Biomedical subjects

L Y Yang

Publications and source records attributed to L Y Yang.

At least 37 records · Page 2Linked to original sources

PubMed1998

Influence of male-related stimuli on female postejaculatory refractory period in rats.

Female rats "pace" their sexual contacts with the male when tested in situations where they can escape from the male during copulation. The type and quality of vaginocervical stimulation that the females receive during copulation influences their pacing behavior. This study investigated the effect of several male-related stimuli on the female's postejaculatory refractory period (PER). Females were tested in a two-compartment test chamber from which they could escape the male through one of four openings along the bottom of the barrier separating the two compartments. Experiment 1 examined the influence of the seminal plug, the penile cup, and prostate secretions on the female's PER. Results showed that neither the seminal plug, the penile cup, nor prostate secretions contributed to the female's PER. Experiment 2 investigated the relation of pre-ejaculatory intromission frequency, ejaculation duration, and the number of pelvic thrusts during ejaculation to the female's PER. Results indicated that pre-ejaculatory intromission frequency and ejaculation duration but not the number of pelvic thrusts during ejaculation were significantly correlated with the female's PER. In addition, pre-ejaculatory intromission frequency was significantly correlated with ejaculation duration. Partial correlation analysis suggested that pre-ejaculatory intromission frequency affected ejaculation duration which, in turn, influenced the female's PER. This finding was further supported by the evidence that ejaculation duration and the female's PER were significantly shorter in tests in which the male ejaculated on the first or second intromission.

Animals↗

PubMed1998

IgG subclass/IgM ratio and response to therapy in focal segmental glomerulonecrosis.

Focal segmental glomerulosclerosis (FSGS) is relatively steroid resistant and no clinical or histological marker can predict the response to therapy. To investigate the role of serum immunoglobulin subclass/IgM in predicting the response to therapy in FSGS, serum concentrations of total IgG, IgG subclasses, and the ratio of serum IgG subclasses to total IgG (% IgG subclass) were measured in 27 children during the acute nephrotic state. Prednisolone, cyclophosphamide, and Persantine (dipyridamole) were given for 12 weeks. We divided the patients into good responders or poor responders according to clinical response. The clinical and nephrotic status were similar in both groups. Fourteen patients were good responders with higher serum IgGI/IgM than that of non-responders (4.00+/-0.67 vs. 1.61+/-0.20, P<0.05). There was no significant difference in IgG2/IgM between these two groups. These results suggest that higher serum IgGI/IgM ratios may be associated with a better clinical response. These changes may reflect dysregulation of immunoglobulin class switching in patients with FSGS.

Acute Disease↗

PubMed1998

Clinical efficacy of cyclosporin a neoral in the treatment of paediatric lupus nephritis with heavy proteinuria.

Cyclosporin A (CsA) was introduced in recent years for the treatment of lupus nephritis in patients with steroid resistance or in those with severe corticosteroid toxicity. Our previous study on paediatric patients showed that Neoral (a new microemulsion formulation) had better bioavailability than CsA capsules. To evaluate the clinical efficacy of Neoral in children with lupus nephritis compared with conventional therapy, we performed an open randomized study on 40 children, ranging from 9 to 14 yr old, with class III-V lupus nephritis and heavy proteinuria. They were randomly assigned to either Neoral (5 mg/kg/day), administered q.12.h, or prednisolone (2 mg/kg/day) plus cyclophosphamide (2 mg/kg/day) for 1 yr. Both groups showed a significant decrease in proteinuria (Neoral: 4.62 +/- 1.93 to 0.35 +/- 0.29 g/day, P < 0.05; prednisolone plus cyclophosphamide: 4.52 +/- 1.86 to 0.62 +/- 0.21 g/day, P < 0.01). The CH50 haemolytic assay titre decreased after 1 yr of Neoral treatment (26.5 +/- 0.9 to 21.4 +/- 2.2 U/ml, P < 0.05). Serum C3 and anti-double-stranded (ds) DNA antibody levels also fell with Neoral (C3: 86.2 +/- 6.8 to 76.3 +/- 4.5 mg/dl; anti-ds DNA antibodies: 14.1 +/- 3.2 to 8.2 +/- 1.4 IU/ml, P < 0.05). The Neoral group had a significant increase in growth rate over the prednisolone plus cyclophosphamide group (8.2 +/- 1.1 cm/yr vs 2.7 +/- 0.6 cm/yr, P < 0.01) with improvement of growth status. During the study period, patients tolerated Neoral well with no significant changes in renal function, liver function or lipid profile. Our study implies that Neoral appears to be effective in suppressing proteinuria. Neoral should be regarded as being adjunctive therapy, perhaps with a steroid-sparing effect, in paediatric lupus nephritis. However, its long-term use awaits further studies.

Adolescent↗

PubMed1998

Persistent transforming growth factor-beta 1 expression may predict peritoneal fibrosis in CAPD patients with frequent peritonitis occurrence.

The efficiency of continuous ambulatory peritoneal dialysis depends on the permeability of the peritoneal membrane. Peritoneal fibrosis (PF) causes loss of the dialytic function. Several studies have indicated that PF is closely related to the proliferation of peritoneal fibroblasts and the deposition of extracellular matrix. Transforming growth factor-beta 1 (TGF-beta1) plays a major role in stimulating extracellular matrix deposition. Frequent peritonitis occurrence may cause persistent TGF-beta1 mRNA expression. In an attempt to search for a factor related to PF, we designed a longitudinal study to measure TGF-beta1 levels in dialysate and TGF-beta1 mRNA expression in peritoneal mononuclear cells from peritoneal dialysate before onset, once a week during peritonitis, and after peritonitis in high and low peritonitis occurrence (HPO and LPO) patients. Fifteen patients with a LPO rate and 5 patients with a HPO rate were followed up longitudinally. Meanwhile, TGF-beta1 levels and TGF-beta1 mRNA expression were augmented in peritoneal dialysate effluents before, during, and after the episodes of peritonitis. The peritoneal permeability was evaluated by the peritoneal equilibration test. The results revealed that in the LPO group, TGF-beta1 and TGF-beta1 mRNA were detectable at early stages of peritonitis, but the levels decreased rapidly and were undetectable 2 weeks after peritonitis. On the other hand, in the HPO group, TGF-beta1 and TGF-beta1 mRNA persisted for a long time. We could detect TGF-beta1 and TGF-beta1 mRNA in dialysate effluents and peritoneal mononuclear cells even 2, 3, and 4 weeks after episodes of peritonitis. When compared with that of first or second episode of peritonitis, the peritoneal function evaluated with the peritoneal equilibration test was found to obviously deteriorate during the third episode of peritonitis. These findings were confirmed by an in situ hybridization technique to evaluate the relationship between TGF-beta1 mRNA expression and PF from biopsied peritoneal specimens. These findings suggest that the high TGF-beta1 levels in the dialysate are related to an increased expression of TGF-beta1 in the peritoneum. Thus, the persistent TGF-beta1 expression in the peritoneum may serve as a useful parameter in predicting PF in continuous ambulatory peritoneal dialysis patients with frequent peritonitis occurrence.

Adolescent↗

PubMed1997

Fludarabine triphosphate inhibits nucleotide excision repair of cisplatin-induced DNA adducts in vitro.

Fludarabine (9-beta-arabinofuranosyl-2-fluoroadenine-5'-monophosphate) is clinically active against chronic lymphocytic leukemia and low-grade lymphomas. We reported previously that fludarabine nucleoside synergistically enhanced cisplatin (CDDP)-induced cytotoxicity in vitro, and that the synergism was concomitant with inhibition of removal of cellular CDDP-induced DNA interstrand cross-links, which are presumably repaired by homologous recombinational repair. To extend our work, we investigated whether fludarabine inhibits nucleotide excision repair (NER) of CDDP-induced DNA intrastrand adducts. The effect of fludarabine on NER was determined using a cell-free system in which a plasmid containing the DNA adducts served as the substrate for repair enzymes in whole-cell extracts from repair-competent cells. To prevent the cell-bound high mobility group box-containing proteins from interfering with repair, cell extracts were depleted with high mobility group box proteins by immunoprecipitation prior to the assay. Repair synthesis, measured by the incorporation of [(32)P]dATP or [(32)P]dCTP, was inhibited by 50% at 26 or 43 microM fludarabine triphosphate, respectively; the effect was dose dependent and may have resulted from the termination of repair-patch elongation. These results were consistent with those from pulse-chase experiments demonstrating the conversion of nicked circular plasmid to the closed circular form by cell extracts filling the repair gaps. When proliferating cell nuclear antigen-depleted cell extracts were used and aphidicolin was added in the repair assay to arrest NER at the incision/excision stage, 100 microM fludarabine triphosphate inhibited about 55% of the conversion of nicked plasmids from the closed circular damaged plasmid substrate; the inhibition was dose dependent. We conclude that fludarabine triphosphate inhibited NER at the steps of incision and repair synthesis. These results suggest that fludarabine may serve as a potential repair modulator to improve the antitumor efficacies of combination regimens containing agents that induce NER.

Adenosine Monophosphate↗

PubMed1997

Function of intromissions on intromission-return latency of female rats during paced sexual behavior.

The objectives of this study were to examine how multiple intromissions affect the temporal pattern of the female rat's copulatory behavior; in particular, her latency to return to the male following intromission (intromission-return latency, IRL) and if different hormone replacement regimens affect the temporal aspects of female copulatory behavior. Repeated intromissions alone, without ejaculation, often resulted in prolonged IRLs equal to the postej aculatory refractory period (PER). The first prolonged IRL occurred most frequently between the 24th and 44th intromission. The similar pattern of IRLs around the PER and the prolonged IRLs may indicate that the mechanisms mediating the occurrence of the prolonged IRL are similar to those for the PER. One possible function of the prolonged IRLs may be to facilitate the male's ejaculation after the female has received enough vaginocervical stimulation for the induction of the progestational state of pregnancy. Finally, females receiving a single dose of 50 microg estradiol benzoate (EB) followed by an injection of 0.5 mg progesterone (P) 48 h later showed a significantly longer PER than those receiving 3 daily injections of 0.5 microg EB followed by an injection of 0.5 mg P 24 h after the last EB injection.

Animals↗

PubMed1997

Cyclosporin pharmacokinetics following administration of capsules and Neoral in paediatric patients with lupus nephritis.

AIMS: Neoral is a new microemulsion form of cyclosporin. Pharmacokinetic reports in children are scarce. Therefore, we performed a pharmacokinetic study between Cyclosporin A (CsA) capsules and Neoral in paediatric patients with lupus nephritis. METHODS: A single 5 mg kg(-1) dose orally of either CsA capsules or Neoral was given to 10 paediatric patients (serum creatinine < 1.5 mg dl(-1)). CsA whole blood levels were measured for 24 h post-dose by h.p.l.c. RESULTS: Neoral had a higher C(max) and AUC(C(max): 943 +/- 176 ng ml(-1); AUC: 4612 +/- 785 ng ml(-1) h) than those of the CsA capsules (C(max): 697 +/- 187 ng ml(-1); AUC: 3483 +/- 873 ng ml(-1) h; P < 0.05). There was no difference in t(max) and t(1/2,z) between the two groups. CONCLUSIONS: CsA Neoral had improved absorption and bioavailability, which is similar to what is reported in adults. However, interpatient variability still existed. Careful drug monitoring and dose adjustment should be performed during treatment to avoid nephrotoxicity, especially in lupus nephritis.

Adolescent↗

PubMed1997

Fludarabine-mediated repair inhibition of cisplatin-induced DNA lesions in human chronic myelogenous leukemia-blast crisis K562 cells: induction of synergistic cytotoxicity independent of reversal of apoptosis resistance.

We demonstrated previously that the nucleoside of fludarabine (F-ara-A), a clinically effective agent against chronic lymphocytic leukemia and low-grade lymphoma, produces synergistic cytotoxicity against cisplatin-resistant CP2.0 human colon tumor cells when administered in combination with cisplatin. The purpose of this study was 2-fold: (i) to determine whether the synergy occurs in K562 human chronic myelogenous leukemia cells, which, unlike CP2.0 cells, are relatively resistant to drug-induced apoptosis because they express P210(bcr-abl) and (ii) to study the underlying mechanism for the synergy if the enhancement of cytotoxicity occurs in K562 cells. When K562 cells were treated with fludarabine nucleoside and cisplatin as single agents for 4 hr, IC50 values for fludarabine and cisplatin were 3.33 and 2.28 microM, respectively, as measured by a clonogenic survival assay. The simultaneous treatment of K562 cells with the two agents resulted in synergistic cell killing as determined by median-effect analysis. Such synergistic cell killing by combined cisplatin and fludarabine could not be detected in repair-deficient human xeroderma pigmentosum cell lines. Within the range of cytotoxic concentrations, fludarabine (2.5-15 microM) and cisplatin (3-30 microM) as single agents produced no detectable internucleosomal DNA fragmentation as revealed by gel electrophoresis, nor did the combination of the two drugs induce apoptotic DNA degradation. The effects of fludarabine on the repair of cisplatin-induced DNA adducts and interstrand cross-links in K562 cells were analyzed to determine their correlation with the cytotoxic synergy. The interstrand cross-links were measured by the ethidium bromide binding fluorescence assay and quantitative Southern blotting technique. Repair of the intrastrand adducts was detected with whole-cell extracts using a cisplatin-damaged plasmid as the substrate for the in vitro repair assay. Fludarabine at clinically achievable concentrations (1.5-4.5 microM fludarabine nucleoside; 20-100 microM fludarabine triphosphate) inhibited the repair of the DNA lesions induced by cisplatin in a dose-dependent fashion in K562 cells but not in xeroderma pigmentosum cells. Cotreatment with fludarabine preferentially increased the number of interstrand cross-links induced by cisplatin in actively transcribed genes in K562 cells. These data demonstrate the DNA-repair-inhibitory effect of fludarabine and suggest that this effect may contribute to the synergistic cytotoxicity of the fludarabine/cisplatin combination that resulted in decreased clonogenic survival of apoptosis-resistant K562 cells.

Antineoplastic Agents↗

PubMed1997

Persistent transforming growth factor beta 1 expression may predict peritoneal fibrosis in CAPD patients with frequent peritonitis occurrence.

The efficiency of continuous ambulatory peritoneal dialysis (CAPD) depends on the permeability of the peritoneal membrane. Peritoneal fibrosis (PF) causes the loss of dialytic function. Several studies have indicated that PF is closely related to the proliferation of peritoneal fibroblasts and the deposition of extracellular matrix (ECM). Transforming growth factor beta 1 (TGF beta 1) plays a major role in stimulating ECM deposition. Frequent peritonitis occurrence may cause persistent TGF beta 1 mRNA expression. In an attempt to search for a factor related to PF, we designed a longitudinal study to measure TGF beta 1 levels in dialysate and TGF beta 1 mRNA expression in peritoneal mononuclear cells (PMNCs) from peritoneal dialysate before, at the onset of and once a week during peritonitis and after peritonitis in patients with high peritonitis occurrence (HPO) and patients with low peritonitis occurrence (LPO). Fifteen patients with a LPO rate and 5 patients with a HPO rate were followed up longitudinally. Meanwhile, TGF beta 1 levels and TGF beta 1 mRNA expression were augmented in peritoneal dialytic fluid before, during, and after the episodes of peritonitis. Peritoneal permeability was evaluated by the peritoneal equilibration test (PET). The results revealed that in the LPO group, TGF beta 1 and TGF beta 1 mRNA were detectable at early stages of peritonitis, but the levels decreased rapidly and were undetectable 2 weeks after peritonitis. On the other hand, in the HPO group, TGF beta 1 and TGF beta 1 mRNA persisted for a long time. We could detect TGF beta 1 and TGF beta 1 mRNA in dialytic fluid and PMNCs even 2, 3, and 4 weeks after episodes of peritonitis. When compared with that of the first or second episode of peritonitis, peritoneal function evaluated with the PET was found to obviously deteriorate at the third episode of peritonitis. These findings were confirmed by an in situ hybridization technique to evaluate the relationship between TGF beta 1 mRNA expression and PF from biopsied peritoneal specimens. These findings suggest that the high TGF beta 1 levels in the dialysate are related to an increased expression of TGF beta 1 in the peritoneum. Persistent TGF beta 1 expression in the peritoneum may serve as a useful parameter in predicting PF in CAPD patients with frequent peritonitis occurrence.

Adolescent↗

PubMed1996

Relation of intromissions to the female's postejaculatory refractory period in rats.

The objectives of this study were to investigate the temporal aspects of female sexual behavior during single and multiple ejaculatory tests. Females were tested in a two-compartment chamber where they could escape from the male following sexual contacts. In Experiment 1, correlation analysis showed that the number of intromissions received by the female over 3 ejaculatory series was positively correlated with the female's postejaculatory refractory period (PER). In Experiment 2, females receiving 2-4 intromissions before ejaculation had a PER that did not differ from those receiving 5-15 (average 10) intromissions preceding ejaculation. However, if the male ejaculated on the first or second intromission, the female's PER was significantly shorter than the other groups and did not differ from her return latency after an intromission without ejaculation. Females receiving 24-31 intromissions preceding ejaculation exhibited the longest PER. Analysis revealed that the number of intromissions received by females before ejaculation was positively correlated with the female's PER. We concluded that the male's ejaculatory reflex, seminal emission, and postejaculatory behavior alone without at least 2 preceding intromissions were not sufficient to induce a female's PER comparable to that seen after an ejaculation during normal copulation. In addition, the number of intromissions received by the female preceding ejaculation was positively correlated with the female's PER if the range of intromission frequency was large enough.

Animals↗

PubMed1996

Preincubation of Dacron grafts with recombinant tissue factor pathway inhibitor decreases their thrombogenicity in vivo.

BACKGROUND: We have previously shown that preincubation of whole blood clots with recombinants tissue factor pathway inhibitor (rTFPI) attenuates clot-associated procoagulant activity assessed ex vivo. This study was undertaken to determine whether a single local application of rTFPI induces similar attenuation of the procoagulant activity on preclotted Dacron grafts implanted in an artery in vivo. METHODS: Dacron grafts (4 mm x 4 cm long) were preclotted in porcine blood and incubated with either rTFPI (5 mg/ml) or arginine-phosphate buffer for 15 minutes. Grafts were implanted end-to-end in the femoral arteries of 10 pigs, with one rTFPI-treated and one buffer-treated graft implanted in each animal. Animals did not undergo anticoagulation either before or after graft implantation. Radiolabeled porcine fibrinogen was injected intravenously, and the grafts underwent perfusion for 1 hour. A subgroup of animals (n = 7) also had infusion of radiolabeled autologous platelets at the time of administration of radiolabeled fibrinogen. RESULTS: Fibrin(ogen) deposition was decreased in rTFPI-treated grafts by 36% +/- 7% (mean +/- SEM) compared with buffer-treated grafts (p = 0.001). Platelet deposition was also reduced in the rTFPI-treated grafts by 31% +/- 15%, although the reduction did not reach statistical significance (p = 0.10). The extent of rTFPI-mediated attenuation of fibrin(ogen) versus platelet deposition varied independently among animals. CONCLUSIONS: Clot-directed anticoagulant effects of rTFPI appear to be useful for substantially decreasing the thrombogenicity of Dacron grafts immediately after their implantation. Chronic studies to determine whether the decreases in thrombogenicity result in improved long-term graft patency appear warranted.

Animals↗

PubMed1996

Evaluation of CI-973, a platinum analogue, in refractory or relapsed acute leukemia.

The purpose of the study was to define the maximally tolerated dose (MTD), major toxicities, and possible antitumor activity of CI-973 a new platinum analogue, in patients with refractory or relapsed acute leukemia. CI-973 was given as a 5-day continuous infusion every 3 to 4 weeks to patients with refractory or relapsed acute leukemia, at doses ranging from 150 mg/m2 to 1350 mg/m2 per course. Thirty-six patients were treated including 18 patients with acute myelogenous leukemia (AML), four with acute lymphocytic leukemia (ALL) and 14 with chronic myelogenous leukemia in blastic phase (CML-BP). Severe gastrointestinal and renal side-effects were the dose-limiting toxicities occurring in four of five patients treated with CI-973 1200 to 1350 mg/m2 per course. At the MTD of 1000 mg/m2 per course, three of 13 patients treated (23%) had moderate to severe nausea and vomiting, three (23%) had moderate diarrhea and one had moderate mucositis. Among 21 patients treated at > or = 1000 mg/m2 (15 AML, 6 CML-BP) no objective complete or partial responses were observed. Twelve of 18 patients (66%) with evaluable marrows on day 14 showed significant suppression of marrow blasts percentage and marrow leukemic infiltrate percentage. Tests for measurement of DNA adduct formation in leukemic cells in vivo after CI-973 therapy, and in vitro following exposure of leukemic cells to CI-973 were developed. This study defined the MTD of CI-973 to be 1000 mg/m2 by continuous infusion over 5 days every 3 to 4 weeks in patients with refractory or relapsed acute leukemia. Gastrointestinal and renal side-effects were dose-limiting. No objective responses were noted in this heavily resistant population. Correlations between CI-973-induced DNA adduct formation and individual patient response to CI-973 will help to define its role in leukemia subsets.

Acute Disease↗

PubMed1996

Contribution of de novo fatty acid synthesis to very low density lipoprotein triacylglycerols: evidence from mass isotopomer distribution analysis of fatty acids synthesized from [2H6]ethanol.

A detailed comparison of the structures of plasma very low density lipoprotein (VLDL) and liver triacylglycerols (TG) (Yang et al. 1995. J. Lipid Res. 36: 125-136) has demonstrated that a minimum of 60% of the secreted TG could have been derived from partial lipolysis and reesterification of stored TG and a maximum of 40% could have been derived from direct secretion of newly made TG. To investigate the processes involved in the transfer of TG to VLDL in vivo, we have determined the distribution of deuterium among the molecular species of the liver-TG and VLDL-TG during the infusion of perdeuterated ethanol along with fructose or glucose and during the provision of either glucose or fructose in the drinking water for 2 weeks. The deuterium labeling (percent excess and percent replacement) of the total fatty acids was determined by GC/MS of the methyl esters while the labeling of the glycerol and the glycerol plus fatty acids of the enantiomeric diacylglycerol moieties of TG was determined by LC/MS with on-line mass spectrometry. Supplementation of the diet for 2 weeks with either glucose and fructose stimulated the synthesis of TG containing new fatty acids and glycerol. The proportion of the newly made to preexisting TG differed in VLDL from that in the liver. The 2H % replacement in glycerol and in total fatty acids was greater in VLDL-TG than in the liver-TG. On the basis of the mass isotopomer distribution analysis it was estimated that a maximum of 30% of the VLDL-TG could have been derived directly from TG that was made de novo and did not equilibrate with the liver-TG stores. The transfer of the stored TG to VLDL was best accounted for by a degradation to 2-monoacylglycerols and resynthesis via the 2-monoacylglycerol pathway with addition of an excess of newly synthesized fatty acids to the resynthesized TG.

Animals↗

PubMed1995

Hypoglycemia following ethanol ingestion in children: report of a case.

Ethanol is present in a large number of pharmaceuticals, cosmetics, detergents and beverages. Without adequate safety measures, the accidental ingestion of such products by children is possible. Ethanol ingestion by children is known to cause various metabolic and neurologic disorders, including hypoglycemia, metabolic acidosis, seizures or coma. In Taiwan, little information has been published regarding the effects of ethanol ingestion by children. This is a report of a 5-year-old boy who became hypoglycemic after ingestion of rice wine. Admitted in a coma, the boy regained consciousness within 30 min after slow administration of 5% dextrose. His initial plasma glucose level was extremely low and hypokalemia was also found. Blood ethanol was 159 mg/dL, 3 h after the alcohol ingestion. He was discharged in good health the following day. Ethanol-induced hypoglycemia may be easily overlooked as symptoms of adrenaline excess are frequently absent. Prompt recognition and treatment with intravenous glucose are essential and may be life-saving. Pediatricians need to be aware of the devastating impact of ethanol. Preventive measures, such as child-proof packaging and parent education, are required if ethanol intoxication in children is to be avoided.

Accidents, Home↗

PubMed1995

Biosynthesis of chylomicron triacylglycerols by rats fed glyceryl or alkyl esters of menhaden oil fatty acids.

We have previously shown great similarity in the distribution of fatty acids in the sn-1 and sn-3 positions of the chylomicron triacylglycerols (TG) from rats fed menhaden oil or its ethyl esters, and have proposed that the acylglycerol products of the phosphatidic acid (PA) pathway (ester feeding) are hydrolyzed to 2-monoacylglycerols (2-MG) prior to reconversion to TG via the 2-MG pathway (oil feeding) and secretion as chylomicrons. As the composition of the sn-2-position would also be retained if the TG were hydrolyzed only to the X-1,2-diacylglycerol (DG) stage before resynthesis, we have now retested the hypothesis by determining the molecular association and reverse isomer content of the sn-1,2- and sn-2,3-DG derived from the chylomicron TG and the PA resulting from the two feedings. The new data demonstrate a better than 90% homology among the molecular species of the PA from the oil and ester feeding, along with the characteristic association of the saturated acids with sn-1- and the unsaturated acids with sn-2-position. Due to increased proportion of unsaturated acids in the sn-1-position of the TG, there was only a 15-20% homology between the PA and the sn-1,2-DG moieties of the chylomicron TG from the oil and ester feeding. A lack of homology was also observed between the PA and free sn-1,2-DG, as well as between the free sn-1,2-DG and the sn-1,2-DG moieties of the chylomicron TG. On the basis of molecular association and the sn-1-/sn-3- reverse isomer content of the chylomicron TG a better than 90% homology was recognized between the chylomicron TG resulting from the oil and ester feeding. It is therefore concluded that hydrolysis to 2-MG followed by reesterification via the 2-MG pathway constitutes the most plausible mechanism for the transfer to chylomicrons of the TG arising from alkyl ester feeding.

Alkylation↗

PubMed1995

Origin of triacylglycerol moiety of plasma very low density lipoproteins in the rat: structural studies.

We have compared the molecular species composition of the glycerolipids of rat liver and rat plasma very low density lipoproteins (VLDL). There were differences in the stereospecific distribution of the fatty acids in the triacylglycerols (TG) of the liver and of VLDL. While chiral and reversed phase chromatography with mass spectrometry (LC/MS) revealed great similarities in positional distribution and molecular association of the fatty acids between the sn-1,2-diacylglycerol (DG) moieties of the VLDL and liver TG, the corresponding sn-2,3-DG were distinctly different. The free hepatic sn-1,2-DG and the sn-1,2-DG moiety contained within hepatic phosphatidic acid showed a maximum 60% homology to the sn-1,2-DG contained within the TG of the liver and of VLDL. By contrast, the smaller pool of hepatic free sn-2,3-DG was nearly identical to the sn-2,3-DG moiety contained in the TG of the liver. These differences between hepatic and VLDL TG indicate that direct transfer of hepatic triacylglycerols is not a major mechanism of VLDL TG formation. On the other hand, the results suggest that stored hepatic TG are largely hydrolyzed to sn-1,2-DG and then reesterified to TG before being secreted as VLDL TG. Although an involvement of 2-monoacylglycerol pathway could not be excluded, it probably plays a minor role in VLDL TG formation. Our data suggest that a minimum of 60% of the VLDL TG could have been derived via hydrolysis to DG and reesterification, and a maximum of 40% could have originated via the conventional phosphatidic acid pathway.

Animals↗

PubMed1995

Arabinosyl-2-fluoroadenine augments cisplatin cytotoxicity and inhibits cisplatin-DNA cross-link repair.

Cytotoxicity was increased significantly when arabinosyl-2-fluoroadenine (F-ara-A) was administered in simultaneous combination with cisplatin (CDDP) to human colon tumor cell lines relatively sensitive (LoVo) or resistant (CP2.0) to CDDP. Because the mechanism of action of F-ara-A indicates that it may be an effective inhibitor of DNA repair, we hypothesized that F-ara-A induces cytotoxic augmentation by suppressing cellular repair in CDDP-damaged DNA lesions. To test this, we compared the repair of CDDP-induced DNA interstrand cross-links in the total genome and in ERCC1 gene-specific sequences of LoVo and CP2.0 cells for treatments with CDDP and CDDP plus F-ara-A. We determined the DNA repair by measuring the rate of removal of the cross-links, using two methods, i.e., an ethidium bromide fluorescence binding assay, which detects the DNA lesion in the total genome, and a method combining denaturation/renaturation neutral agarose gel electrophoresis and Southern hybridization to detect gene-specific lesions. When F-ara-A (15 microM) was coadministered with CDDP (15 micrograms/ml for LoVo cells and 30 micrograms/ml for CP2.0 cells) for 4 hr, the initial cross-link index for the total genome was increased 67% (4.5 versus 2.7 with CDDP alone) in LoVo cells and 93% (2.9 versus 1.5 with CDDP alone) in resistant CP2.0 cells. At 10 hr after the treatment, only 5% of the cross-links had been removed in combination-treated LoVo cells, compared with 40% in CDDP-treated LoVo cells; in CP2.0 cells, F-ara-A inhibited the removal of cross-links from 95% to 45%. Similar results were obtained for ERCC1 gene-specific DNA sequences. These data suggest that F-ara-A enhances the accumulation of CDDP-induced cross-links in LoVo and CP2.0 cells by suppressing the repair of such lesions, thereby enhancing the cytotoxicity of CDDP in combination treatment.

Antineoplastic Agents↗

PubMed1994

Comparison of the effect of hyperinsulinemia on acyl-CoA:cholesterol acyltransferase activity in the liver and intestine of the rat.

UNLABELLED: Recent studies have suggested that cholesteryl ester synthesis plays a critical role in the assembly of VLDL apo B and triacylglycerol in the liver. Chronic hyperinsulinemia is associated with increased TG production and since cholesteryl ester synthesis depends on acyl CoA:cholesterol acyltransferase (ACAT), we investigated the possibility that chronic hyperinsulinemia might increase ACAT activity. We also measured ACAT activity in the intestinal mucosa, where it has been suggested to play a role in induction of diabetes-associated hypercholesterolemia. Chronically hyperinsulinemic rats were produced by injecting insulin (2 weeks, 6U/day). To prevent profound hypoglycemia, these rats were given 10% sucrose in place of drinking water. Acute hyperinsulinemia was produced by injecting a single dose of 0.5 U insulin ip. Chronic hyperinsulinemia led to a significant increase in free cholesterol, cholesteryl esters, triacylglycerols and phospholipids in the whole liver (27%, P < 0.05; 60%, P < 0.05; 70%, P < 0.01; 37%, P < 0.01, respectively) and an increase in hepatic microsomal triacylglycerol (P < 0.05). In contrast, the microsomal lipids of the intestinal mucosa decreased significantly. In chronically hyperinsulinemic rats there was no change in hepatic ACAT, while ACAT in the intestine actually decreased (26%-50%, P < 0.01). CONCLUSIONS: since the effect of chronic hyperinsulinemia on hepatic ACAT did not parallel that seen previously on VLDL secretion, the effect of chronic hyperinsulinemia on VLDL production cannot be explained by its effect on hepatic ACAT. However, chronic hyperinsulinemia was associated with reduced ACAT activity in the intestine and this might result in decreased lipoprotein production in the hyperinsulinemic intestine.

Animals↗