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Biomedical subjects

H C Lim

Publications and source records attributed to H C Lim.

At least 19 recordsLinked to original sources
PubMed1995

Temperature induction of bacteriophage lambda mutants in Escherichia coli.

The paper presents temperature induction in Escherichia coli cells with phage lambda on the target-protein production and cell growth. Replicated lambda-DNA particles in the Q- and S- mutants remain naked for a longer time by preventing DNA packaging and cell lysis, and therefore the expression of the foreign genes is high. However, the parasitic infection of phage-lambda causes on significant losses of host cell viability in the induction phase. The temperature effects on cell growth and targeted-gene product formation were investigated. Gene amplification was found to be growth phase dependent for both Qam73 (Q mutation) and Sam100 (S mutation) mutants. Maximum induction occurs in the early exponential phase and under the optimal cell density. The total beta-galactosidase activity at this optimal induction condition increases roughly 8-10-fold with respect to that without thermal induction. To maximize the induction efficiency for the gene-product beta-galactosidase activity, several operating parameters were investigated. In this study, temperature induction is strongly dependent upon the population density of 'susceptible' cells at which time the temperature is shifted to 38-42 degrees C. This may be due to the 'threshold' population density to regulate the infection of lambda to hosts and control the productivity of target gene expression.

Bacteriophage lambda↗
PubMed1994

cAMP, ethanol, and CO2 production with the addition of D-glucose anomer to starved yeast cells.

cAMP, ethanol, and CO2 production in starved yeast cells after the addition of D-glucose anomer was measured and compared over a wide range of anomer concentrations. At 1.0 g/l or higher concentrations, the addition of beta-D-glucose resulted in a higher cAMP peak. beta-D-glucose was more rapidly metabolized to ethanol and CO2 than alpha-D-glucose, although there was no notable difference in the uptake rates of the two anomers. At 0.4 g/l D-glucose anomer, the differences in cAMP and ethanol production rates for the two anomers were not significant. At 0.2 g/l D-glucose anomer or lower concentrations, ethanol production with alpha-D-glucose was higher than that with beta-D-glucose. The uptake rate of alpha-D-glucose was higher than that of beta-D-glucose at this low concentration.

Carbon Dioxide↗
PubMed1994

Tyrosinase production in recombinant E. coli containing trp promoter and ubiquitin sequence.

We have successfully expressed the active tyrosinase of Streptomyces antibioticus in Escherichia coli under the control of the trp promoter by fusing the sequence to the ORF438 gene. Because our attempt to connect the polycistronic gene of ORF438 and tyrosinase directly to the trp promoter of E. coli resulted in the expression of functionally inactive tyrosinase, we decided to fuse the COOH-terminus of ubiquitin sequence to the NH2-terminus of ORF438. Ubiquitin fusion has been shown to augment the yield of cloned gene products in E. coli by increasing the stability or translational efficiency of the fusion proteins. As a result, E. coli transformants harboring a plasmid pTRUBF that contains the ubiquitin-fused ORF438 and the tyrosinase gene produced the strong black pigment of melanin. About 300 units of tyrosinase per liter of batch culture were detected when cultivated in M9 medium containing casamino acids, L-tyrosine, and copper supplements. The black pigment, however, was not seen when grown in LB medium, suggesting that the trp promoter is well regulated. When recombinant E. coli cells grown in LB medium were transferred to a tryptophan-deficient minimal medium with phenol, we observed that phenol was removed from the solution, and the color of the medium turned black. This is due to the fact that the tyrosinase has polyphenol oxidase properties. We expect to use this recombinant E. coli for the waste treatment of phenolic compounds.

Base Sequence↗
PubMed1994

Standardisation and adaptation of the Denver Developmental Screening Test (DDST) and Denver II for use in Singapore children.

OBJECTIVE: To modify and standardise the Denver Developmental Screening Test (DDST) and Denver II for developmental screening of children in Singapore. METHOD: The study used a quota sample of 2,194 Singapore children aged 4 weeks to 6 years. Logistic regression analysis established the 25th, 50th, 75th, and 90th percentile passing age for achieving the test tasks. Subgroup differences in Sex, Ethnicity, Social Class and Mother's Education were analysed by stepwise logistic regression; the composite norms for items with statistically significant subgroup differences (p < or = 0.10), were then adjusted by weighting based on the composition of Singapore children. The study protocol was based on the DDST (1975) and Denver II (1990), the latest version of the DDST. Modifications were introduced to improve on the sensitivity of the test and to make the test more suited to Singapore culture. MAIN FINDINGS: Out of the 215 items studied, 115 items were selected to form the new test, DDST, Singapore, DDST, Singapore shares 63% of the items with DDST (1975) and 67% of the items with Denver II. Among the comparable items, differences between the norms of Singapore and Denver children greater than 10% were demonstrated in more than 30 items, and differences of greater than 20% in 10 items. Within the study sample of Singapore children, there were relatively smaller differences among the subgroups studied. Only 10 items had clinically significant subgroup differences of more than 10%. None had more than 20% difference. CONCLUSION: DDST, Singapore is substantially different from the DDST (1975) and Denver II (1990). The use of the local standardised version for developmental screening of Singapore children is justified.

Child↗
PubMed1992

Nitrazepam-induced cricopharyngeal dysphagia, abnormal esophageal peristalsis and associated bronchospasm: probable cause of nitrazepam-related sudden death.

Nitrazepam was used in the treatment of resistant myoclonic epilepsy in 38 children. After the occurrence of nitrazepam-associated swallowing incoordination, high-peaked esophageal peristalsis and related bronchospasm in one patient, we initiated a prospective study of esophageal manometry using a station pull-through technique with a pediatric 4-channel continuous perfusing system. Three more patients were found to have delayed cricopharyngeal relaxation and high-peaked esophageal peristaltic waves. The initial patient developed severe respiratory distress and bronchospasm necessitating ventilatory support while on nitrazepam and improved dramatically with subsequent normal manometric study following nitrazepam discontinuation. Nitrazepam was reintroduced for its anticonvulsant and cognitive benefits and was tolerated at a reduced dosage. We postulate a central nervous system effect of nitrazepam promoting parasympathetic overactivity or vagotonia which can cause potentially fatal respiratory distress. Care must be exercised in nitrazepam use and esophageal manometry may be helpful in defining patients at greater risk for sudden death.

Adolescent↗
PubMed1988

Concomitant hypotensive and antinociceptive effects of guanabenz in conscious rats: involvement of nucleus reticularis gigantocellularis.

Subcutaneous administration of guanabenz (1, 2, 5, or 8 mg/kg) in conscious Sprague-Dawley rats exerted concomitant suppressive actions on systolic pressure (tail-cuff sphygmomanometric measurement) and pain responses (hot-plate algesio-metric assay) that varied both in degree (antinociception much greater than hypotension) and response pattern. Furthermore, both depressive effects were appreciably attenuated by bilateral lesions of the nucleus reticularis gigantocellularis in the medulla oblongata. It is speculated that separate subpopulations of neurons within this reticular nucleus may be involved in the expression of hypotension and antinociception by this amino-guanidine derivative.

Analgesia↗
PubMed1988

Characterization of alpha-adrenoceptors in the nucleus reticularis gigantocellularis involved in the cardiovascular depressant effects of guanabenz in the rat.

The participation of alpha-adrenoceptors in the nucleus reticularis gigantocellularis in the hypotensive, negative inotropic and chronotropic effects induced by guanabenz, was examined in rats anesthetized with pentobarbital sodium (40 mg/kg, i.p.). Pretreatment with alpha-adrenoceptor antagonists yohimbine (10 micrograms), phentolamine (2.5 micrograms) and phenoxybenzamine (20 micrograms), which were injected bilaterally into the nucleus reticularis gigantocellularis, significantly antagonized the cardiovascular suppressant effects normally produced by systemic administration of guanabenz (10 micrograms/kg, i.v.). Pretreatment with prazosin (0.25 microgram) did not affect the vasodepressive, but significantly attenuated the bradycardic actions of guanabenz. The general trend of "antagonization potency" shown by the alpha-adrenergic blockers, against the cardiovascular effects of guanabenz, was in the order: yohimbine greater than phentolamine greater than phenoxybenzamine greater than prazosin. It is concluded that while the alpha 2-adrenoceptors in the nucleus reticularis gigantocellularis are more critically involved in the antihypertensive actions of guanabenz, the possibility exists that alpha 1-adrenoceptors may also participate, in part.

Adrenergic alpha-Antagonists↗
PubMed1986

The roles of alpha 2-adrenoceptors in the nucleus reticularis gigantocellularis and vagal mechanism in the cardiovascular suppressive effects of guanabenz in the rat.

In pentobarbital-anesthetized rats, pretreatment with yohimbine (10 micrograms), which was microinjected into the bilateral nucleus reticularis gigantocellularis (NRGC), significantly antagonized the reduction in arterial pressure, and the force and rate of heart contraction normally promoted by systemic administration of guanabenz (10 micrograms/kg, i.v.). At the same time, the vasodepressive as well as negative inotropic and chronotropic effects of direct application of guanabenz (500 ng) into the NRGC were attenuated by bilateral cervical vagotomy or atropine sulfate (1 mg/kg, i.v.). We conclude that guanabenz may promote antihypertension by activating the alpha 2-adrenoceptors in the NRGC, which in turn elicits cardiovascular suppression by at least facilitating the vagal outflows to the heart.

Animals↗
PubMed1985

Pulmonary edema as a cause for fulminant death following microinjection of kainic acid into the rat medulla oblongata.

The present study confirmed histopathologically the suggestion that pulmonary edema may underlie the fulminant death following microinjection of kainic acid into many medullary sites in rats anesthetized with pentobarbital sodium. We also observed a sustained elevation in systemic arterial pressure, cardiac contractility and heart rate before the animal succumbed to kainate neurotoxicity. It is suggested that the increased systemic hydrostatic pressure may retrogradely promote pulmonary hypertension that precipitates pulmonary edema.

Animals↗
PubMed1985

Participation of nucleus reticularis gigantocellularis in guanabenz-promoted hypotension, decrease in cardiac contractility and bradycardia in rats.

The possible participation of the nucleus reticularis gigantocellularis in the hypotension, decrease in cardiac contractility and bradycardia induced by guanabenz was examined in rats that were anesthetized with pentobarbital sodium (40 mg/kg, i.p.). Guanabenz (10 micrograms/kg, i.v.) elicited an initial, transient hypertension, accompanied by an increase in cardiac contractility, followed by a significant and sustained hypotension, as well as decrease in the force and rate of cardiac contraction. In rats receiving bilateral focal electrolytic lesions of the nucleus reticularis gigantocellularis, the same injection produced only the initial transient responses, without the subsequent depressant effects. Microinjection of guanabenz directly into the ventro-medial portion of the nucleus reticularis gigantocellularis, at an ineffective systemic concentration (500 ng), produced significant and prolonged reduction in arterial pressure, cardiac contractility and heart rhythm. On the other hand, local application of the same concentration (500 ng) of guanabenz into the lateral portion of the same nucleus produced only minor hypotension and bradycardia, with no decrease in cardiac contractility. It is concluded that the nucleus reticularis gigantocellularis is at least one of the central sites through which guanabenz may produce its cardiovascular suppressant effects.

Animals↗
PubMed1984

Iron status in blood donors.

The iron status of 592 blood donors was analysed. The reduction in iron stores was related to the frequency rather than the total number of donations. 8% had reduced iron stores but this was not related to blood donation, iron supplementation and sex of the donors. Serum ferritin rather than haemoglobin, serum iron and transferrin saturation is the most reliable indication of iron status of blood donors. Iron supplementation did not confer any benefits.

Blood Donors↗
PubMed1978

Formaldehyde incorporation by a new methylotroph (L3).

A number of bacterial strains have been isolated and investigated in our search for a promising organism in the production of single-cell protein from methanol. Strain L3 among these isolates was identified as an obligate methylotroph which grew only on methanol and formaldehyde as the sole sources of carbon and energy. The organism also grew well in batch and chemostat mixed-substrate cultures containing methanol, formaldehyde, and formate. Although formate was not utilized as a sole carbon and energy source, it was readily taken up and oxidized by either formaldehyde- or methanol-grown cells. The organism incorporated carbon by means of the ribulose monophosphate pathway when growing on either methanol, formaldehyde, or various mixtures of C1 compounds. Its C1-oxidation enzymes included phenazine methosulfate-linked methanol and formaldehyde dehydrogenase and a nicotinamide adenine dinucleotide-linked formate dehydrogenase. Identical inhibition by formaldehyde of the first two dehydrogenases suggested that they are actually the same enzyme. The organism had a rapid growth rate, a high cell yield in the chemostat, a high protein content, and a favorable amino acid distribution for use as a source of single-cell protein. Of special interest was the ability of the organism to utilize formaldehyde via the ribulose monophosphate cycle.

Alcohol Oxidoreductases↗
PubMed1977

An analysis of extended and exponentially-fed-batch cultures.

Under certain conditions it is shown that an extended culture is equivalent to an exponentially-fed-batch culture, that an exponentially-fed-batch culture (and an extended culture) can be maintained at a steady state and that an exponentially-fed-batch culture may be mimicked by a continuous-flow culture with a constant dilution rate. Operational conditions required to maintain steady states are specified.

Bacteria↗
PubMed1977

Growth characteristics of a new methylomonad.

A methylomonad culture was isolated from pond water and examined as a potential source of single-cell protein. A medium containing magnesium sulfate, ammonium hydroxide, sodium phosphate, tap water, and methanol supported the growth of the isolate. Optimal growth conditions in batch cultures for the organism were: temperature, 30 to 33 degrees C; pH 7.1; and phosphate concentration, 0.015 M. The minimum doubling time obtained was 1.6 h. The specific growth rate in batch culture was dependent on the methanol concentration, reaching a maximum around 0.2% (wt/vol). Growth inhibition was apparent above 0.3% (wt/vol), and growth was completely inhibited above 4.6% (wt/vol) methanol. Although the inhibitory effect of formaldehyde on the specific growth rate was much greater than that of formate, the organism utilized formaldehyde, but not formate, as a sole carbon and energy source in batch cultures. The isolate was identified primarily by its inability to utilize any carbon source other than methanol and formaldehyde for growth. Although it is capable of rapid growth on methanol, the organism showed a very weak catalase activity. The amino acid content of the cells compared favorably with the reference levels for the essential amino acids specific by the Food and Agricultural Organization of the United Nations.

Amino Acids↗
PubMed1976

A model for enzymatic isomerization of D-glucose to D-fructose in a batch reactor.

Using whole cells containing glucose isomerase, mathematical models for the enzymatic conversion of D-glucose to D-fructose and for the inactivation of the enzyme catalyst have been postulated and verified experimentally. The heat of reaction, the equilibrium constant, and the individual rate constants and their activation energies have been estimated. The model can be used to predict the time course for the enzymatic production of fructose in a batch reactor within the tested experimental range of 40-80 degrees C.

Cells↗