[Unilateral tuberculosis of middle ear in patient previously treated for rhabdomyosarcoma embryonal in the contralateral middle ear--case report]. / Przypadek jednostronnej gruzlicy ucha srodkowego u chorej leczonej rok wczesniej z powodu rhabdomyosarcoma embryonale przeciwstronnego ucha.

Authors present a case of a female patient surgically treated for chronic otitis media (radical middle ear surgery) in which basing on postoperative pathology report a specific inflammatory process (tuberculosis) has been diagnosed. It is pointed out that in this patient there is a history of malignant neoplastic process (rhabdomyosarcoma embryonal) treated previously at the contralateral middle ear.

Sequence of a putative transporter from rabbit kidney related to the Na+/glucose cotransporter gene family.

A 2 kb cDNA, RK-D, was isolated from a rabbit renal library by hybridization with the Na+/glucose cotransporter (SGLT1) cDNA. The mRNA for RK-D is also approximately 2 kb and is found predominantly in kidney. The RK-D cDNA encodes a protein of 597 amino acids related in sequence to the SGLT family of sodium-coupled transporters, all of which are related to the Na+/glucose cotransporter, SGLT1. Because of the high sequence similarity and conservation of 'signature' family features, it is very likely that RK-D encodes a Na(+)-dependent cotransporter.

Sequence of a pyrimidine-selective Na+/nucleoside cotransporter from pig kidney, pkCNT1.

A 3 kb cDNA called pkCNT1, a new member of the Na+/nucleoside cotransporter family, was cloned from pig kidney and sequenced. The sequence of pkCNT1 encodes a 647 amino acid protein that is 84% identical to the sequence of the rat pyrimidine-selective Na+/nucleoside cotransporter, rCNT1. pkCNT1 transports pyrimidines, such as thymidine and uridine, and has a Km for uridine of 9 microM.

Topology of the Na(+)/dicarboxylate cotransporter: the N-terminus and hydrophilic loop 4 are located intracellularly.

The current secondary structure model of the Na(+)/dicarboxylate cotransporter, NaDC-1, contains 11 transmembrane domains. The model is based on hydropathy analysis and the extracellular location of the carboxy terminus, which contains an N-glycosylation site. In this study, the model was further tested using indirect immunofluorescence of COS-7 cells. The Flag epitope tag (DYKDDDDK) was fused to the amino terminus of NaDC-1 (Flag-NaDC-1), and a monoclonal antibody against the Flag epitope was used to determine the location of the N-terminus. Hydrophilic loop 4 of NaDC-1 was identified using polyclonal antibodies raised against a fusion protein containing amino acids 164--233 of NaDC-1. The expression of NaDC-1 and Flag-NaDC-1 in COS-7 cells was confirmed by functional assays of succinate transport and by Western blots of cell surface biotinylated proteins. Immunofluorescent labeling of cells expressing both NaDC-1 and Flag-NaDC-1 required permeabilization of the plasma membranes with digitonin whereas no immunofluorescence was visible in intact cells. The results of this study show that both the N-terminus and hydrophilic loop 4 of NaDC-1 are located intracellularly, which supports the current model of NaDC-1 structure.

Protein kinase C-mediated regulation of the renal Na(+)/dicarboxylate cotransporter, NaDC-1.

The Na(+)/dicarboxylate cotransporter of the renal proximal tubule, NaDC-1, reabsorbs Krebs cycle intermediates, such as succinate and citrate, from the tubular filtrate. Although long-term regulation of this transporter by chronic metabolic acidosis and K(+) deficiency is well documented, there is no information on acute regulation of NaDC-1. In the present study, the transport of succinate in Xenopus oocytes expressing NaDC-1 was inhibited up to 95% by two activators of protein kinase C, phorbol 12-myristate, 13-acetate (PMA) and sn-1, 2-dioctanoylglycerol (DOG). Activation of protein kinase A had no effect on NaDC-1 activity. The inhibition of NaDC-1 transport by PMA was dose-dependent, and could be prevented by incubation of the oocytes with staurosporine. Mutations of the two consensus protein kinase C phosphorylation sites in NaDC-1 did not affect inhibition by PMA. The inhibitory effects of PMA were partially prevented by cytochalasin D, which disrupts microfilaments and endocytosis. PMA treatment was also associated with a decrease of approximately 30% in the amount of NaDC-1 protein found on the plasma membrane. We conclude that the inhibition of NaDC-1 transport activity by PMA occurs by a combination of endocytosis and inhibition of transport activity.

Molecular evidence for two renal Na+/glucose cotransporters.

Previous studies have shown that two kinetically and genetically distinct Na+/glucose cotransporters exist in mammalian kidney. We have recently cloned and sequenced one of the rabbit renal Na+/glucose cotransporters (SGLT1) and have found that it is identical in sequence to the intestinal Na+/glucose cotransporter. Northern blots showed that SGLT1 mRNA was found predominantly in the outer medulla of rabbit kidney. Injection of mRNA from outer medulla and outer cortex into Xenopus oocytes resulted in equal expression of Na(+)-dependent sugar uptake, indicating that the outer cortex sample contained mRNA encoding both SGLT1 and a second Na+/glucose cotransporter. Western blots using antipeptide antibodies against SGLT1 showed that the SGLT1 protein is more abundant in outer medulla than outer cortex. However, brush border membrane vesicles prepared from outer cortex had a greater capacity for Na(+)-dependent glucose transport, indicating the presence of a second transporter in the vesicles from outer cortex. It appears that the cloned renal Na+/glucose cotransporter, SGLT1, is the 'high affinity, low capacity' transporter found predominantly in outer medulla. There is evidence that a second transporter, the 'low affinity, high capacity' transporter, is in outer cortex. Finally, the cDNA and protein sequences of the two renal Na+/glucose cotransporters are predicted to differ by more than 20%.

High-affinity phlorizin binding in Mytilus gill.

The gill of the marine mussel, Mytilus, contains a high affinity, Na-dependent D-glucose transporter capable of accumulating glucose directly from sea water. We examined the ability of the beta-glucoside, phlorizin, to act as a high-affinity ligand of this process in intact gills and isolated brush border membrane vesicles (BBMV). The time course of association of nanomolar [3H]phlorizin to gills and BBMV was slow, with t50 values between 10 and 30 min, and a half-time for dissociation of approx. 30 min. 1 mM D-glucose reduced equilibrium binding of 1 nM phlorizin by 90-95%, indicating that there was little non-specific binding of this ligand to the gill. In addition, there was little, if any, hydrolysis by the gill of phlorizin to its constituents, glucose and phloretin. Phlorizin binding to gills and BBMV was significantly inhibited by the addition of 50 microM concentrations of D-glucose and alpha-methyl-D-glucose, and unaffected by the addition of L-glucose and fructose. Binding to gills and BBMV was reduced by greater than 90% when Na+ was replaced by K+. Replacement of Na+ by Li+ effectively blocked binding to the intact gill, although Li+ did support a limited amount of glucose-specific phlorizin binding in BBMV. The Kd values for glucose-specific phlorizin binding in intact gills and BBMV were 0.5 nM and 6 nM, respectively. We conclude that phlorizin binds with extremely high affinity to the Na-dependent glucose transporter of Mytilus gill, which may be useful in future efforts to isolate and purify the protein(s) involved in integumental glucose transport.

The substrate recognition domain in the Na+/dicarboxylate and Na+/sulfate cotransporters is located in the carboxy-terminal portion of the protein.

The Na+/dicarboxylate cotransporter, NaDC-1, and the Na+/sulfate cotransporter, NaSi-1, share 43% sequence identity, but they exhibit no overlap in substrate specificity. A functional chimera, SiDC-4, was prepared from NaDC-1 and NaSi-1 by homologous recombination and expressed in Xenopus oocytes. SiDC-4 contains putative transmembrane domains 1-4 of NaSi-1 (amino acids 1-139) and putative transmembrane domains 5-11 of NaDC-1 (amino acids 141-593). SiDC-4 retains the substrate specificity of NaDC-1, which suggests that the substrate recognition domain is found in the carboxy-terminal portion of the protein, past amino acid 141. However, residues that affect substrate affinity and inhibition by furosemide and flufenamate are found in the amino terminal third of the protein. The cation binding properties of SiDC-4, including a stimulation of transport by lithium, differed from both parental transporters, suggesting that cation binding is determined by interactions between the amino- and carboxy-terminal portions of the protein. We conclude that the substrate recognition site of NaDC-1 and NaSi-1 is found in the carboxy-terminal portion of the protein, past amino acid 141, but residues in the amino terminus can affect substrate affinity, inhibitor sensitivity, and cation selectivity.

Influence of estradiol and testosterone on cytochrome P-450 and monooxygenase activity in immature brook trout, Salvelinus fontinalis.

Levels of hepatic microsomal cytochrome P-450 were depressed by administration of estradiol-17 beta and were elevated by administration of testosterone in both male and female juvenile brook trout (Salvelinus fontinalis). Treatment-associated changes in the levels of other microsomal electron transfer components in liver did not reflect the changes in cytochrome P-450 content and were also distinct from the changes in these components in kidney. Electrophoretic analysis of hepatic microsomes revealed that estradiol treatment reduced the amounts of several proteins including some heme-staining protein at 56,000 daltons, possibly containing cytochrome P-450. Hepatic microsomal benzo[a]pyrene hydroxylase and the response to 7,8-benzoflavone in vitro were affected little by steroid treatment, and ethoxyresorufin O-deethylase activity could not be detected in any of the samples. Hepatic microsomes metabolized testosterone to a suite of products including 6 beta-hydroxytestosterone (the major metabolite) and 16 beta-hydroxytestosterone, plus as many as eleven unknown metabolites. Estradiol-17 beta treatment depressed the rates of testosterone metabolism and particularly the rates of 6 beta-hydroxylase activity but did not affect 16 beta-hydroxylase activity. Both activities were largely unaffected by testosterone. The results are consistent with the idea that both androgens and estrogens regulate the levels of hepatic cytochrome P-450 in brook trout and that the effect, at least of estradiol-17 beta, involves regulation of forms that function in specific hydroxylation of testosterone. The significance of these effects and whether factors additional to steroids are involved in this regulation of hepatic cytochromes P-450 in fish remain to be established.

Citrate transport by the kidney and intestine.

Citrate is an important metabolite that is transported in the kidney and intestine. Low urinary citrate concentrations, which may be determined in part by transport processes in the kidney, are associated with the development of kidney stones. Citrate is reabsorbed from the tubular filtrate in the renal proximal tubule on a sodium-coupled transporter, the Na+/dicarboxylate cotransporter, with a broad substrate specificity for Krebs cycle intermediates. The same transporter is found on the brush-border membrane of enterocytes. In contrast to the well-characterized apical pathway for citrate, there is relatively little information about citrate transport across the basolateral membrane. Recently, the complementary DNAs coding for the Na+/citrate transporters from the apical membranes of rabbit and human kidney, NaDC-1 and hNaDC-1, have been cloned and sequenced. These transporters belong to a separate gene family that includes the renal Na+/sulfate cotransporter, NaSi-1.

May-Hegglin anomaly and pregnancy.

A hypertensive patients with thrombocytopenia is reported who had two pregnancies complicated by preeclampsia and cesarean deliveries without hemorrhage. During her first pregnancy corticosteroids were given for presumed autoimmune thrombocytopenia. Thereafter she was splenectomised. Ten years later May-Hegglin anomaly and renal failure were diagnosed. One of her children had easy bruising.

Pregnancy in idiopathic aplastic anemia (report of 10 patients).

This paper reports on 6 patients with severe, 2 with moderate and 2 with mild aplastic anemia who had a total of 18 pregnancies after the diagnosis. All four pregnancies that occurred during the active state of severe and moderate aplastic anemias were electively terminated. Two out of 14 pregnancies that occurred during the long-term remission were electively terminated for non-medical reason, two spontaneous abortions occurred and 10 live births were seen. All offspring were healthy at follow-up. During pregnancy the circulating blood cell levels decreased in 1 out of 6 pregnancies in patients who were in remission from mild and moderate aplastic anemias, and in 4 out of 8 pregnancies in patients who were in remission from severe aplastic anemia. In all 5 cases that showed a relapse during pregnancy the remission recurred following the termination of pregnancy. The data presented suggest that aplastic anemia in long-term remission can unpredictably relapse during pregnancy, but its final outcome appears not to be affected by pregnancy. Furthermore, there is no correlation between the pre-pregnancy clinical course and the events during pregnancy. The outcome of pregnancy during the remission of aplastic anemia seems beneficial, and spontaneous delivery should be preferred.

Postpartum hemolytic uremic syndrome following placental abruption.

Hemolytic uremic syndrome associated with pregnancy is a rare condition. Authors report a patient treated with corticosteroids for bronchial asthma who was afflicted by placental abruption at 24 weeks' gestation. The abruption was preceded by developing herpes zoster and by deteriorating respiratory symptoms. The induced labor was followed by anuria, acute renal failure, microangiopathic hemolytic anemia, thrombocytopenia then fever and hypertension. The patient was treated early with plasma infusion, transfusion and hemodialysis. She recovered completely after 7 weeks. This case seems to be unique inasmuch as the hemolytic uremic syndrome was preceded by prodromal illness during pregnancy and was associated with placental abruption.

Pregnancies and offspring in survivors of acute lymphoid leukemia and lymphoma.

Since few data on the reproductive outlook of patients successfully treated for lymphoproliferative disease are available, further experiences on the pregnancy outcome and offspring follow-up of 12 women treated for acute lymphoid leukemia and 7 women treated for malignant lymphoma are reported. Of the 20 pregnancies of leukemic patients in remission, 14 ended in live births, one in spontaneous abortion, and 5 in elective abortions among which one was performed during relapse. One minor (hip dysplasia) and one major birth defect (Apert syndrome) were seen. The Apert syndrome baby is considered as a new mutation. Of the 9 pregnancies of lymphoma patients 5 ended in normal births and 4 in elective abortions. Neither relapse nor malformation was encountered. One leukemic patient had gestational edema and one lymphoma patient had puerperal thrombophlebitis of lower extremity. The offspring of the above patients appropriately developed and had no complication except for one with cephalhematoma and for another one with epilepsy attributed to birth injury.

Detection and interpretation of lysergic acid diethylamide results by immunoassay screening of urine in various testing groups.

A total of 2259 urine samples were assayed for lysergic acid diethylamide (LSD) using radioimmunoassay (RIA, Coat-a-Count, Diagnostics Products) and a premarket cloned enzyme donor immunoassay (CEDIA, Boehringer Mannheim). Urine samples were obtained from patients admitted to the emergency room, patients in drug rehabilitation programs, and adults and juveniles in criminal probation programs. An overall incidence of positive results was 0.80% for CEDIA (500-pg/mL cutoff) and 0.89% and 0.18% for RIA at cutoffs of 250 and 500 pg/mL, respectively. Of the CEDIA-positive samples, only 17 and 11% were positive by RIA at 250 and 500 pg/mL, respectively, whereas among RIA-positive samples, only 10% of those > 250 pg/mL and only 25% of those > 500 pg/mL were positive by CEDIA. Moreover, only 2 of 25 of samples positive by one of these screening assays were confirmed by gas chromatography-mass spectrometry (GC-MS). It is likely that discrepancies in results between immunoassays are due to differences in antibody specificities used to detect LSD metabolites. In addition, immunoassays may be more sensitive than GC-MS for detecting LSD use as current confirmation assays are targeted towards detection of the parent drug only. The interpretation of results for LSD analysis must be made with knowledge of the limitations for each assay.

Multiple myeloma in pregnancy.

This is a report on pregnancy complicated by multiple myeloma. Severe refractory anemia was present throughout the pregnancy and multiple myeloma was diagnosed in the second trimester. The anemia ceased after delivery but recurred one year later along with other signs of disease progression. The infant remained healthy after a 2-year follow-up.

[Pregnancy in acute lymphoid leukemia]. / Akut lymphoid leukaemiában szenvedök terhességei.

Authors followed up 5 girls, aged 7 to 13 years suffered of acute lymphoid leukemia, who got into complete remission due to induction therapy with multiple cytostatic agents and who became pregnant after the maintenance treatment lasting two and a half years. Of 8 pregnancies of the 5 women 4 pregnancies were terminated by spontaneous deliveries with mature newborns, and 4 ones were interrupted in gestational weeks 7 to 14. Except for one patient who died one year after the termination of pregnancy the other remained in complete remission during and after the pregnancy, and at present their survival times range between 12.5 and 15 years. Among the newborns one had dysplasia coxae congenita of mild grade. Progenies developed healthily in the course of follow-up ranging between 1.5 and 6 years.

[Immune thrombocytopenic purpura and pregnancy: maternal risk factors]. / Immun thrombocytopeniás purpura és terhesség: az anya veszélyeztetettsége.

Authors analyse 75 pregnancies and 51 deliveries, respectively, of 45 mothers with chronic immune thrombocytopenia (ITP) from the point of view of maternal risks. Pregnancies that occurred in active disease were electively terminated. It was found that pregnancy contributed to the clinical manifestation of ITP, and antepartal bleeding took place in 18%, and preeclampsia emerged in 6%. The chronic ITP showed exacerbation during pregnancy in 27.3%, thus causing antepartal bleeding in 4.5% and postpartal bleeding in 15.9%. Postpartal bleeding presented in total 20%, not only in cases with severe ITP, but also in moderate ones. Postpartal blood transfusion was needed in 16%. The postpartal reduction of hemoglobin level determined 2-3 days following delivery was higher than in those ITP patients who had no postpartal haemorrhage than that in the general obstetrical population. This suggests an increased blood loss associated with delivery in patients with ITP. Maternal morbidity of patients with ITP in whom the disease first manifested during pregnancy was higher than that of those patients in whom pregnancy was associated with the remission of ITP. In order to decrease the maternal risks in ITP we advise that pregnancy should occur and be carried during remission.

[Dyserythropoiesis in the fetal liver as an indicator of the intensity of stress caused by abortion]. / Dyserythropoiesis magzati májban, mint a vetélés keltette stress intenzitásának jelzoje.

PIP: The degree of dyserythropoiesis was measured in liver smears and the following values were obtained: 1.6% in fetuses removed during the 1st trimester, 5.9% for those fetuses removed during the 2nd trimester by abortion; 13.5% and 10% in fetuses spontaneously expelled or by induced abortion, respectively; and 30.8% and 22.2% in those fetuses aborted already dead. The degree of hepatic dyserythropoiesis developed in utero increased hypoxia, while it decreased under oxygenized conditions. In babies who died during the neonatal period, significant dyserythropoiesis occurred. All resutls suggest that contractions of the uterus leading to abortion--probably due to uteroplacental hypoxia--cause dyserythropoiesis. Its degree refers to the intensity of the stress. (author's modified)^ieng

[Increased fetal risk of primary venous thrombosis presenting at a young age]. / A fiatalkori primer vénás thrombosis fokozott magzati kockázata.

Based on the hypothesis that the predisposition to thrombosis in women suffering from deep venous thrombosis at young age can disturb also the uteroplacental circulation, the authors retrospectively analyzed the fetal outcome of 333 pregnancies in 101 women with thromboembolic event before 40 years of age and compared it to the fetal outcome of 2943 pregnancies in 1000 randomly selected obstetrical patients without thrombosis. The relative risks of adverse fetal outcomes in thromboembolic women were as follows: 1.85 (95% C.I.: 1.35-2.55) for the spontaneous miscarriage, 3.9 (95% C.I.: 2.20-6.93) for the second-trimester miscarriage, 1.74 (95% C.I.: 1.15-2.64) for the low birth weight, 2.82 (95% C.I.: 1.28-6.30) for the perinatal loss and 7.17 (95% C.I.: 2.64-19.47) for the abruption of placentae. Data obtained suggest that women with deep venous thrombosis at young age should encounter a higher risk of the uteroplacental thrombosis which results in increasing fetal morbidity and mortality during the second and third trimesters of gestation.