Genome sequence of the emerging pathogen Aeromonas caviae

Aeromonas caviae is a Gram-negative, motile and rod-shaped facultative anaerobe that is increasingly being recognized as a cause of diarrhea in children. Here we present the first genome sequence of an A. caviae strain that was isolated as the sole pathogen from a child with profuse diarrhea.     

Structural and biological properties of the Drosophila insulin-like peptide 5 show evolutionary conservation

We report the crystal structure of two variants of Drosophila melanogaster insulin-like peptide 5 (DILP5) at a resolution of 1.85 Å. DILP5 shares the basic fold of the insulin peptide family (T conformation) but with a disordered B-chain C terminus. DILP5 dimerizes in the crystal and in solution. The dimer interface is not similar to that observed in vertebrates, i.e. through an anti-parallel β-sheet involving the B-chain C termini but, in contrast, is formed through an anti-parallel β-sheet involving the B-chain N termini. DILP5 binds to and activates the human insulin receptor and lowers blood glucose in rats. It also lowers trehalose levels in Drosophila. Reciprocally, human insulin binds to the Drosophila insulin receptor and induces negative cooperativity as in the human receptor. DILP5 also binds to insect insulin-binding proteins. These results show high evolutionary conservation of the insulin receptor binding properties despite divergent insulin dimerization mechanisms.     

Dependence of proteasome processing rate on substrate unfolding

Protein degradation by eukaryotic proteasomes is a multi-step process involving substrate recognition, ATP-dependent unfolding, translocation into the proteolytic core particle, and finally proteolysis. To date, most investigations of proteasome function have focused on the first and the last steps in this process. Here we examine the relationship between the stability of a folded protein domain and its degradation rate. Test proteins were targeted to the proteasome independently of ubiquitination by directly tethering them to the protease. Degradation kinetics were compared for test protein pairs whose stability was altered by either point mutation or ligand binding, but were otherwise identical. In both intact cells and in reactions using purified proteasomes and substrates, increased substrate stability led to an increase in substrate turnover time. The steady-state time for degradation ranged from ～5 min (dihydrofolate reductase) to 40 min (I27 domain of titin). ATP turnover was 110/min./proteasome, and was not markedly changed by substrate. Proteasomes engage tightly folded substrates in multiple iterative rounds of ATP hydrolysis, a process that can be rate-limiting for degradation.     

Toxicity of seven termiticides on the formosan and eastern subterranean termites

Using both topical application and substrate (sand) treatments the toxicities of seven new generation soil termiticides were evaluated to determine the LD50 and LC50 against two economically important subterranean termite species, eastern subterranean termite, Reticulitermes flavipes (Kollar), and Formosan subterranean termite, Coptotermes formosanus Shiraki. The lethal dose toxicity (LD50) rankings for R. flavipes from highest to lowest were: fipronil > bifenthrin > chlorantraniliprole > cyantraniliprole > imidacloprid > chlorfenapyr > indoxacarb; the rankings for C. formosanus were fipronil > imidacloprid > chlorantraniliprole > cyanthraniliprole> bifenthrin > chlorfenapyr > indoxacarb. The respective lethal concentration toxicity (LC50) rankings were fipronil > bifenthrin > chlorfenapyr > indoxacarb > cyantraniliprole > chlorantraniliprole > imidacloprid for R. flavipes; and fipronil > chlorfenapyr > bifenthrin >imidacloprid > cyantraniliprole > chlorantraniliprole > indoxacarb for C. formosanus. The study provides an opportunity to directly compare toxicity, action speed, and bioavailability among this group of newer generation soil termiticides.     

In situ B cell-mediated immune responses and tubulointerstitial inflammation in human lupus nephritis

The most prevalent severe manifestation of systemic lupus erythematosus is nephritis, which is characterized by immune complex deposition, inflammation, and scarring in glomeruli and the tubulointerstitium. Numerous studies indicated that glomerulonephritis results from a systemic break in B cell tolerance, resulting in the local deposition of immune complexes containing Abs reactive with ubiquitous self-Ags. However, the pathogenesis of systemic lupus erythematosus tubulointerstitial disease is not known. In this article, we demonstrate that in more than half of a cohort of 68 lupus nephritis biopsies, the tubulointerstitial infiltrate was organized into well-circumscribed T:B cell aggregates or germinal centers (GCs) containing follicular dendritic cells. Sampling of the in situ-expressed Ig repertoire revealed that both histological patterns were associated with intrarenal B cell clonal expansion and ongoing somatic hypermutation. However, in the GC histology, the proliferating cells were CD138(-)CD20(+) centroblasts, whereas they were CD138(+)CD20(low/-) plasmablasts in T:B aggregates. The presence of GCs or T:B aggregates was strongly associated with tubular basement membrane immune complexes. These data implicate tertiary lymphoid neogenesis in the pathogenesis of lupus tubulointerstitial inflammation.     

Tilt-pair analysis of images from a range of different specimens in single-particle electron cryomicroscopy

The comparison of a pair of electron microscope images recorded at different specimen tilt angles provides a powerful approach for evaluating the quality of images, image-processing procedures, or three-dimensional structures. Here, we analyze tilt-pair images recorded from a range of specimens with different symmetries and molecular masses and show how the analysis can produce valuable information not easily obtained otherwise. We show that the accuracy of orientation determination of individual single particles depends on molecular mass, as expected theoretically since the information in each particle image increases with molecular mass. The angular uncertainty is less than 1° for particles of high molecular mass (∼ 50 MDa), several degrees for particles in the range 1-5 MDa, and tens of degrees for particles below 1 MDa. Orientational uncertainty may be the major contributor to the effective temperature factor (B-factor) describing contrast loss and therefore the maximum resolution of a structure determination. We also made two unexpected observations. Single particles that are known to be flexible showed a wider spread in orientation accuracy, and the orientations of the largest particles examined changed by several degrees during typical low-dose exposures. Smaller particles presumably also reorient during the exposure; hence, specimen movement is a second major factor that limits resolution. Tilt pairs thus enable assessment of orientation accuracy, map quality, specimen motion, and conformational heterogeneity. A convincing tilt-pair parameter plot, where 60% of the particles show a single cluster around the expected tilt axis and tilt angle, provides confidence in a structure determined using electron cryomicroscopy.     

Relative humidity preference and survival of starved Formosan subterranean termites (Isoptera: Rhinotermitidae) at various temperature and relative humidity conditions

Foraging groups of Formosan subterranean termites, Coptotermes formosanus Shiraki were tested for their relative humidity (RH) preference in a humidity gradient arena in the laboratory at a constant temperature of 26°C. Five RH levels (9%, 33%, 53%, 75%, and 98%) were maintained in the test arena comprising of a series of closed containers by using dry silica gel, saturated salt solutions, or distilled water alone. Termites gradually aggregated to the highest RH chamber in the arena. After 1 h, a significantly greater percentage of termites (≈46%) aggregated to the highest RH chamber (98%) than to the lower RH chambers (≤75%). After 12 h, > 97% of the termites aggregated to the 98% RH chamber. In survival tests, where termites were exposed to 15 combinatorial treatments of five RH levels (9%, 33%, 53%, 75%, and 98%) and three temperatures (20°C, 28°C, and 36°C) for a week, the survival was significantly influenced by RH, temperature, and their interaction. A significantly higher mortality was observed on termites exposed to ≤75% RH chambers than to 98% RH chamber at the three temperatures and significantly lower survival was found at 36°C than at 28°C or 20°C. The combination of temperature and RH plays an important role in the survival of C. formosanus.     

Growth, ingestion rates and metabolic activity of walleye in lakes with and without lake herring

Growth efficiencies, ingestions rates and activity levels of walleye Sander vitreus were compared in lakes with and without lake herring Coregonus artedi . Yellow perch Perca flavescens were the main prey in lakes without lake herring. Walleye were sampled in September and October from 38 lakes in Ontario in 1998 and 1999, using multimesh monofilament gillnets. Ingestion rates were estimated from annual increments in somatic mercury and body mass, and the mercury content of yellow perch and lake herring. Walleye had higher growth efficiencies, and lower ingestion and activity rates in lakes with lake herring. Lake herring grow larger than yellow perch and therefore could provide more profitable prey for larger walleye. The results are consistent with optimal foraging theory that predicts that walleye feeding on optimal prey sizes should grow more efficiently, if the ratio of feeding benefit (energy) to cost (search and seizure) is a function of the ratio of predator and prey size.     

Adrenal lipid profiles of chemically sympathectomized normoxic and hypoxic neonatal rats.

Neonatal hypoxia is a common condition that elicits a coordinated endocrine response. In the neonatal rat, hypoxia induces an ACTH-independent increase in corticosterone which can be partially blocked by chemical sympathectomy. The present study sought to characterize the effects of sympathectomy on the adrenal lipid profile, since previous work suggested that augmented plasma corticosterone during hypoxia may be due to changes in adrenal lipid metabolism. Newborn rats were exposed to normoxia or hypoxia from birth to seven days of age, and guanethidine was used to produce the sympathectomy. Plasma epinephrine and norepinephrine were not significantly affected by hypoxia, while guanethidine decreased plasma norepinephrine in normoxic and hypoxic pups. Hypoxia alone increased the concentration of cholesterol esters in the adrenal gland; this increase was due to increases in cholesterol ester-associated oleic (18:1n9), docosahexaenoic (22:6n3), arachidonic (20:4n6), and adrenic (22:4n6) acids. Hypoxia also increased diglyceride-associated adrenic acid. Guanethidine treatment attenuated the hypoxia-induced increase in cholesterol ester-bound arachidonic and adrenic acids. Guanethidine also decreased saturated fatty acid concentrations and increased n3 fatty acid-enriched triglycerides. The results support the idea that the ACTH-independent corticosterone response to hypoxia in the neonatal rat is mediated by specific, sympathetically driven alterations in the adrenal lipid profile.