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Biomedical subjects

H Loh

Publications and source records attributed to H Loh.

At least 19 recordsLinked to original sources
PubMed2005

Effects of opioid antagonists and morphine in a hippocampal hypoxia/hypoglycemia model.

The influence of opioid antagonists and of morphine on rat hippocampal slices in a model of reversible hypoxia/hypoglycemia was investigated by assessment of evoked field potentials (population spike amplitude). In control slices, a brief hypoxia/hypoglycemia led to a loss of field potentials followed by an impaired recovery (40-50% of baseline) during reperfusion. In contrast, restoration was significantly improved when the opioid receptor antagonists funaltrexamine (mu) or naltrindole (delta) were administered prior to and during hypoxia/hypoglycemia. In addition, recovery was improved in brain slices derived from mu-opioid receptor-deficient mice as compared to wild-type mice, indicating a deleterious role of endogenous opioids in hypoxia/hypoglycemia. Exogenous opiate exposure with morphine (0.1, 1.0, 10 microM) prior to hypoxia/hypoglycemia caused a slight concentration dependent increase of evoked field potentials. When morphine exposure was terminated after 1h and immediately followed by hypoxia/hypoglycemia, an impaired recovery of population spike amplitude was obtained, dependent on morphine concentration during preincubation. These results demonstrate that morphine aggravates neurotoxic effects of hypoxia/hypoglycemia. Conversely, when onset of hypoxia/hypoglycemia was delayed for 3h after morphine termination, a significantly improved recovery was observed. Similarly, in vivo administration of morphine 12h prior to slice preparation resulted in a dose dependent improved recovery of field potentials after hypoxia/hypoglycemia. These results provide evidence that preconditioning with morphine is able to induce neuroprotective effects.

Analgesics, Opioid↗
PubMed2004

Accelerated kindling development in mu-opioid receptor deficient mice.

The relevance of mu-opioid systems for central excitability and kindling related disturbed learning performance was underlined by investigations using mu-opioid receptor knockout mice. Mice lacking mu-opioid receptors showed an accelerated kindling development induced by the convulsant drug pentylenetetrazol. Blockade of delta-opioid receptors by naltrindole suppressing kindling development in wild-type animals led to a further acceleration of kindled seizure development in the knockout mice. Mice lacking mu-opioid receptors showed such a low learning performance in the shuttle box, that the kindling induced learning deficit as seen in wild-type mice was not detected. The results were discussed on the basis of receptor binding studies with regard to subtypes of glutamatergic receptors, delta-opioid and somatostatin receptors. An increase in glutamate and somatostatin binding could contribute to the enhanced excitability in the-mu-opioid receptor knockout mice.

Animals↗
PubMed2004

Some current controversies in heart failure (2004-2005).

The disparity between what doctor's believe and the medical facts can be frightening at times. This reflects centuries of training on how to manage patients in the absence of evidence to guide practice. Increasingly disease is becoming susceptible to treatment, requiring a change in the professional approach from the educated, hopeful guess to the application of objective data. Arrogance and certainty, born of ignorance and clutching in desperation at imaginary straws probably still have a place in medicine when conventional treatment has failed, but should play a diminishing role within the medical profession. Epidemiological association is no longer sufficient evidence to conclude that an intervention is effective; with few exceptions, randomised-controlled trials are required. A more successful and objective era of medical practice has arrived. Consequently, a greater degree of scepticism about claims of benefit is also appropriate, which should apply equally to treatments new and old. Of course, no 2 patients are alike, and the evidence-base remains a guide-map on how patients should be treated, rather than a set of rigid rules. Guidelines are guidelines and not infallible, inviolable decrees. In this article, some of the current controversies in the management of heart failure are discussed, ranging from diagnosis, diastolic heart failure, and the role of natriuretic peptides, to the lack of evidence for a clinically-relevant benefit (and therefore possible harm) from aspirin, statins, implantable defibrillators or revascularisation.

Adrenergic beta-Antagonists↗
PubMed2000

Lack of expression of long-term potentiation in the dentate gyrus but not in the CA1 region of the hippocampus of mu-opioid receptor-deficient mice.

The possible involvement of the mu-opioid receptor subtype in mechanisms of long-term potentiation (LTP) of the lateral perforant pathway to the dentate gyrus neurons, as well as of the Schaffer collateral-commissural input of CA1 neurons, was investigated using mu-opioid receptor-deficient mutant mice. In transversal hippocampal slices from mice lacking the mu-opioid receptor (MOR) only a short potentiation in the dentate gyrus after tetanization of the lateral perforant pathway was found. In contrast, the loss of the mu-opioid receptor in the CA1 region did not affect the potentiation of the field potentials induced by tetanization of the Schaffer collaterals. In parallel experiments, the application of 10 microM of the selective MOR-antagonist, funaltrexamine, decreased LTP in the dentate gyrus of wild-type mice but again did not alter the potentiation of the field potentials in the CA1. The loss of MOR-binding in the hippocampus was accompanied by a reduction in D2-binding sites indicating a possible compensatory role of the dopaminergic system. The D1- and glutamate binding was not affected. These observations confirm earlier results with pharmacological blockade of opioid receptors in the dentate gyrus and demonstrate an essential role of MOR activation for the generation of LTP in the dentate gyrus of the mouse but not necessarily in the CA1 region.

Action Potentials↗
PubMed2000

Characterization of a predominant immunogenic outer membrane protein of Riemerella anatipestifer.

The ompA gene, encoding the 42-kDa major antigenic outer membrane protein OmpA of Riemerella anatipestifer, the etiololgical agent of septicemia anserum exsudativa, was cloned and expressed in Escherichia coli. Recombinant OmpA displayed a molecular mass similar to that predicted from the nucleotide sequence of the ompA gene but lower than that observed in total cell lysates of R. anatipestifer. The ompA gene showed a conserved C-terminal region comprising the OmpA-like domain and a variable N-terminal region. This structure is similar to those of the analogous outer membrane proteins of several gram-negative bacteria. However, OmpA of R. anatipestifer contains six EF-hand calcium-binding domains and two PEST regions, which distinguish it from other outer membrane proteins. The occurrence of these motifs in OmpA suggests a possible role in virulence for this protein. The ompA gene is present in the R. anatipestifer type strain and in all serotype reference strains. However, it exhibits some minor genetic heterogeneity among different serotypes, which seems not to affect the strong antigenic characteristics of the protein. OmpA is a conserved and strong antigenic determinant of R. anatipestifer and hence is suggested to be a valuable protein for the serodetection of R. anatipestifer infections, independent of their serotype.

Amino Acid Sequence↗
PubMed2000

Post-transcriptional regulation of mouse kappa-opioid receptor expression.

Three mRNA variants are generated from the mouse kappa-opioid receptor (KOR) gene. The expression patterns of these KOR mRNA variants in adult animal tissues and during developmental stages are examined. Furthermore, the biological significance of generating these variants is demonstrated with respect to two post-transcriptional mechanisms, i.e., mRNA stability and translation efficiency. Variants A and B are both transcribed from promoter 1 of the KOR gene and expressed from early developmental stages through adult life. Although their sequences differ only at a 30-nucleotide insertion for variant B, these two variants are distinct with regard to their expression patterns, mRNA stability, and translation efficiency. Variant A is expressed ubiquitously in all the tissues examined and has a longer t(1/2) (12 h), whereas variant B is more specific to the central nervous system both pre- and postnatally and has a t(1/2) of approximately 8 h. Variant C is transcribed from promoter 2 of the KOR gene and is most specifically expressed, being detected only in the brain stem, spinal cord, and thalamic/hypothalamic areas of postnatal animals. With regard to protein translation, variants B and C are significantly more efficient than variant A. This study provides the evidence for multiple levels of KOR regulation. The biological implication of the generation of KOR mRNA variants is discussed.

Animals↗
PubMed1999

Molecular fingerprinting of Riemerella anatipestifer by repetitive sequence PCR.

Riemerella anatipestifer is a gram-negative rod-shaped bacterium associated with epizootic infections in poultry. A total of 35 R. anatipestifer isolates including the type strain ATCC11845T, reference and field strains for 18 different serotypes were characterized by repetitive sequence based-PCR (rep-PCR) with outwardly-directed primers based on the repetitive extragenic palindromic (REP) consensus sequence. This technique was applied by using either extracted genomic DNA or preparation of whole bacterial cells harvested directly from plate cultures. Rep-PCR discriminated the R. anatipestifer isolates into 19 electrophoretic types. DNA fingerprints obtained from rep-PCR of extracted genomic DNA or from preparations of whole cells yielded comparable patterns. Substantial variation was seen among the rep-PCR fingerprints of different serotypes. Moreover, different polymorphisms of the rep-PCR fingerprints were evident among epidemiologically unrelated isolates of the same serotype. These results suggest the presence of repetitive extragenic palindromic-like elements within the genome of R. anatipestifer that can be used in some isolates to discriminate between different strains belonging to the same serotype. Rep-PCR may serve as a useful molecular tool for subtyping R. anatipestifer isolates for epidemiologic investigations. The whole cell procedure offers the advantage of ease of performance requiring only small quantities of cells.

Animals↗
PubMed1998

Epidemiological surveillance of melioidosis in Singapore.

During the period 1989 to 1996, a total of 372 cases of melioidosis, with 147 deaths, were reported, giving a mean annual incidence rate of 1.7 per 100,000 population and a case-fatality rate of 39.5%. Majority (89%) of the clinical cases were confirmed by culture of Burkholderia pseudomallei, while the others were presumptive cases based on a single blood specimen with an indirect haemaglutination (IHA) antibody titre of > or = 1:16. The highest incidence rate was reported in those aged 45 years and above (5.7 per 100,000 population), males (2.8 per 100,000 population), and Indian ethnic group (3.0 per 100,000 population). Cases were distributed throughout the island all year round. There was no correlation with rainfall. Most of the cases (77.4%) had other concurrent medical conditions, the most common being diabetes mellitus (57.5%). Factors significantly associated with a higher case-fatality rate were age (55 years and above), septicaemia, smoking history and heart or renal failure. The overall case-fatality rate has been declining from 60% in 1989 to 27% in 1996 due to a greater awareness among medical practitioners to diagnose and treat the disease early. The overall seroprevalence of IHA antibody (titre of > or = 1:16) among asymptomatic population groups was 0.2%. B. pseudomallei isolated from clinical specimens were sensitive to imipenem (100%), ceftazidime (99.1%), piperacillin (99.7%), ampicillin-clavulanate (98.5%), minocycline (97.4%), chloramphenicol (94.3%), doxycycline (94.3%) and tetracycline (93.9%). Of 395 samples of soil collected during epidemiological investigation of reported cases, 1.8% were positive for B. pseudomallei.

Adolescent↗
PubMed1997

Phylogenetic position of Riemerella anatipestifer based on 16S rRNA gene sequences.

Riemerella anatipestifer, the causative agent of septicemia anserum exsudativa (also called new duckling disease), belongs to the family Flavobacteriaceae of gram-negative bacteria. We determined the DNA sequences of the rrs genes encoding the 16S rRNAs of four R. anatipestifer strains by directly sequencing PCR-amplified rrs genes. A sequence similarity analysis confirmed the phylogenetic position of R. anatipestifer in the family Flavobacteriaceae in rRNA superfamily V and allowed fine mapping of R. anatipestifer on a separate rRNA branch comprising the most closely related species, Bergeyella zoohelcum, as well as Chryseobacterium balustinum, Chryseobacterium indologenes, and Chryseobacterium gleum. The sequences of the rrs genes of the four R. anatipestifer strains varied between 0.5 and 1.0%, but all of the strains occupied the same position on the phylogenetic tree. In general, differences in rrs genes were observed among R. anatipestifer strains, even within a given serotype, as shown by restriction fragment length polymorphism of PCR-amplified rrs genes.

Animals↗
PubMed1995

Comparison of Pseudomonas pseudomallei from humans, animals, soil and water by restriction endonuclease analysis.

Pseudomonas pseudomallei isolates from 62 human, 17 animal, 3 soil and 3 water samples were examined by genomic DNA digestion with PstI. Five major (RE I, II, III, IV, V) reproducible restriction patterns were observed, with most (56/62) of the human isolates displaying RE I (30/62), II (5/62), III (15/62), IV (4/62), V (2/62), and the animal (16/17), soil (2/3), water (3/3) isolates showing predominantly RE II profiles. Six human and one soil isolates showed patterns different from those of RE I to V. Restriction endonuclease analysis may be applied in epidemiological studies of melioidosis.

Animals↗
PubMed1990

Separation of platelet concentrates (PC) from buffy coat (BC) using the bottom and top drainage (BAT) system.

By the preparation of platelet concentrates (PC) from buffy coat (BC), the blood component separation may be improved considerably. One can use new bag systems with bottom and top drainage (BAT) which allow partial automation of the separation procedure. Thus, it is possible to produce very pure red cell concentrates (RCC) and FFP, as well as BC, by one separation step under well standardized conditions. Several preparative factors influence the quality of the PCs, especially the storage of the whole blood and the BC. Optimal yields (0.72-0.78 x 10(11) platelets), separation efficiency (67.7-70.8%) and lowest cell contamination (leukocytes 0.1-0.2 x 10(8), red cells 0.2-0.3 x 10(8) per concentrate) were obtained with whole blood stored for 1 h and BC for 2-4 h before resuspension by 30 min face-over-face-rotation. In respect to the release of leukocyte proteases during the storage, the BC should not rest for more than 2-3 h before separation of the platelets in order to prevent potentially functional impairment of the platelets by leukocyte proteases.

Blood Component Removal↗
PubMed1982

Effects of naloxone on d-amphetamine- and apomorphine-induced behavior.

The effects of acute naloxone administration on d-amphetamine- and apomorphine-induced behavior were studied. Naloxone, in doses of 0.3-10 mg/kg (s.c.), antagonized the increase in ambulation and rearing induced by 1 mg/kg of d-amphetamine. When the dose of d-amphetamine was increased to 3 mg/kg, naloxone (3 mg/kg) antagonized only the increase in rearing activity. No dose (0.3-10 mg/kg, s.c.) of naloxone significantly affected d-amphetamine- or apomorphine-induced stereotyped activity. Naloxone (3 mg/kg) significantly augmented the apomorphine (1 mg/kg, s.c.)-induced increase in ambulation but attenuated the apomorphine (0.3 mg/kg)-induced increase in rearing activity. Naloxone (3 mg/kg) or apomorphine (0.03 mg/kg) significantly decreased the ambulation and rearing induced by a novel environment. In combination and in these doses, naloxone and apomorphine produced an additive effect on these behaviors. The neurochemical mechanisms by which naloxone affects d-amphetamine- and apomorphine-induced behavior were investigated. Naloxone (10(-6) M) had no significant effect on [3H]spiroperidol binding in either the caudate nucleus or nucleus accumbens except for a modest inhibition (24%) of both the Km and Bmax in the accumbens microsomal fraction. Similarly, naloxone (10(-6) M) had no significant effect on [3H]dopamine(DA) uptake into either brain region nor did naloxone alter the d-amphetamine-inhibition of uptake. Using perfused tissue slices, naloxone (10(-6)-10(-5) M) significantly attenuated the increase in [3H]DA release induced by d-amphetamine (10(-5) M) in both brain regions. Naloxone (1 mg/kg) had no significant effect on DA or dihydroxyphenyl-acetic acid (DOPAC) levels or on the DA/DOPAC ratio in the caudate nucleus or nucleus accumbens. However, naloxone did reverse the marked increases in the DA-DOPAC ratio induced by d-amphetamine (1 mg/kg) in both brain regions.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1982

The transport and turnover of phospholipids in the rat nigrostriatal system: effects of d-amphetamine and haloperidol.

the nigrostriatal transport of phospholipids was studied using [3H] glycerol, 32Pi and [3H] choline. [3H] glycerol was rapidly incorporated into phospholipid and significant amounts of labelled phospholipid were found in the striatum one hour after injection into the substantia nigra. In contrast, both 32Pi and [3H] choline were more slowly incorporated into phospholipid and significant amounts of labelled phospholipid were not found in the striatum until 24 hours after injection. Once incorporated into the striatum, the [3H-glycerol] phospholipids showed both a rapid (t 1/2 = 1-4 days) and a slow (t 1/2 = 14 + days) turnover component while the [32p] and [3H-Ch] phospholipids showed only a slow turnover component. The subcellular distribution of the rapidly transported [3H-glycerol] phospholipids was studied. Only [3H] phosphatidylcholine (PC) was specifically enriched in the synaptic membrane fraction. The hypothesis was tested that an increase in vesicular lysophosphatidylcholine (LPC) content is associated with dopamine (DA) release. The DA containing vesicles in the striatum were labelled by the intranigral injection of [3H]choline; seven days later, the animals were administered haloperidol to stimulate firing of the nigral-striatal DA neurons. Haloperidol significantly decreased rather than increased the [3H]LPC/[3H]PC ratio. The hypothesis was tested that chronic amphetamine treatment would inhibit phospholipid transport as a result of the decrease in neuronal activity. Chronic d-amphetamine treatment was found to have no effect on the fast component of [3H-glycerol] phospholipid turnover.

Animals↗
PubMed1980

Brain locations controlling the behavioral effects of chronic amphetamine intoxication.

Rats were administered D-amphetamine repeatedly for 4 days. After day 1 of treatment, the amphetamine-induced increases in ambulation, rearing, and stereotyped activity were augmented. However, after 4 days treatment, the rearing and ambulatory responses became attenuated while the stereotyped activities remained augmented. Micro-injection studies revealed that both the augmentation and attenuation of nonstereotyped ambulation were generated from the nucleus accumbens. The augmentation of stereotyped behaviors was generated from the caudate nucleus. Chronically treated animals who were administered 0.7 but not 1.0 mg/kg apomorphine showed augmented behavioral response. Chronic amphetamine treatment significantly decreased (3H) spiroperidol binding in both the nucleus accumbens and caudate nucleus. However, no effect on the DA-stimulated adenyl cyclase activity was observed in either brain region. It is concluded that repeated D-amphetamine administration selectively augments and attenuates D-amphetamine-induced behaviors and that these selective effects are mediated by different dopamine systems.

Adenylyl Cyclases↗
PubMed1980

[Sulfatide: a multifunctional glycolipid constituent of biological membranes].

Sulfogalactosylceramide (or sulfatide) has been localized in the central nervous system by indirect immunofluorescence. This glycosphingolipid belongs essentially to the myelinated areas. Nevertheless, it has also been found in ependymal cells, subpial processes and, in the cerebellum, in the Bergmann fibers. In the brain areas, known to be enriched in opiate receptors, some cells and nerve terminals are also sulfatide positive. In this latter localization, opiates were shown to selectively inhibit binding of the antisulfatide antibodies. We have also shown that antisulfatide inhibited, in vitro, the stereo-specific binding of narcotic drugs and that they antagonized the in vivo effects of morphine and beta-endorphin.

Animals↗