Sources of fungal linamarases

Forty-four strains of Aspergillus, Penicillium, Fusarium, Trichoderma and Rhizopus were grown on a liquid medium containing glucose and cassava-root extract. All of the Aspergillus and Fusarium strains, eight out of 10 Penicillium strains and three of seven Trichoderma strains showed linamarase activity. No such activity was detected in any Rhizopus strain. The crude enzyme preparation from F. oxysporum had the highest affinity for linamarin whereas that from A. nidulans was the most heat-stable.The authors are with the Department of Botany, National University of Singapore. Kent Ridge. Singapore 0511     

Pain due to lesions of central nervous system removed by sympathetic block.

Eight patients were studied in whom a lesion within the central nervous system caused constant pain and hyperpathia. Blockade of the sympathetic supply to the periphery was carried out in each patient by stellate ganglion block or intravenous infusion of guanethidine 15 mg in 30 ml saline into a limb on the affected side. On almost every occasion the pain and hypersensitivity were reduced, sometimes completely. Thus chronic pain and hyperpathia arising from a lesion in the central nervous system may be abolished by blocking the sympathetic supply to the periphery; this effect may be achieved when not all the peripheral nerves of the affected region have had their sympathetic nerve supply blocked. Such blockade may be worth repeating in the hope of achieving lasting relief of the intractable pain.     

Non-invasive genotyping of Helicobacter pylori cagA, vacA, and hopQ from asymptomatic children

Background: Helicobacter pylori infection is usually acquired in childhood, but little is known about its natural history in asymptomatic children, primarily due to the paucity of non‐invasive diagnostic methods. H. pylori strains harboring cagA and specific alleles of hopQ and vacA are associated with increased risk for gastric cancer. Many studies of H. pylori virulence markers in children have the bias that symptomatic subjects are selected for endoscopy, and these children may harbor the most virulent strains. Our aim is to genotype cagA, hopQ, and vacA alleles in stool DNA samples of healthy Colombian children residing in an area with high incidence of gastric cancer, to avoid selection bias resulting from endoscopy. Methods: H. pylori status of 86 asymptomatic children was assessed by ¹³C‐urea breath test (UBT) and PCR. H. pylori 16S rRNA, cagA, hopQ, and vacA genes were amplified from stool DNA samples and sequenced. Results: UBT was positive in 69 (80.2%) of 86 children; in stool DNA analysis, 78.3% were positive by 16S rRNA PCR. cagA, vacA, and hopQ were detected in 66.1%, 84.6%, and 72.3% of stool DNA samples from 16S rRNA‐positive children. Of the children’s DNA samples, which revealed vacA and hopQ alleles, 91.7% showed vacA s1 and 73.7% showed type I hopQ. Type I hopQ alleles were associated with cagA positivity and vacA s1 genotypes (p < 0.0001). Conclusions: Using stool DNA samples, virulence markers of H. pylori were successfully genotyped in a high percentage of the asymptomatic infected children, revealing a high prevalence of genotypes associated with virulence. Type I hopQ alleles were associated with the presence of cagA and the vacA s1 genotype.     

New records of the Japanese seahorse Hippocampus mohnikei in Southeast Asia lead to updates in range,habitat and threats

New records of the Japanese seahorse Hippocampus mohnikei from Cambodia, Malaysia, Thailand and Vietnam, along with recently published studies from India and Singapore, have greatly expanded the known range of H. mohnikei within Southeast Asia. These new records reveal novel habitat preferences and threats to H. mohnikei in the region. Although the global conservation status of H. mohnikei is classified as Data Deficient according to the IUCN Red List of Threatened Species, new sightings indicate that this species is found in similar habitats and faces similar threats as other Hippocampus species that are considered Vulnerable.     

The GTP-binding protein YqeH participates in biogenesis of the 30S ribosome subunit in Bacillus subtilis

Bacterial genome sequencing has revealed a novel family of P-loop GTPases that are often essential for growth. Accumulating evidence suggests that these proteins are involved in biogenesis of the 30S or 50S ribosomal subunits. YqeH is a member of this Obg/Era GTPase family, with its function remains to be uncovered. Here, we present results showing that YqeH is involved in the 30S subunit biogenesis in Bacillus subtilis. We observed a reduction in the 70S ribosome and accumulation of the free 50S subunit in YqeH-depleted cells. Interestingly, no free 30S subunit accumulation was evident. Consistent with the theory that YqeH is involved in 30S subunit biogenesis, a precursor of 16S rRNA and its degradation products were detected. Additionally, the reduction of free 30S subunit was not observed in Era-depleted cells. YqeH overexpression did not compensate for growth defects in mutants devoid of Era and vice versa. Moreover, in vitro GTPase analyses showed that YqeH possessed high intrinsic GTPase activity. In contrast, Era showed slow GTPase activity, which was enhanced by the 30S ribosomal subunit. Our findings strongly suggest that YqeH and Era function at distinct checkpoints during 30S subunit assembly. B. subtilis yqeH is classified as an essential gene due to the inability of the IPTG-dependent P(spac)-yqeH mutant to grow on LB or PAB agar plates in the absence of IPTG. However, in our experiments, the P(spac)-yqeH mutant grew in PAB liquid medium without IPTG supplementation, albeit at an impaired rate. This finding raises the interesting possibility that YqeH participates in assembly of the 30S ribosomal subunit as well as other cellular functions essential for growth on solid media.     

Quasielastic light scattering from solutions of filamentous viruses. II. Comparison between theories and experiments

We have compared four theoretical effects of rodlike macromolecules with the fast components, i.e., components other than translational diffusion, of our experimental data, which are presented as amplitude autocorrelation functions of electric field scattered from dilute solutions of monodisperse rodlike viruses with lengths from 3300 Å for tobacco mosaic virus to 20,000 Å for Pf1. The four effects are (1) the optic anisotropy treated by Aragón and Pecora, (2) coupled translational–rotational diffusion due to anisotropy in translational mobility recently reformulated by Gierke, (3) anisotropic rotational diffusion with respect to the direction of translational displacement first discussed by Berne and Pecora, and (4) the bending mode of a rod by Fujime and Maruyama. We show that both the first and second effects are required to explain the enhancement of amplitude of the translational diffusion at the expense of fast components. The experimental decay rates of the fast component exceed that of the rotational diffusions. In order to explain the excessive decay rate in the fast component, we need to include a minute amount (～1%) of bending mode of rodlike viruses, especially in longer viruses such as M13 and Pf1.     

An in vitro method for rapid regeneration of a monopodial orchid hybrid Aranda Deborah using thin section culture

A thin section culture system for rapid regeneration of the monopodial orchid hybrid Aranda Deborah has been developed. Thin sections (0.6–0.7mm thick) obtained by transverse sectioning of a single shoot tip (6–7mm), when cultured in Vacin and Went medium enriched with coconut water (20% v/v), produced an average 13.6 protocorm-like bodies (PLB) after 45 days, compared to 2.7 PLB formed by a single 6–7 mm long shoot tip under same culture condition. Addition of -naphthaleneacetic acid to Vacin and Went medium enriched with coconut water further increased PLB production by thin sections. PLB developed into plantlets on solid Vacin and Went medium containing 10% (v/v) coconut water and 0.5 g l^–1 activated charcoal. With this procedure, more than 80,000 plantlets could be produced from thin sections obtained from a single shoot tip in a year as compared to nearly 11,000 plantlets produced by the conventional shoot tip method.Abbreviations BA 6-benzyladenine - CD callus development - CW coconut water - KC Knudson C medium - MS Murashige and Skoog medium - NAA -naphthaleneacetic acid - PLB protocorm-like body - TS thin section - VW Vacin and Went medium     

Preparation of acetylenic sugar derivatives by way of 2-(N-nitroso)acetamido sugars

N-Nitrosation with dinitrogen tetraoxide was used to convert 2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-α-D-glucopyranose (1) and 2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-β-D-galactopyranose (4) in high yield into the N-nitroso derivatives 2 and 5, respectively. Similarly, 3-acetamido-1,2,4,6-tetra-O-acetyl-3-deoxy-β-D-glucopyranose (12) and methyl 2-acetamido-3,4,5,6-tetra-O-acetyl-2-deoxy-D-gluconate (15) gave their respective, crystalline N-nitroso derivatives 13 and 16. Various other 2-acetamido sugar derivatives were likewise nitrosated. In ethereal solution, compounds 2 and 16 reacted with potassium hydroxide in isopropyl alcohol to give the C₅ acetylene, 1,2-dideoxy-D-erythro-pent-1-ynitol, isolated as the known triacetate 3. By the same procedure, the galacto derivative 5 was converted in high yield into the 3-epimeric C₅ acetylene, 1,2-dideoxy-D-threo-pent-1-ynitol, isolated as its triacetate 6 and characterized by conversion into the known, crystalline 1,2-dideoxy-3-O-(3,5-dinitrobenzoyl)-4,5-O-isopropylidene-D-threo-pent-1-ynitol (7).     

Discrete changes in brain calcium with morphine analgesia, tolerance-dependence, and abstinence

Subcellular localization of ⁴⁵Ca⁺⁺ in brain was determined after intracerebroventricular injection of the isotope in mice. Acute morphine injection selectively depleted ⁴⁵Ca⁺⁺ from synaptic vesicles while chronic morphine treatment increased the ⁴⁵Ca⁺⁺ in vesicular fractions. The elevated vesicular ⁴⁵Ca⁺⁺ found in tolerant-dependent animals rapidly declined during naloxone precipitated abstinence. These effects of morphine on brain Ca⁺⁺ localization are discussed in terms of their possible relationship to neurotransmitter release and tolerance and dependence development.