Isolation and characterization of a meta-cleavage product in the degradation of quinaldic acid by Azotobacter sp.

Abstract From different samples of soil seventeen strains were isolated which grew aerobically in mineral salts medium with quinaldic acid as sole carbon source. Mutants were induced with N -methyl- N '-nitro- N -nitrosoguanidine. One mutant could be isolated which accumulated a yellow compound. The properties of this purified compound were those expected for 2-oxo-3-(4'-hydroxy-2'-oxo-3',4'-en-butyrate)-pyridine-6-carboxylic acid.     

Motor stereotypy and diversity in songs of mimic thrushes

The relationship between the motor and acoustic similarity of song was examined in brown thrashers (Toxostoma rufum) and grey catbirds (Dumetella carolinensis) (family Mimidae), which have very large song repertoires and sometimes mimic other species. Motor similarity was assessed by cross correlation of syringeal airflows and air sac pressures that accompany sound production. Although most syllables were sung only once in the song analyzed, some were repeated, either immediately forming a couplet, or after a period of intervening song, as a distant repetition. Both couplets and distant repetitions are produced by distinctive, stereotyped motor patterns. Their motor similarity does not decrease as the time interval between repetitions increases, suggesting that repeated syllables are stored in memory as fixed motor programs. The acoustic similarity between nonrepeated syllables, as indicated by correlation of their spectrograms, has a significant positive correlation with their motor similarity. This correlation is weak, however, suggesting that there is no simple linear relationship between motor action and acoustic output and that similar sounds may sometimes be produced by different motor mechanisms. When compared without regard to the sequence in which they are sung, syllables paired for maximum spectral similarity form a continuum with repeated syllables in terms of their acoustic and motor similarity. The prominence of couplets in the “syntax” of normal song is enhanced by the dissimilarity of successive nonrepeated syllables that make up the remainder of the song. © 1996 John Wiley & Sons, Inc.     

Synthesis and antitumour activity of derivatives of curdlan and lichenan branched at C-6.

Derivatives of curdlan and lichenan, linear (1----3)-beta-D- and (1----3/1----4)-beta-D-glucans, respectively, have been synthesised having alpha-L-arabinofuranosyl, alpha-L-rhamnosyl, beta-D-glucosyl, and beta-gentiobiosyl side chains attached at positions 6. These water-soluble derivatives, obtained by condensation of the 2,4- and 2,4-/2,3-di-O-phenylcarbamoyl derivatives of curdlan and lichenan, respectively, with appropriate ortho esters followed by saponification, were characterised by methylation analysis, g.p.c., and interaction with Congo Red. The curdlan derivatives and the lichenan derivative with few glucosyl branches were active against the Sarcoma 180.     

Comparison of subtraction arthrography, radionuclide arthrography and conventional plain radiography to assess loosening of total knee arthroplasty.

The value of plain radiographs, digital subtraction arthrography and radionuclide arthrography was analysed in 23 cases of failed total knee arthroplasty. The preoperative diagnosis was compared with the intraoperative assessment. Sensitivity, specificity and the positive and negative predictive value for assessing a loose component were determined separately for the femoral and tibial components. At revision we found 13 loose femoral and 12 loose tibial implants. In eight cases both components were unstable. Plain radiography had a sensitivity of 77% for loosening of the femoral and 83% for the tibial component; digital subtraction arthrography 77% for the femoral and 8% for the tibial component and radionuclide arthrography 31% and 8%. The specificity for plain radiography was 90% for the femoral and 72% for the tibial implant. For subtraction arthrography it was 50% and 82% and for subtraction arthrography 70% and 82%. Radiography had the highest positive and negative predictive values for both components compared with the other two techniques. As a diagnostic tool to detect implant loosening, plain radiography is the most effective in this study. Subtraction arthrography and radionuclide arthrography are not suitable for use as routine methods for detection of total knee arthroplasty loosening.     

Differential protein expression in anatomical zones of the prostate

The prostate has three anatomical zones: the peripheral (PZ), the transition (TZ), and the central (CZ) zone. It is proposed that the CZ may be of mesodermal origin, whereas the other two are of endodermal origin. Proteome patterns in the zones were characterized to test for differences. Cells were scraped from macroscopically normal areas of PZ, TZ, and CZ in radical prostatectomy specimens. After exclusion of samples with cancer or prostatic intraepithelial neoplasia, 18 cases remained for analysis. Cells were collected in a medium with protease inhibitors, and the protein material was prepared for two-dimensional gel electrophoresis. The proteins in spots that differed quantitatively between regions were identified via mass spectrometric fingerprinting of tryptic fragments and selected tandem mass spectrometry sequence analysis. Ten proteins with significant zonal differential expression were identified, eight with underexpression in the CZ versus the PZ and the TZ (arginase II, ATP synthase, cytokeratin 8, lamin A/C, peroxiredoxin 4, protein disulfide isomerase A3, tropomyosin, and vimentin), and two with overexpression in the CZ (peroxiredoxin 2 and creatine kinase B). The PZ and TZ, although differing in terms of incidence of cancer and hyperplasia, have epithelium with highly similar major protein expression profiles. However, the protein profile of the CZ differs from that of the other regions, suggesting functional differences.     

Involvement of the AP-1 adaptor complex in early steps of phagocytosis and macropinocytosis

The best described function of the adaptor complex-1 (AP-1) is to participate in the budding of clathrin-coated vesicles from the trans-Golgi network and endosomes. Here, we show that AP-1 is also localized to phagocytic cups in murine macrophages as well as in Dictyostelium amoebae. AP-1 is recruited to phagosomal membranes at this early stage of phagosome formation and rapidly dissociates from maturing phagosomes. To establish the role of AP-1 in phagocytosis, we made used of Dictyostelium mutant cells (apm1(-) cells) disrupted for AP-1 medium chain. In this mutant, phagocytosis drops by 60%, indicating that AP-1 is necessary for efficient phagocytosis. Furthermore, phagocytosis in apm1(-) cells is more affected for large rather than small particles, and cells exhibiting incomplete engulfment are then often observed. This suggests that AP-1 could participate in the extension of the phagocytic cup. Interestingly, macropinocytosis, a process dedicated to fluid-phase endocytosis and related to phagocytosis, is also impaired in apm1(-) cells. In summary, our data suggest a new role of AP-1 at an early stage of phagosome and macropinosome formation.     

Three-dimensional measurements of the pressure distribution in artificial joints with a capacitive sensor array

A spherically folded capacitive pressure sensor array is introduced and characterized. By placing the sensor array between the ball and the cavity of artificial joints, the pressure distribution within the joint was recorded with spatial resolution for different size matching between the ball and the cavity, for different directions of loading and for joints with incomplete cavities. The performance of the sensor array is analyzed, possible fields of application as well as its limitations are discussed.     

Restitution of defective glucose-stimulated insulin release of sulfonylurea type 1 receptor knockout mice by acetylcholine

Inhibition of ATP-sensitive K+ (K(ATP)) channels by an increase in the ATP/ADP ratio and the resultant membrane depolarization are considered essential in the process leading to insulin release (IR) from pancreatic beta-cells stimulated by glucose. It is therefore surprising that mice lacking the sulfonylurea type 1 receptor (SUR1-/-) in beta-cells remain euglycemic even though the knockout is expected to cause hypoglycemia. To complicate matters, isolated islets of SUR1-/- mice secrete little insulin in response to high glucose, which extrapolates to hyperglycemia in the intact animal. It remains thus unexplained how euglycemia is maintained. In recognition of the essential role of neural and endocrine regulation of IR, we evaluated the effects of acetylcholine (ACh) and glucagon-like peptide-1 (GLP-1) on IR and free intracellular Ca2+ concentration ([Ca2+]i) of freshly isolated or cultured islets of SUR1-/- mice and B6D2F1 controls (SUR1+/+). IBMX, a phosphodiesterase inhibitor, was also used to explore cAMP-dependent signaling in IR. Most striking, and in contrast to controls, SUR1-/-) islets are hypersensitive to ACh and IBMX, as demonstrated by a marked increase of IR even in the absence of glucose. The hypersensitivity to ACh was reproduced in control islets by depolarization with the SUR1 inhibitor glyburide. Pretreatment of perifused SUR1-/- islets with ACh or IBMX restored glucose stimulation of IR, an effect expectedly insensitive to diazoxide. The calcium channel blocker verapamil reduced but did not abolish ACh-stimulated IR, supporting a role for intracellular Ca2+ stores in stimulus-secretion coupling. The effect of ACh on IR was greatly potentiated by GLP-1 (10 nM). ACh caused a dose-dependent increase in [Ca2+]i at 0.1-1 microM or biphasic changes (an initial sharp increase in [Ca2+]i followed by a sustained phase of low [Ca2+]i) at 1-100 microM. The latter effects were observed in substrate-free medium or in the presence of 16.7 mM glucose. We conclude that SUR1 deletion depolarizes the beta-cells and markedly elevates basal [Ca2+]i. Elevated [Ca2+]i in turn sensitizes the beta-cells to the secretory effects of ACh and IBMX. Priming by the combination of high [Ca2+]i, ACh, and GLP-1 restores the defective glucose responsiveness, precluding the development of diabetes but not effectively enough to cause hyperinsulinemic hypoglycemia.     

Phosphodiesterase type 5 inhibition improves early memory consolidation of object information

The nitric oxide (NO)-cyclic GMP (cGMP) signaling pathway is assumed to play an important role in processes underlying learning and memory. We used phosphodiesterase type 5 (PDE5) inhibitors to study the role of cGMP in object- and spatial memory. Our results and those reported in other studies indicate that elevated hippocampal cGMP levels are required to improve the memory performance of rodents in object recognition and passive avoidance learning, but not in spatial learning. The timing of treatment modulates the effects on memory and strongly supports a role for cGMP in early stages of memory formation. Alternative explanations for the improved memory performance of PDE5 inhibitors are also discussed. Immunocytochemical studies showed that in vitro slice incubations with PDE5 inhibitors increase NO-stimulated cGMP levels mainly in hippocampal varicose fibers. Reviewing the available data on the localization of the different components of the NO-cGMP signaling pathway, indicates a complex interaction between NO and cGMP, which may be independent of each other. It is discussed that further studies are needed, immunocytochemical and behavioral, to better understand the cGMP-mediated molecular mechanisms underlying memory formation.