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Biomedical subjects

B Kim

Publications and source records attributed to B Kim.

At least 73 records · Page 4Linked to original sources

Detection of the cholera toxin-binding activity of kappa-casein macropeptide and optimization of its production by the response surface methodology.

The cholera toxin (CT)-binding activity of purified kappa-casein macropeptide (CMP) from bovine kappa-casein was detected. In addition, a statistical model was developed to optimize the production of CMP. CMP was prepared by chymosin hydrolysis of kappa-casein and a subsequent 3% trichloroacetic acid treatment. CMP was further fractionated in an ion-exchange column by FPLC. CT binding activity was eluted at 0.18 M NaCl and was a single 8.9 kDa peptide without tyrosine and arginine residues. The CT binding activity was rapidly lost by a carbohydrase treatment. The conditions for CMP production with chymosin were optimized by using the response surface methodology (RSM). The estimated optimum levels of the factors were as follows: reaction temperature, 38.5 degrees C; pH, 6.44; and time, 35.9 min. A validation experiment was performed in which CMP was prepared under the predicted parameters, and it was ascertained that the estimated optimum conditions gave better production of CMP than any other conditions.

Animals↗

Molecular epidemiology of astroviruses in Japan from 1995 to 1998 by reverse transcription-polymerase chain reaction with serotype-specific primers (1 to 8).

In addition to the serotype-specific primers described previously (1 to 7), a new serotype 8-specific primer has been designed, allowing detection of all astrovirus serotypes. A total of 1,382 diarrheal stool samples in 5 regions in Japan were examined by reverse transcription-polymerase chain reaction (RT-PCR). The incidence of astrovirus infection in all 5 regions was 5.9% (82 of 1,382 samples) and infection occurred mainly from November to April. Serotypes 1, 3, and 4 were detected in 66, 14, and 2 of the 82 positive samples, respectively. None of the other serotypes was detected. The highest detection rate was from 0 to 1 year old, 39.0%, and the next highest was from 1 to 2 years old, 34.1%. The primers provide a useful approach for study of the epidemiology of astroviruses.

Astroviridae Infections↗

Isolation and characterization of a thermophilic lipase from bacillus thermoleovorans ID-1.

A thermophilic microorganism, Bacillus thermoleovorans ID-1, isolated from hot springs in Indonesia, showed extracellular lipase activity and high growth rates on lipid substrates at elevated temperatures. On olive oil (1.5%, w/v) as the sole carbon source, the isolate ID-1 grew very rapidly at 65 degrees C with its specific growth rate (2.50 h(-1)) and its lipase activity reached the maximum value of 520 U l(-1) during the late exponential phase and then decreased. In addition to this, isolate ID-1 could grow on a variety of lipid substrates such as oils (olive oil, soybean oil and mineral oil), triglycerides (triolein, tributyrin) and emulsifiers (Tween 20, 40). The excreted lipase of ID-1 was purified 223-fold to homogeneity by ammonium sulfate precipitation, DEAE-Sephacel ion-exchange chromatography and Sephacryl S-200 gel filtration chromatography. As a result, the relative molecular mass of the lipase was determined to be 34 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme showed optimal activity at 70-75 degrees C and pH 7.5 and exhibited 50% of its original activity after 1 h incubation at 60 degrees C and 30 min at 70 degrees C and its catalytic function was activated in the presence of Ca(2+) or Zn(2+).

Bacillus↗

Characterization of subtype A HIV-1 from Africa by full genome sequencing.

OBJECTIVE: To improve our understanding of the genetic complexity of HIV-1 subtype A by increasing the number of subtype A isolates that have been sequenced in their entirety. METHODS: Nine HIV-1-seropositive patients from Africa living in Sweden contributed peripheral blood mononuclear cells (PBMC) for this study. Sequencing of the C2-V3 region of env had shown them to be subtype A. DNA from virus cultures was used for the amplification of virtually full-length proviral sequences, and the resulting fragment was sequenced. RESULTS: Six of the nine viral isolates were subtype A throughout the genome, or non-recombinant, and all of these were from east Africa. One virus from the Ivory Coast had the AG(IbNG) genetic form, a recombinant form common in west Africa. Two of the isolates were novel recombinants: one was an A/C recombinant and the other was A/D. Analysis of gag reveals three subclusters within the A subtype: one containing the AG(IbNG) subtype viruses, one containing the AE(CM240) viruses and one containing the non-recombinant A viruses. These genetic clusters have different geographical distributions in Africa. CONCLUSION: The prevailing view of HIV-1 subtype A forming a uniform band across the center of sub-Saharan Africa needs revision. In all probability, the most common subtype in west Africa and west central Africa is the AG recombinant, AG(IbNG), whereas in east central Africa it is the non-recombinant subtype A.

Africa↗

New human immunodeficiency virus, type 1 reverse transcriptase (HIV-1 RT) mutants with increased fidelity of DNA synthesis. Accuracy, template binding, and processivity.

Infidelity of DNA synthesis by human immunodeficiency virus, type 1 reverse transcriptase (HIV-1 RT) is a presumptive determinant of HIV-1 hypervariability and is incompletely understood at the mechanistic and structural levels. Amino acid substitution at only three residues, including Asp-76 (Kim, B., Hathaway, T. R., and Loeb, L. A. (1996) Biochemistry 37, 5831-5839), is known to increase fidelity. We report here that substitution at Arg-78 can also increase accuracy. Mutant R78A RT showed reduced primer extension in misincorporation assays lacking a complementary dNTP and exhibited a 9-fold decrease in mutation frequency in the M13mp2 lacZ forward mutation assay. Previous structural studies indicate that Arg-78 and Asp-76 lie in a region that interacts with template nucleotides. Interestingly, R78A RT exhibited 6- to 8-fold decreases in binding affinity (K(d)) for RNA and DNA templates relative to wild type RT. In contrast, D76V RT, which also increases fidelity (Kim et al., 1996), showed a 6- to 7-fold increased affinity. The processivity of R78A RT on both RNA and DNA templates was substantially reduced relative to wild type RT, whereas the processivity of D76V RT was increased. We discuss relationships of fidelity, template binding, and processivity in these and other HIV RT mutants.

Amino Acid Substitution↗

Maturational changes in cerebral lactate and acid clearance following ischemia measured in vivo using magnetic resonance spectroscopy and microdialysis.

Intraischemic hyperglycemia has different effects on neurologic outcome in mature vs. immature brain, and may reflect differences in the extent or duration of cerebral lactic acidosis. We examined the hypotheses that post-ischemic lactate and acid clearance rates depend on the severity of intraischemic cerebral acidosis, and that rates of clearance change as a function of brain maturation. In vivo 31P and 1H magnetic resonance spectroscopy (MRS) was used to compare intracellular acid and lactate clearance rates in newborn and 1-month old swine following a 14-min episode of transient near-complete global ischemia. In the same animals, in vivo microdialysis was used to determine if extracellular lactate clearance changed as a function of cerebral lactic acidosis or differed between age groups following ischemia. Plasma glucose concentration was altered in individual animals to study a range of intraischemic cerebral lactic acidosis. For both age-groups, maximal brain acidosis and lactosis occurred in the post-ischemia interval, indicating a delay in the re-establishment of oxidative metabolism following ischemia. Clearance half-lives of both cerebral acidosis and lactosis increase as a function of increased intraischemic cerebral acidosis. For either age group, the clearance half-life for acidosis was faster than the half-life for lactate. However, the subgroup of 1-month old swine who experienced severe cerebral acidosis (i.e., pH<6.1) had a longer cerebral lactate clearance half-life as compared to the subgroup of newborn animals with a similar severity of acidosis. In both age groups, there were comparable maximal increases in extracellular lactate concentrations in the post-ischemic period and similar rates of decline from the maximum. These results demonstrate that post-ischemic lactate and acid clearance are altered by the extent of intraischemic acidosis, and the extent of post-ischemic uncoupling between brain acid and lactate clearance increases with advancing age. The transmembrane clearance of lactate was not a prominent mechanism that differentiated lactate clearance rates between newborn and 1-month old swine.

Acidosis, Lactic↗

Motion correction in fMRI via registration of individual slices into an anatomical volume.

An automated retrospective image registration based on mutual information is adapted to a multislice functional magnetic resonance imaging (fMRI) acquisition protocol to provide accurate motion correction. Motion correction is performed by mapping each slice to an anatomic volume data set acquired in the same fMRI session to accommodate inter-slice head motion. Accuracy of the registration parameters was assessed by registration of simulated MR data of the known truth. The widely used rigid body volume registration approach based on stacked slices from the time series data may hinder statistical accuracy by introducing inaccurate assumptions of no motion between slices for multislice fMRI data. Improved sensitivity and specificity of the fMRI signal from mapping-each-slice-to-volume method is demonstrated in comparison with a stacked-slice correction method by examining functional data from two normal volunteers. The data presented in a standard anatomical coordinate system suggest the reliability of the mapping-each-slice-to-volume method to detect the activation signals consistent between the two subjects.

Algorithms↗

Phylogenetic relationships among Hepatozoon species from snakes, frogs and mosquitoes of Ontario, Canada, determined by ITS-1 nucleotide sequences and life-cycle, morphological and developmental characteristics.

The molecular biological characteristics of Hepatozoon species infecting various species of snakes, frogs and mosquitoes were investigated by determining the nucleotide sequences of the first internal transcribed spacer region. A phylogenetic analysis was performed on seven isolates of Hepatozoon infecting snakes, including Hepatozoon sipedon and four morphologically similar but not identical forms, and two isolates of Hepatozoon catesbianae infecting Green frogs (Rana clamitans melanota). This analysis, which utilised data from first internal transcribed spacer nucleotide sequences, morphological and morphometric features of gamonts, oocysts and sporocysts, and previously determined life-cycle and host-specificity characteristics, revealed that H. sipedon is a polymorphic species with a wide host and geographic range. Four synapomorphies. including two nucleotide substitutions and two morphological character state changes, supported a monophyletic group of six isolates of H. sipedon from the central region of Ontario which was the sister group for an isolate (HW1) from the southern part of the province. Based on the results of this study, an evaluation of which criteria are useful for describing species of Hepatozoon is presented, with the intent of curtailing the practice of naming species based on morphological features of gamonts or on incomplete life-cycle data.

Animals↗

Semiautomatic registration of volumetric ultrasound scans.

We demonstrate the ability to register easily and accurately volumetric ultrasound scans without significant data preprocessing or user intervention. Two volumetric ultrasound breast scan data sets were acquired from two different patients with breast cancer. Volumetric scan data were acquired by manually sweeping a linear array transducer mounted on a linear slider with a position encoder. The volumetric data set pairs consisted of color flow and/or power mode Doppler data sets acquired serially on the same patients. A previously described semiautomatic registration method based on maximizing mutual information was used to determine the transform between data sets. The results suggest that, even for the deformable breast, three-dimensional full affine transforms can be sufficient to obtain clinically useful registrations; warping may be necessary for increased registration accuracy. In conclusion, mutual information-based automatic registration as implemented on modern workstations is capable of yielding clinically useful registrations in times <35 min.

Breast Neoplasms↗

Classification of thermophilic streptomycetes, including the description of Streptomyces thermoalcalitolerans sp. nov.

A polyphasic taxonomic study was undertaken to clarify relationships within and between representative thermophilic alkalitolerant streptomycetes isolated from soil and appropriate marker strains. The resultant data, notably those from DNA-DNA relatedness studies, support the taxonomic integrity of the validly described species Streptomyces thermodiastaticus, Streptomyces thermoviolaceus and Streptomyces thermovulgaris. However, the genotypic and phenotypic data clearly show that Streptomyces thermonitrificans Desai and Dhala 1967 and S. thermovulgaris (Henssen 1957) Goodfellow et al. 1987 represent a single species. On the basis of priority, S. thermonitrificans is a later subjective synonym of S. thermovulgaris. Similarly, 10 out of the 11 representative thermophilic alkalitolerant isolates had a combination of properties consistent with their classification as S. thermovulgaris. The remaining thermophilic alkalitolerant isolate, Streptomyces strain TA56, merited species status. The name Streptomyces thermoalcalitolerans sp. nov. is proposed for this strain. A neutrophilic thermophilic isolate, Streptomyces strain NAR85, was identified as S. thermodiastaticus.

DNA, Bacterial↗

Streptomyces malaysiensis sp. nov., a new streptomycete species with rugose, ornamented spores.

The taxonomic position of a streptomycete strain isolated from Malaysian soil was established using a polyphasic approach. The organism, designated strain ATB-11T, was found to have chemical and morphological properties consistent with its classification in the genus Streptomyces. An almost complete 16S rRNA gene (rDNA) sequence determined for the test strain was compared with those of previously studied streptomycetes by using two treeing algorithms. The 16S rDNA sequence data not only supported classification of the strain in the genus Streptomyces but also showed that it formed a distinct phyletic line. At maturity, the aerial hyphae of strain ATB-11T differentiated into tight, spiral chains of rugose, cylindrical spores. The organism was readily distinguished from representatives of validly described Streptomyces species with rugose spores by using a combination of phenotypic features. It is proposed, therefore, that strain ATB-11T be classified in the genus Streptomyces as Streptomyces malaysiensis sp. nov.

Bacterial Typing Techniques↗

Orientational relaxation in Brownian rotors with frustrated interactions on a square lattice.

We present simulation results on the equilibrium relaxation of Brownian planar rotors based on a uniformly frustrated XY model on a square lattice. The rotational relaxation exhibits typical dynamic features of fragile supercooled liquids including the two-step relaxation. We observe a dynamic crossover from the high-temperature regime with Arrhenius behavior to the low-temperature regime with temperature-dependent activation energy. A consistent picture for the observed slow dynamics can be given in terms of the caging effect and thermal activation across potential barriers in the energy landscapes.

Journal Article↗

Endoscopic and histological patchiness in treated ulcerative colitis.

OBJECTIVE: Traditionally, contiguous distribution of inflammation (endoscopic and histological) with rectal involvement is thought to be important in distinguishing ulcerative colitis (UC) from Crohn's disease of the colon. Little long-term data are available that prove whether this rule holds during the course of disease as it is modified by time and treatment. The aim of this study was to investigate the prevalence of endoscopic and histological patchiness and rectal sparing in treated UC over time and to correlate these findings with treatment at the time of endoscopy. METHODS: Patients with well-established UC who underwent sequential colonoscopy or flexible sigmoidoscopy with biopsies were included in this study. Patients' medical records including endoscopy/biopsy reports and clinical status/symptoms/treatment at the time of endoscopy were reviewed retrospectively. RESULTS: A total of 32 patients (14 men, 18 women; median age, 45 yr; median UC duration, 15 yr) underwent 175 sequential endoscopies with biopsies (161 colonoscopies, 14 sigmoidoscopies; median, five endoscopies per patient; range, 3-10). Endoscopic and/or histological patchiness was present in 20 of 175 (11%) sequential endoscopies with biopsies over time from 12 of 32 (38%) patients. Endoscopic and/or histological rectal sparing was present in 27 of 175 (15%) of sequential endoscopies with biopsies over time from 14 of 32 (44%) patients. Seven patients had both patchiness and rectal sparing. Therefore, in 47 (27%) follow-up endoscopies in 19 (59%) patients, there was either patchy disease, rectal sparing, or both sometime during the course of disease with treatment. There was no significant difference in treatment, including steroid use and rectal therapy, between those with patchiness and/or rectal sparing and those without. CONCLUSIONS: Contrary to traditional teaching, endoscopic and histological patchiness of inflammation and rectal sparing are common during the course of disease in treated UC and seem to be unrelated to specific therapy.

Adolescent↗

Serotypes of human rotaviruses in 7 regions of Japan from 1984 to 1997.

Human rotavirus (HRV) serotypes were studied from diarrheal stool specimens in children in 7 regions of Japan (Sapporo, Tokyo, Maizuru, Osaka, Kagawa, Kurume, and Saga) from 1984 to 1997 by enzyme immunoassay (EIA) with serotype-specific monoclonal antibodies against serotypes 1, 2, 3, and 4. In addition, reverse transcription-polymerase chain reaction (RT-PCR) was conducted for analysis of "others" which included nonserotypable and mixed-serotype rotavirus specimens by EIA. In 3756 rotavirus-positive specimens, serotype 1 was detected in 2649 (70.5%), serotype 2 in 362 (9.6%), serotype 3 in 232 (6.2%) and serotype 4 in 196 (5.2%). Overall, serotype 1 was predominant from 1984 to 1997, although there were a few cases in which serotype 2, 3 and 4 became predominant based on area and year. The frequency of serotype 1 has gradually increased since 1993. Twenty two, 2, 3 and 1 among 57 specimens of "others" by EIA from Tokyo, Maizuru, Sapporo and Kurume in 1995-1996 and 1996-1997 were determined as serotypes 1, 2, 3, and 9 by RT-PCR, respectively.

Child↗

In situ hybridization for the detection and localization of swine Chlamydia trachomatis.

Gnotobiotic piglets were inoculated intralaryngeally with swine Chlamydia trachomatis strain R33 or orally with swine C. trachmatis strain R27. Archived formalin-fixed, paraffin-embedded tissues from piglets euthanatized 4-7 days postinoculation were examined by in situ hybridization for C. trachomatis nucleic acid using a nonradioactive digoxigenin-labeled DNA probes that targeted specific ribosomal RNA or omp1 mRNA molecules of the swine C. trachomatis strains. Positive hybridization signals were detected in bronchial epithelial cells, bronchiolar epithelial cells, pneumocytes, alveolar and interstitial macrophages, and jejunal and ileal enterocytes. Chlamydia-infected cells had a strong signal that was confined to the intracytoplasmic inclusions. Positive hybridization signals were not detected in tissue sections from an uninfected control piglet or in C. psittaci-infected sheep placenta. The morphology of host cells was preserved despite the relatively high temperature required in parts of the incubation procedure. The data indicate that in situ hybridization can be used to detect swine C. trachomatis in formalin-fixed, paraffin-embedded tissue specimens.

Animals↗

Primary mucinous cystadenocarcinoma of the appendix: CT findings.

OBJECTIVE: The preoperative diagnosis of a primary appendiceal tumor can be difficult because of its rarity and the paucity of material in the radiology literature. The purpose of this study was to describe CT findings in six patients with primary mucinous cystadenocarcinoma of the appendix. CONCLUSION: When CT reveals a cystic mass with enhancing wall nodularity in the expected area of the appendix, especially in older patients, the possibility of primary mucinous cystadenocarcinoma should be considered.

Aged↗

The effect of continuous ambulatory peritoneal dialysis on change in serum leptin.

OBJECTIVE: Elevated serum leptin can contribute to anorexia and poor nutrition in patients with chronic renal failure, because leptin is elevated in chronic renal failure patients with or without dialysis, especially in chronic ambulatory peritoneal dialysis (CAPD) patients. The aim of this study was to find whether leptin can be removed by peritoneal dialysis (PD) and to analyze factors that can affect serum leptin after start of CAPD by observing the change in serum leptin shortly after start of CAPD and its correlation with body mass index (BMI), with serum insulin, and with residual renal function. DESIGN: Twenty patients who started CAPD during the observation period were studied. Serum leptin was measured by radioimmunoassay before start of CAPD, 3-5 days after start of CAPD, and 1 month and 3 months after start of CAPD. Simultaneously, body weight, serum insulin, and residual renal function were measured. To compensate for the circardian rhythm of leptin, removal of leptin was assessed by measuring dialysate leptin divided by average serum leptin before and after a peritoneal equilibration test (PET). RESULTS: Leptin was eliminated by PD with a dialysate-to-serum ratio of 0.16+/-0.07, which was comparable to removal of beta2-microglobulin (0.14+/-0.06). The mean serum leptin concentrations did not decrease after 3-5 days of CAPD (8.4+/-13.1 ng/mL-->11.9+/-18.0 ng/mL) despite its removal by PD, and levels increased markedly to 189% of basal serum leptin 1 month after start of PD and to 260% of basal serum leptin 3 months after start of PD. Correlation coefficients (Spearman's rho) between change of serum leptin and change of BMI, of serum insulin, of glomerular filtration rate (average of urine creatinine clearance and urine urea clearance) were 0.267 (p > 0.05, n = 20), 0.441 (p > 0.05, n = 16), 0.706 (p > 0.05, n = 8) respectively. CONCLUSION: Leptin is removed by peritoneal dialysis. Serum leptin did not decrease in 5 days after the start of PD despite its removal by PD, but increased markedly thereafter, within 3 months after start of PD. We could not find a significant correlation between the change in leptin and the change in BMI. Factors other than fat-mass gain can stimulate leptin increase shortly after start of PD.

Adipose Tissue↗

The effect of dialysate dwell on gastric emptying time in patients on continuous ambulatory peritoneal dialysis.

METHODS AND PATIENTS: We evaluated gastric emptying time (GET) with a technetium (Tc) 99m-sulfur colloid gastric emptying scan in 11 patients on continuous ambulatory peritoneal dialysis (CAPD) (6 males, 5 females) and in 14 controls. We investigated the effect of dialysate dwell on GET by studying the subjects twice: once without dialysate in the abdomen (drained) and once with 2 L of dialysate in the abdomen (full). We also investigated the relationship between body surface area (BSA) and delayed gastric emptying. RESULTS: (1) The mean gastric emptying rate in 120 minutes in patients on CAPD when drained (67.8%+/-13.4%) was not different from that in controls (65.4%+/-8.6%). (2) The mean gastric emptying rate in 120 minutes in patients on CAPD when full was significantly slower than that when drained (55.6%+/-14.6% versus 67.8%+/-13.4%, p < 0.05). In four of the 11 patients (36.4%), gastric emptying was extremely delayed from normal to abnormal range when full. (3) The BSA of patients who had extremely delayed GET from normal to abnormal range was smaller than that of patients who had minimal delayed or unchanged GET when full (1.5+/-0.11 m2 versus 1.74+/-0.22 m2). CONCLUSION: This study showed that patients on CAPD had normal gastric emptying when drained, and that gastric emptying was delayed by dialysate dwell, especially in patients who has less than 1.5 m2 of body surface area. Therefore, we suggest that, based on adequacy, intermittent nocturnal peritoneal dialysis or a small volume of dialysate be considered for patients with small body surface area.

Dialysis Solutions↗