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Biomedical subjects

J M Hu

Publications and source records attributed to J M Hu.

At least 19 recordsLinked to original sources
PubMed2002

The impact of intraoperative transoesophageal echocardiography on an unselected cardiac surgical population: a review of 2343 cases.

Although intraoperative transoesophageal echocardiography (TOE) has an established role in the management of some cardiac surgical procedures, there is little data on its impact on coronary artery bypass graft (CABG) cases that are stratified for clinical risk. This is a retrospective review of the surgical impact of intraoperative TOE on 2,343 unselected cardiac cases. The surgical impact of TOE findings were rated E (essential)--resulted in changes to the proposed surgical procedure or V (valuable)--the surgical technique for the planned surgery was altered. The surgical impact that routine TOE had on low-, medium- and high-risk CABG cases was also examined. The surgical impact of TOE for the total study population (E + V) was 4.5%. The impact was greatest in combined CABG + mitral valve procedures (18%). The impact in CABG cases was 3.5% overall, with an estimated impact in low-risk patients of 2.8% (95% CI. 2.7-3.0%) versus 6.7% (95% CI. 5.9-7.7%) in high-risk cases. The commonest E-impact in CABG patients was unscheduled valve surgery (2.6% of high-risk patients). The complication rate attributable to TOE was 0.09%. These findings provide indirect evidence for a potential patient benefit from the routine use of TOE in cardiac surgery.

Aged↗
PubMed2001

[Spectral study on five complexes derived from D-glucosamine and their effect on DNA].

Complexes CuGlu, ZnGlu, NiGlu, Fe(III) Glu, Co(III) Glu where Glu is d-glucosamine have been synthesized. The effects of these compounds on DNA have been studied by the UV-Vis, fluorescence and SERS methods. The possible active ways of these complexes with DNA have been discussed. The results show that the adsorbed state of these complexes on silver sols are similar, and that CuGlu, Co(III) Glu have strong interactions with DNA. Their acting ways on DNA are different. It is worth to do a further research on these compounds as anticancer drugs.

Antineoplastic Agents↗
PubMed2000

Error, bias, and long-branch attraction in data for two chloroplast photosystem genes in seed plants.

Sequences of two chloroplast photosystem genes, psaA and psbB, together comprising about 3,500 bp, were obtained for all five major groups of extant seed plants and several outgroups among other vascular plants. Strongly supported, but significantly conflicting, phylogenetic signals were obtained in parsimony analyses from partitions of the data into first and second codon positions versus third positions. In the former, both genes agreed on a monophyletic gymnosperms, with Gnetales closely related to certain conifers. In the latter, Gnetales are inferred to be the sister group of all other seed plants, with gymnosperms paraphyletic. None of the data supported the modern "anthophyte hypothesis," which places Gnetales as the sister group of flowering plants. A series of simulation studies were undertaken to examine the error rate for parsimony inference. Three kinds of errors were examined: random error, systematic bias (both properties of finite data sets), and statistical inconsistency owing to long-branch attraction (an asymptotic property). Parsimony reconstructions were extremely biased for third-position data for psbB. Regardless of the true underlying tree, a tree in which Gnetales are sister to all other seed plants was likely to be reconstructed for these data. None of the combinations of genes or partitions permits the anthophyte tree to be reconstructed with high probability. Simulations of progressively larger data sets indicate the existence of long-branch attraction (statistical inconsistency) for third-position psbB data if either the anthophyte tree or the gymnosperm tree is correct. This is also true for the anthophyte tree using either psaA third positions or psbB first and second positions. A factor contributing to bias and inconsistency is extremely short branches at the base of the seed plant radiation, coupled with extremely high rates in Gnetales and nonseed plant outgroups.

Chloroplasts↗
PubMed2000

Effect of taurine in rat milk on the growth of offspring.

The physiological significance of taurine in milk in the growth of rat pups was investigated. Our results confirmed that taurine was at an exceptionally high concentration in rat milk during the lactational period, especially for the first few days after birth. Pups taking no milk from natural dams but from foster mothers at an advanced lactational period showed a slower growth rate. Intraperitoneal administration of taurine to the foster mothers in the first five days restored this growth retardation. On the other hand, intraperitoneal administration of beta-alanine, a transport antagonist of taurine, to the natural dams through the lactational period induced a slower growth rate of pups. This beta-alanine treatment to dams increased beta-alanine concentration, but did not decrease taurine concentrations in milk, and serum taurine concentration in the pups receiving this milk was elevated. Direct administration of beta-alanine to pups also increased the serum taurine concentrations dose-dependently. Beta-alanine administration to pups significantly decreased [3H]taurine incorporation into all the organs examined, and in contrast. [3H]taurine concentrations in serum and urine were elevated. Thus, beta-alanine inhibited taurine incorporation into cells and accelerated taurine excretion into either urine or milk. Serum IGF-I levels in pups receiving beta-alanine either directly or via their mothers was significantly lower than those in control pups. Cumulatively, taurine ingestion from milk at an early lactational period seems critical for normal growth of rat neonates due to its role in maintaining normal serum IGF-I levels.

Animals↗
PubMed2000

Expression of cysteine sulfinate decarboxylase mRNA in rat mammary gland.

Our previous report demonstrated that high concentration of taurine is present in rat milk for the first few days of lactation and plays an important role in the body growth of rat pups. In the present study, gene expression of rate-limiting enzyme for taurine biosynthesis, cysteine sulfinate decarboxylase (CSD) were examined in rat mammary gland. By RT-PCR, CSD mRNA was found to be expressed in rat mammary gland like that in the liver. The expression level of CSD mRNA in the mammary gland was higher in the earlier lactational stage (days 1 and 6 of lactation) than that in the later lactational stage (day 14). CSD mRNA expression in the mammary gland of non-pregnant rats was only a trace level. By in situ hybridization analysis, CSD mRNA was demonstrated in the epithelial cells of the mammary gland. These results suggest that high concentrations of taurine in the milk are at least partially resulted from de novo synthesis of taurine in mammary gland epithelial cells and that the expression pattern of CSD mRNA may be responsible for the changes in taurine levels in the milk during a lactational period.

Animals↗
PubMed2000

Phylogenetic systematics of the tribe Millettieae (Leguminosae) based on chloroplast trnK/matK sequences and its implications for evolutionary patterns in Papilionoideae.

Phylogenetic relationships in the tribe Millettieae and allies in the subfamily Papilionoideae (Leguminosae) were reconstructed from chloroplast trnK/matK sequences. Sixty-two accessions representing 57 traditionally recognized genera of Papilionoideae were sampled, including 27 samples from Millettieae. Phylogenies were constructed using maximum parsimony and are well resolved and supported by high bootstrap values. A well-supported "core Millettieae" clade is recognized, comprising the four large genera Millettia, Lonchocarpus, Derris, and Tephrosia. Several other small genera of Millettieae are not in the core Millettieae clade. Platycyamus is grouped with Phaseoleae (in part). Ostryocarpus, Austrosteenisia, and Dalbergiella are neither in the core Millettieae or Phaseoleae clade. These taxa, along with core Millettieae and Phaseoleae, form a monophyletic sister group to Indigofereae. Cyclolobium and Poecilanthe are close to Brongniartieae. Callerya and Wisteria belong to a large clade that includes all the legumes that lack the inverted repeat in their chloroplast genome, which confirms previous rbcL and phytochrome gene family phylogenies. The evolutionary history of four characters was examined in Millettieae and allies: the presence of canavanine, inflorescence types, the dehiscence of pods, and the presence of winged pods. trnK/matK sequence analysis suggests that the presence of a pseudoraceme or pseudopanicle and the accumulation of nonprotein amino acids are phylogenetically informative for Millettieae and allies with only a few exceptions.

Journal Article↗
PubMed1998

Isolation and characterization of plasmid pSW200 from Erwinia stewartii.

The nucleotide sequence of pSW200 of Erwinia stewartii SW2 was determined. This plasmid is 4367 bp long, consisting of four mobilization genes, mobCABD, and an origin of replication homologous to those of ColE1-type plasmids. The plasmid also contains a region of forty-one 15-bp repeats. Deleting this region does not affect the stability or the copy number when maintained as sole plasmid in the cell. However, the plasmid is rapidly lost when a homoplasmid with the intact repeat region is introduced into the cell. The function of this region may provide pSW200 an advantage in competing with an incompatible plasmid in the cell.

Amino Acid Sequence↗
PubMed1998

Glucocorticoid receptor phosphorylation: overview, function and cell cycle-dependence.

All steroid hormone receptors are phosphorylated and undergo hormone-induced hyperphosphorylation. Most phosphorylated residues identified so far are serines in the N-terminal domain. Other residues and domains may also be phosphorylated, e.g. the estrogen receptor is phosphorylated on tyrosine in the hormone-binding domain. Many sites lie in consensus sequences for proline-directed, cell cycle-associated kinases. In some receptors hyperphosphorylation is induced by hormone antagonists as well as agonists, and leads to new phosphorylated sites. With glucocorticoid receptors, hyperphosphorylation is specific for glucocorticoid agonists, follows receptor activation and produces no new sites. Rate studies suggest that hyperphosphorylation is due to accelerated phosphorylation rather than delayed dephosphorylation. Evidence to date indicates that steroid hormone receptor phosphorylation serves not as an on-off switch but modulates function more subtly. Mutations of phosphorylated sites to alanine have been found to decrease activity by 0 to 90%, depending on mutated site, cell type, reporter gene and hormone concentration. With glucocorticoid receptors, some alanine mutants are up to 75% less active in hormone-induced transactivation of certain reporter genes. They are also inactive in hormone-induced repression of transcription of their own gene and down regulation of the receptor protein. Furthermore, they are much less sensitive to degradation. Both basal phosphorylation and hormone-dependent hyperphosphorylation of these receptors are cell cycle-dependent, basal phosphorylation being low in S phase and high in G2/M and hyperphosphorylation the reverse, suggesting a causal relation to the cell cycle-dependence of glucocorticoid activity reported with several cell lines. Hyperphosphorylation appears to be regulated by basal phosphorylation through negative charge in the N-terminal domain, which in S phase is relatively low and permits hyperphosphorylation, but in G2/M is relatively high and blocks hyperphosphorylation.

Cell Cycle↗
PubMed1997

Control by basal phosphorylation of cell cycle-dependent, hormone-induced glucocorticoid receptor hyperphosphorylation.

Mouse glucocorticoid receptors (GRs) are phosphorylated in the N-terminal domain at serine/ threonine residues, most lying in consensus sequences for cell cycle-associated kinases. Glucocorticoid agonists, but not antagonists, induce hyperphosphorylation. Phosphorylation of GRs overexpressed in Chinese hamster ovary (CHO) cells is cell cycle-dependent: basal phosphorylation in S phase is one third that in G2/M; glucocorticoids induce hyperphosphorylation in S but not G2/M, paralleling the reported sensitivity in S and resistance in G2/M of proliferating cells to transcriptional activation by glucocorticoids. This parallel led us to investigate what controls hyperphosphorylation. We tested three hypotheses: hyperphosphorylation is controlled by 1) negative charge due to basal GR phosphorylation, being permitted in S by low charge and blocked in G2/M by high charge; 2) presence in S and absence in G2/M of required kinases; 3) availability in S and lack in G2/M of unoccupied phosphorylatable sites. Our results are inconsistent with 2) and 3), but strongly support 1). GR mutants with alanines (A7GR) or glutamates (E7GR) replacing all but one phosphorylated site were overexpressed in CHO cells. Serine 122 remained intact to report GR phosphorylation. Consistent with hypothesis 1, with A7GRs hormone-induced hyperphosphorylation occurred in both S and G2/M (thus revealing kinase activity for hyperphosphorylation of at least serine 122 in both phases), whereas with E7GRs it occurred in neither phase. We conclude that basal GR phosphorylation controls hormone-induced GR hyperphosphorylation by modulating negative charge in the N-terminal domain and could potentially control other cell cycle-dependent GR properties.

Animals↗
PubMed1995

Hormone-induced hyperphosphorylation of specific phosphorylated sites in the mouse glucocorticoid receptor.

The glucocorticoid receptor (GR) is phosphorylated in its basal state, and rapidly undergoes hormone-induced hyperphosphorylation after binding glucocorticoids. Previously, we have identified seven phosphorylated sites in the mouse GR. Most of the sites are located in the regions of the N-terminal domain that are necessary for maximum transcriptional activity and reduce nonspecific binding to DNA. Using WCL2 cells, which overexpress mouse GRs, we now quantitate hormone-induced hyperphosphorylation at each of these sites. Addition of triamcinolone acetonide to WCL2 cells results in significant hyperphosphorylation at the majority of the sites. The hyperphosphorylation ratio, i.e. the 32P incorporation into GRs from hormone-treated cells divided by 32P incorporation into GRs from untreated cells, was above 1.0 for all sites but serine 150 and threonine 159. Serine 220 displays marked hormone dependence, with a ratio of 3. For most sites the ratio was about 1.5. Hormone-induced hyperphosphorylation not only increases the charge at selected phosphorylated sites but also provides a substantial increase in the overall negative charge around the region of the N-terminal domain that is involved in transactivation.

Animals↗
PubMed1994

Glucocorticoid receptors in ATP-depleted cells. Dephosphorylation, loss of hormone binding, HSP90 dissociation, and ATP-dependent cycling.

Dependence of hormone binding to glucocorticoid receptors (GRs) on cellular ATP levels suggested that GRs traverse an ATP-dependent cycle, and without ATP accumulate in forms that cannot bind hormone. Such "null" receptors (NRs) were identified in ATP-depleted WEHI-7 cells, where they are tightly associated with the nuclear fraction and partly dephosphorylated. With WCL2 cells (Chinese hamster ovary cells with overexpressed GRs) depleted of ATP with azide, we have now identified dephosphorylated sites on NRs, studied possible roles of phosphorylation using GR mutants, and measured association with the 90-kDa heat shock protein (hsp90). Most NRs in WCL2 cells are dephosphorylated at serines 220 and 234, but GRs with those serines mutated to alanines do not resemble NRs since they bind hormone. They do not associate strongly with nuclei. On azide treatment, however, mutated GRs lose hormone binding capacity faster than normal GRs. Association of hsp90 (and presumably other heat shock proteins) with cytosolic GRs is drastically reduced by azide treatment, sufficient to account for decreased hormone binding. We conclude that: (a) dephosphorylation of GRs does not yield NRs, but may weaken association with hsp90. (b) The postulated ATP-dependent GR cycle can be accounted for by dissociation, and ATP-dependent reconstitution, of GR-hsp90 complexes. (c) ATP depletion blocks reconstitution of complexes. Uncomplexed GRs may accumulate as one form of NR; they are probably also the precursors for other forms of NR.

Adenosine Triphosphate↗
PubMed1994

Cell cycle-dependent glucocorticoid receptor phosphorylation and activity.

Proliferating cells display striking cell cycle dependence in sensitivity to gene activation by glucocorticoids; they are sensitive in late gap 1/synthesis (G1/S) (late G1 and S phases) but resistant in gap 2/mitotic (G2/M). Here we describe large cell cycle-dependent variations in glucocorticoid receptor (GR) phosphorylation that accompany, and may account for, the changes in sensitivity. GRs are basally phosphorylated and undergo hyperphosphorylation after hormone-induced activation. Identified phosphorylated sites are all in the N-terminal domain. Several lie in a region required for full transactivating activity and reduction of nonspecific binding to DNA. Most are in consensus sequences for cell cycle-associated kinases, suggesting that such kinases phosphorylate GRs. We now show with WCL2 cells (Chinese hamster ovary cells with overexpressed GRs) that: 1) glucocorticoid treatment fails to hyperphosphorylate GRs in G2/M but doubles phosphorylation in S, more than seen with unsynchronized cells; and 2) basal GR phosphorylation is almost three times higher in G2/M than S. These results, along with earlier observations, implicate GR phosphorylation with mechanisms of glucocorticoid resistance in G2/M. Such mechanisms might underlie some forms of glucocorticoid resistance in inflammatory and lymphoproliferative diseases. HPLC phosphopeptide maps of GRs from S and G2/M reveal no significant qualitative differences in phosphorylated sites, consistent with a general increase during G2/M in negative charge of the N-terminal domain. We also show that the previously described increase in GR hormone-binding capacity from G1 to S is accompanied by a parallel increase in GR protein.

Animals↗
PubMed1993

Glucocorticoid receptors: ATP-dependent cycling and hormone-dependent hyperphosphorylation.

The dependence of hormone binding to glucocorticoid receptors (GRs) on cellular ATP levels led us to propose that GRs normally traverse an ATP-dependent cycle, possibly involving receptor phosphorylation, and that without ATP they accumulate in a form that cannot bind hormone. We identified such a form, the null receptor, in ATP-depleted cells. GRs are basally phosphorylated, and become hyperphosphorylated after treatment with hormone (but not RU486). In mouse receptors we have identified 7 phosphorylated sites, all in the N-terminal domain. Most are on serines and lie within a transactivation region. The time-course of hormone-induced hyperphosphorylation indicates that the primary substrates for hyperphosphorylation are the activated receptors; unliganded and hormone-liganded nonactivated receptors become hyperphosphorylated more slowly. After dissociation of substrates for hyperphosphorylation are the activated receptors; unliganded and hormone-liganded nonactivated receptors become hyperphosphorylated more slowly. After dissociation of hormone, most receptors appear to be recycled and reutilized in hyperphosphorylated form. From these and related observations, we have concluded that the postulated ATP-dependent cycle can be accounted for by hormone-induced or spontaneous dissociation of receptor-Hsp90 complexes, followed by reassociation of unliganded receptors with Hsp90 via an ATP-dependent reaction like that demonstrated in cell-free systems. Other steroid hormone receptors might traverse a similar cycle. Four of the 7 phosphorylated sites in the N-terminal domain are in consensus sequences for p34cdc2 kinases important in cell cycle regulation. This observation, along with the known cell cycle-dependence of sensitivity to glucocorticoids and other evidence, point to a role for receptor phosphorylation in controlling responses to glucocorticoids through the cell cycle.

Adenosine Triphosphate↗
PubMed1992

Functional analyses of albumin expression in a series of hepatocyte cell lines and in primary hepatocytes.

A series of simian virus 40-immortalized hepatocyte cell lines which are heterogeneous with regard to expression of albumin protein and RNA were characterized for their ability to transcribe the albumin gene. Nascent chain extension assay showed that albumin RNA levels in these cells were determined predominantly at the transcription level. The albumin promoter and enhancer sequences were fused to the bacterial chloramphenicol acetyltransferase gene; the ability of the resulting expression constructs to drive chloramphenicol acetyltransferase expression after transfection into these hepatocyte cell lines was measured. The activity of the albumin promoter and enhancer constructs in primary hepatocytes was also measured. The albumin promoter was expressed differentially in these cells; however, no correlation was found between the transcriptional efficiency of the transfected albumin promoter and endogenous albumin transcription. The albumin enhancer was functional in some but not all albumin-positive cells. The minimal albumin enhancer was mapped to a 330-base pair fragment extending from -9.94 kilobases (kb) to -10.27 kb; three elements within this fragment recently shown to be necessary for enhancer function in a murine hepatocyte cell line were also essential for albumin enhancer function in the rat hepatocyte cell line CWSV1. A transcriptional silencer was identified which could suppress the expression of the homologous albumin promoter and the heterologous herpes simplex virus thymidine kinase promoter. Preliminary analysis localized the albumin silencer between -11 and -12 kb. Our results suggest that multiple regulatory sequences may act cooperatively to determine efficient tissue-specific expression of the albumin gene.

Albumins↗
PubMed1991

Differing activities of medullary respiratory neurons in eupnea and gasping.

Our purpose was to compare further eupneic ventilatory activity with that of gasping. Decerebrate, paralyzed, and ventilated cats were used; the vagi were sectioned within the thorax caudal to the laryngeal branches. Activities of the phrenic nerve and medullary respiratory neurons were recorded. Antidromic invasion was used to define bulbospinal, laryngeal, or not antidromically activated units. The ventilatory pattern was reversibly altered to gasping by exposure to 1% carbon monoxide in air. In eupnea, activities of inspiratory neurons commenced at various times during inspiration, and for most the discharge frequency gradually increased. In gasping, the peak discharge frequency of inspiratory neurons was unaltered. However, all commenced activities at the start of the phrenic burst and reached peak discharge almost immediately. The discharge frequencies of all groups of expiratory neurons fell in gasping, with many neurons ceasing activity entirely. These data are consistent with the hypothesis that brain stem mechanisms controlling eupnea and gasping differ fundamentally.

Animals↗
PubMed1989

Activities of two new antiviral agents against guinea pig lymphotropic herpesvirus infection in vitro.

Two new antiviral agents, compound 164, also known as 2'-nor-cGMP (9-[(2-hydroxy-1,3,2-dioxaphosphorinan-5-yl)oxymethyl]-guani ne P-oxide), and compound 102 [4-amino-5-bromo-7-(2-hydroxyethoxymethyl)-pyrrolo(2,3-d)pyrimidine], together with acyclovir for comparison, were evaluated for activities against the guinea pig lymphotropic herpesvirus infection in vitro by plaque reduction and virus yield reduction assays in guinea pig embryo cells. The two new compounds were demonstrated to be more potent against guinea pig lymphotropic herpesvirus infections than acyclovir. Compound 164 was the most potent of the three; drug concentrations required to reduce the number of plaques by 50% were 2, 35.5, and 144.5 microM for compounds 164, 102, and acyclovir, respectively. The two new compounds were cytostatic but not cytotoxic to guinea pig embryo cells in cultures. Attempts were made to investigate the inhibition of viral replication by these compounds, and the influence of test conditions on antiviral evaluations is discussed.

Acyclovir↗
PubMed1989

[Studies on the origin of different discharge pattern of the two types of phrenic motoneurones by cross-correlation analysis].

Experiments were performed on twenty five anesthetized, paralyzed, vagotomized and artificially ventilated rabbits. Discharges of single phrenic nerve fibers were recorded. According to their discharge patterns, phrenic motoneurones were divided into two types: tonic units (TU) and phasic units (PU). A cross-correlation analysis was done between TU and TU, PU and PU, or TU and PU. In 21/36 pairs of phrenic motoneurones, the cross-correlation histogram showed a significant central peak, which represented synchronization of the pair of units resulting from a common input. There were three types of synchronization: short-term, broad peak and high frequency-oscillatory synchronization. The difference in synchronization number between heterogeneous pairs (1 TU and 1PU, 2/10 showing synchronization) and homogeneous pairs (2 TU or 2 PU, 19/26 showing synchronization) was statistically significant (x2-test, P less than 0.05). These results suggest that TU and PU have different central inputs, which result in their different discharge patterns between the TU and PU.

Animals↗