changes were paralleled by enhancement of the expression of ciliary neurotrophic factor. Interestingly, a considerable proportion of newborn cells expressed neuropeptide Y in the arcuate nucleus of the HF group, and conversely, most of them differentiated to proopiomelanocortin neurons in HF + RSV mice. CONCLUSIONS: Diets rich in fat changed hypothalamic neuronal balance toward orexigenic versus anorexigenic neurons. Administration of RSV to the HF diet reversed this balance toward generation of anorexigenic neurons. These data point to the potential for RSV in regulation of body weight, possibly via modulation of hypothalamic neurogenesis.

Protective effects of ascorbic acid and garlic extract against lead-induced apoptosis in developing rat hippocampus.

Lead exposure has negative effects on developing nervous system and induces apoptosis in newly generated neurons. Natural antioxidants (i.e. Ascorbic acid and Garlic) might protect against lead-induced neuronal cell damage. The aim of the present study was to investigate the protective effects of Ascorbic acid and Garlic administration during pregnancy and lactation on lead-induced apoptosis in rat developing hippocampus. Timed pregnant Wistar rats were administrated with Lead (1500 ppm) via drinking water (Pb group) or lead plus Ascorbic acid (Pb + AA Group, 500 mg/kg, IP), or lead plus Garlic Extract (Pb + G Group, 1 ml garlic juice/100 g BW, via Gavage) from early gestation (GD 0) until postnatal day 50 (PN 50). At the end of experiments, the pups' brains were carefully dissected. To identify neuronal death, the brain sections were stained with TUNEL assay. Mean of blood and brain lead levels increased significantly in Pb group comparing to other studied groups (P < 0.01). There was significant reduction in blood and brain lead level in Pb + AA and Pb + G groups when compared to those of Pb group (P < 0.01). The mean number of TUNEL positive cells in the CA1, CA3, and DG was significantly lower in the groups treated by either Ascorbic acid or Garlic (P < 0.05). Administration of Ascorbic acid and Garlic during pregnancy and lactation protect against lead-induced neuronal cell apoptosis in the hippocampus of rat pups partially via the reduction of Pb concentration in the blood and in the brain.

Preparing neural stem/progenitor cells in PuraMatrix hydrogel for transplantation after brain injury in rats: A comparative methodological study.

Cultivation of neural stem/progenitor cells (NS/PCs) in PuraMatrix (PM) hydrogel is an option for stem cell transplantation. The efficacy of a novel method for placing adult rat NS/PCs in PM (injection method) was compared to encapsulation and surface plating approaches. In addition, the efficacy of injection method for transplantation of autologous NS/PCs was studied in a rat model of brain injury. NS/PCs were obtained from the subventricular zone (SVZ) and cultivated without (control) or with scaffold (three-dimensional cultures; 3D). The effect of different approaches on survival, proliferation, and differentiation of NS/PCs were investigated. In in vivo study, brain injury was induced 45 days after NS/PCs were harvested from the SVZ and phosphate buffered saline, PM, NS/PCs, or PM+NS/PCs were injected into the brain lesion. There was an increase in cell viability and proliferation after injection and surface plating of NS/PCs compared to encapsulation and neural differentiation markers were expressed seven days after culturing the cells. Using injection method, transplantation of NS/PCs cultured in PM resulted in significant reduction of lesion volume, improvement of neurological deficits, and enhancement of surviving cells. In addition, the transplanted cells could differentiate in to neurons, astrocytes, or oligodendrocytes. Our results indicate that the injection and surface plating methods enhanced cell survival and proliferation of NS/PCs and suggest the injection method as a promising approach for transplantation of NS/PCs in brain injury.

Hinged Capsulotomy--Does it Decrease Floaters After Yttrium Aluminum Garnet Laser Capsulotomy?

OBJECTIVES: The objective was to compare conventional circular yttrium aluminum garnet (YAG) laser capsulotomy with hinged capsulotomy to manage posterior capsular opacification (PCO). MATERIALS AND METHODS: This prospective, randomized clinical trial enrolled pseudophakic patients with visually significant posterior capsule opacification. Patients were randomized to undergo posterior YAG laser capsulotomy with either conventional circular technique or a new technique with an inferior hinge. At 1-month postoperatively, patients were asked if they had any annoying floaters and the responses were compared between groups. P < 0.05 was considered statistically significant. RESULTS: A total of 83 patients were enrolled. Forty-three patients underwent hinged posterior YAG capsulotomy and 40 patients underwent routine circular capsulotomy. At 1-month postoperatively, there was a statistically significant decrease in annoying floaters in the group that underwent circular capsulotomy (P = 0.02). There was no statistically significant association in the total energy delivered (P = 0.4) or the number of spots (P = 0.2) and patient perception of annoying floaters. CONCLUSION: Hinged YAG capsulotomy was effective at decreasing the rate of floaters in patients with PCO.

The neuroprotective effect of a triazine derivative in an Alzheimer's rat model.

UNLABELLED: Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder. It is characterized by formation of amyloid plaques and neurofibrillary tangles in the brain, degeneration of the cholinergic neurons and neural cell death. This study was aimed to investigate the effect of a triazine derivative, C16H12Cl2N3S, on learning in an Alzheimer's rat model. Animals were divided into seven groups; each group contained seven animals. CONTROL GROUP: animals received no surgery and treatment; saline group: animals received normal saline after recovery; sham group: animals received 10% DMSO after recovery; STZ group (Alzheimer's model): animals received streptozotocin (STZ) in four and six days after recovery; T5, T10 and T15 groups: animals were treated with triazine derivative, C16H12Cl2N3S, at doses of 5, 10 and 15 µM, respectively. All drugs were injected intracerebroventricular. The spatial learning and histological assessment were performed in all groups. Animals in STZ group had more deficits in spatial learning than the control group in Morris water maze. C16H12Cl2N3S improved spatial learning significantly compared to STZ group. The CA1 pyramidal layer thicknesses in STZ group were reduced significantly compared to control group. C16H12Cl2N3S increased the CA1 pyramidal layer thickness in T15 group compared to STZ group. Current findings suggest C16H12Cl2N3S may have a protective effect on learning deficit and hippocampal structure in AD.

Can Valeriana officinalis root extract prevent early postoperative cognitive dysfunction after CABG surgery? A randomized, double-blind, placebo-controlled trial.

RATIONALE: We hypothesized that valerian root might prevent cognitive dysfunction in coronary artery bypass graft (CABG) surgery patients through stimulating serotonin receptors and anti-inflammatory activity. OBJECTIVES: The aim of this study was to evaluate the effect of Valeriana officinalis root extract on prevention of early postoperative cognitive dysfunction after on-pump CABG surgery. METHODS: In a randomized, double-blind, placebo-controlled trial, 61 patients, aged between 30 and 70 years, scheduled for elective CABG surgery using cardiopulmonary bypass (CPB), were recruited into the study. Patients were randomly divided into two groups who received either one valerian capsule containing 530 mg of valerian root extract (1,060 mg/daily) or placebo capsule each 12 h for 8 weeks, respectively. For all patients, cognitive brain function was evaluated before the surgery and at 10-day and 2-month follow-up by Mini Mental State Examination (MMSE) test. RESULTS: Mean MMSE score decreased from 27.03 ± 2.02 in the preoperative period to 26.52 ± 1.82 at the 10th day and then increased to 27.45 ± 1.36 at the 60th day in the valerian group. Conversely, its variation was reduced significantly after 60 days in the placebo group, 27.37 ± 1.87 at the baseline to 24 ± 1.91 at the 10th day, and consequently slightly increased to 24.83 ± 1.66 at the 60th day. Valerian prophylaxis reduced odds of cognitive dysfunction compared to placebo group (OR = 0.108, 95 % CI 0.022-0.545). CONCLUSION: We concluded that, based on this study, the cognitive state of patients in the valerian group was better than that in the placebo group after CABG; therefore, it seems that the use of V. officinalis root extract may prevent early postoperative cognitive dysfunction after on-pump CABG surgery.

Six year Trend in Cataract Surgical Techniques in Iran.

PURPOSE: To determine the cataract surgery techniques performed in Iran from 2000 to 2005. MATERIALS AND METHODS: This study was part of the Iranian Cataract Surgery Survey (ICSS) which was a retrospective cross-sectional study. All major ocular surgery units and 10% of randomly selected minor units throughout Iran were included. Excluding the 2 week Iranian New Year holiday, 1 week per season between 2000 and 2005 (a total of 24 weeks) was selected for each center, and data on all cataract surgeries performed during these weeks were collected by reviewing patient records. The ANOVA repeated measure test was performed to determine longitudinal changes with a P<0.05 denoting statistical significance. RESULTS: Phacoemulsification with intraocular lens (IOL) implantation has become the surgical method of choice in Iran, increasing from less than 7% in 2000 to 57% in 2005 (P<0.0001). Extracapsular cataract extraction showed a reverse trend compared to phacoemulsification, decreasing from greater than 91% in 2000 to 41% in 2005 (P<0.0001). Intracapsular cataract extraction and lensectomy were rarely performed without significant changes over time (P>0.05). CONCLUSION: Phacoemulsification with IOL implantation has become the preferred cataract surgery method in Iran during recent years.

Ocular manifestations of transconjunctival heroin abuse: a case report of an unusual route of substance abuse.

PURPOSE: The purpose of this study was to present a patient with atypical corneoconjunctival lesions as a result of transconjunctival heroin abuse. METHODS: The authors conducted a prospective 8-month follow up of a 16-year-old girl who presented with bilateral atypical corneoconjunctival lesions. RESULTS: After a period of close observation, running diagnostic tests, and conservative treatment, the nature of the disease was suspected. Further discussion with the patient's family and her final confession revealed that she was using heroin through the conjunctiva. CONCLUSION: Substance abuse should be considered in the list of differential diagnoses in patients with an unusual presentation of ocular disease.

Five year cataract surgical rate in Iran.

PURPOSE: To determine the number of cataract surgeries per million population per year or the cataract surgical rate (CSR) in Iran for every year from 2000 to 2005 as part of the objectives of the Vision 2020 initiative by the World Health Organization (WHO) to eliminate cataract blindness. METHODS: This study is part of the Iranian Cataract Surgery Survey, which was conducted as a retrospective cross-sectional study. All major cataract surgery centers and 10% of randomly selected minor centers throughout the nation were approached. Excluding the 2-week Iranian New Year holiday, 1 week per season between 2000 and 2005 (a total of 24 weeks) was selected for each center, and data concerning all cataract surgeries performed during these weeks were recorded by reviewing records of patients operated for cataracts. RESULTS: A total of 13,409 surgical records from 2000 to 2005 were evaluated, and CSR was calculated for each year based on the population data obtained from the Statistical Center of Iran. CSR showed an increasing trend in Iran, rising from 526 (95% confidence interval: 352 to 700) in year 2000 to 1331 (95% confidence interval: 1084 to 1577) in 2005. CONCLUSIONS: CSR has significantly increased in Iran in recent years but is still below the target suggested by WHO. Considering the age structure of the young population of Iran, the present CSR may suffice. However, it is necessary to implement plans to increase CSR and obviate obstacles to eliminating cataract blindness.