Rapid vinculin exchange dynamics at focal adhesions in primary osteoblasts following shear flow stimulation.

The adaptor protein vinculin plays a key position in the formation of focal adhesions and regulates cell attachment. To study the turnover of vinculin in bone-derived cells, we expressed green fluorescent protein-tagged vinculin in primary bovine osteoblasts and examined the appearance of focal adhesions in cells exposed to laminar shear flow. Already 20 sec after application of shear stress fluorescently labelled focal adhesions became visible as small flashing dots at the periphery of cells. The number of newly formed focal adhesions per individual cell increased continuously over approximately 300 sec and then remained relatively stable. The assembly of focal adhesions in shear stress-stimulated osteoblasts was accompanied by a transient rise in intracellular calcium levels. The mean assembly time of an individual focal adhesion plaque was 32.2+/-2.2 sec and the mean disassembly time was 60.5+/-6.0 sec. The recruitment of vinculin to nascent focal adhesions was in the same range as the recovery half-life of GFP-vinculin at stable focal adhesions (13.0+/-2.0 sec). These data show that accumulation of GFP-vinculin in newly formed focal adhesions and its exchange from pre-existing, mature plaques are both rapid processes that occur in mechanically stimulated osteoblasts within less than one minute.

ANTIMIC: a database of antimicrobial sequences.

Antimicrobial peptides (AMPs) are important components of the innate immune system of many species. These peptides are found in eukaryotes, including mammals, amphibians, insects and plants, as well as in prokaryotes. Other than having pathogen-lytic properties, these peptides have other activities like antitumor activity, mitogen activity, or they may act as signaling molecules. Their short length, fast and efficient action against microbes and low toxicity to mammals have made them potential candidates as peptide drugs. In many cases they are effective against pathogens that are resistant to conventional antibiotics. They can serve as natural templates for the design of novel antimicrobial drugs. Although there are vast amounts of data on natural AMPs, they are not available through one central resource. We have developed a comprehensive database (ANTIMIC, http://research.i2r. a-star.edu.sg/Templar/DB/ANTIMIC/) of known and putative AMPs, which contains approximately 1700 of these peptides. The database is integrated with tools to facilitate efficient extraction of data and their analysis at molecular level, as well as search for new AMPs. These tools include BLAST, PDB structure viewer and the Antimic profile module.

Effects of solid retention time on the performance of submerged anoxic/oxic membrane bioreactor.

In this study, four similar bench-scale submerged Anoxic/Oxic Membrane Bioreactors (MBR) were used simultaneously to investigate the effects of solids retention time (SRT) on organic and nitrogen removal in MBR for treating domestic wastewater. COD removal efficiencies in all reactors were consistently above 94% under steady state conditions. Complete conversion of NH(4+)-N to NO(3-)-N was readily achieved over a feed NH(4+)-N concentration range of 30 to 50 mg/L. It was also observed that SRT did not significantly affect the nitrification in the MBR systems investigated. The average denitrification efficiencies for the 3, 5, 10 and 20 days SRT operations were 43.9, 32.6, 47.5 and 66.5%, respectively. In general, the average effluent nitrogen concentrations, which were mainly nitrate, were about 22.2, 27.6, 21.7 and 13.9 mg/L for the 3, 5, 10 and 20 days SRT systems, respectively. The rate of membrane fouling at 3 days SRT operation was more rapid than that observed at 5 days SRT. No fouling was noted in the 10 days and 20 days SRT systems during the entire period of study.

Cloning, characterization and expression of a cDNA clone encoding rabbit ubiquitin-conjugating enzyme, E2(32k).

A cDNA clone encoding rabbit E2(32k) was obtained by library screening and PCR. The cDNA contains an open reading frame coding for 238 amino acids which shows an overall identity of 81% to human CDC34, the cell cycle-related ubiquitin-conjugating enzyme. A 50% homology to yeast CDC34 within the conserved core domain was also observed. Northern blot analysis indicated that three transcripts existed in all six rabbit tissues examined but their expression levels varied over a wide range. The putative cDNA coding region was highly expressed in Escherichia coli as a his-tagged protein which was purified to homogeneity. The ability of this expressed protein to form a thiolester bond with ubiquitin showed that it was functionally active. The ability of this protein to catalyze the conjugation of ubiquitin to histone H2A and H2B was also examined.

A functional significance for codon third bases.

Most amino acids are specified by more than one trinucleotide codon. Here we show that amino acids of differing functional importance may be distinguished by the pattern of synonymous codon usage. GC-rich genes tend to be of a greater transcriptional (p<0.01) and mitogenic (p<0.0001) significance than AT-rich genes, consistent with GC-->AT mutational drift in methylated genomic regions. Third-base GC retention also identifies critical amino acids within individual proteins, as indicated by non-random patterns of codon variation between gene homologs and also by differential sequelae of site-directed mutagenesis. Sequence analysis of human receptor tyrosine kinase genes confirms that functionally important transmembrane hydrophobic amino acids are specified by codons containing GC third bases more often than are transmembrane neutral amino acids (chi(2)=134.2). Amino acids encoded by GC third bases thus appear more tightly linked to cell function and survival than are those encoded by AT third bases.

Sequence analysis of the major outer membrane protein gene of an ovine abortion strain of Chlamydia psittaci.

The major outer membrane protein (MOMP) gene from an ovine abortion strain of Chlamydia psittaci (S26/3) has been cloned and sequenced. The gene shows the features of other chlamydial MOMPs but comparison with the previously reported sequence for the ovine abortion isolate A22/M has revealed substantial sequence divergence which is clustered into the same four intramolecular regions as the sequence variation found between C. trachomatis serovars. Subsequent restriction enzyme analysis of A22/M DNA has shown that it has an avian-type genomic profile and thus the comparison is between types rather than between strains.

Antibody response of the ovine lymph node to experimental infection with an ovine abortion strain of Chlamydia psittaci.

After primary infection with Chlamydia psittaci in the draining area of the popliteal lymph node, viable organisms could be isolated from the efferent lymph only before the primary immune response developed. The lymph antibody response, as assayed by the complement fixation and immunofluorescence (IF) antibody tests, showed a rise in titre that peaked approximately 2 weeks after infection. Immunoblotting revealed that antibodies produced during this period were predominantly directed against the major outer membrane protein (MOMP). In secondary infection of convalescent sheep, an elevated IF antibody titre, already present in the lymph and blood, could not be boosted. Viable organisms could not be isolated from these sheep. Antibodies produced reacted to 12-14 bands in the immunoblot profile including the MOMP band. These potentially immunoprotective antigens, particularly MOMP, should be considered as useful candidates for an improved vaccine against ovine enzootic abortion, in further investigations.

Efficacy against ovine enzootic abortion of an experimental vaccine containing purified elementary bodies of Chlamydia psittaci.

A vaccine prepared from purified, inactivated elementary bodies of Chlamydia psittaci protected sheep against abortion after subcutaneous challenge with live chlamydiae. Immunoblot analysis of serum samples revealed a consistently dominant antibody response against the chlamydial major outer membrane protein in all vaccinated sheep. Reactions to other chlamydial antigens were also detected but were less pronounced or inconsistent. Serological responses detected by complement fixation were variable and did not correlate with immunity.

Curcumin inhibited the arylamines N-acetyltransferase activity, gene expression and DNA adduct formation in human lung cancer cells (A549).

It is well known that N-acetyltransferase (NAT) plays an important role in the arylamine metabolism. We analysed the response of A549 human lung cancer cells for N-acetylation of 2-aminofluorene (AF) to curcumin. After curcumin treatment, the NAT activity was examined by HPLC, AF-DNA adduct formation was examined by HPLC, and NAT gene expression by polymerase chain reaction were detected. The NAT activity in the human A549 cells and cytosols was suppressed by curcumin in a dose-dependent manner. The results also demonstrated that gene expression (NAT1 mRNA) in human lung A549 tumor cells was inhibited and decreased by curcumin. After the incubation of human lung A549 tumor cells with AF with or without curcumin co-treatment, the cells were recovered and DNA was prepared and hydrolyzed to nucleotides. The adducted nucleotides were extracted into butanol and analyzation of AF-DNA adducts was done by HPLC. The results also demonstrated that curcumin decreases AF-DNA adduct formation in the human lung A549 tumor cells.

The role of gastric muscle relaxation in cytoprotection induced by san-huang-xie-xin-tang in rats.

We examined the mechanism of the protective effect of San-huang-xie-xin-tang (SHXT), a traditional oriental drug, on the gastric mucosa. SHXT, given intraduodenally, inhibited the gastric secretion in pylorus-ligated rats. SHXT, given orally 30 min before the administration of aspirin, HCl-aspirin and ethanol, protected the gastric mucosa from these agent-induced gastric lesions, although it had no effect on the gastric lesions induced by indomethacin and water immersion stress. Oral administration of SHXT increased gastric contents (muscle relaxation) when they were measured 30 min after SHXT administration. SHXT also inhibited carbachol-contracted gastric muscle in vitro. Neither increased gastric contents nor inhibited ethanol lesions of SHXT were observed when these were tested 4 h after SHXT administration. Pretreatment with indomethacin which is a prostaglandin synthesis inhibitor, and N-ethylmaleimide which is a sulfhydryl blocker did not influence either the increased gastric contents or inhibited ethanol lesions of SHXT. These results indicate that gastric muscle relaxation plays an important role in the gastric protective mechanisms of SHXT, and the endogenous prostaglandins and the sulfhydryl compounds are not necessary for the action of SHXT.

The effects of vandellia cordifolia on renal functions and arterial blood pressure.

Vandellia cordifolia (COLSM) G. DON of Scrophulariaceae (V. cordifolia) is an annual wild herb indigenous to Taiwan. It can be found in plains, low altitudes, swampy places, and paddy fields. Taiwanese folk physicians use it in "nephritis, uremia, furnucle, carbuncle." The LD50 (95% confidence limit) of the crude extract of V. codifolia given by the oral route was more than 10 g/kg in rats. By the intraperitoneal route, it was 4.6 g/kg (4.35-4.93). The extraction rate was 16.6%. We studied its effects on renal functions and blood pressure and found that (1) it had diuretic effect on normal rats, (2) it decreased glomerular filtration rate and renal blood flow on normal kidneys in rabbits, (3) it had no effects on glomerular filtration rate and renal blood flow on glycerin-induced insufficient kidneys in rabbits, (4) it had diuretic effects on both normal and glycerin-induced insufficient kidneys in rabbits, (5) it could inhibit Na+ and K+ reabsorption on normal and glycerin-induced insufficient kidneys in rabbits, (6) it had hypertensive effect and this effect could be blocked by phenoxybenzamine. From the above facts, we conclude that V. cordifolia has diuretic effect and it may act on renal tubules to inhibit Na+ and K+ reabsorption.

Testicular seminoma--the results of treatment by radiotherapy in Singapore.

A retrospective analysis was done of 49 patients treated for testicular seminoma over a 15-year period between January 1980 and April 1995 at the Department of Therapeutic Radiology, Singapore General Hospital. Their mean age was 39 years and their median period of follow-up was 67 months. Using the Royal Marsden Staging classification, 35 (71.4%) were stage I, 10 (20.4%) were stage II, I (2%) was stage III, and 3 (6.1%) were stage IV. Five (10.2%) arose in undescended testes. Thirty-six patients were treated with inguinal orchidectomy and postoperative radiotherapy to the para-aortic and ipsilateral iliac nodes. Eight had para-aortic and bilateral iliac irradiation, and 2 had ipsilateral pelvic, para-aortic, mediastinal and supraclavicular lymph node irradiation. Six of the patients had, in addition, irradiation of the inguinal-scrotal area. Nine received both chemotherapy as well as radiotherapy. The median dose of radiation given was 30.67 Gy and where a boost was required, the median total tumour dose was 35.75 Gy. The actuarial relapse-free survival at 10 years was 88.9% and the actuarial overall survival at 10 years was also 88.9%. Long-term treatment toxicities were uncommon.

Localised nodal non-Hodgkin's lymphoma--a study of 42 cases treated by radiotherapy.

The records of 42 patients with stage I and II nodal non-Hodgkin's lymphoma treated by radiotherapy with and without chemotherapy were analysed with regards to clinical presentation, investigations, treatment, radiotherapy techniques and results. The 5-year overall survival rates for all, low grade, intermediate grade and high grade histology types are 69.5%, 100%, 64.4% and 46.9% respectively. The 5-year relapse-free survival rates of those treated by initial radiotherapy alone for all, low grade and intermediate-high grade histology types are 54.8%, 46.9% and 59.4% respectively.

Patient profile and survival in 270 computer tomography-staged patients with nasopharyngeal cancer treated at the Singapore General Hospital.

This is a report analysing the patient profile and survival results in 270 patients with nasopharyngeal carcinoma treated with radiation therapy between July 1987 and December 1988. The patients had a median age of 52.6 months, and a male-female ratio of 2.5:1. Ninety-four percent of patients were Chinese. The commonest presentation was that of a neck mass (found in approximately 65% of patients). Approximately two-thirds presented with disease more advanced than Ho's stage II. At a median follow-up of 52.6 months, the 5-year actuarial survival rate was 53% and the freedom from relapse rate was 57%. One hundred and seven patients died within the observation period. Of these, 46 patients had disease in the post nasal space, 41 in the neck and 60 in sites outside the head and neck.

Removal of MS2 bacteriophage using membrane technologies.

Removals of MS2 bacteriophage virus using different membrane materials under different operating pressures were investigated. The results obtained in this study suggested that a better log removal in terms of MS2 bacteriophage virus could be achieved using Polyamide RO membrane under the optimum operating pressure of 100 psi. It is further noted that variable MS2 influent concentration levels resulted in corresponding variable log removals of the bacteriophages by the Polyamide RO membrane. The presence of MS2 bacteriophage virus in the effluent could possibly be due to leakage of bacteriophages through the membranes structure. Investigations using SEM and AFM showed that there were gaps or pores present in the membrane structure which were sufficiently large for the MS2 viruses to pass through.

Integration of bioInformatics tools at the National University of Singapore (NUS).

In the past decade "Big Science" such as the Genome Project has generated an enormous amount of data in the life sciences. Concurrently, the synergy of this project with existing research has quickened the pace of biological discovery. But the major drawback that is beginning to be felt worldwide is the primitive level of organisation in the data accumulated. Without a proper framework or knowledge scaffold to hang and interconnect the various bits of data and information, the national knowledge-to-data ratio is declining rapidly. We are trying to serve a solution to this enigma by providing a World Wide Web (WWW) interface to Biosoftware and at the same time have come up with a database integration tool that can query heterogeneous, geographically scattered and disparate databases simultaneously. In this report we will talk about BioInformatics in general with specific reference to BioInformatics Centre (BIC) at the National University of Singapore.

Influence of Chinese cultural health beliefs among Malaysian Chinese in a suburban population: a survey.

INTRODUCTION: In a multiethnic nation, it is not uncommon for doctors to encounter patients of different cultural backgrounds. Often, patients' cultural beliefs influence their perception of health and illnesses, and their treatment option. Many Chinese cultural beliefs are influenced by the Taoist concept of yin-yang balance. METHODS: We interviewed 50 Malaysian Chinese from the general public of a suburban population in order to unravel the impact of Chinese cultural health beliefs on their decision-making, and this was compared with the opinions of 50 Chinese medical students from second to final year. Convenience sampling was then applied. RESULTS: From the survey, 78 percent of the general public believed that 'too much heat' or 'too much coldness' in the body could cause diseases. Compared to the medical students, a significantly higher number of the respondents held such beliefs, including the beliefs that abdominal colic is due to excessive 'wind' in the abdomen, consuming certain food can dispel wind from the body, and the importance of observing taboos during the confinement period after delivery. The majority of respondents from both groups believed that it is acceptable to combine both traditional Chinese medicine and modern medicine. CONCLUSION: There is a discrepancy in the extent to which these beliefs influence the perception of health and illnesses among the general public and among medical students. Healthcare providers need to be aware of such beliefs and practices regarding traditional Chinese medicine among their Chinese patients.

Multi-variate analysis of burns patients in the Singapore General Hospital Burns Centre (2003-2005).

The Burns Centre at the Singapore General Hospital (SGH) serves as a tertiary referral centre for burns management for Singapore's 4 million residents as well as the Southeast Asia region. Our study is a multivariate analysis of all burns patients admitted between 2003 and 2005. A total of 482 patients were admitted during this period with an average annual admission of 161. This represents a low incidence of 0.04 per 1000 admissions for the Singapore population. 13.3% of the study population were children, which is lower than previous studies. The mean age at admission was 35 years old and the male:female ratio was 1.9:1. We found a significant difference in age between the local and foreign patients, with the latter being younger. Our study demonstrated a 7.3% increase in cases of occupational burns. The bulk of our patients (57.3%) were directly admitted from SGH's Accident and Emergency Department. The patient characteristics of the various referral sources were found to be very different. GP referrals had significantly lower TBSA while overseas patients had significantly higher TBSA and longer length of stay. The mean and median time to admission was 3.05 days (+/-6.26) and 0 (0-60) day, respectively and the mean and median time to surgery was 7.33 days (+/-8.18) and 5 (0-22) days, respectively. The most common cause of burns was due to scalding. The mean extent of burn (TBSA) was 13.5% (+/-18.0), with significant correlation with the social background. Length of stay was dependent on the need for surgery. The overall mortality rate in this study population was 4.5%, with inhalation injury the main aetiological factor. In addition, the mean duration of the first surgery that patients undergo was significantly longer than that of the second one. This information will be useful for estimating operation times in the future. Finally, Acinetobacter baumannii was the most common bacteria in wound cultures. There is a need for periodic reviews of wound cultures in burns patients in order to modify the preventive and therapeutic strategies against these bacteria.

Stenting before laparoscopic common bile duct exploration: a helpful strategy.

We describe a 43-year-old man who had a difficult endoscopic retrograde cholangiopancreatography, allowing only placement of a stent without removal of the common bile duct stone. He subsequently underwent laparoscopic cholecystectomy with laparoscopic common bile duct exploration and primary closure after laparoscopic choledochotomy.