Band 27, Heft 6

Background: Pain is the most common cause of patients seeking medical advice as a result of its association with different pathologies. This study evaluated the antinociceptive property of Haematostaphis barteri as well as the possible mechanism(s) associated with its antinociceptive property. Methods: Mice were administered H. barteri (30–300 mg kg −1 ; p.o.), followed by intraplantar injection of 10 μL of 5% formalin into the hind paws. The pain score was determined for 1 h in the formalin test. The possible nociceptive pathways involved in the antinociceptive action of H. barteri were determined by pre-treating mice with theophylline (5 mg kg −1 , a non-selective adenosine receptor antagonist), naloxone (2 mg kg −1 , a non-selective opioid receptor antagonist), glibenclamide (8 mg kg −1 ; an ATP-sensitive K + channel inhibitor), and atropine (3 mg kg −1 ; non-selective muscarinic antagonist). Results: H. barteri (30–300 mg kg −1 ) significantly and dose dependently precluded both first and second phases of nociception. Pre-treatment with naloxone had no effect on the analgesic activities of H. barteri in the first phase. Again, pre-treatment with atropine and glibenclamide did not significantly reverse the neurogenic antinociception of the extract in phase 1. However, theophylline reversed the analgesic effect of the extract in the first phase. In phase 2, theophylline had no effect on the analgesic activities of the extract. Naloxone, atropine, and glibenclamide significantly blocked the antinociception of H. barteri in the inflammatory phase of the formalin test. Conclusions: H. barteri possesses antinociceptive property mediated via the opioidergic, adrenergic, muscarinic, ATP-sensitive K + channels, and adenosinergic nociceptive pathways.

Background: Major depressive disorder (MDD) frequently co-occurs with other psychiatric problems. Our previous study showed that ATP1A1 gene expression level was significantly decreased in MDD patients. This research explores the potential correlations between the ATP1A1 expression level reduction and MDD patients’ clinical manifestation. Methods: All participant patients were diagnosed by Diagnostic and Statistical Manual of Mental Disorders – 4th edition (DSM-IV). Hamilton rating scale for depression (HAM-D) and anxiety (HAM-A) were applied to group patients into different categories. ATP1A1 expression level was measured by reverse transcript real-time polymerase chain reaction. Results: ATP1A1 expression levels of all MDD subgroups showed significant reduction compared to the control group (p<0.01). Further, the trend of ATP1A1 expression level reduction is significantly related to MDD patients’ HAM-A scores (p<0.01). However, there was no significance between ATP1A1 level and HAM-D scores (p>0.05). Conclusions: ATP1A1 expression level reduction is related to MDD anxiety score, which may be an explanation for the clinical manifestations and the underlining physiological mechanisms.

Background: The inflammatory response system has been implicated in the pathophysiology of major depression. The pro-inflammatory cytokines like interferon-γ induce the enzyme indoleamine-2,3-dioxygenase (IDO) of the kynurenine pathway of tryptophan metabolism. The induction of IDO reduces the availability of tryptophan for serotonin synthesis. Furthermore, the metabolites of kynurenine pathway have neurotoxic property, which along with decreased serotonin may account for depression-like illness. Methods: The aim of this study was to compare the effects of treatment with fluoxetine and 1-methyl-L-tryptophan (1-MT) on Bacillus Calmette-Guerin (BCG)-induced inflammatory model of depression in mice. Behavioral tests included locomotor activity, forced swim test (FST) and tail suspension test (TST). Oxidative stress was assessed by examining the levels of thiobarbituric acid reactive species (TBARS) and non-protein thiols (NP-SH) in homogenized whole brain samples. Comet assays were performed to assess neurotoxicity. Results: The results of this study demonstrate that BCG treatment resulted in an increase in duration of immobility in FST and TST as compared to the saline group. Further, it produced a significant increase in the brain TBARS levels and decrease in the brain NP-SH levels. The hippocampal tissue from BCG group had significantly more comet cells than the saline group. 1-MT and fluoxetine were able to reverse the BCG-induced depression-like behavior and the derangement in oxidative stress parameters. Fluoxetine and 1-MT also reversed the BCG-induced neurotoxicity in such mice. Conclusions: 1-Methyl-L-tryptophan exhibits antidepressant-like effect comparable to that of fluoxetine in treating BCG-induced depression-like behavior in mice.

Background: Smoking is a serious public health problem and an important risk factor of many diseases. The present study evaluated whether or not the influence of prolonged tobacco smoke (TS) exposure on spermatic indices and reproductive hormones would be reversible in young and adult rats. Methods: Prepubertal and adult rats were grouped into five (I, II, III, IV, and V) separately (n=5/group) and exposed to TS at target concentrations of 0, 0.5, 1.0, 0.5, and 1.0 mg nicotine/day, respectively for 30 days using the whole body exposure inhalation method. Groups I, II, and III were sacrificed 24 h after TS exposure, while groups IV and V were allowed to recover for 30 days before they were sacrificed. Results: Sperm count and motility were significantly (p<0.05) reduced in exposed prepubertal and adult rats. Additionally, sperm morphology was unaltered, testosterone was reduced, while luteinizing hormone (LH) and follicle stimulating hormone (FSH) were elevated compared to the non-TS exposed control group. The reductions in sperm count and motility were reversed only in adult recovery rats. LH and FSH elevations were reversed in all recovery animals, but testosterone concentrations remained lower than control. Furthermore, malondialdehyde levels in testes of exposed rats were significantly increased. This was reversed only in adult recovery rats that received 0.5 mg nicotine. Testicular levels of catalase, reduced glutathione, and superoxide dismutase were unaltered, except in prepubertal rats wherein catalase was decreased in both treated and recovery groups. Conclusions: The TS exposure alters sperm characteristics reversibly in adult, but irreversibly in prepubertal rats, which is attributable to elevation of oxidative stress.

Background: Smoking during pregnancy is associated with numerous fetal and developmental complications and reproductive dysfunctions in the offspring. Nicotine is one of the key chemicals of tobacco responsible for addiction. The present study was aimed to investigate the protective role of α-lipoic acid (ALA) during the transplacental nicotine-induced germ cell and DNA damage in the offspring of Swiss mice. Methods: Pregnant mice were treated with nicotine (20 mg/kg/day) in drinking water from 10 to 20 days of gestation period, and ALA (120 mg/kg/day) was administered orally for the same period. Endpoint of evaluation includes general observations at delivery and throughout the study, litter weight and size, sperm count and sperm head morphology, while structural damages and protein expression were assessed by histology and immunohistochemistry, respectively. Results: Maternal nicotine exposure led to decreased growth rate, litter and testicular weight, testosterone level, 3β-HSD expression and sperm count as well as increased sperm head abnormalities, micronucleus frequency and 8-oxo-dG positive cells, and the effects have been restored by ALA supplementation. Conclusions: The present study clearly demonstrated that ALA ameliorates nicotine-associated oxidative stress, DNA damage and testicular toxicity in the offspring by improving steroidogenesis, spermatogenesis and sperm count.

Background: Evidence shows that oxidative stress induced by nicotine plays an important role in bone loss. Vitamin E with its antioxidative properties may be able to reverse the effects of nicotine on bone. This study aimed to assess the effects of nicotine in the presence and absence of vitamin E on morphology, viability and osteogenic gene expression in MG-63 (osteosarcoma) human osteoblast-like cells. Methods: We treated the cells with 5 mM nicotine. The viability and morphology of cells were evaluated respectively using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) and crystal violet assays. The effect of nicotine on osteogenic gene expression in MG-63 cells was assessed by real-time reverse-transcription polymerase chain reaction of osteoblast markers, namely, alkaline phosphatase, osteocalcin and bone sialoprotein. Results: The results revealed that survival and proliferation of MG-63 cells were suppressed following exposure to nicotine, and cytoplasm vacuolization occurred in the cells. Nicotine significantly down-regulated the expression of osteogenic marker genes. Such adverse effects on morphology, viability and osteogenic gene expression of MG-63 cells were reversed by vitamin E therapy. Conclusions: In conclusion, vitamin E supplementation may play a role in proliferation and differentiation of osteoblasts, and vitamin E can be considered as an anabolic agent to treat nicotine-induced bone loss.

Background: Complex investigations of cytochrome P450 (CYP) isoforms with metabolic syndrome (MS) development are limited, and specific features of adolescent’s metabolisms are generally disregarded. The aim of present study was a comparative estimation of MS-mediated changes in CYP3A, CYP2C, and CYP2E1 mRNA expression and enzymatic activities, as well as antioxidant system parameters of adult and pubertal rats. Methods: Wistar albino male rats of two age categories [young animals of 21 days age (50–70 g) and adults (160–180 g)] were divided into four groups (eight animals in each group): (1) control 1 (intact young rats), (2) control 2 (intact adult rats), (3) MS3 (young rats with MS), and (4) MS4 (adult rats with MS). The MS was induced by full replacement of drinking water by 20% fructose solution (200 g/L). After 60 days of MS modeling, the investigation of rat liver CYP3A, CYP2C, and CYP2E1 mRNA expressions, their enzyme-marker activities, as well as the antioxidant system parameters was conducted. Results: Levels of liver CYP2E1 mRNA expression increased with MS: 40% (adults) and 80% (pubertal rats). Pubertal rats had also increased CYP3A2 mRNA expression (30%) and decreased CYP2C mRNA expression (30%). Changes in CYP2E1 and CYP2C enzymatic activities were consistent with the changes of corresponding gene expressions at both age-groups with MS. Simultaneously, liver reduced glutathione contents, and glutathione transferase and reductase activities were decreased in pubertal animals. Conclusions: CYP isoform expression rates and glutathione system were greatly violated with MS. The greater changes were observed in pubertal rats with MS.

Background: The purpose of this research was to study the effects of exercise on the concentration of plasma orexin A, a peptide regulating several physiological functions. Methods: Blood samples were collected from participants (men, n=10; age: 24.4±2.93 years) 15, 0 min before the start of exercise, and 30, 45, 60 min after a cycle ergometer exercise at 75 W for 15 min. Also heart rate (HR), galvanic skin response (GSR), and rectal temperature were monitored. Results: The exercise induced a significant increase (p<0.01) in plasmatic orexin A with a peak at 30 min after the exercise bout, in association with an increase of the other three monitored variables: HR (p<0.01), GSR (p<0.05), and rectal temperature (p<0.01). Conclusions: Our findings indicate that plasmatic orexin A is involved in the reaction to physical activity.

Background: Diabetic neuropathy is the most common complication of diabetes mellitus, and the different drug combinations available do not provide effective pain relief. The present study was performed to observe the effect of amitripyline, duloxetine, sitagliptin, and pregabalin, and their combinations on streptozotocin (STZ)-induced diabetic neuropathy. Methods: Diabetic neuropathy was induced by STZ, and the tail-flick test was used to assess thermal hyperalgesia before and after (at 30, 60, and 120 min) drug administration. One week after STZ administration, the blood glucose level was observed to be in the diabetic range. Results: Administration of all the drugs except sitagliptin increased the tail-flick latency significantly as compared to control. Further, the drugs amitriptyline, duloxetine, and pregabalin showed significant pain-relieving effect, when either two of them were administered in combination, although the different combinations had varied degree of pain relief. However, sitagliptin was observed to have no effect when administered alone or in combination with the other three drugs. Conclusions: Therefore, the study provides new insights concerning combined therapy of pain, which further needs clinical exploration.

Background: The duration of time for which the serum levels exceed the minimum inhibitory concentration (MIC) is an important pharmacokinetics (PK)/pharmacodynamics (PD) parameter correlating with efficacy for the antibiotic, ceftriaxone (CTRX). However, no reports exist regarding the PK or PD in patients undergoing continuous renal replacement therapy (CRRT). The purpose of this study was to examine the PK and safety of CTRX in patients undergoing CRRT in order to establish safer and more effective regimens. Methods: CTRX (1 g once a day) was intravenously administered four or more times to nine patients undergoing CRRT. Blood was collected after administration to measure CTRX concentrations in serum and the filtration fraction of CRRT by high-performance liquid chromatography. In addition to calculating PK parameters from serum CTRX, we (a) estimated by simulation CTRX concentrations when the dose interval was extended to once every 2 or 3 days, (b) calculated CTRX clearance via CRRT from CTRX concentrations in the filtration fraction, and (c) assessed the safety of CTRX use. Results: Total body clearance and the half-life of CTRX were 7.46 mL/min (mean) and 26.5 h, respectively, in patients undergoing CRRT. CTRX was found in the filtration fraction, and the estimated clearance by CRRT was about 70% of total body clearance. Simulations revealed that even when the dose interval is increased to 2 or 3 days, CTRX would retain its efficacy. Conclusions: Our findings suggest that, depending on the condition of patients undergoing CRRT, CTRX could be used safely against pathogens with a CTRX MIC ≤2 µg/mL, even when extending the dose interval.

Background: Erythrocyte swelling in non-ionic sucrose media and the subsequent osmotic lysis are influenced by mechanisms of regulatory volume adjustment and osmotic water permeability. Kinetics of transmembrane water and ion fluxes in varied physiologic states may determine the phenotype of erythrocyte osmotic fragility (EOF) and affect estimates of EOF. Methods: Effects of sex, age, late pregnancy (third trimester) and lactation on the haemolysis of Sahel goat erythrocytes incubated in a series of hyposmotic non-ionic sucrose media were investigated. Results: The fragiligram was sigmoidal in 72 (97%) out of 74 goats. Two male (3%) goats with low and high extreme median erythrocyte fragilities (MEF), had non-sigmoidal curves. The mean fragilities at osmolarities of 30–300 mosmol/L of sucrose and the mean osmolarities responsible for 10%–90% haemolysis (CH 10 –CH 90 ) were not significantly different between males and non-pregnant dry (NPD) females, amongst the age groups and between pregnant or lactating and NPD female goats. The MEF (CH 50 ) of the goats were at osmolarities of 126–252 mosmol/L (median of data: 171 mosmol/L) with a mean of 175.24±16.20 mosmol/L. Therefore, phenotypic homogeneity of EOF occurred with minor deviance, since EOF variables were not differentiated by sex, age, late pregnancy or lactation. Conclusions: Physiologic states of the goat did not affect EOF phenotype in non-ionic sucrose media. Sigmoidal fragility phenotype seemed to be homogeneously conserved by osmoregulatory mechanisms not partitioned by sex, age, late pregnancy or lactation, but a minor non-sigmoidal curve might have occurred due to altered erythrocyte osmotic behaviour that would require further investigation.

Background: Chronic arsenic exposure via contaminated drinking water is a global environmental health problem associated with hematological, hepatic and many serious systemic disorders. This study on adult male rats evaluated the protective effects of vitamin E (VE) and vitamin C (VC) against arsenic-mediated hematological and hepatic toxicities. Methods: Arsenic was administered orally as arsenic trioxide (3 mg/kg body weight/day), as a single dose for 30 consecutive days or along with VC/ascorbic acid (200 mg/kg body weight/day dissolved in water) and VE/α-tocopherol (400 mg/kg body weight/day dissolved in olive oil) as supplements. Multiple hematological and hepatic parameters were assessed. Results: Arsenic exposure caused significant reduction of erythrocyte counts (p<0.05), leukocyte counts (p<0.01) and hemoglobin (Hb) levels (p<0.01). Arsenic exposure also led to marked echinocytic transformation of erythrocytes resulting in increased morphological index (p<0.001). Altered serum oxidative balance was observed with a higher oxidative stress index (p<0.001). The results also showed a significant increase of serum cholesterol (p<0.05), low-density lipoprotein (p<0.001) and triglycerides (p<0.01), and decreased high-density lipoprotein (p<0.01) along with total protein (p<0.01). A marked elevation of hepatic thiobarbituric acid reactive substance (p<0.05) along with decreased reduced glutathione (p<0.001) levels were also observed. Interestingly, co-administration of VC and VE significantly prevented all the arsenic-induced alterations (p<0.05) except Hb content and serum protein. Conclusions: The present investigation offers strong evidence regarding the protective efficacy of co-administration of VC and VE against hematotoxicity and hepatotoxicity in adult male rats caused by chronic arsenic exposure.

Background: One of the most important problems in finding blood donors is the inadequacy of volunteer number. To overcome this problem, one of the solutions we suggest is innovating new health benefits of blood donation. The aim of the present study is to investigate the effects of blood donation on oxidative status markers and acute-phase reactants. Methods: A total of 96 healthy volunteers were recruited into the study. Blood samples were withdrawn 5 min before and 24 h after the blood donation. Serum nitric oxide, malondialdehyde levels, and activity of superoxide dismutase and myeloperoxidase were measured spectrophotometrically. Serum levels of high-sensitive C-reactive protein and pentraxin-3 as acute-phase reactants were measured by enzyme-linked immunosorbent assay kits. Results: We found statistically significant lower pentraxin-3 and high-sensitive C-reactive protein levels and higher superoxide dismutase activity and nitric oxide level 24 h after blood donation in serum of blood donor when compared with before blood donation. Conclusions: These findings suggest that blood donation affected oxidative status and acute-phase reactants in donors. Blood donation removes oxidants and decreases oxidative stress by elevating antioxidant enzyme such as superoxide dismutase. This is one more health benefit or reason why we should donate blood. Further large-scale studies should evaluate this mechanism and compare the same effect of wet cupping therapy.

Background: This study aims to investigate whether tested organic extracts possess antithrombotic properties with minimal or no toxicity and to predict the activity of some of their isolated compounds. Methods: An in vitro thrombolytic model was used to check the clot lysis effect of four Bangladeshi herbal extracts viz., roots of Curculigo recurvata W.T. Aiton (Satipata), leaf of Amorphophallus bulbifer Roxb. (Olkachu), leaf of Phyllanthus sikkimensis Muell. Arg., and whole plant of Thunbergia grandiflora Roxb. (Nillata) using streptokinase as a positive control and water as a negative control. Cytotoxicity was screened by brine shrimp lethality bioassay using vincristine sulfate as positive control. In silico prediction of activity spectra for substances (PASS) prediction was applied for phytoconstituents, namely, nyasicoside, glucomannan, grandifloric acid, serine, and alanine. Results: Using an in vitro thrombolytic model, C. recurvata , A. bulbifer , P. sikkimensis , and T. grandiflora showed 28.10±1.64%, 42.47±1.96%, 32.86±1.92%, and 25.51±1.67% of clot lysis, respectively. Reference drug streptokinase exhibited 75.00±3.04% clot lysis. Examined herbs showed significant (p<0.001) percentage (%) of clot lysis compared to negative control. In brine shrimp cytotoxic assay, C. recurvata , A. bulbifer , P. sikkimensis , and T. grandiflora showed LC 50 values 210.64±3.44, 98.51±1.47, 187.29±2.01, and 386.43±3.02 μg/mL, respectively, with reference to vincristine sulfate (LC 50 0.76±0.04). PASS predicted that examined phytoconstituents have a wide range of biological activity. Conclusions: Through our study it was found that A. bulbifer and P. sikkimensis could be considered as very promising and beneficial thrombolytic agents.