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Biomedical subjects

H K Song

Publications and source records attributed to H K Song.

At least 91 records · Page 5Linked to original sources

Dorsal dermis of the scaleless (sc/sc) embryo directs normal feather pattern formation until day 8 of development.

We have examined the ability of the scaleless (sc/sc) backskin dermis (6 to 16 days of incubation) to regulate pattern formation using the presumptive scutate scale epidermis from 11-day normal embryos as the responding tissue. Prior to 8 days of incubation the sc/sc backskin dermis is able to induce hexagonally patterned and uniformly oriented feather germs in normal epidermis. This ability is lost during day 8 and follows a central to lateral gradient. Such gradients are characteristic of normal feather development in the spinal tract. We discuss the change in the inductive ability of the sc/sc dermis in relation to the stabilization of the feather pattern, which occurs all at once throughout the dorsal dermis at 7.5-8 days of development. After day 8 until day 10, the sc/sc backskin dermis only supports the formation of sporadic, unpatterned feather germs; thereafter it will not support feather formation.

Animals↗

Fast 3D large-angle spin-echo imaging (3D FLASE).

A rapid steady-state 3D spin-echo imaging pulse sequence, based on the principle of nutating the spins by an angle greater than 90 degrees, has been designed and implemented on a clinical 1.5-T whole-body MR scanner. The pulse sequence, denoted fast large-angle spin-echo (FLASE), has been optimized for high-resolution imaging of tissues with short T2 and T2*. Features of FLASE include a minimum-phase Shinnar-Le Roux excitation pulse and distribution of phase- and slice-encoding gradients before and after the 180 degrees refocusing pulse to minimize the critical time delay between inversion and restoration of the residual longitudinal magnetization and for minimizing echo time. A Bloch equation analysis, corroborated by experimental data, shows FLASE signal-to-noise to be superior to its closest analog, 3D rapid spin-echo excitation (RASEE) (Jara et al., Magn Reson Medicine 29, 528 (1993)), and 3D gradient-recalled acquisition in steady state (GRASS). It is demonstrated that with judicious RF phase-cycling and steady state operation, FLASE can produce high-quality microimages free of intravoxel phase dispersion from susceptibility-induced background gradients. The performance of the method is exemplified with ultra high-resolution images of trabecular bone in vitro and in vivo in the human calcaneus and wrist at voxel sizes as low as 98 x 98 x 200 microns3. Finally, the contrast behavior of refocused FLASE can be altered by disrupting the steady state analogous to gradient echo imaging.

Bone and Bones↗

Crystal structure analyses of uncomplexed ecotin in two crystal forms: implications for its function and stability.

Ecotin, a homodimeric protein composed of 142 residue subunits, is a novel serine protease inhibitor present in Escherichia coli. Its thermostability and acid stability, as well as broad specificity toward proteases, make it an interesting protein for structural characterization. Its structure in the uncomplexed state, determined for two different crystalline environments, allows a structural comparison of the free inhibitor with that in complex with trypsin. Although there is no gross structural rearrangement of ecotin when binding trypsin, the loops involved in binding trypsin show relatively large shifts in atomic positions. The inherent flexibility of the loops and the highly nonglobular shape are the two features essential for its inhibitory function. An insight into the understanding of the structural basis of thermostability and acid stability of ecotin is also provided by the present structure.

Amino Acid Sequence↗

Reverse transcriptase-polymerase chain reaction for prostate-specific antigen may be a prognostic indicator in breast cancer.

Among women with node-negative breast cancer and small tumours, it is important to identify those with tumours that will recur, so that they may receive adjuvant therapy, while sparing those with tumours that will not recur the hazards of adjuvant treatment. A reverse transcriptase-polymerase chain reaction (RT-PCR) for prostate-specific antigen (PSA) may be used to identify circulating metastatic cells in patients with prostate cancer. Approximately 30% of breast cancer cells also produce PSA. Therefore, we tested the PSA RT-PCR assay on blood specimens from women with breast cancer. We evaluated 78 women at Mount Sinai Medical Center with histologically confirmed breast cancer. Venous blood (5 cm3) from the women was collected in ethylene diaminetetraacetic acid (EDTA)-treated collection tubes and approximately 400 ng of RNA from each sample was subjected to an RT-PCR. We were able to detect the amplified PSA fragment in 18 of 78 women with breast cancer; 7 of the 18 women with the PSA fragment had localised, small, node-negative tumours, both oestrogen receptor (ER) positive and ER negative. We could not detect the amplified PSA fragment in 20 normal women and 22 normal men. We conclude that PSA RT-PCR may be a useful method for determining the presence of circulating metastatic cells in some women with node-negative breast cancer, and therefore the potential for these women to develop recurrent disease and thus benefit from adjuvant therapy.

Adult↗

In-vitro and in-vivo antibacterial activity of LB10517, a novel catechol-substituted cephalosporin with a broad antibacterial spectrum.

LB10517 is a new injectable cephalosporin with a broad spectrum of antibacterial activity against Gram-positive and Gram-negative bacteria. LB10517 inhibited 90% of methicillin-susceptible Staphylococcus aureus (MSSA) at 0.25 mg/L (MIC90), and was 8-fold more active than cefpirome. LB10517 was two- or four-fold more active than cefpirome against methicillin-susceptible Staphylococcus epidermidis (MSSE), Streptococcus pyogenes and Enterococcus faecalis. Both methicillin-resistant S. aureus (MRSA) and methicillin-resistant S. epidermidis (MRSE) were highly resistant to all compounds. LB10517 activity against most Enterobacteriaceae was comparable to or greater than that of cefpirome, and it also showed high activity against Pseudomonas aeruginosa. In a mouse septicaemia model, LB10517 exhibited excellent protective effects. In a respiratory tract infection model, the protective effect of LB10517 was comparable to that of cefpirome and ceftazidime. It was highly stable to hydrolysis by beta-lactamases, showing better physiological efficiency for TEM-9 than the other test compounds. LB10517 had the most potent antibacterial activity against beta-lactamase producing resistant strains. LB10517 did not induce beta-lactamase production in Enterobacter cloacae 1194E.

Animals↗

Crystal structure of an uncleaved alpha 1-antitrypsin reveals the conformation of its inhibitory reactive loop.

The crystal structure of a recombinant human alpha 1-antitrypsin, in the uncleaved and uncomplexed state, has been determined by X-ray crystallographic methods and refined to an R-factor of 18.4% for 8.0-3.46 A data with good stereochemistry. This structure provides the first view at the inhibitory loop and the central beta-sheet A of the uncleaved alpha 1-antitrypsin. The reactive loop takes a distorted helical conformation and no pre-insertion of two residues in the reactive loop into the beta-sheet A is observed. The present structure is largely in agreement with the model predicted by Engh, Wright, and Huber [Prot. Eng. 3 (1990) 469-477].

Binding Sites↗

Crystallization and preliminary X-ray crystallographic analysis of DNA polymerase from Thermus aquaticus.

Two crystal forms of DNA polymerase from Thermus aquaticus have been grown at room temperature. Rhombohedral crystals (form I) grown from ammonium sulfate solution diffracted poorly to 10 A only and thus are not suitable for X-ray structure determination. Trigonal crystals (form II) grown from polyethylene glycol solution are more suitable for structure determination since their diffraction pattern extends to 2.5 A at cryogenic temperature upon exposure to synchrotron X-rays. They belong to space group P3(1)21 (or its enantiomorph P3(2)21) and their unit-cell dimensions are a = 106.7 and c = 169.7 A, for flash-frozen crystals. The presence of one molecule per asymmetric unit gives a crystal volume per protein mass (V(M)) of 3.0 A(3) Da(-l) and a solvent content of 58% by volume. X-ray data have been collected to 2.7 A Bragg spacing from native crystals.

Journal Article↗

Crystallization, molecular replacement solution, and refinement of tetrameric beta-amylase from sweet potato.

Sweet potato beta-amylase is a tetramer of identical subunits, which are arranged to exhibit 222 molecular symmetry. Its subunit consists of 498 amino acid residues (Mr 55,880). It has been crystallized at room temperature using polyethylene glycol 1500 as precipitant. The crystals, growing to dimensions of 0.4 mm x 0.4 mm x 1.0 mm within 2 weeks, belong to the tetragonal space group P4(2)2(1)2 with unit cell dimensions of a = b = 129.63 A and c = 68.42 A. The asymmetric unit contains 1 subunit of beta-amylase, with a crystal volume per protein mass (VM) of 2.57 A3/Da and a solvent content of 52% by volume. The three-dimensional structure of the tetrameric beta-amylase from sweet potato has been determined by molecular replacement methods using the monomeric structure of soybean enzyme as the starting model. The refined subunit model contains 3,863 nonhydrogen protein atoms (488 amino acid residues) and 319 water oxygen atoms. The current R-value is 20.3% for data in the resolution range of 8-2.3 A (with 2 sigma cut-off) with good stereochemistry. The subunit structure of sweet potato beta-amylase (crystallized in the absence of alpha-cyclodextrin) is very similar to that of soybean beta-amylase (complexed with alpha-cyclodextrin). The root-mean-square (RMS) difference for 487 equivalent C alpha atoms of the two beta-amylases is 0.96 A. Each subunit of sweet potato beta-amylase is composed of a large (alpha/beta)8 core domain, a small one made up of three long loops [L3 (residues 91-150), L4 (residues 183-258), and L5 (residues 300-327)], and a long C-terminal loop formed by residues 445-493. Conserved Glu 187, believed to play an important role in catalysis, is located at the cleft between the (alpha/beta)8 barrel core and a small domain made up of three long loops (L3, L4, and L5). Conserved Cys 96, important in the inactivation of enzyme activity by sulfhydryl reagents, is located at the entrance of the (alpha/beta)8 barrel.

Amino Acid Sequence↗

Crystallization and preliminary X-ray crystallographic study of ribosome-inactivating protein from barley seeds.

Ribosome-inactivating protein from barley seeds has been crystallized using polyethylene glycol as precipitant. The crystal belongs to the monoclinic space group C2, with unit-cell parameters a = 88.36, b = 62.59, c = 53.18 A and beta = 108.62 degrees. The asymmetric unit contains one molecule of ribosome-inactivating protein with a corresponding crystal volume per protein mass (V(m)) of 2.32 A(3) Da(-1) and a solvent content of 47% by volume. The crystal diffracts to about 2.3 A with X-rays from a rotating-anode source and is very stable in the X-ray beam. X-ray data (nearly complete to 2.4 A Bragg spacing) have been collected from a native crystal.

Journal Article↗

Pattern formation in chick feather development: distribution of beta 1-integrin in normal and scaleless embryos.

We have examined the immunolocalization of beta 1-integrin during feather development in the spino-lumbar tract of the backskin from normal and scaleless chick embryos. beta 1-integrin appears during early feather development in three distinct phases which correspond to important developmental events. The first phase (5-5 1/2 days of incubation; Hamburger and Hamilton [H.H.] stage 27) represents the period prior to the formation of dermis. During this phase, beta 1-integrin antiserum labels mesenchymal cells located in the central region of the spino-lumbar tract where the initiation site for feather development is located. The second phase (5 1/2-7 1/2 days of incubation; H.H. stages 28-32) corresponds to the period during which dermis is formed. The cells that make up the dermis are readily distinguished by their lack of beta 1-integrin immunostaining. The third phase (7 1/2-10 days of incubation; H.H. stages 33-36) begins with the sudden appearance of beta 1-integrin in the central and lateral regions of the dermis. The pattern of beta 1-integrin immunostaining in scaleless backskin becomes different from that of normal backskin during this phase. In normal backskin the dermal condensations of feather germs are not labeled with the beta 1-integrin antiserum. This produces a heterogeneous immunostaining pattern very similar to the pattern seen for Type I collagen (Mauger et al. [1982] Dev. Biol. 94:93-105). In contrast, homogeneous immunostaining is observed in the dermis of scaleless backskin. The initial time of appearance, manner of appearance, and pattern of integrin expression in the third phase suggest that beta 1-integrin may be involved in the stabilization of the feather pattern. We also observed the appearance of beta 1-integrin on the epidermal basal cells during the time of feather follicle formation. The beta 1-integrin antiserum reacts strongly with the baso-lateral surfaces of normal basal cells, yet the basal surfaces of the scaleless basal cells are unstained. This lack of immunostaining along the basal surfaces of the scaleless basal cells may relate to the abnormal adhesion between the epidermis and dermis in scaleless backskin.

Animals↗

Histopathologic changes in the olfactory epithelium in mice after exposure to sulfur dioxide.

To investigate the effects of sulfur dioxide (SO2) on olfactory epithelium, an experiment was performed with 56 mice from the same colony. Experimental animals were divided into three groups consisting of a 30-min exposure group (group 1), a 60-min exposure group (group 2), and a 120-min exposure group (group 3). The olfactory mucosa in these mice were studied by light microscopy immediately, and after 24 h, 48 h, or 72 h exposure to 20 ppm of SO2. Edema, loss of cilia, epithelial thinning, and epithelial desquamation in the olfactory epithelium were observed in groups 2 and 3. The basal lamina and the connective tissue were well preserved throughout the entire mucosa. Injuries to olfactory epithelium became severer with exposure time. These changes were further pronounced 24 h after exposure. Regenerated epithelia were not observed in any group. Scanning electron microscopic findings were consistent with light microscopic findings. Olfactory epithelial surface were consistent with light microscopic findings. Olfactory epithelial surface was sloughed off and revealed, underlining intact basal lamina. The results of this study suggest that early lesions of olfactory epithelium after exposure to SO2 may be primarily degenerative.

Administration, Inhalation↗

Crystallization and preliminary X-ray crystallographic analysis of chitinase from barley seeds.

Chitinase from barley seeds has been crystallized at room temperature using polyethylene glycol as precipitant. The crystal is monoclinic, belonging to the space group P2(1), with unit cell parameters of a = 69.43 A, b = 44.55 A, c = 81.41 A, and beta = 111.95 degrees. The asymmetric unit seems to contain two molecules of chitinase with a corresponding crystal volume per protein mass (VM) of 2.25 A3/Da and a solvent content of 45% by volume. The crystal diffracts to at least 2.0 A with X-rays from a rotating anode source and is very stable in the X-ray beam. X-ray data have been collected to better than 2.2 A Bragg spacing from a native crystal.

Chitinases↗

Evidence that anti-CD8 abrogates anti-CD4-mediated clonal anergy but allows allograft survival in mice.

Monoclonal antibodies directed against different T cell subpopulations have been used in several rodent models of transplantation to induce long-term unresponsiveness to allografts by a variety of mechanisms. To investigate whether different mechanisms may be operative when different regimens of mAb therapy are used, we studied the effects of various combinations of anti-T-cell antibody treatment on the induction of tolerance in a mouse islet allograft model. Anti-CD4 mAb alone, anti-CD8 mAb alone, anti-CD4 mAb plus anti-CD8 mAb, and anti-Thy1.2 mAb alone were given at the time of engraftment. Only the anti-CD4 mAb and the anti-CD4 mAb plus anti-CD8 mAb regimens were successful in inducing permanent unresponsiveness to islet allografts. We have previously shown that anti-CD4 mAb alone induces permanent unresponsiveness to islet allografts by a mechanism of clonal anergy, as demonstrated by unresponsiveness of potentially alloreactive T cells to anti-T-cell receptor-specific cross-linking. Interestingly, the potentially alloreactive T cell subsets of recipient mice (V beta 5+ and V beta 11+) made unresponsive to islet allografts by anti-CD4 mAb plus anti-CD8 mAb therapy were not found to be anergic using the same assay. Differences between the repopulation kinetics of CD8+ T cells of anti-CD4 mAb plus anti-CD8 mAb treated recipient mice, which accepted islet allografts, and anti-Thy1.2 treated recipient mice, which rejected islet allografts despite similar levels of initial T cell depletion, suggest that unresponsiveness to alloantigen may have been induced in anti-CD4 mAb plus anti-CD8 mAb treated recipients by clearance of donor passenger leukocytes during prolonged CD8+ T cell depletion.

Animals↗

Evidence that clonal anergy is induced in thymic migrant cells after anti-CD4-mediated transplantation tolerance.

Diabetic (B6) (IE-) mice treated with a depleting regimen of anti-CD4 monoclonal antibody at the time of transplantation with A/J (IEK) islets of Langerhans showed indefinite acceptance of their islet allograft, as evidenced by persistent normoglycemia. To address the mechanisms involved in such anti-CD4 induced transplantation tolerance we studied potentially IE-reactive V beta 11+ T cells from the tolerant allografted mice. Following complete repopulation of the CD4+ cells, both the CD4+V beta 11+ and CD8+V beta 11+ T cell subsets of the transplanted mice were unresponsive to anti-V beta 11 specific crosslinking. In contrast, lymphocytes tested within the first ten days following transplant were responsive to anti-V beta 11 specific crosslinking; this response decreased as a function of time and reached background levels by day 120 posttransplant. Sorting experiments indicated that the response of lymphocytes to anti-V beta 11 specific crosslinking seen during the initial 120 days posttransplant was confined to the peripheral CD8+ cells; the repopulating CD4+V beta 11+ T cells were unresponsive. In addition, administration of r-IL-2 at the time of transplantation induced rejection in anti-CD4-treated animals, again indicating that the peripheral CD8+ cells could respond shortly after transplant if provided with appropriate help. The decreasing response of CD8+ T cells from transplanted animals to anti-V beta 11 stimulation was inversely correlated with the rate of migration of cells from the thymus to the periphery, implying that new thymic migrant V beta 11+ cells, both CD4+ and CD8+, were rendered anergic upon encountering peripheral alloantigen. These data suggest the possibility that recent thymic migrants are rendered anergic upon encountering antigen in the periphery, a simple model to serve as a "fail-safe" mechanism to prevent autoreactivity.

Animals↗

Past pregnancy is associated with axillary node involvement in women with breast cancer.

In 186 women with breast cancer, there was a progressive increase in the proportion of axillary nodal involvement as the number of pregnancies increased from zero to two or more (P = 0.026). Logistic regression analysis demonstrated that this effect was independent of the known relationship of age and tumor size to nodal involvement. Race and history of breast feeding had no influence on nodal involvement.

Axilla↗

Diminished ratio of estrogen receptors to progesterone receptors in breast carcinomas of women who have had multiple miscarriages.

We examined the relationship of a history of miscarriage to the ratio of tumor estrogen receptors to progesterone receptors in 74 women with breast carcinoma evaluated between 1988 and 1990. Only women who had been pregnant and whose tumor contained both estrogen and progesterone receptors were included. Women with breast carcinoma and a history of two to four miscarriages had significantly lower ratios (less than 1) of tumor estrogen receptors to progesterone receptors than women with a history of zero or one miscarriage (greater than 1). This finding may be the result of a genetic anomaly in breast cancer patients that also affects the uterine environment during the reproductive years.

Abortion, Habitual↗