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Biomedical subjects

B J Lee

Publications and source records attributed to B J Lee.

At least 127 records · Page 7Linked to original sources

The RNA component of mitochondrial ribonuclease P from Aspergillus nidulans.

Several RNA molecules that copurified with Aspergillus nidulans mitochondrial ribonuclease (RNase) P were identified [Lee, Y C., Lee, B. J., Hwang, D. S. & Kang, H. S. (1996) Eur J. Biochem. 235, 289-296], and their partial sequences were determined. Using an oligonucleotide probe, we cloned and mapped the gene encoding this putative RNA component of RNase P (RNase P-RNA), situated between URFA3 (unidentified reading frame A3) and cobA (apocytochrome b) genes in the mitochondrial genome of A. nidulans. The gene is extremely (A+T)-rich and contains two regions of sequence similarity conserved among the known mitochondrial RNase P-RNAs and the eubacterial RNase P-RNAs. The determination of 5' and 3' termini by primer extension and sequencing indicated that the length of the RNA transcript is 232 nucleotides. Northern-blot analysis revealed that its only subcellular location was the mitochondria. Two RNase P-RNA fragments of 110 nucleotides and 80 nucleotides, each containing one of the two conserved regions, could be recovered from the nuclease-treated enzyme without significant loss of activity. The sizes of these fragments appeared to be the minimum lengths required for the vitro activity of the enzyme.

Aspergillus nidulans↗

Conformational change of cyclic AMP receptor protein by the binding of cyclic nucleotide.

The cAMP Receptor Protein (CRP) of Escherichia coli requires a conformational change induced by the binding of cAMP in order to function as a site-specific DNA-binding protein. An intrinsic fluorescence study showed that the tryptophan residues at position 13 and 85 within CRP were located in an internal, nonpolar environment, and the conformational change induced by the binding of cAMP occurred around the tryptophan residue NMR experiment has manifested that the comformational change around the tryptophan residue at position 85 and histidine residue at position 159 of CRP is induced by the binding of cAMP. An extrinsic fluorescence study showed that the distance between the two cysteine 178 residues in the dimer was within about 3.5 A. This distance didn't exhibit a large change upon cAMP binding.

Cyclic AMP↗

Hemorrhagic fever with renal syndrome: first imported case of hantavirus infection in Taiwan.

A patient infected with hantavirus, with resultant hemorrhagic fever with renal syndrome (HFRS), is reported. The patient was a 38-year-old man, living in Tainan, Taiwan, who had been visiting Mainland China for 3 months when he suddenly developed fever and chills, generalized myalgia, abdominal pain and petechiae on his chest. He sought treatment in Mainland China. His clinical course progressed through febrile, hypotensive, oliguria and polyuria phases. Supportive care included one course of hemodialysis. He returned to Tainan in partial defervescence. Serologic studies undertaken in Taiwan confirmed Hantaan virus infection, with one of the currently identified hantavirus strains. Hospitalization with supportive care produced further clinical improvement. Clinicians should be alert to the possibility of HFRS when examining patients who have been in endemic areas and complain of fever associated with renal dysfunction, hemorrhage or abdominal pain or both.

Adult↗

Multidisciplinary evaluation of preschool children in a military psychiatry clinic: a 10-year comparison and a brief outcome study.

A little over a decade ago, the Child and Adolescent Psychiatry Service at Tripler Army Medical Center opened the Child Study Group (CSG), a multidisciplinary diagnostic team clinic for preschool children. The methods and results of the clinic were reported previously (Lee, 1987). The current study is a comparison of the cohort of children seen from January through December 1984 with the parallel group seen from January through June 1994. The paper also presents a brief follow-up study of the children seen in 1994.

Child Psychiatry↗

Molecular cloning of cDNAs encoding precursors of frog skin antimicrobial peptides from Rana rugosa.

Gaegurins, a family of peptide antibiotics with sizes ranging from 24 to 37 amino acids, have recently been purified from Rana rugosa skin (Park, J.M., Jung, J.-E. and Lee, B.J. (1994) Biochem. Biophys. Res. Commun. 205, 948-954). Two complete cDNAs encoding gaegurins 4 and 5 were isolated from a library constructed with the frog skin mRNAs. Each clone contained a single open reading frame that encodes a gaegurin precursor polypeptide. The deduced amino acid sequences revealed that the precursors have a unique tripartite structure: a putative signal sequence at the NH2-terminus followed by an acidic spacer region rich in glutamic and aspartic acids, and a mature gaegurin peptide at the COOH-terminus. Similar modes of organization were also found in antimicrobial or opioid peptide precursors of other frog species, although their mature peptides show little sequence homology. The family of peptides with this characteristic now expands. Northern analysis revealed that gaegurins are extensively expressed in the skin tissue, but not in liver and muscle.

Amino Acid Sequence↗

A monoclonal antibody reacting with the cell envelope of spirochaetes isolated from cases of intestinal spirochaetosis in pigs and humans.

A monoclonal antibody (mAb) designed BJL/AC1 was prepared against the cell envelope of an intestinal spirochaete (strain 3295) that was isolated from a pig with intestinal spirochaetosis. The mAb reacted with a band of approximately 29 kDa in cell envelope preparations from 13 porcine and 11 human spirochaetes isolated from cases of intestinal spirochaetosis, but did not react with preparations made from a range of other intestinal spirochaetes. Immunogold labelling demonstrated that the reactive epitope was located on the cell envelope of the strains causing intestinal spirochaetosis. The mAb was used in an indirect immunofluorescence test to detect spirochaetes in the faeces of pigs with experimentally induced intestinal spirochaetosis. The mAb should prove to be a useful reagent for detection and identification of spirochaetes that are specifically associated with intestinal spirochaetosis.

Animals↗

Upstream promoter elements are sufficient for selenocysteine tRNA[Ser]Sec gene transcription and to determine the transcription start point.

The TATA box, located upstream at about nt -30, and the proximal sequence element, located at about nt -60, are both essential and sufficient for basal level transcription of the Xenopus laevis (Xl) selenocysteine (Sec) tRNA[Ser]Sec gene as demonstrated by its microinjection into Xl oocytes. Point mutations within either of these regions abolish transcription, while deletion of the internal boxA element or insertion of 13 nt within the internal boxB element does not impair transcription. The latter mutations (within the internal regions) affect processing of the 3'-trailer sequence. Replacement of the tRNA[Ser]Sec coding sequence with an Escherichia coli M1 RNA gene resulted in expression of the E. coli gene governed by the upstream tRNA[Ser]Sec promoter elements. These studies demonstrate unequivocally that the upstream promoter elements are sufficient for the basal level of tRNA[Ser]Sec gene transcription. Primer extension studies with spacer mutants show that the internal elements do not play a role in selecting the transcription start point (tsp), but that selection of the tsp is determined by the region upstream from the gene. Further, studies with spacer mutants show that the distance between the TATA box and the tsp is quite likely the critical factor in selecting the position of tsp.

Animals↗

The Aspergillus uvsH gene encodes a product homologous to yeast RAD18 and Neurospora UVS-2.

The uvsH DNA repair gene of Aspergillus nidulans has been cloned by complementation of the uvsH77 mutation with a cosmid library containing genomic DNA inserts from a wild-type strain. Methylmethane sulfonate (MMS)-resistant transformants were obtained on medium containing 0.01% MMS, to which uvsH mutants exhibit high sensitivity. Retransformation of uvsH77 mutants with the rescued cosmids from the MMS-resistant transformants resulted in restoration of both UV and MMS resistance to wild-type levels. Nucleotide sequence analysis of the genomic DNA and cDNA of the uvsH gene shows that it has an open reading frame (ORF) of 1329 bp, interrupted by two introns of 51 and 61 bp. A 2.4 kb transcript of the uvsH gene was detected by Northern blot analysis. Primer extension analysis revealed that transcription starts at 31 bp upstream from the translation initiation codon. This gene encodes a predicted polypeptide of 443 amino acids, which has two unique zinc finger motifs. The proposed polypeptide displays 39% identity to the Neurospora crassa UVS-2 protein and 24% identity to the Saccharomyces cerevisiae RAD18 protein. The sequence similarity is particularly high in three domains. One zinc finger (RING finger) motif is located in the first domain close to the N-terminus. The other zinc finger motif is in the second domain. In the third domain, the mutation sites in both the uvsH77 and uvsH304 alleles were identified.(ABSTRACT TRUNCATED AT 250 WORDS)

Alleles↗

Implicating adhesion molecules in nasal allergic inflammation.

Allergic rhinitis is now considered an inflammatory disorder where many leukocyte types, including eosinophils and T-lymphocytes, accumulate in increased numbers. Along with mast cells and other cells, they release a wide variety of mediators, cytokines, and granule constituents that can directly cause inflammation or activate the local vascular endothelium to further enhance the recruitment of leukocytes through the expression and function of adhesion molecules. While the understanding of the importance of leukocyte and endothelial adhesion molecules is still at a very early stage, recent evidence has already begun to implicate these cell surface molecules in the pathogenesis of allergic diseases such as rhinitis and asthma. Additional studies, including the use of adhesion molecule antagonists when available, will clarify the importance of these structures in the pathophysiology of these disorders.

Cell Adhesion Molecules↗

Nuclear magnetic resonance study of complexes between CRP and its 22- and 28-base-pair operators.

The binding of the cAMP receptor protein (CRP) to the portion of the lac promoter comprising the core of the CRP recognition sequence has been investigated. The effect of the binding of CRP to the symmetrical 22- and 28-base-pair operators was investigated by 1H NMR. The binding of cAMP*CRP to the 22mer DNA did not bring about any changes in the chemical shift values of the imino proton resonances of the DNA, but did cause selective line broadening of the imino proton resonances of specific base pairs (TA 4, GC 5). These base pairs are contained in the motif 5'(TGTGA)3', which is thought to be the region crucial to interaction with the CRP. The binding of cAMP*CRP to the 28mer DNA brought about large changes in the imino proton resonances that seem to be induced by DNA bending. Therefore it seems likely that CRP requires DNA longer than a 22mer to bend it. We also used a C-terminal protease-digested CRP (CRPcy) in a DNA-binding experiment. The binding of cAMP*CRPcy to the 28mer DNA did not induce bending. These results indicate that the C-terminal region of CRP participates in DNA binding and is important for DNA bending.

Base Composition↗

Nodular cystic fat necrosis in a patient with erythema nodosum.

Nodular cystic fat necrosis shows a distinctive spectrum of clinical and histological features. Most of the lesions are mobile subcutaneous nodules in regions vulnerable to trauma, such as the elbows, knees, and shins. There have been no reports of an association with other conditions. The histology is characterized by encapsulated fat necrosis in which multiple, non-viable adipocytes are surrounded by condensed fibrous tissue. We report a case of early stage nodular cystic fat necrosis associated with erythema nodosum.

Adipose Tissue↗

Changes in mRNA levels of a pituitary-specific trans-acting factor, Pit-1, and prolactin during the rat estrous cycle.

The present study examined the changes in mRNA levels of a pituitary-specific trans-acting factor, Pit-1, and prolactin during the rat estrous cycle. Total cytoplasmic RNA was analyzed by Northern blot and slot-blot hybridization to examine the prolactin mRNA level. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to examine the Pit-1 mRNA level. Proestrous and estrous prolactin mRNA levels were significantly higher than the metestrous and diestrous levels, whereas Pit-1 mRNA levels of the estrous and metestrous stages were about two- to threefold higher than those of the proestrous and diestrous stages. Proestrous Pit-1 mRNA levels increased gradually from 10.00 h to 20.00 h, while prolactin mRNA levels slightly decreased until 14.00 h but increased later until 20.00 h. During the rat estrous cycle, especially in the afternoon of the proestrous day, changes of prolactin mRNA levels seem to follow a prior increase of Pit-1 mRNA. Therefore, Pit-1 may be partly involved in the regulation of prolactin gene expression according to the rat estrous cycle. Estradiol administration to ovariectomized rats significantly increased both the mRNA levels of prolactin and Pit-1, which suggests that the gene expression of Pit-1 is regulated by estrogen through indirect extracellular pathways.

Animals↗

Identification of new selenocysteine tRNA[SER]SEC isoacceptors in human cell lines.

The selenocysteine tRNA population was examined in a human T-cell line and in a human monocytic cell line for the occurrence of additional species of selenocysteine tRNA. At least three additional (and possibly more) selenocysteine isoacceptors were found which occur in minor levels as compared to the two major selenocysteine isoacceptors previously characterized. The possible significance of these newly observed species are discussed.

Cell Line↗

Antimicrobial peptides from the skin of a Korean frog, Rana rugosa.

Six antimicrobial peptides, named gaegurins, were isolated from the skin of a Korean frog, Rana rugosa, and their amino acid sequences were determined by automated Edman degradation. All peptides contain two invariant cysteine residues, one at their C-terminus and the second at the seventh position from the C-terminus. The heptapeptides containing these two cysteine residues, which we designate 'Rana boxes', are conserved in the antimicrobial peptides derived from other Rana species. Each peptide manifested a broad spectrum of antimicrobial activity against Gram positive and Gram negative bacteria, fungi and protozoa with slightly different specific activities. All gaegurins manifest very little or no hemolytic activity. These properties provide the potential for application of these peptides to effective therapeutic agents for control of pathogenic microorganisms.

Amino Acid Sequence↗

Utilization of selenocysteyl-tRNA[Ser]Sec and seryl-tRNA[Ser]Sec in protein synthesis.

The UGA selenocysteine (Sec) codon in glutathione peroxidase mRNA and in selenoprotein P and the UGA stop codon in rabbit beta-globin mRNA were employed to study the utilization of Sec-tRNA[Ser]Sec and Ser-tRNA[Ser]Sec in protein synthesis. In vitro Ser-tRNA[Ser]Sec served as a suppressor of the UGA Sec codon as well as the UGA stop codon, while Sec-tRNA[Ser]Sec did not. However, in vivo Sec-tRNA[Ser]Sec did donate Sec to glutathione peroxidase in Xenopus oocytes microinjected with glutathione peroxidase mRNA and Sec-tRNA. A ribosome binding assay was devised to investigate the interaction of aminoacyl-tRNA, rabbit reticulocyte ribosomes, and eukaryotic elongation factor 1 (eEF-1) in response to the appropriate trinucleoside diphosphate template. Ser-tRNA[Ser]Sec bound weakly to ribosomes in the presence of eEF-1 and UGA as compared to Phe-tRNA, Ser-tRNAIGA, and Met-tRNAm which bound more efficiently in the presence of eEF-1 and the appropriate template. No increase in the binding of Sec-tRNA[Ser]Sec was observed under the same conditions as Ser-tRNA[Ser]Sec. The ribosome binding studies substantiated the finding that Ser-tRNA[Ser]Sec serves as a suppressor of UGA codons in protein synthesis, but Sec-tRNA[Ser]Sec does not. In addition, these studies provide strong evidence that a specific elongation factor is required in mammalian cells for insertion of Sec into protein from Sec-tRNA[Ser]Sec.

Acylation↗

Presence of gonadotropin-releasing hormone mRNA in the rat olfactory piriform cortex.

Gonadotropin releasing hormone (GnRH) neurons are known to be originated from the olfactory placode and to enter the forebrain regions during embryonic development. The present study aims to ascertain whether GnRH is locally expressed in the olfactory cortex. Northern blot hybridization and reverse transcription-polymerase chain reaction revealed that GnRH mRNA was present in the rat olfactory cortex as well as in the olfactory bulb. The predicted size of GnRH mRNA is similar to that detected in the hypothalamus. In situ hybridization histochemistry also showed that GnRH mRNA is highly concentrated in the olfactory piriform cortex. The present data indicate that GnRH is synthesized in the olfactory piriform cortex.

Animals↗