Analysis of the sheep MHC using HLA class I,II, and C4 cDNA probes

Four cDNA probes for the human major histocompatibility complex (MHC) were used to investigate the sheep MHC, in conjunction with serological typing for ovine lymphocyte antigen (OLA). Lymphocytes from a family (two parents and five offspring) of Romanov sheep were subjected to genomic DNA digestion by the restriction endonuclease Eco RI, followed by gel electrophoresis. A single Southern blot representing all seven individuals was then consecutively hybridized with the class I, alpha-DC, beta-DR, and C4 probes, which were originally designed to identify HLA class I, class II (DC and DR), and C4 products, respectively. Using each of the three class I/class II probes, several bands showing DNA polymorphism were detected. The segregation of these bands in the five offspring exactly paralleled the OLA haplotype segregation established by serological typing. A further eight individuals carrying haplotypes which were phenotypically identical to those in the above-mentioned family showed bands in the corresponding positions when tested with the same three probes. Using the C4 probe, no polymorphism was detected in these fifteen individuals.Abbreviations used in this paper MHC major histocompatibility complex - OLA ovine lymphocyte antigen - kbp kilobase pair(s) - MLR mixed lymphocyte reaction - RFLP restriction fragment length polymorphism     

Altered central monoamine response tod-amphetamine in rats chronically exposed to inorganic lead

Investigations of the mechanisms involved in the neurotoxicity resulting from chronic inorganic lead (Pb) exposure have centered on CNS biogenic amine function on the basis of behavioral and neurochemical findings. The following study examined the time course of the response of dopamine (DA) and 5-hydroxytryptamine (5-HT) neurons to d-amphetamine (AMPH) in rats chronically exposed to Pb from birth in order to further examine neurochemical mechanisms implicated by previous work. Offspring were exposed to 0.2% Pb acetate via the lactating dam and then weaned to the same drinking solution. At 120–140 days animals were injected with 1.0 mg/kg s.c. of the drug or with saline and sacrificed after various intervals. DA content in nucleus accumbens and corpus striatum in Pb-exposed animals was significantly higher than corresponding levels in controls at 20 minutes post-drug and remained significantly higher than baseline values at 80 minutes after the drug when DA concentrations in controls had returned to normal. These data suggest enhanced AMPH-induced DA synthesis in exposed rats. 5-Hydroxyindoleacetic acid (5-HIAA) content was significantly increased in three brain regions in exposed rats given AMPH compared to values in saline-injected exposed animals, indicating a compensation in these areas for the decreases in 5-HIAA values produced by Pb exposure alone. The results of this study reinforce the hypothesis that DA and 5-HT neurons are sensitive to relatively low levels of Pb exposure.     

Protoplast transformation of group N streptococci with cryptic plasmids

Abstract By using an extension to group N streptococci of a contransformation procedure we have introduced 4 different-sized cryptic plasmids for Streptococcus lactis into the plasmid-free S. lactis IL1403. A mixture of 4 cryptic plasmids with an indicator plasmid (pHV1301) conferring erythromycin resistance was used for IL1403 protoplast transformation. Under such conditions, 41.5% of the erythromycin-resistant transformants were contransformed with one of the cryptic plasmids in addition to pHV1301. Indicator plasmid pHV1301 was later spontaneously segregated from doubly transformed cells. This protocol should be very useful for constructing lactic streptococcal strains bearing any phenotypically cryptic plasmid.     

Thermophilic anaerobic spirochetes in New Zealand hot springs

Abstract Electron and light microscopy revealed the presence of spirochetes in New Zealand thermal springs. The spirochete population in one spring studied (Kuirau Lake) was affected by fluctuations in temperature and/or pool level. A pure culture of the strictly anaerobic bacterium revealed that it grew optimally at a temperature of 45–50°C, with no growth occurring above 60°C, and a pH of 7.0–7.5 with no growth occurring at pH 5.5 or 8.5. Growth was inhibited by chloramphenicol, penicillin, streptomycin, tetracycline and neomycin but not by d -cycloserine, novobiocin or phosphomycin at 10 μg/ml. A wide range of carbohydrates were utilized but not organic acids. Acetate was the major end product of glucose fermentation with substantial amounts of ethanol and traces of lactate being produced.     

Comparison of a Thermoanaerobium sp. from a New Zealand hot spring with Thermoanaerobium brockii

Abstract An anaerobic ethanologenic strain of extremely thermophilic bacteria isolated from a New Zealand hot spring resembled Thermoanaerobium brockii in morphology and cell-wall ultrastructure. However, antibodies produced against the New Zealand isolate did not crossreact with the type strain of T. brockii . The New Zealand isolate strain Tok6-B1 fermented a wider range of carbohydrate substrates, including pentoses, and was less inhibited by a hydrogen atmosphere. Ethanol and acetate were major end-products and lactate a minor product of glucose fermentation. Under a hydrogen atmosphere, these 3 end-products were formed in approximately equal amounts.     

Effects of particle concentration and season on the filtration rates of the freshwater clam,Sphaerium striatinum Lamarck (Bivalvia: Pisidiidae)

The effects of particle concentration and season on the filtration rates of the freshwater clamSphaerium striatinum Lamarck were assessed by measuring clearance rates of small (2.02 μm) latex beads from dilute suspensions. Filtration rates decreased as particle concentration increased over a range of 2–64 mg 1⁻¹, with rates decreasing in similar proportion for clams of all sizes. For a 1-mg clam, rates decreased from approximately 8.4 to 0.57 ml clam⁻¹ h⁻¹. Seasonal filtration rates for adult clams peaked during periods of greatest reproduction. The patterns for smaller clams are similar, though proportional changes in filtration rates differ for various sizes of clams. It is estimated that clams occupying 1 m² of stream substrate removed about 3.67 gCa⁻¹. This is equivalent to 0.0004% of the carbon that flows past them annually. Filter-feeding provided only 24% of the calculated energy needs of the population, suggesting that another mode of feeding (e.g. deposit-feeding) may provide an important energy source for these forms. Funded in part by a grant-in-aid to D. J. Hornbach from Sigma-Xi, The Research Society. Funded in part by a grant-in-aid to D. J. Hornbach from Sigma-Xi, The Research Society.     

Determining the density dependence of immigration and emigration of benthic stream invertebrates: theoretical considerations

Simple mathematical models are formulated to describe density independent and density dependent dispersal. These models clarify hypotheses of density dependence and may be manipulated easily to suit particular applications. The models demonstrate that the initial composition of a species aggregate must be controlled before valid conclusions can be drawn about the density dependency of the aggregate's dispersal. Stochastic models of emigration are derived to assess the power of particular experimental designs and statistical techniques to discriminate a known form of density dependent emigration. Contribution No. 369, Great Lakes Research Division, University of Michigan Contribution No. 369, Great Lakes Research Division, University of Michigan     

Prevention of stress-induced gastric ulcers by dopamine agonists in the rat

Dopamine (DA) and DA agonists have been shown to exert a protective role against the formation of duodenal ulcers. The effect of stimulation of DA receptors on the development of stress-induced gastric ulcers is currently unknown. Accordingly, we evaluated the effect of several DA agonists on the development of gastric ulcers induced by 3 h of cold + restraint stress (CRS) in rats. Apomorphine, d-amphetamine, methylphenidate, and threo-dl-p-hydroxymethylphenidate (an hydroxylated analog of methylphenidate), significantly reduced both the incidence and severity of CRS-induced gastric ulcers. The gastric cytoprotection afforded by these agents was dose-related, and completely antagonized by pretreatment with the peripheally acting DA antagonist domperidone. Because domperidone blocks peripheral, but not central, DA receptors, and since the entry of threo-dl-p-hydroxymethylphenidate across the blood-brain barrier into the brain is restricted to a great extent, we conclude that stimulation of peripheral DA receptors is primarily involved in the gastric cytoprotection induced by dopamimetics.The pathogenesis of stress-induced gastric ulcers remains largely unknown, and significant efforts have been made over the last decade to functionally characterize some of the factors involved in the etiology of this disease. Considerable attention has been focused on gastric acid secretion, but its primary role in stress-induced gastric ulcer disease remains uncertain. In fact, agents which effectively inhibit or neutralize gastric acid secretion such as cimetidine or antacids do not necessarily exert protection against stress-induced gastric ulcers (1,2). Moreover, in our original studies with neurotensin, a brain and gastrointestinal peptide, we have found that central administration of this neuropeptide, which completely prevents the development of cold + restraint stress (CRS)-induced gastric ulcers, does not appreciably alter gastric acid secretion (2). These findings support the contention that gastric acid secretion may not be an important factor in the development of this type of gastric ulcer.There is, however, considerable evidence that the automatic nervous system plays an intermediary role in the development of these ulcers (3,4). In this regard, surgical or pharmacological blockade of the vagal (cholinergic) division of the autonomic nervous system prevents the appearance of stress-associated gastric ulcers (5,6). Direct stimulation of catecholamine receptors, or indirect activation via increased sympathetic outflow to the periphery (7,4,8–11) appears to produce a salutary effect of stress-induced gastric ulcers.Szabo and his associates (12, 13, 14) have extensively studied the anti ulcer effects of dopamine (DA) in duodenal ulcer formation. Whether DA also modifies the development of stress-induced gastric ulcers is currently unknown.We have therefore evaluated the effect of selected DA receptor agonists and antagonists on CRS-induced gastric ulcer formation in rats.     

Theophylline delays human sleep phase

Twelve healthy male volunteers were given theophylline 250 mg in order to test effects on 24-hr rhythms. Rhythms of sleep/wake and subjective sleepiness were delayed. Ingestion of xanthines such as theophylline in coffee, tea, colas and chocolate may contribute to some sleep disorders. Theophylline might likewise be useful in treating disorders of circadian oscillators.     

Histopathological and ultrastructural changes in the antennal gland,midgut, hepatopancreas,and gill of grass shrimp following exposure to hexavalent chromium

Grass shrimp, Palaemonetes pugio, were exposed for 1 month to subacute concentrations of hexavalent chromium (0.5, 1.0, 2.0, 4.0 ppm) after which the gills, midgut, hepatopancreas, and antennal glands were examined for histopathological and ultrastructural changes. Pathological changes were greatest in the antennal glands, followed by hepatopancreas, gills, and midgut. Severe changes occurred in some shrimp, even at 0.5 ppm chromium. Cells of all tissues frequently had both swollen mitochondria and rough endoplasmic reticulum. Small, spherical or ring-like intranuclear inclusions, possibly indicative of cellular hyperactivity or manifestions of chromium and/or protein complexes, were most prevalent in the hepatopancreas and antennal glands but also occurred in the midgut and gills. Other major degenerative changes in the antennal glands were restricted to the labyrinth and included diminution of basal plasmalemmal infoldings and cytoplasmic density, nuclear hypertrophy followed by widespread nuclear pyknosis and epithelial desquamation. In severely altered hepatopancreas hypertrophy was indicated for the basal laminae, nuclei, and possibly for the nucleoli. There was an apparent reduction in mitotic events and many observed mitotic nuclei were abnormal. Abnormal midgut hypertrophy was present in only 8 of 20 examined shrimp, exposed to 0.5 and 1 ppm chromium. Further, the gills of only 10 of the 40 examined chromium-exposed shrimp possessed abnormal features detectable with ligh microscopy. Ultrastructural analysis of the latter indicated an increase in lysosomes and a decrease in cytoplasmic density. In addition, there was a pronounced diminution in the degree of lamellar, subcuticular plasmalemmal infolding. This latter feature is postulated to be a mechanism for the regulation of chromium influx. Possible explanations for most observed alterations in the above tissues are proposed.