Urolithiasis. A study of its frequency

Considering the general impression of an increased number of patients with acute renal colic, the frequencies of roentgenologically verified ureteral and kidney calculi in a Swedish urban district have been studied for the periods 1953-55 and 1968-70. In a material of 986 outpatients (793 men and 193 women) we have proved an increase in incidence for upper urinary tract calculi in men from 2.2 to 3.3 0/00 (p less than 0.001) and in women from 0.5 to 0.8 0/00 (0.01 less than p less than 0.05). For the material as a whole, we have found a 50% increase (from 1.3 to 2.0 0/00; p less than 0.001) of acute urolithiasis between the periods studied. Some implications of the results in connection with primary hyperparathyroidism are discussed.     

Heterotopic pharyngeal hypophyseal adenoma

Juvenile fibroma, carcinoma and sarcoma are the most frequent tumors of the nasopharynx. Ontogenetic tumors that originate from the base of the skull are found very rarely. The case reported deals with a heterotopic pharyngeal pituitary adenoma, that started growing from the epipharynx.     

Glucose and tolbutamide induce apoptosis in pancreatic beta-cells. A process dependent on intracellular Ca2+ concentration

High concentrations of glucose are considered to be toxic for the pancreatic beta-cell. However, the mechanisms underlying beta-cell dysfunction and resulting cell death are not fully characterized. In the present study we have demonstrated that incubation of pancreatic islets and beta-cells from ob/ob mice and Wistar rats with glucose induced a process of apoptotic beta-cell death, as shown by DNA laddering, TdT-mediated dUTP-biotin nick end-labeling (TUNEL) technique, and by using DNA-staining dye HOECHST 33342. The obtained results show that the percentage of apoptotic cells was dependent on glucose concentration, being minimal at 11 mM glucose. At a concentration of 100 microM, aurintricarboxylic acid, an inhibitor of endonuclease activity, almost completely inhibited apoptosis triggered by 17 mM glucose. We have also shown that long term incubation with 100 microM sulfonylurea, tolbutamide, triggered apoptosis in pancreatic beta-cells. The process of beta-cell death induced by high glucose concentration and tolbutamide were Ca2+-dependent, because introduction to the culture medium of 50 microM D-600 or 200 microM diazoxide, which blocked glucose- and tolbutamide-induced [Ca2+]i increase, inhibited apoptosis. Thus, this study shows for the first time that high glucose concentrations and tolbutamide induce apoptosis in pancreatic beta-cells, and that this process is Ca2+-dependent.     

188Re- and 99mTc-MAG3 as prosthetic groups for labeling amines and peptides: approaches with pre- and postconjugate labeling

Either radiolabeled Tc-99m- or Re-188-labeled MAG3-4-nitrophenylester or unlabeled Bz-MAG3-4-nitrophenylester was reacted with amines and peptides to follow a pre- or a postconjugate radiolabeling route, respectively. The model compounds were N'-t-butyloxycarbonyl-1,6-diaminohexane (DH-Boc) and a Lys-protected derivative of the somatostatin analog RC-160 (cyclic D-Phe-Cys-Tyr-D-Trp-Lys-Val-Cys-Trp-NH2). In the case of labeling DH-Boc, both the preconjugate labeling and the postconjugate labeling were found by using analytical HPLC to provide identical radiolabeled compounds regardless whether Re-188 or Tc-99m was used. The results are supported by infrared and mass-spectral data obtained from compounds synthesized using stable rhenium. The 188Re- or 99mTc-MAG3-RC-160 somatostatin analog were synthesized following the preconjugate labeling route and subsequent removal of the protecting group. Biodistributions of 188Re-and 99mTc-MAG3-RC-160 were evaluated in normal and tumor-bearing mice, and were similar to those of radioiodinated 131-RC-160. All radiolabeled analogs of RC-160 were rapidly cleared from the blood and were excreted through the hepatobiliary system with very little normal organ uptake. The tumor uptake (PC-3, human prostate adenocarcinoma) of systemically administered Re-188-MAG3-RC160 was very low, and it reached only 0.28% injected dose/g (%IDg) at 24 h postinjection, similar to what was obtained with I-131-RC-160. Intratumor injections resulted in significant tumor retentions (9.3% ID/g at 24 h).     

Surgical treatment by vitrectomy and autologous serum of 36 idiopathic macular holes

The release of motilin from an isolated preparation of pig duodenum has been studied. There different types of stimuli were applied: electrical nerve stimulation, intraarterially administered peptides, and instillation of test solutions into the lumen of the duodenum. Furthermore extracts of 20 different regions of the pig digestive system have been analyzed for motilin content. Analysis of the extracts only detected significant presence of motilin in the pig duodenum and jejunum (79 +/- 15 and 60 +/- 19 pmol/g). The stimulation experiments showed: (1) a significant noncholinergic depression of motilin release during electrical stimulation of the vagus nerve (nadir at 74 +/- 5% of baseline level; (2) a significant elevation of motilin release in response to intraarterially administered vasoactive intestinal peptide (VIP) (peak at 330 +/- 35% of baseline level), and (3) a significantly elevated motilin release in response to instillation of autologuous bile (peak at 170 +/- 16% of baseline level) and hydrochloric acid (peak at 196 +/- 42% of baseline level) into the duodenal lumen. In conclusion, luminal acidification and bile are important factors in stimulation of motilin release, whereas the vagally stimulated VIP release was insufficient to overcome the general inhibitory effect of vagus stimulation.     

The mucosal attachment at different abutments. An experimental study in dogs

The present experiment was performed to examine if the material used in the abutment part of an implant system influenced the quality of the mucosal barrier that formed following implant installation. 5 beagle dogs were included in the study. The mandibular premolars and the 1st, 2nd and 3rd maxillary premolars were extracted. Three fixtures of the Br?nemark System were installed in each mandibular quadrant (a total of 6 fixtures per animal). Abutment connection was performed after 3 months of healing. In each dog the following types of abutments were used: 2 "control abutments" (c.p. titanium), 2 "ceramic abutments" (highly sintered Al2O3), 1 "gold abutment", and 1 "short titanium abutment". This "short titanium abutment" was provided with an outer structure made of dental porcelain fused to gold. Following abutment connection a plaque control program was initiated and maintained for 6 months. The animals were sacrificed and perfused with a fixative. The mandibles were removed and each implant region was dissected, demineralized in EDTA and embedded in EPON. Semithin sections representing the mesial, distal, buccal and lingual aspects of the peri-implant tissues were produced and subjected to histological examination. The findings from the analysis demonstrated that the material used in the abutment portion of the implant influenced the location and the quality of the attachment that occurred between the periimplant mucosa and the implant. Abutments made of c.p. titanium or ceramic allowed the formation of a mucosal attachment which included one epithelial and one connective tissue portion that were about 2 mm and 1-1.5 mm high, respectively. At sites where abutments made of gold alloy or dental porcelain were used, no proper attachment formed at the abutment level, but the soft tissue margin receded and bone resorption occurred. The abutment fixture junction was hereby occasionally exposed and the mucosal barrier became established to the fixture portion of the implant. It was suggested that the observed differences were the result of varying adhesive properties of the materials studied or by variations in their resistance to corrosion.     

Expression, purification and characterization of the homeodomain of rat ISL-1 protein

Isl-1 is a member of a family of Homeodomains containing proteins that possess an N-terminal pair of zinc binding LIM domains. The Isl-1 gene in rat codes for a protein that binds to the insulin gene enhancer and is also involved in regulation of amylin and proglucagon genes. A DNA sequence coding for 66 amino acid residues containing the C-terminal homeodomain fragment of Isl-1 was expressed as a soluble protein in Escherichia coli. Here, we describe a procedure which allows the rapid native purification of recombinant homeodomain protein fused to an N- terminal tag of six histidines. The purified homeodomain showed DNA- binding activity to its cognate DNA sequence. An enhanced binding activity is observed in the presence of a reducing agent in electrophoretic mobility shift assays. The DNA binding was further characterized by circular dichroism spectroscopy. Addition of DNA to the homeodomain did not change the overall secondary structure content, but the thermal and chemical denaturing profiles were altered. A stabilization of the secondary structure was observed upon DNA binding. The free energy of unfolding at 23 degrees C was 7 kJ mol(-1) in absence of DNA and 29 kJ mol(-1) in the presence of DNA.      

Dopamine-induced endocytosis of Na+,K+-ATPase is initiated by phosphorylation of Ser-18 in the rat alpha subunit and Is responsible for the decreased activity in epithelial cells

Dopamine inhibits Na+,K+-ATPase activity in renal tubule cells. This inhibition is associated with phosphorylation and internalization of the alpha subunit, both events being protein kinase C-dependent. Studies of purified preparations, fusion proteins with site-directed mutagenesis, and heterologous expression systems have identified two major protein kinase C phosphorylation residues (Ser-11 and Ser-18) in the rat alpha1 subunit isoform. To identify the phosphorylation site(s) that mediates endocytosis of the subunit in response to dopamine, we have performed site-directed mutagenesis of these residues in the rat alpha1 subunit and expressed the mutated forms in a renal epithelial cell line. Dopamine inhibited Na+,K+-ATPase activity and increased alpha subunit phosphorylation and clathrin-dependent endocytosis into endosomes in cells expressing the wild type alpha1 subunit or the S11A alpha1 mutant, and both effects were blocked by protein kinase C inhibition. In contrast, dopamine did not elicit any of these effects in cells expressing the S18A alpha1 mutant. While Ser-18 phosphorylation is necessary for endocytosis, it does not affect per se the enzymatic activity: preventing endocytosis with wortmannin or LY294009 blocked the inhibitory effect of dopamine on Na+,K+-ATPase activity, although it did not alter the increased alpha subunit phosphorylation induced by this agonist. We conclude that dopamine-induced inhibition of Na+, K+-ATPase activity in rat renal tubule cells requires endocytosis of the alpha subunit into defined intracellular compartments and that phosphorylation of Ser-18 is essential for this process.     

Glucose decreases Na+,K+-ATPase activity in pancreatic beta-cells. An effect mediated via Ca2+-independent phospholipase A2 and protein kinase C-dependent phosphorylation of the alpha-subunit

In the pancreatic beta-cell, glucose-induced membrane depolarization promotes opening of voltage-gated L-type Ca2+ channels, an increase in cytoplasmic free Ca2+ concentration ([Ca2+]i), and exocytosis of insulin. Inhibition of Na+,K+-ATPase activity by ouabain leads to beta-cell membrane depolarization and Ca2+ influx. Because glucose-induced beta-cell membrane depolarization cannot be attributed solely to closure of ATP-regulated K+ channels, we investigated whether glucose regulates other transport proteins, such as the Na+,K+-ATPase. Glucose inhibited Na+,K+-ATPase activity in single pancreatic islets and intact beta-cells. This effect was reversible and required glucose metabolism. The inhibitory action of glucose was blocked by pretreatment of the islets with a selective inhibitor of a Ca2+-independent phospholipase A2. Arachidonic acid, the hydrolytic product of this phospholipase A2, also inhibited Na+, K+-ATPase activity. This effect, like that of glucose, was blocked by nordihydroguaiaretic acid, a selective inhibitor of the lipooxygenase metabolic pathway, but not by inhibitors of the cyclooxygenase or cytochrome P450-monooxygenase pathways. The lipooxygenase product 12(S)-HETE (12-S-hydroxyeicosatetranoic acid) inhibited Na+,K+-ATPase activity, and this effect, as well as that of glucose, was blocked by bisindolylmaleimide, a specific protein kinase C inhibitor. Moreover, glucose increased the state of alpha-subunit phosphorylation by a protein kinase C-dependent process. These results demonstrate that glucose inhibits Na+, K+-ATPase activity in beta-cells by activating a distinct intracellular signaling network. Inhibition of Na+,K+-ATPase activity may thus be part of the mechanisms whereby glucose promotes membrane depolarization, an increase in [Ca2+]i, and thereby insulin secretion in the pancreatic beta-cell.     

Influence of substrate orientation on the morphologyand orientation of LaNiO3 thin films

LaNiO₃ thin films were successfully prepared by a chemical method from citrate precursors. The LNO precursor solution was spin‐coated onto Si (100) and Si (111) substrates. To obtain epitaxial or highly oriented films, the deposited layers were slowly heated in a gradient thermal field, with a heating rate of 1° min^?1, and annealed at 700°C. The influence of different substrate orientations on the thin film morphology was investigated using atomic force microscopy and X‐ray diffraction analysis. Well‐crystallized films with grains aligned along a certain direction were obtained on both substrates. Films deposited on both substrates were very smooth, but with a different grain size and shape depending on the crystal orientation. Films deposited on Si (100) grew in the (110) direction and had elongated grains, whereas those on Si (111) grew in the (211) direction and had a quasi‐square grain shape.