Isolation of leptin-binding peptides from a random peptide phage library

Leptin plays a role in regulating the body weight in mice. Injection of recombinant mouse leptin expressed in Escherichia coli reduced the food intake and body weight in normal, ob/ob and diet-induced obesity mice. Hyperglycemia, hyperinsulinemia and hypothermia can also be corrected in ob/ob mice after leptin injection. Leptin is a 16-kDa secretory protein comprising 167 amino acids produced in adipose tissue and is secreted to blood stream. In this study, a recombinant mouse leptin was generated and purified from a baculovirus expression system. This protein was used to identify putative ligands using a phage library of random peptides. Three leptin-binding phage clones were found, which were characterized by DNA sequencing and ELISA methods. The amino acid sequences of the reactive peptides are: LAYCSDPVRCLVWWY, MFWlSAVSFVDHALV and LVLVLSAFLCCGVG. All three clones bound to recombinant human and mouse leptins. These peptides may be useful tools to study leptin-receptor interaction, food intake and body weight regulation.postprin

Combined Rapid (TUBEX) Test for Typhoid-Paratyphoid A Fever Based on Strong Anti-O12 Response: Design and Critical Assessment of Sensitivity

Rapid diagnostics can be accurate but, often, those based on antibody detection for infectious diseases are unwittingly underrated for various reasons. Herein, we described the development of a combined rapid test for two clinically-indistinguishable bacterial diseases, typhoid and paratyphoid A fever, the latter fast emerging as a global threat. By using monoclonal antibodies (mAbs) to bacterial antigens of known chemical structures as probes, we were able to dissect the antibody response in patients at the level of monosaccharides. Thus, a mAb specific for a common lipopolysaccharide antigen (O12) found in both the causative organisms was employed to semi-quantify the amounts of anti-O12 antibodies present in both types of patients in an epitope-inhibition particle-based (TUBEX) immunoassay. This colorimetric assay detected not only anti-O12 antibodies that were abundantly produced, but also, by steric hindrance, antibodies to an adjoining epitope (O9 or O2 in the typhoid or paratyphoid bacillus, respectively). Sensitivity and, particularly, reaction intensities, were significantly better than those obtained using an anti-O9 or anti-O2 mAb-probe in the examination of paired sera from 22 culture-confirmed typhoid patients (sensitivity, 81.8% vs 75.0%) or single sera from 36 culture-confirmed paratyphoid patients (52.8% vs 28.6), respectively. Importantly, sensitivity was better (97.1% for typhoid, 75.0% for paratyphoid) if allowance was made for the absence of relevant antibodies in certain specimens as determined by an independent, objective assay (ELISA) — such specimens might have been storage-denatured (especially the older paratyphoid samples) or procured from non-responders. Benchmarking against ELISA, which revealed high concordance between the two tests, was useful and more appropriate than comparing with culture methods as traditionally done, since antibody tests and culture target slightly different stages of these diseases. Paired sera analysis was insightful, revealing 64% of typhoid patients who had no change in antibody titer over 4–16 days, and 14% with no IgM-IgG class-switching

Optimising antimicrobial prescription in hospitals by introducing an antimicrobial stewardship programme in Hong Kong: Consensus statement

Objective. To discuss the implementation of an 'antimicrobial stewardship programme' as a means to improve the quality of antimicrobial use in a hospital setting in Hong Kong. Participants. Consensus working group on 'antimicrobial stewardship programme', The Scientific Committee on Infection Control, Centre for Health Protection, Department of Health, comprised 11 experts. The remit of the working group was to discuss the rationale and requirement for optimising antimicrobial prescriptions in hospitals by the introduction of an 'antimicrobial stewardship programme'. Evidence. PubMed articles, national and international guidelines, and abstracts of international meetings published between January 2000 and December 2004 on programmes for improving the use of antimicrobials in hospitals. Only English medical literature was reviewed. Consensus process. Data search was performed independently by three members of the working group. They met on three occasions before the meeting to discuss all collected articles. A final draft was circulated to the working group before a meeting on 3 January 2005. Five commonly asked questions about an 'antimicrobial stewardship programme' were selected for discussion by the participants. Published information on the rationale, components, outcome measures, advantages, and disadvantages of the programme was reviewed. Recent unpublished data from local studies of an 'antimicrobial stewardship programme' were also discussed. The timing, potential problems, and practical issues involved in the implementation of an 'antimicrobial stewardship programme' in Hong Kong were then considered. The consensus statement was circulated to and approved by all participants. Conclusion. The continuous indiscriminate and excessive use of antimicrobial agents promotes the emergence of antibiotic-resistant organisms. Antimicrobial resistance substantially raises already-rising health care costs and increases patient morbidity and mortality. Pattern of prescriptions in hospitals can be improved through the implementation of an 'antimicrobial stewardship programme'. A 'universal' and 'continuous' 'antimicrobial stewardship programme' should now be established in Hong Kong hospitals.published_or_final_versio

Assessment of nine markers for phylogeny, species and haplotype identification of Kappaphycus species and Eucheuma denticulatum (Solieriaceae, Rhodophyta)

Molecular studies have contributed to the taxonomy of carrageenan-producing Kappaphycus spp. and Eucheuma denticulatum. However, unresolved species complexes and the lack of standardization in the use of genetic markers impede the identification of specimens and the delineation of a robust taxonomic framework. Here, nine molecular markers (cox1, cox2–3 spacer, cox2, cox3, COB, ITS, psbA, UPA and rbcL) were used to generate a multilocus phylogeny for 113 fresh eucheumatoid samples and four herbarium specimens. Analyses of species delineation and genetic distances confirmed the monophyly of currently accepted taxa. These analyses suggest that clades previously reported as K. striatus KS1 and KS2 are conspecific, and that E. denticulatum EDA ‘spinosum’ and EDB ‘endong/cacing’ are also conspecific. The results also unveiled possible new taxa from Hawaii and Indonesia. Each molecular marker and combinations thereof were assessed with regard to species identification, ease of amplification and sequencing, and haplotype characterization. All genetic markers recorded at least 94% success in the amplification and sequencing of fresh specimens, with cox1 being the most phylogenetically informative. Automatic partitioning, phylogenetic and tree-based assessments showed cox1, cox2–3 spacer, cox2 and rbcL were able to correctly identify species while cox1+ rbcL, COB+rbcL, cox2+ rbcL or cox1+ COB+rbcL trees best represented the phylogeny with consistently high nodal support. Among individual markers, cox1 identified the greatest number of haplotypes, while UPA, partial rbcL (750 bp), ITS, cox3 and cox2–3 spacer were able to retrieve information from herbarium specimens of 12–16 years of age. These molecular results provide a basis for a database essential for the taxonomic framework, cultivar development and germplasm conservation of eucheumatoids

Universal antenatal human immunodeficiency virus testing in Hong Kong: consensus statement.

Following the recommendations of the Advisory Council on AIDS, Hong Kong, the Hospital Authority announced plans to introduce universal antenatal screening for human immunodeficiency virus infection and hence, a consensus conference was held to discuss strategies for implementing such screening in Hong Kong. This paper reports the discussions of the consensus conference. The consensus meeting group consisted of 15 clinicians and scientists from Hong Kong, Macau, and Thailand. Seven commonly asked questions concerning mother-to-child transmission of human immunodeficiency virus were selected for discussion by the participating panellists. Information on the laboratory diagnosis of human immunodeficiency virus infection and the efficacy of preventive measures in reducing mother-to-child transmission of human immunodeficiency virus were reviewed. Data from local studies was also presented and discussed. The timing, potential problems, and cost issues involved in testing all pregnant women in Hong Kong for human immunodeficiency virus were then considered.published_or_final_versio

MicroRNAs in cardiac arrhythmia: DNA sequence variation of MiR-1 and MiR-133A in long QT syndrome.

Long QT syndrome (LQTS) is a genetic cardiac condition associated with prolonged ventricular repolarization, primarily a result of perturbations in cardiac ion channels, which predisposes individuals to life-threatening arrhythmias. Using DNA screening and sequencing methods, over 700 different LQTS-causing mutations have been identified in 13 genes worldwide. Despite this, the genetic cause of 30-50% of LQTS is presently unknown. MicroRNAs (miRNAs) are small (∼ 22 nucleotides) noncoding RNAs which post-transcriptionally regulate gene expression by binding complementary sequences within messenger RNAs (mRNAs). The human genome encodes over 1800 miRNAs, which target about 60% of human genes. Consequently, miRNAs are likely to regulate many complex processes in the body, indeed aberrant expression of various miRNA species has been implicated in numerous disease states, including cardiovascular diseases. MiR-1 and MiR-133A are the most abundant miRNAs in the heart and have both been reported to regulate cardiac ion channels. We hypothesized that, as a consequence of their role in regulating cardiac ion channels, genetic variation in the genes which encode MiR-1 and MiR-133A might explain some cases of LQTS. Four miRNA genes (miR-1-1, miR-1-2, miR-133a-1 and miR-133a-2), which encode MiR-1 and MiR-133A, were sequenced in 125 LQTS probands. No genetic variants were identified in miR-1-1 or miR-133a-1; but in miR-1-2 we identified a single substitution (n.100A> G) and in miR-133a-2 we identified two substitutions (n.-19G> A and n.98C> T). None of the variants affect the mature miRNA products. Our findings indicate that sequence variants of miR-1-1, miR-1-2, miR-133a-1 and miR-133a-2 are not a cause of LQTS in this cohort

Evaluative and discriminative properties of the Portuguese MacNew Heart Disease Health‐related Quality of Life Questionnaire.

Qual Life Res. 2005 Dec;14(10):2335-41. Evaluative and discriminative properties of the Portuguese MacNew Heart Disease Health-related Quality of Life Questionnaire. Leal A, Paiva C, Höfer S, Amado J, Gomes L, Oldridge N. Cardiac Rehabilitation Unit, Hospital Sto António Porto, Largo Prof Abel Salazar 400, Portugal. [email protected] Abstract The aim of this study was to validate the Portuguese version of the self-administered MacNew Heart Disease Health-related Quality of Life (MacNew) questionnaire in patients after diagnosis of acute coronary syndrome. The MacNew, with a Global score and physical, emotional and social subscales, the Short Form SF-36 (SF-36) and the Hospital Anxiety and Depression Scale (HADS) were completed at baseline by 150 patients and again by 48 clinically stable patients 2-3 weeks later. A cohort of 50 different patients completed the same questionnaires before and after a cardiac rehabilitation program in order to examine responsiveness. Acceptance of the MacNew by the patients was good and the three factor model was substantiated and explained 52.2% of the variance. Internal consistency, intra-class-correlation, and test-retest reliability each exceeded 0.72. The predicted construct validity hypotheses were partially confirmed. The discriminative validity of the MacNew was confirmed with significantly higher MacNew scores for patients with normal left ventricular function, with improved health status, and who were not anxious or depressed. Even though MacNew scores improved significantly following cardiac rehabilitation, the evaluative validity of the MacNew was less robust with small responsiveness statistics. The Portuguese version of the MacNew HRQL questionnaire appears to be a reliable, valid, and moderately responsive instrument to evaluate health-related quality of life after diagnosis of acute coronary syndrome. PMID: 16328913 [PubMed - indexed for MEDLINE

Decline in Clostridium difficile-associated disease rates in Singapore public hospitals, 2006 to 2008

<p>Abstract</p> <p>Background</p> <p><it>Clostridium difficile </it>is the major cause of pseudomembranous colitis associated with antibiotic use, and the spread of the hypervirulent epidemic ribotype 027/NAP-1 strain across hospitals worldwide has re-focused attention on this nosocomial pathogen. The overall incidence and trend of <it>C. difficile</it>-associated disease (CDAD) in Singapore is unknown, and a surveillance program to determine these via formal laboratory-based reporting was established.</p> <p>Findings</p> <p>Laboratory and pharmacy data were collated from one tertiary and two secondary hospitals on a quarterly basis between 2006 and 2008. All hospitals tested for <it>C. difficile </it>using Immunocard Toxins A&B (Meridian Bioscience Inc., Cincinnati, OH) during this period. Duplicate positive <it>C. difficile </it>results within a 14-day period were removed. The CDAD results were compared with trends in hospital-based prescription of major classes of antibiotics.</p> <p>Overall CDAD incidence-density decreased from 5.16 (95%CI: 4.73 - 5.62) cases per 10,000 inpatient-days in 2006 to 2.99 (95%CI: 2.67 to 3.33) cases per 10,000 inpatient-days in 2008 (<it>p </it>< 0.001), while overall rates for <it>C. difficile </it>testing increased significantly (<it>p </it>< 0.001) within the same period. These trends were mirrored at the individual hospital level. Evaluation of antibiotic prescription data at all hospitals showed increasing use of carbapenems and fluoroquinolones, while cephalosporin and clindamycin prescription remained stable.</p> <p>Conclusions</p> <p>Our results demonstrate a real decline of CDAD rates in three large local hospitals. The cause is unclear and is not associated with improved infection control measures or reduction in antibiotic prescription. Lack of <it>C. difficile </it>stool cultures as part of routine testing precluded determination of the decline of a major clone as a potential explanation. For more accurate epidemiological trending of CDAD and early detection of epidemic clones, data collection will have to be expanded and resources set in place for reference laboratory culture and typing.</p