Cytotoxic and Radiosensitizing Effects of Folic Acid-Conjugated Gold Nanoparticles and Doxorubicin on Colorectal Cancer Cells.

Purpose: Radiotherapy is one of the most important therapeutic options used to treat cancers. Radiation effects can be improved using nanoparticles and chemotherapeutic drugs as radiosensitizing agents. The aim of the present study was to evaluate the effects of folic acid-conjugated gold nanoparticles (GNP-F) in combination with doxorubicin (DOX) and x-Ray irradiation in colorectal cancer (CRC) cell line (HT-29). Methods: The cell viability assay (WST-1) was performed to study the cytotoxic effects of different concentrations of DOX and GNP-F after 24 and 48 hours treatments. Then, the effects of the GNP-F, X-Ray irradiation, and DOX drug in single and combined treatments were examined after 24 and 48 hours treatment with effective doses. Likewise, the caspase 3 gene expression ratio and the caspase 3 activity were assessed after 48 h treatment. Moreover, the malondialdehyde (MDA) level was determined in treated and untreated cells. Results: When GNP-F (at a concentration of 70 µM) was combined with X-ray irradiation (2 Gy) and DOX drug, induced more cytotoxic effects compared to the control group. The results of cell viability assay showed that GNP-F + X-Ray in combination with a low concentration of DOX (0.25 × IC50) enhanced the cytotoxic effects of cells compared to related single treatments. Caspase 3 gene expression ratio and caspase 3 activity increased in double and triple combination treatments in comparison with the single groups. Moreover, the MDA level increased in triple combination compared to the single treatments. Conclusion: Our findings confirmed the potential anti-cancer effects of the GNP-F and DOX in combination with X-Ray irradiation in CRC cells.

In Vitro and in Vivo Scolicidal Activities of Holothuria leucospilota Extract and CeO2 Nanoparticles against Hydatid Cyst.

BACKGROUND: We aimed to investigate the scolicidal effects of Holothuria leucospilota extract and CeO2 nanoparticles against protoscoleces of hydatid cysts in-vitro and in-vivo. METHODS: Hydatid cysts were collected from, Urmia slaughterhouses between years 2016-2017 and the hydatid fluid aspirated from the fertile cysts. Various concentration of H. leucospilota extract, CeO2 NPs and combination of CeO2-NPs/H. leucospilota were used for 10-60 min to evaluate the viability of protoscoleces by 0.1% eosin method. CASPASE -3 activity measured for assessment of cell apoptosis in treated protoscoleces. BALB/c mice were infected intraperitoneally with 2000 viable protoscoleces and treated daily for 4 wk by intragastrical inoculation with H. leucospilota, CeO2 NPs, combination of CeO2 NPs/H. leucospilota and Albendazole. Cyst development was macroscopically analyzed. RESULTS: H. leucospilota extract and combination of CeO2 NPs/H. leucospilota have potent scolicidal activity at concentration of 20 mg/ml and 15 mg/ml after 60 min treatment. Maximum caspase-3 activity was observed when protoscoleces expose with H. leucospilota and combination H. leucospilota & CeO2 NPs. After treatment of cyst infected mice with extract and CeO2 NPs, combination of CeO2 NPs/H leucospilota and albendazole, a significant decrease in number of cysts, size and volume of cyst (P<0.05) was observed. CONCLUSION: This result shows an antihydatic and scolicidal effects of H. leucospilota extract and CeO2 NPs.

Nandrolone administration with or without strenuous exercise increases cardiac fatal genes overexpression, calcium/calmodulin-dependent protein kinaseiiδ, and monoamine oxidase activities and enhances blood pressure in adult wistar rats.

Misuse of anabolic androgenic steroids (AAS) increases prevalence of cardiovascular abnormalities in athletes, and the underlying molecular mechanism involved in those abnormalities continues to be investigated. The aim of this study was to investigate the effect of chronic nandrolone exposure on alpha and beta-myosin heavy chain (MHC) isoforms gene expression transition, blood pressure related parameters, calcium/calmodulin-dependent protein kinaseIIδ (CaMKIIδ), and monoamine oxidase (MAO) activities in rats' hearts. It was also planned to evaluate the effect of strenuous exercise on cardiac abnormalities induced by nandrolone. Thirty-two male wistar rats were assigned into four groups, namely control, nandrolone, nandrolone with strenuous exercise, and strenuous exercise groups. Nandrolone consumption significantly increased systolic, diastolic, pulse and dicrotic pressure, mean arterial pressure, as well as the amplitude of first peak (H1). Moreover, exercise combined with nandrolone completely masked this effect. The mRNA expression of ß-MHC and the ratio of ß -MHC/α -MHC showed a significant increase in the nandrolone and nandrolone with strenuous exercise groups compared to those in the control group. The values of heart tissue calcium/calmoldulin-dependent protein kinase IIδ (CaMKIIδ), and monoamine oxidase (MAO) in the nandrolone, nandrolone with strenuous exercise and exercise groups were significantly higher than those values in the control group. These findings indicate that nandrolone-induced heart and hemodynamic abnormalities may in part be associated with MHC isoform changes and Ca2+ homeostasis changes mediated by increased CaMKIIδ and MAO activities and that these effects can be provoked via strenuous exercise.

A Novel Electroactive Agarose-Aniline Pentamer Platform as a Potential Candidate for Neural Tissue Engineering.

Neuronal disorder is an important health challenge due to inadequate natural regeneration, which has been responded by tissue engineering, particularly with conductive materials. A bifunctional electroactive scaffold having agarose biodegradable and aniline pentamer (AP) conductive parts was designed that exhibits appropriate cell attachment/compatibility, as detected by PC12 cell seeding. The developed carboxyl-capped aniline-pentamer improved agarose cell adhesion potential, also the conductivity of scaffold was in the order 10-5 S/cm reported for cell membrane. Electrochemical impedance spectroscopy was applied to plot the Nyquist graph and subsequent construction of the equivalent circuit model based on the neural model, exhibiting an appropriate cell signaling and an acceptable consistency between the components of the scaffold model with neural cell model. The ionic conductivity was also measured; exhibiting an enhanced ionic conductivity, but lower activation energy upon a temperature rise. Swelling behavior of the sample was measured and compared with pristine agarose; so that aniline oligomer due to its hydrophobic nature decreased water uptake. Dexamethasone release from the developed electroactive scaffold was assessed through voltage-responsive method. Proper voltage-dependent drug release could be rationally expected because of controllable action and elimination of chemically responsive materials. Altogether, these characteristics recommended the agarose/AP biopolymer for neural tissue engineering.

Acceleration of skin wound healing by low-dose indirect ionizing radiation in male rats.

A recent hypothesis has revealed that low-dose irradiation (LDI) with ionizing radiation might have a promoting effect on fracture healing. The aim of this study was to investigate the influence of direct (electron beam) and indirect (gamma-ray) low-dose ionizing irradiations on the wound healing process in male rats. In 72 male rats, a full-thickness wound was incised. The animals were randomly assigned to three groups, each with 24 rats. The first two groups were named IG-I and IG-II and respectively exposed to electron and gamma-radiations (75 cGy) immediately after the surgical procedure. The third group was considered as the control (CG) and remained untreated. Skin biopsies from the subgroups were collected on days 3, 7, 15, and 21 after the operation and evaluated using histological and biomechanical methods. Data were analyzed by one-way ANOVA, followed by Tukey's post hoc test using SPSS 20 software. Histological studies of tissues showed that the mean number of fibroblasts, macrophages, blood vessel sections, and neutrophils on the third and seventh days after the surgery in the gamma-treated group was higher than that in both other groups. In contrast, on day 21, the mean number of mentioned cells in the gamma-treated group was lower than in the other two groups. In addition, the mean maximum stress value was significantly greater in the gamma-treated group. Results of this study showed that gamma-ray irradiation is effective in the acceleration of wound healing.

The Effect of Cerebrospinal Fluid in Collagen Guide Channel on Sciatic Nerve Regeneration in Rats.

AIM: The aim of this study was to evaluate the effect of cerebrospinal fluid (CSF) in nerve regeneration across the collagen guide channel in comparison with autograft. MATERIAL AND METHODS: Forty adult male rats (250-300 g) were randomized into (1) collagen channel+CSF, (2) collagen channel+normal saline (NS), (3) autograft, and (4) sham surgery groups. The left sciatic nerve was exposed and a 10 mm nerve segment was cut and removed. In the collagen groups, the proximal and distal cuts ends of sciatic nerve were telescoped into the nerve guides and CSF or NS injected into collagen conduit. In the autograft group, the 10 mm nerve segment was turned backwards and used an autologous nerve graft. All animals were evaluated by sciatic functional index (SFI) and electrophysiology, histology, and immunohistochemistry testing. RESULTS: The improvements in SFI since the beginning of the last evaluation in experimental groups were measured. On days 49 and 60 post-operation, the mean SFI of the collagen+CSF group was significantly greater than the autograft group (P < 0.05). On day 90, the mean nerve conduction velocity (NCV) of the collagen+CSF group was greater than autograft group (P < 0.05). The number of myelinated fibers in the collagen+CSF group was significantly greater than that of the collagen + NS group at day 90 (P < 0.05). CONCLUSION: CSF in collagen nerve guide channel effectively enhances nerve regeneration and promotes functional recovery in injured sciatic nerve of rats.

Evaluation of tendon healing using fibroblast like synoviocytes in rabbits: A biomechanical study.

Tendon never restores the complete biological and mechanical properties after healing. Several techniques are available for tissue-engineered biological augmentation for tendon healing like stem cells. Recently, synovium has been investigated as a source of cells for tissue engineering. In the present study, we investigated potentials of fibroblast like synoviocytes (FLSs) in tendon healing. Sixteen rabbits were divided randomly into control and treatment groups. One rabbit was used as a donor of synovial membrane (synovium). The injury model was unilateral complete transection through the middle one third of deep digital flexor tendon (DDFT). Subsequently, the tendon stumps were sutured with 3/0 nylon. In treatment group, 0.1 mL phosphate-buffered saline (PBS) solution containing 1 × 10(6) nucleated cells of FLSs was injected intratendinously at both tendon stumps just next to incision line. In control group, 0.1 mL PBS without FLSs was used for intratendinous injection. Model animals were euthanized at eight weeks, DDFTs were harvested and prepared for biomechanical study. Results of study showed that, there was no significant differences in biomechanical parameters values between FLSs treated and control groups. In conclusion, intratendinous injection of FLSs did not improve biomechanical properties during eight weeks in rabbit.

Alpha-lipoic acid loaded in chitosan conduit enhances sciatic nerve regeneration in rat.

OBJECTIVES: To investigate the effect of topical administration of alpha-lipoic acid into chitosan conduit on peripheral nerve regeneration using a rat sciatic nerve transection model. MATERIALS AND METHODS: Forty five Wistar rats were divided into three experimental groups randomly. A 10-mm gap of sciatic nerve was bridged with a chitosan conduit following surgical preparation and anesthesia. In treatment group, the conduit was filled with 30 µl alpha-lipoic acid (10 mg/kg/bw).It was filled with 30 µl phosphate buffered saline solution in control group. In Sham group sciatic nerve was just exposed. RESULTS: The recovery of nerve function was faster in treatment group than in control, at 4 and 8 weeks after surgery (P-value<0.05). Conduction velocity was better in treatment group than in control group at 4 and 12 weeks (P-value<0.05). Recovery index was higher in treatment group than the control group, 8 weeks after surgery (P-value <0.05). Greater nerve fiber diameter, axon diameter, and myelin sheath thickness were observed in treatment group compared to control group at 8 and 12 weeks after surgery (P-value<0.05). The immunoreactivity of regenerated axons and myelin sheath in treatment group were far more similar to sham group. CONCLUSION: Alpha-lipoic acid when loaded in a chitosan conduit could improve transected sciatic nerve regeneration in rat.

Premenstrual syndrome and quality of life in Iranian medical students.

PURPOSE OF STUDY: The purpose of this research was to investigate the prevalence of premenstrual syndrome (PMS) in medical students and to evaluate the hypothesis that PMS may result in a decrease in quality of life. METHODS: In a cross-sectional study, 142 female medical students who study at Urmia University of Medical Sciences were included. The data were compiled using a PMS questionnaire based on the fourth version (DSM-IV) criteria, the questionnaire of "Premenstrual Syndrome Scale" as well as the "World Health Organization's Quality of Life (WHOQOL-BREF)" questionnaire. FINDINGS: In total, 56 out of 142 (39.4%) female medical students met the DSM-IV criteria for PMS. In the PMS group, more than half of the girls, i.e. 60.6% had mild, 25.1% had moderate and 14.2% had severe PMS. PMS was found to be significantly high in students who have positive history of PMS in their first degree relatives and who have used drugs to relieve PMS symptoms (P<0.05). Life quality score was low in more than half of the medical students, especially in psychological and social components (P>0.05). However, the quality of life score means in mental health (P=0.02) and environmental health (P=0.002) decreases as the PMS score average increases. CONCLUSION: The results of premenstrual syndrome prevalence and their severity suggest that PMS is common in medical students and this adversely affects some domains of the quality of life. Improving the life quality of female medical students needs some interventions related to the PMS and also other interventions not related to PMS.

The mesenchymal stem cell-derived microvesicles enhance sciatic nerve regeneration in rat: a novel approach in peripheral nerve cell therapy.

BACKGROUND: The accomplishment for desired functional peripheral nerve regeneration is still challenging despite various materials and methods. The effects of local application of omental adipose mesenchymal stromal cell-derived microvesicles (MVs) on peripheral nerve regeneration were studied using a rat sciatic nerve transection model. METHODS: A 10-mm gap of sciatic nerve was bridged with a chitosan conduit. The rats were divided into five experimental groups randomly as follows: cultured undifferentiated omental adipose-derived stromal cells, rest mesenchymal stem cell-derived MVs (c-MVs), anti-inflammatory mesenchymal stem cell-derived MVs (anti-MVs), proinflammatory mesenchymal stem cell-derived MVs (pro-MVs), and negative control (Chit). RESULTS: The functional assessment of nerve regeneration (walking track analyses), electrophysiologic measurements, muscle mass measurements, as well as histomorphometrical and immunohistochemical indices showed drastic improvement in nerve regeneration in c-MVs and anti-MVs animals compared with pro-MVs animals (p < 0.05). CONCLUSION: The anti-inflammatory stem cell-derived MVs can be used as an alternative for the improvement of rat sciatic nerve regeneration.

Improvement of functional recovery of transected peripheral nerve by means of chitosan grafts filled with vitamin E, pyrroloquinoline quinone and their combination.

Effects of vitamin E and pyrroloquinoline quinone on peripheral nerve regeneration were studied using a rat sciatic nerve transection model. Ninety male healthy White Wistar rats were divided into three experimental groups (n = 15), randomly: Sham-operation (SHAM), transected control (TC), chitosan conduit (Chit) and three treatment groups (Vit E, PQQ and PQQ + Vit E). In SHAM group after anesthesia, left sciatic nerve was exposed through a gluteal muscle incision and after homeostasis muscle was sutured. In Chit group left sciatic nerve was exposed the same way and transected proximal to tibio-peroneal bifurcation leaving a 10-mm gap. Proximal and distal stumps were each inserted into a chitosan tube. In treatment groups the tube was implanted the same way and filled with Vit E, PQQ and PQQ + Vit E. Each group was subdivided into three subgroups of six animals each and were studied 4, 8, 12 weeks after surgery. Functional and electrophysiological studies, and gastrocnemius muscle mass measurement confirmed faster and better recovery of regenerated axons in Vit E + PQQ combination compared to Vit E or PQQ solely (P < 0.05). Morphometric indices of regenerated fibers showed number and diameter of the myelinated fibers in PQQ + Vit E was significantly higher than in other treatment groups. In immunohistochemistry, location of reactions to S-100 in PQQ + Vit E was clearly more positive than in other treatment groups. Response to PQQ + Vit E treatment demonstrates that it influences and improves functional recovery of peripheral nerve regeneration.

Using eggshell membrane as nerve guide channels in peripheral nerve regeneration.

OBJECTIVE(S): The aim of this study was to evaluate the final outcome of nerve regeneration across the eggsell membrane (ESM) tube conduit in comparison with autograft. MATERIALS AND METHODS: Thirty adult male rats (250-300 g) were randomized into (1) ESM conduit, (2) autograft, and (3) sham surgery groups. The eggs submerged in 5% acetic acid. The decalcifying membranes were cut into four pieces, rotated over the teflon mandrel and dried at 37(°)C. The left sciatic nerve was surgically cut. A 10-mm nerve segment was cut and removed. In the ESM group, the proximal and distal cut ends of the sciatic nerve were telescoped into the nerve guides. In the autograft group, the 10 mm nerve segment was reversed and used as an autologous nerve graft. All animals were evaluated by sciatic functional index (SFI) and electrophysiology testing. RESULTS: The improvement in SFI from the first to the last evalution in ESM and autograft groups were evaluated. On days 49 and 60 post-operation, the mean SFI of ESM group was significantly greater than the autograft group (P< 0.05). On day 90, the mean nerve conduction velocity (NCV) of ESM group was greater than autograft group, although the difference was not statistically significant (P> 0.05). CONCLUSION: These findings demonstrate that ESM effectively enhances nerve regeneration and promotes functional recovery in injured sciatic nerve of rat.

Desmopressin accelerates the rate of urinary morphine excretion and attenuates withdrawal symptoms in rats.

AIM: The aim of this study was to examine the effects of desmopressin on morphine withdrawal symptoms and vasopressin level in morphine-dependent subjects. METHODS: Wistar male rats were injected s.c. with morphine once per day for 5 consecutive days to induce morphine dependence. After morphine use ceased on day 5, an equal number of rats were assigned to one of four groups for either saline or desmopressin by either intraperitoneal (i.p.) or intracerebroventricular (i.c.v.) injection. From days 5 to 10, urine was collected daily and tested for the presence of morphine, and withdrawal symptoms were monitored to assess the effects of desmopressin. RESULTS: Significant weight loss occurred among all morphine-addicted rats during the withdrawal period. With both methods (i.p. and i.c.v.), the period of urinary morphine excretion was shorter for the two groups that were given desmopressin (experimental groups) than the two groups that were not given desmopressin (control groups), and no significant difference in urinary morphine excretion was found between the two experimental groups. During the early stage of withdrawal, the severity of the withdrawal symptoms in the experimental groups was significantly lower than that in the control groups. CONCLUSION: Desmopressin decreases the extent of morphine withdrawal symptoms, indicating that this agent might be appropriate for treating morphine addiction. Desmopressin appears to reduce withdrawal symptoms not by exerting an anti-diuretic effect but rather by exerting an effect on the central nervous system.

Long-term ethanol consumption initiates atherosclerosis in rat aorta through inflammatory stress and endothelial dysfunction.

Controversy exists on whether alcohol has a direct cardioprotective effect or it provokes atherosclerosis, so the present study sought to assess the effect of chronic consumption of ethanol on the markers of endothelial function, vessel rigidity, and atherosclerosis in the aorta of rat. Male Wistar rats were selected randomly and exposed to ethanol (4.5g/kg of 20% w/v solution in saline) once per day for 6weeks. Blood pressure, hemodynamic parameters, foam cell formation, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, endothelial-leukocyte adhesion molecule-1, and high-sensitivity C-reactive protein (CRP) were assessed in ethanol treated rats and compared with either sham or control rats. The results revealed a concurrent significant increase of adhesion molecules, CRP levels, systolic, diastolic, pulse, and dicrotic pressures as well as enhanced formation of foam cell in ethanol-treated rats. These findings implicate that long-term ethanol exposure provokes atherogenic and hemodynamic changes via significant induction of proinflammatory response, augmenting of cell adhesion molecules, stiffness in rat aorta wall and induction of foam cell formation.

Decreased blood pressure with a corresponding decrease in adhesive molecules in diabetic rats caused by vitamin E administration.

BACKGROUND: Hypertension is one of the important clinical problems of diabetic cardiovascular disease. The aim of this study was to determine the effect of vitamin E on blood pressure parameters and adhesive molecule amounts in diabetic rats. METHODS: Twenty-four male Wistar rats were divided into three groups (each of n = 8): the controls (C), non-treated diabetic (NTD), and vitamin E treated diabetic (VETD) groups. A single intraperitoneal injection of buffered streptozotocin (60 mg/kg) in cold sodium citrate (pH 4.5) was used to induce diabetes. The VETD group received 300 mg of vitamin E daily intragastrically for 6 weeks. Systolic and diastolic blood pressure, mean arterial pressure, as well as the dicrotic pressure, crest time, systolic and diastolic periods, and plasma levels of intercellular adhesion molecule-1 and E-selectin were measured after 6 weeks. RESULTS: The results revealed that there was a significant increase in systolic and diastolic blood pressures, mean arterial pressure, crest time, systolic duration, and the amount of sICAM-1 and E-selectin in diabetic rats. There was no significant difference in the heart rate or cardiac cyclic duration among the different groups. Significant improvement of blood pressure parameters as well as attenuation of the elevated ICAM-1 and E-selectin amounts was found in the vitamin E treated group. CONCLUSIONS: These findings indicate that vitamin E significantly improved blood pressure elevation in diabetic rats and that these effects could be associated with reducing adhesive molecule and antioxidant properties of vitamin E.

Reversible inactivation and excitation of nucleus raphe magnus can modulate tail blood flow of male Wistar rats in response to hypothermia.

BACKGROUND: The nucleus raphe magnus (NRM) is involved in thermoregulatory processing. There is a correlation between changes in the firing rates of the cells in the NRM and the application of the peripheral thermal stimulus. INTRODUCTION: we examined the effect of reversible inactivation and excitation of NRM on mechanisms involved in tail blood flow (TBF) regulation in hypothermia. METHODS: Hypothermia was induced in Male Wistar rats and cannula was implanted above the NRM. To evaluate the effect of nucleus inactivation on TBF, the amount of TBF was measured by Laser Doppler in hypothermic rats, before and after lidocaine microinjection into NRM. TBF was also measured after glutamate microinjection to assess the effect of nucleus excitation in hypothermic rats. RESULTS: Results indicated that after dropping TBF by hypothermia, microinjection of lidocaine into NRM significantly decreased TBF from 54.43 +- 5.7 to 46.81 +- 3.4, whereas glutamate microinjection caused a significant increase from 44.194 +- 0.6 to 98 +- 10.0 CONCLUSION: These data suggest that NRM have thermoregulatory effect in response to hypothermia.

Cardiovascular response to renin substrate microinjection into the central nucleus of the amygdala of rats.

Central nucleus of the amygdala is involved in cardiovascular regulation. Although most components of the renin-angiotensin system have been found to be distributed in amygdala, renin expression in brain has remained controversial. This work was undertaken to elucidate the extent of renin presence in this nucleus. A cannula was implanted bilaterally into the central nucleus of the amygdala. Mean arterial pressure and heart rate were directly measured via indwelling femoral artery cannula post bilateral intra central nucleus of the amygdala microinjection of renin substrate. Renin substrate microinjection dose-dependently increased mean arterial pressure and heart rate, whereas captopril, saralasin and losartan pretreatment inhibited these effects. The results suggest the presence of local renin or similar proteases in this nucleus.

Alteration of local ACE activity and vascular responsiveness during development of 2K1C renovascular hypertension.

OBJECTIVES: This study sought to examine the correlation between development of hypertension and local, including; aorta, heart, kidney, lung, as well as circulatory (serum) ACE activity in two kidney one clip (2K1C) renovascular hypertension. METHODS: Ten- to twelve-week-old rats undertaken left renal artery clipping. Experiments were carried out in 2, 4, 8, and 12 weeks after induction of hypertension (2, 4, 8 and 12W). After sacrificing, animals blood and tissues including heart, aorta lung and kidney were dissected out and homogenized in Trizma buffer. ACE activity was determined by hydrolysis of the synthetic substrate, Hip-His-Leu and the amount of hippuric acid liberated from the substrate were analyzed by HPLC. Vascular responsiveness was measured using perfusion pressure method. RESULTS: The systolic blood pressure (SBP) was gradually increased in 2K1C animals and was markedly higher compared to that of controls. The ACE activity in all tissues from 2K1C was significantly different in all groups of 2, 4, 8, and 12 weeks after surgery. The serum ACE activity of 2K1C was markedly increased in 2 and 4W and reached to plateau in 8 and 12W group. Vascular responsiveness to angiotensin I (AngI) was increased during development of hypertension in all groups of animals. CONCLUSION: These results indicated that there is a positive correlation between augmentation of blood pressure and local ACE activity in various tissues as well as serum, highlighting the significant contribution of local compared to circulatory ACE activity in development of renovascular hypertension.