Animals: Forty SpragueDawleyrats, 20 males and 20 females, weighing 250-300gm were selected from the Animal Holding of the Faculty of Medical Sciences, University of the West Indies. Their fasting blood glucose levels were measured with One Touch Profile Glucose Meter (Johnson and Johnson Trinidad), before alloxan monohydrate (150mg/kg) was administered intraperitoneally to ten males and ten females. These represented the experimental animals. The rest were kept as control. Rats with blood glucose levels of 250- 600mg/dl were considered to be diabetic. The rats were maintained on the diabetic fasting glucose range with gliclazide (80mg/kg body weight) orally every 24
High fructose corn syrup (HFCS) is one of the most used sweeteners in the food industry. Health concerns regarding the consumption of HFCS-containing foods have developed in parallel with the increasing amount of people who become overweight. This study was conducted to investigate whether HFCS-containing peach nectar (pn-HFCS) consumption has more detrimental effects on anthropometrical and biochemical parameters compared with sucrose-containing peach nectar (pn-sucrose). Fifty-day-old SpragueDawleyrats were divided into three groups and were fed (A) pn-HFCS + ad libitum chow, (B) pn-sucrose + ad libitum chow and (C) only ad libitum chow for 7 months. The percentage change in body weight (PCBW), body mass index (BMI), and Lee index were calculated, and serum triglyceride, glucose, insulin and leptin concentrations were measured. The PCBW, BMI, Lee index, serum triglyceride, glucose, insulin and leptin concentrations were insignificant among the three groups. We can suggest that peach nectar consumption resulted in more energy intake than the control and since pn-HFCS group consumed more chow than the pn-sucrose group. The results show that long term daily HFCS or sucrose consumption in peach nectar is not associated with weight gain and does not stimulate metabolic changes in SpragueDawleyrats.
The Protein Digestibility Corrected Amino Acid Score (PDCAAS) of sweetpotato-based complementary foods (OFSP ComFa and CFSP ComFa) and cereal-based infant products (Weanimix and Cerelac) was assessed using 3 wk-old male SpragueDawleyrats weighing between 53–67 g as a model for human infants. Also, the effect of consumption of the infant formulations on lean mass, bone mass content and fat mass was evaluated by Dual-Energy X-ray Absorptiometry (DEXA) using 6 wk-old SpragueDawleyrats (initial weight, 206-229 g). The ComFa products and Weanimix are household-level formulations, and Cerelac is a commercial infant cereal. The true protein digestibility score for Cerelac was 96.27%, and about 1.8% (P<0.0001) higher than that for OFSP ComFa, CFSP ComFa and Weanimix. However, OFSP ComFa had the highest un-truncated PDCAAS by a difference of 4.1%, than CFSP ComFa, and about 20% difference compared with both the Weanimix and Cere- lac. All the products investigated had PDCAAS greater than 70%, the minimum protein qual- ity requirement for complementary foods. Among the rats assigned to the four formulations, their bone mass and fat mass composition were not significantly different (P=0.08 and P =0.85, respectively). However, the rats on CFSP ComFa had higher lean mass than those on Cerelac (321.67 vs. 297.19 g; P=0.03). The findings from the PDCAAS and the DEXA- measured body composition studies indicate that complementary foods could be formulated from readily available agricultural resources at the household-level to support growth as would a nutritionally adequate industrial-manufactured infant cereal. Nonetheless, it should be noted that the findings of our studies are based on an animal model.
Gene expression profiling is a tool to gain mechanistic understanding of adverse effects in response to compound exposure. However, little is known about how the common handling procedures of experimental animals during a preclinical study alter baseline gene expression. We report gene expression changes in the livers of female Sprague-Dawleyrats following common handling procedures. Baseline gene expression changes identified in this study provide insight on how these changes may affect interpretation of gene expression profiles following compound exposure. Rats were divided into three groups. One group was not subjected to handling procedures and served as controls for both handled groups. Animals in the other two groups were weighed, subjected to restraint in Broome restrainers, and administered water via oral gavage daily for 1 or 4 days with tail vein blood collections at 1, 2, 4, and 8 hours postdose on days 1 and 4. Significantly altered genes were identified in livers of animals following 1 or 4 days of handling when compared to the unhandled animals. Gene changes in animals handled for 4 days were similar to those handled for 1 day, suggesting a lack of habituation. The altered genes were primarily immune function related genes. These findings, along with a correlating increase in corticosterone levels suggest that common handling procedures may cause a minor immune system perturbance.
Alcohol consumption is associated with adverse effects on the cardiovascular and neural systems. Based on the emergent roles of the hormone system in mediating alcohol-induced anomalies, we were interested in understanding the link between alcoholism and estrogen signaling in the heart and brain tissue of Sprague-Dawleyrats. For that, we investigated the effect of chronic ethanol treatment on the stability of the expression levels of 15 housekeeping genes in rat heart and brain tissue for identifying most reliable genes as reference genes for gene expression analysis. To perform this study, we treated Sprague-Dawleyrats with ethanol (ETOH). The effect of ethanol on the stability of 15 reference gene candidates (Table 1) was investigated using rats with different hormonal backgrounds. The study was based on 6 rat groups. Untreated female (SHAM) and male rats were used as controls. One group of rats underwent ovariectomy (OVX). The last group of rats was ovariectomized and then treated with estrogen hormone (OVXE2). Rats belonging to those two groups were divided into two subgroups. Half of OVX as well as OVXE2 rats served as control (no ETOH treatment), while the remaining rats received ETOH.
In the present dissertation, we performed a longitudinal aging study in Sprague- Dawleyrats. Our samples consisted on males (n=42) and females (n=34) of two different genetic backgrounds, Wt (n=37) and Tg (Tg (CaMKII-hA2AR)) (n=39). We therefore describe the spectrum of spontaneous age-associated lesions that occur in these animals, and also perform statistical analysis using SPSS to evaluate the impact of the genotype of these rats in aging. Overall, Tg rats were 2.7 times more likely to develop pathology than Wt rats [OR 2.745, IC 95% 1.0.07-6.997 (p<0.05)]. When controlled the statistical analysis for age (i.e. age matched animals) Tg rats were 2.8 times more likely to develop age-associated lesions than Wt [ OR=2.8, (p>0.05)]. We also found that age was the most determinant factor for the occurrence of lesions and for every additional week of age the risk of onset of pathology increased by 1.060 times (OR 1.060, p<0.05). Since aging is defined as a time-dependent functional decline, progressive loss of physiological integrity and progressive increase in disease susceptibly (López-Otín, Blasco, Partridge, Serrano, & Kroemer, 2013; Franceschi & Campisi, 2014; Mitchell, Scheibye- Knudsen, Longo, & de Cabo, 2015), it was expected to have an increase of risk of onset of pathology for every extra week of age. Similar analysis was performed for cardiovascular, endocrine, pulmonary and renal pathology.
To date, the information with regard to effect of testosterone on ENaC expression in kidneys were far from complete. Quinkler et al. (2005) reported that sub-chronic (14 days) treatment of adult male Wistar rats with testosterone resulted in elevated α-Enac mRNA levels in kidney homogenates. Meanwhile, administration of dihydrotestosterone (DHT) causing the same effects, but was lesser than testosterone. Additionally, their study also showed that incubation of human kidney proximal tubule cell line with testosterone but not DHT caused increase in α-ENaC mRNA level with testosterone effect was being antagonized by flutamide. However, no expression for β and γ -ENaC was detected in these cells. In the meantime, testosterone and DHT were found to decrease expression of α, β and γ -ENaC subunits in ovariectomised female Wistar rat kidney (Kienitz et al., 2009). Based on these findings, we hypothesized that testosterone affects expression of all ENaC subunits i.e., α, β and γ in the kidneys, in which their co-existence will lead to a fully functioning ENaC channel, thus would augment Na +
Extract of Glycyrrhiza Glabra was dissolved in 0.05M NaOH at 37°C and pH was adjusted to 7.4 using 1M Hydrochloric Acid. For evaluation of the bleeding effect Wistar rats, were anaesthetized by using xylazin (16 mg/kg, intra muscularly) followed by ketamin, (100 mg/kg)
NF-κB is a cardinal transcriptional factor that involved in the response of inﬂammation and cancers 25 . Moreover, more studies have been done on NF-κB related to chronic pain. For instance, recent studies have shown its critical role in chemotherapy-induced chronic pain 26 . We were specifically interested the molecular mechanism in LDH induced pain. Interestingly, NF-κB was activated in LDH rats and its activation was regulated by the expression of CX3XL1. For further studies, as it is well known that Akt, which is activated by phosphatidylinositol 3-kinase (PI3K), could phosphorylates NF-κB’s inhibitor κB (IκB). This phosphorylation could leads to ubiquitination and degradation of IκB; thus leading to the activation of NF-κB 27 . However, how this
knockout mice and AT2R or MrgD-transfected CHO cells. Materials and methods: The endothelium-dependent vasodilatory response to Ang-(1-9) was tested in aortic rings taken from Wild-Type, AT2KO and MASKO mice and Sprague-Dawleyrats, pre-contracted with phenylephrine (0.1 umoles/L). NO release from AT2R or MrgD stable transfected CHO cells was evaluated using the NO indicator 4-amino-5 methylamino-2, 7 difluorofluoresceindiacetate (DAF- FM) after Ang-(1-9) stimulation. Analyzed the in vivo cardiovascular parameters were recorded by a signal sent to a transducer connected to the cannula inserted into the abdominal aorta through the femoral artery, through data acquisition system (Biopac System, model MP150).Results: In aortic rings from SD rats Ang-(1-9) produced a dose-related relaxation which was not modified by A-779, the Mas/MrgD antagonist D-Pro7-Ang-(1-7); or by PD123319. In aortic rings taken from Mas KO there was a minor attenuation of the Ang-(1-9) vasorelaxant effect when compared to the WT. No difference between AT 2 KO
Introduction: The male reproductive system is subject of studies to be sensitive to changes in androgens concentration, and these are changed in some situations such as exercise, use of anabolic steroids (AAS) and age. One of androgen-dependent organs is the epididymis. Objective: To determine if the treatment with AAS associated or not to exercise, in Sprague-Dawleyrats alters the morphology of the epididymis in adult rats and its chronic effects in the elderly. Methods: It was analyze epididymis of 56 Sprague-Dawleyrats, virgins, with 13 weeks old; they were divided into eight groups with seven animals each: GC - adults and sedentary; GCi - elderly and sedentary; GN - adults, sedentary treated with AAS; GNi - elderly, sedentary treated with AAS; GE - adults treated with exercise; GEi - elderly treated with exercise; GNE - adults, exercise and treatment with AAS; GNEi - elderly, exercise and treatment with AAS. The training was conducted by jumping in the water with overload. The AAS administration occurred by intramuscular injection of nandrolone decanoate (10 mg/kg/week). Epididymal samples passed by histological routine of hematoxylin and eosin for morphological and morphometric analysis. Results: The duct diameters was lower when compared GE to other groups, and GN and GNE, had diameter and epithelial height increased compared to GC. The aged groups tended to return to normal. There was also inflammatory signs in the tissue of the experimental groups (GN, GE, GNE and GCI).
Ovariectomized Balb/C mice treated with raloxifene analogs and fed with fish oil showed a reduction in the production of the inflammatory cytokines IL-1b and IL-6 . In fact, during the inflammatory response there are released pro-proliferative cytokines that can stimulate the tumor growth. Inflammatory response is part of the innate immune response. Innate immune cells present in the tumors may have a dual effect: although tumor-associated macrophages may kill tumor cells, they are able to produce angiogenic and lymphangiogenic growth factors, cytokines and proteases that can potentiate the neoplastic progression and blunt anti-tumor response of T cytotoxic cells . By analyzing a cDNA microarray of tumors of corn oil-fed (an omega-6-rich oil) + tamoxifen-treated rats we found that these animals had increased expression of genes related to the increment of the inflammatory response, that was not found in fish oil+TAM-treated animals . Additionally, the treatment of Sprague-Dawleyrats with fish oil increased the modulation of genes that indicate a better immune response against tumors (polarized Th1 immune response) .
Wake-sleep (W-S) states are affected by thermoregulation. In particular, REM sleep (REMS) is reduced in homeotherms under a thermal load, due to an impairment of hypothalamic regulation of body temperature. The aim of this work was to assess whether osmoregulation, which is regulated at a hypothalamic level, but, unlike thermoregulation, is maintained across the different W-S states, could influence W-S occurrence. Sprague-Dawleyrats, kept at an ambient temperature of 24uC and under a 12 h:12 h light-dark cycle, were exposed to a prolonged osmotic challenge of three days of water deprivation (WD) and two days of recovery in which free access to water was restored. Two sets of parameters were determined in order to assess: i) the maintenance of osmotic homeostasis (water and food consumption; changes in body weight and fluid composition); ii) the effects of the osmotic challenge on behavioral states (hypothalamic temperature (Thy), motor activity, and W-S states). The first set of parameters changed in WD as expected and control levels were restored on the second day of recovery, with the exception of urinary Ca ++ that almost disappeared in WD, and increased to a high level in recovery. As
MATERIALS AND METHODS The present study was conducted on 24 healthy male Spraguedawleyrats in two groups where 12 animals were for contextual fear condition and another 12 were for modified step-down method of learning and memory. The rats were procured and housed in cages at department of veterinary pathology, West Bengal University of Animal and Fishery Sciences, Kolkata, India. Three animals per cage were accommodated in polycarbonate cages during the experimental period. Husk was used as bedding material. Husk was strained and sterilized by autoclaving in transparent polythene bag using sterol strip as an indicator of successful sterilization. Changing frequency of bedding material was twice a week. Animals were allowed to acclimatize for a period of 7 days prior to experiment and provided standard feed (Nutri Lab, rodent feed, Vetcare Pvt. Ltd, Bangalore) and allowed water ad libitum.
We assessed the effects of lodenafil on hemodynamics and inflammation in the rat model of monocrotaline- induced pulmonary hypertension (PH). Thirty male Sprague-Dawleyrats were randomly divided into three groups: control; monocrotaline (experimental model); and lodenafila (experimental model followed by lodenafil treatment, p.o., 5 mg/kg daily for 28 days) Mean pulmonary artery pressure (mPAP) was obtained by right heart catheterization. We investigated right ventricular hypertrophy (RVH) and IL-1 levels in lung fragments. The number of cases of RVH was significantly higher in the monocrotaline group than in the lodenafil and control groups, as were mPAP and IL-1 levels. We conclude that lodenafil can prevent monocrotaline-induced PH, RVH, and inflammation.
We assessed the effects of lodenafil on hemodynamics and inflammation in the rat model of monocrotaline- induced pulmonary hypertension (PH). Thirty male Sprague-Dawleyrats were randomly divided into three groups: control; monocrotaline (experimental model); and lodenafil (experimental model followed by lodenafil treatment, p.o., 5 mg/kg daily for 28 days) Mean pulmonary artery pressure (mPAP) was obtained by right heart catheterization. We investigated right ventricular hypertrophy (RVH) and IL-1 levels in lung fragments. The number of cases of RVH was significantly higher in the monocrotaline group than in the lodenafil and control groups, as were mPAP and IL-1 levels. We conclude that lodenafil can prevent monocrotaline-induced PH, RVH, and inflammation.
Twenty adult male Sprague-Dawleyrats weighing 230- 250 g were used in this study. All rats were obtained from the Laboratory Animal Resource Unit, Faculty of Medicine, Universiti Kebangsaan Malaysia, one week prior to the study for acclimatization purposes. The rats were kept in plastic cages, with two rats per cage under standard environmental conditions (12 h light/dark cycles, 25-28 ˚ C) and maintained on a standard pellet and water ad libitum diet for the duration of the study. All of the animal handling protocols were approved by the Animal Ethics Committee of Laboratory Animal Resource Unit, Faculty of Medicine, Universiti Kebangsaan Malaysia.
Experiments, approved by the Animal Welfare Ethics Committee of Flinders University, were carried out on 37 male Sprague– Dawleyrats (300–400 g). Care was taken to minimize the number of animals. For implantation of measuring devices rats were anaesthetized with 2% isoﬂurane (Veterinary Companies of Australia Pty Ltd., NSW, Australia) in 100% oxygen. An ultrasonic Doppler blood ﬂow probe (Iowa Doppler Products, IA, USA) was implanted around the tail artery about 2 cm distal to the base (Garcia et al., 2001; Ootsuka et al., 2009). After cannula implantation for amygdala injections (see below) ﬂow probes were connected via subcutaneous wires to a head- piece attached to the skull.
Finally, the fact should be addressed that differences in call emission cannot only be observed within, but also between defined subject samples. For one, gender differ- ences were reported but these differed between studies and tests: Graham et al. (72) observed more 22-kHz calls (and more freezing) during fear conditioning in males, whereas Blanchard et al. (41) observed more calls in females kept in a social environment and with a cat as the threat stimulus. With respect to specific breeding lines, Naito et al. (73) studied Tsukuba high- versus low-emotional rats, which are derived from Wistar rats, differing in locomotor activity or freezing in a light/dark runway. They found that the percent- age of rats emitting 22-kHz calls as an acute response to electric footshock was higher in high- than low-emotional rats (68 versus 47%). Also, they found that activation of the hypothalamic-pituitary-adrenal (HPA) axis, i.e., adre- nocorticotropic hormone (ACTH) and corticosterone, did not differ between vocalizing and non-vocalizing rats, that is, the differences in call emission cannot be explained in terms of the acute stress responses induced. Finally, com- mon outbred rat strains were compared, and Walker et al. (74), using a resident/intruder paradigm, found more calls in Sprague-Dawley than Wistar rats, which also showed less submissive postures. Sprague-Dawleyrats were also found to emit more 22-kHz calls during fear conditioning compared to Long-Evans rats (72), or compared to Long- Evans and Wistar rats (33). Importantly, the differences in calling were not paralleled by similar ones in freezing, with Long-Evans rats showing the highest rates (33). The call differences between strains may be explained by differences in emotionality, coping styles, or defense mechanism, or by differences in aversive learning in these conditioning pro- cedures. Interestingly, the adult calling patterns in 22-kHz calls were not paralleled by similar ones in 40-kHz calls of pups during isolation, where Sprague-Dawleyrats showed the lowest rates (33), again supporting our hypothesis that pup vocalization should not be taken as a general read-out of anxiety.
Neonatal Sprague-Dawleyrats were randomly divided into normal control, mild hypoxia-ischemia (HI), and severe HI groups (N = 10 in each group at each time) on postnatal day 7 (P7) to study the effect of mild and severe HI on anxiety-like behavior and the expression of tyrosine hydroxylase (TH) in the substantia nigra (SN). The mild and severe HI groups were exposed to hypoxia (8% O 2 /92% N 2 ) for 90 and 150 min, respectively. The elevated plus-maze (EPM) test was performed to assess anxiety-like behavior by measuring time spent in the open arms (OAT) and OAT%, and immunohistochemistry was used to determine the expression of TH in the SN at P14, P21, and P28. OAT and OAT% in the EPM were significantly increased in both the mild (1.88-, 1.99-, and 2.04-fold, and 1.94-, 1.51-, and 1.46-fold) and severe HI groups (1.69-, 1.68-, and 1.87-fold, and 1.83-, 1.43-, and 1.39-fold, respectively; P < 0.05). The percent of TH-positive cells occupying the SN area was significantly and similarly decreased in both the mild (17.7, 40.2, and 47.2%) and severe HI groups (16.3, 32.2, and 43.8%, respectively; P < 0.05). The decrease in the number of TH-positive cells in the SN and the level of protein expression were closely associated (Pearson correlation analysis: r = 0.991, P = 0.000 in the mild HI group and r = 0.974, P = 0.000 in the severe HI group) with the impaired anxiety-like behaviors. We conclude that neonatal HI results in decreased anxiety-like behavior during the juvenile period of Sprague-Dawleyrats, which is associated with the decreased activity of TH in the SN. The impairment of anxiety and the expression of TH are not likely to be dependent on the severity of HI.