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Biomedical subjects

H Tseng

Publications and source records attributed to H Tseng.

At least 19 recordsLinked to original sources
PubMed2001

Effects of surface treatments on bond strength of glass-infiltrated ceramic.

The purpose of this study was to evaluate the effects of various surface treatments on the bond strength at the In-Ceram/resin composite interface. Ninety-eight In-Ceram specimens were divided into seven groups and exposed to various surface treatments as follows: (A) control (B) saliva contamination (C) saliva contamination plus aluminum oxide sandblasting (D) glove powder contamination (E) glove powder contamination plus aluminum oxide sandblasting (F) rough aluminum oxide sandblasting and (G) excess glass infiltration. A resin composite cylinder was cemented to each In-Ceram specimen with Panavia 21 resin luting cement. Half of the cemented specimens in each group were stored in water for 24 h, and the other half were stored in water for 2 weeks and then were thermo-cycled for 2000 cycles. Shear bond strengths (SBS) of seven specimens in each subgroup were determined and analysed using analysis of variance (ANOVA) and Tukey HSD test as well as Student's t-test. Scanning electronic microscopy was used to identify the type of bond failure. Shear bond strength was significantly decreased by saliva and glove powder contaminations (P < 0.05). Sandblasting treatment did not improve the saliva-contaminated specimens. However, the glove powder plus sandblasting group showed no significant difference in SBS compared with the control group. There was no significant difference in SBS between the excess glass-infiltrating group and the control group. The SBS was significantly decreased by rough aluminum oxide sandblasting (P < 0.05). The SBS values of groups without thermocycling were significantly greater than those of groups with thermocycling (P < 0.05). There were no significant differences among SBS values of the seven groups with thermocycling. Combined cohesive and adhesive bond failures were seen in every group. Various surface treatments or contaminants may significantly influence the bond strength of In-Ceram restorative in clinical use.

Composite Resins↗
PubMed2001

Function of basonuclin in increasing transcription of the ribosomal RNA genes during mouse oogenesis.

Active protein synthesis during early oogenesis requires accelerated transcription of ribosomal RNA genes (rDNAs). In response to this demand, rDNAs are amplified more than 1000-fold early in Xenopus oogenesis. Here, we report evidence that rDNA is not amplified in mouse oocytes, but these cells may instead employ the zinc-finger protein basonuclin, a putative rDNA transcription factor, to enhance rRNA synthesis. This conclusion is based on observations that basonuclin is localized in the nucleolus in the mouse oocyte early in its growth phase, when rRNA transcription is highly active; and that the binding sites of basonuclin zinc fingers on the human and mouse rDNA promoters are homologous. In a co-transfection assay, basonuclin can elevate transcription from an rDNA promoter, and its zinc-finger domain can inhibit RNA polymerase I transcription, as detected by a run-on assay, in growing mouse oocytes.

Animals↗
PubMed2000

Effect of temperature and flux concentration on soldering of base metal.

The present study used the acoustic emission (AE) technique to evaluate interactions among soldering temperature, flux treatment, and the resultant ultimate tensile strength (UTS). Scanning electron microscopy (SEM) was used to examine fracture surfaces of the solder joints. Specimens were cast from removable partial denture alloy and then placed in a jig with a gap distance of 1.0 mm. A high-frequency soldering machine with an optical pyrometer was used for soldering at 1150 degrees C and 1200 degrees C, respectively. The flux concentrations were 67% and 75%. The soldered specimens were subjected to tensile test at a crosshead speed of 0.05 mm/min. During testing, acoustic emissions in the frequency range of 100--1200 kHz were collected, filtered, recorded, and processed by a sensing device. The results were analysed by ANOVA and Tukey LSD test. UTS at different temperatures showed no significant difference according to either mechanical or acoustic results. But in the 1200 degrees C group, the UTSs and AE counts showed significant differences (P<0.05) at both flux concentrations. SEM showed that the 1200C group had better dendritic crystal structure than did the 1150 degrees C group. In the 1200 degrees C group specimens with 67% flux had fewer flux inclusion bodies and dendritic crystals than did specimens with 75% flux. The 75% flux subgroup produced high-amplitude (60--70 dB) acoustic signals within the elastic deformation zone, while the 67% flux subgroup produced similar signals within the plastic deformation zone, either beyond the 0.2% yield point or before fracture.

Acoustics↗
PubMed1999

A GC-rich sequence within the 5' untranslated region of human basonuclin mRNA inhibits its translation.

By the method of RNase protection, the 5' ends of the basonuclin mRNA were mapped to four sites distributed over 400 bases of the genomic sequence, a result implying four different basonuclin transcripts within the cell. Despite the heterogeneity at the 5' end, all four basonuclin mRNA shared the same translation initiation codon. However, only two transcripts contained, in their 5' untranslated region (UTR), a GC-rich sequence of approx. 180 bases. The ability of this GC-rich sequence to form a large and stable secondary structure was suggested by experimental results from primer extension, RNase resistance, and computer analysis of the sequence. In vitro study showed that translation of basonuclin RNA containing this putative structure could not be initiated efficiently from the first two AUGs, whereas those that lacked it could.

5' Untranslated Regions↗
PubMed1999

Basonuclin in murine corneal and lens epithelia correlates with cellular maturation and proliferative ability.

Basonuclin is a zinc finger protein with highly restricted tissue distribution. It has been found in abundance only in keratinocytes of stratified epithelia and the germ cells of the testis and ovary. We studied the expression pattern of basonuclin in relation to cellular proliferation and differentiation in murine corneal and lens epithelia, two self-renewing tissues in the eye which contain cells that proliferate throughout life. Mouse corneal and lens epithelial cells at various stages of development were labeled with BrdU for 90 min to detect cells in S phase and to establish proliferative rates. Whole eyes of mouse or rat were processed for frozen sections and cellular basonuclin was detected by either a rabbit antimouse- or a rabbit anti-human-basonuclin antibody. Basonuclin was expressed in virtually all cells in the basal layer of corneal epithelium and in the pre-equatorial lens epithelium, the respective proliferative compartments of adult corneal and lens epithelia. Basonuclin expression in corneal epithelium began at post-natal life day 4, first in a few cells and then spread to virtually all basal cells at day 20. Basonuclin was consistently absent in limbal epithelium. Lens basonuclin, which was detected earlier than that of the cornea, was confined to the pre-equatorial epithelium and was absent in equatorial cells that expressed p57KIP2, an early differentiation marker for these cells. An important distinction between corneal and lens basonuclin is that the former is predominantly nuclear whereas the latter cytoplasmic.

Age Factors↗
PubMed1999

Basonuclin is associated with the ribosomal RNA genes on human keratinocyte mitotic chromosomes.

Basonuclin is a zinc finger protein mainly expressed in keratinocytes of the basal layer of epidermis and the outer root sheath of hair follicles. It is also found in abundance in the germ cells of testis and ovary. In cultured keratinocytes, basonuclin is associated with chromatin in all phases of the cell cycle, including mitosis. By immunocytochemical methods, we demonstrate here that in mitosis basonuclin is associated with the short arms of the acrocentric chromosomes and with other loci on many metaphase chromosomes of human keratinocytes. Using the evolutionarily highly conserved N-terminal pair of zinc fingers in an electrophoresis mobility shift assay, we demonstrate that the DNA target sequences of basonuclin on the acrocentric chromosomes are likely to be within the promoter region of the 45S rRNA gene transcription unit. DNase I footprinting shows that basonuclin zinc fingers interact with the upstream control element of this promoter, which is necessary for the high level of transcription of the rRNA genes. This result suggests that basonuclin may be a tissue-specific transcription factor for the ribosomal RNA genes.

Base Sequence↗
PubMed1999

DNA cloning without restriction enzyme and ligase.

One common problem in using the traditional DNA cloning procedure is that suitable natural restriction sites are often unavailable for a given task. Creating new restriction sites is often time consuming. Here, I describe a simple technique of producing "customized cohesive ends" by a combination of PCR primer design and lambda exonuclease digestion. These complementary cohesive ends can form hybrids to link two sequences. Because the overhangs created by lambda exonuclease are slightly longer than the complementary sequence, after hybrid formation, a stretch of single-strand gap remains, which then is repaired by Klenow (3'-->5' exo-) enzyme. The repair process also stabilizes the linkage. Because of the independence from natural or artificial restriction sites, this method allows rapid and precise insertion of one DNA fragment into another at virtually any position. It also simplifies the planning of a cloning strategy, increases recombinant frequency and is suitable for automation.

Base Sequence↗
PubMed1998

Basonuclin, a zinc finger protein associated with epithelial expansion and proliferation.

Basonuclin is a zinc finger protein first described as a keratinoycte specific protein present in various stratified squamous epithelia found in epidermis, esophagis, cornea and virgina. Recent studies showed that its distribution also includes the germ cells of testis and ovary. The presence of basonuclin appeared always related to the cellular proliferative ability not just cell division, for it was found both in dividing and quiescent cells. Basonuclin disappeared when a cell became post-mitotic. This review examines the latest findings about the distribution, molecular and cellular biology of basonuclin and discusses its possible role in cell proliferation.

Animals↗
PubMed1998

Translocation of the zinc finger protein basonuclin from the mouse germ cell nucleus to the midpiece of the spermatozoon during spermiogenesis.

Basonuclin was first described as a human keratinocyte zinc finger protein present in the nuclei of proliferative basal keratinocytes in the epidermis. It disappears from keratinocytes that have lost their proliferative ability and have entered terminal differentiation. We now report that basonuclin is present also in the germ cells of the mouse testis and ovary. Immunocytochemical staining detected basonuclin in the nuclei of spermatogonia and spermatocytes at various developmental stages. During spermiogenesis, it relocated from the nucleus to the midpiece of the flagellum of the spermatozoa. In the ovary, basonuclin was found mainly in the nuclei of developing oocytes. The dual presence of basonuclin in differentiated spermatozoa and oocytes suggests that it may play a role in their differentiation and the early development of an embryo.

Animals↗
PubMed1997

Complementary oligonucleotides and the origin of the mammalian involucrin gene.

A large portion of the consensus sequence of four mammalian involucrin genes comprises four pairs of complementary oligonucleotides. One of the oligonucleotides in each pair is rich in purines and the other in pyrimidines. The perfect complementarity of a number of pairs apparently resulted from a concerted single nucleotide substitution in each pairing oligonucleotide from their respective consensus. This suggests that one member of the pair was derived from the other member of the same pair by a template-dependent replication mechanism. Such a replication mechanism differs from the one that creates tandem repeats by producing short complementary sequence on the same strand of DNA.

Amino Acid Sequence↗
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↗
PubMed1995

Protein kinases as mediators of fluid shear stress stimulated signal transduction in endothelial cells: a hypothesis for calcium-dependent and calcium-independent events activated by flow.

Fluid shear stress regulates endothelial cell function, but the signal transduction mechanisms involved in mechanotransduction remain unclear. Recent findings demonstrate that several intracellular kinases are activated by mechanical forces. In particular, members of the mitogen-activated protein (MAP) kinase family are stimulated by hyperosmolarity, stretch, and stress such as heat shock. We propose a model for mechanotransduction in endothelial cells involving calcium-dependent and calcium-independent protein kinase pathways. The calcium-dependent pathway involves activation of phospholipase C, hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2), increases in intracellular calcium and stimulation of kinases such as calcium-calmodulin and C kinases (PKC). The calcium-independent pathway involves activation of a small GTP-binding protein and stimulation of calcium-independent PKC and MAP kinases. The calcium-dependent pathway mediates the rapid, transient response to fluid shear stress including activation of nitric oxide synthase (NOS) and ion transport. In contrast, the calcium-independent pathway mediates a slower response including the sustained activation of NOS and changes in cell morphology and gene expression. We propose that focal adhesion complexes link the calcium-dependent and calcium-independent pathways by regulating activity of phosphatidylinositol 4-phosphate (PIP) 5-kinase (which regulates PIP2 levels) and p125 focal adhesion kinase (FAK, which phosphorylates paxillin and interacts with cytoskeletal proteins). This model predicts that dynamic interactions between integrin molecules present in focal adhesion complexes and membrane events involved in mechanotransduction will be integrated by calcium-dependent and calcium-independent kinases to generate intracellular signals involved in the endothelial cell response to flow.

Calcium↗
PubMed1995

Fluid shear stress stimulates mitogen-activated protein kinase in endothelial cells.

Local alterations in the hemodynamic environment regulate endothelial cell function, but the signal-transduction mechanisms involved in this process remain unclear. Because mitogen-activated protein (MAP) kinases have been shown to be activated by physical forces, we measured the phosphorylation and enzyme activity of MAP kinase to identify the signal events involved in the endothelial cell response to fluid shear stress. Flow at physiological shear stress (3.5 to 117 dynes/cm2) activated 42-kD and 44-kD MAP kinases present in cultured bovine aortic endothelial cells, with maximal effect at 12 dynes/cm2. Activation of a G protein was necessary, as demonstrated by complete inhibition by the nonhydrolyzable GDP analog GDP-beta S. Activation of protein kinase C (PKC) was required, as shown by inhibiting PKC with staurosporine or downregulating PKC with phorbol 12,13-dibutyrate. Both Ca(2+)-dependent and -independent PKC activity, measured by translocation and substrate phosphorylation, increased in response to flow. However, MAP kinase activation was not dependent on Ca2+ mobilization, since Ca2+ chelation had no inhibitory effect. On the basis of these findings, it is proposed that flow activates two signal-transduction pathways in endothelial cells. One pathway is Ca2+ dependent and involves activation of phospholipase C and increases in intracellular Ca2+. A new pathway, described in the present study, is Ca2+ independent and involves a G protein and increases in PKC and MAP kinase activity.

Alkaloids↗
PubMed1994

Association of basonuclin with ability of keratinocytes to multiply and with absence of terminal differentiation.

Basonuclin is a protein possessing three pairs of zinc fingers and a nuclear localization signal. Expression of the gene is largely confined to keratinocytes of stratified squamous epithelia and hair follicles. In the epidermis and in stratified epidermal cultures, basonuclin is present in the nuclei of cells in or close to the basal layer but not in the nuclei of cells in more superficial layers. The Ki-67 protein, a nuclear marker for any stage of the multiplication cycle is present in only a subclass of basonuclin-containing cells. In cultured keratinocytes, the disappearance of basonuclin mRNA is associated with loss of colony-forming ability and the appearance of mRNA for involucrin, a protein characteristic of terminal differentiation. In hair follicles, the largest reservoir of basonuclin-containing cells is the outer root sheath, which contains precursors of differentiated cells of the hair shaft and of the epidermis. Basonuclin is not a cell cycle marker but is likely instead to be a regulatory molecular whose presence in the keratinocyte is linked to the maintenance of proliferative capacity and prevention of terminal differentiation.

Adult↗
PubMed1993

Olfactory memory in unmedicated schizophrenics.

Previous studies have indicated that schizophrenic patients have olfactory deficits. The question as to whether olfactory deficits are due to chronic effects of medication has not been addressed. This is the first paper to report that never-medicated schizophrenic patients also have olfactory deficits. Twenty four normal subjects and twenty unmedicated schizophrenic patients were examined with two tests of olfactory function: the University of Pennsylvania Smell Identification Test (UPSIT) and a match-to-sample olfactory memory test. Results indicated that schizophrenics did poorly on both the UPSIT and the olfactory match-to-sample memory test relative to sex and age-matched controls. ANCOVA showed that the deficit in performance on the olfactory match-to-sample test was still present even when the variance due to the UPSIT was taken out of the analysis. Deficits in olfactory identification and olfactory memory are consistent with the concept that schizophrenics have dysfunctional limbic systems.

Blood Glucose↗
PubMed1992

Basonuclin: a keratinocyte protein with multiple paired zinc fingers.

A cDNA clone has been prepared from mRNA of cultured human keratinocytes. The sequence of the cDNA reveals that in the C-terminal two-thirds of the corresponding protein (basonuclin), there are three separated pairs of adjacent zinc fingers. The amino acid sequence of each pair is homologous to that of the single pair of zinc fingers of the Drosophila transcription factor encoded by disco. Near the C-terminal end of basonuclin and on the surface of a putative alpha-helix, there is a stripe of serine residues similar to that of the transcription factor PRDII-BF1. Basonuclin possesses a sequence of six amino acids quite similar to one present in the myogenic family of proteins, including Myf5; this sequence is located in the omega loop of the myogenic proteins but within a zinc finger of the keratinocyte protein. As basonuclin is present mainly in the nuclei of the basal cell layer, its regulatory function is likely to be exerted prior to the process of terminal differentiation.

Amino Acid Sequence↗
PubMed1992

The Na/K/2Cl cotransporter is increased in hypertrophied vascular smooth muscle cells.

Hypertrophy of vascular smooth muscle cells (VSMC) is a pathogenic feature of hypertension which may contribute to abnormal vessel tone and function. As a consequence of the increase in cell size associated with hypertrophy, it is likely that alterations in the mechanisms that regulate VSMC intracellular volume occur. Because the Na+/H+ exchanger plays an important role in volume regulation and because we previously observed long term alterations in Na+/H+ exchange and pHi in response to angiotensin-II-induced (ang II) hypertrophy, we studied cell-acidifying mechanisms. To do this, we measured alkaline recovery from NH4Cl-mediated alkalinization, using the fluorescent dye, 2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein. VSMC were growth-arrested (0.4% calf serum for 24 h) or hypertrophied (100 nM ang II in 0.4% calf serum for 24 h). Ang II-treated cells exhibited a 107% increase in alkaline recovery over control cells (13.86 +/- 1.87 versus 6.68 +/- 1.01 mmol H+/min/liter cells). The increase in alkaline recovery was not a result of increased Cl-/HCO-3 exchange becaue it was not HCO-3 dependent nor inhibited by 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid. Studies with bumetanide and the sterically inhibited substrate N(CH3)4+ showed that the alkaline recovery was mediated by NH4+ transport via the Na/K/2Cl cotransporter. Ang II-treated cells exhibited a 334% increase in bumetanide-sensitive alkaline recovery over control cells (9.16 +/- 1.90 versus 2.11 +/- 1.46 mmol H+/min/liter cells). Ang II-treated cells also exhibited a 90% increase in bumetanide-sensitive 86Rb uptake over control cells. These findings demonstrate that Na/K/2Cl cotransport activity is specifically induced in ang II-hypertrophied VSMC and establish this transporter as a component of the hypertrophic growth response.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗