ERp46 binds to AdipoR1, but not AdipoR2, and modulates adiponectin signalling

The pleiotropic effects of the insulin-sensitizing adipokine adiponectin are mediated, at least in part, by two seven-transmembrane domain receptors AdipoR1 and AdipoR2. Recent reports indicate a role for AdipoR-binding proteins, namely APPL1, RACK1 and CK2β, in proximal signal transduction events. Here we demonstrate that endoplasmic reticulum protein 46 (ERp46) interacts specifically with AdipoR1 and provide evidence that ERp46 modulates adiponectin signalling. Co-immunoprecipitation followed by mass spectrometry identified ERp46 as an AdipoR1-, but not AdipoR2-, interacting protein. Analysis of truncated constructs and GST-fusion proteins revealed the interaction was mediated by the cytoplasmic, N-terminal residues (1-70) of AdipoR1. Indirect immunofluorescence microscopy and subcellular fractionation studies demonstrated that ERp46 was present in the ER and the plasma membrane (PM). Transient knockdown of ERp46 increased the levels of AdipoR1, and AdipoR2, at the PM and this correlated with increased adiponectin-stimulated phosphorylation of AMPK. In contrast, adiponectin-stimulated phosphorylation of p38MAPK was reduced following ERp46 knockdown. Collectively these results establish ERp46 as the first AdipoR1-specific interacting protein and suggest a role for ERp46 in adiponectin receptor biology and adiponectin signalling.     

Association of beta-glucosidase with intact cells of Thermoactinomyces.

The location of the B-glucosidase activity in a whole culture broth of the thermophilic organism Thermoactinomyces has been studied. Little beta-glucosidase activity was found in the culture filtrate, while the culture solids contained the major part of the activity of the whole culture broth. The activity does not appear to be adsorbed to the culture solids; rather there is evidence that it is an intracellular soluble enzyme(s). The pH and temperature optima for a crude beta-glucosidase preparation were determined to be pH 6.5 and 50--55 degrees C. Enzyme activity studies indicate that the same enzyme(s) accounts for the beta-glucosidase and the cellobiase activities. The validity of using the filter paper activity of culture filtrates from Thermoactinomyces to predict the total saccharification of cellulosic materials to glucose is discussed.     

Effect of cellobiose, glucose, ethanol, and metal ions on the cellulase enzyme complex of Thermomonospora fusca

The saccharification of cellulosic substrates by cellulase from Thermomonospora fusca, strain YX, was influenced by the presence of various substances. Cellobiose was strongly inhibitory, reducing the activity against swollen cellulose to 25% at 5% concentration. Glucose had much less effect, reducing activity to 40% at 20% concentration. Ethanol was found to be only slightly inhibitory to the cellulase, reducing activity by about 15% at 6% concentration, but much more inhibitory to the cell-bound beta-glucosidase activity. Of the numerous metal ions examined, Ca(2+) and Co(2+) at 0.1mM-1.0mM concentration were found to be slightly activating under assay conditions, while 1.0mM Pb(2+) and Hg(2+) were the most inhibitory ions. The implications of these results for the design of commercial processes for ethanol production from cellulose are discussed.     

Bimodal distribution of constant frequencies in some hipposiderid bats (Mammalia: Hipposideridae)

Constant frequency echolocation signals of the Rhinolophidae and Hipposideridae have generally been found to occupy narrow bands, at least within single populations. Examination of three species of Hipposideridae from a cave in Kenya shows a wide spread of frequencies in each case. In Hipposideros commersoni the pulses occur in two bands at 56 and 66 kHz with a silent interval of 8 kHz. Single individuals in isolation use only one of these frequencies and this is correlated with their forearm length. It may be that two forms of subspecific rank were cohabiting, but it may be significant that the width of the silent band is just wide enough to prevent interference from band-spreading due to reflection from beating wings when several bats are flying together. In Triaenops afer and Hipposideros caffer there is a suggestion that the bats leave silent bands when in groups but use more varied individual frequencies when isolated. This impression might be due to sampling error and the position is little clarified by examining other populations. A group of Asellia tridens from Egypt occupied a single, fairly narrow band of frequencies. Further examination of these and other constant frequency species with a simple detector would be of interest in resolving the position.     

Variations in the middle ear of the Mammalia

The comparative morphology of mammalian middle ear structures has been studied by examining the histological sections of the ears of 144 specimens of mammals which had been collected, with a few exceptions, in two expeditions to the Caribbean area. Fifty-four species of six orders of mammals were represented. Sixteen structural features or relationships were specifically studied and the bearing of taxonomic groupings on the variability was analysed.
A taxonometric analysis using these middle ear features indicated that Microdipodops may lie nearer to Dipodomys than to Perognathus.
A table presenting the detailed results has been deposited with the British Museum (Natural History).     

Familial Site-specific Ovarian Cancer Is Linked to BRCA1 on 17q12-21

In a study of nine families with “site-specific” ovarian cancer (criterion: three or more cases of epithelial ovarian cancer and no cases of breast cancer diagnosed at age <50 years) we have obtained evidence of linkage to the breast-ovarian cancer susceptibility gene, BRCA1 on 17q12-21. If the risk of cancer in these families is assumed to be restricted to the ovary, the best estimate of the proportion of families linked to BRCA1 is .78 (95% confidence interval .32–1.0). If predisposition to both breast and ovarian cancer is assumed, the proportion linked is 1.0 (95% confidence interval .46–1.0). The linkage of familial site-specific ovarian cancer to BRCA1 indicates the possibility of predictive testing in such families; however, this is only appropriate in families where the evidence for linkage to BRCA1 is conclusive.     

Application of immunofluorescence to the establishment of an Encephalitozoon cuniculi-free rabbit colony.

Serologic screening of a rabbit breeding colony over a 9-month period showed that all 9-week-old rabbits with Encephalitozoon cuniculi infection were born of E cuniculi-infected does. This observation, obtained from studies on 395 young rabbits, suggested that transmission of infection is either transplacental or the result of close contact soon after birth. On this basis, 16 young healthy rabbits, seronegative to E cuniculi, were isolated and tested at 2-week intervals for antibodies to E cuniculi. In the first 2 months, seven rabbits showed indications of developing antibodies to E cuniculi and were immediately removed from the colony. The remaining rabbits along with their 52 offspring were tested for serum antibodies for a further 16 months and no rabbit became seropositive. Eight months after establishment of the colony, three does, one buck and six 12-week-old rabbits were killed. Macroscopic and extensive histologic and immunofluorescence examinations failed to reveal any evidence of infection with E cuniculi. These results showed that serological screening for E cuniculi infection by immunofluorescence is a simple yet adequate procedure for establishing a rabbit colony free of encephalitozoonosis.     

Cellulolytic Enzyme System of Thermoactinomyces sp. Grown on Microcrystalline Cellulose

The carboxymethyl-cellulase and Avicelase activities of Thermoactinomyces sp., strain YX, were produced simultaneously with cell growth. Throughout the growth phase these activities were primarily extracellular, with up to 50% adsorbed to residual cellulosic substrate at any one time. On the other hand, the β-glucosidase activity was associated with the culture solids throughout the entire fermentation and appears to be intracellular. Preparative isoelectric focusing of the culture filtrate, in the pH range 3 to 5, separated three major fractions with cellulolytic activities towards both carboxymethyl-cellulose and Avicel. Based on the carboxymethyl-cellulase and Avicelase activities of these separated fractions, it was not possible to discriminate between endo- and exoglucanases produced by Thermoactinomyces sp. Analytical isoelectric focusing of culture filtrates obtained throughout the growth phase of Thermoactinomyces indicated that all the extracellular cellulolytic enzymes are produced and released into the culture filtrate simultaneously, with no evidence of a sequential appearance of the various enzymes or isoenzymes.