Biomedical subjects

K Guo

Publications and source records attributed to K Guo.

At least 37 records · Page 2Linked to original sources

PubMed1999

The membrane association domain of RGS16 contains unique amphipathic features that are conserved in RGS4 and RGS5.

Regulators of G protein signaling (RGS proteins) modulate G protein-mediated signaling pathways by acting as GTPase-activating proteins for Gi, Gq, and G12 alpha-subunits of heterotrimeric G proteins. Although it is known that membrane association is critical for the biological activities of many RGS proteins, the mechanism underlying this requirement remains unclear. We reported recently that the NH2 terminus of RGS16 is required for its function in vivo. In this study, we show that RGS16 lacking the NH2 terminus is no longer localized to the plasma membrane as is the wild type protein, suggesting that membrane association is important for biological function. The region of amino acids 7-32 is sufficient to confer the membrane-targeting activity, of which amino acids 12-30 are predicted to adopt an amphipathic alpha-helix. Site-directed mutagenesis experiments showed that the hydrophobic residues of the nonpolar face of the helix and the strips of positively charged side chains positioned along the polar/nonpolar interface of the helix are crucial for membrane association. Subcellular fractionation by differential centrifugation followed by conditions that distinguish peripheral membrane proteins from integral ones indicate that RGS16 is a peripheral membrane protein. We show further that RGS16 membrane association does not require palmitoylation. Our results, together with other recent findings, have defined a unique membrane association domain with amphipathic features. We believe that these structural features and the mechanism of membrane association of RGS16 are likely to apply to the homologous domains in RGS4 and RGS5.

Amino Acid Sequence↗

PubMed1999

Isolation of spermidine synthase gene (spsA) of Dictyostelium discoideum.

The gene encoding spermidine synthase (spsA) was isolated from Dictyostelium discoideum using the technique of insertional mutagenesis. Northern blot analysis showed that the spsA mRNA is expressed maximally during the vegetative stage and decreases gradually during the 24 h of development. Sequencing of the genomic DNA and a full-length cDNA clone indicated the presence of one intron in a gene coding for a predicted protein (SpsA) with 284 amino acids. The sequence is highly conserved, with amino acid identities compared to spermidine synthases of humans, 59.5%, to mouse, 61.3%, and to yeast, 58.1%. A null mutant of the spsA gene is unable to grow in the absence of exogenous spermidine. Development of spsA null cells grown in the absence of spermidine produced fruiting bodies that have abnormally short stalks.

Amino Acid Sequence↗

PubMed1999

Grating and plaid chrominance motion influences the suppressed ocular following response.

Involuntary eye movements to foveal stimulation were measured in a monkey while it performed a fixation task. Second-order plaid motion generated higher velocities of eye movements than did first-order gratings, yet the latency of the early following response was no different for grating or plaid motion. Nevertheless, early suppressed ocular following responses to isoluminant motion continue to be titrated by stimulus velocity and spatial frequency. Motion defined by 60% luminance contrast gratings and plaids generated a motion signal gain of 60% over chrominance motion. The 20% longer latency of eye movements to chrominance motion may reflect the longer conduction latency of the parvocellular channel and an additional stage in cortical processing en route to motion areas and eye movement control.

Analysis of Variance↗

PubMed1999

Cell cycle withdrawal promotes myogenic induction of Akt, a positive modulator of myocyte survival.

During myogenesis, proliferating myoblasts withdraw from the cell cycle, acquire an apoptosis-resistant phenotype, and differentiate into myotubes. Previous studies indicate that myogenic induction of the cyclin-dependent kinase inhibitor p21 results in an inhibition of apoptotic cell death in addition to its role as a negative cell cycle regulator. Here we demonstrate that the protein encoded by the Akt proto-oncogene is induced in C2C12 cells during myogenic differentiation with a corresponding increase in kinase activity. In differentiating cultures, expression of dominant-negative forms of Akt increase the frequency of cell death whereas expression of wild-type Akt protects against death, indicating that Akt is a positive modulator of myocyte survival. Antisense oligonucleotides against p21 block cell cycle withdrawal, inhibit Akt induction, and enhance cell death in differentiating myocyte cultures. Adenovirus-mediated transfer of wild-type or constitutively active Akt constructs confer partial resistance to cell death under conditions where cell cycle exit is blocked by the antisense oligonucleotides. Collectively, these data indicate that cell cycle withdrawal facilitates the induction of Akt during myogenesis, promoting myocyte survival.

Adenoviruses, Human↗

PubMed1999

Vascular smooth muscle cell growth arrest on blockade of thrombospondin-1 requires p21(Cip1/WAF1).

Abnormal proliferation of vascular smooth muscle cells (VSMCs) is thought to play an important role in the pathogenesis of atherosclerosis and restenosis. Previous studies have implicated the extracellular matrix protein thrombospondin-1 (TSP1) in mitogen-dependent proliferation of VSMCs. In this study, we investigated the molecular mechanisms involved in TSP1-mediated regulation of VSMC growth. Neutralizing A4.1 anti-TSP1 antibody inhibited the activity of the G(1)/S cyclin-dependent kinase 2 (cdk2) and blocked the induction of S-phase entry, which normally occurs in serum-stimulated VSMCs. This growth-inhibitory effect was associated with a marked induction of p21(Cip1/WAF1) (p21) expression in A4.1-treated VSMCs. Moreover, addition of A4.1 antibody to VSMCs markedly increased the level of p21 bound to cdk2. Thus growth arrest on antibody blockade of TSP1 may be mediated by the cdk inhibitory protein p21. Consistent with this notion, anti-TSP1 antibody inhibited [(3)H]-thymidine incorporation in wild-type but not in p21-deficient mouse embryonic fibroblasts (MEFs). Together, these data suggest that p21 plays an important role in TSP1-mediated control of cellular proliferation.

Animals↗

PubMed1999

A myb-related protein required for culmination in Dictyostelium.

The avian retroviral v-myb gene and its cellular homologues throughout the animal and plant kingdoms contain a conserved DNA binding domain. We have isolated an insertional mutant of Dictyostelium unable to switch from slug migration to fruiting body formation i.e. unable to culminate. The gene that is disrupted, mybC, codes for a protein with a myb-like domain that is recognized by an antibody against the v-myb repeat domain. During development of myb+ cells, mybC is expressed only in prestalk cells. When developed together with wild-type cells mybC- cells are able to form both spores and stalk cells very efficiently. Their developmental defect is also bypassed by overexpressing cAMP-dependent protein kinase. However even when their defect is bypassed, mybC null slugs and culminates produce little if any of the intercellular signalling peptides SDF-1 and SDF-2 that are believed to be released by prestalk cells at culmination. We propose that the mybC gene product is required for an intercellular signaling process controlling maturation of stalk cells and spores and that SDF-1 and/or SDF-2 may be implicated in this process.

Amino Acid Sequence↗

PubMed1999

U.S. academic medical centers under the managed health care environment.

This research investigates the impact of managed health care on academic medical centers in the United States. Academic medical centers hold a unique position in the U.S. health care system through their missions of conducting cutting-edge biomedical research, pursuing clinical and technological innovations, providing state-of-the-art medical care and producing highly qualified health professionals. However, policies to control costs through the use of managed care and limiting resources are detrimental to academic medical centers and impede the advancement of medical science. To survive the threats of managed care in the health care environment, academic medical centers must rely on their upper level managers to derive successful strategies. The methods used in this study include qualitative approaches in the form of key informants and case studies. In addition, a survey questionnaire was sent to 108 CEOs in all the academic medical centers in the U.S. The findings revealed that managers who perform the liaison, monitor, entrepreneur and resource allocator roles are crucial to ensure the survival of academic medical centers, so that academic medical centers can continue their missions to serve the general public and promote their well-being.

Academic Medical Centers↗

PubMed1998

Short-latency ocular following of motion by monkeys during a fixation task.

Short-latency ocular following responses in monkeys, induced by first- and second-order motion during a fixation task, were sensitive to stimulus size, retinal eccentricity and motion duration. Latency of eye movements was no different for first- or second-order motion and simply marked the occurrence of movement in the visual field. Second-order motion produced higher velocity eye movements than did first-order motion. The velocity of eye movements was influenced by the orientation of second-order pattern components as well as their conspicuous energy. Even when partially suppressed by a fixation task, involuntary eye movements during the initial pursuit remain sensitive to several properties of visual motion.

Analysis of Variance↗

PubMed1998

Involuntary eye movements in response to first- and second-order motion.

We trained monkeys to maintain fixation while first- and second-order motion stimuli were displayed centrally in the visual field. Stimulus velocity, spatial frequency and contrast were varied to determine differences in patterns of involuntary eye movements elicited by random onset of stimulus motion. We observed different patterns of eye movement latency and velocity suggesting very early (< or = 100 ms) components of oculomotor activity are used to initiate smooth pursuit of object trajectory. Eye movement latency was insensitive to the complexity of stimulus motion, whereas second-order motion elicited faster eye movements than first-order motion. Instantaneous eye movement velocity might be related to the earliest stages of visual processing of component motion.

Animals↗

PubMed1998

Residual motion discrimination using colour information without primary visual cortex.

Previous studies have reported that some patients with damage to striate cortex retain the ability to detect monochromatic light and discriminate direction of achromatic movement in their blind visual fields. We investigated the residual chromatic visual capacity of a well-studied patient (GY) who has a unilateral lesion to striate cortex (V1). The data demonstrated that GY was able to detect and discriminate isoluminant colour targets presented in his blind hemifield. The velocity and contrast of chromatic moving stimuli affected GY's levels of conscious experience of movements he was able to discriminate. As the velocity or contrast was increased, his discrimination performance improved in line with rising visual awareness and judgement confidence. At isoluminance, GY continued to discriminate motion direction with above chance accuracy. These results indicate chromatic signals can also be used to process motion information in the absence of primary visual cortex.

Adult↗

PubMed1998

Nitric oxide-induced downregulation of Cdk2 activity and cyclin A gene transcription in vascular smooth muscle cells.

BACKGROUND: Nitric oxide (NO) inhibits vascular smooth muscle cell (VSMC) proliferation and neointima formation after balloon injury. However, the molecular mechanisms underlying NO-mediated growth arrest are poorly understood. In the present study, we examined the effects of the NO donors sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP) on cell cycle activity in VSMCs. METHODS AND RESULTS: Stimulation of quiescent rat VSMCs with serum leads to an increase in cyclin-dependent kinase (cdk)2 kinase activity that correlates with a marked induction of cyclin A protein expression. The addition of SNP or SNAP to VSMC cultures at the time of serum stimulation abrogates the induction of cdk2 activity without suppressing protein levels of cdk2 or cyclin E. These NO donors block serum-stimulated upregulation of cyclin A mRNA and protein and repress the serum induction of cyclin A promoter activity in VSMCs. CONCLUSIONS: The addition of the nitric oxide donors SNP or SNAP to mitogen-stimulated VSMCs prevents activation of cdk2, a key regulator of the G1 and S phases of the cell cycle. These NO donors do not affect the expression of cdk2 protein but block the mitogen-induced expression of cyclin A, an activating subunit of cdk2. SNP and SNAP also repress the mitogen-stimulated activation of the cyclin A promoter. These data suggest that the antiproliferative effect of NO on VSMCs results, at least in part, from the repression of cyclin A gene transcription.

Animals↗

PubMed1998

Direction discrimination of moving gratings and plaids and coherence in dot displays without primary visual cortex (V1).

We present new experimental observations of G.Y., a well-tested patient with unilateral loss of primary visual cortex. We stimulated G.Y.'s blind hemifield using first- and second-order motion stimuli at velocities around psychophysical threshold. Using a dual response paradigm (awareness level of visual motion, motion direction discrimination) psychophysical performance improved with increasing velocity and dot coherence. We were also able to influence directly G.Y.'s performance for the better and at will, by placing the emphasis solely on direction discrimination. In the absence of V1, graduated detection and discrimination of stimuli known to activate both V1 and extrastriate motion areas MT/V5 and MST is still possible. These results are in line with residual visual processing but did not show evidence of unconscious processing of motion stimuli characteristic of 'blindsight'.

Adult↗

PubMed1998

Apoptosis-associated gene expression in the corpus luteum of the rat.

The involution of the corpus luteum (CL) at parturition is an example of physiological apoptosis, a complex process involving massive vascular regression while luteal cells undergo apoptosis. In the present study, changes in gene expression associated with physiological apoptosis were examined. Three genes isolated in our laboratory because of their association with apoptotic processes in the ovary, mammary gland, and prostate served as the focus of our investigation: Y81, Gas-1, and the gene IAP encoding integrin-associated protein. Y81 is a novel gene for which three transcripts are apparent. A Y81 cDNA clone representing the longest transcript has been isolated; it shows an open reading frame exhibiting a region of very high homology with members of the frizzled family, the prototypes of which are cell autonomous polarity genes encoding seven-pass transmembrane receptor proteins, for example the receptor for Wingless. Gas-1 is known as a growth-arrest gene that inhibits DNA synthesis when microinjected into cells. Integrin-associated protein is a beta 3-integrin-binding protein for which, recently, a thrombo-spondin-binding activity has been recognized. These three genes, all sharply up-regulated in the course of physiological involution processes in the ovarian CL, in mammary gland, and in prostate, seem promising candidates-by virtue of their specific expression in distinct tissues undergoing programmed cell death-as mediators of stimuli leading to apoptosis and subsequent phagocytosis. In this study, sulfated glycoprotein-2, previously observed in many instances of physiological apoptosis, was further employed as an indicator for incipient apoptosis, and stromelysin was followed as a marker for the tissue remodeling activity that is intimately associated with apoptosis during involution.

Amino Acid Sequence↗

PubMed1998

Cellular composition and anatomic distribution in nonfunctioning pancreatic endocrine tumors: immunohistochemical study of 30 cases.

OBJECTIVE: To investigate the cytological pattern and distribution in nonfunctioning pancreatic endocrine tumors. METHODS: Using labeled streptavidin-biotin (LSAB), immunohistochemical staining for insulin, glucagon, somatostatin, pancreatic polypeptide and gastrin was performed on 30 nonfunctioning pancreatic endocrine tumors from 30 patients. The cellular composition and anatomic distribution in these tumors were analyzed. RESULTS: Of 30 tumor tissues, 22 (73.3%) were found to contain cells immunoreactive to 1-4 kinds of peptide hormones; 17 (56.7%) showed positive staining for more than one peptide and up to 4 peptides; and 8 (26.7%) showed negative immunoreaction to all antiserum applied. No tumor was found to contain immunoreactive gastrin. Among 17 multihormonal tumors, 4 contained 2 kinds of peptide hormones, 8 had 3 kinds, and 5 harbored 4 kinds of peptide hormones. In addition, the difference in the number and type of positive endocrine cells between the tumors arising from the head of the pancreas and those arising from the body and tail of the pancreas were statistically significant (P < 0.05). CONCLUSIONS: Immunohistochemically, the high positive rate to peptide hormones suggests that the nonfunctioning pancreatic endocrine tumors are actually not nonfunctioning; they are asymptomatic pancreatic endocrine tumors. Moreover, an uneven distribution of positive endocrine cells in the nonfunctioning pancreas endocrine tumors within the pancreas was identified.

Adenoma, Islet Cell↗

PubMed1998

[The general survey for chondromalacia of 2,743 Chinese population].

OBJECTIVE: To evaluate the distribution of chondromalacia patella in Chinese population. METHODS: A random cluster sampling survey was performed covering 2,743 subjects varied in age, sex and occupation in 1995. RESULTS: The prevalence rate is 36.2%. The occurrence in women was higher than that in men (P < 0.01), while in the age group, 30 to 39 years was the highest being 55.8%. The prevalence rate in soldier being 47.5% was the highest among varied occupations. CONCLUSIONS: Our study is the first survey to be performed in a large number of Chinese population. This investigation may reflect the prevalence rate of chondromalacia patella in China.

Adolescent↗

PubMed1998

[PCR in detecting the correlation between infection of HBV and cholelithiasis].

OBJECTIVE: To study the correlation between cholelithiasis and the infection of HBV. METHOD: 32 formalin-fixed and paraffin-embedded gallbladder samples of cholelithiasis patients and 20 gallbladder samples of non-cholelithiasis patients were investigated using the polymerase chain reaction-Ethidium bromide (PCR-EB) assay. The 52 patients were positive to HBV serologic markers. RESULT: The results showed that HBV-DNA was found in 13 gallbladder samples of 32 cholelithiasis patients (40.63%), significantly higher than that in 3 gallbladder samples of 20 non-cholelithiasis patients (15%). CONCLUSION: The infection of HBV and the formation of cholelithiasis are correlated.

Adult↗

PubMed1997

p21CIP1-mediated inhibition of cell proliferation by overexpression of the gax homeodomain gene.

gax, a diverged homeobox gene expressed in vascular smooth muscle cells (VSMCs), is down-regulated in vitro by mitogen stimulation and in vivo in response to vascular injury that leads to cellular proliferation. Recombinant Gax protein microinjected into VSMCs and fibroblasts inhibited the mitogen-induced entry into S-phase when introduced either during quiescence or early stages of G1. Overexpression of gax with a replication-defective adenovirus vector resulted in G0/G1 cell cycle arrest of VSMCs and fibroblasts. The gax-induced growth inhibition correlated with a p53-independent up-regulation of the cyclin-dependent kinase inhibitor p21. Gax overexpression also led to an association of p21 with cdk2 complexes and a decrease in cdk2 activity. Fibroblasts deficient in p21 were not susceptible to a reduction in cdk2 activity or growth inhibition by gax overexpression. Localized delivery of the virus to denuded rat carotid arteries significantly reduced neointima formation and luminal narrowing. These data indicate that gax overexpression can inhibit cell proliferation in a p21-dependent manner and can modulate injury-induced changes in vessel wall morphology that result from excessive cellular proliferation.

Adenoviridae↗

PubMed1997

Eye position-dependent activation of neurones in striate cortex of macaque.

Extracellular recordings were made in the striate cortex in awake, behaving monkeys to test the influence of the eye position on cellular activity. Two monkeys (Macaca mulatta) were trained to fixate a small spot at 25 different eye positions. About half (52%) of the studied neurones showed a selective gaze field (GF). The cells' activities increased significantly when the monkey fixated at this field of view. For the majority of these neurones, GFs were located at the contralateral field of view with respect to the hemisphere from which responses were recorded, and were usually found a few degrees peripheral to the related receptive field. Eye position-dependent neurones were found at different depths of cortex, but mostly in the superficial layers. The results indicate that some neurones in striate cortex may code information about eye position and could contribute to target localization in a head-centred coordinate system by combining retinal and afferent eye position signals.

Analysis of Variance↗