The genome of Neisseria gonorrhoeae retains the remnants of a two-component regulatory system that once controlled piliation

An intact activator-binding site upstream of the sigma(54) promoter of the pilin-encoding pilE gene of Neisseria gonorrhoeae suggests gonococci produce a protein capable of binding this sequence. We cloned a chimeric gene, rsp, that has sequence similarity to both the pilS and pilR genes of Pseudomonas aeruginosa encoding a two-component regulatory system that controls piliation. This gene is transcribed in N. gonorrhoeae and indirect evidence suggests that Rsp binds to the activator-binding site of the pilE gene. Despite this, mutation of rsp has no effect on piliation in N. gonorrhoeae, suggesting that the remnants of this regulatory system have persisted in the genome, despite the loss of its original function.     

Spermiogenesis in the brush-tailed possum,Trichosurus vulpecula (Marsupialia)

Summary Acrosome development in the Australian Brush-tailed possum, Trichosurus vulpecula, displays a number of extraordinary features. This is particularly evident in the later stages of spermiogenesis, when the area of the nuclear surface bounded by the nuclear ring, and covered by the acrosome, is reduced considerable. As a result, the acrosomal material becomes located over its definitive position on the anterior third of the dorsal nuclear surface; in this process it is thrown into a series of folds, and a wide subacrosomal space is formed.Further changes around the time of spermiation result in the release of a spermatozoon in which a thin layer of acrosomal material is closely applied to the nucleus over the area of the definitive location of the acrosome, whilst its margins are greatly extended and project freely away from the nucleus. The latter feature does not appear to have been reported for the sperm of other mammals.The authors would like to thank Dr. D.J.H. Cockayne, Director of the Electron Microscope Unit, University of Sydney, for the generous provision of transmission electron microscope facilities, and Dr. M.R. Dickson, Electron Microscopist in charge, Biomedical Electron Microscope Unit, University of New South Wales, for the use of other facilities. Also, thanks are due to Miss Robin Arnold and Mrs. Eva Vassak of the Department of Histology and Embryology, University of Sydney, for their expert assistance. The assistance of the N.S.W. National Parks and Wildlife Service in the provision of permits to work on these native mammals is acknowledged     

Some observations on the growth and length: weight relationship of the South Georgia cod Notothenia rossii marmorata Fischer during the first four years of life

The paper describes the length: weight relationship and the results of back-calculating fish length from scale length for Notothenia rossii marmorata Fischer, collected in April 1961 from Leith Harbour, South Georgia.     

Comparison of complete nuclear receptor sets from the human, Caenorhabditis elegans and Drosophila genomes

Background

The availability of complete genome sequences enables all the members of a gene family to be identified without limitations imposed by temporal, spatial or quantitative aspects of mRNA expression. Using the nearly completed human genome sequence, we combined in silico and experimental approaches to define the complete human nuclear receptor (NR) set. This information was used to carry out a comparative genomic study of the NR superfamily.

Results

Our analysis of the human genome identified two novel NR sequences. Both these contained stop codons within the coding regions, indicating that both are pseudogenes. One (HNF4 γ-related) contained no introns and expressed no detectable mRNA, whereas the other (FXR-related) produced mRNA at relatively high levels in testis. If translated, the latter is predicted to encode a short, non-functional protein. Our analysis indicates that there are fewer than 50 functional human NRs, dramatically fewer than in Caenorhabditis elegans and about twice as many as in Drosophila. Using the complete human NR set we made comparisons with the NR sets of C. elegans and Drosophila. Searches for the >200 NRs unique to C. elegans revealed no human homologs. The comparative analysis also revealed a Drosophila member of NR subfamily NR3, confirming an ancient metazoan origin for this subfamily.

Conclusions

This work provides the basis for new insights into the evolution and functional relationships of NR superfamily members.     

The tandem CCCH zinc finger protein tristetraprolin and its relevance to cytokine mRNA turnover and arthritis

Tristetraprolin (TTP) is the best-studied member of a small family of three proteins in humans that is characterized by a tandem CCCH zinc finger (TZF) domain with highly conserved sequences and spacing. Although initially discovered as a gene that could be induced rapidly and transiently by the stimulation of fibroblasts with growth factors and mitogens, it is now known that TTP can bind to AU-rich elements in mRNA, leading to the removal of the poly(A) tail from that mRNA and increased rates of mRNA turnover. This activity was discovered after TTP-deficient mice were created and found to have a systemic inflammatory syndrome with severe polyarticular arthritis and autoimmunity, as well as medullary and extramedullary myeloid hyperplasia. The syndrome seemed to be due predominantly to excess circulating tumor necrosis factor-α (TNF-α), resulting from the increased stability of the TNF-α mRNA and subsequent higher rates of secretion of the cytokine. The myeloid hyperplasia might be due in part to increased stability of granulocyte–macrophage colony-stimulating factor (GM-CSF). This review highlights briefly the characteristics of the TTP-deficiency syndrome in mice and its possible genetic modifiers, as well as recent data on the characteristics of the TTP-binding site in the TNF-α and GM-CSF mRNAs. Recent structural data on the characteristics of the complex between RNA and one of the TTP-related proteins are reviewed, and used to model the TTP-RNA binding complex. We review the current knowledge of TTP sequence variants in humans and discuss the possible contributions of the TTP-related proteins in mouse physiology and in human monocytes. The TTP pathway of TNF-α and GM-CSF mRNA degradation is a possible novel target for anti-TNF-α therapies for rheumatoid arthritis, and also for other conditions proven to respond to anti-TNF-α therapy.     

Evidence for consumer regulation of biofilm–nutrient interactions among hardwater streams (Pennsylvania, USA)

Experimental studies evaluating the simultaneous effects of consumers, nutrients, and other biotic/abiotic factors on intact, natural food webs are rare, particularly among ecosystems of varying trophic conditions. We conducted a series of in situ studies that used nutrient-diffusing substrata with nitrogen (N) and phosphorus (P) concentrations in a full factorial design in three temperate, limestone streams in Pennsylvania across a trophic gradient (mesotrophic, eutrophic, and hypereutrophic streams). We assessed differences in algal and macroinvertebrate biomass, taxonomic composition, and functional groups relative to amended nutrients across the trophic gradient; as such, these results facilitated predictions about regulators of food web structure. All factors varied significantly among the streams (e.g., algal biomass P = 0.005, macroinvertebrate biomass P < 0.001, algal diversity P = 0.006, macroinvertebrate diversity P < 0.001, algal group P < 0.001, macroinvertebrate guilds P < 0.001); the streams, however, did not exhibit simple responses to nutrient amendment. Algal and macroinvertebrate biomass and diversity responded greatest in the mesotrophic stream while grazing seemed to be a strong factor preventing algal nutrient response in the eutrophic and hypereutrophic streams. Brillouin’s Evenness Index was most influenced by nutrient amendment (nutrient effect on algae and macroinvertebrates P = 0.021). As such, we concluded that biomass and diversity were mediated by complexity within intermediate trophic levels.     

Changes of skeletal muscle adiponectin content in diet-induced insulin resistant rats

The current study examined the relationship between skeletal muscle levels of adiponectin and parameters of insulin sensitivity. A high fat/sucrose diet (HFD) for 20 weeks resulted in significant increases in body weight, serum insulin, triglycerides (TG), and free fatty acids (FFA) (all p < 0.01). Interestingly, this diet leads to a slight increase in serum adiponectin, but significant decreases in gastrocnemius muscle and white adipose adiponectin (all p < 0.05). HFD for 4 weeks also resulted in a significant decrease in muscle adiponectin, which correlated with serum insulin, TG, and FFA (all p < 0.05). Treatment of the 4-week HFD rats with a PPARgamma agonist GI262570 ameliorated the diet-induced hyperinsulinemia and dyslipidemia, and effectively restored muscle adiponectin (all p < 0.05). This study demonstrated that HFD-induced hyperinsulinemia and dyslipidemia appeared without changes in serum adiponectin, but were associated with decreased tissue adiponectin. This provides the first evidence for a connection between tissue adiponectin and diet-induced hyperinsulinemia and dyslipidemia.     

Arachidonic acid release in renal proximal tubule cell injuries and death

Arachidonic acid release and the effect of phospholipase inhibitors on various types of cell injuries and death to rabbit renal proximal tubule suspensions were determined. Proximal tubules were exposed to the mitochondrial inhibitor antimycin A (0.1 μM), the protonophore carbonyl cyanide ρ-trifluoromethoxypheitylhydrazone (1 μM FCCP), the oxidant tertbutyl hydroperoxide (0.5 mM TBHP), or the calcium ionophore ionomycin (5 μM) in the absence or presence of the putative phospholipase inhibitors dibucaine, mepacrine, chlorpromazine, or U-26384. The phospholipase inhibitors had no effect on the proximal tubule lactate dehydrogenase (LDH) release (a marker of cell death) produced by FCCP, antimycin A, or ionomycin after 1,2, or 2 hours of exposure, respectively. Only dibucaine and mepacrine decreased LDH release in TBHP-treated proximal tubules without decreasing TBHP-induced lipid peroxidation. Antimycin A and ionomycin did not release arachidonic acid from proximal tubules prelabeled with [1-¹⁴C] arachidonic acid. In contrast, TBHP released arachidonic acid from proximal tubules prior to the onset of cell death, and dibucaine and mepacrine decreased the TBHP-induced release. Thus, phospholipase inhibitors were cytoprotective in those injuries that produced arachidonic acid release. These results suggest that arachidonic acid release and phospholipase A₂ activation play a contributing role in oxidant-induced renal proximal tubule cell injury and death but not in mitochondrial inhibitor- or calcium ionophore-induced proximal tubule cell injury and death.     

Improved methods for detecting enteric viruses in oysters.

New and improved methods for concentrating enteroviruses, reoviruses, and adenoviruses from oysters have been developed and evaluated. Viruses are efficiently adsorbed to homogenized oyster meat by adjusting the homogenate to pH 5.0 and a conductivity of less than or equal to 2,000 mg of NaCl per liter. After low-speed centrifugation, the virus-free supernatant is discarded and the viruses are eluted from the sedimented oyster solids with pH 7.5 glycine-NaCl having a conductivity of 8,000 mg of NaCl per liter. The oyster solids are removed by low-speed centrifugation and filtration, and the viruses in the filtered supernatant are concentrated to a small volume by either ultrafiltration or acid precipitation at pH 4.5. The concentrate is treated with antibiotics and inoculated into cell cultures for virus isolation and quantitation. When these methods were tested with oysters experimentally contaminated with polioviruses, reoviruses, and adenoviruses, recovery efficiencies averaged about 46%. With the exception of virus assay and quantitation, these methods are simple and inexpensive enough to be done in typical shellfish microbiology laboratories.     

Membranous Inclusions of Pseudomonas aeruginosa

Electron microscopy of sectioned cells of Pseudomonas aeruginosa treated with a double fixative in N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid revealed a number of membranous inclusions varying in size and morphology. One round, electron transparent form was frequently observed which did not routinely appear to be attached to the cytoplasmic membrane and varied in size from 120 to 300 nm in diameter. However, in one case, several tubular structures between an inclusion and the cytoplasmic membrane was observed. On rare occasions, a large and unusual multilayered inclusion consisting of three thick and distinct layers was also encountered. In addition, two small mesosomal structures were singly observed in cells and were situated proximal to the cytoplasmic membrane. One type appeared to consist of a single thin membrane, whereas the other type consisted of a delicate, multilayered structure.