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Biomedical subjects

C Kang

Publications and source records attributed to C Kang.

At least 55 records · Page 3Linked to original sources
PubMed1999

The effect of region of interest selection on dual energy X-ray absorptiometry measurements of the calcaneus in 55 post-menopausal women.

Bone mineral density (BMD) of the calcaneus was assessed by dual energy X-ray absorptiometry (DXA) in four different regions of the calcaneus in 20 pre-menopausal and 55 post-menopausal women, none of whom were on treatment or suffering from conditions affecting bone. The total body option in the small animal software package of a Lunar DPX-L bone densitometer was used. The precision of the technique (%CV) varied from 0.7 to 2.2% depending on the region scanned. For post-menopausal women, BMD results in the mid and posterior parts of the calcaneus varied by < 7% while BMD in the anterior region was about 20% lower. DXA of the calcaneus was compared with measurements at the spine (L2-L4) and hip (femoral neck) and changes with age were estimated from cross-sectional data. BMD of the calcaneus was significantly reduced in 28 post-menopausal women with low lumbar spine BMD (-2SD) compared with women with normal spine BMD. Calcaneal BMD was significantly correlated to axial BMD (r = 0.45-0.77) and to age (r = 0.45 to -0.63). For a subgroup of 33 post-menopausal women measured twice after approximately 1 year, calcaneus BMD decreased by between 1.2% and 2.5% while axial BMD showed no significant change. Unlike spine or femoral neck BMD, the decrease in calcaneus BMD was significantly greater in women with low spine BMD than in normal women, possibly indicating improved detection of skeletal changes. The optimum measurement sites for BMD in the calcaneus were within the mid or posterior part of the calcaneus or enclosing the whole posterior calcaneus. The calcaneus was shown to be a precise, sensitive and simple measurement site suitable for the assessment of osteoporosis, especially in the elderly where degenerative changes in the spine and hip can complicate BMD assessment.

Absorptiometry, Photon↗
PubMed1998

Requirement of heavy neurofilament subunit in the development of axons with large calibers.

Neurofilaments (NFs) are prominent components of large myelinated axons. Previous studies have suggested that NF number as well as the phosphorylation state of the COOH-terminal tail of the heavy neurofilament (NF-H) subunit are major determinants of axonal caliber. We created NF-H knockout mice to assess the contribution of NF-H to the development of axon size as well as its effect on the amounts of low and mid-sized NF subunits (NF-L and NF-M respectively). Surprisingly, we found that NF-L levels were reduced only slightly whereas NF-M and tubulin proteins were unchanged in NF-H-null mice. However, the calibers of both large and small diameter myelinated axons were diminished in NF-H-null mice despite the fact that these mice showed only a slight decrease in NF density and that filaments in the mutant were most frequently spaced at the same interfilament distance found in control. Significantly, large diameter axons failed to develop in both the central and peripheral nervous systems. These results demonstrate directly that unlike losing the NF-L or NF-M subunits, loss of NF-H has only a slight effect on NF number in axons. Yet NF-H plays a major role in the development of large diameter axons.

Actin Cytoskeleton↗
PubMed1998

Probing RegA/RNA interactions using electrospray ionization-fourier transform ion cyclotron resonance-mass spectrometry.

The interactions of bacteriophage T4 regA protein, a unique translational regulator, with RNAs of various size and sequence were studied using electrospray ionization-Fourier transform ion cyclotron resonance-mass spectrometry. Using very gentle interface conditions, regA/RNA complexes with a 1:1 binding stoichiometry were observed for all four target RNAs studied, consistent with solution binding studies. Competitive binding of target RNAs and their degradation products with regA demonstrated that the loss of a single nucleotide resulted in a dramatic change in binding affinity in some cases. Competitive binding of regA with four target RNAs revealed similar relative binding affinity order to that suggested by previous in vitro repression experiments. The use of sustained off-resonance irradiation for collisionally induced dissociation of a regA/RNA complex suggested the potential for directly obtaining information regarding the regA binding domain.

Bacterial Proteins↗
PubMed1998

Absence of the mid-sized neurofilament subunit decreases axonal calibers, levels of light neurofilament (NF-L), and neurofilament content.

Neurofilaments (NFs) are prominent components of large myelinated axons and probably the most abundant of neuronal intermediate filament proteins. Here we show that mice with a null mutation in the mid-sized NF (NF-M) subunit have dramatically decreased levels of light NF (NF-L) and increased levels of heavy NF (NF-H). The calibers of both large and small diameter axons in the central and peripheral nervous systems are diminished. Axons of mutant animals contain fewer neurofilaments and increased numbers of microtubules. Yet the mice lack any overt behavioral phenotype or gross structural defects in the nervous system. These studies suggest that the NF-M subunit is a major regulator of the level of NF-L and that its presence is required to achieve maximal axonal diameter in all size classes of myelinated axons.

Animals↗
PubMed1998

Phylogenetic analysis of the nef gene reveals a distinctive monophyletic clade in Korean HIV-1 cases.

To study the genetic variation of the HIV-1 strains prevalent in South Korea, we analyzed the nef sequences derived from 46 HIV-1-positive individuals living in various geographic regions in Korea. Phylogenetic analysis revealed four subtypes of HIV-1: A (3 patients), B (41 patients), D (1 patient), and a type that could not be clearly classified to any known subtype (1 patient). Thirty-five of the 41 Korean subtype B isolates formed a distinct monophyletic clade that is not related to any of the international sequences from the Los Alamos Database or GenBank as of June 1997. Indeed, the presence of unique conserved sequences was identified among the Korean isolates in this Korean subtype B group. The variations in the nucleotide sequences of a majority (32 of 35) subtype B samples within the Korean clade were 1.9% to 8.8%, and amino acid sequences varied from 3.9% to 15.5%. These results suggest that HIV-1 strains currently present in South Korea might have originated from a few sources or might be developing through a certain selective pressure. This is the first report on the molecular nature of the HIV-1 infection present in South Korea.

Adolescent↗
PubMed1998

Effects of cigarette smoking and 12-h abstention on working memory during a serial-probe recognition task.

Nicotine has been shown to affect attentional and mnemonic processes. However, whether these effects are due to changes in perceptual and/or motor aspects of the tasks is not at all clear. This study tested the hypothesis that nicotine from cigarette smoking has differential effects on perceptual and motor processes, as reflected by event-related potentials (ERPs) and reaction times (RTs), respectively, and that perceptual effects may be specific to changes in working memory. ERPs. RTs and performance accuracy were recorded from smokers and nonsmokers during a serial-probe recognition memory task in which lists of words or "memory sets" were followed by a probe word that was either in-set or out-of-set. Smokers were tested in a "smoking" and a 12-h "deprived" condition. Smoking-smokers and deprived-smokers exhibited fast RTs to in-set and out-of-set probes relative to a group of nonsmokers. They exhibited even faster RTs when the inset probe word matched the first or last item in the memory set. Thus, smokers as a group showed enhanced primacy and recency effects suggesting that smoking specifically facilitates processes related to the motor output aspects of working memory. Different effects characterized the electrophysiology. Larger P300s were recorded to in-set compared to out-of-set probes by both subject groups. Smoking smokers exhibited enhanced P300s to both types of probes. When smokers abstained for 12 h (deprived smokers), the differences in P300 amplitude were reduced but not eliminated. Smoking smokers exhibited faster P300 latencies to in-set probes, while deprived smokers showed delayed latencies relative to nonsmokers. Primacy and recency P300 effects characterized nonsmokers and deprived smokers. However, this relationship was reversed in the Smoking condition. These results support the hypotheses that nicotine has distinct effects on memory-related perceptual and motor aspects of working memory. The increase in efficiency of the memory search with nicotine is consistent with the functional role of the cholinergic system in maintaining a state "appropriate for efficient information processing."

Adolescent↗
PubMed1998

Viability of E. coli cells containing phage RNA polymerase and promoter: interference of plasmid replication by transcription.

Strong transcription of phage promoters often renders the host E. coli cells containing the phage RNA polymerase inviable. When expression of the phage SP6 RNA polymerase gene in one plasmid was induced in the E. coli JM109 cells, cells that bear an active SP6 promoter were inviable. When it was not induced (the polymerase was still produced in low levels), viability of the host cells and stability of the promoter-bearing plasmids depended on the orientation of the promoter with respect to that of the replication origin and on the sequence of the origin. A group of SP6 promoter-bearing plasmids (group I plasmids) that had the promoter directed towards the ColE1 replication origin, rendered the polymerase-containing host cells inviable in selective media. When the sequence of the origin was different (group II plasmids), this adverse effect was not observed. When the promoter direction was same as the replication origin and the ampicillin-resistant gene (group III plasmids), many satellites formed around the colonies on ampicillin-containing agar plates. These effects were caused by strong transcription of the phage SP6 promoter by its RNA polymerase, since they were reduced or eliminated by inserting an active terminator just downstream of the promoter. The viability of host cells and copy number of the promoter/terminator-bearing plasmids appear to be quantitatively related with efficiency of initiation and termination of the phage transcription. These systems may be useful for in vivo screening for mutant variants of the phage promoter, polymerase and terminator that are affected in their efficiency.

Bacteriophages↗
PubMed1998

Development and differentiation of the anuran auditory brainstem during metamorphosis: an acetylcholinesterase histochemical study.

The time course of cell differentiation and the presence of histochemically defined areas in brainstem auditory nuclei were examined in developing bullfrogs, Rana catesbeiana, using cresyl violet staining and acetylcholinesterase (AChE) histochemistry. In the medulla, the dorsolateral nucleus (DLN) can be seen as a distinct structure in its adult location only at Gosner stage 40 and beyond. The majority of cells in the DLN are not fully differentiated until late metamorphic climax (stages 45-46) and early postmetamorphosis. The more ventral vestibular nucleus differentiates earlier (stage 37) than the DLN. Adult-like organization of auditory nuclei in the torus semicircularis (TS) of the midbrain cannot be reliably discerned until metamorphic climax stages. Cellular masses in the brainstem reveal AChE from the earliest stage examined (stage 27) but the intensity of staining differs among cell groups. Staining intensity in the DLN is at a peak in recently metamorphosed froglets. The time course of cell differentiation in the DLN precedes slightly or is coincident with the increased, transient presence of AChE. Staining of the superior olive stabilizes at a moderate level in early postmetamorphic stages. Ventral regions of the principal nucleus in the TS stain more intensely than dorsal regions beginning at stage 40. This dorsal-ventral gradient in staining persists in adult stages. There is a transient decline in staining of the laminar nucleus in metamorphic climax stages. Staining intensity in the magnocellular nucleus peaks during stages 40-46 and in early postmetamorphic froglets and then declines in adults, paralleling the pattern seen in the DLN. These data suggest that metamorphic climax and early froglet periods are an important developmental window for major differentiation and maturational events in the auditory brainstem.

Acetylcholinesterase↗
PubMed1998

Comparison of ultrasound and dual energy X-ray absorptiometry measurements in the calcaneus.

Ultrasound measurements have been proposed as a means of providing structural information on bone but have also been shown to correlate with density. High correlation coefficients have been obtained in vitro, but are lower for in vivo work. The aim of this study was to investigate the relationship of broadband ultrasound attenuation (BUA), speed of sound (SOS) and stiffness, to bone mineral density (BMD) measured in the calcaneus at a closely matched region of interest (ROI). Measurements were made in 55 post-menopausal and 20 young, normal women. Calcaneus BMD measurements were made using an ROI (fixed by reference to external axes) to represent the area and location of the ultrasound transducers and an ROI based on anatomical markers, positioned in the posterior part of the calcaneus. BUA was significantly correlated with calcaneus BMD in the fixed ROI (r = 0.77, p < 0.0001) and the anatomical ROI (r = 0.78, p < 0.0001). Correlation of BUA with axial BMD was lower at r = 0.63, p < 0.0001 for the spine (L2-L4) and r = 0.59, p < 0.0001 for the femoral neck. Results for SOS and stiffness were very similar. From the calcaneus dual energy X-ray absorptiometry (DXA) scan images the region representing the ultrasound measurement area was found to contain cortical bone edges in 60% of cases. In 16% of scans a small part (4.5 +/- 4.0%) of the ROI measurement area fell partially outside the heel. However, there was no obvious discrepancy in the ultrasound results in the individual cases where this occurred. The correlation between calcaneus BMD and BUA was improved from r = 0.72 to r = 0.86 when scans with a cortical edge in the measurement ROI were excluded. The values for SOS were similar. In summary, BMD of the calcaneus, when closely matched to the site of ultrasound measurement shows significant correlation with ultrasound measurements. By excluding scans in which the ultrasound measurement appeared to include bone edges, correlations of approximately r = 0.86 were obtained. However, the BMD result still does not explain all of the variance in the ultrasound results.

Absorptiometry, Photon↗
PubMed1998

[Study on the power of O/E method].

This paper addresses the methodology of calculating the power of O/E method and introduces the relevant algorithm for estimating the sample size. Based on the principles of hyporthesis testing and Poisson distribution, the power can be worked out for given sample size, baseline rate(pi 0) and the factor of increasing (gamma). The power increases with the increase in sample size, pi 0 and gamma. In shape the power is not a continuous smooth line but a zigzag line. The method illustrated in this paper can be also used for estimaging the sample size of studies on the intervention of diseases.

Congenital Abnormalities↗
PubMed1998

[Surgical treatment of epispadias].

OBJECTIVE: To analyse the surgical treatment of 124 cases of epispadias (incomplete 27 cases, complete 21 and complex 76). METHOD: Leadbetter's operation was used for anti-incontinence. Urethral tightening was used in 4 cases of complete female epispadias. Penile straightening was adopted for incomplete epispadias and penile lengthening for complete and complex male epispadias. RESULT: In 97 cases of urinary incontinence, complete continence was achieved in 73. CONCLUSION: The orthopedic effects of external genitals were satisfactory.

Adolescent↗
PubMed1997

A common core for binding single-stranded DNA: structural comparison of the single-stranded DNA-binding proteins (SSB) from E. coli and human mitochondria.

The crystal structure of the DNA-binding domain of E. coli SSB (EcoSSB) has been determined to a resolution of 2.5 A. This is the first reported structure of a prokaryotic SSB. The structure of the DNA-binding domain of the E. coli protein is compared to that of the human mitochondrial SSB (HsmtSSB). In spite of the relatively low sequence identity between them, the two proteins display a high degree of structural similarity. EcoSSB crystallises with two dimers in the asymmetric unit, unlike HsmtSSB which contains only a dimer. This is probably a consequence of the different polypeptide chain lengths in the EcoSSB heterotetramer. Crucial differences in the dimer-dimer interface of EcoSSB may account for the inability of EcoSSB and HsmtSSB to form cross-species heterotetramers, in contrast to many bacterial SSBs.

Crystallography, X-Ray↗
PubMed1997

Single-stranded RNA recognition by the bacteriophage T4 translational repressor, regA.

The T4 protein, RegA, is a translational repressor that blocks ribosome binding to multiple T4 messages by interacting with the mRNAs near their respective AUG start codons. Other than the AUG, there are no obvious similarities between the affected mRNAs. High affinity RNA ligands to RegA were isolated using SELEX (systematic evolution of ligands by exponential enrichment). The selected RNAs exhibited the consensus sequence 5'-AAAAUUGUUAUGUAA-3'. The AUG was invariant, suggesting that it is the primary effector of binding specificity. The UU immediately 5' to the AUG and the upstream poly(A) tract were highly conserved among the selected RNAs. Boundary and footprinting experiments are consistent with the consensus sequence defining the RegA-binding site. Interestingly, chemical modification and nuclease digestion data indicate that the RNA-binding site is single-stranded, as if RegA discriminates between targets based on their primary sequence, not their secondary structure. Minor variations from the consensus at positions other than the universally conserved AUG have little effect on RegA binding, but accumulation of mutations has a profound effect on the interaction. Comparison of the in vivo targets for RegA to the SELEX-generated consensus suggests a repression pattern whereby the translation of individual messages is sequentially halted until the least similarly affected message, the regA gene itself, is repressed.

Bacteriophage T4↗
PubMed1997

Residues essential for catalysis and stability of the active site of Escherichia coli adenylosuccinate synthetase as revealed by directed mutation and kinetics.

Examined here by directed mutation, circular dichroism spectroscopy, and kinetics are the relationships of five residues, Asp13, Glu14, Lys16, His41, and Arg131, to the catalytic function and structural organization of adenylosuccinate synthetase from Escherichia coli. The D13A mutant has no measurable activity. Mutants E14A and H41N exhibit 1% of the activity of the wild-type enzyme and 2-7-fold increases in the Km of substrates. The mutant K16Q has 34% of the activity of wild-type enzyme and Km values for substrates virtually unchanged from those of the wild-type system. Mutation of Arg131 to leucine caused only a 4-fold increase in the Km for aspartate relative to the wild-type enzyme. The dramatic effects of the D13A, E14A, and H41N mutations on kcat are consistent with the putative roles assigned to Asp13 (catalytic base), His41 (catalytic acid), and Glu14 (structural organization of the active site). The modest effect of the R131L mutation on the binding of aspartate is also in harmony with recent crystallographic investigations, which suggests that Arg131 stabilizes the conformation of the loop that binds the beta-carboxylate of aspartate. The modest effect of the K16Q mutation, however, contrasts with significant changes brought about by the mutation of the corresponding lysines in the P-loop of other GTP- and ATP-binding proteins. Crystallographic structures place Lys16 in a position of direct interaction with the gamma-phosphate of GTP. Furthermore, lysine is present at corresponding positions in all known sequences of adenylosuccinate synthetase. We suggest that along with a modest role in stabilizing the transition state of the phosphotransfer reaction, Lys16 may stabilize the enzyme structurally. In addition, the modest loss of catalytic activity of the K16Q mutant may confer such a selective disadvantage to E. coli that this seemingly innocuous mutation is not tolerated in nature.

Adenylosuccinate Synthase↗
PubMed1997

Crystal structure of human mitochondrial single-stranded DNA binding protein at 2.4 A resolution.

We solved the crystal structure of the homotetrameric single-stranded DNA binding (SSB) protein from human mitochondria at a resolution of 2.4 A. The tetramer is formed by two dimers interacting head-to-head and shows D2 symmetry. Sequence-related tetrameric SSB proteins occur in prokaryotes and eukaryotic mitochondria; this is the first report of an atomic resolution structure of this type of protein. Using biochemical data and analysis of sequence homologies, we were able to correlate the functional properties with structure. We propose that ssDNA wraps around the tetrameric HsmtSSB protein through electropositive channels guided by flexible loops.

Amino Acid Sequence↗
PubMed1997

Detection of resistance to isoniazid, rifampin, and streptomycin in clinical isolates of Mycobacterium tuberculosis by molecular methods.

Molecular techniques can detect drug resistance in Mycobacterium tuberculosis, but whether these methods are practical for clinical laboratory use and the management of tuberculosis is unclear. We evaluated several available molecular methods (restriction fragment length polymorphism [RFLP], heteroduplex, and direct DNA sequence analyses) for detecting resistance to isoniazid, rifampin, and streptomycin and compared these methods with conventional methods for susceptibility testing. RFLP analysis detected the mutation at position S315T in katG in 12 (44.4%) of 27 isoniazid-resistant strains. Heteroduplex analysis of rpoB, detected 16 (76.2%) of 21 rifampin-resistant strains, whereas direct DNA sequencing detected all rifampin-resistant strains. RFLP analysis of the rpsL gene detected only nine (28.1%) of 32 streptomycin-resistant strains, while direct DNA sequencing detected nearly 68% of streptomycin-resistant strains. The specificity of all of the above-mentioned methods was excellent. RFLP analysis for katG and direct DNA sequencing of rpoB and rpsL may be practical methods for routine use in clinical microbiology laboratories or molecular pathology laboratories with good molecular capabilities and autosequencers. Despite the less than optimal sensitivity for some assays, resistance can be detected rapidly. However, molecular methods are not yet capable of replacing more traditional methods of susceptibility testing for M. tuberculosis.

Antibiotics, Antitubercular↗
PubMed1997

Lipoprotein(a) isoforms display differences in affinity for plasminogen-like binding to human mononuclear cells.

Binding of lipoprotein(a) (Lp(a)) to membrane proteins of the monocyte-macrophage cell lineage may be an important event in atheroma formation. Since Lp(a) with distinct apolipoprotein(a) (apo(a)) isoforms may show differences in their affinity with regard to fibrin binding, the existence of such a functional behavior in the interaction of apo(a) in Lp(a) with these cells was explored using the monocytic cell line THP-1. Lp(a) preparations containing small size apo(a) isoforms (M(r) = 450,000 to 550,000) and high molecular mass isoforms (M(r) > or = 700,000) were purified from plasmas containing > 0.35 g/L of Lp(a) obtained from subjects (n = 14) with cardiovascular atherosclerotic disease. Binding of plasminogen to THP-1 cells was performed using the method of radioisotopic dilution. For binding of Lp(a) to cells, the THP-1 monocytic cells were incubated with varying concentrations of the different Lp(a) preparations; cells were then washed and the amount of Lp(a) bound was detected with a radiolabeled polyclonal antibody directed against apo(a). Binding due to kringle interactions with lysine residues was calculated by subtracting from the total bound the amount of Lp(a) bound (approximately 10%) in the presence of 6-aminohexanoic acid. Analysis of data with the Langmuir equation indicated identical and independent (non-interacting) sites and allowed evaluation of the Kd. Binding isotherms of small size isoforms showed saturation and a high affinity (Kd = 25.8 +/- 19 nmol/L) relative to that of plasminogen (Kd = 1750 +/- 760 nmol/L). A similar difference (Kd = 17.5 +/- 7.9 nmol/L versus Kd = 600 +/- 220 nmol/L) was found when binding experiments were performed with a fibrin surface. In contrast, binding isotherms of the high molecular mass isoforms did not show saturation at the highest Lp(a) concentrations used, thus indicating a lower affinity. In conclusion, these results show that apo(a) isoforms may display polymorphism-linked functional heterogeneity with regard to cell binding, which may explain the higher association with cardiovascular risk of small size isoforms. These qualitative differences in the binding of apo(a) isoforms to fibrin or cells may modulate the cardiovascular risk associated with high levels of Lp(a).

Adult↗