Male Swiss-Webster Mice Research Articles

Pharmacokinetics of Synthetic Cathinones Found in Bath Salts in Mouse Brain and Plasma Using High-Pressure Liquid Chromatography-Tandem Mass Spectrometry.

Approximately 10 years ago, "bath salts" became popular as legal alternatives to the psychostimulants cocaine and the amphetamines. These products contained synthetic cathinones, including 3,4-methylenedioxypyrovalerone (MDPV), 4-methylmethcathinone (mephedrone), and 3,4-methylenedioxymethcathinone (methylone). Most preclinical investigations have only assessed the effects of these synthetic cathinones independently; however, case reports and Drug Enforcement Administration (DEA) studies indicate that bath salts contain mixtures of these substances. In this study, we examine the pharmacokinetic interactions of the drug combination. We hypothesized that combined exposure to MDPV, mephedrone, and methylone would result in increased drug concentrations and enhanced total drug concentrations when compared to individual administration. Adolescent male Swiss-Webster mice were injected intraperitoneally with either 10 mg/kg MDPV, 10 mg/kg mephedrone, 10 mg/kg methylone, or 10 mg/kg combined MDPV, mephedrone, and methylone. Following injection, brains and plasma were collected at 1, 10, 15, 30, 60, and 120 min. Drugs were extracted via solid-phase extraction, and concentrations were determined using a previously published high-pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. All drugs crossed the blood-brain barrier quickly. For methylone, the maximal concentration (Cmax) and the total drug exposure [as represented by the area under the concentration-time curve (AUC)] were significantly higher when combined with mephedrone and MDPV in both matrices (2.89-fold increase for both Cmax and AUC with combined treatment). For mephedrone, the Cmax was unchanged, but the AUC in brain was increased when in combination by approximately 34%. Interestingly, for MDPV, the Cmax was unchanged, yet the AUC was higher when MDPV was administered individually (there was a 62% decrease in AUC with combined treatment). The pharmacokinetics of methylone, mepedrone, and MDPV are altered when the drugs are used in combination. These data provide insight into the consequences of co-exposure to synthetic cathinones in popular bath salt products.

Fentanyl-related substances elicit antinociception and hyperlocomotion in mice via opioid receptors

Synthetic opioids have been implicated as the single greatest contributor to rising drug-related fatalities in recent years. This study evaluated mu-opioid receptor (MOR) mediated effects of seven fentanyl-related substances that have emerged in the recreational drug marketplace, and for which there are no existing or only limited in vivo data. Adult male Swiss Webster mice were administered fentanyl-related substances and their effects on nociception and locomotion as compared to MOR agonist standards were observed. In locomotor activity tests, morphine (100, 180 mg/kg), fentanyl (1, 10 mg/kg), beta-methylfentanyl (10 mg/kg), para-methoxyfentanyl (10 mg/kg), fentanyl carbamate (100 mg/kg), and 3-furanylfentanyl (10 mg/kg), elicited significant (p ≤ 0.05) dose-dependent increases in locomotion. However, para-methylfentanyl and beta′-phenylfentanyl did not produce significant effects on locomotion at doses up to 100 mg/kg and phenylfentanyl (100 mg/kg) significantly decreased locomotion. In warm-water tail-withdrawal tests, all substances produced significant dose-dependent increases in antinociception with increasing ED50 values (95% CI) of fentanyl [0.08 mg/kg (0.04–0.16)] > para-methoxyfentanyl [0.43 mg/kg (0.23–0.77)] > 3-furanylfentanyl [0.51 mg/kg (0.36–0.74)] > beta-methylfentanyl [0.74 mg/kg (0.64–0.85)] > para-methylfentanyl [1.92 mg/kg (1.48–2.45)] > fentanyl carbamate [5.59 mg/kg (4.11–7.54)] > morphine [7.82 mg/kg (5.42–11.0)] > beta′-phenylfentanyl [19.4 mg/kg (11.0–34.4)] > phenylfentanyl [55.2 mg/kg (33.5–93.0)]. Naltrexone (1 mg/kg) increased ED50 values several fold with decreasing magnitudes of para-methylfentanyl (63.1×) > para-methoxyfentanyl (22.5×) > beta′-phenylfentanyl (21.0×) > 3-furanylfentanyl (20.6×) > beta-methylfentanyl (19.2×) > phenylfentanyl (5.23×) > fentanyl (3.95×) > fentanyl carbamate (2.21×) > morphine (1.48×). These findings expand upon in vivo results from previous studies and establish that the effects of these fentanyl related-related substances are at least in part mediated by the MOR.

Ascorbic acid inhibits the acquisition and expression of morphine-induced conditioned place preference and sensitization in male Swiss-Webster mice

Ascorbic acid inhibits the acquisition and expression of morphine-induced conditioned place preference and sensitization in male Swiss-Webster mice

Effect of Curcuma longa Crude Extract, Curcumin and Nano Curcumin-based Gel Topical Administration on Excised Skin Wound Healing in Swiss-Webster Mice

Patients suffering from skin wounds often have their life heavily affected economically and socially. In general, wound healing consists of four different sequentially and overlapping phases. Interrupting those phases results in extending the healing time. Turmeric (Curcuma longa) is widely used by Indonesians to treat wounds, with curcumin as an active substance, which is proven to have various effects on improving wound healing. Therefore, this study aims to determine how the topical application of turmeric gel on affects the acceleration of wound closure. Analytical laboratory experiments were conducted on 24 male Swiss-Webster mice. Subjects were divided into four groups according to where the excision wounds were made on the back, and observed on days 0, 3, 6, 9, 12, and 15 post wound induction. The wound healing acceleration was observed after 3 (p = 0.0003), 6 (p = 0.0085), and 9 (p = 0.0023) days in C. longa-treated mice compared to the control group. However, there was no significant difference found on days 12 and 15 after wounding. In addition, the area under the curve analysis shows significant difference between the C. longa-treated group compared to the control, but not with the group treated with neither curcumin nor nano curcumin gel. In conclusion, topical application of 3% C. longa extract gel improves excised skin healing.

Immunostimulatory Activity Test of Extract and Isolation of the Secondary Metabolites of Kabau (Archidendron Bubalinum) Fruit Peel

Kabau fruit peel has higher antioxidant activity. Antioxidants can changes immunological and oxidative stress parameters and enhance the immune system. This study aimed to determine the immunostimulatory activity of 96 % and 50 % ethanol extracts of kabau fruit peel and to isolate the major secondary metabolites. The immunostimulatory activity was tested using the carbon clearance method. Eighteen male Swiss Webster mice were divided into groups, given each ethanol extract at a respective dose of 100 and 200 mg/kg BW, 0.5 % Na-CMC as the negative control, and 19.5 mg/kg BW Stimuno as the positive control. Each treatment was given for seven days. The active extract was further fractionated and isolated using chromatography, followed by identification and characterization of the isolate using TLC-Scanner and NMR. The 96 % ethanol extract had immunostimulatory activity, with a phagocytic index of 1.17. Isolation of the ethanol extract revealed Quercitrin, a flavonoid, with Rf value of 0.55. Results of this study indicate that kabau fruit peel extract has immunostimulatory activity with flavonoid as the active ingredient.

Respiratory Depressant Effects of Fentanyl Analogs

Aim Opioid-related fatalities involving synthetic narcotics have reached unprecedented levels. This study evaluated the respiratory depressant effects of seven fentanyl analogs that have either emerged in the recreational drug marketplace or been identified in toxicological analyses following fatal or non-fatal intoxications and for which their effects on ventilation had not been previously characterized. Methods Adult male Swiss Webster mice (N = 8/group) were administered fentanyl analogs (isobutyrylfentanyl, crotonylfentanyl, para-methoxyfentanyl, para-methoxybutyrylfentanyl, 3-furanylfentanyl, thiophenefentanyl, benzodioxolefentanyl) and their effects on respiratory rate, tidal volume, and minute volume as compared to mu-opioid-receptor (MOR) agonist standards (fentanyl, morphine, and buprenorphine) were measured using whole body plethysmography (WBP). Results All compounds elicited significant (p ≤ 0.05) hypoventilation relative to vehicle at least at one dose tested: morphine (1, 3.2, 10, 32 mg/kg), buprenorphine, (0.032, 0.1, 0.32, 1, 3.2 mg/kg), fentanyl (0.01, 0.032, 0.1, 1, 32 mg/kg), isobutyrylfentanyl (0.1, 0.32, 1, 3.2, 10 mg/kg), crotonylfentanyl (0.1, 0.32, 1, 3.2, 10 mg/kg), para-methoxyfentanyl (0.1, 0.32, 1, 3.2, 10 mg/kg), para-methoxybutyrylfentanyl (0.32, 1, 3.2, 10 mg/kg), 3-furanylfentanyl (0.1, 0.32, 1, 3.2, 10 mg/kg), thiophenefentanyl (1, 3.2, 10, 32, 100 mg/kg), and benzodioxolefentanyl (3.2, 10, 32, 100 mg/kg). The ED50 values for hypoventilation showed a rank order of potency as follows: fentanyl (ED50 = 0.96 mg/kg) > 3-furanylfentanyl (ED50 = 2.60 mg/kg) > crotonylfentanyl (ED50 = 2.72 mg/kg) > para-methoxyfentanyl (ED50 = 3.31 mg/kg) > buprenorphine (ED50 = 10.8 mg/kg) > isobutyrylfentanyl (ED50 = 13.5 mg/kg) > para-methoxybutyrylfentanyl (ED50 = 16.1 mg/kg) > thiophenefentanyl (ED50 = 18.0 mg/kg) > morphine (ED50 = 55.3 mg/kg) > benzodioxolefentanyl (ED50 = 10168 mg/kg). A naloxone pretreatment (10 mg/kg) attenuated the hypoventilatory effects of all drugs. Conclusions These results establish that the respiratory depressant effects of these fentanyl analogs are at least in part mediated by the MOR.

THE POTENTIAL OF NGOKILO LEAVES EXTRACT (STACHYTARPHETA MUTABILIS. VAHL) AS A LOWERING OF BLOOD GLUCOSE LEVELS OF WHITE MICE (RATTUS NORVEGICUS)

Diabetes is a disease, in which the body cannot produce enough insulin so that there is excess sugar in the blood which becomes toxic to the body. Ngokilo plants are believed to be able to lower blood sugar levels because of the antioxidants and polyphenols contained therein.The objective of this study was to determine the potential of the ethanol extract of Ngokilo leaves (Stachytarpheta mutabilis, Vahl) as a lowering blood glucose levels in white mice(Rattus norvegicus). The tests were carried out by an experimental method using Swiss Webster male mice (Rattus norvegicus) aged 2-3 months with a body west of 30-40 grams as test animals. This study used four dosage ratios of ngokilo leaf extract, namely 1: 20: 40: 60 and control used aquadest. The results showed that the ethanol extract of Ngokilo leaves had an F value of 27.033> F Table 2.60. It indicates that the independent variable (the difference in the concentration of ethanol extract of Ngokilo leaves) was proven to have a significant effect on the dependent variable (blood sugar levels).

MA-[D-Leu-4)-OB3, a Small Molecule Synthetic Peptide Leptin Mimetic, Improves Serum Lipid Profiles in Mouse Models of Type 1 and Type 2 Diabetes Mellitus

Diabetic dyslipidemia, a common pattern of lipid abnormalities associated with diabetes, is characterized by hyperglycemia, elevated serum levels of triglycerides and low-density lipoprotein (LDL)-cholesterol, and reduced serum levels of high-density lipoprotein (HDL)-cholesterol. These lipid abnormalities make diabetes an independent risk-factor for the development of cardiovascular disease. Two mouse models of diabetes were used in the present study: 1) streptozotocin (STZ)-induced insulin-deficient male Swiss Webster mice (a model of T1DM), and 2) male diet-induced obese (DIO) mice rendered insulin-resistant by maintenance on a high-fat diet for 30 weeks (a model of T2DM). Both models were then treated with MA-[D-Leu-4]-OB3 by oral gavage for 14 or 17 days. At the end of the test period, the mice were exsanguinated by cardiac puncture; serum was prepared from the whole blood and analyzed by ELISA for total-, HDL-, LDL/VLDL-cholesterol, and triglyceride content. In mouse models of both T1DM and T2DM, MA-[D-Leu-4]-OB3 significantly (P < 0.05) reduced serum levels of triglycerides and LDL/VLDL-cholesterol, and elevated serum HDL-cholesterol to levels approaching or equivalent to those seen in normal non-diabetic mice of the same age and sex. Our results suggest that in addition to its effects on energy balance, glycemic control, and cognitive function, MA-[D-Leu-4]-OB3, by modulating serum lipid levels, may also be beneficial in reducing the risk of adverse cardiovascular events associated with both T1DM and T2DM.

PENGARUH AIR REBUSAN DAUN HARENDONG BULU (Clidemia hirta) TERHADAP KADAR GULA DARAH MENCIT (Mus musculus)

This study aims to determine the effect of the water boiled leaves of Clidemia hirta on blood sugar levels in Swiss Webster mice (Mus musculus). This research using a completely randomized design research design (CRD) with 5 treatment groups namely; Aquades control group (P0), 3 g sucrose treatment (P1), 3 g sucrose treatment + boiled bristle leaves boiled water at a dose of 0.13 ml/KgBB (P2), 3 g sucrose treatment + boiled bristle leaves boiled water at a dose of 0.26 ml/KgBB (P3), 3 g treatment of sucrose + boiled bristle leaves boiled water at a dose of 0.39 ml/KgBB (P4). Before administration of the extract, mice in the treatment groups P1, P2, P3 &amp; P4 were induced using 3 g of sucrose for 5 days. Then after 5 days for groups P2, P3, and P4 followed by giving boiled water leaves for 7 days. The analysis result of One Way Anova Test shows that Fcount &gt; Ftable (18.86 &gt; 2.51). This shows the boiled water affects the blood sugar levels of male Swiss Webster mice. The concentration of extract at P4 (0.39 ml/KgBB) was more effective in reducing blood sugar levels.

PENGARUH AIR REBUSAN DAUN HARENDONG BULU (Clidemia hirta) TERHADAP KADAR KOLESTROL MENCIT (Mus musculus)

This study aims to determine the effect of giving boiled water leaves Clidemia hirta on the levels of male webster swiss mice cholesterol. This type of research is an experimental study using the Completely Randomized Design (CRD) method which consists of 5 treatments with 5 repetitions. The experimental animals in this study were male mice which were divided into 5 groups namely P0 (aquadest), P1 (MDLT + PTU + boiled water 0.13 ml/bb), P2 (MDLT + PTU + boiled water 0.26 ml/bb), P3 (MDLT + PTU + boiled water 0.39 ml/bb), and P4 (Simvastatin 10 mg). The treatment was carried out using the gavage method using a gastric sonde device, giving boiled bristle leaves boiled water in the morning for 7 consecutive days carried out in the morning. Retrieval of blood cholesterol levels data by cutting the tail of mice and the next drop blood on the cholesterol strip Autocheck tool. Based on the results of the Kruskal Wallis test calculation states that Hcount &lt;Htable (4.626 &lt;9.49), it can be concluded that the provision of boiled water leaves of Clidemia hirta does no effect significant in reducing levels of male Swiss Webster mice (Mus musculus) cholesterol levels.

Noise Exposure Decreases the Number of Spermatozoa in Male Mice (Mus musculus L.) Even With Grape Seed Extract (Vitis vinifera L.) Administration

Excessive continuous noises exposure changes the male hormone system which leads to formation of oxidative stress and results in disrupt of semen quality. This condition can be reduced by the use of antioxidants. Grape seed is one of the antioxidants that contains phenol components that have Resveratrol compounds. This research aimed to observe the effect of noise exposure on the number of spermatozoa of male mice given grape seed extract. The method of the research was a post-test only control group design with research subjects of 30 male Mus musculus L. (Swiss Webster) mice divided into 5 groups: group A (treatment control) was not given noise exposure and grape seed extract, group B (negative control) was not given noise exposure and was given grape seed extract, groups C, D, E (treatment group) were given noise exposure with sequential intensities of 65 dB, 85 dB, 105 dB. The treatment was given for 33 days. The results showed that noise exposure with different intensities of 65 dB, 85 dB, 105 dB reduced the spermatozoa count of male Swiss Webster mice even with the administration of grape seed extract. One Way Anova test was used to analyze the data with p-value of 0.001. Conclusion: There were differences in spermatozoa count between the control group and the treatment group. Provision of noise exposure with a value above the threshold limit reduces the number of male Swiss Webster mice spermatozoa given with grape seed extract.&#x0D; Keywords: Number of Spermatozoa, Grape Seed Extract, Noise exposure&#x0D;

Mitochondrial-Protective Effects of R-Phenibut after Experimental Traumatic Brain Injury.

Altered neuronal Ca2+ homeostasis and mitochondrial dysfunction play a central role in the pathogenesis of traumatic brain injury (TBI). R-Phenibut ((3R)-phenyl-4-aminobutyric acid) is an antagonist of the α2δ subunit of voltage-dependent calcium channels (VDCC) and an agonist of gamma-aminobutyric acid B (GABA-B) receptors. The aim of this study was to evaluate the potential therapeutic effects of R-phenibut following the lateral fluid percussion injury (latFPI) model of TBI in mice and the impact of R- and S-phenibut on mitochondrial functionality in vitro. By determining the bioavailability of R-phenibut in the mouse brain tissue and plasma, we found that R-phenibut (50 mg/kg) reached the brain tissue 15 min after intraperitoneal (i.p.) and peroral (p.o.) injections. The maximal concentration of R-phenibut in the brain tissues was 0.6 μg/g and 0.2 μg/g tissue after i.p. and p.o. administration, respectively. Male Swiss-Webster mice received i.p. injections of R-phenibut at doses of 10 or 50 mg/kg 2 h after TBI and then once daily for 7 days. R-Phenibut treatment at the dose of 50 mg/kg significantly ameliorated functional deficits after TBI on postinjury days 1, 4, and 7. Seven days after TBI, the number of Nissl-stained dark neurons (N-DNs) and interleukin-1beta (IL-1β) expression in the cerebral neocortex in the area of cortical impact were reduced. Moreover, the addition of R- and S-phenibut at a concentration of 0.5 μg/ml inhibited calcium-induced mitochondrial swelling in the brain homogenate and prevented anoxia-reoxygenation-induced increases in mitochondrial H2O2 production and the H2O2/O ratio. Taken together, these results suggest that R-phenibut could serve as a neuroprotective agent and promising drug candidate for treating TBI.

MA-[D-Leu-4]-OB3, a small molecule synthetic peptide leptin mimetic, improves episodic memory, and reduces serum levels of tumor necrosis factor-alpha and neurodegeneration in mouse models of Type 1 and Type 2 Diabetes Mellitus

BackgroundExtracellular beta-amyloid (Aβ), intra-neuronal hyper-phosphorylated tau protein, and chronic inflammation are neuropathological hallmarks of Alzheimer's Disease (AD). A link between AD, insulin dysfunction, and tumor necrosis factor-alpha (TNF-α) in promoting both tau and Aβ pathologies in vivo has been proposed. MethodsMA-[D-Leu-4]-OB3 was given, with or without insulin, to streptozotocin (STZ)-treated male Swiss Webster mice, and to male diet-induced obese (DIO) mice. Brains were excised, and coronal sections were imaged with fluoro jade-C (FJC), thioflavin-S, or hematoxylin and eosin (H&E). Serum TNF-α and IGF-1 were measured by ELISA. Histopathological changes in the cerebral cortex (CC) and hippocampus (HC) were correlated with changes in glycemic regulation, episodic memory, and serum levels of TNF-α and IGF-1. ResultsIn STZ-treated mice, blood glucose and serum TNF-α and IGF-1 were reduced by insulin alone, and normalized when MA-[D-Leu-4]-OB3 was given in combination with insulin. Improvement in episodic memory was inversely correlated with the number of FJC-positive cells in the CC and HC and serum TNF-α and IGF-1. FJC, thioflavin-S and H&E staining indicated no Aβ deposition. Similar results were observed in DIO mice treated with MA-[D-Leu-4]-OB3. ConclusionsThe mechanism by which MA-[D-Leu-4]-OB3 improves episodic memory in mouse models of TIDM and T2DM appears to be related to improved insulin sensitivity and reduced TNF-α-induced neurodegeneration. General significanceMA-[D-Leu-4]-OB3 may have application to human pre-clinical and clinical AD and AD-like dementia by interrupting the cascade of insulin resistance, neuro-inflammation, and neurodegeneration, that characterizes these diseases.

Activation of PPARγ reduces N-acetyl-cysteine -induced hypercorticoidism by down-regulating MC2R expression into adrenal glands

We previously demonstrated that oral supplementation with antioxidants induced hyperactivity of hypothalamus-pituitary-adrenal (HPA) axis, attested by hypercorticoidism, through an up-regulation of adrenocorticotrophic hormone (ACTH) receptors (MC2R) in adrenal. This study analyzed the role of peroxisome proliferator-activated receptor (PPAR)-γ on HPA axis hyperactivity induced by N-acetyl-cysteine (NAC). Male Swiss-Webster mice were orally treated with NAC for 1, 3, 5, 10, 15, or 18 consecutive days. The PPAR-γ agonist rosiglitazone and/or antagonist GW9662 were daily-injected i.p. for 5 consecutive days, starting concomitantly with NAC treatment. Rosiglitazone treatment inhibited NAC-induced adrenal hypertrophy and hypercorticoidism. Rosiglitazone also significantly reversed the NAC-induced increase in the MC2R expression in adrenal, but not steroidogenic acute regulatory protein (StAR). NAC treatment reduces the expression of PPARγ in the adrenals, but rosiglitazone did not restore the expression of this cytoprotective gene. In addition, GW9662 blocked the ability of rosiglitazone to decrease plasma corticosterone levels in NAC-treated mice. In conclusion, our findings showed that antioxidant supplementation induced a state of hypercorticoidism through down-regulation of PPARγ expression in the adrenals, in a mechanism probably related to a down-regulation of ACTH receptor expression.

The Difference of Effectivity Between Nutmeg Seed Extract (Myristica Fragrans Houtt) and Diazepam Based on Sleep Induction Time on Swiss Webster Mice

According to the National Sleep Foundation in America, more than one third (36%) of young adults aged 18-29 years reported having difficulty getting up early (compared to 20% at 30-64 years old and 9% over 65 years). The National Sleep Foundation also states that in Indonesia the prevalence of insomnia sufferers reaches 70% at least once a week and 30 million people have difficulty sleeping every night. This study is an experimental study with a post-test only control group design. The samples used were 27 male Swiss webster mice which were divided into three groups. The treatment group was given nutmeg seed extract at a dose of 7.5 mg / kgBW; the positive control group was given diazepam, and the negative control group was given aquabidest. The results showed the difference in the average value of sleep induction time in the treatment group was 2.97 minutes, positive control group 5.06 minutes, and negative control group 8.43 minutes with p = 0.00. Conclusion: There is a significant difference between the effectiveness of nutmeg seed extract 7.5 mg / kgBW with diazepam based on the time of sleep induction in Swiss webster male mice.

The test anti-inflammatory activity of infusing Bryophyllum pinnatum (Kalanchoe pinnata) leaves (Kalanchoe pinnata) on edema in mice leg thigh male swiss webster

Research to know the effect of anti-inflammatory of infuse bryophyllum pinnatum (Kalanchoe pinnata) against edema on leg thigh Swiss Webster male mice and to see relationships increased concentrations with increased anti-inflammatory effect. This research was conducted in the Science Laboratory, Faculty of Fisheries, Agriculture, and Biology of the Bangka Belitung University. The method: The animals test that used were 25 Swiss Webster male mice were divided into 5 groups, namely group I: negative control (aquadest), group II: positive control (aspirin), group III, IV, V given infuse bryophyllum pinnatum (Kalanchoe pinnata) with concentrations 20%, 40%, and 80%. This research uses a method by injecting acetic acid in mice thigh. The measured data is a murine thigh volume for 3 hours using a caliper. Data analysis using the Kruskal-Wallis, the results showed that there were no significant differences as a value greater than 0.05 is 0.533. This suggests that increasing the concentration did not give a considerable influence on the volume of edema thigh male mice Swiss Webster. From the Mann Whitney test between the concentration of 20%, 40%, 80%, and negative controls showed that the difference in significance because of the value of significantly less than 0,05. This shows that there is a potential infusion of leaves bryophyllum pinnatum (Kalanchoe pinnata) towards healing leg edema in the thigh swiss webster male mice.

Mesenchymal Stromal Cells Protect the Blood-Brain Barrier, Reduce Astrogliosis, and Prevent Cognitive and Behavioral Alterations in Surviving Septic Mice.

Survivors of sepsis are frequently left with significant cognitive and behavioral impairments. These complications derive from nonresolving inflammation that persists following hospital discharge. To date, no study has investigated the effects of mesenchymal stromal cell therapy on the blood-brain barrier, astrocyte activation, neuroinflammation, and cognitive and behavioral alterations in experimental sepsis. Prospective, randomized, controlled experimental study. Government-affiliated research laboratory. Male Swiss Webster mice (n = 309). Sepsis was induced by cecal ligation and puncture; sham-operated animals were used as control. All animals received volume resuscitation (1 mL saline/mouse subcutaneously) and antibiotics (meropenem 10 mg/kg intraperitoneally at 6, 24, and 48 hours). Six hours after surgery, mice were treated with mesenchymal stromal cells IV (1 × 10 cells in 0.05 mL of saline/mouse) or saline (0.05 mL IV). At day 1, clinical score and plasma levels of inflammatory mediators were increased in cecal ligation and puncture mice. Mesenchymal stromal cells did not alter clinical score or survival rate, but reduced levels of systemic interleukin-1β, interleukin-6, and monocyte chemoattractant protein-1. At day 15, survivor mice completed a battery of cognitive and behavioral tasks. Cecal ligation and puncture mice exhibited spatial and aversive memory deficits and anxiety-like behavior. These effects may be related to increased blood-brain barrier permeability, with altered tight-junction messenger RNA expression, increased brain levels of inflammatory mediators, and astrogliosis (induced at day 3). Mesenchymal stromal cells mitigated these cognitive and behavioral alterations, as well as reduced blood-brain barrier dysfunction, astrocyte activation, and interleukin-1β, interleukin-6, tumor necrosis factor-α, and interleukin-10 levels in vivo. In cultured primary astrocytes stimulated with lipopolysaccharide, conditioned media from mesenchymal stromal cells reduced astrogliosis, interleukin-1β, and monocyte chemoattractant protein-1, suggesting a paracrine mechanism of action. In mice who survived experimental sepsis, mesenchymal stromal cell therapy protected blood-brain barrier integrity, reduced astrogliosis and neuroinflammation, as well as improved cognition and behavior.

Toxicity and occupational exposure assessment for hydroprocessed esters and fatty acids (HEFA) alternative jet fuels

ABSTRACTThe U.S. Air Force (USAF) has pursued development of alternative fuels to augment or replace petroleum-based jet fuels. Hydroprocessed esters and fatty acids (HEFA) renewable jet fuel is certified for use in commercial and USAF aircraft. HEFA feedstocks include camelina seed oil (Camelina sativa, HEFA-C); rendered animal fat (tallow, HEFA-T); and mixed fats and oils (HEFA-F). The aim of this study was to examine potential toxic effects associated with HEFA fuels exposures. All 3 HEFA fuels were less dermally irritating to rabbits than petroleum-derived JP-8 currently in use. Inhalation studies using male and female Fischer-344 rats included acute (1 day, with and without an 11-day recovery), 5-, 10- or 90-day durations. Rats were exposed to 0, 200, 700 or 2000 mg/m3 HEFA-F (6 hr/day, 5 days/week). Acute, 5 – and 10-day responses included minor urinalysis effects. Kidney weight increases might be attributed to male rat specific hyaline droplet formation. Nasal cavity changes included olfactory epithelial degeneration at 2000 mg/m3. Alveolar inflammation was observed at ≥700 mg/m3. For the 90-day study using HEFA-C, no significant neurobehavioral effects were detected. Minimal histopathological effects at 2000 mg/m3 included nasal epithelium goblet cell hyperplasia and olfactory epithelium degeneration. A concurrent micronucleus test was negative for evidence of genotoxicity. All HEFA fuels were negative for mutagenicity (Ames test). Sensory irritation (RD50) values were determined to be 9578 mg/m3 for HEFA-C and greater than 10,000 mg/m3 for HEFA-T and HEFA-F in male Swiss-Webster mice. Overall, HEFA jet fuel was less toxic than JP-8. Occupational exposure levels of 200 mg/m3 for vapor and 5 mg/m3 for aerosol are recommended for HEFA-based jet fuels.

Anxiogenesis induced by social defeat in male mice: Role of nitric oxide, NMDA, and CRF1 receptors in the medial prefrontal cortex and BNST

Nitric oxide (NO) release in the right medial prefrontal cortex (RmPFC) produces anxiogenesis. In the bed nucleus of the stria terminalis (BNST), a region that receives neuronal projections from the mPFC, NO provokes anxiety, an effect that is blocked by local injections of corticotrophin-releasing factor type 1 receptor (CRF1) or n-methyl-d-aspartate receptor (NMDAr) antagonist. Anxiety is also enhanced by social defeat stress, and chronic stress impairs and facilitates, respectively, PFC and BNST roles in modulating behavioral responses to aversive situations. This study investigated whether the (i) chronic social defeat stress (CSDS) increases NO signaling in the mPFC; and/or (ii) anxiogenic effects provoked by the intra-RmPFC injection of NOC-9 (an NO donor) or by CSDS are prevented by intra-BNST injections of AP-7 (0.05 nmol) or CP 376395 (3.0 nmol), respectively, NMDAr and CRF1 antagonists, in male Swiss-Webster mice exposed to the elevated plus-maze (EPM). Results showed that (a) CSDS increased anxiety (i.e., reduced open-arm exploration) and repeatedly activated nNOS-containing neurons, as measured by ΔFosB (a stable nonspecific marker of neural activity) + nNOS double-labeling, in the right (but not left) mPFC, (b) NOC-9 in the RmPFC also increased anxiety, and (c) both CSDS and NOC-9 effects were reversed by injections of AP-7 or CP 376395 into the BNST. These results suggest that NMDA and CRF1 receptors located in BNST play an important role in the modulation of anxiety provoked by NO in the RmPFC, as well as by chronic social defeat in mice.

Environmentally relevant exposures of male mice to carbendazim and thiram cause persistent genotoxicity in male mice.

Carbendazim and thiram are fungicides used in combination to prevent mold destruction of crops. Studies have demonstrated genotoxicity by these agents, but have not used concentrations below their water solubility limits in drinking water to test for persistence of genotoxicity due to chronic exposure. Ten 8-week old male Swiss-Webster mice were exposed to tap water, or nominal concentrations of 20μM carbendazim, 20μM thiram or 20μM of both fungicides for 90days (total of 40 mice). Five mice from tap water controls, carbendazim, thiram and combination-treated groups (20 mice total) had genotoxicity detected by comet assay of lymphocytes at the termination of the exposure period. The other 20 mice (4 treatment groups) were all switched to tap water and allowed a 45-day recovery period to check for persistence of DNA damage. The damage was compared with commercial control cells exposed to increasingly harsh treatment by etopside. Comet assay (mean % tail DNA + SE) of control mice (9.8+0.9) was similar to commercial control (CC0) cells (8.5+0.9). Carbendazim, thiram or the combination treatment caused similar mean % tail DNA with 33.0+2.9, 30.1+3.3 and 29.1+1.8, respectively, comparable with commercial cells slightly damaged by etopside (CC1 with 31.4+2.9) with no statistical change in water or food intake, body weight or liver or kidney weights. The key result was that a 45-day recovery period had no observable difference in the DNA damage as assessed by DNA % in comet tail with tap water controls and CCO control cells at 7.0+0.7 and 9.7+1.2 versus 27.5+1.9, 29.3+2.2 and 32.0+1.8, respectively, for carbendazim, thiram and combination treatments. It is of concern that the use of these agents in developing countries with little training or regulation results in water pollution that may cause significant persistent DNA damage in animal or human populations that may not be subject to repair.