The oral assimilation of radiomanganese by the mouse

The metabolism of orally administered radiomanganese was studied in mice. Assimilation of absorbed manganese (Mn) was determined using whole body counting techniques. When⁵⁴MnCl₂ was administered, 2.7% of the dose was retained after 10 d compared with 1.2% from the⁵⁴Mn-nitrilotriacetate (NTA) complex. However, this difference was accounted for by the rapid and persistent adsorption of the Mn onto the teeth of the lower jaw when fed as the ionic salt at pH 2.0 compared with the NTA-chelate fed at pH 9.0. Once corrected for the amount adsorbed onto the teeth, the biodistribution and relative specific activity of the assimilated radiomanganese into a variety of tissues were similar for both forms of the metal.     

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     

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.     

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.     

Paramecium secretory granule content: quantitative studies on in vitro expansion and its regulation by calcium and pH

Ca2+-dependent secretion in Paramecium involves the exocytic release of a paracrystalline secretory product, the trichocyst matrix, which undergoes a characteristic structural change from a highly condensed storage form (Stage I) to an extended needle-like structure (Stage III) during release. We studied trichocyst matrix expansion in vitro to examine factors regulating the state of secretory organelle content. A new method for the isolation of membrane-free, condensed (Stage I) trichocyst matrices is described. These highly purified, condensed matrices were used to develop a rapid quantitative, spectrophotometric assay for matrix expansion to examine factors regulating the Stage I and Stage III transition. Expansion from Stages I to III was elicited in vitro by addition of Ca2+ and we found that at neutral pH, expansion required a Ca2+ concentration slightly above 10(-6)M. Previous studies indicate that calmodulin (CaM) antagonists inhibit matrix expansion in vivo. However, in vitro matrix expansion is normal even when trichocyst matrices are preincubated in CaM antagonists before stimulation. Thus, matrix components themselves are unlikely to be the site of CaM antagonist action in vivo. In vitro matrix expansion is also modulated by pH. Decreasing pH to 6.0 inhibits expansion, i.e., expansion requires higher Ca2+ concentration. Conversely, increasing pH to greater than 7.0 promotes expansion, allowing it to occur at a lower Ca2+ concentration. The pH sensitivity of the Ca2+ binding sites of the matrix suggests that, in vivo, the interior of the trichocyst vesicle may be maintained at an acidic pH. Exposure of cells to acridine orange, a fluorescent amine that accumulates in acidic intracellular compartments, leads to its uptake and concentration within trichocysts. Thus intratrichocyst pH appears to be acidic in vivo and may serve as a regulatory or "safety" mechanism to inhibit premature expansion.     

Primary and secondary critical ischemia times of myocutaneous flaps

The primary critical ischemia time of the latissimus dorsi myocutaneous flap model was determined in the pig. Latissimus dorsi flaps were subjected to a primary ischemic insult of 2 hours (mimicking the ischemic event of free-tissue transfer). Following 12 hours of normal flow, the flaps were subjected to a second ischemic insult ranging from 0 to 12 hours. The secondary critical ischemia time (11.3 hours) was found to be statistically comparable to the primary critical ischemia time (9.1 hours). Questions are raised concerning the mechanism of action of this phenomenon and its clinical relevance.     

HLA-JY328: Mapping studies and expression of a polymorphic HLA class I gene

The JY328 clone was identified in a human genomic library using cDNA corresponding to mRNA for HLA-B7 as a probe. The L/328 cell line was established by cotransformation of mouse Ltk^– cells with the herpes thymidine kinase gene and clone JY328. On Northern blots, RNA from,L/328 strongly hybridized to an HLA class I probe, and an antigen was recognized by an anti-HLA class I framework antibody on the cell surface. A DNA probe corresponding to a segment of intron 7 was developed by comparing the nucleotide sequence of clone JY328 with that of other HLA class I-type genes. Using the radiolabeled probe to screen Southern blots of DNA from families with siblings exhibiting intra-HLA recombinations, a restriction fragment length polymorphism was revealed —a 1.4 kb BstE II band not present in all individuals. A corresponding fragment was apparent in the base sequence of clone JY328. The occurrence of this band on Southern blots established that JY328 maps distinct from and centromeric to the HLA-C locus and near to the HLA-B locus. Antibody absorption studies and cytotoxicity tests indicated that the JY328 gene product was not an HLA-B antigen but that it did specifically absorb CW7-specific antibody. In sum, these results suggest a novel, polymorphic HLA class I gene which expresses a product serologically similar to HLA-Cw7 but which does not map within the corresponding locus.