Inotropic and chronotropic effects of 6-hydroxy-4-methylquinolin-2(1H)-one derivatives in isolated rat atria

Background: Selective phosphodiesterase (PDE3) inhibitors improve cardiac contractility and may use in congestive heart failure. However, their proarrhythmic potential is the most important side effect. Methods: In this research, we evaluated the potential cardiotonic activity of six new synthesized selective PDE3 inhibitors (6-hydroxy-4-methylquinolin-2(1H)-one derivatives) using the spontaneously beating atria model. In each experiment, atrium of reserpine-treated rat was isolated and the contractile and chronotropic effects of a synthesized compound were assessed. The 3-isobutyl-1-methylxanthine, a non-selective PDE inhibitor, was used for comparison. Results: The results showed that, among new compounds, the best pharmacological profile was obtained with the compound 6-[4-(4-methylpiperazine-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one, C6, which displayed selectivity for increasing the force of contraction (168 +/- 5% change over the control) rather than the frequency rate (138 +/- 5% change over the control) at 300 muM. However, C6 at concentrations of 10 and 100 muM produced left and upward shift in the positive inotropic concentration-response curve of isoprenaline. The -log EC50 of isoprenaline was 8.843 +/- 0.171 in the absence, 9.448 +/- 0.138 and 9.456 +/- 0.107 in the presence of 10, 100 muM of C6, respectively (P<0.001, n = 6). Also, amrinone, a selective PDE3 inhibitor, shifted the isoprenaline concentration-response curve to the left and upward. The concentration of 10 and 100 muM amrinone decreased -log EC50 of isoprenaline to 9.527 +/- 0.287 and 9.423 +/- 0.243, respectively (P<0.001, n = 6). Moreover, the positive chronotropic effect of isoprenaline was not affected by amrinone or C6. Conclusion: This study provides functional evidence for the positive inotropic effect of C6. Considering the augmentation of isoprenaline positive inotropic concentration-response with C6 and amrinone, we conclude that C6 produces its effect via potentiation of cAMP-dependent signaling system and possibly by inhibition of PDE3 activity.     

Differentiation of human umbilical cord mesenchymal stem cell into germ-like cell under effect of co-culture with testicular cell tissue

Supplements produced by mouse testicular cells (mTCs) and the interaction between cells can increase the differentiation rate of human umbilical cord mesenchymal stem cells (hUCMSCs) into the germ-like cells. We studied the differentiation rate of hUCMSCs into the germ-like cells under effect of mTCs co-culturing. Isolated hUCMSCs from postpartum human umbilical cords were cultured. Then, the expression of mesenchymal (CD73, CD90 and CD105) and haematopoietic (CD34 and CD45) markers of hUCMSCs were confirmed by flow cytometry. Then, the hUCMSCs were cultured in four distinct groups: (a) control, (b) co-culture until D0, (c) co-culture until D5 and (d) co-culture until D10, in order to differentiate into the germ-like cells. After 10 days, the expression of OCT4, VASA, Fragilis and SYCP3 genes were examined by Real-Time qPCR. The flow cytometry indicated a high expression of mesenchymal markers and a low expression of haematopoietic markers (CD73:98.6%, CD90: 99.1%, CD105: 99.5%, CD34: 4.22% and CD45: 2.54%). The expression of OCT4 decreased during the time while the expression of VASA, Fragilis and SYCP3 markers increased in the co-culture with testicular cells (p value <.05). Co-culture with mTCs may be used as an effective method to differentiate hUCMSCs into germ-like cells.     

The response of wildtype and mutant cultivars of soybean to salt stress - comparing vegetative and reproductive phases on the basis of leaf biochemical contents,RWC, and stomatal conductance

Soybean, as a major oilseed crop, is cultivated in many parts of the world including saline-affected regions. A four-replicated factorial experiment based on a completely randomized design with five salinity levels (control, 50, 100, 150, and 200 mM NaCl) and five wild-type and new mutant cultivars of soybean (Williams, Clark, M-4, M-7, and M-9) was done at two vegetative and reproductive phases to study leaf oxidants, antioxidants and osmolytes under salinity stress. The vegetative phase demonstrated higher K⁺:Na⁺ ratio, cell membrane stability, relative water content (RWC) and stomatal conductance to H₂O (gs) accompanied with lower oxidative stress, antioxidants and osmolytes contrasting to reproductive phase. At both phenological phases, saline stress stimulated antioxidative defense owing to the enhancement of oxidative stress. The results indicated that mutants possessed a better antioxidative system than their wild-types.     

Transplanting P75-Suppressed Bone Marrow Stromal Cells Promotes Functional Behavior in a Rat Model of Spinal Cord Injury

Background: Bone marrow stromal cells (BMSC) have been successfully employed for movement deficit recovery in spinal cord injury (SCI) rat models. One of the unsettled problems in cell transplantation is the relative high proportion of cell death, specifically after neural differentiation. According to our previous studies, p75 receptor, known as the death receptor, is only expressed in BMSC in a time window of 6-12 hours following neural induction. Moreover, we have recently reported a decreased level of apoptosis in p75-suppressed BMSC in vitro. Therefore, our objective in this research was to explore the functional effects of transplanting p75:siRNA expressing BMSC in SCI rats. Methods: Laminectomy was performed at L1 vertebra level to expose spinal cord for contusion using weight-drop method. PBS-treated SCI rats (group one) were used as negative controls, in which cavitations were observed 10 weeks after SCI. pRNA-U6.1/Hygro- (group two, as a mock) and pRNA-U6.1/Hygro-p75 shRNA- (group three) transfected BMSC were labeled with a fluorescent dye, CM-DiI, and grafted into the lesion site 7 days after surgery. The Basso-Beattie-Bresnehan locomotor rating scale was performed weekly for 10 weeks. Results: There was a significant difference (P≤0.05) between all groups of treated rats regarding functional recovery. Specifically, the discrepancy among p75 siRNA and mock-transfected BMSC was statistically significant. P75 siRNA BMSC also revealed a higher level of in vivo survival compared to the mock BMSC. Conclusion: Our data suggest that genetically modified BMSC that express p75:siRNA could be a more suitable source of cells for treatment of SCI. Key Words: Spinal cord injury (SCI), Apoptosis, Bone marrow stromal cells (BMSC)     

Pharmacokinetics of Mirtazapine and Its Main Metabolites after Single Oral Administration in Fasting/Fed Horses

Mirtazapine (MRZ) is a human antidepressant drug that is metabolized, predominantly by the cytochrome P450 enzyme system, to 8-OH mirtazapine (8-OH MRZ) and dimetilmirtazapine (DMR) metabolites. In veterinary medicine, this drug is currently administered to cats and dogs with anorexia, although it could also have applications as an antidepressant, antiemetic, and analgesic agent in these species. The aim of this study was to assess the pharmacokinetics of MRZ and its metabolites DMR and 8-OH MRZ in horses. Six healthy female horses were administered MRZ (2 mg/kg) in fasting and fed states according to a balanced crossover study design. Plasma MRZ and metabolite concentrations were evaluated by high-performance liquid chromatography fluorescence detection method. Pharmacokinetic profiles of MRZ and DMR were similar (detected from 0.5 up to 34 and 48 hours, respectively), with an MRZ AUC_0-N/DMR AUC_0-N ratio range varying between 1.1 and 1.7. Surprisingly, 8-OH MRZ was undetected. Most of the pharmacokinetic parameters were not altered by food, with the exception of the time required to reach maximum concentration; this showed a statistical increase in subjects in the fasting state as compared with the fed state. However, because MRZ is an active substance intended for long-term administration, the slight increase of the time required to reach maximum concentration is not considered to be of any clinical consequence. In conclusion, the pharmacokinetic parameters demonstrated in this study suggest that MRZ is suitable for oral administration in the horse. However, further investigations are required to evaluate both its safety and effectiveness in this animal species.     

Preemptive analgesia with local lidocaine infiltration for single-level open disc operation

To evaluate the impact of preemptive local analgesia at the incision site for postoperative pain in patients undergoing disc operation. In this prospective, randomized, double-blinded, placebo-controlled study 166 patients were assigned to either lidocaine (n = 83) or placebo (n = 83) groups. The incision site was infiltrated with either 20 mL of 2% lidocaine and 0.9% saline in lidocaine group or 0.9% saline before the incision. Morphine (5 mg) was used for postoperative pain treatment. Postoperative pain was measured with Visual Analog Scale (VAS) in 6, 12, 24 and 48 h. Data were analyzed with SPSS software, using Chi-square and t-tests. The groups were matched for age, sex, type of operation, mean length of hospital stay and mean length of operation. Statistical analysis revealed no significant difference in visual analog scores of pain severity at 6, 12, 24 and 48 h after surgery between lidocaine and placebo groups (6 h: 38.22 +/- 26.87 vs. 34.52 +/- 24.43, p = 0.35; 12 h: 33.26 +/- 28.83 vs. 28.01 +/- 24.71, p = 0.20; 24 h: 26.71 +/- 23.31 vs. 22.85 +/- 22.48, p = 0.27; 48 h 16.35 +/- 10.16 vs. 15.23 +/- 8.90 p = 0.45). The amount of narcotics used post operatively had no meaningful difference in the groups (lidocaine 10.07 +/- 8.24 mg vs. placebo 10.54 +/- 9.31 mg p = 0.73). Preemptive analgesia with lidocaine 2% used subcutaneously before skin incision has no effect in reducing postoperative pain, narcotics demand and duration of hospital stay.     

Effect of Food Deprivation on Formalin-Induced Nociceptive Behaviors and Beta-Endorphin and Sex Hormones Concentration in Rats

Background: The present study examined the possible role of endogenous opioidergic system in effect of food deprivation on formalin-induced nociceptive behaviors in male and female rats. Also, we investigated the effect of food deprivation on the plasma level of beta-endorphin and sex hormones. Methods: Food was withdrawn 48 h prior to performing the formalin test, but water continued to be available ad libitum. The formalin was injected into hind plantar paw. Results: There is significant difference between male and female control rats during phase 2B. Following 48-h food deprivation, both male and female rats exhibited enhanced nociceptive behavior in response to formalin. Food deprivation for 12 and 24 h increased and for 48 h decreased beta-endorphin level in male and female rats. Food deprivation for 24 h decreased testosterone level in male, while it had no significant effect on female rats and food deprivation for 48 h decreased testosterone level in both sexes. Food deprivation for 24 h increased estradiol level in female and that for 48 h had no significant effect on male and female rats. Conclusions: The present study demonstrates the existence of food deprivation for 48 h causes enhancement of nociception in the formalin test in male and female rats that has correlation with decrease in plasma beta-endorphin and testosterone levels. Key Words: Rat, Food Deprivation, beta-endorphin, Naloxone     

In vitro antifungal activities of Allium cepa, Allium sativum and ketoconazole against some pathogenic yeasts and dermatophytes

By using an agar dilution assay, the antifungal activity of aqueous extracts prepared from Allium cepa (onion; AOE) and Allium sativum (garlic; AGE) were evaluated against Malassezia furfur (25 strains), Candida albicans (18 strains), other Candida sp. (12 strains) as well as 35 strains of various dermatophyte species and compared with the activity of a known antifungal drug, ketoconazole (KTZ). All the AOE, AGE and KTZ were found to be able to inhibit growth of all fungi tested in a dose-dependent manner with maximum of 100% at defined concentrations. The results indicate that onion and garlic might be promising in treatment of fungal-associated diseases from important pathogenic genera Candida, Malassezia and the dermatophytes.     

Forage quality, water use and nitrogen utilization efficiencies of pearl millet (Pennisetum americanum L.) grown under different soil moisture and nitrogen levels

The increasing scarcity of water for irrigation is becoming the most important problem for producing forage in all arid and semi-arid regions. Pearl millet is a key crop in these regions which needs relatively less water than other crops. In this research, a field study was conducted to identify the best combination of irrigation and nitrogen (N) management to achieve acceptable pearl millet forage both in quantity and quality aspects. Pearl millet was subjected to four irrigation treatments with interaction of N fertilizer (0, 75, 150 and 225 kg ha⁻¹). The irrigation treatments were 40%, 60%, 80% and 100% of total available soil water (I₄₀, I₆₀, I₈₀ and I₁₀₀, respectively). The results showed that increasing moisture stress (from I₄₀ to I₁₀₀) resulted in progressively less total dry matter (TDM), leaf area index (LAI), and nitrogen utilization efficiency (NUzE), while water use efficiency (WUE) and the percentage of crude protein (CP%) increased. The highest TDM and LAI were found to be 21.45 t ha⁻¹ and 8.65, in I₄₀ treatment, respectively. TDM, WUE, CP% and profit responses to N rates were positive. The maximum WUE of 4.19 kg DM/m³ was achieved at I₁₀₀ with 150 kg N ha⁻¹. The results of this research indicate that the maximum profit of forage production was obtained in plots which were fully irrigated (I₄₀) and received 225 kg N ha⁻¹. However, in the situation which water is often limited and not available, application of 150 kg N ha⁻¹ can produce high forage quality and guaranty acceptable benefits for farmers.