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D Lin

Publications and source records attributed to D Lin.

At least 181 records · Page 10Linked to original sources

A preliminary study of the fixation mechanism of collagen reaction with a polyepoxy fixative.

A new biomaterial has been developed by fixing native collagens with a polyepoxy compound (PC) fixative. Prior studies have shown that this biomaterial has comparable properties as compared to collagen fixed with glutaraldehyde (GA) and thus has a great promise for use as an implantable bioprosthesis. The purpose of this study was to understand the mechanism of the amino acids-PC reactions in the fixation process. Bovine arteries were fixed with PC under various pH, concentration and temperature conditions as a function of fixation time. Individual amino acid components in the fresh and the fixed arteries were assayed using a Beckman amino acid analyzer to determine the degree of tanning. The denaturation temperature (Td) was also measured on each sample. Since the denaturation temperature is a direct indication of cross-linking of individual amino acids with the fixative, the difference in the degree of tanning for the same increase in Td may be indicative of the quantity of the masked, non-cross-linked amino acids. The fixation reaction data indicated that not all amino acids were cross-linked upon contacting the PC fixative. Masking appeared to be more substantial with a fixation at higher pH values.

Amino Acids↗

Lipid accumulation and body fat distribution is influenced by type of dietary fat fed to rats.

The amount of fat in the diet is known to influence body weight and body composition, but it is not clear whether dietary fat composition can affect body composition independently. We studied the effects of six months of feeding diets containing lard (L), corn oil (CO), fish oil (FO) or medium chain triglycerides (MCT) on body weight and body composition in adult male Wistar rats. Although FO fats ate slightly less total energy than the other groups, there were no differences among groups in body weight at any time during the study. However, body composition, the composition of depot triglyceride, body fat distribution and insulin resistance were all influenced by the type of fat in the diet. FO rats had less total body fat, less intra-abdominal fat, and less insulin resistance than all other groups. Although some of these metabolic effects may have been secondary to a slightly lower energy intake, we believe these data demonstrate the potential impact which dietary fat composition can have on metabolism and body weight regulation.

Adipose Tissue↗

Mutation of the serine 15 phosphorylation site of human p53 reduces the ability of p53 to inhibit cell cycle progression.

Overexpression of wild-type p53 prevents cells from entering the S phase of the cell cycle. The amino-terminal transactivation region of p53 is phosphorylated by several protein kinases, including DNA-PK, a nuclear serine/threonine protein kinase that in vitro requires DNA for activity. DNA-PK was recently shown to phosphorylate serines 15 and 37 of human p53 (Lees-Miller et al., 1992. Mol. Cell. Biol., 12, 5041-5049). To prevent phosphorylation at these sites, mutants were constructed that changed the codons for serine 15 or serine 37 to alanine codons. Expression of p53-Ala-37 in stably transformed T98G cells blocked progression of the cells into S phase as well as did the expression of wild-type p53. In contrast, p53-Ala-15 was partially defective in blocking cell cycle progression. Several cell clones transformed with the mutant p53-Ala-15 gene expressed normal levels of p53 mRNA but accumulated little or no detectable p53 protein. However, by using a transient expression system driven by a strong cytomegalovirus promoter, we showed that the inability of p53-Ala-15 to fully block cell cycle progression was not due to inadequate levels of expression or to a failure of the mutant protein to accumulate in the nucleus. These results suggest that phosphorylation of Ser-15 may affect p53 function.

Amino Acid Sequence↗

Cell cycle effects of microinjected antisense oligodeoxynucleotides to p34cdc2 kinase.

In this study the effect of antisense oligomers targeted against the mRNA transcripts of p34cdc2 kinase on G1 progression into S-phase was examined. For this purpose, antisense, sense, or nonsense oligomers were introduced directly into the cytoplasm of T98G cells grown in monolayer cultures by glass-capillary microinjection. The microinjection of antisense oligomers (but not sense or nonsense oligomers) into growth-arrested cells before serum stimulation inhibited G1 progression into S-phase. This inhibition was correlated with a reduction in the steady-state levels of nuclear p34cdc2 protein. Microinjection of antisense oligomers into cells at 2 and 6 hours after serum stimulation also resulted in a marked inhibition in the ability of cells to enter S-phase. The inhibitory effect decreased when cells were microinjected at 12 hours after serum stimulation. When cells were microinjected at 18 and 24 hours after serum stimulation, only a slight inhibition was observed. As the antisense oligomers were introduced directly into the cytoplasm of cells at each of the time points examined, the observed differences in the inhibitory effects of the antisense oligomers at later times after serum stimulation cannot be explained by differences in uptake. An alternative explanation is that after a certain threshold level of nuclear p34cdc2 protein is reached in late G1 phase; no further increase is necessary, because the cells become committed to enter S-phase. In yeast, p34cdc2 appears to play an important role in the G1/S-phase transition at a control point in late G1 phase called START (reviewed by Lewin). In mammalian cells a control point that could be equivalent to START is the "restriction point" which is defined as the time after which inhibition of protein synthesis fails to block entry into S-phase (reviewed by Pardee). The effects observed with antisense oligomers to p34cdc2 kinase are strikingly similar to what is observed when low concentrations of the drug cycloheximide are added to these cells at different times after serum stimulation; entry into S-phase is significantly inhibited when cycloheximide is added up to 12 hours postimulation. Thus, the results reported in this study are in agreement with the idea that p34cdc2 kinase plays a role in the G1/S phase transition in mammalian cells. Finally, introduction of antisense oligomers directly into the cytoplasm of cells grown in monolayer cultures by glass-capillary microinjection appears to be a viable alternative to simply adding the oligomers to the culture medium.(ABSTRACT TRUNCATED AT 400 WORDS)

Base Sequence↗

Growth arrest induced by wild-type p53 protein blocks cells prior to or near the restriction point in late G1 phase.

Conditional expression of wild-type (wt) p53 protein in a glioblastoma tumor cell line has been shown to be growth inhibitory. We have now more precisely localized the position in the cell cycle where growth arrest occurs. We show that growth arrest occurs prior to or near the restriction point in late G1 phase of the cell cycle. The effect of wt p53 protein on the expression of four immediate-early genes (c-FOS, c-JUN, JUN-B, and c-MYC), one delayed-early gene (ornithine decarboxylase), and two late-G1/S-phase genes (B-MYB and DNA polymerase alpha) was also examined. Of this subset of growth response genes, only the expression of B-MYB and DNA polymerase alpha was significantly repressed. The possibility that decreased expression of B-MYB may be an important component of growth arrest mediated by wt p53 protein is discussed.

Blotting, Northern↗

Lin et al. reply.

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Journal Article↗

cAMP post-transcriptionally diminishes the abundance of adrenodoxin reductase mRNA.

Adrenodoxin reductase (AR; ferridoxin: NADP+ oxidoreductase, EC 1.18.1.2) is a flavoprotein that mediates electron transport from NADPH to all known mitochondrial forms of cytochrome P450. AR mRNA was found in all human adult and fetal tissues examined; however, it was vastly more abundant in tissues that synthesize steroid hormones. The ratio of the 18- form of mRNA lacking 18 alternately spliced bases to the 18+ form was approximately 100:1 and remained constant irrespective of the tissue or hormonal manipulation, indicating that the alternate splicing is a passive nonregulated event. AR protein was unchanged by forskolin treatment of human JEG-3 cytotrophoblast cells for 24 h, but the mRNA diminished. Phorbol 12-myristate 13-acetate and cycloheximide had no effect, even though these agents had the expected effects on P450scc and adrenodoxin mRNAs. cAMP decreased the abundance of AR mRNA expressed from both transfected plasmids and the endogenous gene, indicating the effect was post-transcriptional. AR gene transcription in JEG-3 cells and promoter-chloramphenicol acetyltransferase constructs transfected into JEG-3 cells were unresponsive to forskolin. Powerful basal transcription elements were identified between -46 and -214 bases from the principal transcriptional initiation site, a region containing six elements closely resembling the binding site for transcription factor SP1.

Cells, Cultured↗

Genotypic identification and characterization of species and strains within the genus Candida by using random amplified polymorphic DNA.

Random amplified polymorphic DNA (RAPD) was used to better characterize the genotypic relatedness among medically important Candida species. By using short oligomer primers (10-mers) with arbitrarily chosen sequences in the polymerase chain reaction, distinctive and reproducible sets of polymerase chain reaction products were observed for isolates of C. albicans, C. lusitaniae, C. tropicalis, and Torulopsis (Candida) glabrata. The RAPD analysis differentiated a physiologically homogeneous panel of C. parapsilosis into three distinct groups and showed genetic diversity within C. haemulonii. Intraspecies DNA-length polymorphisms were seen for RAPD profiles derived from different isolates of each species. Analysis of RAPDs from a panel of C. albicans, which included 16 laboratory derivatives of two reference strains, showed that the profiles of unrelated strains differed and that the derivatives of each reference strain were identifiable. Minor differences in the RAPD profiles, suggestive of mutations that had occurred during the long-term maintenance of the strains, were detected. Because of its ease and reliability, RAPD analysis should be useful in providing genotypic characters for taxonomic descriptions, for confirming the identities of stock isolates, for typing Candida species in epidemiologic investigations, and for use in the rapid identification of pathogenic fungi.

Base Sequence↗

Skeletal muscle composition in dietary obesity-susceptible and dietary obesity-resistant rats.

The intent of this study was to determine whether a relationship exists between susceptibility to high-fat diet (HFD)-induced obesity and skeletal muscle fiber type. Forty-four adult male Wistar rats were given ad libitum access to a HFD (60% of calories from fat) for 4 wk. Rats were then grouped into quartiles for total weight gain, and the top-quartile [obesity prone (OP)] rats were compared with the bottom-quartile [obesity resistant (OR)] rats. OP rats gained 1.5 times as much weight as OR rats. OR rats had a significantly higher proportion of type I muscle fibers in the medial head of the gastrocnemius muscle than OP rats both before (determined from a muscle biopsy) and after the HFD feeding period. A greater proportion of type I fibers may be associated with a greater capacity for fat oxidation, which would favor resistance to body fat accumulation. Preexisting differences in muscle fiber composition may play a role in determining susceptibility to dietary obesity.

Animals↗

[Prognostic significance of nuclear DNA measurement with flow cytometry for laryngeal squamous cell carcinomas].

DNA ploidy, S% and proliferative index (PI) of carcinomatous cell were evaluated with FCM in 41 patients with laryngeal primary squamous cell carcinomas which were operated on in our department during the period between Jan. 1979 to Oct. 1987. The results showed that DNA aneuploidy was identified in 65.85% of the cases (27/41) and patients with aneuploid carcinomas had higher S% (mean +/- standard: 23.22 +/- 9.57%), higher PI (mean +/- standard: 37.59 +/- 9.57%) and higher rate of relapse and/or metastasis than those with diploid tumours (55.55% vs 11.77%, P < 0.025). Patients with aneuploid carcinomas also correlated with an unfavorable prognosis compared with those with diploid tumors. Cases with aneuploid and diploid carcinomas had 66.11% and 92.86% respectively for two years postoperation survival rate; 46.74% and 92.86% respectively for five years postoperation survival rate (Log-rank test: chi 2 = 5.96, P < 0.025). We conclude that nuclear DNA measurement with flow cytometry is of important significance in estimating prognosis and guiding treatment for laryngeal squamous cell carcinomas.

Aged↗

Development of dietary obesity in rats: influence of amount and composition of dietary fat.

This study was conducted to examine long-term effects of amount and type of dietary fat on body weight and body composition. Adult male Wistar rats were fed high fat (HF; 60% of calories) or low fat (LF; 20% of calories) diets for 28 weeks. Half of the rats in each condition received diets with saturated fat (lard) (S) and the remainder received diets with polyunsaturated fat (corn oil) (U). From 28-39 weeks, HF rats were switched to LF diets (fat type remained constant). From 40-50 weeks, previously HF fed rats were weight-matched to rats in the LF fed groups. HF rats became fatter than LF rats during weeks 1-28 and remained heavier and fatter from weeks 28-39. During weeks 1-28, type of dietary fat had no effect on total body fat in either HF or LF rats, but during period 2 (weeks 28-39), U rats were heavier and fatter than S rats. There was some indication that U diets were associated with greater accumulation of fat in subcutaneous adipose tissue depots than S diets. From 40-50 weeks, rats previously fed the HF diet required less food to maintain their body weight than did LF diet rats. In summary, these results suggest that although both amount and type of dietary fat can affect body weight and body composition, the effects of the type of fat are less than those of amount of dietary fat.

Adipose Tissue↗

[Estrogen and progesterone receptors in laryngeal carcinoma].

The existence of hormone receptors on or within neoplastic tissue has potential diagnostic, therapeutic, and prognostic importance. It has now been demonstrated in many reports that cancers of the breast and prostate often express hormone receptors and can be controlled by hormone manipulation. The fact that larynx is a target organ for androgenic steroids has long been known. The question of whether laryngeal carcinoma expresses hormone receptors has been a matter of interest. We adopted the fluorescent hormone conjugate method to measure the estrogen receptor (ER) and progesterone receptor (PR) in 25 patients with laryngeal cancer. Our findings indicated that in 25 samples of laryngeal cancer 48% were ER positive, and 68% were PR positive. Cells of the laryngeal cancer not only contain ER and PR but also have relationship between the presence of receptors and the degree of histologic differentiation. Low differentiated cancers are inclinable to contain low level receptors. From this initial survey it appears that the hormone receptors could play an important role in the cancer of larynx. The presence of receptors in some laryngeal carcinoma implicates that these tumors are possibly hormone sensitive.

Adult↗