SLC26 anion exchangers of guinea pig pancreatic duct: molecular cloning and functional characterization.

The secretin-stimulated human pancreatic duct secretes HCO(3)(-)-rich fluid essential for normal digestion. Optimal stimulation of pancreatic HCO(3)(-) secretion likely requires coupled activities of the cystic fibrosis transmembrane regulator (CFTR) anion channel and apical SLC26 Cl(-)/HCO(3)(-) exchangers. However, whereas stimulated human and guinea pig pancreatic ducts secrete ∼140 mM HCO(3)(-) or more, mouse and rat ducts secrete ∼40-70 mM HCO(3)(-). Moreover, the axial distribution and physiological roles of SLC26 anion exchangers in pancreatic duct secretory processes remain controversial and may vary among mammalian species. Thus the property of high HCO(3)(-) secretion shared by human and guinea pig pancreatic ducts prompted us to clone from guinea pig pancreatic duct cDNAs encoding Slc26a3, Slc26a6, and Slc26a11 polypeptides. We then functionally characterized these anion transporters in Xenopus oocytes and human embryonic kidney (HEK) 293 cells. In Xenopus oocytes, gpSlc26a3 mediated only Cl(-)/Cl(-) exchange and electroneutral Cl(-)/HCO(3)(-) exchange. gpSlc26a6 in Xenopus oocytes mediated Cl(-)/Cl(-) exchange and bidirectional exchange of Cl(-) for oxalate and sulfate, but Cl(-)/HCO(3)(-) exchange was detected only in HEK 293 cells. gpSlc26a11 in Xenopus oocytes exhibited pH-dependent Cl(-), oxalate, and sulfate transport but no detectable Cl(-)/HCO(3)(-) exchange. The three gpSlc26 anion transporters exhibited distinct pharmacological profiles of (36)Cl(-) influx, including partial sensitivity to CFTR inhibitors Inh-172 and GlyH101, but only Slc26a11 was inhibited by PPQ-102. This first molecular and functional assessment of recombinant SLC26 anion transporters from guinea pig pancreatic duct enhances our understanding of pancreatic HCO(3)(-) secretion in species that share a high HCO(3)(-) secretory output.

Functional coupling of apical Cl-/HCO3- exchange with CFTR in stimulated HCO3- secretion by guinea pig interlobular pancreatic duct.

Pancreatic ductal epithelium produces a HCO(3)(-)-rich fluid. HCO(3)(-) transport across ductal apical membranes has been proposed to be mediated by both SLC26-mediated Cl(-)/HCO(3)(-) exchange and CFTR-mediated HCO(3)(-) conductance, with proportional contributions determined in part by axial changes in gene expression and luminal anion composition. In this study we investigated the characteristics of apical Cl(-)/HCO(3)(-) exchange and its functional interaction with Cftr activity in isolated interlobular ducts of guinea pig pancreas. BCECF-loaded epithelial cells of luminally microperfused ducts were alkalinized by acetate prepulse or by luminal Cl(-) removal in the presence of HCO(3)(-)-CO(2). Intracellular pH recovery upon luminal Cl(-) restoration (nominal Cl(-)/HCO(3)(-) exchange) in cAMP-stimulated ducts was largely inhibited by luminal dihydro-DIDS (H(2)DIDS), accelerated by luminal CFTR inhibitor inh-172 (CFTRinh-172), and was insensitive to elevated bath K(+) concentration. Luminal introduction of CFTRinh-172 into sealed duct lumens containing BCECF-dextran in HCO(3)(-)-free, Cl(-)-rich solution enhanced cAMP-stimulated HCO(3)(-) secretion, as calculated from changes in luminal pH and volume. Luminal Cl(-) removal produced, after a transient small depolarization, sustained cell hyperpolarization of approximately 15 mV consistent with electrogenic Cl(-)/HCO(3)(-) exchange. The hyperpolarization was inhibited by H(2)DIDS and potentiated by CFTRinh-172. Interlobular ducts expressed mRNAs encoding CFTR, Slc26a6, and Slc26a3, as detected by RT-PCR. Thus Cl(-)-dependent apical HCO(3)(-) secretion in pancreatic duct is mediated predominantly by an Slc26a6-like Cl(-)/HCO(3)(-) exchanger and is accelerated by inhibition of CFTR. This study demonstrates functional coupling between Cftr and Slc26a6-like Cl(-)/HCO(3)(-) exchange activity in apical membrane of guinea pig pancreatic interlobular duct.

Cytoprotective effect of ulinastatin on LLC-PK1 cells treated with antimycin A, gentamicin, and cisplatin.

The cytoprotective effect of ulinastatin was studied in LLC-PK1 cells treated with antimycin A, gentamicin, or cisplatin. All of the three agents induced a concentration-dependent increase in the release of lactate dehydrogenase and a decrease in the amount of remaining protein. In the cell injury models treated with 1.5 microM antimycin A, 10 mM gentamicin, and 0.3 mM cisplatin, ulinastatin tended to show a cytoprotective effect at a concentration of 3,000 U/ml and provided a significant protective effect at 10,000 U/ml. LLC-PK1 cells treated with 0.3 mM cisplatin, bovine serum albumin, and alpha1-acid glycoprotein at a concentration of 3.54 mg/ml, which is a comparable protein concentration to that of 10,000 U/ml ulinastatin, showed no protective effect but rather enhanced cell injury. These results suggest that ulinastatin exerts a direct protective effect on LLC-PK1 cells against various renal toxicities.

[Survey of studies on the role of play in child nursing].

In this survey, 103 papers submitted to several nursing societies and published in leading nursing journals are outlined and analyzed in terms of motives for studies, kinds of play enjoyed by the subjects and effects of play on members of ward staff, child patients and their family members. The articles are listed in chronological order so that the changes and trends in the studies in this field will be shown. Studies on play started and developed in the clinical field for better ward management and more effective treatment. However, effects of play on development of child patients gradually became the focus of the studies. In terms of the subjects' level of health, it used to be limited to chronically-hospitalized patients who were not behaviorally limited. The scope of the subjects has widened to include "every health level". It's now purported that every patient regardless of his/her health level should enjoy the benefits of play. Regarding kinds of play, indoor plays were initially predominant, but subsequently outdoor activities became more popular with the playing repertoire increasing. Infantile stage has been and still is most frequently discussed in the field, while other developmental stages are now attracting more attention than before.

Protective action of ulinastatin against cisplatin nephrotoxicity in mice and its effect on the lysosomal fragility.

The development of azotemia after cisplatin injection in mice was inhibited by ulinastatin treatment in a dose-dependent manner. Reduction in creatinine clearance and elevation in fractional excretion of sodium in mice receiving cisplatin was ameliorated by ulinastatin administration. Epithelial necrosis and hyaline cast formation in the proximal tubule were also suppressed. Ulinastatin showed no influence on the kidney platinum level after cisplatin injection. In LLC-PK1 cells, addition of ulinastatin to the incubation medium markedly reduced the release of N-acetyl-beta-D-glucosaminidase, on of the lysosomal enzymes, during hypotonic treatment only when cells were damaged with cisplatin. On the other hand, ulinastatin showed no effect on the elevation of malondialdehyde concentration in the murine kidney cortical slices after the treatment with cisplatin. These results indicate that ulinastatin has a protective effect against cisplatin nephrotoxicity, and its prevention of the increase in lysosomal fragility is a probable mechanism involved in the renal protection.

Protective effect of human ulinastatin against gentamicin-induced acute renal failure in rats.

We investigated the protective effect of human ulinastatin against gentamicin-induced acute renal failure in rats. Gentamicin sulfate was subcutaneously injected at a dose of 200 mg/kg for 5 consecutive days. After 3 days administration of gentamicin, a slight decrease in renal function was observed, as well as granulovascular degeneration in the proximal tubular cells as a change in the renal histology. After 5 days administration of gentamicin, a remarkable increase in plasma concentration of creatinine (from 0.27 +/- 0.02 to 1.17 +/- 0.18 mg/dL) and urea nitrogen (from 17.8 +/- 0.6 to 48.8 +/- 5.1 mg/dL) and a significant decrease in creatinine clearance (from 0.64 +/- 0.08 to 0.20 +/- 0.03 mL.100 g-1.min-1) were observed. In addition, an apparent increase in urinary excretion of N-acetyl-beta-D-glucosaminidase and albumin was detected. In the renal histology, proximal tubular necrosis and desquamation of the epithelial cells in the cortex were observed. Furthermore, hyaline cast formation was frequently observed in the outer stripe of the outer medulla. Ulinastatin at doses of 100,000 or 300,000 U/kg was coadministered intraperitoneally just after each gentamicin injection. Ulinastatin treatment showed a dose-dependent suppression of gentamicin-induced biochemical alterations and histological changes. After 5 days treatment with 300,000 U.kg-1.day-1 of ulinastatin, the magnitude of gentamicin-induced changes in renal function was significantly lessened, by 45-80%. The score for proximal tubular injuries and the rate of hyaline cast formation were also significantly lower in the same group of animals than those in the group treated with gentamicin alone. In the in vitro study, ulinastatin at 10-300 U/mL showed a concentration-dependent suppression on the fragility of the lysosomal membrane isolated from rat kidney cortex during hypotonic treatment. These results indicate that human ulinastatin has a prominent protective effect on gentamicin-induced acute renal failure in rats, and the lysosomal membrane stabilizing effect is possibly involved as a mechanism of this action.

Normal and developmental aspects of masochism: transcultural and clinical implications.

Masochism in all its forms has challenged psychoanalytic theories of development a and treatment since Freud (1905, 1919, 1924). It might appear paradoxical to present a concept of normal masochism while masochism has been considered to be so pathological, but I have proposed this concept (Nakakuki 1982, 1984) as a part of normal mental functioning through my experience and study of "traditional" Japanese culture, which I believe contains the features of normal masochism. It is normal in that it represents Japanese adaptation to their cultural orientation and expectations.

Loop and distal actions of a novel diuretic, M17055.

We investigated the mechanism of action of a novel 'high ceiling' diuretic, M17055, in in vivo clearance studies with anesthetized dogs during water diuresis and in vitro microperfusion studies of isolated rabbit renal tubules. In the clearance study, intravenous infusion of M17055 (1 mg/kg per h) decreased free water clearance and increased urinary excretion of Na+ and Cl- to a greater extent than did a maximum dose of furosemide (30 mg/kg per h). With the maximum dose of furosemide, an additional dose of M17055 or hydrochlorothiazide resulted in additional suppression of free water clearance. These results indicate that M17055 has some additional mechanisms of action in the distal nephron. In isolated rabbit cortical thick ascending limb of Henle's loop, M17055 applied to the lumen decreased the lumen positive transepithelial voltage at concentrations over 10(-6) M and suppressed the lumen-to-bath 36Cl- flux at 10(-5) M. In the connecting tubule, M17055 added to the lumen suppressed lumen negative transepithelial voltage in a concentration-dependent manner in a range from 10(-4) to 10(-3) M. The effect of M17055 on transepithelial voltage was also observed in the distal convoluted tubule and cortical collecting duct. Moreover, 10(-3) M of M17055 in the lumen significantly decreased the lumen-to-bath 22Na+ flux in the cortical collecting duct. From these observations, it appears that M17055 acts not only on the thick ascending limb of Henle's loop but also on the distal segments via inhibition of electrogenic Na+ transport.

Pharmacological properties of the novel highly potent diuretic 7-chloro-2,3-dihydro-1-(2-methylbenzoyl)-4(1H)-quinolinone 4-oxime-O-sulfonic acid potassium salt.

7-Chloro-2,3-dihydro-1-(2-methylbenzoyl)-4(1H)-quinolinone 4-oxime-O-sulfonic acid potassium salt (M17055, CAS 114417-20-8) showed potent diuretic and saluretic effects dose-dependently, in rats (p.o.), mice (p.o.) and dogs (i.v.), at doses of 0.1-100 mg/kg, 0.3-100 mg/kg and 0.01-30 mg/kg, respectively. The efficacy of M17055 for diuresis, natriuresis and chloruresis was much higher than that of hydrochlorothiazide and almost the same as that of furosemide. These results indicate that this compound may be classified as a "high ceiling diuretic". The potencies of M17055 for natriuresis in rats (p.o.), mice (p.o.) and dogs (i.v.) calculated with ED50 values were 38, 34 and 24 times, respectively, more potent than those of furosemide. Urinary excretions of sodium, chloride and potassium increased in parallel with urinary volume with the administration of M17055 or furosemide, whereas an apparent dissociation with urinary calcium and sodium excretion was observed with M17055 alone. In rats, the increase of urinary calcium excretion with M17055 was significantly lower than that with furosemide under comparable conditions of natriuresis. Moreover, in mice, M17055 decreased urinary calcium excretion at doses with low effectiveness. In clearance studies using anesthetized dogs, M17055 suppressed negative free water clearance (CH2O) under saline loaded conditions, and it decreased positive CH2O under water diuretic conditions. These changes in the effects on CH2O induced by M17055 resemble those of loop diuretics. However, M17055 could not shift negative CH2O to positive, while furosemide was able to do so. Moreover, positive CH2O decreased to nearly zero with M17055, while urine remained dilute with furosemide even at 30 mg/kg.(ABSTRACT TRUNCATED AT 250 WORDS)

Mechanisms involved in the effect of M6434 on experimental hemorrhagic shock: I. Effects on myocardial contractility and venous return.

To elucidate the mechanisms of protective effect of M6434 on experimental shock, the authors examined the effects of this compound on the survival time and hemodynamic changes in severely hemorrhagic-shocked dogs. We also examined the effects of M6434 on contractile tension of isolated canine ventricular strips and on venous return in dogs with cardiopulmonary bypass in normal and shock state. Intravenous infusion of M6434 at 10 micrograms/kg/min prolonged survival and maintained mean arterial pressure, cardiac output, and first derivative of left ventricular pressure at higher levels than those in the control group, whereas dopamine (10 micrograms/kg/min) did not significantly affect survival time and hemodynamic parameters. M6434 did not change contractile tension in electrically stimulated canine ventricular strips. M6434 (20 micrograms/kg/min) increased the venous return of dogs with cardiopulmonary bypass in both shock and normal state. Phenylephrine (20 micrograms/kg/min) slightly increased venous return in normal state, but not during shock. Dopamine had no effect at 20 micrograms/kg/min, but it increased venous return in both states at 50 micrograms/kg/min. These results suggest that M6434 may improve the hemodynamic derangement in severe hemorrhagic shock through decreasing venous blood pooling.

Mechanisms involved in the effect of M6434 on experimental hemorrhagic shock: II. Effects on energy metabolism and organ blood flow.

Effects of M6434 on survival time and hepatic energy metabolism of hemorrhagic-shocked rats were examined. Effects of the compound on rat mitochondrial respiration and regional blood flow in hemorrhagic-shocked rats were also studied to clarify the mechanisms of the antishock effects. Intravenous infusion of M6434 (3 or 10 micrograms/kg/min) prolonged the survival time of hemorrhagic-shocked rats. M6434 at 10 micrograms/kg/min significantly suppressed the decline of adenosine triphosphate contents and energy charge of the liver, shifted the blood flow distribution from skin and skeletal muscles to vital organs such as the liver and the heart, and also increased cardiac output in hemorrhagic-shocked rats. The mitochondrial respiration was unaffected by M6434 in vitro (10(-6)-10(-5) M). These data suggest that mechanisms of the beneficial effect of M6434 in hemorrhagic-shocked rats may not be based on the direct activation of energy metabolism, but rather on the redistribution of organ blood flow as well as an increase in cardiac output.

Cloned hst gene from normal human leukocyte DNA transforms NIH3T3 cells.

The hst gene was originally identified as a transforming gene in DNAs from stomach cancers and a noncancerous portion of stomach mucosa by transfection assays using NIH3T3 cells (1,2). Subsequently, the hst gene obtained directly from leukocyte DNA of a leukemia patient was sequenced (3,4). Here, cosmid clones containing the hst gene were isolated directly from normal human leukocyte DNA and from T361-2nd-1 cells, a secondary transformant of NIH3T3 cells induced by transfection of DNA from a stomach cancer. All clones containing the hst gene from these different sources transformed NIH3T3 cells with similar efficiency. Restriction map of the hst gene from normal leukocyte DNA was identical with that from leukocyte DNA of a leukemia patient, while the hst gene from T361-2nd-1 cells was rearranged at the 168th nucleotide upstream of the TATA box.

[Prophylactic effects of a combined use of a cephem antibiotic, latamoxef, and tobramycin on postoperative infection in obstetrics and gynecology].

To prevent postoperative infection in obstetric gynecology, latamoxef (LMOX) and tobramycin (TOB) were intravenously administered to 81 patients at daily doses of 2 g and 120 mg, respectively, for 5 postoperative days, and the preventive effect of the combined drugs on postoperative infection were evaluated in terms of the clinical effects and safety. 1. The fever index was low as a whole; 3.38 +/- 2.30 degree hours in patients receiving simple panhysterectomy (n = 61), 3.21 +/- 3.84 degree hours in those given cesarean section (n = 12), and 3.53 +/- 2.78 degree hours in those given other surgical procedures (n = 8). 2. There were no abnormalities in hematological findings or liver and kidney functions 14 days after the surgery. PIVKA-II was observed in 3 of 32 patients 14 days after the operation, at a rate of 9.4%. 3. Except mild diarrhea which was observed in 7 of the 81 patients (8.6%) in 4 to 6 days after the operation, no subjective or objective side effects or postoperative complications were observed. The combined use of LMOX and TOB seems useful for preventing postoperative infection in obstetrics and gynecology in terms of its efficacy and safety.

[Antiviral activity of the human amniotic fluid and placenta].

Antiviral activity (3-24 IU/ml) was detected in the amniotic fluid of 21 of 25 pregnant women examined (84%) by conventional interferon assay using human amniotic cells (WISH) and vesicular stomatitis virus. The pregnancies were from 16 to 42 weeks gestation without obvious viral infection or other complications. The antiviral substance in the amniotic fluid seems to be interferon alpha, because it was not inactivated by heat (56 degrees C, 30 min) or acid (pH 2.0, 24 hrs.) and it was inactivated by anti-human interferon alpha neutralizing serum. Neither the incidence nor the titer of the activity in amniotic fluid has any relationship with gestational age, age of the pregnant women or parity. Antiviral activity (4-32 IU/10 mg protein/ml) was also detected in the placenta homogenate of 23 of 25 pregnant women examined (92%). The pregnancies were from 7 to 41 weeks of gestation with or without complication. The titer of antiviral activity was relatively high in the placenta of babies with central nervous system anomalies and it was low in the placenta of intrauterine growth retardation pregnancies, compared with normal pregnancies. The activity in the placenta was not inactivated by heat (56 degrees C, 30 min), acid (pH 2.0, 24 hrs.), anti-human interferon alpha neutralizing anti-serum or anti-human interferon beta neutralizing anti-serum.

[Fundamental and clinical studies of imipenem/cilastatin sodium in the field of obstetrics and gynecology].

Seven patients with gynecologic infections were treated with the new carbapenem class of antibiotic, imipenem/cilastatin sodium (MK-0787/MK-0791) at a dose level of 500 mg/500 mg or 250 mg/250 mg administered intravenously every 12 hours for 5 days. The results obtained were as follows. Clinical effects of MK-0787/MK-0791 were analyzed in 7 patients, including 1 case with pelvic peritonitis, 2 cases with endometritis, adnexitis and pelvic peritonitis, abdominal abscess, vaginal cuff infection and parametritis, and pyometra. Excellent clinical response was seen in 3 cases and good response in 4 cases. No side effect nor abnormal finding in clinical laboratory values was seen in all patients.

[Clinical efficacy of aztreonam in pelvic peritonitis].

Aztreonam (AZT) was evaluated for its clinical efficacy in 6 patients with pelvic peritonitis. In all the cases, AZT was administered by intravenous injection, and the duration of treatment ranged from 4 to 12 days. Daily dose was 2 g in 4 cases, and 4 g in the remaining 2 cases. Surgical treatment was necessary in 2 cases, whereas in another 2 cases, either LMOX or ABPC was administered in addition to AZT. The clinical results was excellent or good in 5 out of 6 cases. Side effects possibly attributed to AZT were diarrhea in 2 cases and slight increase of GOT and GPT in 1 case, although they disappeared promptly afterward.

[Use of cefminox in infections in the field of obstetrics and gynecology].

A total of 15 cases of obstetrical and gynecological infections was treated with cefminox (CMNX, MT-141), a new cephamycin antibiotic, with following results. The subjects consisted of 3 cases of salpingitis, 2 cases of parametritis, 5 cases of endometritis, and 1 case each of puerperal fever, inflammation of the pelvic dead space, Bartholin's pyocele, vulvar abscess and suppurative mastitis. In 2 cases of endometritis, pelveoperitonitis and adnexitis were complicated, respectively. As a rule, CMNX was administered intravenously at a dosage of 1 g each twice a day by drip infusion route. The clinical results were rated as excellent in 8 cases, good in 6 cases and poor in 1 case, with an efficacy rate of 93.3%. No subjective or objective side effects were seen nor any abnormal laboratory test results were found.

[Clinical efficacy of cefpimizole in inflammatory diseases in the field of gynecology].

Clinical study to evaluate the usefulness of cefpimizole (AC-1370) was made in the 6 patients with female genital organ infections. Two grams or 4 g of AC-1370 was administered a day by intravenous drip infusion ranging from 5 to 18 days. Responses were excellent in 2 cases, good in 3 cases and poor in 1 case, parametritis after hysterectomy. The efficacy rate was 83%. Neither general side effect nor abnormal laboratory finding was observed. AC-1370 showed a satisfactory clinical efficacy in treatment of the infections in the field of gynecology, and it has been concluded that AC-1370 will be an useful antibiotic for these infections.

Natural killer cell activity during pregnancy.

Natural killer cell activity was found to be depressed from the early period of pregnancy to the third trimester, as compared to that in the nonpregnant state. Natural killer cell activity in the puerperium tended to be depressed slightly more than during pregnancy. There were no differences in activity between toxemic and normal pregnant women in the same gestational week. A retrospective survey showed a negative correlation between natural killer cell activity during pregnancy and the birth weight of the baby. Although the role of natural killer cells during pregnancy needs to be more clearly elucidated, they may be involved in fetal growth.

[Clinical efficacy of ceftazidime in inflammatory diseases in the field of gynecology].

Ceftazidime ( CAZ ) was evaluated for its clinical efficacy in a total of 12 cases, namely 3 cases of endometritis, 3 cases of intrapelvic infections, 4 cases of adnexitis and 2 cases of external genital infections. In all the cases, CAZ was administered by intravenous drip infusion, and the duration of the treatment ranged from 3 to 22 days. Daily dose was 2 g in 10 cases, and in the remaining 2 cases, daily doses were changed during the course of treatment in the range from 2 to 4 g. The clinical results of CAZ by disease were as follows; excellent in 1 case and good in 2 cases of endometritis, good in all the 3 cases of intrapelvic infections and the 4 cases of adnexitis, and excellent in 1 case and good in 1 case of external genital infections. The overall efficacy rate was 100%, namely, excellent in 2 cases and good in 10 cases of the total of 12 cases. Neither side effects nor abnormal laboratory findings attributable to CAZ were observed in any of the case. From these results, we may conclude that CAZ is a safe antibiotic with satisfactory clinical effects on gynecological infections.