Biomedical subjects

K Yang

Publications and source records attributed to K Yang.

At least 37 records · Page 2Linked to original sources

PubMed1996

Epithelial cell hyperproliferation induced in the exocrine pancreas of mice by a western-style diet.

BACKGROUND: Pancreatic cancer is a common cause of mortality in the United States, with an estimated 27,800 people dying of the disease in this country in 1996. Epidemiologic studies have suggested that Western diets containing high fat, high protein, and low calcium contents are associated with increased incidence of pancreatic cancer. PURPOSE: We investigated whether a Western-style diet containing increased fat content and decreased calcium and vitamin D contents would induce epithelial cell hyperproliferation (excess cell duplication) or hyperplasia (excess cell accumulation) in the pancreas, as was previously demonstrated in the colon and mammary gland. METHODS: C57BL/6J mice at 4 weeks of age were randomly assigned to one of two groups of 14 mice each. One group received the control diet ad libitum, and the other group was given the Western-style diet ad libitum. After 6, 9, and 15 weeks on the diet, four or five mice per group were infused with 5-bromo-2'-deoxyuridine (BrdU) for 72 hours by use of subcutaneously implanted Alzet osmotic pumps. The mice were then killed, and the pancreas of each mouse was removed. In the exocrine pancreas with ductal secretion, the duct system (including interlobular and intralobular ducts and centroacinar [i.e., centroductular] cells) and acini were measured both histopathologically and immunohistochemically (BrdU) and were analyzed without knowledge of the source of the specimens. Two-way analysis of variance was carried out. All P values were generated from two-sided tests for statistical significance. RESULTS: The number of pancreatic ducts (interlobular, intralobular, and centro-acinar-cancer-prone regions in certain rodent models and in humans) and acini per mouse in the Western-style diet group was similar to that in the control diet group during the entire feeding period (P = .76, .32, .93, and .42, respectively). Statistically significant higher BrdU-labeling indices of the ductal interlobular and intralobular epithelial cells were seen in mice fed the Western-style diet than in mice fed the control diet during the entire observation period (P = .014 and .016, respectively). There was no statistically significant difference (P = .098) between both diet groups in the BrdU-labeling indices of the centroacinar epithelial cells. CONCLUSIONS: A Western-style diet induced pancreatic epithelial cell hyperproliferation in mice, further suggesting that increased fat content and decreased calcium and vitamin D contribute to the development of pancreatic neoplasms.

Animals↗

PubMed1996

Requirement of rigid-body motion of transmembrane helices for light activation of rhodopsin.

Conformational changes are thought to underlie the activation of heterotrimeric GTP-binding protein (G protein)-coupled receptors. Such changes in rhodopsin were explored by construction of double cysteine mutants, each containing one cysteine at the cytoplasmic end of helix C and one cysteine at various positions in the cytoplasmic end of helix F. Magnetic dipolar interactions between spin labels attached to these residues revealed their proximity, and changes in their interaction upon rhodopsin light activation suggested a rigid body movement of helices relative to one another. Disulfide cross-linking of the helices prevented activation of transducin, which suggests the importance of this movement for activation of rhodopsin.

Amino Acid Sequence↗

PubMed1996

Apoptosis, cell replication, and Western-style diet-induced tumorigenesis in mouse colon.

In this study, feeding Western-style diets (WDs) to mice for a duration of two years without any chemical carcinogen led to the development of gross colonic lesions that were histologically classified as dysplastic crypts and focal hyperplasias with or without atypical nuclei. To better understand early biological events contributing to the development of colonic neoplasia, grossly normal colonic mucosa was investigated; mitotic and apoptotic colonic epithelial cells, atypical mitosis, and atypical nuclei were studied. A significant and transient increase of mitotic activity in the basal and intermediate portions of the colonic crypts was seen in young mice after feeding them the WDs. This was accompanied by diffuse activation of apoptosis of the colonic epithelial cells. In the middle of the rodents' life span, after administration of both the WDs and control diet, the rodents developed a marked depletion of apoptotic epithelial cells in the mid-region of the colonic crypts; this was followed by the expansion of an epithelial cell population containing atypical nuclei, and the emergence of the gross lesions noted above. With this sequence of events, prolonged feeding of WDs to mice produced single-crypt dysplastic lesions and focal hyperplasias indicative of tumorigenesis.

Animals↗

PubMed1996

Cytokeratin, lectin, and acidic mucin modulation in differentiating colonic epithelial cells of mice after feeding Western-style diets.

Several studies have recently reported the development of colonic epithelial cell hyperproliferation in rodents following the ingestion of Western-style diets. In this study, additional measurements related to differentiation and maturation of the colonic epithelial cells were made after feeding this type of diet. Two Western-style diets high in fat and phosphate content and low in calcium and vitamin D were fed to C57BL/6J mice for 12, 24, and 52 weeks. Diet A contained American Blend fat as a source of lipids, diet B contained corn oil, and control diet C was a standard AIN-76A semisynthetic diet which is lower in fat content and higher in calcium and vitamin D. Colonic epithelial cells were studied for three biomarkers: cytokeratin catalogue no. 18 (clone LE64) expression, soybean agglutinin carbohydrate lectin binding, and acidic mucins including sialo- and sulfomucins. Feeding of diets A and B revealed that colonic epithelial cells had increased expression of cytokeratin catalogue 18 and SBA carbohydrate lectin binding compared to controls (P = 0.0001 for diet A versus C and diet B versus C). Significant differences were found between diets B and C (P = 0.0001) and diets A and C (P = 0.0001) in total acidic mucins and in the ratio of sialomucin:sulfomucin (P = 0.0001). These findings demonstrate that both functional and structural modifications occurred in colonic epithelial cells under these dietary conditions, and further defined this rodent model for preclinical evaluation of nutritional and chemopreventive interventions.

Animals↗

PubMed1996

Structural and physiologic characterization of the mid-region secretory species of parathyroid hormone-related protein.

Parathyroid hormone-related protein (PTHrP) is initially translated as a preprohormone which is posttranslationally processed to yield a family of mature secretory forms. Most attention has focused on the amino-terminal portion of the molecule which is homologous to parathyroid hormone. It is clear, however, that a mid-region species of PTHrP is posttranslationally cleaved from the highly conserved mid-region of PTHrP, and that the amino terminus of this peptide is Ala38. The purposes of the current study were three: 1) to confirm that Arg37 immediately preceding Ala38 serves as a posttranslational processing site in the PTHrP precursor, 2) to determine the carboxyl terminus of the mid-region secretory species of PTHrP, and 3) to synthesize this authentic mid-region secretory form of PTHrP and determine whether it is biologically active. The results indicate that: 1) Arg37 is indeed a processing site in the PTHrP precursor; 2) three distinct mid-region PTHrP species are generated by posttranslational processing, PTHrP(38-94)amide, PTHrP(38-95), and most likely, PTHrP(38-101); and 3) synthetic mid-region PTHrP(38-94)amide is active in four different biological systems. These studies confirm the finding that PTHrP is a prohormone. More importantly, they define a novel, biologically active highly conserved mid-region secretory form of PTHrP.

Amino Acid Sequence↗

PubMed1996

Structure and function in rhodopsin. Single cysteine substitution mutants in the cytoplasmic interhelical E-F loop region show position-specific effects in transducin activation.

The cytoplasmic interhelical E-F loop in rhodopsin is a part of the region that interacts with the G-protein transducin and rhodopsin kinase during signal transduction. In extending the previous work on systematic single cysteine substitutions of the amino acids in the cytoplasmic C-D loop, we have now replaced, one at a time, the amino acids Q225-I256 in the E-F loop region by cysteines. All the mutants formed the characteristic rhodopsin chromophore with 11-cis-retinal. While most of the mutants bleached normally, L226C, showed abnormal bleaching behavior. A study of the alkylation of the mutants by N-ethylmaleimide in dark showed low reactivity by some mutants, especially L226C. The rates of transducin activation (GT(alpha)-GTP gamma S complex formation) were measured for all the mutants. While these were normal for the bulk of the mutants, some (L226C, T229C, V230C, A233C, A234C, T242C, T243C, and Q244C) showed strikingly reduced transducin activation. The results suggest a specific structure in the E-F loop that interacts with transducin.

Amino Acid Sequence↗

PubMed1996

Structural features and light-dependent changes in the cytoplasmic interhelical E-F loop region of rhodopsin: a site-directed spin-labeling study.

Thirty consecutive single cysteine substitution mutants in the amino acids Q225-I256 of bovine rhodopsin have been prepared and modified with a sulfhydryl specific nitroxide reagent. This sequence includes the E-F interhelical loop, a transducin interaction site. The accessibilities of the attached nitroxides to collisions with hydrophilic and hydrophobic paramagnetic probes in solution were determined, and the electron paramagnetic resonance spectra analyzed in terms of side chain mobility, both in the dark and after photoactivation. Accessibility cata shows that the rhodopsin polypeptide chain crosses an aqueous/ hydrophobic boundary in the range V227-K231 and again in the range V250-V254. In the hydrophobic segments, both the accessibility and mobility data are consistent with helical structures. In the regions of the sequence located within the aqueous phase, periodic variation in both accessibility and mobility of the spin-labeled side chains indicates that the E-F interhelical loop is largely alpha-helical, being formed by regular extensions of the E and F helices by about 1.5 and 3 turns, respectively. Judging from nitroxide mobilities, the putative extension of helix E in the aqueous phase is more dynamic than that of helix F. Changes in the electron paramagnetic resonance characteristics of the spin-labeled rhodopsin upon photoactivation indicate that chromophore isomerization results in patterns of structural changes that can be interpreted in terms of movements of helices that extend into the aqueous loop regions.

Amino Acid Sequence↗

PubMed1996

Novel receptor mechanisms for heroin and morphine-6 beta-glucuronide analgesia.

The rapid metabolism of heroin to 6-acetylmorphine and its slower conversion to morphine has led many to believe that heroin and morphine act through the same receptors and that the differences between them are due to their pharmacokinetics. We now present evidence strongly implying that heroin and two potent mu drugs, fentanyl and etonitazine, act through a unique receptor mechanism similar to morphine-6 beta-glucuronide which is readily distinguished from morphine. Heroin, 6-acetylmorphine and morphine-6 beta-glucuronide show no analgesic cross tolerance to morphine in a daily administration paradigm, implying distinct receptors. Strains also reveal analgesic differences among the drugs. CXBK mice, which are insensitive to morphine, retain their analgesic sensitivity to heroin, 6-acetylmorphine, morphine-6 beta-glucuronide, fentanyl and etonitazine. Antisense mapping of the mu opioid receptor MOR-1 reveals that oligodeoxynucleotide probes against exon 2, which are inactive against morphine analgesia, block morphine-6 beta-glucuronide, heroin, fentanyl and etonitazine analgesia. Finally, an antisense probe targeting Gi alpha 1 blocks both heroin and morphine-6 beta-glucuronide, but not morphine, analgesia. These results indicate that heroin, 6-acetylmorphine, fentanyl and etonitazine all can produce analgesia through a novel mu analgesic system which is similar to that activated by morphine-6 beta-glucuronide.

Analgesics↗

PubMed1996

Aromatic DNA adducts in lymphocytes of humans working at high and low traffic density areas.

Aromatic DNA adduct levels were determined by the 32P-postlabelling assay in lymphocytes isolated from newspaper vendors working at urban high traffic areas (n = 31) and suburban low traffic areas (n = 22) in Milan, Italy. The DNA adduct levels ranged from 0.7 to 6.7/10(8) nucleotides, while most of them were between 1.0 and 3.0/10(8) nucleotides. No difference was found between the DNA adduct levels of the high-exposed group (2.2/10(8) and the low-exposed group (2.2/10(8). The heavy smokers (n = 8) had 23% higher DNA adduct level (2.7/10(8)) than the non-smokers (n = 37, 2.2/10(8) (P = 0.27), but no correlation was found between the adduct level and the number of cigarettes/day. Analysis of variance of the DNA adduct levels among the 14 pairs of individuals working at the same news-stands revealed little effect of the environmental air exposure on the DNA adduct level.

DNA Adducts↗

PubMed1996

Ovine 11 beta-hydroxysteroid dehydrogenase type 2 gene predicts a protein distinct from that deduced by the cloned kidney cDNA at the C-terminus.

The gene encoding ovine 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD2) was cloned and characterized. This gene consists of five exons and is greater than 4 kb in length. It contains an open reading frame of 1215 bp, which encodes a protein of 404 amino acids with a predicted MW of 44 kDa. The deduced ovine 11 beta-HSD2 protein displays over 78% sequence identity to those of the human, rabbit, rat, and mouse. However, this differs from the published sequence of ovine kidney 11 beta-HSD2 cDNA which predicts a protein of 427 amino acids. Sequence alignment indicated that this discrepancy is attributed to two single nucleotide omissions in the published cDNA sequence which resulted in a shift in the open reading frame at the codon for residue 358. Therefore, the present results have provided conclusive evidence that the primary structure of 11 beta-HSD2 protein is well conserved between the sheep and the other four mammals. Moreover, Northern blot analysis of total RNA samples from 15 peripheral tissues and seven brain regions of the mature fetal sheep revealed that the expression of 11 beta-HSD2 gene is highly tissue-specific in that it is only expressed in the kidney and adrenal gland, and at a much lower abundance in the testis, colon and placenta. The cloning of the sheep 11 beta-HSD2 gene should facilitate future studies on the regulation of 11 beta-HSD2 gene expression during fetal development in a mammalian model.

11-beta-Hydroxysteroid Dehydrogenases↗

PubMed1996

Increased expression of brain-derived neurotrophic factor but not neurotrophin-3 mRNA in rat brain after cortical impact injury.

Levels of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3) mRNA expression were measured in a rodent model of traumatic brain injury (TBI) following unilateral injury to the cerebral cortex. To obtain reliable data on the co-expression of neurotrophin genes, adjacent coronal sections from the same rat brains were hybridized in situ with BDNF and NT3 cRNA probes. BDNF mRNA increased at 1,3, and 5 hr after unilateral cortical injury in the cortex ipsilateral to the injury site and bilaterally in the dorsal hippocampus. NT3 mRNA did not change significantly following injury. Our results suggest that TBI produces rapid increases in BDNF mRNA expression in rat brain without changes in NT3 mRNA expression, a finding which differs from studies of ischemia and seizures. It is possible that increased levels of BDNF mRNA rather than NT3 are important components of pathophysiological responses to TBI.

Analysis of Variance↗

PubMed1996

Liposome-mediated NGF gene transfection increases ChAT activity in CNS cell cultures.

Liposome-mediated NGF transfection has been shown to increase the expression and secretion of NGF in primary rat septo-hippocampal cell cultures. Here we report that along with increased NGF expression, the activity of choline acetyltransferase, the synthetic enzyme for acetylcholine, is increased by 18% within 2 days, by 41% within 4 days and by 32% within 8 days after NGF gene transfection in septo-hippocampal cell cultures. This result further confirms that biologically active NGF is functionally expressed in septo-hippocampal cells when transfected with cDNA for NGF completed with liposomes.

Animals↗

PubMed1996

Liposome-mediated BDNF cDNA transfer in intact and injured rat brain.

We examined the temporal profile of the expression of brain-derived neurotrophic factor (BDNF) cDNA containing a viral promotor following the injection of liposome cDNA complexes into the intact and traumatically injured rat brain. In situ hybridization and PCR confirmed the presence of injected BDNF cDNA for at least 6 days after injection. A similar profile of BDNF cDNA was observed when it was injected following cortical impact injury. mRNA was also localized around the injection areas. These results suggest that liposome-mediated delivery of neurotrophin cDNA may be a practical gene transfer method for treating traumatic brain injury.

Animals↗

PubMed1996

Inherited and acquired risk factors in colonic neoplasia and modulation by chemopreventive interventions.

The progressively abnormal development of epithelial cells prior to tumor development leads to widely differing chemopreventive approaches. The diversity of these approaches has resulted in different assays to measure the activities of the agents. To apply these assays to preclinical studies, we have developed rodent models in which different stages of evolution of colonic neoplasia are expressed. In one model mice carrying a truncated Apc allele with a nonsense mutation in exon 15 have been generated by gene targeting and embryonic stem cell technology (Apc 1638 mice). These mice develop multiple gastrointestinal lesions including adenomas and carcinomas, focal areas of high grade dysplasia (FAD) and polypoid hyperplasias with FADS. The incidence of inherited colonic neoplasms has now been modulated by a chemopreventive regimen. Colonic lesions significantly increased in Apc 1638 mice on a Western-style diet, compared to Apc 1638 mice on AIN-76A diet which has lower fat content and higher calcium and vitamin D. These studies have also been carried out in normal mice, and have demonstrated without any chemical carcinogen that a Western-style diet induced colonic tumorigenesis. Modulation of cell proliferation has also been induced by Western-style diets in other organs including mammary gland, pancreas and prostate. These findings are leading to the development of new preclinical models for evaluating the efficacy of many classes of chemopreventive agents.

Animals↗

PubMed1996

Dibutyl phthalate purged autologous bone marrow transplant in the treatment of leukemia.

It has been proved that di-N-butyl phthalate (DBP) is singular in killing leukemic cells selectively or accelerating the deterioration of residual leukemic cells in long-term marrow culture in vitro. Based on this principle, the DBP-purged autologous bone marrow transplant has been applied to the treatment of a group of 14 patients suffering from acute nonlymphocytic leukemia. After 5-10 days of in vitro co-culture of marrow cells with DBP at a concentration of 50 micrograms/ml, the recovery of total nucleated cells and the amount of CFU-GM were 67.5% and 68.1%, respectively. In all patients, the reconstitution of hematopoiesis was observed after pre-conditioning and transfusion of purged marrow cells. Among these, two patients had a relapse, two patients died from complications of transplant, one patient died from non-leukemic disease, and the others are all alive and free of disease; the mean survival time as calculated recently was 15 months. These preliminary clinical data support that marrow culture in the presence of DBP is a safe and effective measure for treating leukemia in purged autologous bone marrow transplant.

Adolescent↗

PubMed1996

Community surveillance of coronary heart disease in the Atherosclerosis Risk in Communities (ARIC) Study: methods and initial two years' experience.

The community surveillance component of the Atherosclerosis Risk in Communities (ARIC) Study is designed to estimate patterns and trends of coronary heart disease (CHD) incidence, case fatality, and mortality in four U.S. communities. Community surveillance involves ongoing review of death certificates and hospital discharge records to identify CHD events in community residents aged 35-74 years. Interviews with next of kin and questionnaires completed by physicians and medical examiners or coroners were used to collect information on deaths, and review and abstraction of hospital records were used to collect information on possible fatal and nonfatal myocardial infarctions (MIs). Events were classified using standardized criteria. The initial 2-years' experience with case ascertainment and availability of information needed for classification of events is described. Average annual age-adjusted attack rates of definite MI and CHD mortality rates for blacks in two communities and whites in the four communities are presented and compared with rates based on unvalidated hospital discharge data and vital statistics. Age-adjusted rates based on ARIC classification of definite MI were lower than those based on hospital discharge diagnosis code 410 (e.g., 5.60/1000 and 11.50/1000 among Forsyth County white men, respectively). Age-adjusted rates of definite fatal CHD based on ARIC classification were similarly lower than rates based on underlying cause of death code 410; for example, Jackson black men had rates of 2.82/1000 and 4.52/1000 for definite fatal CHD and UCOD 410-414 or 429.2, respectively.

Adult↗

PubMed1996

Differences between respondents and nonrespondents in a multicenter community-based study vary by gender ethnicity. The Atherosclerosis Risk in Communities (ARIC) Study Investigators.

This study provides data on differences between respondents and nonrespondents by gender and ethnicity in a multicenter community-based study that is rarely collected and that may be useful for estimating bias in prevalence estimates in other studies. Demographic, general health, and cardiovascular risk factors were examined in black and white respondents and nonrespondents to the Atherosclerosis Risk in Communities (ARIC) Study, a prospective study investigating cardiovascular risk factors in approximately 16,000 adults that was initiated in 1986 in four U.S. communities. In one of the communities (Jackson, MS) black participants were recruited exclusively; in another (Forsyth County, NC) 12% of the eligible sample were black, whereas the samples in Washington County, MD and the northwestern suburbs of Minneapolis, MN were almost all white. Demographic and health characteristics were collected during a home interview. Subjects who subsequently agreed to complete a clinical examination were defined as respondents, while eligible participants who only took part in the home interview were considered to be nonrespondents. Approximately 75% of age-eligible individuals (45-64 years) in each community completed the home interview. In three of the communities 86-88% of those who took part in the home interview also completed the clinic examination, whereas only 65% did so in Jackson. Among white participants, response rates were similar in men and women and between communities. Among black participants, the response rates were considerably lower, particularly in men. White male respondents reported a higher socioeconomic status, better general health and a lower prevalence of cardiovascular disease and associated risk factors than white male nonrespondents. The difference between white respondents and nonrespondents were greater for men than women despite similar response rates. Among black participants, respondent/nonrespondent difference were usually of smaller magnitude or absent, particularly in women. General health status and recent hospitalization rates were almost identical in black respondents and nonrespondents. Low response rates can bias estimates of prevalence in community-based studies although differences between respondents and nonrespondents tend to exaggerate real differences between respondents and the eligible population sampled. For example, among white males 25% of respondents and 44% of nonrespondents were current smokers, yet the estimated community prevalence of smoking was 31%. In conclusion, difference observed between respondents and nonrespondents were in the expected direction, but were greater for men than women and for whites than blacks.

Bias↗

PubMed1996

Model of mouse mammary gland hyperproliferation and hyperplasia induced by a western-style diet.

Mammary glands of female C57BL/6J mice were analyzed after they were fed a Western-style diet or control AIN-76A diet. The Western-style diet contained several risk factors found in human diets in geographic regions having increased risk for breast cancer: high fat and phosphate and low calcium and vitamin D. After they were fed these diets for 8, 14, and 20 weeks, mice were sacrificed, and mammary glands were removed for morphometric and radioautographic measurements. Although after the animals were fed the Western-style diet for 8 weeks the number of terminal ducts per mouse mammary gland (NTDMG) was similar in the Western-style and control diet groups, after they were fed the Western-style diet for 14 weeks (p < 0.05) and 20 weeks (p < 0.01) the NTDMG significantly increased compared with the control group. Moreover, there was a significant increase (p < 0.01) in the tritiated thymidine labeling index of mammary terminal ductal epithelial cells after 14 and 20 weeks of Western-style diet administration. Thus the Western-style diet induced increased epithelial cell proliferation and increased NTDMG in female mice when fed during young adult growth and development. The findings raise the possibility that the ingestion of a diet with Western-style fat and phosphate content and with low calcium and vitamin D may induce similar changes during the early development of the human mammary gland.

Animals↗