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We describe here a method for determining confidence intervals for a commonly used index of diversity. This approach facilitates the comparison of the genetic population structure of microorganisms isolated from different environments and improves the objective assessment of the discriminatory power of typing techniques.

Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disorder frequently associated with renal failure, hypertension, and other abnormalities. The present study determined whether chronic caffeine intake in an animal model of this disease would affect renal structure and function and blood pressure. Heterozygous male Han:Sprague-Dawley rats with ADPKD and normal littermates were provided with either tap water or solutions of caffeine to drink, starting at 1 month of age. When rats were aged 6 months, glomerular filtration rate (GFR) and mean arterial blood pressure (MAP) were measured under Inactin (Byk Gulden, Konstanz, Germany) anesthesia. Caffeine intake had no effect on GFR or cyst development in rats with PKD. MAP was greater in rats with PKD than normal rats and was increased more by caffeine. The hypertensive effect of chronic caffeine intake could not be ascribed to direct pressor effects of angiotensin II. Based on our finding that caffeine exacerbates hypertension in rats with PKD, it may be prudent for patients with ADPKD to limit coffee consumption to four or fewer cups of caffeinated coffee per day, pending studies of humans.

Background: Few treatments are available to slow the progression to renal failure in autosomal dominant polycystic kidney disease (PKD). In an animal model of PKD, the male heterozygous Han:SPRD rat, intake of a solution of potassium citrate plus citric acid (KCitr) from age one to three months prevented a decline in glomerular filtration rate (GFR). The present study tested whether this beneficial effect is sustained and explored handling of citrate and ammonia in normal and cystic kidneys., Methods: Rats were provided with tap water or citrate solutions to drink, and clearance and survival studies were performed., Results: The GFRs of rats with PKD that consumed KCitr from one month of age were normal at six months of age, while those of their counterparts on water were about one third of normal. Long-term KCitr treatment extended the average life span of rats with PKD from 10 to 17 months. Compared with normal rats, water-drinking rats with PKD had higher plasma [citrate], renal cortical [citrate], and fractional excretion of citrate, and lower rates of renal citrate consumption, ammonia synthesis, and ammonia excretion. Cortical PNH3 was not elevated in cystic kidneys. Intake of Na3 citrate/citric acid solution or K3 citrate solution, but not ammonium citrate/citric acid solution, prevented a decline in GFR in three-month-old rats with PKD., Conclusions: Rats with PKD show abnormal renal handling of citrate and ammonia. Citrate salts that have an alkalinizing effect preserve GFR and extend survival.

The kidney function in a model of autosomal dominant polycystic kidney disease (PKD), the Han:SPRD rat, is dramatically improved by chronic ingestion of a solution of potassium citrate and citric acid (KCitr). This study investigated whether this treatment would also be beneficial in the pcy/pcy mouse, a model of autosomal recessive PKD. Starting at 1 month of age, male CD-1 pcy/pcy and normal CD-1 mice were provided with a solution of 55 mM K(3) citrate/67 mM citric acid or tap water to drink. The pcy/pcy mice on the KCitr solution failed to grow normally and showed elevated plasma urea levels when compared to water-drinking littermates. Growth of normal CD-1 mice was not affected by KCitr intake. The pcy/pcy mice were then provided with a more dilute solution of KCitr to drink: this resulted in greater kidney wet and dry weights and a higher kidney weight/body weight ratio, but no beneficial effects. We conclude that pcy/pcy mice cannot tolerate a high level of KCitr intake and that a lower level is of no benefit. Whether KCitr therapy would be helpful in patients with PKD is still an open question., (Copyright 2000 S. Karger AG, Basel)

Bromfenac sodium (Duract) is a phenylacetic acid-derived nonsteroidal anti-inflammatory agent introduced in the United States in 1997 and withdrawn in 1998. We describe the first case of fulminant hepatic failure associated with this agent treated successfully with liver transplantation. Similarities to hepatotoxicity with related agents is discussed.

Objective: There were two main research questions: First, is there a relationship between rates of child physical abuse, child sexual abuse and child neglect and levels of female and male unemployment, single-parent density and child poverty in the immediately local area; and second, is this relationship different for different categories of abuse and neglect and different categories of deprivation?, Method: Using archival data--registered cases of abuse and neglect and official data on child population, social worker ratio, unemployment rates, single-parent density, means-tested clothing grants and free school meals for children--a multiple correlational analysis was carried out of the 5,551 referrals and 1,450 registered cases of abuse and neglect in Glasgow, Scotland for the years 1991 through to 1993., Results: Substantial correlations were found with all indices of deprivation but particularly physical abuse with rates of male unemployment. Lower and more variable correlations were found with female unemployment rates. Sexual abuse and neglect rates showed a less consistent relationship with the indices of deprivation. In general male unemployment rates alone accounted for two-thirds of the variance in total abuse and neglect rates, other factors adding little or nothing to this., Conclusions: The results demonstrate the importance of selecting small and relatively homogeneous areas for this kind of analysis to achieve ecological validity. Male unemployment rates at this level allow for the ranking of areas in terms of priority need.

We report improved method for white clover (Trifolium repens) transformation using Agrobacterium tumefaciens. High efficiencies of transgenic plant production were achieved using cotyledons of imbibed mature seed. Transgenic plants were recovered routinely from over 50% of treated cotyledons. The bar gene and phosphinothricin selection was shown to be a more effective selection system than nptII (kanamycin selection) or aadA (spectinomycin selection). White clover was transformed with the soybean auxin responsive promoter, GH3, fused to the GUS gene (beta-glucuronidase) to study the involvement of auxin in root development. Analysis of 12 independent transgenic plants showed that the location and pattern of GUS expression was consistent but the levels of expression varied. The level of GH3:GUS expression in untreated plants was enhanced specifically by auxin-treatment but the pattern of expression was not altered. Expression of the GH3:GUS fusion was not enhanced by other phytohormones. A consistent GUS expression pattern was evident in untreated plants presumably in response to endogenous auxin or to differences in auxin sensitivity in various clover tissues. In untreated plants, the pattern of GH3:GUS expression was consistent with physiological responses which are regarded as being auxin-mediated. For the first time it is shown that localised spots of GH3:GUS activity occurred in root cortical tissue opposite the sites where lateral roots subsequently were initiated. Newly formed lateral roots grew towards and through these islands of GH3:GUS expression, implying the importance of auxin in controlling lateral root development. Similarly, it is demonstrated for the first time that gravistimulated roots developed a rapid (within 1 h) induction of GH3:GUS activity in tissues on the non-elongating side of the responding root and this induction occurred concurrently with root curvature. These transgenic plants could be useful tools in determining the physiological and biochemical changes that occur during auxin-mediated responses.

Kidney micropuncture and microdissection studies were carried out on heterozygous 2- to 4-month-old female and male Han:SPRD rats with autosomal dominant polycystic kidney disease (ADPKD) and on normal controls, to determine whether cysts are obstructed. Pressures in proximal tubules and cysts were determined using a servo null device and were recorded before, during, and after intraluminal infusion of an isotonic equilibrium solution at 15 and 50 n1/min. Initial cyst pressures in nine cystic rats averaged 18.5 +/- 5.9 (SD) mm Hg, N = 49, significantly (P < 0.01) higher than in normal proximal tubules in four control rats, 14.3 +/- 1.6 mm Hg, N = 36. Pressures in non-cystic tubules in cystic rats, 16.8 +/- 4.4 mm Hg, N = 25, were not significantly different from pressures in control kidneys or in cysts. When proximal tubules were microinfused at 15 nl/min in control rats, tubule pressure increased by 3.8 +/- 1.2 mm Hg, N = 24. In cysts, the response was highly variable. Twenty out of 33 microinfused cysts (61%) showed responses similar to normal tubules and were considered to be nonobstructed; 13 (39%) showed large pressure increases upon microinfusion, sometimes to values over 100 mm Hg (obstructed cysts). Left kidney inulin clearance (in microliter/min. 100 g body wt) averaged 335 +/- 65 (N = 4) in control rats and 344 +/- 144 (N = 9) in cystic rats; at this early stage of the disease no decline in GFR was seen. Weights of cystic kidneys were twice those of normal animals. Microdissection and scanning electron microscopy revealed the presence of intraluminal casts and debris and constrictions between cysts that would impede fluid flow. We conclude that obstruction is a frequent, early event in PKD and, when present, promotes cyst enlargement. Since many cysts are not obstructed, we suggest that factors other than fixed obstruction initiate cyst formation.

Proanthocyanidins (condensed tannins; PA) purified from the leaves of forage legumes Trifolium arvense, Lotus pedunculatus, Lotus corniculatus, Dorycnium rectum, Coronilla varia, Onobrychis viciifolia, or Hedysarum coronarium, were added to soluble lucerne (Medicago sativa) leaf protein and incubated with strained rumen fluid in vitro. Fractions were collected and frozen immediately. Denatured proteins were fractionated by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE), stained, and relative levels were quantified by densitometry. In the absence of PA the large subunit (LSU) of ribulose bisphosphate carboxylase (EC 4.1.1.39) was susceptible to proteolysis by rumen microflora but the small subunit (SSU) resisted breakdown. PA purified from Onobrychis was added to soluble leaf protein, at PA: protein ratios between 1:1 and 1:20. The rate of proteolysis of LSU was significantly reduced at PA: protein ratios of 1:2 and 1:1 (P < 0.001) and the rate of digestion was reduced by between 3- and 21-fold. In separate experiments PA isolated from the range of species described was added to rumen fluid to give PA: protein ratios of 1:5. The addition of PA significantly reduced the rate of proteolysis of LSU, when compared with PA-free control. There were only small differences between PA from different species. The inhibitory effect of PA may have been due to PA binding to the dietary protein or to the rumen proteases, interfering with the action of proteases on susceptible sites within the substrate.

This paper reports on the production of intergeneric somatic hybrid plants between two sexually incompatible legume species. Medicago sativa (alfalfa, lucerne) leaf protoplasts were inactivated by lethal doses of iodoacetamide. Onobrychis viciifolia (sainfoin) suspension-cell protoplasts were gamma-irradiated at lethal doses. Following electrofusion under optimized conditions about 50,000 viable heterokaryons were produced in each test. The fusion products were cultured with the help of alfalfa nurse protoplasts. Functional complementation permitted only the heterokaryons to survive. A total of 706 putative heterokaryon-derived plantlets were regenerated and 570 survived transplantation to soil. Experimentation was aimed at the introduction of proanthocyanidins (condensed tannins) from sainfoin, a bloat-safe plant, to alfalfa, a bloat-causing forage crop; however, no tannin-positive regenerant plants were detected. Most regenerant plants have shown morphological differences from the fusion parents, although, as expected, all resembled the "recipient" parent, alfalfa. Southern analysis using an improved total-genomic probing technique has shown low levels of sainfoin-specific DNA in 43 out of 158 tested regenerants. Cytogenetic analysis of these asymmetric hybrids has confirmed the existence of euploid (2n=32; 17%) as well as aneuploid (2n=30, 33-78; 83%) plants. Pollen germination tests have indicated that the majority of the hybrids were fertile, while 35% had either reduced fertility or were completely sterile.

A novel method is presented for the synthesis and purification of (+)-2,3-trans-3,4-cis-[4-3H]leucocyanidin. Soluble enzyme extracts from developing barley grains and leaves of the forage legume Onobrychis viciifolia (sainfoin) catalyzed the NAD(P)H-dependent reduction of (+)-2,3-trans-3,4-cis-[4-3H]leucocyanidin to (+)-[4-3H]catechin. NADPH was the preferred substrate. With extracts of barley the rate of reaction with 1 mM NADH was 20% of the rate found with NADPH. With extracts from both tissues there was a broad pH optimum around pH 6.6.

The mechanism of NaCl transport across the epithelium of intact MDCK cysts grown in a collagen gel matrix was investigated. Double-barreled microelectrodes were used to measure basolateral membrane PD (Vbl), transepithelial PD (Vt), and intracellular (Cli) and intralumenal (Clcy) Cl- activities in cysts under different conditions. In a control Ringer's solution (RS), Cli (60 +/- 1 mM) and Clcy (107 +/- 2 mM) exceeded the values corresponding to electrochemical equilibrium across the basolateral membrane and epithelium, respectively. Cli was reduced by superfusing the cysts with a low Cl- RS (Cli, 20 +/- 3 mM), a low Na+ RS (Cli, 40 +/- 4 mM), or by adding amiloride to the control RS (Cli, 46 +/- 1 mM). Cli was unaffected by removal of either K+ or HCO3- from the RS or by adding furosemide or SITS to the control RS. Vbl in the control RS was -50 +/- 2 mV and was affected only by removal from the RS of K+ (Vbl, -31 +/- 3 mV) or HCO3- (Vbl, -29 +/- 4 mV) or by the addition of SITS to the control RS (Vbl, -59 +/- 5 mV). Vt in control RS was -2 +/- 0.2 mV (lumen negative), and was increased by reducing bath Na+ (Vt, -37 +/- 2 mV) but not by reducing bath Cl-. These data indicate that Cl- is secreted in a basolateral to apical direction by the cyst epithelium. Basolateral Cl- transport probably occurs mainly by an electroneutral Cl-/HCO3- exchanger. Transepithelial Na+ transport seems to occur via a paracellular route which appears to be cation selective. These experiments also support the existence, in the basolateral membrane, of a Na+/K+ ATPase, a Na+/H+ exchanger, and possibly a Na+/HCO3-/CO3(2-) transporter.

Recent progress in the semiclassical description of two-electron atoms is reported herein. It is shown that the classical dynamics for the helium atom is of mixed phase space structure, i.e., regular and chaotic motion coexists. Semiclassically, both types of motion require separate treatment. Stability islands are quantized via a torus-quantization-type procedure, whereas a periodic-orbit cycle expansion approach accounts for the states associated with hyperbolic electron pair motion. The results are compared with highly accurate ab initio quantum calculations, most of which are reported here for the first time. The results are discussed with an emphasis on previous interpretations of doubly excited electron states

Starting from the semiclassical dynamical zeta function for chaotic Hamiltonian systems we use a combination of the cycle expansion method and a functional equation to obtain highly excited semiclassical eigenvalues. The power of this method is demonstrated for the anisotropic Kepler problem, a strongly chaotic system with good symbolic dynamics. An application of the transfer matrix approach of Bogomolny is presented leading to a significant reduction of the classical input and to comparable accuracy for the calculated eigenvalues.

30 autopsy cases of gliomas (5 fusiform glioblastomas, 5 multiform glioblastomas, 5 fibrillary astrocytomas, 5 protoplasmic astrocytomas, 5 oligodendrogliomas, 5 spongioblastomas) were studied by means of morphometry. 7 parameters (volume and area parameters of tumor vessels and of tumor cell nuclei, mitoses) were tested for the ascertainament of useful quantitative criteria for the objective differentiation of the gliomas. 210 statistical comparisons were carried out. Significant differences were observed in 32 of them. It could be demonstrated that the parameters of the tumor cell nuclei are most useful in the differentiation of the gliomas. Significant differences do not exist between fusiform glioblastoma--multiform glioblastoma, fusiform glioblastoma--oligodendroglioma, fibrillary and protoplasmic astrocytoma--spongioblastoma.

The evolution of mathematical models of the uptake of the inhaled anesthetic agents has produced increasingly complex models in which researchers have attempted to incorporate more and more data on the effects of anesthetics on the processes of respiration, circulation, and metabolism. One result of this evolution has been to limit the application of these models due to the large amount of data required by the model and the need for a large digital computer to generate a solution. The purpose of this study is to show that a three-compartment linear model, using only the solubility of an anesthetic in water and oil, may br used to predict the uptake of a volatile anesthetic with sufficient accuracy for practical purposes. Only a programmable hand calculator is needed for the solution. Due to the simplicity of this model, compared with previously described models, it should prove useful in understanding the kinetics of gas uptake by the body.

Acute kidney failure was produced in the anesthetized rat by 1 h of complete renal artery occlusion. Kidney function was studied either immediately after release of the occlusion or 1 day later using clearance, micropuncture, histological, and nephron dissection techniques. Polyfructosan clearance was decreased to 5% of normal after temporary occlusion. Proximal tubular pressure (PTP) averaged 13-14 mmHg in normal kidneys, 39 mmHg immediately after release of unilateral occlusion, 19 mmHg 1 day after unilateral occlusion, and 25 mmHg 1 day after bilateral occlusion. The increased PTP reduces the glomerular filtration rate (GFR). Glomerular capillary pressure, estimated from the sum of the stop-flow and arterial plasma colloid osmotic pressures, was not decreased after temporary ischemia. Single-nephron GFR, measured without intratubular pressure control, was only slightly below normal 1 day after bilateral occlusion. Most distal tubules from ischemia-damaged kidneys contained hyaline casts. Tubular obstruction is a major factor in this model of acute kidney failure.

This study examined the effect of blocking proximal tubule lumens on glomerular and early proximal tubular morphology. Single nephrons in the rat kidney were blocked with wax by micropuncture. After one day, one week, or one month of obstruction, the kidneys were fixed with glutaraldehyde by intravascular perfusion, and nephron structure was examined by light and electron microscopy. Following obstruction, glomerular changes developed more slowly than tubular changes. After one day, the only change noted in some glomeruli was the presence of inflammatory cells. The only tubule change upstream to the block was a focal loss of apical microvilli. This is in contrast to the severe damage previously reported (Evan, Tanner: Kidney Int 30: 818-827, 1986) in downstream proximal tubule segments at this time. After one month of obstruction, glomerular size was decreased and the glomerular filtration membrane was abnormal. Tubular cell size was decreased, apical microvilli were lost, basolateral interdigitations were reduced, and mitochondria were fewer and abnormally oriented. Interstitial fibrosis was present. Changes in nephron structure develop slowly after obstruction, perhaps because continued filtration and reabsorption maintain nephron integrity. Eventually, blocked nephrons atrophy, probably because of reduced blood flow, disuse, and inflammatory responses.

Blunt traumatic injury to the biliary tract is rare, and its management is one of the most difficult and challenging problems confronting surgeons. If disruption occurs in the hepatic ducts, occult ductal injury may even go unnoticed. A high index of suspicion is the single most important factor leading to the identification and successful management of these injuries. A patient with massive upper abdominal injuries secondary to blunt trauma is reported on. Intraoperative cholangiography demonstrated bilateral hepatic duct transection. The injury was successfully managed by Roux-en-Y hepatoportal enterostomy, an approach that has not been previously reported. Primary repair or hepatic cholangiojejunostomy is the treatment of choice for hepatic duct injuries. Hepatoportal enterostomy, however, offers a satisfactory alternative in treatment when the patient is unstable or when primary repair is not possible. The literature is reviewed and the pathogenesis, diagnosis and treatment of nonpenetrating injuries to the biliary tract are discussed.

We observed a case of skin avulsion associated with measurement of blood pressure by oscillometry on an anesthetized patient in the prone position. Epidermal shearing may be avoided in the prone patient by either wrapping the antecubital skin underlying the blood pressure cuff with gauze or preventing weight-bearing by the cuffed portion of the arm.

This exploratory study examined the effect of structured teaching about essential hypertension on control of the diastolic blood pressure in individuals with a diagnosis of essential hypertension. Using the pre-test, post-test design, study participants were measured on knowledge about essential hypertension and diastolic blood pressure before and after intervention. Six hypotheses were tested, three related to control of blood pressure, and three related to knowledge about essential hypertension. There was no significant difference between the experimental and control groups on diastolic blood pressure readings at the conclusion of the study. There was a significant difference between the experimental and control groups on post-test scores on knowledge of essential hypertension. It was concluded that structured teaching had no significant effect upon control of the diastolic blood pressure. Instruction increased knowledge, but not blood pressure control.

A new and more sensitive cuprammonium reagent containing strong base has been developed for the colorimetric detection of polyols and sugars. A reagent blank with an absorbance at 280 nm, less than half that of previous methods, was achieved by reducing the cupric ion and ammonia concentrations to 1 mM (from 5 mM) and 0.4 M (from 2.25 M), respectively, and adding 0.2 M sodium hydroxide. Polyols are particularly well suited to the new method, showing up to an eightfold increase in sensitivity. The new reagent is readily applicable to the postcolumn detection of polyols and sugars after high-performance liquid chromatography and is linear over the range 40-180 nmol for D-glucitol (sorbitol) and 10-300 nmol for glucose, using standard HPLC equipment.

We describe a seven-compartment physiologic model of inhalational anesthetic induction with circulatory shunts that was used to simulate inhalational anesthetic induction in children with congenital heart disease. Our conclusions based on this model are that left-to-right (L-R) shunting has little effect on speed of induction; right-to-left (R-L) shunting significantly slows induction of N2O and halothane anesthesia; and adding an L-R shunt to a preexisting R-L shunt will attenuate the slowing of induction caused by the R-L shunt.

This study examined the effects on proximal tubule morphology of blocking single nephrons with paraffin wax for one day, one week, or one month in the rat. Proximal tubule lumens were blocked with a short column of wax using micropuncture. Chronically blocked and control (normal) tubules were fixed by either intravascular or intraluminal perfusion of glutaraldehyde solution. Proximal tubule segments downstream to the wax block were examined by light and transmission electron microscopy. Intraluminal Alcian blue dye, serial sectioning, and nephron microdissection techniques were used to identify nephrons. One day after obstruction, all proximal tubule cells downstream to the block were injured. Some recovery was seen. S1 and S2 segments showed more severe damage than S3 segments. Alcian blue, which normally is excluded from cells, entered the cytoplasm of some damaged S1-S2 cells. After one week of obstruction, the tubule appeared to have reconstituted itself, but cells were less differentiated than normal. One month after obstruction, blocked tubules were atrophied. Tubule cells were simplified and were surrounded by a thickened basement membrane. The results suggest that prolonged proximal tubule blockade produces injury and atrophy of the proximal tubule probably due to ischemia and interruption of normal reabsorptive activity.

The subcellular localization of hexose phosphorylating activity in extracts of pea stems has been studied by differential centrifugation and sucrose density gradient centrifugation. The hexokinase (EC 2.7.1.1) was associated with the mitochondria, whereas fructokinase (EC 2.7.1.4) was in the cytosolic fraction. Some properties of the mitochondrial hexokinase were studied. The enzyme had a high affinity for glucose (K(m) 76 micromolar) and mannose (K(m) 71 micromolar) and a relatively low affinity for fructose (K(m) 15.7 millimolar). The K(m) for MgATP was 180 micromolar. The addition of salts stimulated the activity of the hexokinase. Al(3+) was a strong inhibitor at pH 7 but not at the optimum pH (8.2). The enzyme was not readily solubilized but, in experiments with intact mitochondria, was susceptible to proteolysis. A location on the outer mitochondrial membrane is suggested for the hexokinase of pea stems.

The effects of chronic tubule blockade with paraffin on estimated glomerular capillary pressure (GCP) and single nephron glomerular filtration rate (SNGFR) were examined in anesthetized rats using micropuncture techniques. Blockade of individual proximal and distal tubule lumens for 26 hr resulted in identical reductions in GCP (by about 10 mm Hg). Proximal tubules were also blocked for about 6 hr with and without a hole in the tubular wall just upstream to the block; the hole prevented an increase in proximal tubular pressure. GCP was decreased in the block-plus-hole nephrons, suggesting that an increase in pressure in Bowman's space is not the stimulus leading to a fall in GCP. Blockade of tubules for 26 hr reduced SNGFR from 20.4 +/- 1.0 to 12.0 +/- 0.8 nl . min-1 . 100 g of body wt-1. In conclusion, chronic tubular blockade results in reductions in GCP and SNGFR unrelated to changes in tubular pressure. We suggest that this tubuloglomerular feedback response is related to decreased fluid delivery to the distal tubule macula densa.

Simulations based on physiologic modeling have been used for over twenty years to clarify the pharmacokinetics of inhaled anesthetics. We describe ANSIM, an anesthesia simulator for the IBM PC. ANSIM uses established modeling techniques to create a life-like anesthesia simulator. ANSIM is useful as an educational tool for demonstrating classic concepts of uptake and distribution of inhaled anesthetics.

The present study tested the idea that protein in tubular fluid is responsible for an increase in resistance of Henle's loops in the nephrotic syndrome. Single proximal tubules of anesthetized, normal rats were microinfused with an isotonic equilibrium solution containing 0--250 mg/100 ml bovine serum albumin while proximal tubular pressure (PTP) was continuously recorded. Microinfusion at 16 and 40 nl/min produced significant (p less than 0.01) increments in PTP. There was no significant effect (p greater than 0.50) of albumin. These results suggest that moderate concentrations of albumin in tubular fluid do not cause an increase in resistance to tubular fluid flow.