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Biomedical subjects

M H Lu

Publications and source records attributed to M H Lu.

At least 37 records · Page 2Linked to original sources

Thyroxine secretion rates during pregnancy in the rat.

Thyroxine secretion rates (TSR) at various stages of pregnancy in rats were measured by the radiothyroxine pool-turnover method. Groups of rats included normal controls (non-pregnant), days 5, 10, 20, and 22 of pregnancy and near term (within 24 hours prior to parturition if past 22 days of pregnancy). Each animal had blood samples taken just prior to injection of 10 microCi L-thyroxine (L-T4)-131I and at 12, 24, 36, and 48 hours afterwards. Determinations of thyroxine iodine (T4-I), volume of distribution (VD), fractional turnover rate (K), and L-T4 pool size were made from these samples. TSR was calculated as the product of (1.54) (T4-I) (VD) (K). An increase in TSR occurred near term, 3.65 micrograms L-T4/day compared to 1.23 micrograms L-T4/day in the controls, was primarily due to an increase in VD from 33.9 ml in the controls to 90.0 ml near term. A rapid increase in TSR to 3.65 micrograms L-T4/day just prior to term was postulated to be due to a stimulatory action of relaxin in the presence of estrogen upon L-T4 utilization. The mechanism of the increase in TSR in pregnant rats is probably due to the effect of several hormonal changes in maternal tissues throughout pregnancy and to adaptation of the pregnant animals to higher metabolic needs of the rapid developing fetuses.

Animals↗

Effect of caloric restriction on aflatoxin B1-induced DNA synthesis, AFB1-DNA binding and cell proliferation in Fischer 344 rats.

Young adult male Fischer rats maintained on a reduced calorie diet (60% of ad libitum food consumption) for 6 weeks showed a decrease in the binding of aflatoxin B1 (AFB1) to hepatic or renal nuclear DNA and a reduction of AFB1-induced hepatocellular damage. Repeated dosing of rats with AFB1 resulted in the inhibition of hepatic and renal DNA synthesis measured by [3H]thymidine incorporation. However, the rate of DNA synthesis was greater in ad libitum (AL) rats than in calorically restricted (CR) animals. Three days after AFB1 dosing, the rate of DNA synthesis had recovered to the control level. Cell cycle analyses measured by a flow cytometric method on kidney cells of both AL and CR rats showed that there were no significant changes in cell populations in the S phase between these two groups of rats. AFB1 inhibited the cell proliferation on an average of 33%. The restoration of the cell proliferation in kidney cells was found on the third day after AFB1 dosing. The rate of the regenerative cell proliferation was found to be slightly greater in AL rats than in CR animals. The AFB1-induced regenerative DNA synthesis in both liver and kidney was retarded by CR.

Aflatoxin B1↗

Cell proliferation by cell cycle analysis in young and old dietary restricted mice.

The effect of dietary restriction (DR) on cell proliferation determined by cell cycle analysis in tissues of young and old mice was investigated. Using the percentage of S-phase cells as an index of cell proliferation, we found that DR inhibited cell proliferation in spleen and thymus in young mice. No significant changes were found in bone marrow and kidney in the ad libitum (AL) or DR mice regardless of age. In old mice, the DR effect was observed in spleen only. When age increased, a parallel decline in cell proliferation was evidenced by a reduced % of S-phase cells. DR produces a greater cell cycle effect in the young mice than in the old mice. The present data suggests that inhibition of cell proliferation by DR may be affected by type of tissue, age, length of DR, and capacity or rate of cell proliferation.

Aging↗

Effects of caloric restriction in animals on cellular function, oncogene expression, and DNA methylation in vitro.

While the life-extending and disease-modulating effects of caloric restriction (CR) are well documented in whole animal studies and in correlative experiments using cells taken from CR animals, very few studies have used cells in culture after their removal from the CR-fed animal. In using this in vivo-->in vitro approach we have attempted to examine the proposition that the effects of CR can be transferred to individual cells by analyzing the cellular functions of proliferation and transformation, the activation of oncogenes, and the methylation of DNA as a function only of diet. Pancreatic acinar cells excised from CR-fed Brown-Norway rats and placed in rich medium showed different responses compared to cells from ad libitum (AL)-fed controls. CR had the effect of slowing growth rate and protecting against spontaneous and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced transformation over 14 passages of cells in culture. At the molecular level, cells from the CR animals showed reduced c-Ha-ras oncogene expression and mutation as well as reduced mutation of the p53 suppressor gene. CR also increased genomic methylation of ras DNA. We conclude that the effects of CR treatment of the animal are transferred to individual cells and note that these responses (decreased proliferation and transformation; depressed oncogene expression and mutation and decreased suppressor gene mutation; and increased oncogene methylation) are cellular and molecular analogs of in vivo weight loss, life extension, and carcinogenesis modulation, which are hallmarks of CR in the whole animal. The fact that these responses are seen generations after the cells are removed from the CR-treated animal indicates that CR causes a permanent predisposition of pancreatic acinar cells to these modulated responses and shows the value of the in vivo-->in vitro protocol in studies that relate diet to cellular and molecular function.

Animals↗

Hepatic nuclear ploidy distribution of dietary-restricted mice.

Hepatic parenchymal cells in most adult mammals are polyploid, with most of the cells in the quiescent or low-proliferation state. Polyploidization has been related to carcinogenesis and aging, and both end points are significantly affected by dietary restriction (DR). Direct measures of hepatic nuclear polyploidization in DR B6C3F1 mice have not been examined. We examined the effect of DR on distributions of nuclear ploidy in both sexes and on different age groups of B6C3F1 mice. Differences between young and old male mice and between old male and female mice were also compared. Hepatic nuclear ploidy values were measured by flow cytometry. The DNA histograms were analyzed for the percentage of nuclei having different classes of DNA content by gating channels between the areas under the peaks of diploid, tetraploid, and octaploid. The results indicate that 1 or 26 months of DR started at 4 months of age did not alter hepatic nuclear ploidy distributions in young and old mice. Our data suggest that in the male mouse, polyploidization is established by 5 months of age for hepatic nuclei and that ploidy classes are affected by sex at 30 months of age. For females, effects in the octaploid nuclei are seen as a result of DR.

Age Factors↗

[Immunohistochemical localization of CEA,EMA and keratin in salivary mucoepidermal carcinoma]

Localizations of CEA,EMA and keration in 19 cases of mucoepidermal carcinomas were investigated using the indirect immunoperoxidase technique.The results showed CEA was negative in normal salivary glands and showed faint reaction in glands near carcinoma tissue.Keratin and EMA were localized in some myoepithelial cells.The positive rates in carcinoma tissue were 78.9%,89.5% and 84.2%,respectively.The positive rates and staining intensity of CEA and EMA in carcinoma tissue gradually decreased with the decline of tumor differentitation,but that of keratin showed no variation.the author consider that CEA and EMA could become good indices in clinically diagnosing mucoepithelial carcinoma and determining tumor differentiation type cell in mucoepidermal carcinoma and have a potential to multiply express the tumor elements of epithelium and/or mesenchyma.

Journal Article↗

Oxidative DNA damage levels in rats fed low-fat, high-fat, or calorie-restricted diets.

Increased fat and caloric content of the diet has been associated with increased mammary tumor incidence. The dietary modulation of cellular redox state may be one mechanism behind this association. We have examined the effects of changes in dietary fat and caloric intake on the levels of 5-hydroxymethyluracil in DNA from rat liver and mammary gland. Female Fischer 344 rats, 40 days old, were maintained on 3% (low-fat), 5% (control), or 20% (high-fat) corn oil diets for 2 weeks. A fourth group of rats had the same daily fat intake as the control group, but total caloric intake was restricted by 40%. As a measure of oxidative DNA damage, 5-hydroxymethyluracil levels were measured in the DNA extracted from liver and mammary gland by gas chromatography-mass spectrometry. 5-Hydroxymethyluracil levels in the liver DNA of the low-fat, high-fat, and calorie-restricted groups were decreased relative to that of control, but the only significant decrease was in the calorie-restricted group (p less than 0.01). In the mammary gland DNA, statistically significant decreases in damage were found in each group relative to control (p less than 0.05). The relationship between fat in the diet and oxidative stress is thus complex. These results show that changes in dietary intake of both fat and calories can modulate oxidative DNA damage levels, and the effect of diet was more clearly evident in the DNA from mammary gland than in DNA from liver.

Animals↗

The vinculin/sarcomeric-alpha-actinin/alpha-actin nexus in cultured cardiac myocytes.

Experiments are described supporting the proposition that the assembly of stress fibers in non-muscle cells and the assembly of myofibrils in cardiac cells share conserved mechanisms. Double staining with a battery of labeled antibodies against membrane-associated proteins, myofibrillar proteins, and stress fiber proteins reveals the following: (a) dissociated, cultured cardiac myocytes reconstitute intercalated discs consisting of adherens junctions (AJs) and desmosomes at sites of cell-cell contact and sub-sarcolemmal adhesion plaques (SAPs) at sites of cell-substrate contact; (b) each AJ or SAP associates proximally with a striated myofibril, and conversely every striated myofibril is capped at either end by an AJ or a SAP; (C) the invariant association between a given myofibril and its SAP is especially prominent at the earliest stages of myofibrillogenesis; nascent myofibrils are capped by oppositely oriented SAPs; (d) the insertion of nascent myofibrils into AJs or into SAPs invariably involves vinculin, alpha-actin, and sarcomeric alpha-actinin (s-alpha-actinin); (e) AJs are positive for A-CAM but negative for talin and integrin; SAPs lack A-CAM but are positive for talin and integrin; (f) in cardiac cells all alpha-actinin-containing structures invariably are positive for the sarcomeric isoform, alpha-actin and related sarcomeric proteins; they lack non-s-alpha-actinin, gamma-actin, and caldesmon; (g) in fibroblasts all alpha-actinin-containing structures are positive for the non-sarcomeric isoform, gamma-actin, and related non-sarcomeric proteins, including caldesmon; and (h) myocytes differ from all other types of adherent cultured cells in that they do not assemble authentic stress fibers; instead they assemble stress fiber-like structures of linearly aligned I-Z-I-like complexes consisting exclusively of sarcomeric proteins.

Actinin↗

Cell cycle analysis in bone marrow and kidney tissues of dietary restricted rats.

The effect of dietary restriction (DR) on the proportion of cells in various phases of the cell cycle as determined by flow cytometry was investigated in the bone marrow and kidney of young and old Fischer 344 rats. Control rats were fed a standard occurrence of numerous age-associated diseases, including cancer, renal diseases and by the control rats starting at 16 weeks of age until killed at 5 or 20 months old. The relative proportion of cells in the various phases of the cell cycle was independent of tissue type, treatment condition and age, consistently showing an order of G1- greater than S- greater than G2M-phase. In old rats DR did not affect cell cycling in bone marrow of either sex, however, it did cause an increase in the percentage of G1-phase cells in the kidney of male rats. Additionally, DR caused a mathematically significant change in the percentage of cells in all phases of the cell cycle in the bone marrow of young male rats but had no effect in young females. The percentage of S-phase cells in both tissues of both sexes decreased in old rats when compared to young rats regardless of treatment conditions, indicating a parallel decline in cell proliferating activity with aging. To summarize, DR produces a greater cell cycle effect in the young male than the old male rats. Proliferative capacity is enhanced when the young male rats are dietary restricted. This may aid in DNA repair mechanisms and/or immune system response.

Aging↗

Differential distribution of subsets of myofibrillar proteins in cardiac nonstriated and striated myofibrils.

Cultured cardiac myocytes were stained with antibodies to sarcomeric alpha-actinin, troponin-I, alpha-actin, myosin heavy chain (MHC), titin, myomesin, C-protein, and vinculin. Attention was focused on the distribution of these proteins with respect to nonstriated myofibrils (NSMFs) and striated myofibrils (SMFs). In NSMFs, alpha-actinin is found as longitudinally aligned, irregular approximately 0.3-microns aggregates. Such aggregates are associated with alpha-actin, troponin-I, and titin. These I-Z-I-like complexes are also found as ectopic patches outside the domain of myofibrils in close apposition to the ventral surface of the cell. MHC is found outside of SMFs in the form of discrete fibrils. The temporal-spatial distribution and accumulation of the MHC-fibrils with respect to the I-Z-I-like complexes varies greatly along the length of the NSMFs. There are numerous instances of I-Z-I-like complexes without associated MHC-fibrils, and also cases of MHC-fibrils located many microns from I-Z-I-like complexes. The transition between the terminal approximately 1.7-microns sarcomere of any given SMF and its distal NSMF-tip is abrupt and is marked by a characteristic narrow alpha-actinin Z-band and vinculin positive adhesion plaque. A titin antibody T20, which localizes to an epitope at the Z-band in SMFs, precisely costains the 0.3-microns alpha-actinin aggregates in ectopic patches and NSMFs. Another titin antibody T1, which in SMFs localizes to an epitope at the A-I junction, typically does not stain ectopic patches and NSMFs. Where detectable, the T1-positive material is adjacent to rather than part of the 0.3-microns alpha-actinin aggregates. Myomesin and C-protein are found only in their characteristic sarcomeric locations (even in just perceptible SMFs). These A-band-associated proteins appear to be absent in ectopic patches and NSMFs.

Actinin↗

Altered hepatic microsomal function and elevated protooncogene expression as residual effects in rats exposed to delta-9-tetrahydrocannabinol.

The microsomal activation of the potent hepatocarcinogen aflatoxin B1 (AFB1) and the expression of selected protooncogenes were investigated in the livers of rats exposed to delta 9-tetrahydrocannabinol (THC). At equimolar levels of cytochrome P-450, the microsome-mediated binding of AFB1 to DNA was significantly lower (56% of the controls) in preparations from drug exposed rats. Hepatic expression of the c-k-ras protooncogene was 3-fold higher in THC exposed animals. These results suggest the possible occurrence of long lasting residual effects in the rats exposed to THC.

Aflatoxin B1↗

Estradiol secretion by the ovarian tissue, in response to hypophyseal stimulation, during ontogenesis of the bullfrog.

Ovaries of tadpoles, froglets, young frogs, and mature frogs of Rana catesbeiana were cut into small pieces. They were incubated for 6 hr in Krebs-Ringer bicarbonate buffer as controls. Another series of ovaries of the same developmental stages were incubated with pituitary extracts in the buffer as experimentals. Media were then analyzed for estradiol secretion by radioimmunoassay. The results showed that estradiol secretion by tadpole ovaries during development was not affected by the addition of pituitary extracts of mature frogs in the media at any stage while those of young and mature frogs with pituitary extracts secreted more estradiol than those without. These findings indicate that tadpole ovaries are unresponsive to pituitary agents to produce estradiol while frog ovaries are dependent on some pituitary hormones to synthesize estradiol. Thus frog ovaries acquire dependence on the pituitary agent only after metamorphosis.

Animals↗

The effect of dietary restriction on myc protooncogene expression in mice: a preliminary study.

The effect of chronic dietary restriction on the expression of the c-myc protooncogene was determined in the livers of a hybrid mouse strain (C57Bl6 x C3H F1 hybrid) at three time points during a 24-h period: 1 h after lights on (1 HALO), 5 h prior to feeding (12 HALO), and 2 h after feeding (19 HALO). In addition, in whole animals studies, changes in core body temperature were monitored. In mice which had been subjected to a chronic diet restriction (60% of the intake of ad libitum controls), c-myc expression was significantly reduced at 1 HALO and 19 HALO compared to corresponding ad libitum animals. In addition, significant differences in c-myc expression were found between time points, in both the ad libitum and restricted groups, suggesting that myc protooncogene expression in the liver may be regulated in a circadian fashion. C-myc expression may correlate with body temperature, suggesting a possible association with metabolic output.

Animals↗

Inhibition of aflatoxin B1 binding to hepatic DNA by dehydroepiandrosterone in vivo.

Dehydroepiandrosterone (DHEA) a naturally occurring steroid, has been reported to inhibit the binding of N-dimethylnitrosamine and 7,12-dimethylbenz[a]anthracene to DNA in vivo and to increase glutathione transferase activity. In this study, we have investigated if DHEA could protect hepatic DNA from damage by the potent hepatocarcinogen aflatoxin B1 (AFB1). Young male Fischer 344 (2-month-old) rats were fed a diet containing 0.8% DHEA for 14 days. Control rats were pair-fed the same diet without DHEA. The rats were then administered a single i.p. dose of [3H]AFB1 in dimethylsulfoxide (0.6 mg/kg body weight; 200 mCi/mmol) and killed after 3 h. Liver weight, mitochondrial, microsomal and cytosolic protein, cytochrome P450 content and glutathione transferase activity increased significantly (P less than 0.001) in DHEA-fed rats; however, the hepatic DNA content was not altered. DHEA feeding increased the total amount of AFB1 bound to hepatic protein but decreased the extent of DNA binding. In in vitro experiments, there was less total binding to DNA and protein by AFB1 when using microsomes from DHEA-fed rats. These results suggest that DHEA inhibits the binding of AFB1 to DNA by modifying the biotransformation of the carcinogen.

Aflatoxin B1↗

In vitro inhibition of estradiol secretion of tadpole ovaries by cyanoketone.

We have demonstrated the presence of delta 5-3 beta-hydroxysteroid dehydrogenase (HSD) activity in tadpole ovaries of Rana catesbeiana. In the present study, we wish to determine whether estradiol (E2) secretion of tadpole ovaries could be influenced by cyanoketone (CK), a specific inhibitor of delta 5-3 beta-HSD. R catesbeina tadpoles at the premetamorphic climax were used, and pooled ovaries were incubated, 30 mg/tube, with CK at dosages of 0, 0.001, 0.01, 0.1, 1, and 10 micrograms/ml of Krebs-Ringer bicarbonate buffer for 6 hr. Media were collected for assay of E2 by radioimmunoassay (RIA). Results showed an inhibition of E2 secretion by CK that was positively correlated with CK dosage, but plateaued at doses of 0.1 microgram/ml and higher. This finding was comparable to that of G.F. Young, H. Kagawa, and Y. Nagahama (1982, Gen. Comp. Endocrinol. 47, 357-360) on adult fish ovaries. However, adult vertebrates depend on gonadotropins to regulate secretion of E2 while tadpoles, being immature, might secrete E2 independently of pituitaries. When the histochemical test for delta 5-3 beta-HSD activity was performed on in vitro CK-treated ovaries, there was a decrease of enzyme activity by CK. The RIA and histochemical findings may contribute to the concept of sex transformation in which a disturbance of steroidogenesis may induce sex reversal from females to males, at least in tadpoles.

Androstenols↗

Teratogenic evaluation of 1,1,3,3-tetrabutylurea in the rat following dermal exposure.

The teratogenic potential of 1,1,3,3-tetrabutylurea (TBU) was evaluated in the Crl:CD(SD)BR rat. Doses of 25, 50 or 100 mg TBU/rat/day in 0.5 ml dimethyl phthalate were applied to the shaven dorsal skin on days 6-15 of gestation, the day on which a sperm-positive vaginal smear was present being designated day 1. The rats were killed 1 day before natural delivery and the foetuses were examined for external development, structure and integrity of internal tissues and organs, and skeletal development. No maternal effects were seen in rats exposed to 25 mg TBU, but skin irritation and a reduction in maternal body-weight gains were seen in rats treated with either 50 or 100 mg/day, the effects being more pronounced in the 100-mg/day group. In the latter group, the number of pregnancies maintained was reduced and the number of resorptions per litter, calculated only on the litters with resorptions, was increased. Foetuses derived from the females treated with 100 mg/day were slightly smaller than the controls but were structurally normal. The outcome of pregnancy was unaltered in rats given either 25 or 50 mg TBU/day. No increase in malformed foetuses was observed in any of the test groups. Under the conditions of this study, TBU was not teratogenic when applied dermally to rats at doses up to 100 mg/day.

Abnormalities, Drug-Induced↗

Teratogenic evaluation of mancozeb in the rat following inhalation exposure.

Crl:CD rats were exposed (whole body) to mancozeb by inhalation at 0, 1, 17, 55, 110, 890, or 1890/500 mg/m3 for 6 hr/day from Day 6 through 15 of gestation (sperm-positive vaginal smear considered Day 1). Dams were killed 1 day prior to natural delivery and fetuses were examined externally, viscerally, and skeletally for any alterations. Maternal toxicity, as evidenced by significantly decreased body weight gain, hindlimb paralysis, general debilitation, and death or termination in extremis, was noted among rats exposed to mancozeb at concentrations of 500 to 1890 mg/m3. Dams from the 55 and 110 mg/m3 groups exhibited decreased body weight gain and hindlimb weakness. There was no maternal toxicity for dams exposed at a concentration of 17 mg/m3. Embryofetal toxicity, as evidenced by a significantly increased incidence of totally resorbed litters, external hemorrhage, and wavy ribs, was noted at concentrations of 55 mg/m3 and above. The embryofetal toxicity occurred only at concentrations toxic to the dam. Among the groups exposed to mancozeb, the incidence of major malformations was not dose related. Hence, under the test conditions of this study, mancozeb was not found to be teratogenic and produced no toxicity unique to the conceptus.

Abnormalities, Drug-Induced↗