Structure and stereochemistry of brianolide, a new antiinflammatory diterpenoid from the Okinawan gorgonian Briareum sp.

Brianolide (1), a new antiinflammatory diterpenoid of the briarein class, possessing a beta substituent at C-12 (R), has been isolated from the Okinawan gorgonian Briareum sp. Its structure has been established from spectral data in conjunction with a single crystal X-ray analysis.     

In vitro inactivation of the neurotoxic action of β-bungarotoxin by the marine natural product,manoalide

Summary The irreversible neurotoxic action of -bungarotoxin (-BuTx) can be prevented by preincubation of the toxin with manoalide, a non-steroidal anti-inflammatory agent. Manoalide was also found to inactivate purified phospholipase A₂ and thus prevent hydrolysis of phosphatidylcholine. PLA₂ is a component found in several neurotoxic venoms and is also a rate limiting enzyme important in phospholipid metabolism and prostaglandin synthesis in man.This work is a result of research sponsored in part by NOAA-Department of Commerce under Sea Grant No. NA80AA-D-00120 and the California State Resources Agency R/MP-21. The US Government is authorized to produce and distribute reprints for governmental purposes. A preliminary report of aspects of this work appeared in Fedn Proc.42 (1983) 374. We also acknowledge the technical assistance of Dan Rohrer. Dr George Taborski's advice and assistance during the course of this work is gratefully acknowledged.J. C. de Freitas is a post-doctoral fellow supported by Fundacao de Amparo à Pesquisa do Estado de São Paulo (81/0263-9) and Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (201.171/82), Brazil.     

Human mitochondrial DNA deletions associated with mutations in the gene encoding Twinkle, a phage T7 gene 4-like protein localized in mitochondria.

The gene products involved in mammalian mitochondrial DNA (mtDNA) maintenance and organization remain largely unknown. We report here a novel mitochondrial protein, Twinkle, with structural similarity to phage T7 gene 4 primase/helicase and other hexameric ring helicases. Twinkle colocalizes with mtDNA in mitochondrial nucleoids. Screening of the gene encoding Twinkle in individuals with autosomal dominant progressive external ophthalmoplegia (adPEO), associated with multiple mtDNA deletions, identified 11 different coding-region mutations co-segregating with the disorder in 12 adPEO pedigrees of various ethnic origins. The mutations cluster in a region of the protein proposed to be involved in subunit interactions. The function of Twinkle is inferred to be critical for lifetime maintenance of human mtDNA integrity.     

Isolation and characterization of genomic and cDNA clones of human erythropoietin

The glycoprotein hormone erythropoietin regulates the level of oxygen in the blood by modulating the number of circulating erythrocytes, and is produced in the kidney or liver of adult and the liver of fetal or neonatal mammals. Neither the precise cell types that produce erythropoietin nor the mechanisms by which the same or different cells measure the circulating oxygen concentration and consequently regulate erythropoietin production are known. Cells responsive to erythropoietin have been identified in the adult bone marrow, fetal liver or adult spleen. In cultures of erythropoietic progenitors, erythropoietin stimulates proliferation and differentiation to more mature red blood cells. Detailed molecular studies have been hampered, however, by the impurity and heterogeneity of target cell populations and the difficulty of obtaining significant quantities of the purified hormone. Highly purified erythropoietin may be useful in the treatment of various forms of anaemia, particularly in chronic renal failure. Here we describe the cloning of the human erythropoietin gene and the expression of an erythropoietin cDNA clone in a transient mammalian expression system to yield a secreted product with biological activity.     

Relationships between Centaurea maculosa and indigenous plant assemblages

Ecological impacts of invasive plants include displacement of indigenous species and declines in species richness and diversity. The objective of this study was to characterize the functional relationship between plant community composition and Centaurea maculosa Lam. (spotted knapweed) within a Festuca idahoensis / Pseudoroegneria spicatum habitat type in Montana. Density, cover, and biomass of all species were collected along a gradient of spotted knapweed cover ranging from 0% to about 100%. Step-down regression was used to determine the relationship among C. maculosa , indigenous species, species richness, and Shannon-Weavers diversity index. Regressions showed that indigenous perennial grass cover, species richness, and species diversity were inversely related to C. maculosa cover. There was no relationship between C. maculosa and indigenous forbs. While this study does not imply a causal relationship, the literature suggests that C. maculosa displaces indigenous species and/or invades areas of reduced indigenous plant cover, low diversity, or low species richness. Knowing levels of indigenous perennial grass cover will help managers predict the outcome of weed management on rangelands that are vulnerable to weed infestation.     

Killing activity of neutrophils is mediated through activation of proteases by K+ flux

According to the hitherto accepted view, neutrophils kill ingested microorganisms by subjecting them to high concentrations of highly toxic reactive oxygen species (ROS) and bringing about myeloperoxidase-catalysed halogenation. We show here that this simple scheme, which for many years has served as a satisfactory working hypothesis, is inadequate. We find that mice made deficient in neutrophil-granule proteases but normal in respect of superoxide production and iodinating capacity, are unable to resist staphylococcal and candidal infections. We also show that activation provokes the influx of an enormous concentration of ROS into the endocytic vacuole. The resulting accumulation of anionic charge is compensated for by a surge of K+ ions that cross the membrane in a pH-dependent manner. The consequent rise in ionic strength engenders the release of cationic granule proteins, including elastase and cathepsin G, from the anionic sulphated proteoglycan matrix. We show that it is the proteases, thus activated, that are primarily responsible for the destruction of the bacteria.