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Great adaptability of brown adipose tissue mitochondria to extreme ambient temperatures in control and cold-acclimated gerbils as compared with mice.

1. The gerbil (Gerbillus campestris) is a desert rodent able to tolerate high (38 degrees C) and low (-20 degrees C) ambient temperatures, probably due to both its low resting metabolic rate in hot environment and its high peak metabolic rate in cold. 2. Measurement of mitochondrial state IV respiration and cytochrome-oxidase activity (COX) were made in interscapular brown adipose tissue (IBAT), liver and hind limb muscles of gerbils and mice of nearly equal body mass, acclimated for 4 weeks at cold ambient temperature (CA) or reared at thermoneutrality (TN). 3. The most striking difference between these two animal species appears to be in IBAT mitochondria: in TN animals, the level of state IV respiration and COX activity was lower in gerbils than in mice, but the cold acclimation-induced increase in these parameters was greater in gerbils than in mice. 4. Alternatively, in gerbils as in mice, cold acclimation induced a reduction in muscle mitochondrial COX activity. No important change due to cold acclimation was observed in liver mitochondria, either in gerbils or in mice. 5. As compared with mice, the lower state IV respiration in IBAT mitochondria from TN gerbils may explain their low RMR, whereas the higher COX activity of IBAT mitochondria from CA gerbils may explain their higher PMR. 6. As a result of this great adaptability of BAT mitochondria, the gerbil seemed to be able to live in a wide range of ambient temperatures in its natural habitat.

Acclimatization↗

Oral or parenteral administration of replication-deficient adenoviruses expressing the measles virus haemagglutinin and fusion proteins: protective immune responses in rodents.

The genes encoding the measles virus (MV) haemagglutinin (H) and fusion (F) proteins were placed under the control of the human cytomegalovirus immediate early promoter in a replication-deficient adenovirus vector. Immunofluorescence and radioimmune precipitation demonstrated the synthesis of each protein and biological activity was confirmed by the detection of haemadsorption and fusion activities in infected cells. Oral as well as parenteral administration of the H-expressing recombinant adenovirus elicited a significant protective response in mice challenged with MV. While the F-expressing adenovirus failed to protect mice, cotton rats immunized with either the H- or F-expressing recombinant showed reduced MV replication in the lungs. Antibodies elicited in mice following immunization with either recombinant had no in vitro neutralizing activity, suggesting a protective mechanism involving a cell-mediated immune response. This study demonstrates the feasibility of using oral administration of adenovirus recombinants to induce protective responses to heterologous proteins.

Adenoviridae↗

Nitric oxide-dependent modulation of sympathetic neural control of oxygenation in exercising human skeletal muscle.

Nitric oxide (NO) attenuates alpha-adrenergic vasoconstriction in contracting rodent skeletal muscle, but it is unclear if NO plays a similar role in human muscle. We therefore hypothesized that in humans, NO produced in exercising skeletal muscle blunts the vasoconstrictor response to sympathetic activation. We assessed vasoconstrictor responses in the microcirculation of human forearm muscle using near-infrared spectroscopy to measure decreases in muscle oxygenation during reflex sympathetic activation evoked by lower body negative pressure (LBNP). Experiments were performed before and after NO synthase inhibition produced by systemic infusion of N(G)-nitro-L-arginine methyl ester (L-NAME). Before L-NAME, LBNP at -20 mmHg decreased muscle oxygenation by 20 +/- 2 % in resting forearm and by 2 +/- 3 % in exercising forearm (n = 20), demonstrating metabolic modulation of sympathetic vasoconstriction. As expected, L-NAME increased mean arterial pressure by 17 +/- 3 mmHg, leading to baroreflex-mediated suppression of baseline muscle sympathetic nerve activity (SNA). The increment in muscle SNA in response to LBNP at -20 mmHg also was attenuated after L-NAME (before, +14 +/- 2; after, +8 +/- 1 bursts min(-1); n = 6), but this effect of L-NAME was counteracted by increasing LBNP to -40 mmHg (+19 +/- 2 bursts min(-1)). After L-NAME, LBNP at -20 mmHg decreased muscle oxygenation similarly in resting (-11 +/- 3 %) and exercising (-10 +/- 2 %) forearm (n = 12). Likewise, LBNP at -40 mmHg decreased muscle oxygenation both in resting (-19 +/- 4 %) and exercising (-21 +/- 5 %) forearm (n = 8). These data advance the hypothesis that NO plays an important role in modulating sympathetic vasoconstriction in the microcirculation of exercising muscle, because such modulation is abrogated by NO synthase inhibition with L-NAME.

Adult↗

Effects of heroin and its metabolites on schedule-controlled responding and thermal nociception in rhesus monkeys: sensitivity to antagonism by quadazocine, naltrindole and beta-funaltrexamine.

Recent studies have reported differences in the receptor mechanisms and intrinsic efficacies of heroin and its metabolites 6-acetylmorphine and morphine in rodents. The present study examined the generality of these findings to rhesus monkeys using two behavioral procedures. In an assay of schedule-controlled behavior, response rates were recorded under a fixed-ratio 30 schedule of food presentation. In an assay of thermal nociception, tail-withdrawal latencies were measured from warm water (42-58 degrees C). Heroin, 6-acetylmorphine and morphine produced dose-dependent rate-decreasing and antinociceptive effects. Antagonism studies were conducted with the competitive mu-selective antagonist quadazocine, the competitive delta-selective antagonist naltrindole, and the irreversible mu-selective antagonist beta-funaltrexamine (beta-FNA). Quadazocine dose-dependently antagonized the effects of all three opioids. Quadazocine pA2 values were similar across drugs and assays (7.4-7.8) and similar to quadazocine pA2 values for antagonism of other mu agonists. In contrast, naltrindole did not alter the effects of any of the opioids. beta-FNA antagonized the rate-decreasing and antinociceptive effects of heroin and morphine. Dose-effect data for heroin- and morphine-induced antinociception alone and after beta-FNA treatment were used to estimate in vivo apparent efficacy values (tau). Tau values (95% confidence limits) were 8.1 (6.9-9.6) for heroin and 2.6 (2.5-2.9) for morphine, but this difference is relatively small. These results suggest that the rate-decreasing and antinociceptive effects of heroin, 6-acetylmorphine and morphine are mediated by pharmacologically similar populations of mu opioid receptors in rhesus monkeys. The in vivo apparent efficacy of heroin at mu receptors was similar to or only slightly greater than that of morphine.

Animals↗

Activation and control of self-reactive gammadelta T cells.

Mammalian and avian CD3+ T cells can be separated into two lymphocyte subsets bearing heterodimeric T-cell receptors (TCR) composed of either alphabeta or gammadelta chains. Although it is now widely accepted that gammadelta and alphabeta T cells fulfill mandatory and nonredundant roles, the generality of this assumption and the exact functions played by gammadelta T cells remain uncertain. While an early protective role of gammadelta T cells has long been suspected, recent observations drawn in particular from transgenic models suggest their implication in the homeostatic control of immune and nonimmune processes. This hypothesis is also supported by the existence of several self-reactive gammadelta T-cell subsets in rodents and humans, whose specificity and effector properties will be detailed and discussed here. The present review will also describe several mechanisms that could allow efficient control of these self-reactive subsets while permitting expression of their regulatory and/or protective properties.

Animals↗

Home environmental intervention in inner-city asthma: a randomized controlled clinical trial.

BACKGROUND: Airborne pollutants and indoor allergens increase asthma morbidity in inner-city children; therefore, reducing exposure, if feasible, should improve asthma morbidity. OBJECTIVE: To conduct a randomized controlled trial of methods to reduce environmental pollutant and allergen exposure in the homes of asthmatic children living in the inner city. METHODS: After the completion of questionnaires, spirometry and allergen skin tests, home inspection, and measurement of home air pollutant and allergen levels, 100 asthmatic children aged 6 to 12 years were randomized to the treatment group (home-based education, cockroach and rodent extermination, mattress and pillow encasings, and high-efficiency particulate air cleaner) or to the control group (treated at the end of the 1-year trial). Outcomes were evaluated by home evaluations at 6 and 12 months, clinic evaluation at 12 months, and multiple telephone interviews. RESULTS: In the treatment group, 84% received cockroach extermination and 75% used the air cleaner. Levels of particulate matter 10 microm or smaller declined by up to 39% in the treatment group but increased in the control group (P < .001). Cockroach allergen levels decreased by 51% in the treatment group. Daytime symptoms increased in the control group and decreased in the treatment group (P = .04). Other measures of morbidity, such as spirometry findings, nighttime symptoms, and emergency department use, were not significantly changed. CONCLUSIONS: A tailored, multifaceted environmental treatment reduced airborne particulate matter and indoor allergen levels in inner-city homes, which, in turn, had a modest effect on morbidity.

Air Pollution, Indoor↗

Animal models of diabetes and obesity, including the PBB/Ld mouse.

Diabetes mellitus occurs in many animals species. However, only a few have been utilized in systematic studies designed to answer unsolved problems associated with the disorder in man such as molecular basis, pathogenesis of the vascular and neural lesions, and the roles of diet, exercise and obesity. Among the animal models available, rodents have been studied most thoroughly for a number of reasons: a) short generation time (sexually mature at about 3 mo of age, gestation time 21 days) and life-span is approximately 3 yr; b) hyperglycemia and/or obesity is known to be inherited in several species; c) environmental factors can be controlled easily in the laboratory because of small size; and d) economic considerations. The better-known rodent diabetes/obesity syndromes may be categorized as follows: 1) hyperglycemic with ketoacidosis, nonobese (Chinese hamster, South African hamster); 2) hyperglycemic with insulin hypersecretion, moderate obesity and may develop ketoacidosis (diabetic mouse (db/db), spiny mouse, sand rat); and 3) less pronounced hyperglycemia with hyperinsulinemia, insulin "resistance" and marked obesity (obese (ob/ob), yellow (Ay) and New Zealand obese (NZO) mice, and the Zucker "fatty" rat). The PBB/Ld mouse, described here in detail for the first time, is a new strain of mouse that also fits into the latter category. Members of this strain following maturity develop an obesity that is characterized by increasing cellularity of adipose tissue, increased serum immunoreactive insulin, reduced glucose tolerance, fatty liver, and hyperlipidemia. Therefore, this strain of mouse represents another model for study of adult onset obesity.

Adipose Tissue↗

Cell proliferation rates in common cancer target tissues of B6C3F1 mice and F344 rats: effects of age, gender, and choice of marker.

Increasing emphasis is being placed on mode of action for chemical carcinogens as an important consideration for risk assessment. Many rodent carcinogens appear to act through nongenotoxic mechanisms, such as induced cell proliferation. Information on cell proliferation rates based on species, age, gender, tissue, and choice of marker will provide a foundation for incorporating such measurements into rodent toxicity studies. Cell proliferation was evaluated in liver, kidney, skin, and forestomach of control male and female B6C3F1 mice and F344 rats at 7, 10, 13, and 20 weeks of age. Proliferating cell nuclear antigen (PCNA), an endogenous cell proliferation marker, and bromodeoxyuridine (BrdU) administered by ip injection 2 hr before euthanization were compared as markers of cell proliferation. Only in liver were BrdU and PCNA labeling indices (LIs; S phase only) statistically similar. As expected, the PCNA proliferating index (PI; G1 + S + G2 + M phases) was consistently greater than the S phase LI in all tissues examined. Age-related differences in LI were evident in liver and kidney, whereas LIs in the forestomach and skin were not age- dependent. In all tissues examined, gender- and species-related differences in cell proliferation were detected. Although BrdU and PCNA LIs were often statistically different, they both provided a useful indication of cell proliferation rates in the tissues examined. These results provide potentially useful information for designing rodent toxicity studies and biological models of carcinogenesis.

Age Factors↗

Zinc supplementation decreases hepatic copper accumulation in LEC rat: a model of Wilson's disease.

The effect of dietary zinc (Zn) supplementation on copper (Cu)-induced liver damage was investigated in Long-Evans Cinnamon rats (LEC), a model for Wilson's disease (WD). Four-week-old LEC (N=64) and control Long-Evans (LE) (N=32) female rats were divided into two groups; one group was fed with a Zn-supplemented diet (group I) and the other was given a normal rodent diet (group II). LEC rats were killed at 6, 8, 10, 12, 18, and 20 wk of age; the LE control rats were killed at 6, 12, 18, and 20 wk of age. Cu concentration in the liver was reduced in LEC rats fed the Zn-supplemented diet compared with LEC rats on the normal diet between 6 and 18 wk of age. Metallothionein (MT) concentration in the livers of LEC rats in group I increased between 12 and 20 wk of age, whereas hepatic MT concentration in LEC rats from group II decreased after 12 wk. Hepatocyte apoptosis, as determined by TUNEL, was reduced in Zn-supplemented LEC rats at all ages. Cholangiocellular carcinoma was observed only in LEC rats in group II at wk 20. These results suggest that Zn supplementation can reduce hepatic Cu concentration and delay the onset of clinical and pathological changes of Cu toxicity in LEC rats. Although the actual mechanism of protection is unknown, it could be explained by sequestration of dietary Cu by intestinal MT, induced by high dietary Zn content.

Animals↗

Decreases in spontaneous tumors in rats and mice after treatment with amphetamine.

Toxicology and carcinogenesis studies of dl-amphetamine sulfate, a drug used in the treatment of weight control, narcolepsy, and behavioral syndromes in children, were performed in F344/N rats and B6C3F1 mice. In these studies, amphetamine was administered for 2 years at doses of 0, 20, or 100 ppm in the feed to groups of 50 animals/dose/sex/species. The average amount of amphetamine consumed per day was estimated to be 1 or 5 mg/kg for low or high dose rats, 4 or 30 mg/kg for low or high dose male mice, and 3 or 19 mg/kg for low or high dose female mice. Survival was similar in dosed and control groups. The most notable effect of long-term treatment with this drug was the reduction of body weight in comparison to controls, and reduction in spontaneous tumors including pheochromocytomas of the adrenal gland in male rats, fibroadenomas of the mammary gland in female rats, adenomas of the anterior pituitary gland in male and female rats and female mice, endometrial stromal polyps of the uterus of female rats, adenomas or carcinomas of the liver in male and female mice, adenomas of the Harderian gland in male and female mice, and adenomas or carcinomas of the lung in male and female mice. Decreases in spontaneous tumors have previously been seen in 2-year rodent studies in groups of animals that have a reduced body weight in comparison to controls, but the spectrum of reduction in spontaneous neoplasms after treatment with amphetamine is broader than has previously been observed.

Adrenal Gland Neoplasms↗

Effects of gonadal hormones on central nitric oxide producing systems.

Nitric oxide-containing neurons are widely distributed within the CNS, including regions involved in the control of reproduction and sexual behavior. The expression of neuronal nitric oxide synthase is influenced by testosterone in male rat, and by estrogens in female. Moreover, nitric oxide synthase may co-localize with gonadal hormones' receptors. Gonadal hormones may influence nitric oxide synthase expression in adulthood as well as during the development. In fact, in mice knockout for estrogen receptor alpha, the nitric oxide synthase-expressing population is deeply reduced in specific regions. In physiological conditions, the female in mammalian species is exposed to short-term changes of gonadal hormones levels (estrous cycle). Our recent studies, performed in the rat vomeronasal system and in mouse hypothalamic and limbic systems reveal that, in rodents, the expression of nitric oxide synthase-producing elements within regions relevant for the control of sexual behavior is under the control of gonadal hormones. The expression of nitric oxide synthase may vary according to the rapid variations of hormonal levels that take place during the estrous cycle. This seems in accordance with the hypothesis that gonadal hormone activation of nitric oxide-cyclic guanosine-monophosphate pathway is important for lordosis behavior, as well as that this system is activated during mating behavior. Finally, comparative data available for other vertebrates suggest that class-specific and species-specific differences occur in the nitric oxide synthase system of hypothalamus and limbic structures. Therefore, particular caution is needed to generalize data obtained from studies in rodents.

Androgens↗

mRNA quantification of C-type natriuretic peptide in brain areas of rodents.

C-type natriuretic peptide (CNP) is regarded as an endothelium-derived vasodilator and might therefore have an important role in controlling vascular tone and remodeling. Because CNP also is expressed in the brain, it is considered to be a neurotransmitter. The present study compares expression levels of CNP mRNA in distinct areas of the mouse brain with the expression pattern in the rat brain. A distinct expression of CNP was found in all investigated areas with the exception of the mouse striatum. In both rodents, high CNP expression was detected in the tegmentum.

Animals↗

ELISA assay for IgG autoantibody to G-actin: comparison of chronic active hepatitis and acute viral hepatitis.

An ELISA was developed to test for IgG autoantibody to rabbit skeletal muscle G-actin in liver disease. To express results of the ELISA, we used a single reference serum with known high anti-actin activity, and assessed binding of test sera as a percentage of that displayed by the reference antiserum. Antibody to G-actin was measured in 40 sera from patients with chronic active hepatitis (CAH), 39 from acute viral hepatitis A (Hep A), 46 from acute viral hepatitis B (Hep B), 23 from non-A, non-B hepatitis (Hep non-A, non-B), 31 from systemic lupus erythematosus (SLE), 21 from scleroderma and 93 from normal persons. The results were compared with those obtained using indirect immunofluorescence tests with frozen sections of rodent stomach or fibroblast monolayers as substrates. Anti-G-actin activity of serum was significantly higher (P less than 0.001) in CAH (mean 63% +/- 23, range 18-110) than in controls (mean 15% +/- 12, range 0-51), systemic lupus erythematosus (mean 25% +/- 16, range 8-76), scleroderma (mean 23% +/- 13, range 3-59), Hep A (mean 24% +/- 11, range 6-25), Hep B (17% +/- 8, range 2-36), or Hep non-A, non-B (mean 23% +/- 11, range 7-53). There was no significant difference (P greater than 0.05) in the occurrence of anti-G-actin activity of serum in Hep A, Hep B or non-A, non-B compared with controls. At an ELISA reading of greater than or equal to 40% (mean + 2 s.d. of controls) the assay detected autoantibodies to G-actin in 85% of CAH, compared to 80% on fibroblast monolayers and 70% on rodent stomach. The ELISA described in the present study is a simple, sensitive and quantitative assay for autoantibody to G-actin. It should prove useful in assessing subspecificities of actin antibodies in liver diseases, and in differential diagnosis, particularly CAH from acute viral hepatitis.

Actins↗

Changes in the profiles of rodent plasma lipoproteins and apolipoproteins after cholesterol feeding.

Profiling of plasma lipoprotein distribution and apolipoprotein analysis were carried out semi-quantitatively by strictly controlled gel-filtration and SDS slab-gel electrophoresis for rats (Sprague-Dawley rats and ExHC rats), mice and hamsters. The effects of cholesterol feeding on lipoprotein and apolipoprotein patterns of these animals were also studied. HDL were major lipoproteins in all the animals. Hamsters had the most abundant lower density lipoproteins among the rodents with the ratio of cholesterol content between high density lipoproteins and lower density lipoproteins being about 1:1. After cholesterol feeding, the cholesterol content of lower density lipoproteins increased in all the animals. The increase was the greatest in ExHC rats and the least in mice. The profiling methods employed should be of value in determining the changes in the lipoprotein and apolipoprotein patterns of both experimental animals and man.

Animals↗

Clustered cases of leptospirosis in Rochefort, France, June 2001.

Five clustered cases of leptospirosis were diagnosed in the area of Rochefort, France, in June 2001, among teenagers who had swum in the Genouillé canal. The symptoms included fever, headache, abdominal pain and vomiting, chills and myalgia. Three cases were confirmed by PCR and serology. The mean cumulative duration of bathing was significantly higher in cases (23.8 hours) compared to controls (14.4 hours). No other particular risk factor was observed. The environmental investigation revealed the presence of rodents excreting of leptospires near the bathing area. For all antigens considered, the occurence of seropositive rodents was 30.8%, L. icterohaemorrhagiae being the predominant serogroup (23,1%).

Adolescent↗

Human melanoma proteoglycan: expression in hybrids controlled by intrinsic and extrinsic signals.

Human malignant melanoma cells express specific chondroitin sulfate proteoglycans (mel-CSPG) on the surface, both in vivo and in vitro. Melanocytes in normal skin show no detectable mel-CSPG but can be induced to express the antigen when cultured in the presence of cholera toxin and the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. Most other cell types do not express mel-CSPG either in vivo or in vitro. A study was designed to examine regulatory signals controlling mel-CSPG expression. The gene encoding mel-CSPG was mapped to human chromosome 15, and this chromosome was introduced into rodent cells derived from distinct differentiation lineages. Three types of mel-CSPG--expressing hybrids were found: (i) hybrids derived from human melanomas; (ii) hybrids derived from human cells that do not express mel-CSPG; and (iii) hybrids derived from human cells expressing mel-CSPG that are antigen-negative but that are induced to express mel-CSPG when cultured on extracellular matrix instead of plastic surfaces. Thus, mel-CSPG expression can be controlled both through intrinsic signals, provided by the differentiation program of the rodent fusion partner, and through extrinsic signals, provided by specific cell-matrix interactions.

Aggrecans↗

Adenovirus type 12 E1A down regulates expression of a transgene under control of a major histocompatibility complex class I promoter: evidence for transcriptional control.

The E1 region of human adenovirus type 12 (Ad12 E1) represses the expression of major histocompatibility complex (MHC) class I genes in transformed primary rodent cells. Conflicting results have been reported as to whether this E1A-mediated repression occurs at a transcriptional or a posttranscriptional level. In the present study, we show that in Ad12 E1-transformed primary baby mouse kidney cells from transgenic mice harboring the human growth hormone gene under control of an H-2K promoter both the expression of the endogenous MHC class I genes and the expression of the transgene are repressed. This experiment, as well as nuclear run-on analyses performed with single-stranded probes, revealed that Ad12 E1A inhibits MHC class I gene expression by repressing its promoter.

Adenovirus Early Proteins↗

Characterization of a second RNA-binding protein in rodents with specificity for iron-responsive elements.

Iron regulatory factor (IRF) is a cytoplasmic RNA-binding protein involved in regulating iron homeostasis. IRF controls expression of ferritin and transferrin receptor post-transcriptionally via specific binding to stem-loop iron-responsive elements (IREs) located in the untranslated regions of the respective mRNAs. We have confirmed by RNA band-shift analysis that a second IRE-protein complex observed in different rodent cell extracts is, like IRF, regulated by intracellular iron levels. This faster migrating complex appears to represent a specific interaction between the ferritin IRE and an iron-regulated protein that is distinct from IRF, as concluded from the following lines of evidence. First, UV cross-linking and partial digestion with different proteases revealed different peptide patterns for the two IRE-protein complexes. Second, antiserum raised against IRF peptides immunoprecipitated only authentic IRF and not the protein of the faster migrating complex, as determined by band-shift analysis. Following separation of the two IRE-binding proteins by ion-exchange chromatography, only the IRF-containing fraction reacted with the antibodies on Western blots. The second protein binds IREs with an affinity similar to that of IRF as demonstrated by competition with a ferritin IRE and related stem-loop RNAs. UV cross-linking experiments indicate that this second protein, tentatively named IRFB, has a molecular mass of approximately 105 kDa. Analysis of mouse tissues revealed differences in the distribution of IRF and IRFB. Whereas IRF protein and IRE binding activity were predominant in liver, intestine, and kidney, the IRFB protein(s) revealed highest binding activity in intestine and brain. Our data support the existence of two distinct iron-regulated IRE-binding proteins in rodents.

Animals↗