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Biomedical subjects

H Green

Publications and source records attributed to H Green.

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

Association of p63 with proliferative potential in normal and neoplastic human keratinocytes.

p63, a recently identified member of the p53 gene family, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. We show that in normal human epidermis, in hair follicles, and in stratified epidermal cultures, p63 protein is principally restricted to cells with high proliferative potential and is absent from the cells that are undergoing terminal differentiation. In normal human epidermis and in hair follicles, basal cells with abundant p63 are interspersed with cells with little or no p63. Whenever p63 mRNA is present, it encodes mainly truncated, potentially dominant-negative isotypes. In squamous cell carcinomas, the number of cells containing p63 and their distribution depends on the degree of anaplasia. In highly differentiated tumors, p63 is confined to a ring of basal-like cells surrounding, but at a distance from, centers of terminal differentiation. In less differentiated tumors, most cells contain p63 and their distribution is chaotic with respect to centers of terminal differentiation. p63 appears to be a valuable diagnostic marker for anaplastic keratinocytes.

Carcinoma, Squamous Cell↗
PubMed1999

Suppression of the biocontrol agent trichoderma harzianum by mycelium of the arbuscular mycorrhizal fungus glomus intraradices in root-free soil

Trichoderma harzianum is an effective biocontrol agent against several fungal soilborne plant pathogens. However, possible adverse effects of this fungus on arbuscular mycorrhizal fungi might be a drawback in its use in plant protection. The objective of the present work was to examine the interaction between Glomus intraradices and T. harzianum in soil. The use of a compartmented growth system with root-free soil compartments enabled us to study fungal interactions without the interfering effects of roots. Growth of the fungi was monitored by measuring hyphal length and population densities, while specific fatty acid signatures were used as indicators of living fungal biomass. Hyphal 33P transport and beta-glucuronidase (GUS) activity were used to monitor activity of G. intraradices and a GUS-transformed strain of T. harzianum, respectively. As growth and metabolism of T. harzianum are requirements for antagonism, the impact of wheat bran, added as an organic nutrient source for T. harzianum, was investigated. The presence of T. harzianum in root-free soil reduced root colonization by G. intraradices. The external hyphal length density of G. intraradices was reduced by the presence of T. harzianum in combination with wheat bran, but the living hyphal biomass, measured as the content of a membrane fatty acid, was not reduced. Hyphal 33P transport by G. intraradices also was not affected by T. harzianum. This suggests that T. harzianum exploited the dead mycelium but not the living biomass of G. intraradices. The presence of external mycelium of G. intraradices suppressed T. harzianum population development and GUS activity. Stimulation of the hyphal biomass of G. intraradices by organic amendment suggests that nutrient competition is a likely means of interaction. In conclusion, it seemed that growth of and phosphorus uptake by the external mycelium of G. intraradices were not affected by the antagonistic fungus T. harzianum; in contrast, T. harzianum was adversely affected by G. intraradices.

Journal Article↗
PubMed1999

Initial aerobic power does not alter muscle metabolic adaptations to short-term training.

To investigate the hypothesis that training-induced increases in muscle mitochondrial potential are not obligatory to metabolic adaptations observed during submaximal exercise, regardless of peak aerobic power (VO(2 peak)) of the subjects, a short-term training study was utilized. Two groups of untrained male subjects (n = 7/group), one with a high (HI) and the other with a low (LO) VO(2 peak) (means +/- SE; 51.4 +/- 0.90 vs. 41.0 +/- 1.3 ml. kg(-1). min(-1);P < 0.05), cycled for 2 h/day at 66-69% of VO(2 peak) for 6 days. Muscle tissue was extracted from vastus lateralis at 0, 3, and 30 min of standardized cycle exercise before training (0 days) and after 3 and 6 days of training and analyzed for metabolic and enzymatic changes. During exercise after 3 days of training in the combined HI + LO group, higher (P < 0.05) concentrations (mmol/kg dry wt) of phosphocreatine (40.5 +/- 3.4 vs. 52.2 +/- 4.2) and lower (P < 0.05) concentrations of P(i) (61.5 +/- 4.4 vs. 53.3 +/- 4.4), inosine monophosphate (0.520 +/- 0.19 vs. 0.151 +/- 0.05), and lactate (37.9 +/- 5.5 vs. 22.8 +/- 4.8) were observed. These changes were also accompanied by reduced levels of calculated free ADP, AMP, and P(i). All adaptations were fully expressed by 3 min of exercise and by 3 days of training and were independent of initial VO(2 peak) levels. Moreover, maximal activity of citrate synthase, a measure of mitochondrial capacity, was only increased with 6 days of training (5.71 +/- 0.29 vs. 7.18 +/- 0.37 mol. kg protein(-1). h(-1); P < 0. 05). These results demonstrate that metabolic adaptations to prolonged exercise occur within the first 3 days of training and during the non-steady-state period. Moreover, neither time course nor magnitude of metabolic adaptations appears to depend on increases in mitochondrial potential or on initial aerobic power.

Adaptation, Physiological↗
PubMed1999

Regulation of fiber size, oxidative potential, and capillarization in human muscle by resistance exercise.

To examine the hypothesis that increases in fiber cross-sectional area mediated by high-resistance training (HRT) would result in a decrease in fiber capillarization and oxidative potential, regardless of fiber type, we studied six untrained males (maximum oxygen consumption, 45.6 +/- 2.3 ml. kg-1. min-1; mean +/- SE) participating in a 12-wk program designed to produce a progressive hypertrophy of the quadriceps muscle. The training sessions, which were conducted 3 times/wk, consisted of three sets of three exercises, each performed for 6-8 repetitions maximum (RM). Measurements of fiber-type distribution obtained from tissue extracted from the vastus lateralis at 0, 4, 7, and 12 wk indicated reductions (P < 0.05) in type IIB fibers (15.1 +/- 2.1% vs. 7.2 +/- 1.3%) by 4 wk in the absence of changes in the other fiber types (types I, IIA, and IIAB). Training culminated in a 17% increase (P < 0.05) in cross-sectional area by 12 wk with initial increases observed at 4 wk. The increase was independent of fiber type-specific changes. The number of capillaries in contact with each fiber type increased by 12 wk, whereas capillary contacts-to-fiber area ratios remained unchanged. In a defined cross-sectional field, HRT also increased the capillaries per fiber at 12 wk. Training failed to alter cellular oxidative potential, as measured by succinic dehydrogenase (SDH) activity, regardless of fiber type and training duration. It is concluded that modest hypertrophy induced by HRT does not compromise cellular tissue capillarization and oxidative potential regardless of fiber type.

Adult↗
PubMed1999

Downregulation of Na+-K+-ATPase pumps in skeletal muscle with training in normobaric hypoxia.

To investigate the effects of training in normoxia vs. training in normobaric hypoxia (fraction of inspired O2 = 20.9 vs. 13.5%, respectively) on the regulation of Na+-K+-ATPase pump concentration in skeletal muscle (vastus lateralis), 9 untrained men, ranging in age from 19 to 25 yr, underwent 8 wk of cycle training. The training consisted of both prolonged and intermittent single leg exercise for both normoxia (N) and hypoxia (H) during a single session (a similar work output for each leg) and was performed 3 times/wk. Na+-K+-ATPase concentration was 326 +/- 17 (SE) pmol/g wet wt before training (Control), increased by 14% with N (371 +/- 18 pmol/g wet wt; P < 0.05), and decreased by 14% with H (282 +/- 20 pmol/g wet wt; P < 0.05). The maximal activity of citrate synthase, selected as a measure of mitochondrial potential, showed greater increases (P < 0.05) with H (1.22 +/- 0.10 mmol x h-1 x g wet wt-1; 70%; P < 0.05) than with N (0.99 +/- 0.10 mmol x h-1 x g wet wt-1; 51%; P < 0.05) compared with pretraining (0.658 +/- 0.09 mmol x h-1 x g wet wt-1). These results demonstrate that normobaric hypoxia induced during exercise training represents a potent stimulus for the upregulation in mitochondrial potential while at the same time promoting a downregulation in Na+-K+-ATPase pump expression. In contrast, normoxic training stimulates increases in both mitochondrial potential and Na+-K+-ATPase concentration.

Adult↗
PubMed1998

Inability of keratinocytes lacking their specific transglutaminase to form cross-linked envelopes: absence of envelopes as a simple diagnostic test for lamellar ichthyosis.

Epidermal keratinocytes, late in their terminal differentiation, form cross-linked envelopes resistant to ionic detergent and reducing agent. Because the cross-linking process is catalyzed by the keratinocyte transglutaminase, the absence of active transglutaminase should result in failure of the keratinocyte to form a cross-linked envelope. Three keratinocyte strains bearing mutations in the keratinocyte transglutaminase were examined: two contained no detectable transglutaminase mRNA and none contained active enzyme. All three were unable to form cross-linked envelopes, either spontaneously in stratified cultures or upon induction with Ca2+. Although stratum corneum of normal humans and scales from patients with different ichthyotic diseases contain cross-linked envelopes, those from patients with transglutaminase-negative lamellar ichthyosis do not. Therefore, the disease due to the absence of transglutaminase may be readily distinguished from other ichthyotic disease by a simple test for cross-linked envelopes.

Cell Count↗
PubMed1998

The effects of photic driving on mood states.

The EEG photic driving response is a sensitive neurophysiological measure. It has been used to assess drug effects, forms of epilepsy, neurological status of Alzheimer's patients, and physiological arousal. Photic driving also impacts the psychological status of a person by producing increased visual imagery and decreased physiological and subjective arousal. In this study, ten volunteers underwent nocturnal polysomnography followed by six daytime testing sessions. The six sessions consisted of the alpha attenuation test, two visual analog scales for mood, the Stanford Sleepiness Scale, photic stimulation, and the multiple sleep latency test. These tests were administered 2 hours upon awakening and every 2 hours thereafter. The mean mood across the six daytime testing sessions was computed for all mood variables pre- and post-photic stimulation. Significant differences were found for the subjective moods "sleepy," "alert," and "effort." However, no significant differences were found for pre- and post-photic driving for "angry," "irritable," "hungry," "tense," "overall," "happy," "sexual," and "sad." Additionally, all participants reported increased visual imagery during photic driving, as measured by their responses to an imagery questionnaire.

Adult↗
PubMed1998

Transglutaminase action imitates Huntington's disease: selective polymerization of Huntingtin containing expanded polyglutamine.

Different proteins bearing polyglutamine of excessive length are lethal to neurons and cause human disease of the central nervous system. In parts of the brain affected by Huntington's disease, the amount of the huntingtin with expanded polyglutamine is reduced and there appear huntingtin-containing polymers of larger molecular weight. We show here that huntingtin is a substrate of transglutaminase in vitro and that the rate constant of the reaction increases with length of the polyglutamine over a range of an order of magnitude. As a result, huntingtin with expanded polyglutamine is preferentially incorporated into polymers. Both disappearance of the huntingtin with expanded polyglutamine and its replacement by polymeric forms are prevented by inhibitors of transglutaminase. The effect of transglutaminase therefore duplicates the changes in the affected parts of the brain.

Adolescent↗
PubMed1998

Basonuclin as a cell marker in the formation and cycling of the murine hair follicle.

Basonuclin, a zinc-finger protein, is found in stratified squamous epithelia and hair follicles. In the basal keratinocytes of mouse epidermis, basonuclin is detected mainly in the cytoplasm. During the development of murine hair follicles, this protein concentrates in the nuclei of the basal cells that form the primary hair germs. As follicle morphogenesis proceeds, the epithelial cells possessing nuclear basonuclin invade the dermis and surround the follicular papilla. In mature anagen follicles, nuclear basonuclin is principally restricted to the basal layers of the outer root sheath and bulbar matrix; these regions are known to contain cells capable of proliferation, and to lack the features of terminal differentiation. During catagen, the compartment of cells containing nuclear basonuclin regresses, and in telogen, only a small number of these cells remain to form the secondary hair germ at the follicle base. During the next anagen, this basonuclin-containing population expands and regenerates the hair-producing portion of the follicle. It is concluded that in all hair cycles, the transient segment of the follicle originates from germinative cells possessing nuclear basonuclin.

Animals↗
PubMed1997

Conservation of human and mouse basonuclins as a guide to important features of the protein.

The nucleotide sequence of mouse basonuclin has been determined from its cDNA by PCR and compared with the previously known sequence of human basonuclin. Overall, there is 88% identity in the encoded amino acid sequences, but some regions have been much more conserved than others. Zinc fingers 2 and 6, the region containing the nuclear localization signal and the region containing the serine stripe encode identical amino acid sequences in the two species, but differ by numerous silent nucleotide substitutions, suggesting that these regions are likely to be important for the functions of the protein common to the two species. Similarly, zinc fingers 1 and 5 diverge at only a single amino acid residue. In contrast, other regions of the sequence have diverged considerably, such as zinc fingers 3 and 4. The region adjacent to the N-terminus is very divergent and this aids in locating the translation start site. The highly conserved regions are likely to be essential for the common function of the proteins, and the more divergent regions may be either unconstrained or adapted to different requirements in the two species.

Amino Acid Sequence↗
PubMed1997

Nuclear localization of basonuclin in human keratinocytes and the role of phosphorylation.

Basonuclin is a zinc-finger protein found in basal cells of the epidermis. In human keratinocyte cultures, basonuclin is susceptible to serine-phosphorylation and the addition of the phosphatase inhibitor, okadaic acid, promotes accumulation of basonuclin in the cytoplasm. The region of basonuclin containing the nuclear localization signal of basonuclin is necessary for nuclear localization of the protein and Ser-541, located immediately C-terminal to the nuclear localization signal, is the principal phosphorylation site in vitro. A nearly complete basonuclin transiently expressed in cultured keratinocytes localizes predominantly in the nucleus, but substitution of aspartic acid for Ser-541 promotes cytoplasmic localization. The same substitution of Ser-537 has a similar but weaker effect. Substitution of both serine residues by alanine leads to nuclear localization. These results show that nuclear localization of basonuclin depends on serine dephosphorylation, primarily of Ser-541. Different subcellular locations of basonuclin in different keratinocyte subtypes are therefore most likely to be controlled by the state of phosphorylation of Ser-541.

3T3 Cells↗
PubMed1997

The human basonuclin gene.

The human gene for basonuclin, a zinc-finger protein of keratinocytes, has been cloned, sequenced and assigned to chromosome 15. The transcription unit spans nearly 29 kb of sequence. The coding region is distributed over five exons, and the three pairs of zinc fingers are encoded by the last two. The 5' flanking sequence and first exon are unusually rich in G+C and in CpG dinucleotides. This region contains numerous target sites for the transcription factor Sp1.

Amino Acid Sequence↗
PubMed1997

The involucrin gene of the tree shrew: recent repeat additions and the relocation of cysteine codons.

The coding region of the involucrin gene of Tupaia glis has been cloned and sequenced. It resembles the involucrin coding region of other non-anthropoid mammals in possessing a segment of related, short tandem repeats at a defined location, but in Tupaia, there has been recent serial duplication of a repeat into which a cysteine codon had earlier been introduced. As a result of the duplication, there is a total of as many as six cysteine codons in the segment of repeats, a number larger than for any other species yet examined. In Ratttus there has been a comparable but independent addition of cysteine codons, and both Tupaia and Rattus have eliminated an otherwise conserved cysteine codon 75 located close to but outside the segment of repeats. In Tupaia, this elimination probably occurred by gene conversion. Also independently, the gene of Canis has added cysteine codons to the segment of repeats but has not yet lost cysteine 75. It is proposed that the gain and the loss of cysteine codons are parts of a multi-stage program of cysteine relocation.

Animals↗
PubMed1997

Blood metabolite and catecholamine responses to prolonged exercise following either acute plasma volume expansion or short-term training.

To determine the effect of acute plasma volume (PV) expansion on substrate utilization, blood metabolites and catecholamines to prolonged, moderate intensity cycle exercise, eight untrained men mean maximal oxygen uptake VO2max 4.10 (SEM 0.32) 1.min-1 were infused (10 ml.kg-1) with a 6% dextran (DEX) solution. These responses were also compared to those elicited using a short-term training (TR) protocol involving cycling for 90 to 120 min.day-1 at 60% VO2max for 3 consecutive days. In general DEX, which resulted in a calculated expansion of PV by 23.9% was without effect in modifying exercise oxygen uptake or the reduction in the respiratory exchange ratio (R) observed during prolonged exercise. In addition, the concentrations of blood glucose, glycerol, alanine and serum free fatty acids, although altered (P < 0.05) by exercise, were not altered by DEX. Blood lactate concentration was only higher (P < 0.05) at 30 min of exercise during DEX compared to the control. With the exception of blood lactate concentration, which was reduced (P < 0.05), TR did not change R or the concentrations of other blood metabolites. The concentrations of nonadrenaline and adrenaline, were depressed (P < 0.05) by DEX and TR at 60 and 90 min of exercise. These results would suggest that mechanisms as yet undefined can compensate for the estimated 10% reduction in arterial oxygen content mediated by acute PV expansion and enable prolonged exercise to be performed without adjustments in substrate selection and substrate mobilization.

Adult↗
PubMed1996

Peptides containing glutamine repeats as substrates for transglutaminase-catalyzed cross-linking: relevance to diseases of the nervous system.

Many proteins contain reiterated glutamine residues, but polyglutamine of excessive length may result in human disease by conferring new properties on the protein containing it. One established property of a glutamine residue, depending on the nature of the flanking residues, is its ability to act as an amine acceptor in a transglutaminase-catalyzed reaction and to make a glutamyl-lysine cross-link with a neighboring polypeptide. To learn whether glutamine repeats can act as amine acceptors, we have made peptides with variable lengths of polyglutamine flanked by the adjacent amino acid residues in the proteins associated with spinocerebellar ataxia type 1 (SCA1), Machado-Joseph disease (SCA3), or dentato-rubral pallidoluysian atrophy (DRPLA) or those residues adjacent to the preferred cross-linking site of involucrin, or solely by arginine residues. The polyglutamine was found to confer excellent substrate properties on any soluble peptide; under optimal conditions, virtually all the glutamine residues acted as amine acceptors in the reaction with glycine ethyl-ester, and lengthening the sequence of polyglutamine increased the reactivity of each glutamine residue. In the presence of transglutaminase, peptides containing polyglutamine formed insoluble aggregates with the proteins of brain extracts and these aggregates contained glutamyl-lysine cross-links. Repeated glutamine residues exposed on the surface of a neuronal protein should form cross-linked aggregates in the presence of any transglutaminase activated by the presence of Ca2+.

Amino Acid Sequence↗
PubMed1996

Comparative effects of acute volume expansion and short-term training on thermal and cardiovascular responses to prolonged exercise.

To investigate the hypothesis that increases in plasma volume (PV) are crucial to the cardiovascular and thermal adaptations resulting from training, eight moderately active males (Vo2max = 4.10 +/- 0.32 L/min; mean +/- SE) performed prolonged cycle exercise at 60% Vo2max during a control test (CON) and following infusion (10 mL/kg) of a 6% dextran solution (DEX). These responses were also compared with short-term training (TR) involving 3 days of cycling for 90-120 min at moderate intensity (60% Vo2max). During DEX, exercise cardiac output (Q) and stroke volume (SV) were persistently higher (p < 0.05) than CON, while heart rate (HR) was unchanged. In comparison, TR resulted in a lower (p < 0.05) HR at constant Q. In contrast with TR, in which the exercise response was unchanged from CON, the DEX condition produced a lower total peripheral resistance. Rectal temperature (Tre) was lowered (p < 0.05) both by DEX and TR, but the conditions differed in the time at which the reduction occurred. For DEX, the lower Tre was manifested early and persisted throughout exercise, whereas for TR the Tre was only lower later in exercise. Forearm blood flow, mean skin temperature, and sweat rate were not affected by DEX or TR. It is concluded that acute PV expansion does not affect the time-dependent response in cardiovascular function or thermoregulation during prolonged exercise in ambient conditions. The primary effects appear to be manifested during rest or soon after the onset of exercise.

Adult↗