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Biomedical subjects

B Kim

Publications and source records attributed to B Kim.

At least 145 records · Page 8Linked to original sources

Growth kinetics and treatment response of the intracerebral rat 9L brain tumor model: a quantitative in vivo study using magnetic resonance imaging.

We report the use of magnetic resonance imaging (MRI) for in situ tumor growth rate studies of experimental intracranial 9L tumors. T2-weighted spin-echo coronal magnetic resonance images of rat brains with 9L tumors were obtained every 2 days beginning at 8-11 days postimplantation using a 7 tesla MRI system. Tumors were clearly delineated in the images as a hyperintense region with a relatively well-demarcated border and minimal peritumoral edema. Tumor volumes from individual slices were summed together to yield the total tumor volume. The accuracy of this methodology for volumetric determination was verified by MRI phantom studies. Tumor growth rates determined from sequential MRI measurements of tumor volumes were quantitated in terms of volumetric doubling time. Tumor doubling times were found to range from 50 to 81 h, with an average of 66 +/- 8 h (n = 10). Intracranial 9L tumors were found to grow exponentially over the entire life span of the animal, allowing treated animals to serve as their own controls since the volumetric doubling time could be determined from three to four MRI scans before treatment administration. The intracerebral tumor growth delay following a single injection of 1, 3-bis(2-chloroethyl)-1-nitrosourea (13.3 mg/kg i.p.) allowed for noninvasive determination of in vivo log cell kill. A 2.0 +/- 0.2 (n = 3) log cell kill from 1,3-bis(2-chloroethyl)-1-nitrosourea treatment was found from post-treatment MRI volume measurements. These results demonstrate that MRI provides a powerful and sensitive method for assessing the growth and treatment response of intracranial 9L tumors in the rat.

Animals↗

Assessment of ganciclovir toxicity to experimental intracranial gliomas following recombinant adenoviral-mediated transfer of the herpes simplex virus thymidine kinase gene by magnetic resonance imaging and proton magnetic resonance spectroscopy.

Magnetic resonance imaging and in vivo localized H magnetic resonance spectroscopy were used to evaluate a gene therapy approach for treating experimental brain tumors. This approach involved the use of an adenoviral vector to transfer the herpes simplex virus thymidine kinase (HSVtk) gene into intracerebral 9L gliosarcomas in rats followed by systemic administration of the antiherpetic agent ganciclovir. Magnetic resonance imaging quantitation of changes in intracranial 9L tumor doubling times revealed a significant variation in therapeutic response. Localized H magnetic resonance spectra of 9L tumors treated with Ad.RSVtk/ganciclovir revealed a dramatic increase in the resonance intensity at 0.9-1.3 ppm, corresponding to mobile lipids and/or lactate. Changes in intracranial tumor doubling times correlated with changes in H tumor magnetic resonance spectra, suggesting that specific changes in tumor metabolite levels may be predictive of the effectiveness of this gene therapy approach.

Animals↗

DNA polymerase delta is required for base excision repair of DNA methylation damage in Saccharomyces cerevisiae.

We present evidence that DNA polymerase delta of Saccharomyces cerevisiae, an enzyme that is essential for viability and chromosomal replication, is also required for base excision repair of exogenous DNA methylation damage. The large catalytic subunit of DNA polymerase delta is encoded by the CDC2(POL3) gene. We find that the mutant allele cdc2-2 confers sensitivity to killing by methyl methanesulfonate (MMS) but allows wild-type levels of UV survival. MMS survival of haploid cdc2-2 strains is lower than wild type at the permissive growth temperature of 20 degrees C. Survival is further decreased relative to wild type by treatment with MMS at 36 degrees C, a nonpermissive temperature for growth of mutant cells. A second DNA polymerase delta allele, cdc2-1, also confers a temperature-sensitive defect in MMS survival while allowing nearly wild-type levels of UV survival. These observations provide an in vivo genetic demonstration that a specific eukaryotic DNA polymerase is required for survival of exogenous methylation damage. MMS sensitivity of a cdc2-2 mutant at 20 degrees C is complemented by expression of mammalian DNA polymerase beta, an enzyme that fills single-strand gaps in duplex DNA in vitro and whose only known catalytic activity is polymerization of deoxyribonucleotides. We conclude, therefore, that the MMS survival deficit in cdc2-2 cells is caused by failure of mutant DNA polymerase delta to fill single-strand gaps arising in base excision repair of methylation damage. We discuss our results in light of current concepts of the physiologic roles of DNA polymerases delta and epsilon in DNA replication and repair.

Alleles↗

Adapting homogeneous enzyme-linked competitive binding assays to microtiter plates.

Recently devised homogeneous enzyme-linked binding assays useful for the rapid detection of carbohydrate structure/content of intact glycoproteins (via use of lectins as binders) and for quantitating given vitamins (e.g., biotin; using soluble binding proteins) are adapted successfully to a microtiter plate reader format. The problem of nonspecific adsorption of the binders and enzyme-saccharide/vitamin conjugates is solved via the addition of Tween 20 to the assay buffer. More convenient and reliable photometric detection of the preferred labeling enzyme, glucose-6-phosphate dehydrogenase (G6PDH), is accomplished by monitoring the rate of generation of reduced thio-NAD (from thio-NAD) at 405 nm instead of NADH (from NAD) at 340 nm. By employing these modifications it is shown that homogeneous enzyme-linked binding assays can be readily adapted to microtiter plates without loss in analytical assay performance. Results further suggest that other homogeneous assays based on G6PDH, including commercial EMIT assays used routinely in clinical chemistry laboratories for detecting drugs of abuse, could, in principle, be run on microtiter plates to significantly enhance sample throughput.

Binding, Competitive↗

Bladder tumor staging: comparison of contrast-enhanced CT, T1- and T2-weighted MR imaging, dynamic gadolinium-enhanced imaging, and late gadolinium-enhanced imaging.

PURPOSE: To evaluate the accuracy of contrast material-enhanced computed tomography (CT), unenhanced T1- and T2-weighted magnetic resonance (MR) imaging, and dynamic gadolinium-enhanced spoiled gradient-echo MR imaging in the staging of bladder cancer. MATERIALS AND METHODS: Thirty-six patients with histologically proved bladder cancer were prospectively examined with CT and MR imaging before tumor resection. Images were evaluated in a blinded fashion. RESULTS: All methods depicted tumors comparably well. Staging was correct in 16 of 29 patients (55%) with CT and 27 of 36 patients (75%) overall with MR imaging. Overstaging was the most common error, and both CT and MR imaging were more accurate for higher-staged tumors. CONCLUSION: CT and MR imaging perform well in the detection of tumors but are less accurate in staging. Dynamic gadolinium-enhanced MR imaging is better than other techniques, but the difference is not statistically significant.

Carcinoma, Transitional Cell↗

Sulfation of parabens and tyrosylpeptides by bacterial arylsulfate sulfotransferases.

Arylsulfate sulfotransferase purified from Eubacterium A-44 has higher specific activity than the enzymes from Klebsiella K-36 and Haemophilus K-12. Propylparaben and butylparaben were good substrates among several parabens. The antibacterial activity of parabens was reduced by the sulfation of the phenolic hydroxy group. Tyrosine-containing peptides, kyotorphin, enkephalin and cholecystokinin non-sulfate, were effective as acceptor substrates by A-44, K-36 and K-12 sulfotransferases.

Animals↗

Pineal toxoplasmosis mimicking pineal tumor in an AIDS patient.

A pineal mass in a patient with acquired immunodeficiency syndrome (AIDS) is reported. Computed tomography (CT) scan revealed a nodular mass in the pineal region with foci of calcification and obstruction of the aqueduct mimicking a pineal tumor. At autopsy, the brain revealed a well-circumscribed lesion with central necrosis in the pineal region suggestive of toxoplasma and involving the periaqueductal area. Susceptibility of a patient with AIDS to opportunistic infections should be considered.

AIDS-Related Opportunistic Infections↗

LexA and lambda Cl repressors as enzymes: specific cleavage in an intermolecular reaction.

During the SOS response, LexA repressor is inactivated by specific cleavage. Although cleavage requires RecA protein in vivo, RecA acts indirectly as a coprotease by stimulating an inherent self-cleavage activity of LexA. In lambda lysogens, cleavage of lambda Cl repressor in a similar but far slower reaction results in prophage induction. We describe an intermolecular cleavage reaction in which the C-terminal fragment of LexA acted as an enzyme to cleave other molecules of LexA. The C-terminal fragment of lambda repressor cleaved the LexA substrates about as efficiently as did the LexA enzyme, suggesting that the slow rate of Cl self-cleavage results from a weak interaction between its cleavage site and the active site.

Bacterial Proteins↗

Expression of protein kinase C isozymes in rat glial cell line.

To study the presence and distribution of various isozymes of PKC in glial cells, immunofluorescent studies were performed on a cortical glial cell line cultured derived from rat cerebral cortex and analyzed by a highly sensitive automated laser cytometric device, ACAS-570 (Adherent Cell Analysis and Sorting). The antibodies tested were against alpha, beta, gamma and epsilon isozymes of PKC, and the catalytic domain (cd), i.e. antibody against the cd to all isozymes of PKC. The data indicate that rat cortical glial cell line expresses at least three isozymes of PKC, epsilon, beta, and alpha, the degree of intensity of immunofluorescence being epsilon > beta > alpha. The immunofluorescence for PKC-cd was the most intense.

Animals↗

[Seroepidemiological study of human rotavirus in northern part of Tokyo from 1990 to 1992 by using reverse transcription-polymerase chain reaction].

A seroepidemiological study of rotavirus was conducted in the northern part of Tokyo from 1990 to 1992 by using reverse transcription-polymerase chain reaction (RT-PCR). G1 and G3 types were detected in the winter between 1990 and 1991, however G1 type was appeared mainly in the winter between 1991 and 1992. G3 type was observed as the main type during the winter in the 10 year survey in the Tokyo area for the first time. RT-PCR was useful in the seroepidemiological studies of rotavirus infection.

Base Sequence↗

Diagnosis of urethral diverticula in women: value of MR imaging.

OBJECTIVE: The purpose of this study was to determine the role of MR imaging in the diagnosis of urethral diverticula in women. SUBJECTS AND METHODS: In 20 patients (mean age, 47 years), MR imaging was performed (1.5-T); unenhanced T1-weighted and T2-weighted images and contrast-enhanced T1-weighted images were obtained. MR imaging findings were compared with findings on urethrograms (including voiding cystourethrograms or double balloon catheter studies) in 16 patients, urethroscopic findings in 18, and surgical findings in 13. MR images were assessed for the presence, size, number, and location (anterior or posterior to the urethral lumen and relationship to the neck of the bladder) of diverticula, visualization of ostia, and presence of any concomitant complications. Sixteen of 20 patients had urethral diverticula: uncomplicated (simple) urethral diverticula in 13 patients; a diverticulum with a stone in one; a diverticulum with chronic granulation in one; and a diverticulum with adenocarcinoma in one. In the remaining four patients, the diagnoses included Gartner's duct cyst (one) and paraurethral cyst (one); urethras were deemed to be normal in two. RESULTS: In 13 patients who had surgery, MR images correctly showed the presence or absence of diverticula, whereas urethrography was correct in nine (69%) and urethroscopy in 10 (77%). Compared with surgical findings (20 diverticula in 12 patients), MR images depicted 14 (70%) of 20 diverticula and urethrography and urethroscopy each depicted 11 (55%) of 20. The ostium of the diverticulum could not be identified on MR images. The use of MR contrast material did not contribute to lesion detection, but the enhanced images were superior to unenhanced images in showing granulation tissue and carcinoma, although they did not help in their differentiation. CONCLUSION: MR imaging is accurate for showing urethral diverticula, but owing to its high cost, it should be considered only when urethroscopic or urethrographic findings are equivocal or when patients are unable to undergo these procedures and clinical findings strongly suggest a urethral diverticulum.

Adult↗