Alterations in transporter expression in liver, kidney, and duodenum after targeted disruption of the transcription factor HNF1alpha.

The transcription factor hepatocyte nuclear factor 1alpha (HNF1alpha) is involved in regulation of glucose metabolism and transport, and in the expression of several drug and bile acid metabolizing enzymes. Targeted disruption of the HNF1alpha gene results in decreased Cyp1a2, and Cyp2e1 expression, and increased Cyp4a1 and Cyp7a1 expression, suggesting these enzymes are HNF1alpha target genes. Since hepatic metabolism can be coordinately linked with drug and metabolite transport, this study aims to demonstrate whether HNF1alpha regulates expression of a variety of organic anion and cation transporters through utilization of an HNF1alpha-null mouse model. Expression of 32 transporters, including members of the Oat, Oatp, Oct, Mrp, Mdr, bile acid and sterolin families, was quantified in three different tissues: liver, kidney, and duodenum. The expression of 17 of 32 transporters was altered in liver, 21 of 32 in kidney, and 6 of 32 in duodenum of HNF1alpha-null mice. This includes many novel observations, including marked downregulation of Oats in kidney, as well as upregulation of many Mrp and Mdr family members in all three tissues. These data indicate that disruption of HNF1alpha causes a marked attenuation of several Oat and Oatp uptake transporters in liver and kidney, and increased expression of efflux transporters such as Mdrs and Mrps, thus suggesting that HNF1alpha is a central mediator in regulating hepatic, renal, and intestinal transporters.

Hormonal regulation of renal multidrug resistance-associated proteins 3 and 4 (Mrp3 and Mrp4) in mice.

Multidrug resistance-associated proteins 3 and 4 (Mrp3 and Mrp4) are expressed at much higher levels in female than male kidney. Sex steroids and sex-specific growth hormone (GH) secretion patterns often mediate gender-predominant gene expression. Thus, three models were used to investigate potential endocrine regulation of Mrp3 and Mrp4: (1) gonadectomized (GNX) mice with 17beta-estradiol (E2) or 5alpha-dihydroxytestosterone (DHT) replacement; (2) hypophysectomized (HPX) mice receiving E2, DHT, or simulated male-pattern (MP) or female-pattern (FP) GH secretion; (3) lit/lit mice, which have a spontaneous mutation in the growth-hormone releasing-hormone (GHRH) receptor, with simulated MP- or FP-GH secretion. GNX and HPX decreased Mrp3 mRNA levels compared with intact females. In both respective models E2 administration increased Mrp3 expression in GNX and HPX mice. DHT markedly repressed Mrp3 from GNX+placebo levels, however, this was not observed in the HPX model. In lit/lit mice, Mrp3 expression was lower than in wild-type controls, and MP-GH and FP-GH simulation slightly increased Mrp3 expression. Whereas GNX increased Mrp4 in males to female levels, HPX actually increased Mrp4 expression in both genders +375% and +66%, respectively. In both models DHT markedly repressed Mrp4. Furthermore, Mrp4 was higher in lit/lit than wild-type male mice, and simulation of MP-GH secretion suppressed female-predominant Mrp4 expression. In conclusion, these data indicate that E2 contributes to higher Mrp3 mRNA expression in females, yet a role for androgens in Mrp3 repression cannot be discounted. In contrast, Mrp4 mRNA is higher in females due to repression by both DHT and MP-GH secretion in males.

Tissue distribution and induction of the rat multidrug resistance-associated proteins 5 and 6.

Multidrug resistance-associated proteins (Mrps) are ATP-dependent transporters which transport a wide variety of anionic and cationic compounds. The purpose of this study was to determine the tissue distribution of Mrp5 and 6 in male and female Sprague-Dawley rats in various tissues, and to investigate whether the expression is altered by cholestasis or administration of microsomal enzyme inducers (MEIs). These MEIs activate six different transcriptionally-mediated pathways, and their effects on Mrp5 and Mrp6 expression were determined. The effects of bile-duct ligation, a cholestasis model, on Mrp5 and 6 expression in male rats were quantified. Mrp5 had marked expression in adrenal gland, and moderate expression in cerebral cortex, cerebellum, and stomach. The MEIs polychlorinated biphenyl (PCB)126, phenobarbital, and PCB99 slightly repressed Mrp5, but no single class of receptor agonists induced or repressed Mrp5. Bile-duct ligation tended to increase Mrp5 expression, but was not statistically significant at a 3 day timepoint. Mrp6 expression was highest in intestine, liver, and kidney. Mrp6 was slightly repressed by phenobarbital, dexamethasone, and isoniazid, but no one class of receptor agonists induced or repressed Mrp6, and expression was also unchanged bile-duct ligation. In conclusion, Mrp5 in rats is most highly expressed in the adrenal gland, whereas Mrp6 is mainly expressed in excretory organs (liver, intestine, and kidney), suggesting markedly different functions. Hepatic mRNA levels of Mrp5 or Mrp6 do not seem to be coordinately regulated along with Phase I enzymes via receptor-mediated pathways, and are not part of the hepatoprotective upregulation of basolateral transporters that occurs during cholestasis.

Induction of multidrug resistance protein 3 in rat liver is associated with altered vectorial excretion of acetaminophen metabolites.

Treatment with the microsomal enzyme inducer trans-stilbene oxide (TSO) can decrease biliary excretion of acetaminophen-glucuronide (AA-GLUC) and increase efflux of AA-GLUC into blood. The hepatic canalicular multidrug resistance protein (Mrp) 2 and sinusoidal protein Mrp3 transport AA-GLUC conjugates into bile and blood, respectively. Thus, TSO-induced alterations in the vectorial excretion of AA-GLUC may occur via increased hepatic Mrp3 levels. The goal of this study was to determine whether TSO, diallyl sulfide (DAS), and oltipraz (OLT) treatments can up-regulate Mrp3 protein expression, and whether treatment with DAS and OLT can correspondingly increase hepatovascular efflux of AA metabolites. Rats were administered phenobarbital, TSO, DAS, OLT, or vehicle for 4 days. Interestingly, all of the chemicals increased the plasma concentration and urinary excretion of AA-GLUC and decreased its biliary excretion. In control animals, approximately 77% and 23% of AA-GLUC was excreted into bile or urine, respectively, whereas with inducer-pretreated animals, <32% of AA-GLUC was excreted into bile and >68% was excreted into urine. Correspondingly, all of the compounds increased hepatic Mrp3 mRNA levels by 13- to 37-fold and protein levels by 2- to 6-fold, respectively. In conclusion, these studies correlate increased Mrp3 protein levels in liver with increased hepatovascular excretion of AA-GLUC and suggest that induction of Mrp3 affects the route of drug excretion.

Jaccoud's arthropathy and inflammatory bowel disease.

A patient with longstanding ulcerative colitis developed an arthropathy of the hands and feet characterized by subluxations at the metacarpophalangeal and metatarsophalangeal joints without apparent osseous erosions. This could be a case of Jaccoud's arthropathy associated with inflammatory bowel disease.

Clinical outcomes of patients with suspected pulmonary embolism and low-probability aerosol-perfusion scintigrams.

The clinical outcomes of 183 patients with suspected pulmonary embolism (PE) and a "low probability of PE" interpretation of aerosol-perfusion scintigrams were reviewed. Based in part on the scintigraphic findings, 173 of the patients (94.5%) were not treated with anticoagulants. Over a mean follow-up period of 8.4 months, only one patient (who was from the untreated group) had clinical evidence of PE. These results suggest that a low-probability interpretation of the aerosol-perfusion scintigram is generally reliable and a suitable indicator for clinical decision making in patients suspected of having PE.

Fertility of young mares after long-term anabolic steroid treatment.

The effect of prior treatment with anabolic steroids was studied in 46 three-year-old mares. In the preceding year, these mares had been assigned to 1 of 4 treatment groups and had received the manufacturer's recommended dosage of 1.1 mg of boldenone undecylenate (BU)/kg of body weight, 4.4 mg of boldenone undecylenate (4 BU)/kg, 1.1 mg of nandrolone decanoate (ND)/kg, or 0.04 ml of sesame oil/kg (control, C). Mares had received an injection every 3 weeks for 54 weeks for a total of 19 injections, with the final injection in December. In the following breeding season, fewer (P less than 0.05) mares in all groups previously administered anabolic steroids displayed estrous behavior than did mares in the control group. Duration of estrus was shortened (P less than 0.05) in mares that had received steroids. Abnormal sexual behavior that was observed during steroid treatment continued (P less than 0.05) for up to 6 months after treatment ceased. However, observations of abnormal behavior declined with time (P less than 0.05). All mares in each treatment group ovulated by the end of the trial, and the interval to first ovulation was similar (P greater than 0.05). Ovarian size, follicular development, and conditions of the tubular genitalia was adversely (P less than 0.05) affected in mares in all steroid-treatment groups until approximately the middle of March. After that time, no difference was noted among groups. First-cycle pregnancy rates were 83%, 67%, 50%, and 42% for mares in the untreated, BU, 4 BU, and ND groups, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Tension pneumoperitoneum. A cause of acute aortic occlusion.

Massive accumulation of intraperitoneal air may result in an entity known as tension pneumoperitoneum. The patient usually complains only of abdominal fullness, and the abdomen becomes ballooned, barrel-shaped, and tympanic in all quadrants. Upward displacement of the diaphragm may cause respiratory embarrassment. There may also be diminished venous return due to compression of the inferior vena cava. Abdominal paracentesis may be necessary to improve respiratory exchange before laparotomy. In one patient with a perforated peptic ulcer and tension pneumoperitoneum, an enormous increase in intra-abdominal pressure apparently led to acute aortic occlusion. This particular complication of tension pneumoperitoneum has not previously been reported, to our knowledge. The clinical findings of aortic occlusion were reproduced in a canine model by insufflation of air into the abdomen. An intra-abdominal pressure of 100 mm Hg resulted in loss of the femoral pulse wave measured by an indwelling arterial catheter.

Prediction of solute transport during peritoneal dialysis.

Solute transport, predominantly diffusion, across the peritoneum correlates inversely with molecular weight. Provided that the solute is water soluble, not protein bound, not of unusual density, not ionized, does not have a large hydration shell, and is transported from plasma to dialysate, the peritoneal clearance is predictable over the molecular weight range from 60 to 11,000 daltons. Transport reates that deviate from the predicted can be explained by known physical properties of particular solutes.