Pattern formation. Spiral cracks without twisting.
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Journal Article↗
Biomedical subjects
K Leung
Publications and source records attributed to K Leung.
Staining intensity of individual osteons correlated with elastic properties and degrees of mineralization.
Surface staining is widely used for histological studies involving undecalcified thick bone sections. Individual osteons, particularly newly formed ones stained with toluidine blue (TB), show various color intensities. We studied the correlations between TB color intensity and the differences in stiffness and degree of mineralization of individual osteons in undecalcified histological sections of goat tibial diaphysis, measured by scanning acoustic microscopy (SAM) and contact microradiography (CMR), respectively. Results showed that all three measurements correlated significantly with each other (r = 0.567 - 0.786; all P < or = 0.01). The TB surface staining intensity of individual osteons correlated better with the reflection coefficient (stiffness index) measured by SAM (r = 0.713) than with the aluminum step-wedge equivalent thickness measured on CMR micrographs (r = 0.567). The aluminum step-wedge equivalent thickness of individual osteons on CMR correlated slightly better with the SAM reflection coefficient (r = 0.786) than with the TB surface staining intensity (r = 0.713). The results of this study suggest that TB surface staining may be used as a simple method for indicating differences in stiffness and degree of mineralization in individual osteons in comparative histological studies.
Animals↗
Identification by 16S ribosomal RNA gene sequencing of an Enterobacteriaceae species with ambiguous biochemical profile from a renal transplant recipient.
Traditional ways of identification of bacteria by phenotypic characteristics cannot be used for non-cultivable organisms and organisms with unusual biochemical profiles. In this study, an Enterobacteriaceae was isolated in pure growth from the mid-stream urine of a 67-year old renal transplant recipient with urinary tract infection. Conventional biochemical tests did not reveal a pattern resembling any known member of the Enterobacteriaceae family. The Vitek system (GNI+) showed that it was 18% Leclercia adecarboxylata and 55% Klebsiella ozaenae; whereas the API system (20E) showed that it was 99.8% Rahnella aquatilis. 16S ribosomal RNA gene sequencing showed that there was 7 base differences between the isolate and Enterobacter cloacae, 18 base differences between the isolate and Enterobacter asburiae, 17 base differences between the isolate and Enterobacter cancerogenus, 35 base differences between the isolate and K. ozaenae, 27 base differences between the isolate and L. adecarboxylata, and 72 base differences between the isolate and R. aquatilis, indicating that the isolate most closely resembled a strain of E. cloacae. Identification of the organism in this study is important, as the choice of antibiotics would be radically different. In this case, cephalosporins should be avoided regardless of in-vitro susceptibility as cephalosporins are well-known to select for AmpC derepressed mutants in Enterobacter, and previous administration of third-generation cephalosporins is more likely to be associated with multidrug resistant Enterobacter isolates than is administration of antibiotics that do not include a third-generation cephalosporin.
Aged↗
Identification of Arcobacter cryaerophilus isolated from a traffic accident victim with bacteremia by 16S ribosomal RNA gene sequencing.
Traditional ways of identifying slow growing bacteria is slow and often difficult. In this study, a small, Gram-negative, facultative anaerobic, slow growing bacillus was isolated from the blood culture of a 7-year old traffic accident victim. The bacterium was non-hemolytic, catalase and oxidase positive. An attempt to use the Vitek system (GNI+) and the API system (20NE) to identify the strain was unsuccessful as the growth controls showed negative results. 16S ribosomal RNA gene sequencing showed that there was 1 base difference between the isolate and Arcobacter cryaerophilus (GenBank Accession no. U25805), 1 base difference between the isolate and A. cryaerophilus (GenBank Accession no. U34387), 10 base differences between the isolate and A. cryaerophilus (GenBank Accession no. L14624), 34 base differences between the isolate and A. butzleri (GenBank Accession no. U34386), 34 base differences between the isolate and A. butzleri (GenBank Accession no. U34387), and 38 base differences between the isolate and A. butzleri (GenBank Accession no. L14626), indicating that the isolate most closely resembled a strain of A. cryaerophilus. Identification of the isolate in our case by conventional methods was difficult, as the absence of a curved morphology has made it confused with other Gram-negative non-fermentative bacteria, and the slow growth rate has made it unidentifiable by both the Vitek and API systems. Although the exact source of infection and route of transmission in our case remains elusive, we speculate that the bacteria were transmitted through the respiratory tract while the boy was suffocated in the mud. The present report represents an example of showing the usefulness of 16S rRNA gene sequencing for identification of slow growing bacteria.
Accidents, Traffic↗
Studies on the metabolism of troglitazone to reactive intermediates in vitro and in vivo. Evidence for novel biotransformation pathways involving quinone methide formation and thiazolidinedione ring scission.
Therapy with the oral antidiabetic agent troglitazone (Rezulin) has been associated with cases of severe hepatotoxicity and drug-induced liver failure, which led to the recent withdrawal of the product from the U.S. market. While the mechanism of this toxicity remains unknown, it is possible that chemically reactive metabolites of the drug play a causative role. In an effort to address this possibility, this study was undertaken to determine whether troglitazone undergoes metabolism in human liver microsomal preparations to electrophilic intermediates. Following incubation of troglitazone with human liver microsomes and with cDNA-expressed cytochrome P450 isoforms in the presence of glutathione (GSH), a total of five GSH conjugates (M1-M5) were detected and identified tentatively by LC-MS/MS analysis. In two cases (M1 and M5), the structures of the adducts were confirmed by NMR spectroscopy and/or by comparison with an authentic standard prepared by synthesis. The formation of GSH conjugates M1-M5 revealed the operation of two distinct metabolic activation pathways for troglitazone, one of which involves oxidation of the substituted chromane ring system to a reactive o-quinone methide derivative, while the second involves a novel oxidative cleavage of the thiazolidinedione (TZD) ring, potentially generating highly electrophilic alpha-ketoisocyanate and sulfenic acid intermediates. When troglitazone was administered orally to a rat, samples of bile were found to contain GSH conjugates which reflected the operation of these same metabolic pathways in vivo. The finding that metabolism of the TZD ring of troglitazone was catalyzed selectively by P450 3A enzymes is significant in light of the recent report that troglitazone is an inducer of this isoform in human hepatocytes. The implications of these results are discussed in the context of the potential for troglitazone to covalently modify hepatic proteins and to cause oxidative stress through redox cycling processes, either of which may play a role in drug-induced liver injury.
Animals↗
Personality in cultural context: methodological issues.
Methodological issues in cultural and cross-cultural personality research are described. A taxonomy of these studies is presented, based on whether a study is exploratory or tests hypotheses, and whether or not contextual information is measured. Core methodological issues are bias and equivalence: a taxonomy and a brief overview of statistical procedures to examine equivalence are presented, with a focus on procedures for assessing structural equivalence (i.e., similarity of meaning of an instrument across cultures). Examples are given of studies in which cultural and cross-cultural approaches, often seen as antithetical, have been fruitfully integrated. Finally, multilevel models are described in which personality characteristics are examined at individual and cultural level.
Cross-Cultural Comparison↗
Inhibitory effects of nitric oxide on estrogen production and cAMP levels in rat granulosa cell cultures.
Previous studies demonstrated inhibitory effects of nitric oxide (NO) and cGMP on ovarian steroidogenesis. This study examined the effects of NO on estrogen levels and cAMP accumulation from immature cultured rat granulosa cells. Granulosa cells were incubated with media alone (control), FSH or FSH plus increasing concentrations of the NO generator, (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA/NO). While FSH increased estrogen levels 15-fold compared with controls, DETA/NO inhibited FSH-stimulated aromatase activity in a dose-dependent manner. Time-course studies revealed that the inhibitory effects of DETA/NO on aromatase activity persisted throughout the 72 h culture period. Treatment with DETA/NO also inhibited the stimulatory effects of forskolin on estrogen production, indicating that NO can influence steroidogenesis by actions downstream of the FSH receptor. Incubation of cells with FSH plus DETA/NO increased cGMP accumulation over 100-fold, compared with cells treated with media or FSH alone. In this regard, a cGMP analog mimicked the inhibitory effects of NO on FSH- and forskolin-stimulated estrogen production, indicating a potential mechanism of NO action. NO also decreased FSH-stimulated (cAMP) accumulation from cultured cells, indicating an antagonistic effect of NO on the second messenger mediating FSH actions. These findings demonstrate that NO inhibits estrogen production from rat granulosa cells, potentially reflecting actions on the second messengers cGMP and cAMP.
Animals↗
Pattern formation and selection in quasistatic fracture.
Fracture in quasistatically driven systems is studied by means of a discrete spring-block model. Developed from close comparison with desiccation experiments, it describes crack formation induced by friction on a substrate. The model produces cellular, hierarchical patterns of cracks, characterized by a mean fragment size linear in the layer thickness, in agreement with experiments. The selection of a stationary fragment size is explained by exploiting the correlations prior to cracking. A scaling behavior associated with the thickness and substrate coupling, derived and confirmed by simulations, suggests why patterns have similar morphology despite their disparity in scales.
Journal Article↗
Restricted expression of an adenoviral vector encoding Fas ligand (CD95L) enhances safety for cancer gene therapy.
Gene transfer of Fas ligand (CD95L) using adenoviral vectors has been shown to generate apoptotic responses and potent inflammatory reactions that can be used to induce the regression of malignancies in vivo, but these vectors also cause significant hepatotoxicity that may limit their clinical utility. Here we describe an adenoviral vector encoding CD95L with restricted gene expression that reduces its toxicity in vivo. Preclinical efficacy and gene expression studies of lineage-restricted CD95L adenoviral vectors were performed. To enhance its cytotoxicity and reduce potential systemic effects, a noncleavable CD95L was made by deleting a segment containing the cleavage site (CD95L deltaQP). Higher CD95L expression of this mutant was observed on the tumor cell surface, together with a reduction in the release of soluble CD95L. This CD95L cleavage mutant was then expressed under control of a smooth muscle-specific promoter, SM22apha, and analyzed for its ability to suppress the growth of tumors of smooth muscle origin in vivo. Growth of human leiomyosarcomas but not gliomas was inhibited after ADV gene transfer into tumor-bearing immunodeficient mice. In contrast to viral promoters, in which mortality was uniformly seen after injection of 10(12) particles, no significant hepatic injury or systemic toxicity was observed in mice, and the maximum tolerated dose was increased > or = 10- to 100-fold. These findings suggest that restricted specificity of adenoviral CD95L gene expression enhances the safety of this approach for cancer gene therapy.
Adenoviridae↗
Further refinement on the engineering of adenosine phosphorylase from purine nucleoside phosphorylase.
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Adenosine↗
Viability of competing field theories for the driven lattice gas
It has recently been suggested that the driven lattice gas should be described by an alternate field theory in the limit of infinite drive. We review the original and the alternate field theory, invoking several well-documented key features of the microscopics. Since the alternate field theory fails to reproduce these characteristics, we argue that it cannot serve as a viable description of the driven lattice gas. Recent results, for the critical exponents associated with this theory, are reanalyzed and shown to be incorrect.
Journal Article↗
Variation of measured sound speeds in gaseous and liquid air with temperature and pressure
Based on sound speeds in gaseous and liquid air measured by Younglove and Frederick [Int. J. Thermophys. 13(6), 1033-1041 (1992)], empirical equations for the computation of sound speeds in the above media at relatively smaller temperature and pressure ranges were derived. For gaseous air, over a temperature range from 200 to 300 K and pressure from 0.614 to 10.292 MPa, the maximum deviation between the measured sound speeds and those computed with the empirical equation is 56 ppm. For liquid air, over the ranges from 90 to 110 K for temperature and from 0.763 to 13.823 MPa for pressure, the corresponding deviation is 173 ppm.
Journal Article↗
Lack of functional receptors is the only barrier that prevents caprine arthritis-encephalitis virus from infecting human cells.
Barriers to replication of viruses in potential host cells may occur at several levels. Lack of suitable and functional receptors on the host cell surface, thereby precluding entry of the virus, is a frequent reason for noninfectivity, as long as no alternative way of entry (e.g., pinocytosis, antibody-dependent adsorption) can be exploited by the virus. Other barriers can intervene at later stages of the virus life cycle, with restrictions on transcription of the viral genome, incorrect translation and posttranslational processing of viral proteins, inefficient viral assembly, and release or efficient early induction of apoptosis in the infected cell. The data we present here demonstrate that replication of caprine arthritis-encephalitis virus (CAEV) is restricted in a variety of human cell lines and primary tissue cultures. This barrier was efficiently overcome by transfection of a novel infectious complete-proviral CAEV construct into the same cells. The successful infection of human cells with a vesicular stomatitis virus (VSV) G-pseudotyped Env-defective CAEV confirmed that viral entry is the major obstacle to CAEV infection of human cells. The fully efficient productive infection obtained with the VSV-G-protein-pseudotyped infectious CAEV strengthened the evidence that lack of viral entry is the only practical barrier to CAEV replication in human cells. The virus thus produced retained its original host cell specificity and acquired no propensity to propagate further in human cultures.
Animals↗
Asthmatic subjects symptomatically worse at work: prevalence and characterization among a general asthma clinic population.
STUDY OBJECTIVES: To assess the prevalence of a historical occupational component to asthma in an adult asthma clinic and to compare characteristics of asthmatic subjects with and without work-attributed symptoms. DESIGN: A retrospective review of data obtained from a physician-administered questionnaire, answers to which were obtained at the initial patient visit of asthmatic subjects, and which included specific questions regarding the relationship of work to symptoms. Chart review data were used to supplement information on workplace exposures and investigations. SETTING: A university-based secondary- and tertiary-referral asthma clinic. PATIENTS: Seven hundred thirty-one adult asthmatic subjects who were referred for assessment and management of asthma. INTERVENTIONS: Statistical analyses of asthmatic subjects with and without work-attributed symptoms and a determination, from chart review, of the likelihood of causes for symptomatic worsening of asthma at work. MEASUREMENTS AND RESULTS: Sixty percent of the patients (435) had adult onset of asthma, among whom 310 patients (71%) were employed at the time of their visit. Fifty-one patients reported their asthma to be worse at work (ie, 16% of adult-onset working asthmatic subjects). Sixteen of these patients (31%) had likely or possible sensitizer-induced occupational asthma (OA), and 49% likely had aggravation of underlying asthma. The other 20% of patients had possible OA or aggravation of underlying asthma at work. CONCLUSIONS: Adult-onset asthmatic subjects commonly report a worsening of asthma at work, more commonly on the basis of likely aggravation of underlying asthma than on the basis of likely or possible OA.
Adolescent↗
Derivation and biological characterization of a molecular clone of SHIV(KU-2) that causes AIDS, neurological disease, and renal disease in rhesus macaques.
Previously, we described the derivation of a pathogenic strain of simian-human immunodeficiency virus (SHIV(KU-2)) consisting of the tat, rev, vpu, and env genes of HIV-1 (strain HXB2) in a genetic background of SIV(mac)239 that causes AIDS and productive infection of the CNS in rhesus macaques (Macca mulatta) (Raghavan et al., 1997, Brain Pathol. 7, 851-861). We report here on the characterization of a molecular clone of SHIV(KU-2), designated SHIV(KU-2MC4), that caused CD4(+) T cell loss as well as neurological and renal disease in macaques. DNA sequence analysis of selected SIV regions of SHIV(KU-2MC4) revealed 10 nucleotide changes in the LTR, whereas Gag, Vif, Vpr, Vpx, and Nef had 1, 1, 1, 2, and 13 predicted amino acid substitutions, respectively, compared to SIV(mac)239. DNA sequence analysis of HIV-1 derived regions of SHIV(KU-2MC4) revealed 2, 1, 2, and 18 predicted amino acid substitutions in the Tat, Rev, Vpu, and Env proteins, respectively, when compared to SHIV-4. Unlike the parental SHIV-4, which is not tropic for macrophages, SHIV(KU-2MC4) replicated efficiently in macrophage cultures as determined by p27 assays. However, despite the numerous changes in the Env protein and newly acquired tropism for macrophages, SHIV(KU-2MC4), like the parental SHIV-4, used CXCR4 exclusively as its coreceptor for entry into susceptible cells. Inoculation of SHIV(KU-2MC4) into two rhesus macaques resulted in severe infection in which the numbers of circulating CD4(+) T cells in the blood declined rapidly by 2 weeks postinoculation and virus producing cells in the peripheral blood mononuclear cells were identified throughout the course of infection. At the time of euthanasia (20 and 22 weeks), both macaques had lost a significant amount of weight and had no circulating CD4(+) T cells. In addition, one macaque developed intension tremors and uncoordinated movements. Virological examination of tissues at necropsy revealed active virus replication in both lymphoid and nonlymphoid tissues such as the lung and brain. Histological examination revealed that the induced immunodeficiency was associated with lymphoid depletion of the lymph nodes and spleen, opportunistic infections, lentiviral encephalitis, and severe glomerulosclerosis of the kidney. This molecular clone will serve as the basis for analyzing the molecular determinants through which SHIV(KU-2) causes severe CD4(+) T cell loss, neurological disease, and SHIV nephropathy in rhesus macaques.
AIDS Dementia Complex↗
Thiazole-derived potent, highly bioavailable short duration growth hormone secretagogues.
Replacement of the phenyl in 3 with a 2-pyridyl or 4-thiazolyl group resulted in increased potency in the rat pituitary cell GH release assay and in beagles.
Animals↗
Identification of a fimbriae-associated haemagglutinin from Prevotella intermedia.
Prevotella intermedia, a putative periodontopathic microorganism, possesses various types of fimbriae (surface appendages). Some of these surface structures mediate the adherence of the organism to several mammalian erythrocytes, resulting in the agglutination of the erythrocytes. Prevotella intermedia fimbriae were solubilized and separated by preparative SDS gel electrophoresis followed by preparative isoelectric focusing to determine which fimbrial component(s) were responsible for the haemagglutinating activity exhibited by these bacteria. Heat treatment of isolated fractions which exhibited haemagglutinating activity revealed the presence of two types of haemagglutinating activity which were either heat sensitive or resistant. Analysis of isolated fractions, which exhibited haemagglutinating activity that were heat labile, by Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a 3.8 kD peptide. The purified peptide had a pI of 4.8-5.0. Examination of fractions containing this peptide by electron microscopy showed the presence of fimbriae. Immunogold labelling showed that mouse antibodies raised against the 3.8 kD peptide bound specifically and extensively to P. intermedia fimbriae. It appears that this peptide is a fimbriae-associated haemagglutinin and may represent a major constituent of the fimbriae. Further, fractions exhibiting haemagglutinating activity that were heat resistant, which were recovered at a pH of 3.5 in preparative isoelectric focusing of fimbrial samples, did not possess any detectable major protein bands as shown by analytical gel electrophoresis. However, silver stained gels for the detection of lipopolysaccharide (LPS) revealed the presence of LPS-like components in these fractions. In addition, LPS isolated from whole cells showed a similar electrophoretic pattern and exhibited the haemagglutinating activity that was heat resistant. The results of this study strongly suggest that P. intermedia possesses the machinery to agglutinate erythrocytes, which may be a contributing factor in its colonization in vivo.
Animals↗
Applications of the green fluorescent protein as a molecular marker in environmental microorganisms.
In this review, we examine numerous applications of the green fluorescent protein (GFP) marker gene in environmental microbiology research. The GFP and its variants are reviewed and applications in plant-microbe interactions, biofilms, biodegradation, bacterial-protozoan interactions, gene transfer, and biosensors are discussed. Methods for detecting GFP-marked cells are also examined. The GFP is a useful marker in environmental microorganisms, allowing new research that will increase our understanding of microorganisms in the environment.
Biotechnology↗