Endotoxic activity of cell-free rumen fluid from cattle fed hay or grain

The cell-free rumen fluid from cattle fed hay or grain exhibited the following biological characteristics which strongly suggest the presence of endotoxin or a toxic principle similar to endotoxin of gram-negative bacteria: proved lethal to mice when injected with actinomycin D; proved extremely lethal to chick embryo; induced biphasic pyogenic response in rabbits; enhanced susceptibility to bacterial infection in mice; evoked positive epinephrine skin reaction in rabbits and phenol-water or aqueous ether proved lethal to mice and chick embryos. A quantitative difference in concentrations of endotoxin was observed on LD50 in mice and chick embryos and response to the epinephrine skin test in rabbits. Cell-free rumen fluid of grain-fed cattle contained at least twice as much endotoxin as that of hay-fed cattle. Endotoxin in cell-free rumen fluid and in higher concentration in cattle fed grain than in those fed hay support the hypothesis that rumen bacterial endotoxins may participate in the pathogenesis of diseases associated with high grain feeding such as lactic acidosis and the sudden-death syndrome.     

S-adenosylmethionine metabolism and DNA methylation in hydrazine-treated rats

The treatment of rats with hepatotoxic doses of hydrazine (NH2-NH2) induces the rapid formation of 7-methylguanine and O6-methylguanine in liver DNA. The methyl moiety in these reactions might be derived from the cellular S-adenosylmethionine pool because radioactivity administered to these rats as methionine rapidly appears in the DNA as methylated guanine. An increased incorporation of radioactivity into 5-methylcytosine was previously reported followed by subsequent suppression. This increased radiolabeling of 5-methylcytosine coincided with time of maximal DNA guanine methylation. To determine the nature of S-adenosylmethionine metabolism during the period of DNA methylation induced by hydrazine treatment, and to determine if the increased radiolabeling of 5-methylcytosine at this time reflected an actual increase in 5-methylcytosine synthesis, liver DNA synthesis and S-adenosylmethionine levels and turnover were assayed. Liver S-adenosylmethionine concentrations varied slightly between control rats and hydrazinetreated rats during the first five hours after hydrazine administration, and no difference was detectable in the incorporation of administered [3H]methionine into S-adenosylmethionine. Because S-adenosylmethionine specific radioactivity in hydrazine-treated rats was not different from control rats, the previously observed increased radiolabeling of 5-methylcytosine appeared to represent an actual increase in synthesis. This conclusion was supported by finding that incorporation of radioactive thymidine into DNA was also accelerated immediately following hydrazine administration, again followed by a decrease. 5-Methylcytosine sythesis, therefore, appears to follow DNA synthesis during hydrazine toxicity, and formation of 7-methylguanine and O6-methylguanine in liver DNA of hydrazine-treated rats occurs during a short period of increased DNA sythesis and 5-methylcytosine formation very early in hydrazine toxicity.     

Air caloric test: irrigation technique

The technique of using air to perform the caloric test is discussed. Greater temperature differences and larger volumes of air are needed to achieve stimulations equal to those caused by water irrigations. Special consideration must be given to the design and operation of air irrigation equipment. The effects of irrigating a wet canal and of misdirecting the air jet are discussed. A new air irrigation apparatus is described.     

Oral keratinocyte immune responses in HIV-associated candidiasis

Technetium-99m is the most commonly used radionuclide in routine nuclear medicine imaging procedures. Development of 99mTc-labeled receptor-specific imaging agents for studying the central nervous system is potentially useful for evaluation of brain function in normal and disease states. A novel 99mTc-labeled tropane derivative, [99mTc]TRODAT-1, which is useful as a potential CNS dopamine transporter imaging agent, was evaluated and characterized. After i. v. injection into rats, [99mTc]TRODAT-1 displayed specific brain uptake in the rat striatal region (striatum-cerebellum/cerebellum ratio 1.8 at 60 min), where dopamine neurons are concentrated. The specific striatal uptake could be blocked by pretreating rats with a dose of competing dopamine transporter ligand, beta-CIT (or RTI-55, i.v., 1 mg/kg). However, the specific striatal uptake of [99mTc]TRODAT-1 was not affected by co-injection of excess free ligand (TRODAT-1, up to 200 microg per rat) or by pretreating the rats with haloperidol (i.v., 1 mg/kg). The specific uptake in striatal regions of rats that had prior 6-hydroxydopamine lesion in the substantia nigra area showed a dramatic reduction. The radioactive material recovered from the rat striatal homogenates at 60 min after i.v. injection of [99mTc]TRODAT-1 showed primarily the original compound (>95%), a good indication of in vivo stability in brain tissue. Similar and comparable organ distribution patterns and brain regional uptakes of [99mTc]TRODAT-1 were obtained for male and female rats. Ex vivo autoradiography results of rat brain sections further confirmed the high uptake and retention of [99mTc]TRODAT-1 in the striatal region. In vitro binding studies measuring the affinity to dopamine transporters for the free ligand, TRODAT-1, and a nonradioactive rhenium derivative, Re-TRODAT-1, showed Ki values of 9.7 nM and 14.1 nM, respectively. Behavioral studies in rats using the free ligand, TRODAT-1 and Re-TRODAT-1 indicated that, unlike other tropane derivatives, they displayed no effect on locomotor activity, suggesting low toxicity. These results strongly support the conclusions that this novel 99mTc radioligand binds selectively to dopamine transporters in the brain and that is is potentially useful for in vivo assessment of the loss of dopamine neurons in Parkinson's and other neurodegenerative diseases.     

Nitric oxide synthase in neuroepithelial brain tumors

Immunohistochemical investigation of NO-synthase in brain astrocytic tumors revealed intense reaction in many tumor cells as well as direct correlation in the intensity of reaction and the degree of tumor anaplasia. Grade I astrocytomas did not show immunoreactivity in contrast to high anaplastic tumors where many cells had positive reaction with a different degree of intensity. Positive immunoreaction was shown in many giant cells. Small cell glioblastomas and oligodendrogliomas were immunonegative. There was a direct correlation between NO-synthase expression and glial fibrillar acidic protein.     

The effect of carotid sinus denervation on fetal heart rate variation in normoxia, hypoxia and post-hypoxia in fetal sheep

BACKGROUND: Previous reports documented high rates of tobacco use among Alaska Natives (Eskimos, Indians, and Aleuts). In this population, tobacco use is the leading preventable cause of death. Lung cancer is the leading cause of cancer death among Alaska Natives and tobacco is responsible for over one-third of all cancer deaths in this population. Until recently there has been no systematic surveillance of the prevalence of tobacco use in this high-risk population. Data that did exist were not readily available to those primarily responsible for the health care of this population. This is the first time since 1990 that data on Alaska Natives have been collected in one analysis; this permits a more representative evaluation of tobacco use. METHODS: Data on tobacco use were obtained and analyzed from national and state surveys and selected research projects from 1988 to 1993. RESULTS: Alaska Natives have high prevalence of tobacco use, including both cigarettes and smokeless tobacco. Tobacco use prevalence among Alaska Natives exceeds that of Alaska non-Natives, U.S. whites, and American Indians/Alaska Natives in the United States outside of Alaska. Smoking prevalence among Alaska Native women is twice that of non-Native women in Alaska and nearly twice as high among pregnant Alaska Natives than pregnant non-Natives. Overall, prevalence of smokeless tobacco use was four times higher among Alaska Natives than comparative state and national populations. CONCLUSION: Because this population has such high rates of tobacco use, it is important to public health that monitoring and educational programs be in place and that data specific to Alaska Natives be made available.     

Cyclodextrin-based carbohydrate clusters by amide bond formation

Per-6-amino-2,3-dimethyl-ß-cyclodextrin was prepared very efficiently as its hydrochloride salt from native ß-cyclodextrin in four steps and 89% overall yield. O-Acetyl-protected ß-D-thioglucose and ß-D-thiolactose derivatives, containing short spacer arms terminated with carboxylic acid functions, were prepared by the BF₃·OEt₂-catalyzed thioglycosylation of ß-D-glucose pentaacetate and ß-lactose octaacetate with 3-mercaptopropionic acid, respectively. Utilizing amide bond formation as the key step, these thio-ß-D-glucosyl and lactosyl derivatives were coupled to per-6-amino-2,3-dimethyl-ß-cyclodextrin to afford, after deprotection, perfunctionalized ß-cyclodextrin-based clusters containing seven thio-ß-D-glucosyl and seven ß-lactosyl appendages, respectively. Molecular modeling of both these ß-cyclodextrin-based clusters revealed the glucose and lactose clusters to be approximately 23 Å and 27 Å in diameter, respectively, and approximately 19 Å in height in both cases. The association constants for the complexation of the anti-inflammatory drug naproxen by ß-cyclodextrin, per-2,3-dimethyl-ß-cyclodextrin, and the lactose cluster of ß-cyclodextrin in 0.01 M phosphate buffered saline solution (pH 7.4) were found by UV-vis spectrophotometric titration to be 374 ± 75 M⁻¹, 351 ± 70 M⁻¹, and 165 ± 33 M⁻¹, respectively.