Nuclear migration advances in fungi

Nuclear migration encompasses three areas: separation of daughter nuclei during mitosis, congress of parental nuclei before they fuse during fertilization, and positioning of nuclei in interphase cells. This review deals primarily with interphase nuclear migration, which is crucial for events as disparate as vertebrate embryonic development and growth of fungal mycelia. Mutants of Aspergillus nidulans, Neurospora crassa and Saccharomyces cerevisiae have been particularly informative, and a detailed molecular analysis of this process is now well under way.     

XM-6: A new gel-forming bacterial polysaccharide

A new extracellular microbial polysaccharide, XM-6, has been isolated from cultures of an Enterobacter species and shows unusual gelation properties of potential technological significance. The polysaccharide contains d-glucose, l-fucose and d-glucuronate in the approximate molar ratio 3:1:1. No significant amounts of acetate or pyruvate were detected. d-Glucuronate and some d-glucose are destroyed on periodate oxidation, but l-fucose and some d-glucose may be recovered intact, indicating the presence of some 1,3 linkages in the primary structure. The major oligosaccharide isolated from autohydrolysates was an aldobiuronic acid containing equal amounts of d-glucose and l-fucose.Thermally-reversible gels are formed on addition of salt to solutions of the polysaccharide. A preliminary investigation of the mechanism of gelation by optical rotation, circular dichroism, high resolution n.m.r. and mechanical spectroscopy suggests interchain association through conformationally ordered ‘junction zones’, with specific incorporation of site-bound cations within the ordered structures. In the sol state the polysaccharide shows the shear-rate and temperature dependence of viscosity typical of a disordered (‘random coil’) polymer solution. Divalent cations are, in general, more effective than monovalent cations in promoting gelation of XM-6, while trivalent cations normally cause precipitation. Within Groups I and II, optimum gelation is achieved with Na⁺ and Ca²⁺ (ionic radius ? 0·1 nm), with larger and smaller ions becoming progressively less effective. Both gel strength and melting temperature increase with increasing salt concentration.XM-6 forms gels of reasonable strength at unusually low concentrations of the polysaccharide. For example, gels comparable to those required for normal industrial or food applications may be obtained using 0·3% w/v XM-6 and 1% w/v NaCl. Gel strength increases with increasing polymer concentration but there is no systematic variation in melting point. The sol-gel transition of XM-6 is unusually sharp and, by suitable adjustment of salt concentration, can be made to occur just below body temperature (e.g. 30–35°C), with obvious implications for biomedical or food applications.     

Cold shock injury in Tetrahymena pyriformis

The ciliated protozoan Tetrahymena pyriformis has been used to study the biochemistry of cellular injury induced by rapid cooling (cold shock). Cellular viability was found to depend on the time and temperature of cold exposure, and the rate of cooling. During cooling to −7.5 °C, in the absence of ice, an optimal rate of cooling of 2.5 °C min⁻¹ was observed; at both faster and slower cooling the recovery decreased. Following acclimation at a reduced temprature (10 °C) the viability following rapid cooling was significantly different from that of cultures maintained at 20 °C. Analysis of the phospholipid fatty acids from cells grown at 10 °C demonstrated that, at the reduced temperature, there was an increase in the average degree of fatty acyl unsaturation. Cold-shock injury in Tetrahymena is associated with membrane thermotropic events which are determined by temperature per se, whereas viability is a function of the rate of cooling. A hypothesis of injury is presented in which the presence of gel-phase lipid within the membrane is not the critical event, but it is the pattern of nucleation within the membrane which ultimately determines the extent of cellular injury.     

Production of Solvents by Clostridium acetobutylicum Cultures Maintained at Neutral pH

The formation of acetone and n-butanol by Clostridium acetobutylicum NCIB 8052 (ATCC 824) was monitored in batch culture at 35°C in a glucose (2% [wt/vol]) minimal medium maintained throughout at either pH 5.0 or 7.0. At pH 5, good solvent production was obtained in the unsupplemented medium, although addition of acetate plus butyrate (10 mM each) caused solvent production to be initiated at a lower biomass concentration. At pH 7, although a purely acidogenic fermentation was maintained in the unsupplemented medium, low concentrations of acetone and n-butanol were produced when the glucose content of the medium was increased (to 4% [wt/vol]). Substantial solvent concentrations were, however, obtained at pH 7 in the 2% glucose medium supplemented with high concentrations of acetate plus butyrate (100 mM each, supplied as their potassium salts). Thus, C. acetobutylicum NCIB 8052, like C. beijerinckii VPI 13436, is able to produce solvents at neutral pH, although good yields are obtained only when adequately high concentrations of acetate and butyrate are supplied. Supplementation of the glucose minimal medium with propionate (20 mM) at pH 5 led to the production of some n-propanol as well as acetone and n-butanol; the final culture medium was virtually acid free. At pH 7, supplementation with propionate (150 mM) again led to the formation of n-propanol but also provoked production of some acetone and n-butanol, although in considerably smaller amounts than were obtained when the same basal medium had been fortified with acetate and butyrate at pH 7.     

Cyanophycin granule polypeptide protease in a unicellular cyanobacterium

An assay was developed to measure the proteolysis of cyanophycin granule polypeptide in crude extracts of a unicellular cyanobacterium. The substrate was radioactively labeled cyanophycin granule polypeptide formed by an unicellular cyanobacterium grown in the presence of chloramphenicol. Substrate polypeptide displayed identical chemical properties with polypeptide isolated from non-chloramphenicol-treated cells. Solubilization of radioactivity as arginine indicated hydrolysis of polypeptide. Radioactively labeled aspartate and arginine from hydrolyzed polypeptide was related to nmol amino acid using a combination of paper chromatography, liquid scintillation analysis, and ninhydrin quantitation. Protease activity was found in extracts of nitrogen-limited cells harvested 16–24 h after a nitrogen source was added back. Optimal pH for protease activity was 8.0 and optimum temperature was 35°C. Protease activity in crude extracts followed Michaelis-Menten kinetics with a V _max of 92 nmol arginine per 15 min/mg protein and a K _m of 2.1×10³ nmol arginine. Protease activity was inhibited by arginine and by high concentrations of aspartate.     

Induction of ovulation with pulsatile luteinising hormone releasing hormone.

Ovulation was successfully induced with luteinising hormone releasing hormone in 28 women with hypothalamic amenorrhoea who had failed to respond to treatment with clomiphene. Luteinising hormone releasing hormone was administered in a pulsatile manner with miniaturised automatic infusion systems. The rate of ovarian follicular maturation, as monitored by serial pelvic ultrasonography, was similar to that observed in spontaneous cycles. Endocrine assessment by serial measurement of gonadotrophin, oestradiol, and progesterone concentrations showed hormone concentrations to be within the normal range. Intravenous treatment was required in only two patients, the remainder responding satisfactorily to subcutaneous infusion. All patients conceived within six cycles of treatment, and only one multiple pregnancy occurred.     

Is there a place in the United Kingdom for intensive antacid treatment for chronic peptic ulceration?

Sixty nine patients with chronic duodenal or juxtapyloric ulceration were studied in a prospective double blind randomised trial to compare the efficacy of antacid and placebo at high (30 ml seven times daily) and low (10 ml as required) doses. After four weeks ulcers had healed in 12 out of 18 patients (67%) receiving "low dose" antacid compared with in six out of 17 patients (35%) receiving low dose placebo; ulcers had also healed in six out of 19 patients (32%) receiving "high dose" antacid compared with in two out of 15 patients (13%) receiving high dose placebo. Overall, the effect of antacid was superior to that of placebo in healing ulcers (p less than 0.05) and the effect of low dose treatment was superior to that of high dose treatment (p less than 0.01). There were no significant differences between antacid and placebo at eight weeks. Antacid was better than placebo in relieving pain, but the difference was not significant. Poor compliance and high incidence of diarrhoea made high dose antacid an impractical treatment. Low dose antacid was associated with a significantly better rate of healing than high dose antacid and was far better tolerated. This low dosage of antacid should be considered to be an active treatment in trials of ulcer healing.     

Renal vascular anatomy of the toad (Bufo marinus)

The renal vasculature of the toad, Bufo marinus, was studied mainly by means of scanning electron microscopy of vascular corrosion casts. All arterial branches terminated in a glomerulus. Each glomerulus was supplied by only one afferent arteriole. No shunts between afferent and efferent arterioles were observed. The glomerular channels appeared to be permanent capillaries. No evidence supporting the theory of freely shifting glomerular blood channels was found. Efferent arterioles radiated out towards the dorsal surface of the kidney where they connected with peritubular vessels. The renal portal veins produced an anastomosing plexus on the dorsal surface of the kidney, giving rise to the peritubular vessels. Peritubular vessels ran radially toward the ventral surface of the kidney, where they formed the roots of the renal veins. Attention is drawn to the possibility of hairpin countercurrent exchange between the capillary-like efferent arterioles and the peritubular vessels in the dorsal kidney.     

Purification and structural analysis of the fourth component of human complement.

The fourth component of human complement (C4) has been purified in 20% yield from fresh plasma using as starting material the 5-12% poly(ethylene glycol) precipitate which had been depleted of plasminogen by an affinity adsorbent. Sequential ion-exchange chromatography on diethylaminoethylcellulose, QAE-Sephadex, and DEAE-Bio-Gel A resulted in C4 homogeneous by immunological criteria and by polyacrylamide gel electrophoresis, the last chromatographic step achieving separation of native from inactivated C4. Reduction with 20 mM dithiothreitol for 2 h at 37 degrees C in 0.25 M 2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride, pH 8.6, effected cleavage of the interchain disulfide bonds. A three-chain structure for C4 was confirmed, and molecular weight estimates of 93 000 +/- 9300, 75 000 +/- 7500, and 30 000 +/- 3000 determined for the alpha, beta, and gamma chains, respectively. The effects of known inactivators of C4 upon the chains of C4 were investigated, confirming that the inactivations by C1s and trypsin were accompanied by the fragmentation of the alpha chain. Inactivation of C4 by hydrazine, on the other hand, produced no detectable change in chain size. Separation of the chains was accomplished by gel filtration in the presence of 1 M acetic acid. Amino acid compositions of native C4 and the constitutive chains have been performed, and N-terminal sequences of the latter established by automated Edman degradation.