Search PubMed⌕ Search

Biomedical subjects

H Lin

Publications and source records attributed to H Lin.

At least 37 records · Page 2Linked to original sources

Adhesive events in retinal development and function: the role of integrin receptors.

Cells in the developing retina contact a vast array of molecular cues in their microenvironment that are thought to guide their development. Many of these cues are embedded in the surface of neighboring cells or deposited within the extracellular matrix (ECM). Evidence has accumulated that cell-cell and cell-ECM interactions are essential in many phases of neural development, including neuroblast migration, determination of cell fate, axon outgrowth and synapse formation. In this chapter, we examine the developmental and functional roles fulfilled by integrins, a family of receptors for ECM molecules and cell adhesion molecules (CAMs). We have approached this problem by addressing a series of three questions: (1) which integrins are expressed in developing retina? (2) when and where are they expressed? and, (3) what functions do they carry out? Integrins have previously been implicated in axon extension, but new evidence suggests that they are also involved in earlier developmental events in preaxonal neuroblasts. High levels of expression of at least eight integrin subunits have been documented in these young retinal cells, and integrins containing the beta 1 subunit have been implicated in migration of adolescent retinal ganglion cells. Integrin expression persists through adulthood, both in the retina and in the neighboring layer of the retinal pigment epithelium (RPE). The integrin alpha v beta 5 has been shown to reside on the apical surface of the RPE and has been implicated in the phagocytosis of shed photoreceptor outer segments.

Animals↗

The role of radical surgery followed by adjuvant therapy for high-risk early-stage cervical carcinoma patients with pelvic lymph node metastasis.

OBJECTIVES: To identify a subgroup of high-risk node-positive patients in early-stage cervical cancers and to determine the role of radical hysterectomy followed by adjuvant therapy to these patients. STUDY DESIGN: We conducted a retrospective review of 482 surgically-treated patients of clinical stage Ib and IIa cervical carcinoma from July 1986 to December 1994 at Kaohsiung Chang Gung Memorial Hospital. Of these, 96 patients had pelvic lymph node metastases. Clinicopathological variables, including the level of pretreatment squamous cell carcinoma antigen (SCC-Ag), DNA flow cytometry analysis, and the use of different adjuvant therapies were studied. RESULTS: Disease-free survival was significantly worse among patients with S-phase fraction greater than 20% and pretreatment SCC-Ag level above 5 ng/ml. Utilizing these significant variables, we identified two distinct risk groups. Those patients without any of the risk variables were categorized as the low-risk group. Those patients with either one or both risk variables were categorized as the high-risk group. Five-year disease-free survival rates were 74% in the low-risk group and 43% in the high-risk group, (P=0.034). Disease recurred in 30.2% of the low-risk patients and 45. 3% of the high-risk patients. No survival advantages were found by using different adjuvant therapies. CONCLUSIONS: Radical hysterectomy should not be attempted if either the pretreatment SCC-Ag level is above 5 ng/ml or S-phase fraction of the tumor greater than 20% due to its limited value despite applying aggressive postoperative adjuvant therapy.

Adenocarcinoma↗

High potassium intake inhibits neointima formation in the rat carotid artery balloon injury model.

Recently, we reported that elevated extracellular potassium concentration in vitro inhibited proliferation and migration of vascular smooth muscle cells, formation of free radical compounds by macrophages, and reduced platelet sensitivity to agonists. In the present study we analyzed the effects of long-term, in vivo elevation of extracellular potassium concentration resulting from changes in dietary potassium intake on the vascular response to injury. The rat carotid artery balloon injury model was employed in 70 adult Sprague Dawley rats assigned to three groups. Beginning 14 days before surgical placement of the carotid lesion and continuing until death, the animals were fed diets containing either low (0.1% potassium, n = 25), normal (1.5% potassium, n = 19), or high potassium (4.0% potassium, n = 26). Fourteen days postsurgery the animals were killed and the arteries were analyzed to determine quantitatively the ratio of neointimal to medial area. Dietary potassium had a significant effect on arterial plasma potassium concentration (one-way analysis of variance, P < .01). Group mean and standard errors were 4.26+/-0.12 mmol/L for the low-potassium group, 5.22+/-0.19 mmol/L for normal, and 5.80+/-0.23 mmol/L for the high-intake group. Increases in dietary potassium attenuated neointima formation significantly (P < .05, one-way analysis of variance), with the mean ratio of neointimal area to medial area being 0.447+/-0.106 for the low-intake animals, 0.384+/-.116 for normal, and 0.240+/-.046 for the high-intake group. These results are consistent with a hypothesis that a high level of potassium intake is effective in inhibiting neointima formation in vivo.

Animals↗

The role of pretreatment squamous cell carcinoma antigen in predicting nodal metastasis in early stage cervical cancer.

PURPOSE: To evaluate whether the presence of pelvic lymph node metastasis can be predicted by pretreatment squamous cell carcinoma antigen (SCC-Ag) levels in early stage squamous cervical carcinoma. MATERIALS AND METHODS: Between 1994 and 1998, 284 patients with stage Ib and IIa cervical squamous cell carcinoma undergoing radical hysterectomy had preoperative SCC-Ag determination. The correlation between clinicopathological findings on SCC-Ag levels were examined. The Mann-Whitney U test was used to statistically analyze differences between node positive and negative patients. Multiple regression analysis and a multiple logistic model were employed to examine the effect of clinicopathological findings on SCC-Ag levels. RESULTS: Of the 284 patients, 56 patients were found to have nodal metastasis. Median serum levels and 90% ranges of SCC-Ag were 0.74 microg/l (0.5-7.8) in the 228 nodal negative patients and 4.33 microg/l (0.5-48.5) in the 56 nodal positive patients (p<0.001). Lymph node metastasis and tumor size were found to have a significant impact on SCC-Ag levels. Around 86% of the patients with SCC-Ag levels below 8 microg/l showed no nodal metastasis, while about 65% of the patients with serum levels above 8 microg/l exhibited nodal metastasis. Multivariate analyses confirmed that only lymph node metastasis had a significant impact on the SCC-Ag levels exceeding 8 microg/l. CONCLUSION: For predicting nodal metastasis preoperatively, SCC-Ag levels greater than 8 microg/ l can be considered a high-risk zone for nodal metastasis.

Antigens, Neoplasm↗

A syndrome of tricuspid atresia in mice with a targeted mutation of the gene encoding Fog-2.

Tricuspid atresia (TA) is a common form of congenital heart disease, accounting for 1-3% of congenital cardiac disorders. TA is characterized by the congenital agenesis of the tricuspid valve connecting the right atrium to the right ventricle and both an atrial septal defect (ASD) and a ventricular septal defect (VSD). Some patients also have pulmonic stenosis, persistence of a left-sided superior vena cava or transposition of the great arteries. Most cases of TA are sporadic, but familial occurrences with disease in multiple siblings have been reported. Gata4 is a zinc-finger transcription factor with a role in early cardiac development. Gata4-deficient mice fail to form a ventral heart tube and die of circulatory failure at embryonic day (E) 8.5 (refs 6,7). Zfpm2 (also known as Fog-2) is a multi-zinc-finger protein that is co-expressed with Gata4 in the developing heart beginning at E8.5 (refs 8-10). Zfpm2 interacts specifically with the N-terminal zinc finger of Gata4 and represses Gata4-dependent transcription. Here we use targeted mutagenesis to explore the role of Zfpm2 in normal cardiac development. Zfpm2-deficient mice died of congestive heart failure at E13 with a syndrome of tricuspid atresia that includes an absent tricuspid valve, a large ASD, a VSD, an elongated left ventricular outflow tract, rightward displacement of the aortic valve and pulmonic stenosis. These mice also display hypoplasia of the compact zone of the left ventricle. Our findings indicate the importance of Zfpm2 in the normal looping and septation of the heart and suggest a genetic basis for the syndrome of tricuspid atresia.

Animals↗

Direct effects of propylthiouracil on testosterone secretion in rat testicular interstitial cells.

The aim of this study was to investigate the mechanism by which propylthiouracil (PTU) exerts its inhibitory effects on the production of testosterone by rat testicular interstitial cells. The plasma testosterone concentration was decreased 60 and 120 min after an intravenous infusion of PTU (10 or 20 mg kg(-1)), but the concentration of plasma T(4) was unaffected by the drug treatment. Exposure of anterior pituitary tissue to PTU (3-12 mM) in vitro did not affect either basal or gonadotropin-releasing hormone (GnRH)-stimulated luteinizing hormone (LH) release. PTU (3 - 12 mM) inhibited both the basal and the human chorionic gonadotropin (hCG, 0.05 i.u. ml(-1))-stimulated release of testosterone from rat testicular tissue in vitro; at the highest concentration tested (12 mM), it also inhibited the forskolin or 8-bromo-adenosine 3':5'-cyclic monophosphate (8-Br-cyclic AMP)-stimulated release of testosterone. The 25-OH-cholesterol (10(-7)-10(-5) M)-stimulated release of pregnenolone and testosterone by the testicular interstitial cells was inhibited by PTU (12 mM, P<0.05). The results suggest that the inhibitory actions of PTU on testosterone secretion are exerted, at least in part, at the testicular level through a mechanism which is independent of thyroid status and which involves a reduction in P450scc activity and, hence, in the conversion of cholesterol to pregnenolone.

Animals↗

Identification of heading date quantitative trait locus Hd6 and characterization of its epistatic interactions with Hd2 in rice using advanced backcross progeny.

A backcrossed population (BC(4)F(2)) derived from a cross between a japonica rice variety, Nipponbare, as the recurrent parent and an indica rice variety, Kasalath, as the donor parent showed a long-range variation in days to heading. Quantitative trait loci (QTL) analysis revealed that two QTL, one on chromosome 3, designated Hd6, and another on chromosome 2, designated Hd7, were involved in this variation; and Hd6 was precisely mapped as a single Mendelian factor by using progeny testing (BC(4)F(3)). The nearly isogenic line with QTL (QTL-NIL) that carries the chromosomal segment from Kasalath for the Hd6 region in Nipponbare's genetic background was developed by marker-assisted selection. In a day-length treatment test, the QTL-NIL for Hd6 prominently increased days to heading under a 13.5-hr day length compared with the recurrent parent, Nipponbare, suggesting that Hd6 controls photoperiod sensitivity. QTL analysis of the F(2) population derived from a cross between the QTL-NILs revealed existence of an epistatic interaction between Hd2, which is one of the photoperiod sensitivity genes detected in a previous analysis, and Hd6. The day-length treatment tests of these QTL-NILs, including the line introgressing both Hd2 and Hd6, also indicated an epistatic interaction for photoperiod sensitivity between them.

Chromosome Mapping↗

Fresh and globular amyloid beta protein (1-42) induces rapid cellular degeneration: evidence for AbetaP channel-mediated cellular toxicity.

Amyloid beta peptides (AbetaP) deposit as plaques in vascular and parenchymal areas of Alzheimer's disease (AD) tissues and Down's syndrome patients. Although neuronal toxicity is a feature of late stages of AD, vascular pathology appears to be a feature of all stages of AD. Globular and nonfibrillar AbetaPs are continuously released during normal cellular metabolism, form calcium-permeable channels, and alter cellular calcium level. We used atomic force microscopy, laser confocal microscopy, and calcium imaging to examine the real-time and acute effects of fresh and globular AbetaP(1-42), AbetaP(1-40), and AbetaP(25-35) on cultured endothelial cells. AbetaPs induced morphological changes that were observed within minutes after AbetaP treatment and led to eventual cellular degeneration. Cellular morphological changes were most sensitive to AbetaP(1-42). AbetaP(1-42)-induced morphological changes were observed at nanomolar concentrations and were accompanied by an elevated cellular calcium level. Morphological changes were prevented by anti-AbetaP antibody, AbetaP-channel antagonist zinc, and the removal of extracellular calcium, but not by tachykinin neuropeptide, voltage-sensitive calcium channel blocker cadmium, or antioxidants DTT and Trolox. Thus, nanomolar fresh and globular AbetaP(1-42) induces rapid cellular degeneration by elevating intracellular calcium, most likely via calcium-permeable AbetaP channels and not by its interaction with membrane receptors or by activating oxidative pathways. Such rapid degeneration also suggests that the plaques, and especially fibrillar AbetaPs, may not have a direct causative role in AD pathogenic cascades.

Alzheimer Disease↗

Fresh and nonfibrillar amyloid beta protein(1-40) induces rapid cellular degeneration in aged human fibroblasts: evidence for AbetaP-channel-mediated cellular toxicity.

Alzheimer's disease (AD) is primarily nonfamilial or sporadic (SAD) in origin, although several genetic linkages are reported. Tissues from AD patients contain fibrillar plaques made of 39 to 43 amino acid-long amyloid beta peptide (AbetaP), although the mechanisms of AbetaP toxicity are poorly understood. AbetaP(1-40) is the most prevalent AbetaP present in the neuronal and non-neuronal tissues from SAD patients. AbetaP(1-40) toxicity has been examined mainly after prolonged incubation and correlates with the age and fibrillar morphology of AbetaP(1-40). Globular and nonfibrillar AbetaPs are released continually during normal cellular metabolism; they elevate cellular Ca(2+) and form cation-permeable channels. However, their role in cellular toxicity is poorly understood. We have used an integrated atomic force and light fluorescence microscopy (AFM-LFM), laser confocal microscopy, and calcium imaging to examine real-time and acute effect of fresh and globular AbetaP(1-40) on cultured, aged human, AD-free fibroblasts. AFM images show that freshly prepared AbetaP(1-40) in phosphate-buffered saline (PBS) are globular and do not form fiber for an extended time period. AbetaP(1-40) induced rapid structural modifications, including cytoskeletal reorganization, retraction of cellular processes, and loss of cell-cell contacts, within minutes of incubation. This led to eventual cellular degeneration. AbetaP(1-40)-induced degeneration was prevented by anti-AbetaP antibody, zinc, and Tris, but not by tachykinin neuropeptides. In Ca(2+)-free extracellular medium, AbetaP(1-40) did not induce cellular degeneration. In the presence of extracellular Ca(2+), AbetaP(1-40) induced a sustained increase in the cellular Ca(2+). Thus, short-term and acute AbetaP(1-40) toxicity is mediated by Ca(2+) uptake, most likely via calcium-permeable AbetaP pores. Such rapid degeneration does not require fibrillar plaques, suggesting that the plaques may not have any causative role.

Alzheimer Disease↗

Cell-penetrating SH3 domain blocker peptides inhibit proliferation of primary blast cells from CML patients.

Bcr-Abl contributes prominently to the development of most chronic myeloid leukemias (CMLs). Prior work has identified the adapter protein CRKL as a major substrate of the Bcr-Abl tyrosine kinase. CRKL can also bind via its first SH3 domain [SH3(1)] to specific sequences in Bcr-Abl. Cell-penetrating peptides were developed that bind with high affinity and selectivity to the SH3(1) domain of CRKL. They disrupt Bcr-Abl-CRKL complexes and strongly reduce the proliferation of primary CML blast cells and cell lines established from Bcr-Abl-positive patients. Activation-specific antibodies against phosphorylated MAP kinase (MAPK) showed that MAPK activity is down-regulated in blast cells treated with the CRKLSH3(1) blocker peptides. We conclude that the Bcr-Abl-CRKL complexes are largely dependent on the CRKLSH3(1) domain, that the central mitogenic cascade is down-regulated as a consequence of the disruption of CRKLSH3(1) interactions, and that CRKL therefore contributes to the proliferation of CML blast cells.

Adaptor Proteins, Signal Transducing↗

Dietary Bifidobacterium lactis (HN019) enhances resistance to oral Salmonella typhimurium infection in mice.

The ability of a newly identified probiotic lactic acid bacterial strain, Bifidobacterium lactis (HN019), to confer protection against Salmonella typhimurium was investigated in BALB/c mice. Feeding mice with B. lactis conferred a significant degree of protection against single or multiple oral challenge with virulent S. typhimurium, in comparison to control mice that did not receive B. lactis. Protection included a ten-fold increase in survival rate, significantly higher post-challenge food intake and weight gain, and reduced pathogen translocation to visceral tissues (spleen and liver). Furthermore, the degree of pathogen translocation showed a significant inverse correlation with splenic lymphocyte proliferative responses to mitogens, blood and peritoneal cell phagocytic activity and intestinal mucosal anti-S. typhimurium antibody titers in infected mice; all of these immune parameters were enhanced in mice fed B. lactis. Together, these results suggest that dietary B. lactis can provide a significant degree of protection against Salmonella infection by enhancing various parameters of immune function that are relevant to the immunological control of salmonellosis. Thus dietary supplementation with B. lactis provides a unique opportunity for developing immune-enhancing probiotic dairy food products with proven health benefits.

Animals↗

Phospholipase C is involved in kinetochore function in Saccharomyces cerevisiae.

The budding yeast PLC1 gene encodes a homolog of the delta isoform of mammalian phosphoinositide-specific phospholipase C. Here, we present evidence that Plc1p associates with the kinetochore complex CBF3. This association is mediated through interactions with two established kinetochore proteins, Ndc10p and Cep3p. We show by chromatin immunoprecipitation experiments that Plc1p resides at centromeric loci in vivo. Deletion of PLC1, as well as plc1 mutations which abrogate the interaction of Plc1p with the CBF3 complex, results in a higher frequency of minichromosome loss, nocodazole sensitivity, and mitotic delay. Overexpression of Ndc10p suppresses the nocodazole sensitivity of plc1 mutants, implying that the association of Plc1p with CBF3 is important for optimal kinetochore function. Chromatin extracts from plc1Delta cells exhibit reduced microtubule binding to minichromosomes. These results suggest that Plc1p associates with kinetochores and regulates some aspect of kinetochore function and demonstrate an intranuclear function of phospholipase C in eukaryotic cells.

Centromere↗

Divergent N-terminal sequences target an inducible testis deubiquitinating enzyme to distinct subcellular structures.

Ubiquitin-specific processing proteases (UBPs) presently form the largest enzyme family in the ubiquitin system, characterized by a core region containing conserved motifs surrounded by divergent sequences, most commonly at the N-terminal end. The functions of these divergent sequences remain unclear. We identified two isoforms of a novel testis-specific UBP, UBP-t1 and UBP-t2, which contain identical core regions but distinct N termini, thereby permitting dissection of the functions of these two regions. Both isoforms were germ cell specific and developmentally regulated. Immunocytochemistry revealed that UBP-t1 was induced in step 16 to 19 spermatids while UBP-t2 was expressed in step 18 to 19 spermatids. Immunoelectron microscopy showed that UBP-t1 was found in the nucleus while UBP-t2 was extranuclear and was found in residual bodies. For the first time, we show that the differential subcellular localization was due to the distinct N-terminal sequences. When transfected into COS-7 cells, the core region was expressed throughout the cell but the UBP-t1 and UBP-t2 isoforms were concentrated in the nucleus and the perinuclear region, respectively. Fusions of each N-terminal end with green fluorescent protein yielded the same subcellular localization as the native proteins, indicating that the N-terminal ends were sufficient for determining differential localization. Interestingly, UBP-t2 colocalized with anti-gamma-tubulin immunoreactivity, indicating that like several other components of the ubiquitin system, a deubiquitinating enzyme is associated with the centrosome. Regulated expression and alternative N termini can confer specificity of UBP function by restricting its temporal and spatial loci of action.

Age Factors↗

piwi encodes a nucleoplasmic factor whose activity modulates the number and division rate of germline stem cells.

piwi represents the first class of genes known to be required for stem cell self-renewal in diverse organisms. In the Drosophila ovary, piwi is required in somatic signaling cells to maintain germline stem cells. Here we show that piwi encodes a novel nucleoplasmic protein present in both somatic and germline cells, with the highly conserved C-terminal region essential for its function. Removing PIWI protein from single germline stem cells significantly decreases the rate of their division. This suggests that PIWI has a second role as a cell-autonomous promoter of germline stem cell division. Consistent with its dual function, over-expression of piwi in somatic cells causes an increase both in the number of germline stem cells and the rate of their division. Thus, PIWI is a key regulator of stem cell division - its somatic expression modulates the number of germline stem cells and the rate of their division, while its germline expression also contributes to promoting stem cell division in a cell-autonomous manner.

Animals↗

[Study of covalently immobilizing heparin to polyetherurethane surface].

In this paper an approach of covalently immobilizing heparin to the surface of polyetherurethane without functional groups was studied. By quantitation of amino groups on PEU surface, the appropriate hydrolysis condition to PEU was determined. The assays of the heparin content heparinized PEU and the time of recalcification proved the above approach comparatively ideal.

Heparin↗

A magnetic field-responsive domain in the human HSP70 promoter.

HSP70 gene expression is induced by a wide range of environmental stimuli, including 60-Hz electromagnetic fields. In an earlier report we showed that the induction of HSP70 gene expression by magnetic fields is effected at the level of transcription and is mediated through c-myc protein binding at two nCTCTn sequences at -230 and -160. in the human HSP70 promoter. We report on the identification of a third c-myc binding site (between -158 and -162) that is an important regulator of magnetic field-induced HSP70 expression. We also show that the heat shock element (HSE), lying between -180 and -203, is required for induction of HSP70 gene expression by magnetic fields. The HSE centered at -100 alone is insufficient.

Binding Sites↗

Amyloid beta protein (1-40) forms calcium-permeable, Zn2+-sensitive channel in reconstituted lipid vesicles.

Amyloid beta protein (A beta P) forms senile plaques in the cerebrocortical blood vessels and brain parenchyma of patients with Alzheimer's disease (AD). The nonfamilial or sporadic AD (SAD), the most prevalent form of AD, has been correlated with an increased level of 40-residue A beta P (A beta P1-40). However, very little is known about the role of A beta P1-40 in AD pathophysiology. We have examined the activity of A beta P1-40 reconstituted in phospholipid vesicles. A combined light fluorescence and atomic force microscope (AFM) was used to image the structure of reconstituted vesicles and 45Ca2+ uptake was used as an assay for calcium permeability across the vesicular membrane. Vesicles reconstituted with fresh and globular A beta P1-40 contain a significant amount of A0 beta P and exhibit strong immunofluorescence labeling with an antibody raised against the N-terminal domain of A beta P, suggesting the incorporation of A beta P1-40 peptide in the vesicular membrane. Vesicles reconstituted with A beta P1-40 exhibited a significant level of 45Ca2+ uptake. The vesicular calcium level saturated over time, showing an important ion channel characteristic. The 45Ca2+ uptake was inhibited by (i) a monoclonal antibody raised against the N-terminal region of A beta P and (ii) Zn2+. However, a reducing agent (DTT) did not inhibit the 45Ca2+ uptake, indicating that the oxidation of A beta P or its surrounding lipid molecules is not directly involved in A beta P-mediated Ca2+ uptake. These findings provide biochemical and structural evidence that fresh and globular A beta P1-40 forms calcium-permeable channels and thus may induce cellular toxicity by regulating the calcium homeostasis in nonfamilial or sporadic Alzheimer's disease.

Amyloid beta-Peptides↗