Hypokalemia-induced downregulation of aquaporin-2 water channel expression in rat kidney medulla and cortex.

Prolonged hypokalemia causes vasopressin-resistant polyuria. We have recently shown that another cause of severe polyuria, chronic lithium therapy, is associated with decreased aquaporin-2 (AQP2) water channel expression (Marples, D., S. Christensen, E.I. Christensen, P.D. Ottosen, and S. Nielsen, 1995. J. Clin. Invest., 95: 1838-1845). Consequently, we studied the effect in rats of 11 days' potassium deprivation on urine production and AQP2 expression and distribution. Membrane fractions were prepared from one kidney, while the contralateral kidney was perfusion-fixed for immunocytochemistry. Immunoblotting and densitometry revealed a decrease in AQP2 levels to 27+/-3.4% of control levels (n=11, P<0.001) in inner medulla, and 34+/-15% of controls (n=5, P<0.05) in cortex. Urine production increased in parallel, from 11+/-1.4 to 30+/-4.4 ml/day (n=11, P<0.01). After return to a potassium-containing diet both urine output and AQP2 labels normalized within 7 d. Immunocytochemistry confirmed decreased AQP2 labeling in principal cells of both inner medullary and cortical collecting ducts. AQP2 labeling was predominantly associated with the apical plasma membrane and intracellular vesicles. Lithium treatment for 24 d caused a more extensive reduction of AQP2 levels, to 4+/-1% of control levels in the inner medulla and 4+/-2% in cortex, in association with severe polyuria. The similar degree of downregulation in medulla and cortex suggests that interstitial tonicity is not the major factor in the regulation of AQP2 expression. Consistent with this furosemide treatment did not alter AQP2 levels. In summary,hypokalemia, like lithium treatment, results in a decrease in AQP2 expression in rat collecting ducts, in parallel with the development of polyuria, and the degree of downregulation is consistent with the level of polyuria induced, supporting the view that there is a causative link.

Day-to-day variation of blood glucose and insulin responses in NIDDM subjects after starch-rich meal.

OBJECTIVE: To study day-to-day variation of postprandial blood glucose and insulin increments in non-insulin-dependent diabetes mellitus (NIDDM) subjects and to analyze intra- and interperson variance of response. RESEARCH DESIGN AND METHODS: Ten NIDDM subjects attending the outpatient clinic at Aarhus Kommunehospital were studied. The subjects ate three meals of 90 g of white bread, with 7 days between tests. RESULTS: Mean +/- SD areas under the blood glucose response curve (above basal) over a 3-h period were 557 +/- 60, 569 +/- 74, and 565 +/- 67 mM x 180 min (NS), and areas under the insulin-response curve were 3350 +/- 448, 2815 +/- 359 and 3551 +/- 679 mU/L x 180 min (NS) on each of the three occasions. The 95% confidence intervals of blood glucose and insulin areas for the test meal repeated three times were 564 +/- 120 mM x 180 min and 3240 +/- 1645 mU/L x 180 min, respectively. Intra- and interperson components of variance were 25 vs. 75% (glucose) and 78 vs. 22% (insulin) of the total variance. The intraperson components of variance included all sources of variation other than between-person variation. There was no significant correlation between blood glucose and insulin response areas. CONCLUSIONS: A valid estimate of the glycemic response in a single patient is obtained after a single meal. Because of the large between-person variation, paired data should preferably be used when comparing glycemic responses to different foods.

Blood glucose and insulin responses to different meals in non-insulin-dependent diabetic subjects of both sexes.

The influence of sex on glucose and insulin responses in patients with non-insulin-dependent diabetes was studied in 12 men and 11 matched women. Two meals of either 100 g white bread or 60 g (raw weight) white rice were given. Blood glucose response areas to white bread (517 vs 509 mmol/L) and to rice (306 vs 353 mmol/L) over a 300-min observation period were similar in females and males, respectively. Insulin responses showed an identical pattern to that of glucose in females and males--35784 vs 28230 pmol/L after white bread and 28044 vs 19464 pmol/L min after rice (NS) over a 300-min observation period, respectively. Within the two study groups, blood glucose-response areas to white bread were significantly higher than those to rice (P less than 0.05), whereas there were no differences in insulin-response areas within or between the two groups. The glycemic index of rice for females (62 +/- 9; mean +/- SE) and males (66 +/- 5) was similar.

Erectile dysfunction in multiple sclerosis.

In a sample of 29 impotent men with multiple sclerosis and erectile problems, penile arterial inflow and venous outflow were within normal limits. In 26 patients, the pudendal evoked potential (PEP) was abnormal, and eight of these also had abnormal bulbocavernous reflex (BCR). Three patients had abnormal PEP and normal BCR, and of these, two had normal and one had abnormal nocturnal erectile activity. The validity of PEP/BCR testing was supported by normal findings in six patients with MS and without erectile problems. Nocturnal erectile activity was normal in 11 patients, of whom nine had abnormal PEP and/or BCR. A high disability score corresponded poorly with both reduced sexual function, insufficient nocturnal erectile activity, and abnormal PEP and/or BCR. Intracavernous injection of papaverine gave erection in 27 patients, the dose needed to create an erection being inversely related to the level of disablement. PEP and BCR testing may be more sensitive in defining neurogenic erectile dysfunction (ED) than nocturnal erectile activity. We considered 26 of the cases to have a neurogenic cause of ED and three to have mainly a psychogenic cause.

Renal growth during pregnancy in insulin-dependent diabetic women. A prospective study of renal volume and clinical variables.

Kidney volume was measured during pregnancy in insulin-dependent diabetic women by an ultrasound technique and prognostic value of these measurements evaluated. A prospective study was performed on 87 pregnant women with insulin-dependent diabetes attending the maternity clinic of Aarhus Kommunehospital. Patients with proliferative retinopathy alone, hydronephrosis, or nephrotic syndrome were excluded. The patients were grouped according to onset and duration of diabetes and to vascular lesions; group I (n = 35, White class B+C), group II (n = 11, White class D0), group III (n = 26, White class D+), and group IV (n = 15, White class F+F/R). The patients visited the hospital every 2 weeks during pregnancy for general obstetric and glycaemic control and blood sampling. The volume of both kidneys was measured by a computerized nephrosonograph during the three terms of pregnancy, the puerperium and 4 months postpartum. The kidney volume increased significantly in all four groups from first to third trimester. In the third trimester the kidney volumes were 375 +/- 68 ml (I), 341 +/- 50 ml (II), 362 +/- 63 ml (III), and 343 +/- 54 ml (IV). The kidney volume in the third trimester was positively correlated with creatinine clearance (r = 0.33, P < 0.01) and inversely correlated with creatinine in serum (r = -0.27, P = < 0.02). Total kidney volume decrease (in percent) defined as the difference of maximal volume and value at 4 months postpartum was inversely correlated to albuminuria in the third trimester (r = -0.25, P < 0.05) and vascular lesions of the patients: (mean +/- SEM) 37 +/- 4% (I), 25 +/- 7% (II), 19 +/- 5% (III), and 11 +/- 7% (IV), P < 0.01. In the puerperium, kidney volume decreased significantly from third trimester in groups I, II, and III, whereas we observed no change in group IV. Six of 15 women in groups II and III with kidney volume < 300 ml and normoalbuminuria in the first trimester developed persistent microalbuminuria after pregnancy (P < 0.02). The renal volume in insulin-dependent diabetic women increases significantly during pregnancy and is inversely related to the vascular lesions of the patients. The decrease in renal volume after pregnancy is related to the albuminuria at the end of pregnancy. Women with longstanding diabetes, White class D (= groups II+III), and kidney volume < 300 ml in the first trimester have a high risk of developing permanent microalbuminuria after pregnancy.

Wireless access to a pharmaceutical database: a demonstrator for data driven Wireless Application Protocol (WAP) applications in medical information processing.

BACKGROUND: The Wireless Application Protocol technology implemented in newer mobile phones has built-in facilities for handling much of the information processing needed in clinical work. OBJECTIVES: To test a practical approach we ported a relational database of the Danish pharmaceutical catalogue to Wireless Application Protocol using open source freeware at all steps. METHODS: We used Apache 1.3 web software on a Linux server. Data containing the Danish pharmaceutical catalogue were imported from an ASCII file into a MySQL 3.22.32 database using a Practical Extraction and Report Language script for easy update of the database. Data were distributed in 35 interrelated tables. Each pharmaceutical brand name was given its own card with links to general information about the drug, active substances, contraindications etc. Access was available through 1) browsing therapeutic groups and 2) searching for a brand name. The database interface was programmed in the server-side scripting language PHP3. RESULTS: A free, open source Wireless Application Protocol gateway to a pharmaceutical catalogue was established to allow dial-in access independent of commercial Wireless Application Protocol service providers. The application was tested on the Nokia 7110 and Ericsson R320s cellular phones. CONCLUSIONS: We have demonstrated that Wireless Application Protocol-based access to a dynamic clinical database can be established using open source freeware. The project opens perspectives for a further integration of Wireless Application Protocol phone functions in clinical information processing: Global System for Mobile communication telephony for bilateral communication, asynchronous unilateral communication via e-mail and Short Message Service, built-in calculator, calendar, personal organizer, phone number catalogue and Dictaphone function via answering machine technology. An independent Wireless Application Protocol gateway may be placed within hospital firewalls, which may be an advantage with respect to security. However, if Wireless Application Protocol phones are to become effective tools for physicians, special attention must be paid to the limitations of the devices. Input tools of Wireless Application Protocol phones should be improved, for instance by increased use of speech control.

[MEDLINE in the age of Internet]. / MEDLINE i Internettets tidsalder.

The use MEDLINE and related resources of literature information has increased significantly with the increased availability and speed of the Internet. However, literature information is presented in many different ways via the Internet and search facilities and web addresses are frequently changed. Many physicians and medical researchers find it difficult to keep track of what is available and many lack skills in effective searching. The present paper presents an overview of the MEDLINE database and related databases available from the National Library of Medicine and searchable through PubMed and Internet Greatfull Med. We discuss strategies for searching MEDLINE and the coverage of MEDLINE related to that of related databases from the NLM and other sources.

[Online publishing in the Internet age]. / Online-publicering i Internettets tidsalder.

The availability of full text medical journal articles is rapidly increasing with the increased availability of the Internet. The potentials of the new technology present researchers, publishers, and librarians with new problems and challenges. Some resources are made available free of charge, whereas others are distributed as parts of large licences negotiated between publishers and consortia of research libraries. How can researchers maintain an overview of the constantly changing resources? How can libraries cope with tasks rapidly redefined by the technology? And how can publishers survive when production and distribution of literature information, including the handling of peer reviewing, might just as well be performed by the researchers themselves or their organisations? The present paper presents some of the resources available and discusses both national and international projects and activities that deal with these questions.

[Clinical aspects of spinal anesthesia administered using 0.5% isobar bupivacaine (Maracine) at the L2/L3 or L4/L5 level]. / Kliniske aspekter af spinalanalgesi anlagt med 0.5% isobar bupivacain (Marcain) på L2/L3 versus L4/L5 niveau.

The effect of employing different interspaces for lumbar puncture during spinal anaesthesia was evaluated in 40 patients receiving 4 ml of 0.5% plain bupivacaine at level L2/L3 or L4/L5. No differences were observed in onset, spread or duration of analgesia. Furthermore, we found that only 80% of the patients, independently of the interspace used, had a cephalad spread to T8 and conclude that spinal anaesthesia using plain bupivacaine is not ideal for supraumbilical surgery.

[Kawasaki disease. Incidence in Denmark during the period 1981-1990]. / Kawasaki's sygdom. Forekomst i Danmark i perioden 1981-1990.

The aim of the study was to estimate the incidence of Kawasaki's disease and to describe the clinical manifestations and the prognosis of the disease in children in Denmark during the period 1981-1990. In a register of all hospital admissions in Denmark, 99 patients were registered as having Kawasaki's disease. Of those, 89 patients fulfilled the clinical diagnostic criteria. The mean incidence of the ten year period was 1.0 per 100,000 children per year. The incidence of the disease decreased in the age group from zero to seven years after which it was rarely observed. Typical laboratory findings were hypersedimentation, leucocytosis and thrombocytosis. Eight children had cardiomegaly and three had electrocardiographic changes. Echocardiography was performed in 66 patients showing coronary artery aneurysms in nine patients (14%) (six boys and three girls). No further cardiac complications were found.

Ultrastructure of three-dimensionally localized distal nephron segments in superficial cortex of the rat kidney.

The ultrastructure of superficial distal nephron segments was analyzed after precise localization of tubule cross sections using computer-assisted three-dimensional reconstructions. Five systems of tubules, each with three interconnected distal tubules, were reconstructed and the lengths of the post macula densa segment of the distal straight tubule (DST), the distal convoluted tubule (DCT), the connecting tubule (CNT), and the initial collecting tubule (ICT) were determined. Each cortical collecting duct (CCD) was in continuity with only one tubule in contact with the renal capsule. In three of the five reconstructions, the two nonsubcapsular tubules fused and had a common connection to the subcapsular tubule. The length, between the macula densa (MD) and the confluence, of subcapsular tubules (2.68 +/- 0.15 mm) significantly exceeded the length of tubules not in contact with the renal capsule (2.05 +/- 0.10 mm). This difference was mainly due to a longer ICT in subcapsular tubules. Subcapsular tubules always contacted the renal capsule in the early DCT and often again in the ICT. Cells in the early DCT showed more microvilli on the luminal surface and more infoldings of basolateral membranes than cells in the late DCT. The ultrastructure of intercalated cells (I cells) varied within a range of different manifestations and the ultrastructural variation of I cells was similar in all the analyzed tubule segments. Connecting tubule cells and principal cells were similar in ultrastructure in all tubule segments and cortical levels analyzed.

Structural adaptation of intercalated cells in rat renal cortex to acute metabolic acidosis and alkalosis.

The structural responses of cells in the distal convoluted, connecting, and collecting tubule to acute acid/base changes were investigated by electron microscopy. Acute metabolic acidosis was induced by administration of ammonium chloride, and acute metabolic alkalosis by potassium or sodium bicarbonate. Morphometric analyses were performed on micrographs of randomly selected distal nephron cells. No structural responses were found in distal convoluted tubule cells, connecting tubule cells, or principal cells but prominent changes were observed in intercalated cells (I cells). Thus, the surface density of the luminal membrane in I cells was significantly higher in acidotic animals and lower in KHCO3 alkalotic animals than in controls. On the contrary, the surface density of the membrane that bounds apical vesicles was higher in KHCO3 alkalotic and lower in acidotic animals than in controls. These results suggest that the luminal membrane is internalized during alkalosis and that the membrane that bounds apical vesicles is transferred to the luminal membrane during acidosis. Since a proton translocating ATPase may be present in the luminal membrane the observations are consistent with the possibility that cortical I cells participate in the maintenance of acid/base homeostasis.

Ultrastructure of distal nephron cells in rat renal cortex.

Distal nephron segments in the rat renal cortex contain distal convoluted tubule cells (DCT cells), connecting tubule cells (CNT cells), intercalated cells (I cells), and principal cells (P cells). The present study was carried out to expand present knowledge on the ultrastructure of these cells. The cells were sampled from superficial cortex and analyzed by electron microscopy. Several morphometric parameters were determined and statistical comparison between cell types was performed. Significant structural differences between the cell types were demonstrated. DCT cells showed the highest volume density of mitochondria whereas the amplification of basolateral membranes was higher in CNT cells than in I and P cells. The surface density of the membrane that bounds intermediate vesicles in the apical cytoplasm was twofold higher in I cells than in the other cell types. The morphological differentiation found in the present study adds to available evidence indicating a functional differentiation between the cell types and provides a reference for structure-function correlations in these cells.

Quantitative morphology of the rat kidney during diabetes mellitus and insulin treatment.

A morphometric study was performed on moderately hyperglycaemic streptozotocin diabetic rats after 10 and 50 days of diabetes, and on groups of rats that, after initial hyperglycaemia for 50 days, were insulin treated for 2 h or for 5, 15 or 38 days. A group of hyperglycaemic diabetic animals were fasted for 18 h. Another group of rats had acute hyperglycaemia induced by intravenous glucose injection. After 10 and 50 days of diabetes, kidney weight was increased by 55 and 93%. Glomerular volume, tubule length, and tubular and interstitial volume increased in diabetic animals compared with controls. After 4 h insulin treatment, the kidney weight was 20% decreased; after 5 days it was 31% decreased. After 38 days the kidney weight was still 26% greater than in controls. In diabetic animals, 18 h fasting induced a 30% decrease in kidney weight. In normal animals, acute hyperglycaemia induced a 22% increase in kidney weight. Volume fractions of most kidney structures remained similar in all groups. However, the glomerular volume fraction was smaller during kidney enlargement, and the tubular volume fraction was larger after induced hyperglycaemia compared with controls. In conclusion, high blood glucose levels in diabetic and normal animals are associated with increased kidney weight. In hyperglycaemic diabetic animals, normalization of blood glucose after insulin treatment or fasting was followed by a decrease in kidney weight.

Effect of spironolactone treatment on the ultrastructure of cortical intercalated cells in the rat kidney.

The ultrastructure of intercalated cells in the rat renal cortex was investigated after 7 days of treatment with the mineralocorticoid inhibitor spironolactone. In untreated and treated animals, two subgroups of IC cells (A-IC and B-IC) could be recognized by electron microscopy, but a substantial fraction of IC cells that corresponded to neither A nor B forms, so-called NonA-NonB, was also found. After spironolactone treatment, qualitative and morphometric analyses showed a decrease in volume density of mitochondria, in cell cross-sectional area, and in boundary length of basolateral membranes in A-IC cells. Among the NonA-NonB-IC cells, some revealed especially abundant mitochondria. Although the frequency of NonA-NonB-IC cells did not change significantly, these mitochondria-rich cells appeared with increased frequency after spironolactone treatment. The ultrastructural changes in A-IC cells might reflect a direct effect of spironolactone on the A-IC cells. However, it is also conceivable that the changes are secondary to functional alterations induced in neighboring segment-specific cells by the experimental conditions.

Tubule-tubule and tubule-arteriole contacts in rat kidney distal nephrons. A morphologic study based on computer-assisted three-dimensional reconstructions.

BACKGROUND: Functional investigations of the tubulo-glomerular feedback mechanism have indicated the existence of a contact between the distal nephron and the macula densa region. The structural justification of such a contact is investigated. EXPERIMENTAL DESIGN: Tubule-tubule and tubule-arteriole contacts were investigated in distal nephrons from normal rat kidneys. Computer-assisted three-dimensional reconstructions of distal nephrons were made from serial sections of renal cortical tissue and selected sections were examined by electron microscopy. RESULTS: In 14 of 15 reconstructed nephrons, the distal convoluted tubule or the connecting tubule approached the macula densa region. A wall-to-wall contact between two tubules corresponding to a three-dimensional distance below 28 microns between the axes of the two tubules was found in only five of the reconstructed tubules. The distal nephron contacts to afferent and efferent arterioles of the same nephron were also examined. The efferent arteriole revealed no consistent contacts but the afferent arteriole contacted the distal convoluted tubule/connecting tubules consistently in all 10 of the superficial nephrons and in 3 of 5 midcortical nephrons. Electron microscopy confirmed a close contact between the distal tubule and the afferent arteriole in superficial nephrons and small nerves were often found at or near the site of contact, but the morphology at the site of contact was not unique. The arteriole contacts were made with late distal convoluted tubules, connecting tubules, or cortical collecting ducts. CONCLUSIONS: In conclusion, the present study shows that tubule-tubule contacts are inconsistent between the macula densa region and the distal nephron but that the tubule-afferent arteriole contact is consistent and close in superficial nephrons. This morphology is compatible with the existence of a feedback mechanism between the superficial distal nephron and the afferent arteriole, apart from the one located at the juxtaglomerular apparatus.

Three-dimensional organization and segmental ultrastructure of rat proximal tubules.

AIMS: To provide a 3-D description and basic morphometric data along the course of subcapsular proximal tubules. METHODS: Proximal convoluted (PCT) and straight (PST) tubules were analyzed in series of up to one thousand 4-micron sections of perfusion-fixed rat renal cortex. Nine proximal tubules were traced through the sections by computer-assisted 3-D reconstruction. Selected sections were reembedded and resectioned for ultrastructural morphometry. RESULTS: Subcapsular PCTs (n = 3) had a total length of 12.0 +/- 0.2 mm (SEM), formed tight clusters and contacted the renal surface from 5 to 10 times. In contrast, mid-cortical tubules (total PT length 11.6 +/- 0.1 mm) and in particular deep cortical tubules (total PT length 12.1 +/- 0.9 mm) extended laterally and intermingled considerably with neighboring tubules. Epithelial height gradually decreased from around 11 microns in the PCT to 7.5 microns in the end of the PST. Brush-border height was around 4 microns in the first PT segment (S1), 2.3 microns in the second segment (S2) and 4.5 microns in the end of the proximal tubule. Tubular wall volume, excluding microvilli, decreased from around 1,450 micron3/microns tubule length in early PCT to 825 micron3/microns in terminal PST. CONCLUSIONS: The results provide, at precisely defined nephron levels, quantitative ultrastructural data relevant to transport physiology.

Lithium-induced structural changes in the cortical distal nephron localized by computer-assisted three-dimensional reconstruction.

Lithium treatment is known to cause tubule dilation in distal nephron segments both in rat and in man. However, due to the heterogeneous cell composition of the distal nephron and the cellular changes following lithium treatment, it has been difficult to identify the structurally changed segments. In this study we have therefore applied computer-assisted reconstruction of cortical distal nephron segments. Tubule dilation was demonstrated in connecting and initial collecting tubules and in the first part of cortical collecting ducts (CCD) whereas it was absent from distal straight and distal convoluted tubules. Principal cells (P cells) in the CCD showed swelling of the cytoplasm, accumulation of actin-like microfilaments, and abnormal arrangements of basolateral membranes. Connecting tubule cells (CNT cells) showed similar but less pronounced changes. Intercalated cells (I cells) showed an accumulation of vesicles in the apical cytoplasm and a reduced luminal surface area. Lesions in P and CNT cells may, at least in part, explain the diabetes insipidus and sodium loss found during lithium treatment. Proton secretion in I cells is probably mediated by an ATPase present in the luminal membrane. The reduction in area of this membrane may explain why lithium-treated animals have a lowered ability to excrete an acid load.