The effects of a liquid ethanol diet on nutritional status and fluid balance in the rat.

The liquid ethanol diet is a widely used method of ethanol administration. The purpose of this study was to evaluate fluid balance using a multitude of physiological parameters (electrolytes, osmolality, total serum proteins, fluid intake/output and body weight), during and after the introduction of liquid ethanol diet. Animals were randomized into four different dietary protocols (two control and two ethanol groups) and were placed in metabolic cages for 16 days. Serum electrolytes, as well as the above parameters, were measured before, during and 1 week after the introduction of 9% (v/v) ethanol-containing diet (Lieber-DeCarli: LD). After the first night on 9% (v/v) ethanol LD, animals had significantly decreased diet consumption, urine output and body weight. However, a major finding of this study was that, during the habituation phase, the electrolyte values remained within the normal range for rats and, in particular, serum sodium was not altered at any time point measured in this study. Based upon the findings from this study, it is recommended that body weight be carefully monitored as a measure of the animal's equilibration and physiological adaptation during the initiation of a liquid ethanol diet, since neither the serum sodium nor calculated osmolality values were changed. Our results also highlight the need to offer water to animals during the habituation phase of ethanol consumption. This is because ethanol rats that were offered water ad libitum lost less weight than groups that did not receive water ad libitum, despite consuming the same amount of LD diet.

Physiologic and behavioral assessment of rabbits immunized with Freund's complete adjuvant.

Although the use of Freund's Complete Adjuvant (FCA) has been discouraged for the production of polyclonal antibodies, little clinical evidence supports the belief that FCA necessarily affects the well-being of immunized rabbits. We designed the present study to determine whether immunization at multiple sites with small volumes of Freund's adjuvant affects rabbit well-being. We injected 18 female New Zealand White rabbits (six animals per group) with antigen in FCA, Freund's Incomplete Adjuvant, or physiologic saline in the following volumes and routes: 0.02 to 0.03 mL intradermally in each of 30 to 40 sites and 0.1 mL subcutaneously in each of two sites. The body weight, temperature, complete blood count, and behavior of the rabbits in the home cage, upon handling, and in an open field did not differ significantly among the immunization groups during the 7-week assessment period. Only the degree of induration around injection sites differed: as expected, FCA induced the greatest response at the injection sites, but the sites were neither ulcerative nor necrotic, nor did palpation of the sites induce any apparent discomfort to the rabbits. We conclude that FCA may be used safely and humanely in rabbits if small volumes are injected intradermally or subcutaneously in multiple sites.

Outbreak of Pasteurella pneumotropica in a closed colony of STOCK-Cd28(tm1Mak) mice.

Fifteen mice with Pasteurella pneumotropica orbital abscesses were noted in mice that were homozygous for a targeted Cd28 gene mutation. Only one mouse heterozygous for the Cd28 mutation was affected. According to phenotypic reactions and 16S rDNA sequencing, the isolates were most similar to biotype Heyl. This article provides evidence for an immunologic basis of susceptibility to P. pneumotropica infection. Fifteen mice with Pasteurella pneumotropica orbital abscesses were noted in mice that were homozygous for a targeted Cd28 gene mutation. Only one mouse heterozygous for the Cd28 mutation was affected. According to phenotypic reactions and 16S rDNA sequencing, the isolates were most similar to biotype Heyl. This article provides evidence for an immunologic basis of susceptibility to P. pneumotropica infection.

Verification of bacterial killing effects of cage wash time and temperature combinations using standard penicylinder methods.

We conducted the present study to evaluate various time and hot-water temperature combinations necessary to kill three common bacterial species in standardized cultures on stainless steel penicylinders, in accordance with methods approved by the Association of Official Analytical Chemists. Exposure for no more than 2 sec to water at 82.2 degrees C killed all three bacterial species, as did exposure for 3 sec to water at 80 degrees C, 4 sec at 77.8 degrees C, and 5 sec at 75.6 degrees C. We conclude that temperatures in the range of 75.6 degrees C to 82.2 degrees C will effectively kill vegetative bacteria in a matter of seconds and that failure to kill these bacteria in cagewash operations, with belt travel times in minutes rather than seconds, is due to other factors that prevent the effective application of sufficiently hot water to the bacterial load.

Endothelin-1-like immunoreactivity in a new rodent model of spontaneous hypertension.

The purpose of this study was to determine plasma and tissue endothelin-1 (ET-1)-like immunoreactivity in a new rodent model of spontaneous hypertension. Plasma and tissues were procured from pentobarbital-anesthetized 16- to 18-week-old male hamsters with spontaneous hypertension and genetically/age-matched normotensive hamsters. We found that ET-1-like immunoreactivity in the plasma was similar in both groups. However, renal and cardiac ET-1-like immunoreactivity was 11- and 1.7-fold higher in spontaneously hypertensive hamsters relative to normotensive hamsters, respectively (P < .05). ET-1-like immunoreactivity was slightly, but significantly, lower in the lung and spleen of spontaneously hypertensive hamsters relative to normotensive hamsters (P < .05). ET-1-like immunoreactivity in the liver and brain was similar in both groups. We conclude that ET-1-like immunoreactivity is significantly higher in two target organs for hypertension, kidney and heart, but not in plasma or brain of adult male hamsters with spontaneous hypertension, relative to genetically/age-matched normotensive hamsters. We suggest that renal and cardiac ET-1 could play a role in the natural history of spontaneous hypertension in hamsters.

Auditory brain stem response in young and old guinea pigs.

OBJECTIVE: To characterize age-related auditory changes in genetically similar guinea pigs. BACKGROUND: In humans and animals, changes in hearing are known to occur with age. METHODS: Brain stem-evoked responses were measured in genetically similar guinea pigs that ranged 6-36 months in age. Changes in hearing and the input/output function curve were determined. RESULTS: Threshold shift with increase in age was seen. Marked reduction in amplitude of response with increasing age was also demonstrated. No change was seen in latency or interpeak interval. CONCLUSIONS: In genetically similar guinea pigs, age-related changes in threshold occurred. Latency and interpeak intervals remained unchanged. Amplitude of response decreased substantially after 12 months of age to a greater extent than predicted by threshold shifts alone. This phenomenon appears important in understanding the pathophysiology of age-related hearing loss.

Initial characterization of hamsters with spontaneous hypertension.

The purpose of this study was to begin to characterize a new inbred strain of adult male hamsters with established spontaneous hypertension along with their genetically/age-matched normotensive controls. We found that mean arterial pressure was 162+/-3 mm Hg in hypertensive hamsters and 94+/-4 mm Hg in controls (mean+/-SEM; P<.05). Body weight was significantly lower in hypertensive hamsters relative to normotensive hamsters (P<.05). Hypertension was associated with a significant increase in heart weight, thickness of the left ventricular wall, and amplitude of the QRS complex in standard electrocardiographic leads I and aVR (P<.05). No gross or microscopic abnormalities were observed in other organs. Plasma renin activity and the number of circulating neutrophils were significantly increased in hypertensive hamsters relative to controls (P<.05). Serum concentrations of creatinine, blood urea nitrogen, sodium, potassium, and calcium as well as urinalysis were similar in both groups. Overall, these data suggest that the spontaneously hypertensive hamster could be a suitable model for the study of spontaneous hypertension.

Suppression of apoptosis by bcl-2 does not prevent p53-mediated control of experimental metastasis and anchorage dependence.

Mutations in the p53 tumor suppressor gene are frequently associated with the metastatic stage of tumor progression. Inactivation of p53 was shown to promote metastasis under experimental conditions. To determine the p53 functions that are involved in the control of tumor metastasis, we compared properties of three types of transformed mouse fibroblasts: with intact p53, with p53-mediated apoptosis suppressed by bcl-2 and with p53 inactivated by dominant negative mutants. Although expression of bcl-2 blocked apoptosis in detached cells and increased tumor cell survival in the blood circulation, it was insufficient to affect the ability of p53 to cause cell cycle arrest in detached cells and suppress experimental metastasis. For the suppression of metastasis complete inactivation of p53 was required. We conclude that the apoptotic function of p53 is dispensable for the p53-dependent suppression of experimental metastasis that is presumably achieved by controlling anchorage dependence. These data provide a possible explanation to dramatic differences in values of bcl-2 and mutant p53 as prognostic markers in human cancer.

Swollen eyelid associated with Foleyella sp infection in a chameleon.

An adult female Oustalet's chameleon was examined to determine the cause of a fluctuant enlargement of the right superior eyelid. Surgical exploration of the subcutaneous tissues of the eyelid revealed live microfilarial parasites, which were identified later as Foleyella sp. These parasites, although seldomly reported, are fairly common in imported chameleons and can be detected during examination of blood smears. Surgical removal continues to be the treatment of choice for these parasites, because the efficacy and safety of many new anthelmintic agents have not been determined for use in chameleons.

Comparisons of radioiodoestradiol blood-tissue exchange after intravenous or intraarterial injection.

PURPOSE: Our study determines and compares how the major organs of large animals handle exogenous halogenated bioactive sex steroids within the first minutes after their i.v. or i.a. injection. The rationale is that an understanding is needed of the acute physiological events because they affect decisions for how to optimize delivery of radiohalogenated sex steroid receptor ligands for purposes of medical imaging and modes of radiotherapy. METHODS AND MATERIALS: We used an indicator dilution technique that allows monitoring of blood-tissue exchange of radioactivity in a continuous manner in anesthetized surgically prepared swine. RESULTS: In swine, with by-passed liver circulation, the lungs allow the vast majority of [I-125]-16 alpha-iodo-17 beta-estradiol ([I-125]E) to be extracted from the blood perfusing the lung in the initial transit after i.v. injection in vivo. Similar outcome was observed for most major organs, including the CNS, intestines, spleen, peripheral appendages, and kidneys after i.a. injection of [I-125]E in vivo. However, within minutes the organs released the [I-125]E in its original chemical form back into the vascular system, with the exception of estrogen receptor (ER) rich tissues and the kidneys that retained the [I-125]E in its original form, although in the kidneys a nonpolar metabolite also accumulated. CONCLUSION: Our experiments confirm in a large animal model that radioiodoestradiol can be sequestered or concentrated in ER-rich sites. The liver and sex steroid receptor-rich organs modify considerably, by metabolism and sequestration, respectively, the acute distribution of bioactive steroids. Our data indicate potential for detection of ER in vivo in hormone-sensitive tumors, that is, in breast and endometrial cancers, and offer improved understanding of the recent studies in subjects with breast cancer that demonstrated that receptor imaging in vivo of steroid receptors with high-affinity radiolabeled ligands is possible in a clinical setting.

Evidence of a steroidogenic enzyme gene dose effect on adrenal gene expression in hereditary rabbit congenital adrenal hyperplasia.

We previously reported the gene deletion encoding cytochrome P-450 cholesterol side-chain cleavage enzyme (P-450SCC, resulting in complete elimination of the adrenal gene expression and causing congenital adrenal hyperplasia in the rabbit. Using the rabbit congenital adrenal hyperplasia model, we investigated the wild type (wt) P-450SCC gene dose effect on gene expression in three P-450SCC genotype animals [wt/wt, wt/mutant (mt), mt/mt] identified by Southern blot analysis. Northern blots using a rabbit P-450SCC cDNA probe revealed no detectable P-450SCC mRNA in individual adrenals of animals with congenital adrenal hyperplasia (mt/mt) and approximately half or slightly less than half the levels of the mRNA in the pooled adrenals of five heterozygous (wt/mt) newborn animals compared with the mRNA levels in the pooled adrenals of five homozygous normal (wt/wt) newborn animals. Identical P-450SCC mRNA levels were found individual adrenals of adult animals with regard to the P-450SCC genotype, although at a higher expression level than in the newborn animals of the same genotype. Control Northern blots using human CPY21-B cDNA and cytoplasmic actin cDNA probes confirmed the accuracy and integrity of RNA. Western immunoblotting using anti-ovine P-450SCC antibody revealed decreased P-450SCC protein in the adrenals of wt/mt animals at approximately half the level of the P-450SCC protein in the adrenals of the wt/wt animals. Baseline and ACTH-stimulated serum corticosterone (B) levels in vivo were similar between the age-matched wt/mt and wt/wt animals, whereas ACTH-stimulated B levels in adult animals were higher than those in the newborn animals irrespective of P-450SCC genotype.(ABSTRACT TRUNCATED AT 250 WORDS)

Chronic administration of flumazenil increases life span and protects rats from age-related loss of cognitive functions: a benzodiazepine/GABAergic hypothesis of brain aging.

Under barrier condition and with ad lib access to food and water, 20 Fischer-344 rats were chronically treated for 10 months with the benzodiazepine (BDZ) antagonist, flumazenil (FL; 4 mg/kg/day in drinking water acidified to pH = 3.0), beginning at the age of 13 months, while the group of 20 control age-matched rats received plain acidified water. The life span of the first 8 deceased rats treated with FL was significantly longer than that of the first 8 deceased rats in the age-matched control group. In tests for spontaneous ambulation and exploratory behavior in the Holeboard apparatus, conducted during the 3rd and the 8th month of treatment, the FL group, relative to controls, had significantly higher scores for the ambulation and exploratory behavior. In tests for unrewarded spontaneous alternation in the T maze, conducted at days 7, 39, 42, and 47 through 54 after drug withdrawal, i.e., at the age of 24-25 months, the FL-exposed group, compared to age-matched controls, showed a significantly higher percent of alternating choices, a behavior that was statistically comparable to that of the "young" 6-month-old controls. In the Radial Maze tests conducted 2 months after drug withdrawal, the FL group made significantly less "working memory" errors and "reference memory" errors, relative to the age-matched 25-month-old control group, a performance that was comparable to that of the young 7-month-old control group. In conclusion, chronic FL significantly protected rats from age-related loss of cognitive functions. It is postulated that the age-related alterations in brain function may be attributable to the negative metabolic/trophic influences of the "endogenous" benzodiazepine (BDZ) ligands and/or those ingested with food. A BDZ/GABAergic hypothesis of brain aging has been formulated which assumes that age-related and abnormally strong BDZ/GABAergic influences promote neurodegeneration by suppressing trophic functions of the aminergic and peptidergic neurons through opening of chloride channels in soma membrane and axon terminals, causing excessive hyperpolarizing and depolarizing inhibition, respectively. The review of human clinical and animal data indicates that FL has nootropic actions by enhancing vigilance cognitive and habituation processes.

A comparison of euthanasia methods in rats, using carbon dioxide in prefilled and fixed flow rate filled chambers.

The two methods (prefilled and fixed flow rate filled chambers) recommended in the 1993 AVMA Euthanasia Panel report for using carbon dioxide to euthanatize rats were evaluated in terms of their effect on behavior and selected blood gas values. Responses were videotaped during exposure to > or = 90% carbon dioxide in a prefilled chamber or a gradually filled chamber, using a fixed flow rate of 20% chamber volume/min. Arterial blood samples were taken to determine partial pressure of oxygen, partial pressure of carbon dioxide, pH, and oxygen saturation prior to entering the chamber and at time points determined by rats' responses to carbon dioxide. Rats showed similar reactions when exposed to carbon dioxide by either method. Significant differences in mean time for each response to occur were seen between euthanasia methods. Maintaining a near atmospheric oxygen chamber concentration by using a 75% CO2: 20% O2: 5% N2 gas mixture to gradually fill the chamber did not change rats' reactions upon exposure. Significant differences were found between pre-exposure values and values from samples obtained when rats became immobile after entering the prefilled chamber. Partial pressure of carbon dioxide significantly increased, and pH and percent oxygen saturation significantly decreased from pre-exposure values in all samples obtained after rats entered the gradually filled chamber. Partial pressure of oxygen in these rats was greater than or equal to pre-exposure levels in all samples. Rats appeared sedated because of the anesthetic effects of carbon dioxide when immobility was observed. Distress was not observed in the rats when either method of euthanasia was used.