Different combinations of monoclonal antibodies and polyclonal antibodies in the design of neonatal hypothyroidism diagnostic kit.

Neonatal hypothyroidism is a deficiency of thyroid hormones at birth that can cause lifelong mental and physical disorders in humans. Lack of timely detection could lead to irreversible damage by neonatal hypothyroidism. However, it could be managed quickly and efficiently via timely diagnosis. The screening programs rely on immunoassays to diagnose neonatal hypothyroidism in most countries. This method is time-consuming, needs laboratory equipment, and should be performed by trained and skilled technicians. Given these circumstances, the ELISA method is not a preferable method for the diagnosing of neonatal hypothyroidism. However, it can be used as a confirmatory method in infants with suspected and unknown neonatal hypothyroidism. In the present study, the homemade SR95-1, SR95-2, and SR95-3 anti-ß-TSH polyclonal and the commercially available monoclonal antibodies were used to detect ß-TSH in a rapid assay kit design hypothyroidism screening. To design the kit, the different combinations of the antibodies were used to establish a sandwich immune-chromatography method. The designed rapid neonatal hypothyroidism tests were used to measure neonatal ß-TSH in 100 dry blood samples. This study showed that the best antibody pair in terms of sensitivity is the SR95-1 antibody as capture antibody and the SR95-2 as a conjugated antibody. Using 100 clinical samples, the designed assay was shown to have 94% sensitivity, 83% specificity, and 94% accuracy. The results showed that polyclonal antibodies (SR95-1 as capture) and SR95-2 (as detector) antibodies can detect the reference range of ß-TSH in dried blood samples and can be used in the screening of neonatal hypothyroidism.

A Metabolomic Investigation of Eugenol on Colorectal Cancer Cell Line HT-29 by Modifying the Expression of APC, p53, and KRAS Genes.

Colorectal cancer is one of the most lethal cancers with a high mortality rate. Chemotherapy results in drug resistance in some cases; hence, herbal medicines are sometimes used in adjunct with it. Eugenol has been reported to have anti-inflammatory, antioxidant, and anticancer properties. Metabolomics is a study of metabolic changes within an organism using high-throughput technology. The purpose of this research was to investigate the anticancer effects of eugenol and variations in p53, KRAS, and APC gene expression and metabolic changes associated with the abovementioned gene expressions using 1HNMR spectroscopy. The MTT method was used to determine cell viability and its IC50 detected. After treating HT-29 cells with IC50 concentration of eugenol, RNA was extracted and cDNA was obtained from them and the expression of p53, KRAS, and APC genes was measured using the qRT-PCR technique. Metabolites were extracted using the chloroform-ethanol method, lyophilized, and sent for 1HNMR spectroscopy using the 1D-NOESY protocol. Chemometrics analysis such as PLS-DA was performed, and differentiated metabolites were identified using the Human Metabolome Database. Integrated metabolic analysis using the metabolites and gene expression was performed by the MetaboAnalyst website. The observed IC50 for eugenol was 500 µM, and the relative expression of APC and p53 genes in the treated cells increased compared to the control group, and the expression of KRAS oncogene gene decreased significantly. The crucial changes in convergent metabolic phenotype with genes were identified. The results indicate that eugenol exhibits its antitumor properties by targeting a specific biochemical pathway in the cell's metabolome profile due to changes in genes involved in colon cancer.

The effect of Melissa officinalis L. extract on learning and memory: Involvement of hippocampal expression of nitric oxide synthase and brain-derived neurotrophic factor in diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetes is a systemic disease, which can cause synaptic defects in the hippocampus. Hippocampus plays a crucial role in learning and memory. Melissa officinalis L. has been used as for memory enhancement in Persian Medicine. AIM OF THE STUDY: The aim of this study was to evaluate the impact of the hydroalcoholic extract of Melissa officinalis L. on learning and memory, considering its impact on nitric oxide synthase and brain-derived neurotrophic factor expression in the hippocampus of diabetic rats. MATERIALS AND METHODS: Melissa officinalis L. extract was obtained by maceration method. To evaluate phenolic and flavonoid compounds of the extract, the samples were analyzed by HPLC. The animals were randomly divided into 6 groups: vehicle-treated control, Melissa officinalis-treated control (50 mg/kg), vehicle-treated diabetic, and M. officinalis-treated diabetic (25, 50, or 100 mg/kg). Diabetes was induced by streptozotocin And Melissa officinalis L. was administered for 2 weeks once diabetes was induced. Passive avoidance and Y-maze tasks were performed for learning and memory assessment. At the end of learning and memory tasks, rats were sacrificed and their hippocampus removed, lysed, and homogenized. The RNA contents were purified and then used as the template for cDNA synthesis. Real-time PCR was used to evaluate nitric oxide synthase and brain-derived neurotrophic factor genes expression. RESULTS: Rutin was main flavonoid compound and rosmarinic acid was the main phenolic compound of the Melissa officinalis extract. Streptozotocin induced diabetes and impaired learning and memory in diabetic rats. Melissa officinalis treated-control group showed a higher alternation score in the Y-maze task and step-through latency in the passive avoidance task compared to the vehicle treated diabetic group. Melissa officinalis-treated rats showed a higher alternation score in the Y-maze task in all doses compared to the vehicle treated diabetic group (P < 0.05). In addition, in the passive avoidance task Melissa officinalis increased step-through latency (P < 0.05) but not initial latency, in all doses. Furthermore, in diabetic rats, the expression of brain-derived neurotrophic factor and nitric oxide synthase genes decreased. However, hippocampal brain-derived neurotrophic factor and nitric oxide synthase gene expression was increased in Melissa officinalis-treated rats compared to diabetic rats (P < 0.05). CONCLUSIONS: Melissa officinalis improved learning and memory in diabetic rats, which may have occurred by increasing brain-derived neurotrophic factor and nitric oxide synthase gene expression.

Ginger Extract Reduces Chronic Morphine-Induced Neuroinflammation and Glial Activation in Nucleus Accumbens of Rats.

BACKGROUND: Chronic usage of morphine elicits the production of inflammatory factors by glial cells and induces neuroinflammation. Ginger (Zingiber Officinale Roscoe) is a medicinal herb that has anti-inflammatory properties. It has been reported that ginger shows anti-addictive effects against chronic usage of morphine; however, its influence on morphine-induced neuroinflammation has not yet been clarified. METHODS: Morphine (12 mg/kg) was administrated intraperitoneally for 6 consecutive days. To evaluate the effect of ginger on morphine-induced neuroinflammation, ginger extract (100 mg/kg) was given orally 30 minutes before morphine. Glial fibrillary acidic protein (GFAP) and p38 mitogen-activated protein kinase (p38 MAPK) levels were assayed by immunoblotting in the rat nucleus accumbens (NAcc). FINDINGS: The injection of chronic morphine increased the levels of proteins involved in neuroinflammation (p38 MAPK and GFAP) in NAcc. Furthermore, the levels of p38 MAPK and GFAP significantly returned to the control levels by ginger extract. CONCLUSION: The results suggest that the ginger extract can reduce morphine-induced neuroinflammation in NAcc.

Peganum Harmala L. Extract Reduces Oxidative Stress and Improves Symptoms in 6-Hydroxydopamine-Induced Parkinson's Disease in Rats.

Parkinson's disease is one of the most common neurodegenerative disorders. There are many documents about the effects of oxidative stress in Parkinson's disease etiology. Angiotensin II activates NADPH dependent oxidases and causes superoxides formation. Peganum harmala L. extract, which has angiotensin converting enzyme (ACE) inhibitory effect, is considered to evaluate oxidative stress inhibition and Parkinson's disease improvement. Male rats weighting 200-250 g were divided into 5 groups: Control, Neurotoxin (injection of 6-hydroxydopamine into left hemisphere substantia nigra), Peganum harmala's seeds aqueous extract (10 mg/kg) and captopril (5 mg/kg). Peganum harmala and captopril were injected intraperitonealy -144, -120, -96, -72, -48, -24, -2, 4 and 24 h relative to 6-hydroxydopamine injection time. Muscle stiffness, apomorphine induced unilateral rotation, amount of brain's protein oxidation and lipid peroxidation, ACE activity and histology of substantia nigra were assayed in all groups. Peganum harmala improved Muscle stiffness and one-direction rotation behavior significantly. It also reduced brain's lipid and protein oxidation levels in neurotoxin-injected rats significantly. In Peganum harmala group compared to control group, brain's ACE activity was significantly inhibited. In histological study, Peganum harmala prevented degeneration of dopaminergic neurons, too. In conclusion, aqueous extract of Peganum harmala could prevent symptoms and reduced oxidative stress markers in rats with Parkinson's disease induced by 6-hydroxydopamine.

Functional Recovery of Carbon Nanotube/Nafion Nanocomposite in Rat Model of Spinal Cord Injury.

BACKGROUND: Acute spinal cord injury (ASCI) can lead to paraplegia or quadriplegia, the treatment of which has been a major problem. New therapeutic approaches in developing carbon nanotubes (CNT) functionalized with the Nafion nanocomposite, a sulfonated tetrafluoroethylene copolymer, have been shown to increase the length of selected neurites in vitro. OBJECTIVE: We hypothesized that the administration of the CNT/Nafion nanocomposite after experimental SCI will promote regeneration of axons into the lesion cavity and the functional recovery of the hind limbs in a rat model. METHODS: To evaluate this hypothesis through this experimental research paper, transection SCI was induced at the T9-T10 vertebral level in adult female rats. One week after transection, the epicenter of the lesion was injected with 25 lL of vehicle (saline), or 1 lg/mL, 10 lg/mL, or 100 lg/mL of CNT/Nafion nanocomposite. Behavioral analysis was carried out by assessing tail flick, chronic pain or mechanical allodynia, motor coordination, and the results of the rotarod test performed pre- and post-surgery, on days 3, 7, 14, 21 and 28, using the tail flick analysis, Noldus CatWalk gait analysis, open-field locomotor test, and Rotarod test. At 28 days post-injection, the rats were euthanized and spinal cord tissue was extracted. RESULTS: We found that post-SCI, administration of the CNT/Nafion nanocomposite resulted in decreased lesion volume, increased neurofilament-positive fibers and corticospinal tract fibers in the lesion, and no increase in reactive gliosis (P < 0.001). Additionally, post-SCI administration of CNT/Nafion nanocomposite induced a modest improvement in hind limb locomotor recovery without inducing hyperalgesia. CONCLUSION: These data suggest that the CNT/Nafion nanocomposite may be an effective material to promote axonal repair and regeneration after SCI.

Fabrication and characterization of spongy denuded amniotic membrane based scaffold for tissue engineering.

OBJECTIVE: As a biological tissue material, amniotic membrane (AM) has low immunogenicity and to date has been widely adopted in clinical practice. However, some features such as low biomechanical consistency and rapid biodegradation is limited the application of AM. Therefore, in this study, we fabricated a novel three-dimensional (3D) spongy scaffold made of the extracellular matrix (ECM) of denuded AM. Due to their unique characteristics which are similar to the skin, these scaffolds can be considered as an alternative option in skin tissue engineering. MATERIALS AND METHODS: In this experimental study, cellular components of human amniotic membrane (HAM) were removed with 0.03% (w/v) sodium dodecyl sulphate (SDS). Quantitative analysis was performed to determine levels of Glycosaminoglycans (GAGs), collagen, and deoxyribonucleic acid (DNA). To increase the low efficiency and purity of the ECM component, especially collagen and GAG, we applied an acid solubilization procedure hydrochloridric acid (HCl 0.1 M) with pepsin (1 mg/ml). In the present experiment 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) cross linker agent was used to improve the mechanical properties of 3D lyophilized AM scaffold. The spongy 3D AM scaffolds were specified, by scanning electron microscopy, hematoxylin and eosin (H&E) staining, a swelling test, and mechanical strength and in vitro biodegradation tests. Human fetal fibroblast culture systems were used to establish that the scaffolds were cytocompatible. RESULTS: Histological analysis of treated human AM showed impressive removal of cellular components. DNA content was diminished after treatment (39 ± 4.06 µg/ml vs. 341 ± 29.60 µg/ml). Differences were observed between cellular and denude AM in matrix collagen (478 ± 18.06 µg/mg vs. 361 ± 27.47 µg/mg).With the optimum concentration of 1 mM NHS/EDC ratio1:4, chemical cross-linker agent could significantly increase the mechanical property, and resistance to collagenase digestion. The results of 2, 4, 6-Trinitrobenzenesulfonic acid (TNBS) test showed that cross-linking efficiency of AM derived ECM scaffolds was about 65% ± 10.53. Scaffolds treated with NHS/EDC cross-linker agent by 100 µg/ml collagenase, lost 75% of their dry weight after 14 days. The average pore size of 3D spongy scaffold was 160 µm measured from scanning electron microscope (SEM) images that it is suitable for cell penetration, nutrients and gas change. In addition, the NHS/ EDC cross-linked AM scaffolds were able to support human fetal fibroblast cell proliferation in vitro. Extracts and contact prepared from the 3D spongy scaffold of AM showed a significant increase in the attachment and proliferation of the human fetal fibroblasts cells. CONCLUSION: The extra-cellular matrix of denuded AM-based scaffold displays the main properties required for substitute skin including natural in vitro biodegradation, similar physical and mechanical characterization, nontoxic biomaterial and no toxic effect on cell attachment and cell proliferation.

Efficacy of Trigonella foenum-graecum Seed Extract in Reducing Metabolic and Inflammatory Alterations Associated With Menopause.

BACKGROUND: Several experimental and clinical studies support beneficial effects of Trigonella foenum-graecum (fenugreek) in the management of metabolic diseases and inflammatory disorders. OBJECTIVES: The purpose of this study was to examine the effect of T. foenum-graecum seed extract in reducing the metabolic and inflammatory alternations associated with menopause. MATERIALS AND METHODS: In this experimental study, 49 rats were divided into seven groups: (I) sham-control, (II) ovariectomized-control, (III and IV) ovariectomized treated with 50 and 150 mg/kg of T. foenum-graecum seed ethanolic extract, (V and VI) ovariectomized treated with 50 and 150 mg/kg of T. foenum-graecum hexanic extract, (VII) ovariectomized-positive control treated with 10 µg/kg of estradiol. The extracts were injected intraperitoneally one day after ovariectomy and the treatments were lasted for 42 days. RESULTS: Fasting blood glucose and body weight gain increased significantly in the ovariectomized-control group compared with that in the sham animals (P < 0.05). Administration of estradiol and T. foenum-graecum (50 and 150 mg/dL of hexanic extract and 150 mg/kg of ethanolic extract) significantly diminished the increase in glucose and body weight (P < 0.05). The serum level of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the ovariectomized control group was significantly higher than those in the sham animals (P < 0.05). Both hexanic and ethanolic extracts as well as estradiol were able to decrease level of these cytokines in the serum of ovariectomized rats (P < 0.05). CONCLUSIONS: The results of the present study show that administration of T. foenum-graecum corrects metabolic and inflammatory alterations associated with ovariectomy and has a potential for the management of menopause.

The Effect of Acute Intra Locus Coeruleus (LC) Microinfusion of Bupropion on Formalin-Induced Pain Behavior in Rat.

INTRODUCTION: Inflammatory pain is a common sign of chronic diseases. Some brain regions such as locus coeruleus (LC) of the brainstem nor-epinephrine (NE) system have a key role in The mechanisms of the pain modulation and dependence. Bupropion synthesized as an antidepressant, but it is using for smoke cessation. It can change morphine withdrawal signs such as pain related behaviors. This study tested the acute effect of intra-LC microinfusion of bupropion on the formalin-induced pain behavior in rats. METHODS: Wistar male rats were divided into 6 groups (control-naïve, control-operated, shamoperated, and 3 treated groups with 10(-2), 10(-3), 10(-4) mol/µl intra-LC of bupropion). The injection guide cannulae were implanted into LC nuclei bilaterally by stereotaxic coordinated surgery under sterile condition. The sham group received normal saline as drug vehicle but control groups had no intra-LC injections. Formalin (50 µl, 2.5%) was injected subcutaneously in plantar region of the right hindpaw in all animals (30 min after drug administration in treated animals). Nociceptive signs were observed continuously and registered on-line each minute. Common pain scoring was used for pain assessment. RESULTS: The analysis of data by one-way ANOVA showed that bupropion can reduce pain behavior scores significantly. Bupropion reduced total pain score in the phase 01 (60%) and phase 02 (52%) of maximal behavior compared to the sham group, dose dependently and significantly. The pain scores of controls and sham groups had no significant difference. DISCUSSION: The results showed that bupropion has analgesic effects on LC neurons and can alter the neurochemical involvement of LC in pain process. Bupropion has different and significant effect on early and late phases of formalin test.

Ginger (zingiber officinale roscoe) prevents morphine-induced addictive behaviors in conditioned place preference test in rats.

BACKGROUND: Consumption of chronic morphine induces neuro-inflammation and addictive seeking behavior. Ginger (Zingiber Officinale Roscoe), a well-known spice plant, has been used traditionally in the treatment of a wide variety of ailments. It has been shown that ginger has anti-inflammatory, anti-oxidative and antinociceptive properties. However, its influences on morphine-induced addictive behaviors have not yet been clarified. The aim of the present study was the inhibition of exploratory behavior of morphine addiction in the conditioned place preference test in male desert rats through ginger. METHODS: For conditioning to the morphine, the male Wistar rats received morphine (12 mg/kg intraperitoneally or i.p.) for 6 consecutive days and treatment groups were given different doses of ginger (25, 50 and 100 mg/kg intragastrically or i.g.) 30 min before morphine injection. For investigating addictive seeking behavior, conditioned place preference test (CPP) was used. FINDINGS: Our result demonstrated that injection of morphine for 6 days induces dependency to morphine and creates addictive seeking behavior and ginger (100 mg/kg) could decrease time spend in conditioning box (addictive seeking behavior). CONCLUSION: The data indicated that ginger extract has a potential anti-addictive property against chronic usage of morphine.

Inhibitory effect of Thymus caramanicus Jalas on hyperglycemia-induced apoptosis in in vitro and in vivo models of diabetic neuropathic pain.

ETHNOPHARMACOLOGICAL RELEVANCE: Since Thymus caramanicus Jalas is used as a folk medicine for the treatment of rheumatism, skin disorders, bacterial infections and diabetes and it contain antioxidant agents, we decided to investigate the possible effects of Thymus caramanicus Jalas (TCJ) extract on in vitro and in vivo models of diabetic neuropathy. MATERIALS AND METHODS: The high glucose-induced cell injury in Pheochromocytoma (PC12) cells and streptozotocin-induced diabetic rats were used. Tail-flick and rotarod treadmill assessments were used to determine nociceptive threshold and motor coordination. Cell viability was determined by MTT assay test. Western blotting was performed to measurement of apoptosis markers. RESULTS: The data showed that elevation of glucose consecutively increases functional cell injury and apoptosis. Furthermore, diabetic rats developed thermal hyperalgesia and motor deficit. Activated caspase 3, cytochrome c release and Bax/Bcl-2 ratio were significantly increased in high glucose-treated PC12 cells and in spinal cord of diabetic animals. TCJ extract (60 and 80 µg/ml) attenuates high glucose-induced PC12 cells damage and apoptosis. In diabetic animals, TCJ extract at daily doses of 100 and 150 mg/kg ameliorated hyperalgesia and suppressed spinal apoptosis. CONCLUSION: The data indicate that TCJ extract has neuroprotective effects against high glucose-induced neural damage. These protective effects are mediated, at least in part, through attenuation of neural apoptosis and suggest therapeutic potential of TCJ extract in amelioration of diabetic neuropathy.

Effect of noise stress on count, progressive and non-progressive sperm motility, body and genital organ weights of adult male rats.

AIMS: It was decided to investigate the effect of noise pollution on the body weight, genital organ weights, and also on sperm parameters. SETTING AND DESIGN: It is a prospective study designed in vitro. MATERIALS AND METHODS: A total 20 adult male wistar rats were used in this study. All rats were divided into 2 equal groups (n = 10): (1) control group and (2) experimental group. Animals of the experimental group were exposed to noise for 50 days with an intensity of 90-120 db and frequency of 300 - 350 Hz for 12 hours daily. After 50 days, at first, body weights of all animals were recorded, and then they were killed. The right epididymides were removed and also, sperm concentration and motility were determined. Each organ was weighed separately on an electronic balance. STATISTICAL ANALYSIS USED: Data are reported as mean ± SD and percentage. The statistical significance of difference between the control and experimental groups was determined by the unpaired t-test. RESULTS: The weights of the testes, epididymes, seminal vesicle, ventral prostate were found to be significantly decreased in rats exposed to noise pollution when compared with the weights of the same organs obtained from control group (P < 0.05). There was a statistical difference of P < 0.05 between the 2 groups in terms of sperm concentration. CONCLUSIONS: It is concluded that noise pollution has the bad effects on sperm concentration and motility; therefore, it is supposed that homes and places of working must be build far away of noisy of factories and other places with noise.

Chronic cyanidin-3-glucoside administration improves short-term spatial recognition memory but not passive avoidance learning and memory in streptozotocin-diabetic rats.

This research study was conducted to evaluate the efficacy of chronic cyanidin-3-glucoside (C3G) on alleviation of learning and memory deficits in diabetic rats as a result of the observed antidiabetic and antioxidant activity of C3G. Male Wistar rats were divided into control, diabetic, C3G-treated-control and -diabetic groups. The C3G was administered i.p. at a dose of 10 mg/kg on alternate days for eight weeks. For evaluation of learning and memory, initial latency (IL) and step-through latency (STL) were determined at the end of study using passive avoidance test. Meanwhile, spatial recognition memory was assessed as alternation in the Y-maze task. Oxidative stress markers in brain tissue were also measured. It was found that the alternation score of the diabetic rats was lower than that of control (p < 0.01) and C3G-treated diabetic rats showed a higher alternation score as compared to diabetic group (p < 0.05). Diabetic rats also developed a significant impairment in retention and recall in passive avoidance test (p < 0.01) and C3G treatment of diabetic rats did not produce any significant improvement. Meanwhile, increased level of malondialdehyde (MDA) in diabetic rats was significantly reduced following C3G treatment (p < 0.05). Taken together, chronic C3G could improve short-term spatial recognition memory disturbance in the Y-maze test but not retention and recall capability in passive avoidance test in STZ-diabetic rats.

Protective effect of Nigella sativa and thymoquinone on serum/glucose deprivation-induced DNA damage in PC12 cells.

OBJECTIVE: The discovery and development of natural products with potent antioxidant properties has been one of the most interesting and promising approaches in the search for treatment of CNS injuries. The most significant consequence of the oxidative stress is thought to be the DNA modifications, which can become permanent via the formation of mutations and other types of genomic instability resulting cellular dysfunction. Serum/glucose deprivation (SGD) has served as an excellent in vitro model for the understanding of the molecular mechanisms of neuronal damage during ischemia and for the development of neuroprotective drugs against ischemia-induced brain injury. Nigella sativa (N. sativa) seeds and thymoquinone (TQ), its most abundant constituent, have been shown to possess anti-inflammatory, antioxidant, chemopreventive and anti-neoplastic effects both in vitro and in vivo. Therefore, in this study we investigated genoprotective effects of N. sativa and TQ on DNA damage of PC12 cells under SGD condition. MATERIALS AND METHODS: PC12 cells were cultured in DMEM medium containing 10% (v/v) fetal bovine serum, 100 units/ml penicillin, and 100 µg/ml streptomycin. Initially cells were pretreated with different concentrations of N. sativa extract (NSE), (10, 50, 250 µg/ml) and TQ (1, 5, 10 µg/ml) for 6 h and then deprived of serum/glucose (SGD) for 18 h. The alkaline comet assay was used to evaluate the effect of these compounds on DNA damage following ischemic insult. The amount of DNA in the comet tail (% tail DNA) was measured as an indicator of DNA damage. RESULTS: A significant increase in the % tail DNA was seen in nuclei of cells following SGD induced DNA damage (p<0.001). In the control groups, no significant difference was found in the % tail DNA between NSE- or TQ-pretreated and vehicle-pretreated PC12 cells (p>0.05). NSE and TQ pretreatment resulted in a significant decrease in DNA damage following ischemic insult (p<0.001). This suppression of DNA damage by NSE and TQ was found to be dose-dependent. CONCLUSION: These data indicate that NSE and TQ have a genoprotective property, as revealed by the comet assay, under SGD condition in PC12 cells.

Vascular mechanisms of cyanidin-3-glucoside response in streptozotocin-diabetic rats.

BACKGROUND AND OBJECTIVE: Considering the high incidence of cardiovascular disorders in diabetes mellitus and some evidence on the antioxidant and antidiabetic potential of cyanidin-3-glucoside (C3G), this study was conducted to evaluate the possible beneficial effect of C3G administration on vascular reactivity of isolated thoracic aorta in diabetic rats and some of its underlying mechanisms. MATERIALS AND METHODS: Male diabetic rats received C3G (10mg/kg; i.p.) on alternate days for 8 weeks one week after streptozotocin (STZ) diabetes induction. RESULTS: It was found out that treatment of diabetic rats with C3G exerted a hypoglycaemic effect and attenuated the increased malondialdehyde (MDA) content and reduced the activity of superoxide dismutase (SOD) in aortic tissue. Maximum contractile response of endothelium-intact aortic rings to phenylephrine (PE) was significantly lower in C3G-treated diabetic rats relative to untreated diabetics and endothelium removal abolished this difference. Meanwhile, endothelium-dependent relaxation to acetylcholine (ACh) was significantly higher in C3G-treated diabetic rats as compared to diabetic group. CONCLUSION: Chronic treatment with C3G may prevent some diabetes-related changes in vascular reactivity observed in diabetic rats directly and/or indirectly due to its hypoglycaemic effect and attenuation of lipid peroxidation and through endothelial-derived factors.

The effects of dopaminergic drugs in the dorsal hippocampus of mice in the nicotine-induced anxiogenic-like response.

RATIONALE: Nicotine, an active alkaloid of tobacco has an acetylcholine property that alters anxiety-like behaviors in rodents. Moreover, several investigations suggest that the mesolimbic/cortical dopamine systems to be involved in the drugs affecting anxiety. The dopaminergic modulation of acetylcholine synaptic transmission has also been also suggested by different studies. Furthermore, modulation of such behaviors in rodents may be mediated through the dorsal hippocampus. OBJECTIVES: In the present study, a possible role of the dorsal hippocampal acetylcholine receptor mechanism in nicotine's influence on anxiogenic-like responses has been investigated. METHODS: During test sessions, the hole-board was used to investigate the effects of SCH23390, sulpiride, SKF38393 and quipirole on nicotine response in mice. RESULTS: Intraperitoneal (i.p.) administration of nicotine (0.5 mg/kg) decreased the number of head dips but had no effect on other behaviors. Intra-dorsal hippocampal injections of ineffective doses of SCH23390 (SCH; 0.125 and 0.25 µg/mouse) or sulpiride (SUL; 0.5 and 0.75 µg/mouse) reversed head dips induced by nicotine but did not impact other exploratory behaviors. Furthermore, co-administration of ineffective doses of SKF38393 (SKF; 4 µg/mouse, dorsal hippocampus) or quipirole (QUI; 0.5 µg/mouse) in conjunction with an ineffective dose of nicotine (0.25 mg/kg, i.p.) decreased head dips induced by nicotine, but were otherwise ineffective. CONCLUSION: These results may indicate a modulatory effect for the dorsal hippocampus dopamine receptors (D1 and D2) on an anxiogenic-like response induced by nicotine.

Mercury absorption by Pseudomonas fluorescens BM07 grown at two different temperatures.

Pseudomonas fluorescens BM07 was characterized as a producer of cold-induced biopolymer by decreasing the temperature down to as low as 10 degrees C. It was previously shown that the synthesis of BM07 biopolymer was inhibited at 30 degrees C. The present study was conducted to investigate the biosorption of mercury (Hg2+) ions on the BM07 cells grown on M1 minimal medium at two temperatures (10 degrees C and 30 degrees C). The effects of various factors including pH, contact time, initial concentration of metal and cell biomass on the biosorption yield were also studied. Study of the effect of pH on mercury removal indicated that the metal biosorption increased with increasing pH from 3.0 to 7.0. The optimum adsorption pH value was found to be 7.0. Our results showed that, at optimum pH, BM07 cells were able to uptake the mercury up to 102 and 60 mg Hg2+/g dry biomass for 10 degrees C and 30 degrees C grown cells respectively. The removal capacity of cells increased when the cell biomass concentrations increased. The maximum removal efficiency was obtained when cells concentration was 0.83 mg dry biomass/ml for both conditions. The initial metal ion concentration significantly influenced the equilibrium metal uptake and adsorption yield. The equilibrium data were analyzed using Langmuir adsorption model. The qmax was 62.9 and 82.25 mg Hg2+/g dry biomass for cells grown at 30 degrees C and 10 degrees C respectively. The results suggest that, the existence of residual cold-induced biopolymer on the external surface of cells may play an important role in biosorption efficiency, as P. fluorescens BM07 cells which were grown at 10 degrees C under similar conditions showed higher efficiency to biosorbe mercury than non-polymer producing cells grown at 30 degrees C.

The effects of Vitex agnus castus extract and its interaction with dopaminergic system on LH and testosterone in male mice.

The purpose of this study was to evaluate the probable effects of Vitex agnus castus (Vac.) on the male reproductive physiology. It is a well known fact that LH secretion from the anterior pituitary of mammals is controlled by many neurotransmiters such as dopamine. In this experiment, we have studied the effect of Vac. extract on the LH and testosterone hormones and its interaction with the dopaminergic system on male mice. In order to evaluate these effects, we used the hydroalcoholic Vac. extract (for extraction we used percolation technique) injection with the following doses: 65, 165, 265, 365 and 465 mg kg-', bromocriptine as a dopamine receptor agonist (5, 10, 20 mg kg(-1)) and haloperidol as a dopamine receptor antagonist (1, 1.5, 2, 2.5, 3 mg kg(-1)). To study the interaction between Vac. extract and dopaminergic system, we injected the optimum doses of Vac. with bromocriptine or haloperidol at the same time. Intraperitoneal injections were applied in all experiments, once a day for 30 days. The control group remained intact and the sham group received vehicle. After the last injection, we collected the animal blood serums for hormonal assays. LH and testosterone were measured by Radio Immuno Assay (RIA). LH and testosterone, showed significant decrease in bromocriptine group and haloperidol increased these hormones. Vac. extract decreased significantly the LH and testosterone levels. The coadministration of Vac. extract and bromocriptine decreased LH and testosterone. Coadministration of Vac. extract and haloperidol decreased LH and testosterone levels. These results suggest: dopamine regulates the gonadotroph-leydig cells axis. It appears that Vac. exertes effects through dopaminergic system and other pathways. The findings of this study show we can use Vac. extract for pathological cases of increasing LH and testosterone.