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Immediate detection of postreconstructive ischaemia by intra-operative Doppler ankle pressure index monitoring during aortic reconstructions.

From March 1980 to February 1988, 368 aorto-iliac or aorto-femoral bilateral reconstructions were performed for aneurysmal or occlusive disease. In order to demonstrate early postreconstructive ischaemic complications, the intra-operative Doppler Ankle Pressure Index (API) was measured immediately before reconstruction (PRE), just after declamping (DEC) and 5, 15, 30 and 45 min thereafter. Each limb of the reconstruction (736) was considered individually and subdivided as follows: Group (A) 705 immediately successful (96% of limbs and 92% of patients); Group (B) 22 (3% of limbs and 5% of patients) in which, on the grounds of API data, an ischaemic complication was regarded as imminent and treated by graft revision or a peripheral thromboembolectomy (intra-operative corrections); Group (C) 9 (1% of limbs and 2% of patients) in which the diagnosis of ischaemic complications was made postoperatively when the patients underwent successful reoperation. All 736 reconstructions were patent and functional at discharge of the patient. By a retrospective analysis the intra-operative APIs were studied and the features (single or associated) suggesting an ischaemic complication examined. These were: (1) no flow at the time of declamping or its disappearance during the operation (10 cases, 1.25% of limbs) all detected and successfully corrected intra-operatively.(ABSTRACT TRUNCATED AT 250 WORDS)

Ankle↗

A comparison of methods used in species identification of coagulase-negative staphylococci isolated from the milk of sheep.

Thirty reference strains of staphylococci, 76 strains isolated from confirmed cases of subclinical mastitis and 10 strains from the teat tip skin and the mouth of sucking lambs were assigned to species using four biochemical methods. These were the commercially available micromethods API Staph and Staph-Zym; the specialized laboratory method Rosco Set and Reactions in Standard Laboratory Culture Media (SLCM). The Rosco Set assigned species to all the strains, the SLCM to 96.5%, the Staph-zym to 86.1% and the API Staph to 81.4%. API Staph and the SLCM favoured the 'aureus' group with the 'epidermidis' second, while the reverse was true for the other two methods. All the identification methods assigned the same species to only 29% of the tested isolates, causing considerable variation in the number of species identified within each group. This variation indicates that reported species prevalence from cases of ovine mastitis should be viewed with caution.

Animals↗

Characterization of crude oils using fluorescence lifetime data.

The average fluorescence lifetimes of nine North Sea crude oils with API gravities of between 20 and 51 were measured using a modular, filter based, instrument developed in-house. Two pulsed light emitting diode (LED) excitation sources (460 and 510 nm) were used to excite fluorescence, the lifetime of which was measured at a range of emission wavelengths. Fluorescence lifetimes were found to vary from 1.8 to 8.2 ns with confidence intervals of +/- 0.11 ns. The average lifetimes at all emission wavelengths were linearly correlated with API gravity and with aromatic concentration with the best results being obtained with the 460 nm excitation source. Predictive models with an accuracy of +/- 7.6 API degrees were generated using partial least-squares methods from average fluorescence lifetimes measured at an emission wavelength of 500 nm using 460 nm excitation. A better correlation was found between the aromatic concentration of the oils and the ratio of the average fluorescence lifetimes at measured at 550 and 650 nm using 460 nm excitation. This led to a quantitative model with an accuracy of +/- 5.4% for aromatic concentration.

Biophysical Phenomena↗

Biochemical and serological investigations on clinical isolates of klebsiella.

A series of 925 clinical isolates of klebsiella was examined by serological and biochemical typing. To perform serological typing (capsular swelling) 77 capsular antisera were prepared, tested against the type strains and grouped in 13 pools. With this serotyping method 80% of the cultures were typable and 63 distinct types could be recognized. All strains were typable biochemically by means of the numerical coding system of the API-20E system supplemented by digits derived from 15 additional conventional biochemical tests. With the API-20E system 24 different biotypes could be distinguished whereas the combination of API-20E and the 15 additional tests produced 93 biotypes. Maximum discrimination of strains was achieved by the combination of serological and biochemical typing (256 bioserotypes). The reproducibility, typability and discriminating power of the biotyping system was not inferior to serotyping. For epidemiological purposes biotyping can replace serotyping of Klebsiella species, especially in laboratories less well equipped.

Antibodies, Bacterial↗

Evolution of new function in the GTP cyclohydrolase II proteins of Streptomyces coelicolor.

The genome sequence of Streptomyces coelicolor contains three open reading frames (sco1441, sco2687, and sco6655) that encode proteins with significant (>40%) amino acid identity to GTP cyclohydrolase II (GCH II), which catalyzes the committed step in the biosynthesis of riboflavin. The physiological significance of the redundancy of these proteins in S. coelicolor is not known. However, the gene contexts of the three proteins are different, suggesting that they may serve alternate biological niches. Each of the three proteins was overexpressed in Escherichia coli and characterized to determine if their functions are biologically overlapping. As purified, each protein contains 1 molar equiv of zinc/mol of protein and utilizes guanosine 5'-triphosphate (GTP) as substrate. Two of these proteins (SCO 1441 and SCO 2687) produce the canonical product of GCH II, 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate (APy). Remarkably, however, one of the three proteins (SCO 6655) converts GTP to 2-amino-5-formylamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate (FAPy), as shown by UV-visible spectrophotometry, mass spectrometry, and NMR. This activity has been reported for a GTP cyclohydrolase III protein from Methanocaldococcus jannaschii [Graham, D. E., Xu, H., and White, R. H. (2002) Biochemistry 41, 15074-15084], which has no amino acid sequence homology to SCO 6655. Comparison of the sequences of these proteins and mapping onto the structure of the E. coli GCH II protein [Ren, J., Kotaka, M., Lockyer, M., Lamb, H. K., Hawkins, A. R., and Stammers, D. K. (2005) J. Biol. Chem. 280, 36912-36919] allowed identification of a switch residue, Met120, which appears to be responsible for the altered fate of GTP observed with SCO 6655; a Tyr is found in the analogous position of all proteins that have been shown to catalyze the conversion of GTP to APy. The Met120Tyr variant of SCO 6655 acquires the ability to catalyze the conversion of GTP to APy, suggesting a role for Tyr120 in the late phase of the reaction. Our data are consistent with duplication of GCH II in S. coelicolor promoting evolution of a new function. The physiological role(s) of the gene clusters that house GCH II homologues will be discussed.

Bacterial Proteins↗

Synthesis and characterization of the face-sharing bioctahedral [Mo2O6F3]3- anion.

The face-sharing bioctahedral molybdenum(VI) oxide fluoride anion [Mo2O6F3]3- has been isolated in the new compound [Cu(3-apy)4]3(Mo2O6F3)2 (3-apy = 3-aminopyridine) and has been characterized by experimental and computational techniques. Single-crystal X-ray diffraction studies show that the structure of the [Mo2O6F3]3- anion resembles two distorted face-sharing octahedra, each with three short terminal metal-ligand bonds and three long metal-ligand-metal bridging interactions. Aspects of the electronic structure, as well as geometric comparisons of the bond lengths and angles in [Mo2O6F3]3- with those in the similarly distorted [MoO3F3]3- anion, suggest that the six terminal ligand positions of the confacial bioctahedra are occupied exclusively by oxide ligands and that the three bridging sites are occupied by fluorides. Crystal data for [Cu(3-apy)4]3(Mo2O6F3)2: trigonal space group R3 (No. 148) with hexagonal axes of a = 13.881(1) A and c = 31.783(3) A (Z = 3).

Journal Article↗

Atmospheric pressure ionization LC-MS-MS determination of urushiol congeners.

This paper describes atmospheric pressure ionization (API) LC-MS-MS determination of urushiols, 3-n-alkenyl- and -alkyl-substituted catechols responsible for poison oak dermatitis. Urushiol was isolated from Western poison oak according to the method of Elsohly et al. (1) (J. Nat. Prod. 1982, 45, 532-538)-the purified preparation contained C(17)- and C(15)-substituted urushiols with zero, one, two, and three double bonds as determined from GC-MS analysis of trimethylsilyl derivatives. Urushiol mixtures were separated on a C(18) reversed phase HPLC column with a methanol-water gradient with urushiols eluting in 100% methanol. Atmospheric pressure chemical ionization (APCI) produced primarily [M - H](-) and MH(+) molecule ions. Electrospray ionization (ESI) yielded [M - H](-) and adduct ions including [M + Cl](-). Daughter ions of [M - H](-) included quinoid radical anions ([M - H - H(2)](-) and m/z 122(-)) and a benzofuran phenate (m/z 135(-)). A suite of hydrocarbon fragments were produced by collision-induced dissociation of MH(+) directly or via an intermediate [MH - H(2)O](+) daughter ion. Six urushiol congeners, one not previously reported in poison oak, were determined by negative ion API-LC-MS-MS with detection limits of approximately 8 pg/microL (ESI) and approximately 800 pg/microL (APCI). API-LC-MS-MS was used to determine urushiol in surface wipes, air samples, and plant materials.

Atmospheric Pressure↗

Nereistoxin and cartap neurotoxicity attributable to direct block of the insect nicotinic receptor/channel.

Nereistoxin (NTX) (4-dimethylamino-1,2-dithiolane) is the naturally occurring prototype for cartap [the bis(thiocarbamate) derivative of the NTX dithiol], which is generally regarded as a proinsecticide reverting to NTX. The aim of this study is to define the target site(s) for dithiolanes and dithiol esters. The affinity of [(3)H]NTX was not suitable for binding assays with honeybee (Apis mellifera) head membranes. However, NTX and cartap are equally potent, direct-acting, and competitive displacers of [(3)H]thienylcyclohexylpiperidine binding at the noncompetitive blocker (NCB) site of the Apis nicotinic acetylcholine receptor (nAChR)/channel. NTX also binds at the Apis [(3)H]imidacloprid agonist site, but cartap does not. As candidate metabolic pathways, sequential N-desmethylation and S-oxidation of NTX progressively reduce its potency at the NCB site and toxicity to houseflies. A P450 inhibitor reduces the toxicity of NTX and enhances it with cartap. Surprisingly, cartap is not just a pro-NTX but instead directly induces inhibitory neurotoxicity by blocking the nAChR/channel, whereas NTX may have dual NCB and agonist targets.

Animals↗

Extracellular enzymatic activity of Fusarium section Liseola isolates.

Extracellular enzymatic activity of Fusarium species from section Liseola was determined by the Api-Zym test, a semiquantitative method for assaying different cellular enzymes. This enzyme testing system was evaluated for possible use as a species identification aid. Twenty-two strains of Fusarium section Liseola were used: 8 of F. moniliforme, 6 of F. proliferatum, 6 of F. subglutinans and 2 of F. anthophilum. The optimal spore concentration was also determined for maximal activity in Api-Zym assay. According to our findings, the species showed little difference in Api-Zym enzymatic profiles, being not suitable for identification of these species. Only F. subglutinans and F. anthophilum showed a different enzymatic pattern.

Journal Article↗

Racial/ethnic differences in caregiver strain and perceived social support among parents of youth with emotional and behavioral problems.

This study examined several hypotheses related to racial/ethnic variations in caregiver strain and perceived social support among parents caring for children with emotional/behavioral problems. A subsample of youth from the Patterns of Care (POC) study, which drew a stratified random sample of high-risk youth active to 1 of 5 public sectors of care, was used to test these hypotheses. When controlling for youth age, parental education, presence in an alcohol/drug or mental health (ADM) sector, and severity of child problems, African Americans reported significantly lower caregiver strain then did non-Hispanic Whites (NHWs), whereas Asian/Pacific Islanders (APIs) and Latinos did not differ significantly from NHWs. Contrary to prediction, both African Americans and APIs reported significantly lower perceived social support than NHWs, whereas Latinos did not differ significantly from NHWs. When perceived social support was controlled, the tendency of African American and API parents to report lower caregiver strain than NHWs became more pronounced. Thus, differences in perceived social support did not explain African Americans' lower caregiver strain. Other cultural variables such as familism and tolerance should be examined to identify processes that result in reduced perception of caregiver strain among African American parents caring for children with emotional/behavioral problems.

Adolescent↗

Inhibition of AKT survival pathway by a small molecule inhibitor in human endometrial cancer cells.

The PTEN (phosphatase and tensin homolog deleted on chromosome 10) tumour suppressor is mutated in 40-50% of human endometrial cancers. PTEN exerts its effects in part via inhibition of the antiapoptotic protein AKT. We demonstrate that two endometrial cancer cell lines that harbour PTEN mutations, Ishikawa and RL95-2, have high levels of phosphorylated AKT and high AKT kinase activity. Two additional endometrial cancer cell lines that express wild-type PTEN, Hec1A and KLE, have little phosphorylated AKT and minimal demonstrable AKT kinase activity. We tested a potential inhibitor of the AKT pathway, API-59CJ-OMe, in these four cell lines. We found that API-59CJ-OMe inhibits AKT kinase activity and induces apoptosis in the Ishikawa and RL95-2 cell lines with high AKT activity, but has little effect on Hec1A and KLE cells without AKT activity. API-59CJ-OMe may therefore have therapeutic potential for those endometrial cancers that harbour PTEN mutations and AKT activation.

Apoptosis↗

Effects of a mechanical interdental cleaning device on oral hygiene in patients with lingual brackets.

This study was aimed at determining the influence of a battery-operated interdental cleaning device (icd) (WaterPik Flosser) on the oral hygiene of 32 female right-handed patients (mean age 25.9 years) with lingual brackets in the upper (n = 29) and or in the lower arch (n = 25). Approximal plaque index (API) and bleeding on probing (BOP) were recorded at the lingual surfaces by a single blinded examiner before application (t0), on average 38.6 days after (t1), and again on average 46.0 days after (t2) the application of the icd. The patients used the icd once a day in the second and fourth quadrants only (icdq). In all quadrants (icdq and non-icd quadrants [n-icdq]), oral hygiene was performed with a manual toothbrush. Of the patients enrolled in the study, 96.9% found the icd subjectively very helpful to moderately helpful for cleaning their teeth and 65.6% had the subjective impression that their teeth were cleaner with the appliance. Despite those positive subjective assessments, an objective comparison of the icdqs with the n-icdqs revealed no statistically significant differences in the mean changes in API and BOP from t0 to t1, from t0 to t2, or from t1 to t2. Because there were spectacular improvements in API in all quadrants, the improvements could be interpreted as an outcome of the instruction and motivation given to the patients, the increasing awareness of oral hygiene, and the greater skill in using the toothbrush in the course of time.

Adolescent↗

Distribution of enterococci in Hong Kong.

OBJECTIVES: To study the distribution of hospital isolates of enterococci from urines, bile, blood and body fluids and to evaluate different methods for the identification of enterococci. METHODS: Enterococci isolated from urine, bile, blood and body fluids collected during 1997 and 1998 were identified by polymerase chain reaction (PCR), API 20 Strep and conventional biochemical tests. RESULTS: A total of 498 non-duplicate enterococci were studied: 398 and 43 isolates from urine and bile, respectively, 49 from blood, two from cerebrospinal fluid and six from body fluids. Both API 20 Strep and PCR gave the same identification results for 240 Enterococcus faecalis isolates, 45 E. faecium isolates and one isolate each of E. gallinarum and E. Casseliflavus. These isolates were re-defined by conventional biochemical tests. PCR could correctly identify 303 (98%) isolates while API 20 Strep could only correctly identify 287 (93%) isolates (99% of E. faecalis and 57-87% of the other Enterococcus sp.). Thus, PCR was used in the identification of the remaining isolates and the identity of isolates other than E. faecalis was subsequently confirmed by biochemical tests. CONCLUSIONS: The majority of enterococci isolated was E. faecalis (81%) while only 15% were E. faecium and 4% the other enterococcal species. PCR could correctly identify E. faecalis while the identity of other enterococcal species had to be confirmed by biochemical tests.

Bacterial Typing Techniques↗

Evaluation of the location of the anus by a modified technique in the neonate.

PURPOSE: The aim of the current study was to bring to notice the anterior displacement of the anus and to recommend the measurement of anal position index in the neonate by a modified method. METHODS: Sixty newborns (34 girls and 26 boys) were taken into study, and the anal position index (API), which is the ratio of anus-fourchette (scrotum) distance to coccyx fourchette (scrotum) distance, was measured. To obtain the measurement, a transparent adhesive tape was placed along the midline on the long axis, covering the anus. The upper and lower tips and the center of the anal circle was marked and measured using a caliber. RESULTS: API was found as 0.46 (SD +/- 0.08) and 0.53 (SD +/- 0.05) in female and male neonates, respectively. Because an index of 0.34 in girls and 0.46 in boys are considered abnormal, the 3 female babies in the study group with API indices of 0.18, 0.28, and 0.33 were subjected to further examination. The abnormality was seen not to be rare in the Aegean region. CONCLUSIONS: An abnormal index alone cannot be the sole cause of constipation mentioned in the literature and therefore not an indication for operation. Anal position index in the neonates could be measured more accurately by the current modified method. If an anterior location of the anus is found early in infancy the baby should undergo follow-up accordingly.

Anal Canal↗

Existence of a rhamnogalacturonan II-like region in bioactive pectins from medicinal herbs.

Unusual component sugars such as 2-methylfucose (2-Me-Fuc), 2-methylxylose (2-Me-Xyl), apiose (Api), and aceric acid (AceA) are contained in the bioactive pectins from Bupleurum falcatum, Glycyrrhiza uralensis, and Angelica acutiloba, but not in the other bioactive pectic heteroglycans and arabinogalactans from Chinese and Japanese herbs tested. Each pectin was digested with endo-alpha-(1-->4)-polygalacturonase, and gave two enzyme-resistant fractions, PG-1 (rhamnogalacturonan core with neutral sugar side-chains) and PG-2, and an oligogalacturonide fraction (PG-3) by gel filtration on Bio-gel P-30. The PG-2 fractions commonly consisted of unusual sugars such as 2-Me-Fuc, 2-Me-Xyl, Api, AceA, 3-deoxy-D-lyxo-heptulosaric acid (Dha), and 3-deoxy-D-manno-2-octulosonic acid (Kdo) in addition to Rha, Fuc, Ara, Xyl, Man, Gal, Glc, GalA, and GlcA. Lithium degradation of each PG-2 gave a pentosyl-->6-deoxyhexosyl-->6-deoxyhexosyl-->pentitol fragment as a major oligosaccharide in addition to some neutral di- to trisaccharide alditols. Methylation analysis of the lithium degradation products from each PG-2 also indicated that these oligosaccharide alditols mainly consisted of terminal Rha, Araf, Fuc, Xyl, and Gal, 4-linked Rha, 3-linked Fuc, and 3'-linked Api. HPLC analysis showed that PG-2 had molecular heterogeneity. These results indicate that the bioactive pectins from medicinal herbs commonly consist of the minor KDO-containing region which resembles the rhamnogalacturonan II in plant cell walls.

Carbohydrate Sequence↗

Reconstitution of invertebrate glutamate receptor function depends on stargazin-like proteins.

alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors (AMPARs) are a major subtype of ionotropic glutamate receptors (iGluRs) that mediate rapid excitatory synaptic transmission in the vertebrate brain. Putative AMPARs are also expressed in the nervous system of invertebrates. In Caenorhabditis elegans, the GLR-1 receptor subunit is expressed in neural circuits that mediate avoidance behaviors and is required for glutamate-gated current in the AVA and AVD interneurons. Glutamate-gated currents can be recorded from heterologous cells that express vertebrate AMPARs; however, when C. elegans GLR-1 is expressed in heterologous cells, little or no glutamate-gated current is detected. This finding suggests that other receptor subunits or auxiliary proteins are required for function. Here, we identify Ce STG-1, a C. elegans stargazin-like protein, and show that expression of Ce STG-1 together with GLR-1 and the CUB-domain protein SOL-1 reconstitutes glutamate-gated currents in Xenopus oocytes. Ce STG-1 and homologues cloned from Drosophila (Dro STG1) and Apis mellifera (Apis STG1) have evolutionarily conserved functions and can partially substitute for one another to reconstitute glutamate-gated currents from rat, Drosophila, and C. elegans. Furthermore, we show that Ce STG-1 and Apis STG1 are primarily required for function independent of possible roles in promoting the surface expression of invertebrate AMPARs.

Amino Acid Sequence↗

A peptide template as an allosteric supramolecular catalyst for the cleavage of phosphate esters.

The heptapeptide H-Iva-Api-Iva-ATANP-Iva-Api-Iva-NHCH(3) (P1a), where Iva is (S)-isovaline, Api is 4-amino-4-carboxypiperidine, and ATANP is (S)-2-amino-3-[1-(1,4,7-triazacyclononane)]propanoic acid, has been synthesized. Its conformation in aqueous solution is essentially that of a 3(10)-helix. By connecting three copies of P1a to a functionalized Tris(2-aminoethyl)amine (Tren) platform a new peptide template, [T(P1)(3)], was obtained. This molecule is able to bind up to four metal ions (Cu(II) or Zn(II)): one in the Tren subsite and three in the azacyclononane subunits. The binding of the metals to the Tren platform induces a change from an open to a closed conformation in which the three short, helical peptides are aligned in a parallel manner with the azacyclonane units pointing inward within the pseudocavity they define. T(P1)(3) shows a peculiar behavior in the transphosphorylation of phosphate esters; the tetrazinc complex is a catalyst of the cleavage of 2-hydroxypropyl-p-nitrophenyl phosphate (HPNP), whereas the free ligand is a catalyst of the cleavage of an oligomeric RNA sequence with selectivity for pyrimidine bases. In the case of HPNP, Zn(II) acts as a positive allosteric effector by enhancing the catalytic efficiency of the system. In the case of the polyanionic RNA substrate, Zn(II) switches off the activity, thus behaving as a negative allosteric regulator. It is suggested that the opposite behavior of the catalyst induced by Zn(II) is associated with the change of conformation of the Tren platform, and consequently of the relative spatial disposition of the three linked peptides, that occurs after binding of the metal ion.

Allosteric Site↗

Cytoplasm-to-vacuole targeting and autophagy employ the same machinery to deliver proteins to the yeast vacuole.

The vacuolar protein aminopeptidase I (API) uses a novel cytoplasm-to-vacuole targeting (Cvt) pathway. Complementation analysis of yeast mutants defective for cytoplasm-to-vacuole protein targeting (cvt) and autophagy (apg) revealed seven overlapping complementation groups between these two sets of mutants. In addition, all 14 apg complementation groups are defective in the delivery of API to the vacuole. Similarly, the majority of nonoverlapping cvt complementation groups appear to be at least partially defective in autophagy. Kinetic analyses of protein delivery rates indicate that autophagic protein uptake is induced by nitrogen starvation, whereas Cvt is a constitutive biosynthetic pathway. However, the machinery governing Cvt is affected by nitrogen starvation as targeting defects resulting from API overexpression can be rescued by induction of autophagy.

Aminopeptidases↗