Biomedical subjects

S B Lee

Publications and source records attributed to S B Lee.

At least 73 records · Page 4Linked to original sources

PubMed1999

Comparative study of the inhibition of alpha-glucosidase, alpha-amylase, and cyclomaltodextrin glucanosyltransferase by acarbose, isoacarbose, and acarviosine-glucose.

Bacillus stearothermophilus maltogenic amylase hydrolyzes the first glycosidic linkage of acarbose to give acarviosine-glucose. In the presence of carbohydrate acceptors, acarviosine-glucose is primarily transferred to the C-6 position of the acceptor. When d-glucose is the acceptor, isoacarbose is formed. Acarbose, acarviosine-glucose, and isoacarbose were compared as inhibitors of alpha-glucosidase, alpha-amylase, and cyclomaltodextrin glucanosyltransferase. The three inhibitors were found to be competitive inhibitors for alpha-glucosidase and mixed noncompetitive inhibitors for alpha-amylase and cyclomaltodextrin glucanosyltransferase. The K(i) values were dependent on the type of enzyme and their source. Acarviosine-glucose was a potent inhibitor for baker's yeast alpha-glucosidase, inhibiting 430 times more than acarbose, and was an excellent inhibitor for cyclomaltodextrin glucanosyltransferase, inhibiting 6 times more than acarbose. Isoacarbose was the most effective inhibitor of alpha-amylase and cyclomaltodextrin glucanosyltransferase, inhibiting 15.2 and 2.0 times more than acarbose, respectively.

Acarbose↗

PubMed1999

The Wilms tumor suppressor WT1 encodes a transcriptional activator of amphiregulin.

WT1 encodes a zinc finger transcription factor implicated in kidney differentiation and tumorigenesis. In reporter assays, WT1 represses transcription from GC- and TC-rich promoters, but its physiological targets remain uncertain. We used hybridization to high-density oligonucleotide arrays to search for native genes whose expression is altered following inducible expression of WT1. The major target of WT1 was amphiregulin, a member of the epidermal growth factor family. The WT1(-KTS) isoform binds directly to the amphiregulin promoter, resulting in potent transcriptional activation. The in vivo expression profile of amphiregulin during fetal kidney development mirrors the highly specific pattern of WT1 itself, and recombinant Amphiregulin stimulates epithelial branching in organ cultures of embryonic mouse kidney. These observations suggest a model for WT1 as a transcriptional regulator during kidney differentiation.

Amphiregulin↗

PubMed1999

A novel function of IL-12p40 as a chemotactic molecule for macrophages.

IL-12p70 plays a pivotal role in regulating the Th1/Th2 balance in the initial stage of immune responses. In contrast, IL-12p40, which is produced excess over IL-12p70, has been known to down-regulate IL-12p70-mediated responses by acting as an antagonist. To investigate in vivo function of IL-12p40, RH7777 rat hepatoma cells were engineered to inducibly express mouse IL-12p40 under the tight control of doxycycline (dox). In the absence of dox, s.c. injection of these cells into syngeneic rat was shown to generate tumors. However, the induction of IL-12p40 by dox was sufficient for inhibiting tumor formation, as well as for tumor regression. Immunohistochemical analysis showed that macrophages, but not CD4+ T, CD8+ T, and NK cells, were predominantly recruited into tumor sites as early as 3 days after IL-12p40 induction. These results were further supported by the observation that IL-12p40, but not C-terminal deletion mutants by more than 5 amino acids, was able to chemoattract peritoneal macrophages in vitro, suggesting that IL-12p40, when produced in a large excess over IL-12p70 in vivo, can initially amplify the immune responses against tumors by directly recruiting macrophages. Our findings indicate that IL-12p40 may function as an effector molecule as well as an antagonist of IL-12p70.

Animals↗

PubMed1999

Transient upregulation of osteopontin mRNA in hippocampus and striatum following global forebrain ischemia in rats.

We investigated the spatial and temporal expression of osteopontin (OPN) mRNA following transient forebrain ischemia in rats. Experiments were carried out using a four-vessel occlusion model for forebrain ischemia. The transient induction of OPN mRNA after global ischemia occurred earlier in the striatum than in the hippocampus. It was pronounced in the dorsomedial striatum close to the lateral ventricle and in the CA1 subfield and the subiculum of the hippocampus before microglial cells became more reactive. It also could be detected in the dentate hilus and to a marginal extent in the CA3. Our results suggest that the hippocampus and the striatum following global forebrain ischemia upregulate OPN mRNA in different spatiotemporal profiles.

Animals↗

PubMed1999

Inhibitory effect of mineral ion accumulation on high density growth of the hyperthermophilic archaeon Sulfolobus solfataricus.

A fed-batch operation for high density cultivation of Sulfolobus solfataricus (DSM 1617) in a bench-top fermentor using a feed medium composed of glucose and yeast extract was investigated. The highest maximal cell density obtained in controlled fed-batch cultures was 21.7 g/l. Although higher yeast extract concentrations in the medium favored greater cell biomass yield, cell growth ceased with low cell densities. It was observed that large amounts of inorganic ions, such as sulfate, ammonium, potassium and phosphate ions, were accumulated in the culture broth at higher yeast extract concentrations. This was due to either the addition of the titrant or feeding of yeast extract during cultivation. Fed-batch cultures with additional mineral salts in the feed medium showed much lower cell biomass, indicating that accumulation of inorganic ions has a significant inhibitory effect on the growth of S. solfataricus. Inhibition of cell growth by the presence of mineral ions was further confirmed by the batch culture experiments. Some plausible mechanisms which can account for the growth inhibition at higher mineral ion concentrations have been suggested.

Journal Article↗

PubMed1999

Differential expression and regulation of TGF-beta1, TGF-beta2, TGF-beta3, TGF-betaRI, TGF-betaRII and TGF-betaRIII in cultured human corneal, limbal, and conjunctival fibroblasts.

PURPOSE: We have reported that three patterns of cytokine expression are potentially involved between epithelia and fibroblasts of the human ocular surface. The TGF-beta family is a prototypical fibrogenic cytokine responsible for fibroblast activation in wound healing. We investigated how the TGF-beta family is differentially expressed and regulated in cultured human corneal, limbal and conjunctival fibroblasts. METHODS: Human corneal (HCF), limbal (HLF) and conjunctival fibroblast (HJF) were cultured in DMEM-10% FBS until confluence and switched to serum-free DMEM-ITS for 48 h before adding 10 ng/ml of each of eight cytokines for 4 h in three separate experiments. Total RNA was isolated and subjected to Northern hybridization with GAPDH as a control. ELISA was used to determine TGF-beta1 and TGF-beta2 proteins in the media. RESULTS: All three isoforms of TGF-beta and three types of TGF-betaR were expressed by HCF, HLF and HJF. Expression of TGF-beta1 mRNA was strongest and upregulated by the three TGF-betas in all three types of fibroblast. PDGF-BB and TGF-alpha slightly increased TGF-beta1 mRNA. TGF-betas also upregulated TGF-beta3 mRNA in HJF. TGF-betaRI mRNA was the only receptor upregulated by TGF-betas. TGF-betaRII and TGF-betaRIII mRNA were not regulated by all cytokines tested. CONCLUSIONS: TGF-betas auto-induction is the major mechanism upregulating TGF-beta1 expression. Promotion of TGF-beta3 by the TGF-betas may have a special role in HJF. Differential expression and regulation of TGF-betas and TGF-betaRs suggest that each TGF-beta isoform may have specific functions in different ocular surface fibroblasts. No cytokine tested can downregulate TGF-beta1 and the TGF-betaRs.

Adult↗

PubMed1999

Comparison of inferior oblique muscle weakening by anterior transposition or myectomy: a prospective study of 20 cases.

BACKGROUND/AIMS: Among the various weakening techniques of inferior oblique muscle overaction, the most commonly used techniques include myectomy, recession, and anterior transposition. Anterior transposition and myectomy were compared to evaluate the surgical results in inferior oblique overaction. METHODS: 20 children with bilateral +3 overacting inferior oblique muscles underwent a prospective randomised study by which the anterior transposition procedure in one eye was compared with the myectomy procedure in the other eye. RESULTS: Postoperative follow up averaged 2 years. The success rates in two surgical procedures were 85% for the anterior transposition and 25% for the myectomy (standard of success was based on zero inferior oblique overaction). In only one case did the anterior transposition tend to limit the elevation of the eye in the midline, adduction, and abduction. Anterior transposition produced hypotropia at the primary position in only one case. Most eyes that underwent myectomy (75%) showed apparent residual overaction. CONCLUSION: The anterior transposition appeared to be more effective in eliminating the overaction of inferior oblique muscle than the myectomy.

Child↗

PubMed1999

Incidence of eyelid cancers in Singapore from 1968 to 1995.

AIM: To describe the epidemiological characteristics of patients with eyelid malignancies seen in all hospitals in Singapore from 1968 to 1995. METHOD: The Singapore Cancer Registry has been collecting epidemiological data of all cancers seen in Singapore since 1968. The data of all cases of Singapore residents with eyelid cancers diagnosed from 1968 to 1995 (ICD-9, sites 172.1 and 173.1) were retrieved for analysis. RESULTS: There were 162 male patients (49.8%) and 163 females (50.2%). The median age at diagnosis was 63 years in males and 66 years in females. The average annual age standardised incidence rate among male Singapore residents was 6.5 per million and 5.5 per million among female Singapore residents. Between 1993 and 1995, the average annual rate for females was 6.8 per million, compared with 3.1 per million between 1968 and 1972. The most common cancer was basal cell carcinoma (84.0%), followed by sebaceous adenocarcinoma (10.2%) and squamous cell carcinoma (3.4%). CONCLUSION: The annual age standardised incidence for male residents has remained relatively stable. The incidence for female residents has shown a steady increase over the past 28 years. The incidence for males is generally higher than that for females. These expanded epidemiological characteristics may serve to provide a foundation to monitor future disease patterns and to promote further research into the aetiology of these cancers.

Adolescent↗

PubMed1999

Biochemical and pharmacological characteristics of a newly synthesized H+/K+-ATPase inhibitor, YJA20379-8, 3-butyryl-4-[R-1-methylbenzylamino]-8-ethoxy-1,7-naphthyridine, in pigs and rats.

We have investigated the effect of the newly synthesized proton-pump inhibitor YJA20379-8, 3-butyryl-4-[R-1-methylbenzylamino]-8-ethoxy-1,7-naphthyridine, on gastric mucosal proton pump (H+/K+-ATPase) activity, gastric acid secretion and gastric lesions in experimental animals. In lyophilized pig gastric microsomes, YJA20379-8 was shown to inhibit H+/K+-ATPase activity; the inhibitory effect was not affected by pH, the IC50 (dose resulting in 50% inhibition) being 28.0 microM and 30.0 microM at pH 6.4 and pH 7.4, respectively. The effect was fully reversed by dilution and subsequent washing of the incubation mixtures of H+/K+-ATPase and YJA20379-8, suggesting the reversible nature of the enzyme inhibition. In pylorus-ligated rats, YJA20379-8 administered by different routes (intraduodenal, subcutaneous, intravenous or oral) resulted in dose-dependent suppression of basal gastric acid secretion. The duration of antisecretory action of 30 mg kg(-1) YJA20379-8 given intraduodenally was very brief (less than 7 h). Pretreatment with YJA20379-8 also dose-dependently prevented gastric lesions induced by absolute ethanol and water-immersion stress in rats. These results suggest that YJA20379-8 might exert its antiulcer activity partly by reversible suppression of acid secretion and partly by protecting the gastric mucosa against ulcerative stimuli.

Animals↗

PubMed1999

Management of subluxated crystalline lenses with planned intracapsular cataract extraction and anterior chamber intraocular lens implantation.

PURPOSE: To review the visual and surgical outcome of planned intracapsular cataract extraction (ICCE) and anterior chamber intraocular lens (AC IOL) implantation for subluxated crystalline lenses. METHODS: Eight eyes of 7 patients with subluxated lenses which underwent planned ICCE, AC IOL implantation and peripheral iridectomy by one surgeon between September 1995 and November 1996 were prospectively followed-up to ascertain their visual and surgical outcome. RESULTS: There were 4 male and 3 female patients. Their mean age was 64.4 years (range, 39-86 years). Seven lenses were removed by cryoextraction while one was removed with the aid of a vectis. The mean post-operative follow-up was 11.6 months, (range, 3-22 months). Excluding 1 eye with pre-existing ocular pathology, all eyes achieved a best corrected visual acuity (BCVA) of 6/12 or better post-operatively. One of these eyes, in a schizophrenia patient, developed subluxation of the AC IOL and retinal detachment 11 months after the cataract surgery due to repeated eye rubbing. The final BCVA became 6/120 after successful repositioning of the AC IOL and retinal re-attachment surgery. One eye achieved a BCVA of 6/36 due to age-related macular degeneration. CONCLUSION: Our results show that planned ICCE and AC IOL implantation is a useful and safe procedure in the management of subluxated lenses.

Adult↗

PubMed1999

Biochemical and pharmacological characteristics of a newly synthesized H+-K+ ATPase inhibitor, YJA20379-1, 2-amino-4,5-dihydro-8-phenylimidazole[2,1-b]thiazolo[5,4-g]benzothiazol e.

The biochemical and pharmacological characteristics of a newly synthesized H+-K+ ATPase inhibitor, 2-amino-4,5-dihydro-8-phenylimidazole[2,1-b]thiazolo[5,4-g]benzothiazole (YJA20379-1), were investigated. In the pig gastric microsomes, YJA20379-1 inhibited the gastric H+-K+ ATPase regardless of pH condition, IC50 values being 21 and 24 microM at pH 6.4 and 7.4, respectively. The inhibitory activity of YJA20379-1 was antagonized by dithiothreitol treatment but could not be reversed by dilution and washing of the enzyme preparation. In Sprague-Dawley rats, YJA20379-1, administered i.d., p.o, i.v., or s.c., significantly inhibited basal gastric acid secretion, with ED50 values of 4.7, 20.2, 6.3, and 13.4 mg/kg, respectively. The antisecretory action of YJA20379-1 was short lasting (less than 7 h at an oral dosing of 30 mg/kg). Oral administration of YJA20379-1 also prevented the formation of ethanol, indomethacin, and water immersion stress induced gastric lesions and mepirizole-induced duodenal ulcers in rats. Furthermore, YJA20379-1 accelerated the healing of acetic acid induced chronic gastric ulcers in rats. In conclusion, these results suggest that YJA20379-1 has a potent inhibitory activity on the gastric H+-K+ ATPase but much shorter duration of antisecretory action than omeprazole, thereby exerting its anti-ulcer effects partly with cytoprotective activity.

Animals↗

PubMed1998

Targeted disruption of the Ca2+ channel beta3 subunit reduces N- and L-type Ca2+ channel activity and alters the voltage-dependent activation of P/Q-type Ca2+ channels in neurons.

In comparison to the well characterized role of the principal subunit of voltage-gated Ca2+ channels, the pore-forming, antagonist-binding alpha1 subunit, considerably less is understood about how beta subunits contribute to neuronal Ca2+ channel function. We studied the role of the Ca2+ channel beta3 subunit, the major Ca2+ channel beta subunit in neurons, by using a gene-targeting strategy. The beta3 deficient (beta3-/-) animals were indistinguishable from the wild type (wt) with no gross morphological or histological differences. However, in sympathetic beta3-/- neurons, the L- and N-type current was significantly reduced relative to wt. Voltage-dependent activation of P/Q-type Ca2+ channels was described by two Boltzmann components with different voltage dependence, analogous to the "reluctant" and "willing" states reported for N-type channels. The absence of the beta3 subunit was associated with a hyperpolarizing shift of the "reluctant" component of activation. Norepinephrine inhibited wt and beta3-/- neurons similarly but the voltage sensitive component was greater for N-type than P/Q-type Ca2+ channels. The reduction in the expression of N-type Ca2+ channels in the beta3-/- mice may be expected to impair Ca2+ entry and therefore synaptic transmission in these animals. This effect may be reversed, at least in part, by the increase in the proportion of P/Q channels activated at less depolarized voltage levels.

Animals↗

PubMed1998

Analysis of the gene encoding cyclomaltodextrinase from alkalophilic Bacillus sp. I-5 and characterization of enzymatic properties.

The gene encoding cyclomaltodextrinase (CDase) was cloned from alkalophilic Bacillus sp. I-5. The nucleotide sequence of the gene was determined and the physicochemical properties of the enzyme were investigated. The gene had an open reading frame of 559 amino acids with a predicted molecular weight of 64,884. The enzyme was purified to near homogeneity from Escherichia coli cells carrying a recombinant plasmid that contained the CDase gene. The enzyme hydrolyzed cyclomaltoheptaose (beta-CD) 13 times better than starch and 33 times better than pullulan, and it had transglycosylation activity. The enzyme also hydrolyzed acarbose, a pseudotetrasaccharide inhibitor of glucosidases. The enzyme was stabilized by Ca2+ and the activity was increased more than twofold in the presence of 5 mM EDTA. The optimum temperature of the enzyme was elevated from 40 to 50 degrees C by Ca2+ ion and the thermal activity was maintained more than 80% at 60 degrees C in the presence of Ca2+. Comparison of known amino acid sequences of several amylolytic enzymes with cyclomaltodextrinase activity, site-directed mutagenesis of the enzyme, and substrate specificity of the enzyme imply that the region between the third and the fourth conserved regions of the enzyme may play an important role in binding and degradation of cyclomaltodextrin.

Acarbose↗

PubMed1998

E1B 55K sequesters WT1 along with p53 within a cytoplasmic body in adenovirus-transformed kidney cells.

WT1 encodes a tumor suppressor that is expressed in cells of the developing kidney and is inactivated in Wilms tumor, a pediatric kidney cancer. The adenovirus E1B 55K gene product contributes to the transformation of primary baby rat kidney (BRK) cells by binding and inactivating the product of the p53 tumor suppressor. We have previously demonstrated that WT1 and p53 are present within a protein complex in vivo. We now show that WT1 is physically associated with E1B 55K in adenovirus-transformed cells, an interaction that is mediated by the first two zinc fingers of WT1. Immunodepletion of p53 abrogates the coimmunoprecipitation of E1B 55K and WT1, consistent with the presence of a trimeric protein complex containing these three proteins. In the presence of E1B 55K, WT1 which is normally localized in the nucleus, is retained within a very high molecular weight complex and sequestered in the characteristic perinuclear cytoplasmic body that contains E1B 55K and p53. Expression of E1B 55K in osteosarcoma cells that undergo apoptosis following expression of WT1 inhibits WT1-mediated cell death. We conclude that E1B 55K may target WT1 along with p53, resulting in the functional inactivation of both tumor suppressor gene products by this viral oncoprotein.

Adenoviridae↗

PubMed1998

Inhibition of cellular proliferation by the Wilms tumor suppressor WT1 requires association with the inducible chaperone Hsp70.

The Wilms tumor suppressor WT1 encodes a zinc finger transcription factor that is expressed in glomerular podocytes during a narrow window in kidney development. By immunoprecipitation and protein microsequencing analysis, we have identified a major cellular protein associated with endogenous WT1 to be the inducible chaperone Hsp70. WT1 and Hsp70 are physically associated in embryonic rat kidney cells, in primary Wilms tumor specimens and in cultured cells with inducible expression of WT1. Colocalization of WT1 and Hsp70 is evident within podocytes of the developing kidney, and Hsp70 is recruited to the characteristic subnuclear clusters that contain WT1. The amino-terminal transactivation domain of WT1 is required for binding to Hsp70, and expression of that domain itself is sufficient to induce expression of Hsp70 through the heat shock element (HSE). Substitution of a heterologous Hsp70-binding domain derived from human DNAJ is sufficient to restore the functional properties of a WT1 protein with an amino-terminal deletion, an effect that is abrogated by a point mutation in DNAJ that reduces binding to Hsp70. These observations indicate that Hsp70 is an important cofactor for the function of WT1, and suggest a potential role for this chaperone during kidney differentiation.

Amino Acid Sequence↗

PubMed1998

Isolation of a new member of DnaJ-like heat shock protein 40 (Hsp40) from human liver.

A new member of Hsp40, HLJ1, consisting of 337 amino acids, was cloned from a human liver cDNA library. The deduced amino acid sequence of HLJ1 has an 84% homology (69% identity) with that of HDJ-1 isolated from human placenta. Northern analysis showed that expression of the HLJ1 gene is heat-inducible and its transcription shows some degree of preference in heart, skeletal muscle, and pancreas.

Amino Acid Sequence↗

PubMed1998

Dopamine transporter density measured by [123I]beta-CIT single-photon emission computed tomography is normal in dopa-responsive dystonia.

The clinical distinction between dopa-responsive dystonia (DRD) and juvenile Parkinson's disease JPD) can pose a diagnostic challenge. Both conditions are dopa responsive. However, long-term L-dopa benefit is very different between the two. The difference in the prognosis is due to presence or absence of nigral cell loss. In JPD, there is degenerative nigral cell loss, whereas there are enzymatic defects in dopamine synthesis without cell loss in DRD. Mutations have been found in the GTP cyclohydrolase I (GCH-I) and tyrosine hydroxylase genes in DRD. As the discovered mutations are multiple and more are expected to be found, it is difficult to confirm or exclude DRD by mutation studies. Measurement of cerebrospinal fluid (CSF) neopterin will detect DRD from mutations in the GCH-I gene but not from mutations in tyrosine hydroxylase. The dopamine transporter (DAT) is a protein in the dopaminergic nerve terminals. (1R)-2beta-Carbomethoxy-3beta-(4-[123I]iodophenyl)tropane ([123I]beta-CIT) is a ligand for the DAT, and it was shown to be a useful nuclear imaging marker for neurons that degenerate in Parkinson's disease (PD). As DRD was shown to have a normal DAT without nigral cell loss in a postmortem study, we predicted that the DAT measured in vivo by nuclear imaging will be normal in DRD and will differentiate DRD from JPD. Therefore, we performed [123I]beta-CIT single-photon emission computed tomography ([123I]beta-CIT SPECT) in clinically diagnosed DRD, PD, and JPD, and examined whether DAT imaging can differentiate DRD from PD and JPD. We then examined whether DAT imaging can provide a screening tool for molecular genetic studies, by studying mutations in the candidate gene GCH-I and measuring CSF neopterin. Five females (4 from two families, and 1 sporadic) were diagnosed as DRD based on early-onset foot dystonia and progressive parkinsonism beginning at ages 7 to 12. All patients were functioning normally on L-dopa 100 to 250 mg/day for up to 8 years. SPECT imaging was obtained after intravenous injection of [123I]beta-CIT; 15 healthy volunteers served as normal control, and 6 PD and 1 JPD as disease controls. [123I]beta-CIT striatal binding was normal in DRD, whereas it was markedly decreased in PD and JPD. Gene analysis showed a novel nonsense mutation in the GCH-I gene in one family. No mutation was found in the other family or in the sporadic case. CSF neopterin was markedly decreased in the 4 tested patients. [123I]beta-CIT SPECT is a sensitive method for probing the integrity of nigrostriatal dopaminergic nerve terminals. A normal striatal DAT in a parkinsonian patient is evidence for a nondegenerative cause of parkinsonism and differentiates DRD from JPD. Finding a new mutation in one family and failure to demonstrate mutations in the putative gene in other cases supports the usefulness of DAT imaging in diagnosing DRD.

Adult↗

PubMed1998

Expression of Bfl-1 in normal and tumor tissues: Bfl-1 overexpression in cancer is attributable to its preferential expression in infiltrating inflammatory cells.

Bfl-1, a member of the Bcl-2 gene family, blocks p53-mediated apoptosis and has oncogenic transforming activity. In normal tissues, the transcript of Bfl-1 is expressed abundantly in bone marrow and at a low level in several other tissues. In previous experiments, elevated expression of Bfl-1 was observed by Northern analysis of stomach cancer samples. To study the role of Bfl-1 in normal cell development and in tumorigenesis, we have analyzed the expression of Bfl-1 in normal and tumor tissues by the in situ hybridization technique. The Bfl-1 transcript was detected in the white pulp of the spleen and in the germinal center of lymphatic tissues. In tumor tissues, its expression was preferentially detected in infiltrating inflammatory cells rather than in cancer cells, suggesting that Bfl-1 is not involved in tumorigenesis.

Colonic Neoplasms↗