Expression and Biological Evaluation of an Engineered Recombinant L-asparaginase Designed by In Silico Method Based on Sequence of the Enzyme from Escherichia coli.

Purpose: Medical usage of L-asparaginase (ASNase), the first-line of acute lymphoblastic leukemia treatment, is linked to allergic responses and toxicities, which necessitates the development of new bio-better ASNases. The aim of the current study was in silico design of a novel ASNase with predicted improved enzymatic properties using strategies encompassing sequence-function analysis of known ASNase mutants. Additionally, current study aimed to show that the new enzyme is active. Methods: Based on 21 experimentally reported mutations for ASNase, a virtual library of mutated enzymes with all 7546 possible combinations of up to 4 mutations was generated. Three-dimensional models of proposed mutant enzymes were built and their in silico stabilities were calculated. The most promising mutant was selected for preparing a genetic construct suitable for expression of the designed ASNase in bacterial cells. Results: Computational study predicted that Y176F/S241C double mutation of Escherichia coli ASNase may increase its folding stability. The designed ASNase was expressed in two different E. coli strains (Origami B(DE3) and BL21(DE3)pLysS) and then the soluble fractions prepared from the cell lysates of the host cells were used in enzyme activity assay. Results showed that enzyme activity of soluble fraction from Origami (95.4 ± 7.5 IU/0.1 mL) was four times higher than that of soluble fraction from pLysS (25.8 ± 2.5 IU/0.1 mL). Conclusion: A novel functional double mutant ASNase with predicted improved enzymatic properties was designed and produced in E. coli. The results of the current study suggest a great commercial potential for the identified enzyme in pharmaceutical and industrial applications.

Effects of Scrophularia oxysepala Methanolic Extract on Early Stages of Dimethylhydrazine-Induced Colon Carcinoma in Rats: Apoptosis Pathway Approach.

Purpose: Colorectal cancer is one of the most prevalent cancers, worldwide. The present study aimed to examine the effects of Scrophularia oxysepala (SO) methanolic extract on 1,2-dimethylhydrazine (DMH) induced colon cancer model in the Wistar rats. Methods: The animals administered DMH (40 mg/kg/S.C.) biweekly for 2 weeks to induce aberrant crypt foci (ACF). Other groups of animals were given the SO extract (50, 100 and 200 mg/kg/orally once/day) either before or after the DMH treatments. In the end, all animals were killed and at necropsy, the colon samples examined. The ACF, aberrant crypt (AC), crypt multiplicity (CM), caspase 3 protein and apoptosis measurement were performed. Results: The SO extract significantly (P<0.001) decreased the number of AC, ACF, and CM in all pre- and post-treated groups and caused significant increases in caspase 3 and apoptosis as compared to the DMH group. However, post-treated animals showed significantly more effective than pre-treatment groups. Methanolic extract of SO showed a chemopreventive potential, by effectively reducing the number of AC, ACF, and CM and increasing caspase 3 protein and apoptosis. Conclusion: One of the possible mechanisms might be involved in the induction of apoptosis through the caspase 3 mediated pathway.

Ultrasound elastography using shear wave interference patterns: a finite element study of affecting factors.

Elastography as one of the non-invasive medical imaging techniques which can help determine the stiffness of organs and other structures is currently attracting more attention. An interesting imaging rate-independent technique which has been discussed in literature uses shear wave interference patterns (SWIP). In this method, two external continuous harmonic vibration sources were used to induced SWIP and the resulting tissue displacements are mapped using ultrasonic imaging called sonoelastography. In this paper, a finite element model (FEM) of viscoelastic soft tissue with circular stiffer lesion inside, is simulated for testing the effect of stimulation characteristics on the propagation of SWIPs and shear speed map reconstruction. Also, we proposed an elastography probe, including miniature vibration sources and ultrasound transducer, which can be appropriate for experimental tests. The elastographic average speed ratio (ASR) and some scores like Dice coefficient, related to the binary image of shear speed map, are calculated for quantitatively measuring the effect of different contributing harmonic vibration parameters. Results show that the potential of providing useful diagnostic information can be improved if the preferable parameters are considered for implementation. According to these results the ASR, Dice and Jaccard scores would diverge from the ground truth of FEA if the parameter level is not selected correctly. Particularly, the Dice and Jaccard coefficients are obtained about 0.9 and 0.8, respectively, for the best vibration parameters level choice.

The Effects of Natural Clinoptilolite and Nano-Sized Clinoptilolite Supplementation on Lipid Profile, Food Intakes and Body Weight in Rats with Streptozotocin-Induced Diabetes.

Purpose: To determine the effect of natural clinoptilolite (CLN) and nano-sized clinoptilolite (NCLN) on lipid profile, food intakes (FI) and weight changes in streptozotocin (STZ) induced diabetic rats. Methods: In this experimental study, 36 rats were randomly divided into two groups: diabetic group which was injected STZ (60 mg/kg BW), and a non-diabetic group. Three days after diabetes induction, each of these groups was randomly divided into 3 subgroups of 6 animals ((1) control, (2) 1%/food CLN, (3) 1%/food NCLN). The animals were supplemented for 28 days, starting three days after STZ administration. At the end of the study, blood was drawn for biochemical assays. The weights and FIs of the rats were measured at the beginning and end of each week. Results: Our findings revealed that there was no significant change in lipid profile, 28 days after administration of STZ in diabetic rats. Low density lipoprotein (LDL) was increased slightly in diabetic rats treated with NCLN without any significant changes in other lipid profile parameters in the other groups. Weight was reduced significantly in diabetic rats. Administration of CLN and NCLN prevented further weight loss in diabetic rats. All groups treated with STZ had higher food intake during the study. Conclusion: Lack of beneficial changes in lipid profile may be attributed to short study duration, insufficient for appearance of lipid abnormalities. Given the partial improvement in weight status and lack of undesirable effects of clinoptilolite supplementation, further research is recommended in subjects with typ1 diabetes mellitus.

Semi-automatic measurement of rigid gas-permeable contact lens movement in keratoconus patients using blinking images.

PURPOSE: To introduce a method for estimation of the rigid gas-permeable contact lens (RGP) movement. MATERIALS AND METHODS: Videos captured from normal blinking of keratoconus patients while wearing RGP lenses were used for this study. The videos are recorded using the CCD camera of a smart phone attached to the eyepiece of the slit lamp. The algorithm starts with extracting two frames of the video related to the highest and lowest positions of the lens during blinking, followed by an appropriate edge detection method. In the next step circular Hough transform is used to find the center of lens and to segment it in each image. Finally the lens movement is estimated by measuring vertical displacement of the lens center between these two frames. RESULTS: Mean and standard deviation of the difference between real movement and results of the algorithm for 20 cases are -8.66% and 10.71% respectively. The results are highly correlated with Pearson coefficient 0.986 P < 0.001. Bland-Altman plot with 95% levels of agreement (LoA) shows an agreement between exact manual measurement method and the proposed algorithm. CONCLUSION: The proposed algorithm shows a relatively high accuracy as the first attempt and compared to the routine qualitative visual estimation. Considering the importance of the lens movement, although this system was not tested on a series of RGP fitting patients yet, semi-automatic measurement may potentially help practitioners decide the appropriate RGP lens fit and reduce the fitting time.

The Effects of Natural Clinoptilolite and Nano-Sized Clinoptilolite Supplementation on Glucose Levels and Oxidative Stress in Rats With Type 1 Diabetes.

OBJECTIVE: Oxidative stress has a major role in development of diabetic complications. In this study we investigated whether clinoptilolite and nano-sized clinoptilolite could reduce hyperglycemia and oxidative stress in streptozotocin-induced diabetic rats and attempted to determine which intervention was more effective. METHODS: Thirty-six rats were randomly allocated to 2 groups; 1 group was randomly chosen as a diabetic group and injected with streptozotocin (60 mg/kg body weight in 0.1 mol/L sodium citrate buffer, pH 4.5) to induce diabetes. Three days after diabetes induction, each group (diabetic group and nondiabetic group) was randomly divided into 3 subgroups of 6 animals each ([1] control, [2] 1% clinoptilolite/food, [3] 1% nano-sized clinoptilolite/food). Supplementation was continued for 28 days. Blood glucose was measured 3 times, at the beginning of the study and on the 14th and 28th days. Activity of antioxidant enzymes, including glutathione peroxidase and superoxide dismutase, and levels of total antioxidant capacity, as well as malondialdehyde, were evaluated. RESULTS: Blood glucose and malondialdehyde were significantly elevated, but there were no statistically significant changes in superoxide dismutase, glutathione peroxidase or total antioxidant capacity in diabetic rats. In diabetic rats treated with nano-sized clinoptilolite, blood glucose decreased to near normal levels (12.4 vs. 27.5 mmol/L). No significant changes were found in the other groups. None of the oxidative stress indices showed significant changes in either the treated or untreated rats. CONCLUSION: Nano-sized clinoptilolite exerted a hypoglycemic effect in streptozotocin-induced diabetic rats but had no significant influence on oxidative stress markers.

Apoptosis and Caspase 3 Pathway Role on Anti-Proliferative Effects of Scrophulariaoxy Sepala Methanolic Extract on Caco-2 Cells.

Colorectal cancer is one the most important malignancies worldwide and finding new treatment option for this cancer is of high priority. Natural compounds are common source of drugs for treatment of various diseases including cancers. The aim of this study was to investigate the effects of Scrophularia oxysepala extract on Caco-2 cells and explore the possible role of caspase 3 pathway in inducing cell death in this cancer cells in compare with chemotherapy agents of cisplatin and capecitabine. The methanolic extract of Scrophularia oxysepala (SO) was prepared by drench method. The IC50 of extract, cisplatin and capecitabine on Caco-2 cells were determined by MTT assay. The effect of SO extract on caspase 3 expression and inducing apoptosis were determined using TUNEL assay and caspase 3 ELISA methods, respectively. The IC50 of SO extract, cisplatin and capecitabine were 300, 195 and 80 µg/ml, respectively. Analysis for apoptosis revealed that SO methanolic extract increased apoptosis significantly (P<0.001) compared with control group. The effect of high doses of SO extract on apoptosis induction were comparable to cisplatin but significantly were higher than capecitabine. Only high doses of SO methanolic extract showed significant effects (P<0.05) on increasing caspase 3 compared to control group. The methanolic extract of SO showed inhibitory effect on Caco-2 cells and induced apoptosis in a dose-dependent manner comparable to cisplatin and higher than capecitabine 2 commonly used chemotherapeutic agent for various cancers.

Effects of a self-care education program on quality of life of patients with gastric cancer after gastrectomy.

BACKGROUND: Gastrectomy affects different aspects of functionality and impacts on the quality of life (QoL) of patients with gastric cancer. The importance of appropriate assessment of QoL in cancer patients is well established, yet strategies that help improve this important patient outcome are relatively scarce. OBJECTIVE: To examine the effectiveness of a brief self-care education program to improve QoL of gastric cancer patients after gastrectomy. METHODS: Using a randomized controlled trial, 59 patients with gastric cancer and candidate for gastrectomy were randomly assigned either to an intervention group (n = 31) to participate in a brief self-care education program or to a usual-care group (n = 28). Data were collected on patient demographics, and QoL was measured by the QLQ-C30 and the QLQ-STO22 at baseline and 1 month after gastrectomy. RESULTS: There were no statistically significant between-group differences in any subscales of the QLQ-C30 and the QLQ-STO22. However, participants in the brief self-care education program showed significant improvements from baseline in the global health status-QoL scale (t = 2.243, 𝘗 < .05), experience of pain (t = 2.508, 𝘗 < .05), constipation (t = 2.773, 𝘗 < .05), and the experience of dysphagia at the follow-up assessment. LIMITATIONS: This study is likely to be underpowered to show differences between the groups. CONCLUSION: A brief self-care education program was not sufficient to significantly improve the quality of life patients with gastric cancer after gastrectomy.

Effect of vitamin e on uroepithelial cells and changes of urinary sediments in oncology hospital nursing personnel.

INTRODUCTION: Vitamin E is an important natural antioxidant, and its most common and biologically active form is α-tocopherol. The antiproliferative effects of alpha-tocopherol have been previously demonstrated. In this study we investigated the effects of vitamin E on urinary epithelial cells and urinary sediments of nursing from oncology hospital. MATERIAL AND METHODS: Sixty-two female nursing personnel from oncology hospital participated in the study. They received orally 200mg of vitamin E per day for two weeks. Also prior to vitamin E and after vitamin E administration, the uroepithelial cells counts and other components of urinary sediments were carried out. RESULTS: There were significant differences in the epithelial cells count and treatment with vitamin E causing significantly more number of epithelial cells and urinary sediments to be excreted in the urine. DISCUSSION: Vitamin E significantly plays an important role on the excretion of uroepithelial cells and urinary sediments. CONCLUSION: In conclusion we propose that use of vitamin E at nontoxic levels would significantly enhance its antioxidative properties, especially among individuals subjected to prophylaxis of occupational hazards.

Anti-mutagenicity Effects of Vitamin E on Oncology and Non-oncology Hospital Nurses by Ames Assay.

INTRODUCTION: The aim of this study is to determine the anti-mutagenic effects of Vitamin E among nurses of oncology and non-oncology hospitals exposed to chemotherapy drugs. Several studies have demonstrated that nurses occupationally exposes to cytostatic drugs. MATERIAL AND METHODS: A total of 138 female nurses from oncology and non-oncology hospitals participated in the study. All urine samples of nurses before and after Vitamin E consumption (200 mg/day) were evaluated by Ames Salmonella typhimorium mutagenicity test using histidine negative of tester strain TA100 with and without S-9mix. In all steps the collected urine samples extracts were prepared using amberlit XAD-2 resins and examined for mutagenicity activity. The data of Ames assay were analyzed with Anova one way and t-test statistical. RESULTS: In the present study 25% of oncology nursing staff excrete carcinogenic compounds in their urine and oral consumption of Vitamin E for two weeks showed significant anti-mutagenic effects. DISCUSSION: It was appeared that the urinary mutagenic activity will decrease by receiving Vitamin E. However, after Vitamin E consumption there was significantly depletion of urinary mutagenic activity in urine extracts among the exposed nursing personnel. CONCLUSION: We conclude that mild effects of Vitamin E against poor safety and significant adverse events among nurses handling cytotoxic drugs. There is, therefore, a need to improve the safety of the work environment, make available protective equipment, develop standard practice guidelines for oncology nurses and higher therapeutic doses of Vitamin E may be a promising compound for reducing mutagenic effects of anti-neoplastic drugs among oncology hospital nurses.

Ambivalent effects of atorvastatin on angiogenesis, epidermal cell proliferation and tumorgenesis in animal models.

BACKGROUND: A growing body of preclinical data indicates that statins may possess antineoplastic properties; however, some studies have raised the possibility that statins may also have carcinogenic potential. METHODS: An air pouch model was used for angiogenesis. Single or multiple applications of croton oil on the back of Swiss albino mice with or without initiation by dimethylbenz(a)antheracene (DMBA) were used to evaluate the skin tumorgenesis, ultrastructural and histological alterations. RESULTS: Atorvastatin (orally, 10 mg/kg/day) produced a significant (P<0.05) reduction in angiogenesis. Concurrent administration of mevalonate reversed the anti-angiogenic effect of atorvastatin. However, local injection of atorvastatin (200 µg) into the pouches induced a significant (P<0.5) increase in angiogenesis that was not reversed by co-administration of mevalonate. The disturbance of cell polarity, inflammatory response, thickness of epidermal layer, and mitotic index induced by croton oil were inhibited markedly and dose-dependently (P<0.001) by pre-treatment with atorvastatin. In spite of the strong anti-inflammatory and anti-proliferative effects of atorvastatin on epidermal cell proliferation, it was identified that the same doses of atorvastatin in DMBA-initiated and croton oil-promoted skin tumorgenesis in mice increased the incidence of tumors and their conversion into malignant carcinoma. CONCLUSION: The reasons for these discrepancies remain unclear, and could be related to ambivalent effects of atorvastatin on angiogenesis or to specific differences in the experimental conditions. It is suggested that the pro-angiogenic effect of the drug, which could be responsible for promotion of skin tumors, is independent of the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibition that can be mediated directly by atorvastatin.

Effect of acute and chronic administration of carbamazepine on Cisplatin-induced hyperalgesia in rats.

BACKGROUND: Cisplatin is an effective antineoplastic drug used extensively in the treatment of malignancies. It induces painful peripheral neuropathy at high doses. OBJECTIVES: The aim of this study was to investigate the effect of carbamazepine (CBZ) on cisplatin-induced peripheral neuropathic pain by using the tail-flick test. MATERIALS AND METHODS: The study was performed using male Wistar rats weighing 180-200 g. Neuropathic pain was induced by intraperitoneal (IP) administration of cisplatin (5 mg/kg). The effect of oral (PO) CBZ administration (5, 10, and 15 mg/kg) on cisplatin-induced pain was assessed using the tail-flick test. RESULTS: Our results showed that cisplatin (5 mg/kg, IP) induced egregious pain (P < 0.01) on day 15. Acute administration of CBZ (5, 10, and 15 mg/kg, PO) caused significant (P < 0.05) increase in tail-flick time latency in a dose-dependent manner, in comparison with that observed in the control group. Furthermore, chronic administration of CBZ (5, 10, and 15 mg/kg, PO) increased (P < 0.05) the pain threshold on days 5 and 10. The analgesic effect of morphine (5 mg/kg, IP) was greater than that after acute CBZ administration (5, 10, and 15 mg/kg, PO). CONCLUSIONS: Our results showed that both acute and chronic CBZ administration attenuated cisplatin-induced pain. We suggest that CBZ can be used clinically for alleviating cisplatin-induced neuropathic pain in cancer patients, without any limitations such as tolerance to analgesic effect.

Buspirone attenuates tolerance to analgesic effect of morphine in mice with skin cancer.

Adjuvant drugs that can delay tolerance to morphine analgesia may lead to improved management of pain in chronic disease such as cancer. This study was aimed to investigate effect of buspirone, as a partial agonist of 5-HT1A receptor, on tolerance induced to morphine analgesic effect in animals with skin cancer. Study was carried on female Swiss albino mice. For skin tumorigensis, mice were treated with single dose of 7,12-dimethylbenz(a)anthracene (DMBA) and promoted by multiple dose of croton oil. Tolerance to morphine analgesia was induced by daily subcutaneous (sc) injection of morphine (5mg/kg for 30 days) and assayed by using the hot plate method. Results obtained from this study showed that pain threshold in mice with skin cancer were significantly lower. Tolerance to analgesic effect of morphine (5 mg/kg, sc) was appeared at day 15, whereas, in normal and skin tumor bearing mice co-treated daily with morphine (5 mg/kg, sc) and three different intraperitoneal (ip) doses of buspirone (5, 7.5 and 10 mg/kg) tolerance was observed at days 25 and 30. In conclusion our data indicate that concurrent use of morphine with buspirone may produce good cancer pain control and attenuate development of tolerance.

Effect of fluoxetine on tolerance to the analgesic effect of morphine in mice with skin cancer.

Adjuvant drugs that attenuate or inhibit the development of tolerance to morphine may lead to improved management of pain in chronic diseases such as cancer. The aim of this study was to investigate effect of fluoxetine, a specific 5-HT (5-hydroxytryptamine, serotonin) reuptake inhibitor, on tolerance induced to the analgesic effect of morphine in mice with skin cancer. The study was carried out on female Swiss albino mice. For skin tumorigensis, mice were initiated with a single dose of 7,12-dimethylbenz(a)anthracene (DMBA) and promoted by multiple doses of croton oil. Tolerance to morphine analgesia was induced by daily subcutaneous (sc) injections of morphine (5 mg/kg for 30 days) and assayed using the hot plate method. Results obtained from this study showed that pain thresholds in mice with skin cancer were significantly lower. Tolerance to the analgesic effect of morphine (5 mg/kg, sc) appeared at day 15, whereas in normal and skin tumor bearing mice co-treated daily with morphine (5 mg/kg, sc) and three different intraperitoneal (ip) doses of fluoxetine (0.16, 0.32 and 0.64 mg/kg) tolerance was observed at days 20, 25 and 30, respectively. In conclusion, our data indicate that concurrent use of morphine with fluoxetine may produce good cancer pain control and attenuate the development of tolerance.

Effect of testosterone on morphine withdrawal syndrome in rats.

AIM: To determine whether testosterone is involved in morphine withdrawal syndrome (WS). METHODS: In order to induce dependency, rats were treated with subcutaneous injection of morphine (days 1-2, 5 mg/kg; days 3-5, 7.5 mg/kg; days 6-8, 10 mg/kg), and after the last dose of morphine (day 8) WS was induced by intraperitoneal injection of naloxone (1 mg/kg). Wet dog shake (WDS), abdomen writhing (AW), and jumps (J) were recorded as indicators of WS. RESULTS: The severity of WDS, AW, and J in male rats was greater than that in females. Accordingly, in 4-week castrated and flutamide-treated (10 mg/kg/day for 8 days, i.p.) male rats, WDS, AW, and J were significantly decreased compared to male control rats. Testosterone replacement therapy (10 mg/kg/day for 8 days, i.m.) in 4-week castrated rats restored the severity of WDS, AW, and J behaviors to the level of non-castrated male rats, whereas testosterone potentiated the WDS behavior in non-castrated male rats. CONCLUSION: It can be concluded that testosterone might be effectively involved in morphine WS.

Assessment of anti-hyperlipidemic effect of Citrullus colocynthis

Hyperlipidemia is a well-known risk factor for several illnesses including atherosclerosis, heart and vascular diseases and stroke. In the search for potential anti-hyperlipidemic agents from plants to prevent these conditions, the pulp and the seeds of Citrullus colocynthis were assessed for their effects on the lipid profile of hyperlipidemic New Zealand rabbits. In the experimental groups that received the pulp of C. colocynthis or 100 mg/kg of seeds, the lipid profiles were significantly reduced when compared to the control group (P<0.05).

Hiperlipidemia é um fator de risco bem conhecido para diversas doenças inclusive aterosclerose, doenças cardíacas e derrame cerebral. Na procura de potenciais agentes antihiperlipidêmicos a partir de plantas para prevenir essas doenças, a polpa e as sementes de Citrullus colocynthis foram testadas para verificar seus efeitos no perfil lipídico de coelhos Nova Zelândia hiperlipidêmicos. Nos grupos experimentais que receberam a polpa de C. colocynthis ou 100 mg/kg das sementes, os perfis lipídicos foram significantemente reduzidos quando comparados ao grupo de controle (P<0,05).

Involvement of serotoninergic mechanism in analgesia by castration and flutamide, a testosterone antagonist, in the rat formalin test.

Several studies have suggested that testosterone has a role in nociception. Recently, we have shown that castration and flutamide, a testosterone antagonist, induce analgesia in the late phase of formalin test, which is related to increase of 5-HT levels in the dorsal horn of the lumbar spinal cord. The aim of the present study was to investigate the effect of fluoxetine, a selective serotonin reuptake inhibitor, on castration and flutamide-induced analgesia in order to further explore the role of 5-HT systems in such analgesia. Four weeks after castration, there was an analgesia in the late phase of formalin test, and this was potentiated by acute (0.32 mg kg(-1) ip) treatment of fluoxetine. Furthermore, coadministration of fluoxetine (0.32 mg kg(-1) ip) and flutamide (10 mg kg(-1) ip) produced more antinociceptive effect than those animals receiving fluoxetine and flutamide alone. The analgesic effect of fluoxetine (0.32 mg kg(-1) ip) and flutamide (10 mg kg(-1) ip) was abolished by pretreatment with 5,7-DHT (100 microg/rat it) and naloxone (2 mg kg(-1) ip). In summary, our data suggest that fluoxetine and flutamide have antinociceptive effects in tonic inflammatory pain through functional alteration of serotonergic systems, and their effects are potentiated by coadministration. The possible role of opioidergic system in their antinociceptive effect cannot be neglected.