Saline IP Research Articles

Immunostimulatory and anti-inflammatory impact of Fragaria ananassa methanol extract in a rat model of cadmium chloride-induced pulmonary toxicity.

Cadmium is an extremely dangerous heavy metal that can lead to disastrous consequences in all organisms. Several natural remedies reduce the toxicities of experimentally generated metals in animals. Strawberry Fragaria ananassa contains several bioactive compounds that may mitigate heavy-metal toxicity. The study aim was to evaluate the ability of a strawberry fruit methanol extract (SE) to reduce Cd toxicity and to identify and quantify the active constituents of SE. Forty Wistar rats were classified into four groups: the control group- 1 ml saline IP; SE group- 100 mg of SE/kg rats orally; cadmium (Cd) group-2 mg CdCl2/kg body weight/IP daily; and treated group- SE given 1 hour before Cd administration. Administration of Cd induced several histopathological and immunohistochemical alterations in lung sections. Biochemical analysis of lung homogenates and mRNA levels of antioxidants and inflammatory cytokines indicated significant changes to the risk profile. SE administration significantly decreased the oxidative stress, inflammation, tissue damage, the mean area percentage of collagen fibers, and positive immuno-expressions of TNF-α and NF-κB induced by CdCl2. Moreover, the MDA, TNF-α, GM-CSF, and IL-1β levels in Cd-exposed rat lung tissue were significantly lower in the SE-treated group than in the Cd-group. SE significantly augmented lung GSH, SOD, HO-1, GPx-2, and Nrf2 levels in Cd-exposed rats. SE mitigated Cd-caused oxidative stress and lung inflammation. Therefore, regularly consuming a strawberry-rich diet could benefit general health and help prevent and treat diseases.

Carnosine and crocin ameliorate oxidative stress in rats with rhabdomyolysis-induced acute kidney injury through upregulating HO-1 gene expression

We aimed to investigate whether carnosine (a natural dipeptide found in humans) and crocin (a natural product extracted from saffron and gardenia) can prevent the harmful effects of acute kidney injury (AKI) caused by glycerol-induced rhabdomyolysis with a focus on the role of the nuclear factor E2-related factor-2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway. Thirty-six male Wistar rats were divided into 6 groups: 3 of them received saline, carnosine, and crocin IP for 7 days and the other 3 groups received the same treatment in addition to a single IM injection of 50% glycerol (10 ml/kg, divided between the two limb muscles) on 8th day. All animals were sacrificed after 24 h of the IM injection. Blood samples, muscles, and kidneys were collected for further investigations. Rhabdomyolysis was evidenced by the histopathological alterations in muscle sections and increased levels of myoglobin in kidney tissue homogenate samples. Kidney injury was characterized by increased creatinine, kidney injury molecule-1, malondialdehyde, nitric oxide, and decreased catalase activity. The injury also increased inflammatory markers and histopathological alterations in kidney sections. All these effects were corrected by pretreatment with carnosine and crocin. Both agents could also elevate HO-1 gene expression. However, both agents failed to restore the declined Nrf2 expression in glycerol-treated groups. Carnosine and crocin can effectively prevent rhabdomyolysis-induced AKI in rats through augmenting gene expression of HO-1 and antioxidant system and suppressing the generation of reactive oxygen species, lipid peroxidation, and inflammatory response.

Swallowing Kinematics in Rats Treated with Low Dose Chemotherapy Concurrently with Fractionated Radiation

Dysphagia is a common consequence of chemotherapy and radiation treatment for oropharyngeal cancer. Injury is thought to occur due to damage to the muscle and/or nerves. This can severely impair swallowing function, leading to serious negative outcomes such as aspiration related pneumonia or tube feeding placement to maintain nutrition. Currently, there are only strategies to prevent swallowing dysfunction; however, the development of chronic radiation associated dysphagia persists effecting ~25% of head and neck cancer survivors. Therefore, there is a need to study the mechanisms associated with radiation associated dysphagia to help guide new treatments to improve swallowing outcomes. The present study investigated the effects of concurrent chemoradiation on swallowing function. We tested the hypothesis that fractionated radiation with low doses of chemotherapy alters swallowing kinematics and bolus transit compared to radiation alone or normal controls. We monitored the functional extent of chemoradiation or radiation treatment using videofluoroscopy swallow study in self feeding rats (n=16) at four weeks post treatment. Twelve rats underwent 48Gy of fractionated radiation to the submental muscles using a clinical linear accelerator given in 12 fractions of 4Gy. Irradiated rats received either cisplatin 1mg/kg or saline IP once per week for four weeks during radiation treatment. Four rats received no treatment. During videofluoroscopy, thin liquids and puree consistencies mixed with 40% w/v barium sulfate were given ad libitum. The movement trajectories of the tongue, mandible, and hyoid bone were quantitatively tracked from video data. Changes in the speed of the jaw and hyoid during swallowing were observed between treated and control rats. No differences were observed in bolus flow parameters i.e. pharyngeal transit speed, inter‐swallow interval, and swallow frequency between treatment groups. The average size of the bolus in vallecula decreased by ~4% prior to initiating pharyngeal swallow with chemoradiated rats (p < 0.05). Results suggest that low doses of chemotherapy in conjunction with radiation appear to have minimal differences in swallowing kinematics compared to radiation alone. Higher doses of chemotherapy may be needed to induce similar abnormalities in swallowing as seen clinically.

The effect of vitamin E on the histological structure of kidney in rats treated with cyclophosphamide

Cyclophosphamide (CP) is a cytotoxic alkylating drug that is commonly used to treat autoimmune disorders and cancer. CP causes nephrotoxicity by producing highly reactive radicals devoid of oxygen. A fat-soluble vitamin E that interacts as a free radical scavenger, inhibiting nitrosamine production and blocking lipid peroxidation from polyunsaturated fatty acids. The aim of this research was to see if vitamin E may help combating CP induced nephrotoxicity. Thirty-two adult albino female Westar rats were randomly divided into four groups (n=8) and received daily treatment for thirty days. Group A considered as control group, receiving normal saline IP at dose of 0.3 ml, group B given 15 mg/kg b.w. of cyclophosphamide IP, while group C received cyclophosphamide with vitamin E at dose 200 mg/kg, oral administration of vitamin E one hour before injection of cyclophosphamide, in group D they received vitamin E orally at dose of 200 mg kg body weight mixed with olive oil daily by oral gavage. In group B the cyclophosphamide causes highly significant reduction in body weight and histologically showed atrophy of most glomeruli, distention in bowman’s space, tubular degeneration and luminal hyaline cast material, while in group C showed most of the glomeruli normal except atrophy of few number of glomeruli and focal epithelial degeneration of renal tubules. Our study found that one of the primary mechanisms induced by cyclophosphamide may be nephrotoxicity due to oxidative stress, and pretreatment with vitamin E reduces cyclophosphamide-induced nephrotoxicity.

Ayahuasca, a psychedelic beverage, modulates neuroplasticity induced by ethanol in mice

Alcohol use disorder needs more effective treatments because relapse rates remain high. Psychedelics, such as ayahuasca, have been used to treat substance use disorders. Our study aimed to evaluate the effects of ayahuasca on ethanol-induced behavioral sensitization (EIBS). Swiss mice received 2.2 g/kg ethanol or saline IP injections every other day across nine days (D1, D3, D5, D7, and D9), and locomotor activity was evaluated 10 min after each injection. Then, animals were treated daily with ayahuasca (corresponding to 1.76 mg/kg of N,N-dimethyltryptamine, DMT) or water by oral gavage for eight consecutive days. On the seventh day, mice were evaluated in the elevated plus maze. Then, mice were challenged with a single dose of ethanol to measure their locomotor activity. Dopamine receptors, serotonin receptors, dynorphin, and prodynorphin levels were quantified in the striatum and hippocampus by blot analysis. Repeated ethanol administration resulted in EIBS. However, those animals treated with ayahuasca had an attenuated EIBS. Moreover, ayahuasca reduced the anxiogenic response to ethanol withdrawal and prevented the ethanol-induced changes on 5-HT1a receptor and prodynorphin levels in the hippocampus and reduced ethanol effects in the dynorphin/prodynorphin ratio levels in the striatum. These results suggest a potential application of ayahuasca to modulate the neuroplastic changes induced by ethanol.

Neuroprotective effect of paricalcitol in a rat model of transient global cerebral ischemia

BackgroundParicalcitol is known to attenuate ischemic-reperfusion injury of various organs. However, it is not known whether paricalcitol prevents neuronal injury after global cerebral ischemia. The purpose of this study is to investigate the neuroprotective effect of paricalcitol in a rat model of transient global cerebral ischemia.MethodsThis is a prospective, randomized experimental study. Male Sprague-Dawley rats that survived 10 min of four-vessel occlusion were randomly assigned to two treatment groups: one group was treated with paricalcitol 1 μg/kg IP, and the other was given an equivalent volume of normal saline IP. Drugs were administered at 5 min, 1 day, 2 days, and 3 days after ischemia. Neurologic function was assessed at 2 h, 1 day, 2 days, 3 days, and 4 days after ischemia. We tested motor function 3 days after ischemia using the rotarod test. Also, we tested memory function 4 days after ischemia using the passive avoidance test. We assessed neuronal degeneration in the hippocampus of surviving rats 4 days after ischemia.ResultsEight rats were allocated to each group. No significant differences were found between the groups in terms of survival rate, motor coordination, or memory function. The neurological function score 2-h post-ischemia was significantly higher in the paricalcitol group (p = 0.04). Neuronal degeneration was significantly less in the paricalcitol group compared with the control group (p = 0.01).ConclusionsParicalcitol significantly attenuated neuronal injury in the hippocampus. Although motor coordination, memory function, and survival rate were not significantly improved by paricalcitol treatment in this study, paricalcitol remains a potential neuroprotective drug after global cerebral ischemia.

Vigabatrin-Induced Retinal Functional Alterations and Second-Order Neuron Plasticity in C57BL/6J Mice.

PurposeVigabatrin (VGB) is an effective antiepileptic that increases concentrations of inhibitory γ-aminobutyric acid (GABA) by inhibiting GABA transaminase. Reports of VGB-associated visual field loss limit its clinical usefulness, and retinal toxicity studies in laboratory animals have yielded conflicting results.MethodsWe examined the functional and morphologic effects of VGB in C57BL/6J mice that received either VGB or saline IP from 10 to 18 weeks of age. Retinal structure and function were assessed in vivo by optical coherence tomography (OCT), ERG, and optomotor response. After euthanasia, retinas were processed for immunohistochemistry, and retinal GABA, and VGB quantified by mass spectrometry.ResultsNo significant differences in visual acuity or total retinal thickness were identified between groups by optomotor response or optical coherence tomography, respectively. After 4 weeks of VGB treatment, ERG b-wave amplitude was enhanced, and amplitudes of oscillatory potentials were reduced. Dramatic rod and cone bipolar and horizontal cell remodeling, with extension of dendrites into the outer nuclear layer, was observed in retinas of VGB-treated mice. VGB treatment resulted in a mean 3.3-fold increase in retinal GABA concentration relative to controls and retinal VGB concentrations that were 20-fold greater than brain.ConclusionsNo evidence of significant retinal thinning or ERG a- or b-wave deficits were apparent, although we describe significant alterations in ERG b-wave and oscillatory potentials and in retinal cell morphology in VGB-treated C57BL/6J mice. The dramatic concentration of VGB in retina relative to the target tissue (brain), with a corresponding increase in retinal GABA, offers insight into the pathophysiology of VGB-associated visual field loss.

Small molecule inhibition of protein phosphatase 2A in vivo does not disrupt glucose homeostasis in mice.

Identifying new targets for therapeutic intervention that might attenuate cardiovascular complications associated with type 2 diabetes is needed. Earlier we reported that mice treated with a potent and selective small molecule inhibitor (LB1; Lixte Biotechnology Holdings, Inc., East Setauket, NY; 1 mg/kg IP) of protein phosphatase 2A (PP2A) for the last 14 days of an obesogenic diet (12 weeks of fat‐feeding) did not display vascular PP2A hyperactivation, endothelial dysfunction, or hypertension that otherwise developed in fat‐fed mice treated with vehicle (Bharath et al, Diabetes, 2015). We concluded that PP2A hyperactivation should be considered further as a therapeutic target for intervention. Results from another laboratory group indicated that 2 mg/kg LB1 IP (3 hours) evoked hyperglycemia, glucose intolerance, and hepatic glycogen depletion in chow‐fed and fat‐fed (3 days) rats. The authors concluded that targeting PP2A hyperactivation should not be considered as a therapeutic intervention. Based on the incongruent conclusions from the two studies we tested the hypotheses that LB1 treatment for 14‐days impairs: (i) peripheral glucose homeostasis; (ii) hepatic glycogen content; and (iii) insulin‐mediated signal transduction in the liver. To test the first two hypotheses, 7‐week old male C57BL/6J mice consumed standard (Con) or high fat (HF) chow for 12 weeks. Subgroups of Con and HF mice received 1 mg/kg LB1 or saline (Veh) IP for the last 14 days. First, glucose and insulin tolerance testing indicated that LB1 treatment does not impair peripheral glucose homeostasis in Con mice or HF mice. Second, LB1 treatment did not lower hepatic glycogen content, p‐Akt2S474/Akt2, p‐Gsk3ɑS21/Gsk3ɑβ, or p‐GSS641/GS, in lean or obese mice under basal, random‐fed conditions. To test the third hypothesis, age‐matched male mice that consumed standard chow were treated with LB1 or Veh for 14 days. On day 15, after a 6 h fast, insulin or saline was administered (i.v.) to anesthetized mice, and segments of liver were collected to assess p‐Akt2S474/Akt2, p‐GSK3ɑS21/GSK3ɑ, and p‐GSS641/GS. Insulin‐mediated signal transduction in the liver was similar regardless of LB1 treatment. We conclude that LB1 treatment in mice for 14‐days does not impair: (i) peripheral glucose homeostasis; (ii) hepatic glycogen content; or (iii) insulin‐mediated signal transduction in the liver.Support or Funding InformationAmerican Diabetes Association (ADA: 1‐12‐BS‐208, ADA 7‐08‐RA‐164) and National Institutes of Health (NIH: 2R15HL091493) to JDSThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Evaluation of interobserver agreement for postoperative pain and sedation assessment in cats.

OBJECTIVE To evaluate agreement between observers with different training and experience for assessment of postoperative pain and sedation in cats by use of a dynamic and interactive visual analog scale (DIVAS) and for assessment of postoperative pain in the same cats with a multidimensional composite pain scale (MCPS). DESIGN Randomized, controlled, blinded study. ANIMALS 45 adult cats undergoing ovariohysterectomy. PROCEDURES Cats received 1 of 3 preoperative treatments: bupivacaine, IP; meloxicam, SC with saline (0.9% NaCl) solution, IP, (positive control); or saline solution only, IP (negative control). All cats received premedication with buprenorphine prior to general anesthesia. An experienced observer (observer 1; male; native language, Spanish) used scales in English, and an inexperienced observer (observer 2; female; native language, French) used scales in French to assess signs of sedation and pain. Rescue analgesia was administered according to MCPS scoring by observer 1. Mean pain and sedation scores per treatment and time point, proportions of cats in each group with MCPS scores necessitating rescue analgesia, and mean MCPS scores assigned at the time of rescue analgesia were compared between observers. Agreement was assessed by intraclass correlation coefficient determination. Percentage disagreement between observers on the need for rescue analgesia was calculated. RESULTS Interobserver agreements for pain scores were good, and that for sedation scores was fair. On the basis of observer 1's MCPS scores, a greater proportion of cats in the negative control group received rescue analgesia than in the bupivacaine or positive control groups. Scores from observer 2 indicated a greater proportion of cats in the negative control group than in the positive control group required rescue analgesia but identified no significant difference between the negative control and bupivacaine groups for this variable. Overall, disagreement regarding need for rescue analgesia was identified for 22 of 360 (6.1%) paired observations. CONCLUSIONS AND CLINICAL RELEVANCE Interobserver differences in assessing pain can lead to different conclusions regarding treatment effectiveness.

Ceftriaxone reduces alcohol intake in outbred rats while upregulating xCT in the nucleus accumbens core

Alcohol addiction is a chronic disease characterized by an inability to regulate drinking. A critical brain region involved in alcohol consumption is the nucleus accumbens (NA). Glutamate transmission in this region regulates alcohol consumption and relapse to alcohol-seeking. Across multiple alcohol-administration rodent models, basal extracellular glutamate levels are increased in the NA during early withdrawal. Glutamate transporter 1 (GLT-1) and system xC−, containing the subunit xCT, regulate NA glutamate levels. Ceftriaxone (Cef) increases expression and function of both transporters following extinction from cocaine self-administration and here we sought to determine if Cef would similarly decrease alcohol consumption while increasing xCT and GLT-1 in the NA core. We used the intermittent access to alcohol (IAA) paradigm to induce drinking in outbred Sprague-Dawley rats; this paradigm permits rats access to alcohol (20%v/v) for 24-h without water deprivation, followed by 24-h of abstinence. Following 17 24-h drinking sessions, Cef treatment (200mg/kg IP) was initiated and continued for 5days while a control group received vehicle (0.9% saline IP). Alcohol consumption was assessed for two 24-h periods during Cef and two 24-h periods after cessation of Cef treatment. In a separate cohort of rats, Cef's ability to alter blood alcohol levels (BALs) after a non-contingent alcohol injection (1g/kg) was assessed. We found that Cef decreased alcohol consumption during the period of Cef treatment and on the two days following injections, and this was accompanied by an increase in NA core xCT expression. Furthermore, a history of alcohol consumption did not alter xCT and GLT-1 expression relative to alcohol-naïve controls. Cef did not alter BALs, indicating that the reduction in alcohol consumption was not caused by altered alcohol clearance. These results indicate that while Cef reduces alcohol consumption in outbred rats, its ability to do so is not associated with an increase in GLT-1 expression.

Effects of Zinc Supplementation on Cardiac Remodeling After Experimental Myocardial Infarction

ObjectiveTo analyze the effects of zinc supplementation on cardiac remodeling after myocardial infarction. Methods: After experimental infarction, Wistar rats were allocated in two groups: Infarction Control (IC, n=23), that received saline IP every other day, and Infarction Zinc (IZ, n=24), with zinc supplementation (0.5mg/kilo) IP every other day. Sham animals were included and divided in two groups: Sham Control (SC; n=15), treated with saline, and Sham Zinc (SZ; n=15), with zinc supplementation. After 3 months, the animals were submitted to echocardiogram. The effect of infarction (I), Zinc (Z) and the interaction between the factors (IxZ) were analyzed by two way ANOVA. Results and conclusion: Zinc attenuated remodeling by reducing systolic area (SC: 22.6 ± 3.7; SZ: 11.2 ± 3.7; IC: 50.1 ± 2.6; IZ: 44 ± 2.6; p(I) <0.001, p(Z) 0.007, p(IxZ) 0.401) and diastolic area (SC: 50.8 ± 4.7; SZ: 40.1 ± 4.7; IC:79.4 ± 3.2; IZ: 74.9 ± 3.2; p(I) <0.001, p(Z) 0.057, p (IxZ) 0.43) and improving systolic function (shortening velocity of posterior wall SC: 55.6 ± 2.5; SZ: 64 ± 2.5; IC: 36 ± 1.6; IZ: 39.2 ± 1.6; p(I) <0.001, p(Z) 0.01, p(IxZ) 0.245) and diastolic function (E/E'= SC: 13.2 ± 1; SZ: 11.9 ± 1; IC: 17.6 ± 0.7; IZ: 15.5 ± 0.7; p(I) <0.001, p(Z) 0.052, p(IxZ) 0.598). Financial support: Fapesp: 13/06862‐4; 12/14500‐2.

A study of the effect of Nigella Sativa (black seeds) on Methotrexate induced hepatotoxicity in rabbits.

Background: To evaluate the effectiveness of Nigella sativa (NS) in the prevention of hepatotoxicity of large doses of methotrexate (MTX) (IP) in rabbits. Methods: Three groups of male rabbits, six in each were used. Oral dosing was administered as a paste; formula 1 was prepared by mixing 2 g flour with water; formula 2 contained flour and NS and water. Group 1 was fed with formula 1 daily and injected with 2 ml/kg normal saline IP. Group 2 was given formula 1 daily with 20 mg/kg MTX IP. Group 3 was fed with formula 2 daily + MTX 20 mg/kg IP. Injections were given weekly for 5 weeks, and then the animals were sacrificed at day 39. Liver enzymes, malondialdehyde (MDA), glutathione (GSH), and histopathology of the liver were evaluated. Results: Liver enzymes, serum, and liver MDA were significantly increased by MTX. MTX + NS treatment significantly reduced the rise in liver enzymes, MDA in serum with little effect on liver MDA. Serum aspartate aminotransferase, alkaline phosphatase, and bilirubin were reduced from 82.8±18.04 U/L, 4.9±2.0 kind and king unit/100 ml and 0.74±0.1 mg/dl to 56.1±7.5, 2.0±0.6 and 0.27±0.1 respectively. Unexpectedly, serum and liver GSH were slightly increased by MTX. Treatment with MTX + NS further increased these levels. Histologically, portal and lobular sinusoidal dilatation, lymphocytic infiltration, and hepatocyte hydropic degeneration were seen in all rabbits on MTX, which disappeared in three rabbits on NS + MTX. Conclusion: NS is hepatoprotective against MTX induced hepatotoxicity.

Effects of valproic acid and dexamethazone in acute kidney ischemia-reperfusion injury model (P2216)

Abstract Introduction: Renal ischemia-reperfusion (IR) causes acute kidney injury (AKI) with high mortality and morbidity. The objective of this study was to ameliorate kidney IR injury and identify novel biomarkers for AKI. Methods: Left renal ischemia was induced in Wister rats by clamping the renal artery for 45 minutes, followed by reperfusion and right nephrectomy. Thirty minutes prior to ischemia, rats (n=8/group) received Valproic Acid (150 mg/kg; VPA), Dexamethasone (3 mg/kg; Dex) or Vehicle (Saline) IP. Animals were sacrificed at 3h, 24h or 120h IR and blood, urine and kidney were collected. Results: At 3h IR, urine albumin was lower (P>0.05) in VPA (0.84±0.62) and Dex (1.04±0.73) compared to Vehicle (1.47±1.02 mg/ml) group; and Lipocalin-2 was lower (P<0.006) in Dex (0.42±0.15) and higher (P<0.01) in VPA (1.21±0.32) compared to Vehicle (0.77±0.31 µg/ml) group. Osteopontin was higher (P<0.05) at 24h IR in VPA and Dex . TIM-1 did not differ. Serum creatinine (mg/dL) at 24 h IR in VPA (2.7±1.8) and Dex (2.3±1.2) was reduced (P<0.05) compared to Vehicle (3.8±0.5). Histopathology at 3h IR demonstrated reduced (P<0.04) ischemic changes in the renal cortex in VPA (27± 32 %) compared to Vehicle (56±27%) group. Tubular necrosis in the outer medulla was lower (P<0.02) in Dex and VPA (43-45%) compared to Vehicle (70%) at 24h IR. BCL-2 as determined by RT-PCR did not differ. Conclusion: The VPA administration prior to surgery appears to offer renal protection from IR injury.

Ibuprofen does not reverse time‐dependent increase in hypoxic ventilation in chronically hypoxic rats

Chronic sustained hypoxia (CSH) increases inflammatory cytokine gene expression in carotid bodies and medullary respiratory centers, and increases carotid body chemoreceptor and ventilatory responses to hypoxia in rats. Non‐steroidal anti‐inflammatory drugs (ibuprofen IBU) block these responses (Respir.Physiol.Neurobiol. (2011) 178: 362–369 & 381–386). We tested the hypothesis that IBU can reverse these responses after they are already established in CSH. We measured cytokine expression in medullary respiratory centers and ventilatory responses in adult male rats acclimatized to CSH (PiO2 = 70 Torr, 1 wk) before and after IBU (4 mg/kg intraperitoneal for 3 days) or sham (0.6 ml saline IP for, 3 days) treatments. We measured the effects of IBU vs. sham in normoxic animals too. Expression of mRNA for IL‐1β, IL‐6, and TNFα measured with RT‐PCR in dorsal brainstem biopsies tended to be lower with IBU compared to sham: IBU = 68, 67 and 85% of sham levels for IL‐1β, IL‐6, and TNFα, respectively. However, ventilatory and metabolic responses to hypoxia measured with barometric pressure plethysmography and stop‐flow respirometry were not significantly different between IBU and sham in CSH or normoxic rats (p>;0.1). Hence, inflammatory cytokines in the CNS and carotid bodies appear necessary for the induction but not the maintenance of an increased hypoxic ventilatory response with CSH. Supported by NIH RO1 HL‐081823.

Effect ofWithania somniferaExtracts on Some Selective Biochemical, Hematological, and Immunological Parameters in Guinea Pigs Experimental Infected withE. coli

Fifty 1-2-month-old Guinea pigs were divided into 5 equal groups, 10 each. Control (Gp1) did receive neither viable bacteria nor treatment. Each animal from the other groups (Gp2–5) was challenged with (1-2 × 108) viable E. coli in 200 μL normal saline (0.9%) through IP route. GP2 infected group was treated with 200 μL saline IP and kept as positive control group. Gp3-4 are infected and treated with Withania somnifera (ethanol root extract) with doses 50 and 100 mg/kg. BW, respectively. Gp5 infected treated group was treated with cefoperazone antibiotic at dose 35 mg/Kg BW. The treatment by drug or the extracted medicinal plant was started 72 h after infection for 7 successive days. Serum and whole blood sample were collected from all groups 14 days after treatment to evaluate some hematological and biochemical changes as well as immunomodulatory cytokine tumor necrosis factor-alpha (TNF-α). Oral treatment of the plant extract caused significant benefit results in infected Guinea pig appeared in the correction of some hematological and biochemical parameters also try to suppressed inflammatory cytokine response represent in TNF-α. It could be concluded that W. somnifera extract has potent antibacterial activity, and this appears in the correction with hematological, biochemical, and immunological results.

Abstract TP263: The Effect Of Moderate And Excessive Alcohol Exposure On Male Experimental Stroke Outcomes In Mice Is Androgen-Independent

Heavy alcohol use is a major stroke risk factor while moderate alcohol use may offer protective benefits. We have shown that moderate ethanol (EtOH) exposure reduces while excessive EtOH exposure increases cortical infarct volume after experimental stroke in male and female mice. The effect of either moderate or excessive EtOH exposure levels in cortex is lost in ovariectomized mice, indicating female sex steroids may be involved in the female response to EtOH exposure and experimental stroke. However, it is unknown if male sex steroids (androgens) might alter the effect of varying EtOH exposure levels on male stroke outcomes. We determined the role of androgens in the male response to EtOH exposure and experimental stroke. Male mice were castrated (CAST) 7-8 days before EtOH exposure and middle cerebral artery occlusion (MCAO) to reduce endogenous androgens. Intact and CAST males were given 0.25 g/kg (moderate dose, n=11) or 2.5 g/kg (excessive dose, n=11) EtOH IP 1 h before 45 min of MCAO. These doses mimic moderate (1-2 drinks daily) or excessive (>4 drinks daily) alcohol use in men based on blood EtOH concentrations. Control male (n=11) and CAST mice (n=10) were given saline (0.015 mL/g) IP 1 h before MCAO. Infarct volume (% contralateral structure) was determined at 72 h reperfusion by analysis of coronal brain slices stained with 2,3,5-triphenyltetrazolium chloride. In contrast to saline treated mice (EtOH vs. saline), acute moderate EtOH exposure reduced (p<0.05) cortical infarct volume in intact (41+3% vs. 54+1%) and CAST (23+5% vs. 34+5%) males. Following acute excessive EtOH exposure, cortical infarct volume was increased (p<0.05) in male (62+2% vs. 54+1%) and CAST (47+4% vs. 34+5%) mice compared to saline treated mice. Clinically, our findings suggest that moderate alcohol use may positively while excessive alcohol use may negatively alter stroke outcomes in men regardless of androgen status. Future studies will need to address the role of androgens in aging male brain as gradual androgen loss during aging (andropause) may not have similar effects on the male brain’s response to EtOH exposure and experimental stroke as an abrupt loss of androgens in CAST mice.

Previous Burn Injury Predisposes Mice to Lipopolysaccharide-Induced Changes in Glucose Metabolism

In mice, it has been demonstrated that at 7 days after burn injury, injection of lipopolysaccharide (LPS) is more lethal than the same dose at 1 day after injury. In the present study, we examined the effect of LPS injection to mice burned 7 days previously on glucose metabolism ([(18)F] 2-fluoro-2-deoxy-D-glucose [(18)FDG] uptake) in vivo. CD-1 male mice (25-28 g, Charles River Breeding Laboratories, Wilmington, MA) were anesthetized, backs shaven, and subjected to dorsal full thickness burn on 25% TBSA. Sham-treated animals were used as controls. Six days after burn injury, all mice were fasted overnight. One half of the burned and sham controls were subsequently injected IP with LPS (10 mg/kg; Escherichia coli). The remaining animals were injected with saline IP. Two hours later, all mice were injected IV with 50 μCi of (18)F FDG. One hour later, the animals were euthanized, and biodistribution was measured. Tissues were weighed, and radioactivity was measured with a well-type γ counter. Results were expressed as %dose/g tissue, mean ± SEM. The combination of burn 7 days previously and LPS significantly increased mortality compared to animals with burn alone, LPS alone, or sham controls. Burn injury 7 days previously caused a significant decrease in (18)FDG uptake by the brain compared to sham controls. The combination of LPS and burn injury 7 days previously produced a significant increase in (18)FDG uptake by brown adipose tissue and heart compared with either treatment separately. LPS produced a significant increase in (18)FDG uptake by lung, spleen, and gastrointestinal tract of the sham animals, changes that were different in mice burned 7 days previously and injected with LPS. The present results suggest that burn injury 7 days previously predisposes mice to alterations in (18)FDG uptake produced by LPS. These changes may relate, in part, to the increased lethality of LPS injection in previously burned mice.

Microstructure of rat behavioral response to anxiety in hole-board

Aim of present research was to assess, by means of descriptive and multivariate approaches, in Wistar rats tested in hole-board apparatus, whether a microstructure of hole-exploratory behavior may be identified in the behavioral response to anxiety. Two groups of 15 subjects were tested: one received 1 ml of saline IP and one was IP injected with diazepam at 0.5 mg/kg dissolved in 1 ml of saline. Following saline, six not yet reported hole-exploratory components were identified: active-dip, static-dip, rapid-dip, circular-sniff, point-sniff and central-sniff. Diazepam provoked significant changes of their per cent distribution. Results of multivariate analyses revealed a complex microstructure of hole-exploratory elements characterized by behavioral flows between sniffing and dipping patterns. Diazepam significantly modified several patternings, mainly those involving point-sniff and static-dip. Hence, the hypothesis of an important role of such behavioral elements and of their patternings, in anxiety-related hole-exploratory behavior, can be proposed. It is suggested that modifications of point-sniff and static-dip patternings could evidence even subtle changes of animal anxiety condition.

Effects of different doses of ulinastatin on severe thermal injury-induced myocardial damage in rats

Objective To investigate the effects of different doses of ulinastatin on severe thermal injuryinduced myocardial damage in rats. Methods One hundred and eighty female SD rats weighing 180-220 g were usedin this study. Thirty percent of the total body surface (TBS) was shaved chemically with 20% sodium sulphate and then exposed to 92 ℃ water for 18 s. The animals with third degree thermal injury involving 30% of the TBS were randomly divided into 3 groups (n = 60 each): group Ⅰ thermal injury (group TI); group Ⅱ and Ⅲ received intraperitoneal ulinastatin 40 000 and 80 000 U/kg respectively immediately after thermal injury (group U1 , U2). The TI group received equal volume of normal saline IP instead of ulinastatin. Blood samples were taken from abdominal aorta before (baseline) and at 1, 3, 6, 12 h after thermal injury for determination of serum concentrations of cTnI, IL-1β, IL-6, IL-10 and TNF-α (10 samples at each time points). Ten animals were sacrificed at each time point after blood sampling. The myocardial specimens were obtained for microscopic examination and measurement of MDA content and SOD activity. Results Compared with group TI, the serum concentrations of cTnI, IL-1β, IL-6 and TNF-α and MDA content in myocardium were significantly decreased and the myocardial SOD activity was significantly increased in group U1 , while in group U2 the senum concentrations of cTnI, IL-Iβ, IL-6 and TNF-α and myocardial MDA content were significantly increased and the myocardial SOD activity was significantly decreased. There was no siginificant difference in the serum concentrations of IL-10 among the three groups. Microscopic examination showed that myocardial damage was accentuated in group U2 as compared with group U1. Conclusion Ulinastatin 40 000 U/kg can ameliorate severe thermal injury-induced myocardial injury through inhibition of inflammatory response and lipid peroxidation response, whereas ulinastatin 80 000 U/kg accentuates myocardial damage. Key words: Trypsin inhibitors; Burns; Myocardium

Longitudinal microPET imaging of brain glucose metabolism in rat lithium–pilocarpine model of epilepsy

The lithium–pilocarpine model of epilepsy in rat has been used extensively to investigate basic mechanisms of epilepsy and mimics human temporal lobe epilepsy. Our aim was to investigate longitudinal alterations in metabolism after lithium–pilocarpine induced status epilepticus (SE) using [18F]FDG microPET. Twenty-eight Wistar rats received lithium chloride followed by pilocarpine (n=19) or saline (n=9) IP. Continuous video-EEG was used to monitor SE and occurrence of spontaneous seizures (SS). FDG microPET imaging was performed at baseline, on day 3 after drug administration (D3), and at the end of the monitoring period (CR). MicroPET images were spatially normalized to Paxinos space and parametric standardized uptake value (SUV)-images were generated. Metabolism was compared between groups of animals and between different time points. Eighteen animals developed SE, 11 had died by D3. SS were recorded in 3 of 7 surviving SE animals. On D3, metabolism was reduced in SE group compared to controls throughout the brain (−49±27%), except for the cerebellum: mostly in hippocampus, entorhinal cortex and thalamus bilaterally. Metabolism tended to be different between SS and no SS animals on D3 in striatum and hippocampus. In CR condition, relative metabolism was significantly different in SE group compared to controls in cerebellum and brainstem bilaterally and left striatum and entorhinal cortex. There were no significant differences between SS and no SS animals in CR condition. Pilocarpine-induced SE causes a severe, but transient reduction in overall metabolism on D3 in rat brain. Metabolic differences on D3 between SS and no SS animals need further study to investigate potential use as an early marker of epileptogenesis.