Tonic activity of submucosal neurons influences basal ion transport

The influence of tonically active submucosal neurons on basal ion transport was studied using sheets of guinea pig ileum set up in flux chambers. Tetrodotoxin evoked an immediate and sustained decrease in short-circuit current that was sustained for 60 minutes compared with control tissues in which basal currents gradually decreased over time. Time-dependent changes in basal short-circuit currents in tissues treated with atropine were not significantly different from control tissues. The decrease in short-circuit current after tetrodotoxin resulted from a greater increase in net chloride absorption than sodium absorption. Changes in net sodium and chloride transport were due to an increase in the mucosal-to-serosal fluxes of these ions. The results suggest that tonic activity of submucosal neurons limits the absorptive capacity of the guinea pig ileum.     

A Modification of Mayer's Tannic Acid-Ferric Chloride Staining Method for Demonstrating Cellular Membranous Systems for Light Microscopy

To observe cellular membranous systems under a light microscope, we modified Mayer's tannic acid-ferric chloride stain method by adding a treatment with hematoxylin after the original procedure. We used the modified tannic acid-ferric chloride (MTA-Fe) stain method to examine kidneys, liver, heart, trachea, epididymides and other organs of rats and dogs. The MTA-Fe stain clearly demonstrated the basement membrane, brush border, basolateral invaginations and cell processes in the kidneys which enabled easy differentiation of the S1 and S3 segments of proximal convoluted tubules. Our technique also demonstrated hepatic cell membranes and bile canaliculi in the liver, cross striations and longitudinal traveling of myofibrils in the heart, cilia of the epithelial cells in the trachea, and stereocilia and terminal bars in the epididymis. The MTA-Fe stain is a convenient method to visualize cellular membranous systems even for light microscopy. The stain has the advantages of using no toxic materials, simple and easy technique, little variation of staining results, and little fading for several months after staining.     

Robert W. Mowry,M.D. Some thoughts about his contributions to the Biological Stain Commission

To observe cellular membranous systems under a light microscope, we modified Mayer's tannic acid-ferric chloride stain method by adding a treatment with hematoxylin after the original procedure. We used the modified tannic acid-ferric chloride (MTA-Fe) stain method to examine kidneys, liver, heart, trachea, epididymides and other organs of rats and dogs. The MTA-Fe stain clearly demonstrated the basement membrane, brush border, basolateral invaginations and cell processes in the kidneys which enabled easy differentiation of the S1 and S3 segments of proximal convoluted tubules. Our technique also demonstrated hepatic cell membranes and bile canaliculi in the liver, cross striations and longitudinal traveling of myofibrils in the heart, cilia of the epithelial cells in the trachea, and stereocilia and terminal bars in the epididymis. The MTA-Fe stain is a convenient method to visualize cellular membranous systems even for light microscopy. The stain has the advantages of using no toxic materials, simple and easy technique, little variation of staining results, and little fading for several months after staining.     

The effects of selenium on the distribution of mercury in the organs of the black bullhead (Ictalurus melas)

Following i.p. mercuric chloride injections, the mercury was deposited primarily in the kidneys. Simultaneous selenium injections prevented mercury induced osmoregulatory failure even though selenium strongly promoted the movement of mercury to the kidneys and its deposition in an approximate 1:1 mercuric selenite ratio. Whole-body retention of mercury was not altered by simultaneous subcutaneous injections of sodium selenite.     

Explanation for constancy of intracellular chloride concentration with respect to membrane potential in guinea pig mammary gland.

1. The relative permeabilities for sodium, potassium and chloride in guinea pig mammary gland slices are determined by means of ion flux studies with radioisotopes. 2. Assuming that there are no significant electrogenic potential components, we calculate permeability ratios PNa/PK = 0.97 and Pc1/PK = 1.25. 3. Substitution of these values in the Goldman equation yields membrane potentials of--15 mV before and--13 mV after ouabain treatment. 4. This small change in membrane potential explains the absence of a significant change in chloride content upon ouabain application, which leads to large changes in intracellular sodium and potassium concentrations.     

Mechanisms of transport of Na+ and Cl- in the lizard colon

1. Ionic fluxes of sodium and chloride across lizard colon mucosa were measured and compared with the electrical characteristics of the tissue under voltage-clamped conditions. 2. In a Ringer-bicarbonate solution there was both a net sodium flux (JNanet) and a net chloride flux (JClnet) from mucosa to serosa. The net flux residual (JR) was near zero, indicating that net sodium and chloride transport is the result of an electrically neutral transport mechanism. 3. In the presence of sodium, the net chloride flux was abolished and the short-circuit current (Isc) and the electrical potential difference (PD) were unchanged. In the absence of chloride the net sodium flux was abolished and the short-circuit current and electrical potential difference were not modified. 4. From an analysis of the effects of the inhibitors, furosemide, amiloride and disulfonic stilbene (DIDS), a plausible model was developed to explain the characteristics of sodium and chloride absorption.     

Proximo-distal specialization of epithelial transport processes within the Xenopus pronephric kidney tubules

The embryonic kidneys of larval aquatic vertebrates such as fish and frogs serve as excellent model systems for exploring the early development of nephric organs. These experimental systems can easily be manipulated by microsurgery, microinjection, genetics, or combinations of these approaches. However, little is known about how physiologically similar these simple kidneys are to the more complex mammalian adult kidneys. In addition, almost nothing is known about proximo-distal patterning of nephrons in any organism. In order begin to explore the physiological specialization of the pronephric tubules along the proximo-distal axis, a combination of uptake assays using fluorescently tagged proteins, LDL particles and dextrans, and an informatics-targeted in situ screen for transport proteins have been performed on embryos of the frog, Xenopus laevis. Genes identified to be expressed within unique subdomains of the pronephric tubules include an ABC transporter, two amino acid cotransporters, two sodium bicarbonate cotransporters, a novel sodium glucose cotransporter, a sodium potassium chloride cotransporter (NKCC2), a sodium chloride organic solute cotransporter (ROSIT), and a zinc transporter. A novel combination of colorimetric and fluorescent whole-mount in situ hybridization (FCIS) was used to precisely map the expression domain of each gene within the pronephros. These data indicate specialized physiological function and define multiple novel segments of the pronephric tubules, which contain at least six distinct transport domains. Uptake studies identified functional transport domains and also demonstrated that early glomeral leakage can allow visualization of protein movement into the pronephric tubules and thus establish a system for investigating experimentally induced proteinuria and glomerulonephritis.     

Excretory function of denervated kidney after inhibition of prostaglandin synthesis and furosemide administration in conscious dogs

The experiments were carried out on unanaesthetized dogs with exteriorized ureters for separate urine collection from the left (denervated) and the right (intact) kidney. The osmolality and concentrations of sodium, potassium, calcium, magnesium, zinc, copper, chloride and creatinine were determined in the plasma as well as in the urine of the two kidneys. The function of the denervated and the innervated kidney was compared prior to and after indomethacin administration (5.0 mg/kg b.w.). The excretory function of both kidneys was also compared after furosemide treatment alone (0.5 mg/kg b.w.) as well as indomethacin pretreatment. Renal denervation increased urine flow rate, calcium and copper excretion. After administration, sodium excretion from the denervated kidney was higher than that from the intact one. Calcium excretion of the two kidneys did not differ significantly, while copper excretion from the denervated kidney was diminished, Furosemide administration after pretreatment with indomethacin did not lead to any difference between the denervated and intact kidney. The results show that renal nerves and prostaglandins participate jointly in the regulation of sodium, copper and calcium excretion. Renal prostaglandins do not change the response of the denervated kidney to furosemide as compared to the intact kidney.     

Acid-base state of blood and accumulation of strontium in tissues of rats

It is shown that the increase of strontium content 1.5, 1.7, 2.0, 1.7, and 5.1 times, respectively, in the liver, kidneys, heart, muscles and bones of poisoned rats leads to the state of subcompensated metabolic acidosis in them. Experimental deepening of metabolic acidosis with per os introducing of chloride acid parallel with intramuscular injection of strontium chloride reduces the accumulation of strontium in the muscles and kidneys 1.6, in the liver - 1.5, in bones - 2.7 times. Introduction of rats into metabolic acidosis state after their poisoning with strontium causes acceleration of mentioned metal elimination process from the muscles and kidneys 1.5 times, from the liver 1.3 times, from the bones 1.4 times, on the 20th day of research. The issues concerning subcompensated metabolic alkaloses observed in rats' blood, poisoned by strontium after per os introducing of sodium hydrocarbon. It is proved that this phenomenon does not influence accumulation of strontium both in the heart and bones, and it does not essentially reduce the accumulation of this element in the muscles, liver and kidneys. Therefore, the experimental results performed evidence that in the case of changing the acid-base state indices of blood in the direction of metabolic acidosis, one could reach the reduction of strontium accumulation in poisoned animals' organism, and acceleration of mentioned metal elimination process. It should be mentioned that metabolic alkaloses does not influence the intensity of the mentioned metal elimination process.     

Reduction of the effects of diuretics or sodium chloride loading in the conscious rat by inhibitors of prostaglandin synthesis

In conscious rats pretreatment with indomethacin or flurbiprofen, two chemically unrelated inhibitors of prostaglandin synthesis, reduced urine volume and sodium excretion induced by four diuretics, acetazolamide, amiloride, bendrofluazide and frusemide, or oral sodium chloride loads. The maximum reduction in sodium excretion was limited to approximately 2 mmol/kg Na⁺ even when sodium excretion was greatly increased. In contrast these inhibitors did not appreciably affect potassium excretion. These results indicate that part of the natriuretic response in the rat to highly and moderately efficacious diuretics and to sodium chloride loading is modified by prostaglandins. We suggest that the lack of effects on potassium excretion indicate that the collecting tubule is the probable site of action.     

Immunomicroscopical localization of human preformed natural antibodies against pig tissues in xenogeneic transplantation

Summary The targets of preformed natural antibodies need to be identified whenever the use of pig organs is considered for human transplantation. In this study we used extracorporeal perfusion of pig organs with human blood, immunocytological techniques and immuno-electron microscopy to identify the targets and the nature of human preformed natural antibodies against pig antigens. The antibodies were found to be mainly of the IgG and IgM type and directed not only against endothelial cells, but also against mesenchymal and epithelial structures. To reproduce an in vivo situation, a Bio-pump was used to xenoperfuse pig kidneys and livers with human fresh oxygenated blood at 37°C, drawn from polycythaemic patients. Biopsies showed a deposition of human IgG and IgM on tubuli and glomeruli of pig kidneys and on endothelial cells of pig livers. Preperfusion of pig liver with human blood for 45 minutes before perfusion of kidneys significantly reduced the deposition of the natural antibodies.     

Paddle cilia and discocilia — Genuine structures?

Kinocilia of epidermal sensory cells in fixed marine Turbellaria often terminate as flattened biconcave discs. The distal part of the ciliary axoneme curves back upon itself forming a 360 degree loop which is enveloped by the plasmalemma. In living animals this structure can be induced by the addition of sodium cacodylate, monobasic sodium phosphate, dibasic sodium phosphate, sucrose, calcium chloride, or formaldehyde to the sea water. Specimens treated with sodium chloride, glutaraldehyde, or osmium tetroxide do not show modified cilia. In animals prepared for EM at low temperature and with a buffered hypotonic fixative less kinocilia are modified than in animals treated with a buffered iso- or hypertonic fixative and at a higher temperature. It is assumed that the unusually shaped cilia, described as "paddle cilia" or "discocilia" in other invertebrates, do not represent a genuine but an artificial structure.     

Direct effects of altered temperature on renal structure and function

Although marked alterations in temperature often accompany ischemic, acute renal failure (ARF), the effects of altered temperature on renal structure and function have received little attention. In the present investigation, isolated rat kidneys perfused at 41 degrees C had extensive tubular damage and decreased function compared to kidneys perfused at 37 degrees C. In contrast, kidneys perfused at 30 degrees C had less tubular damage, and better function, than kidneys perfused at 37 degrees C. Increased temperature caused a 50% reduction in renal ATP (0.46 +/- 0.04 microM/100 mg tissue protein. 37 degrees C vs. 0.26 +/- 0.03 microM/100 mg tissue protein, 41 degrees C; p less than 0.05). The decreased ATP occurred despite reduced sodium reabsorption (129 +/- 8 microM/min/g, 37 degrees C vs. 65 +/- 12 microM/min/g, 41 degrees C, p less than 0.05) and normal renal oxygen consumption (QO2). These results suggest that increased temperature may cause an uncoupling of QO2 and sodium chloride transport, and an increase in nontransport mediated, basal metabolic rate may result in depleted cellular ATP levels and renal tubular cell death.     

Comparison of the effect of temperature on kidney cortex mitochondria from rabbit,dog, pig,and human: Arrhenius plots of ADP-stimulated respiration

The effect of temperature on the rate of ADP-stimulated respiration of mitochondria from dog, rabbit, pig, and human kidney cortex mitochondria was plotted according to the Arrhenius relationship. The temperature at which the plot demonstrated a break was at 15 °C for mitochondria from dog, pig, and human kidneys. The discontinuity occurred at 10 °C or less for mitochondria from rabbit kidneys. This difference suggests that mitochondria from rabbit kidneys undergo a lipid-phase transition at lower temperatures than for other species commonly used in experimental renal preservation. The implications of this difference suggest caution in using results obtained with rabbit kidneys for comparison to results obtained from hypothermic renal preservation of other species kidneys. Apparent fluidization of dog kidney mitochondrial membranes with adamantine abolished the discontinuity in the Arrhenius plot.     

The production of antibacterial tubing, sutures, and bandages by in situ precipitation of metallic salts

Two procedures were used to modify gauze bandages, polyester sutures, silicone tubing, and polyvinyl chloride tubing. In one procedure, the materials were first modified by in situ precipitation of metallic hydroxides and then used to adsorb silver ions. In the second procedure, the materials were soaked in sodium pyrophosphate or sodium chloride, dried, and then soaked in silver nitrate. These procedures produced materials with silver deposited on the surface of the tubing and sutures and both on the surface and within the gauze fibers. The modified materials inhibited the growth of Pseudomonas aeruginosa. Escherichia coli, and Staphylococcus aureus in vitro.     

Influence of augmentation of excretory renal mass on renal function after alpha-receptor blockade

The effect of renal function of an augmentation of the excretory renal mass was investigated in 10 dogs without drug treatment and in 10 animals with alpha-receptor blockade. In the untreated group, augmentation of excretory renal mass by transplantation into the neck of one pair of kidneys isolated from another animal caused the following changes in the kidneys in situ: marked elevation in CPAH, slight decrease in Cinulin, slight diminution of urine excretion and a pronounced fall in sodium excretion. The amount of urine and sodium excreted by the four kidneys was identical with that previously excreted by the two kidneys in situ. In animals with alpha-receptor blockade, augmentation of the excretory renal mass had the following consequences in the in situ kidneys, CPAH, and Cinulin remained unchanged while urine and sodium excretion decreased to the same extent as in the untreated control group. The amount of urine and of sodium excreted by the four kidneys was the same as that excreted by the kidneys in situ, prior to transplantation of isolated kidneys, i.e. before the augmentation of excretory renal mass. It seems that the decrease in sodium excretion of the kidneys in situ was not due to the haemodynamic changes evoked by the load on the circulation; it was rather consequence of some quick, presumably humoral, regulation. The diminution of sodium excretion in the kidneys in situ after augmentation of the excretory renal mass has been ascribed to an increased utilization by the four kidneys of the natriuretic factor(s), i.e. to a diminution in the plasma level of the natriuretic hormone.     

Ammonium chloride potentiation of streptozotocin-induced diabetes in juvenile pigs.

In preparation for direct vital microscopic evaluation of microcirculatory dynamics in the diabetic pig myocardium, we were initially unsuccessful in inducing sustained hyperglycemia in juvenile pigs using streptozotocin according to previously reported methods. Therefore, we modified the technique in a way previously unreported in an effort to improve the success rate of diabetes induction. In the first set of 9 pigs, we followed described methods of intravenous injection with 150 mg/kg streptozotocin. In the second group of 9, the technique was modified. The change was based on human studies with ammonium chloride and animal experiments with alloxan, and consisted of the addition of a period of pretreatment with ammonium chloride. Of the nine pigs not treated with ammonium chloride, only two developed sustained hyperglycemia in excess of 17 mmol/L (300 mg/dl), and only after reinjection with a full dose of streptozotocin within 7 days of the first injection. Conversely, of the ammonium chloride pretreated pigs, eight of nine developed diabetes. We conclude that the addition of ammonium chloride to produce systemic acidosis prior to streptozotocin injection improves the efficacy of the drug.     

Haemolysis of various mammalian erythrocytes in sodium chloride, glucose and phosphate-buffer solutions.

Mouse, rat, rabbit, hamster, cow, pig, sheep, guinea-pig, dog and human erythrocytes were studied. A 0.9% or stronger solution of sodium chloride completely prevented haemolysis; sheep and pig erythrocytes appeared the more fragile, while human and dog erythrocytes were not haemolized in concentrations of 0.4% or more. Haemolysis of human, rabbit, cow, hamster, guineapig, pig and sheep erythrocytes was not observed in solutions of 0.4% or more of glucose. Except for sheep, human and dog erythrocytes, haemolysis was depressed in rate but not completely prevented by phosphate-buffer solution of pH 7.0.     

Oxygen cost of chloride transport in perfused rectal gland ofSqualus acanthias

Summary In the stimulated state, with glucose as substrate, oxygen uptake by the isolated perfused rectal gland is directly related to the rate of chloride secretion. Lactate production is negligible under aerobic conditions in the stimulated gland. A stoichiometric relationship exists between chloride transport and oxygen consumption, with a Cl/O₂ ratio of about 301, resembling that reported for sodium in mammalian kidneys. This ratio remains constant under varying degrees and modes of stimulation. The ratio does not change when the gland is induced to secrete chloride against varying electrochemical gradients by altering the concentration of urea in the perfusate.Established Investigator of the American Heart Association.