bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of a computer simulation program and a traditional laboratory practical class for teaching the principles of intestinal absorption.
Authors: Dewhurst D G , Hardcastle J , Hardcastle P T , Stuart E
Published: December 2017

Here we describe an evaluation of the effectiveness, compared with a traditional laboratory, of an interactive computer-assisted learning (CAL) program, which simulates a series of experiments performed using isolated, everted sacs of rat small intestine. The program is aimed at undergraduate students of physiology and is designed to offer an alternative student-centered learning approach to the traditional laboratory-based practical class. The evaluative study compared two groups of second-year undergraduate students studying a module on epithelial transport: one group worked independently using the CAL program and associated learning materials, and the other group followed a conventional practical class approach, working in the laboratory under supervision. Knowledge gain of each group was measured by means of a test consisting of a range of question types (e.g., short-answer factual, calculation, interpretation) given to students before and after the module. Student attitude to both approaches was assessed by questionnaire, and the resource requirements were also compared. It was found that the knowledge gain of both groups of students was the same, that students had a positive attitude toward using CAL programs of this type, and that the cost of the conventional laboratory-based approach was five times greater. The potential for integrating CAL programs into the undergraduate curriculum is discussed.
Incorporating active learning into a traditional curriculum.
Authors: Huang A H , Carroll R G
Published: December 2017

During the past three academic years, "self-learning exercises" (SLEs) have been incorporated into the Medical Physiology course for first-year students at the Morehouse School of Medicine. Roughly 20-30% of the material covered in the course is presented to the students in the form of these exercises, instead of in lectures. The exercises are intended to help the students develop skills in active learning and problem solving. Formal analysis of student performances on multiple-choice exam questions showed that the SLEs did not significantly impair learning evaluated by this traditional means. Student feedback was strongly negative the first year and prompted a number of revisions in the format of the SLEs, which seem to have made them more palatable, without negating their emphasis on active learning and application of material.
Laboratory exercise using "virtual rats" to teach endocrine physiology.
Authors: Odenweller C M , Hsu C T , Sipe E , Layshock J P , Varyani S , Rosian R L , DiCarlo S E
Published: December 2017

Animal experimentation is limited in many curricula due to the expense, lack of adequate animal facilities and equipment, and limited experience of the teachers. There are also ethical concerns dealing with the comfort and safety of the animals. To overcome these obstacles, we developed a "dry laboratory" using "virtual rats." The "virtual rat" eliminates the obstacles inherent in animal experimentation, such as inadequate budgets, as well as avoiding important animal rights issues. Furthermore, no special materials are required for the completion of this exercise. Our goal in developing this dry laboratory was to create an experience that would provide students with an appreciation for the value of laboratory data collection and analysis. Students are exposed to the challenge of animal experimentation, experimental design, data collection, and analysis and interpretation without the issues surrounding the use of live animals.
Respiratory physiology teaching: determination of residual volume by applying the indicator-dilution technique.
Authors: Heller H , Granitza K , Eixmann B
Published: December 2017

Apart from the current teaching of spirometric methods in laboratory courses on respiratory physiology, we have included an experiment in which medical students determine their own residual volume by applying the indicator-dilution technique. For hygienic reasons we used a bag-in-the-box system to dilute helium within alveolar space by performing the single-breath method. Although each participant independently underwent only one single-breath maneuver, we gained a reliable relationship between residual volume and subjects' height and body weight in 68 female (r = 0.6, P < 0.0001) and 99 male (r = 0.42, P < 0.0001) students. From this successful outcome and with the opportunity to discuss the limitations of the single-breath method as well, we inferred that this experiment affords a transparent and instructive approach to interpreting the determination of lung volumes on the basis of the indicator-dilution technique.
A theoretical foundation for discovery learning.
Authors: Svinicki M D
Published: December 2017

The processes of discovery learning are well grounded in current theories of learning. This article describes how cognitive learning theory supports the components of the discovery learning model. Three main characteristics of discovery learning that relate it to cognitive theory are 1) an emphasis on active learning, 2) the development of meaningful learning, and 3) the capacity to change attitudes and values toward the subject and the self as problem solver.
Use of brook char (Salvelinus fontinalis) physiological responses to stress as a teaching exercise.
Authors: Diouf B , Rioux P , Blier P U , Rajotte D
Published: December 2017

Fish hematological changes during osmotic and cold stress are used to introduce the physiological reactions of the animal to an acute stress. Brook char (Salvelinus fontinalis) were subjected to 1 h of stress before being anesthetized and having blood taken from their caudal vein. Glucose, hemoglobin, hematocrit, and osmolarity were determined in the blood samples. Analyses showed that glucose concentration tends to increase and hematocrit tends to decrease in stressed fish. Changes in hemoglobin concentration occurred only in cold-stressed fish. A rise in blood glucose concentration is the result of cortisol secreted by the hypothalamic-pituitary-adrenal axis. The glucose produced is used as an osmolyte or energy source to resist or combat the stress. In stressed fish, changes in hematocrit could be the result of the osmoconcentration of the blood plasma, as shown by the increase in osmolarity for the same group. In cold-stressed fish, a decrease in hemoglobin concentration could be the result of hemodilution by body cell water.
ARE CURRENT TEXTBOOKS GOOD ENOUGH FOR PHYSIOLOGY EDUCATION? FOR EXAMPLE, THE ECL CELLS ARE MISSING
Authors: Waldum Camilla , Zhao Chun-Mei , Chen Duan
Published: December 2017

Current textbooks are believed to provide an updated knowledge. Medical students usually read the textbooks but not the literature that contain the original research articles and reviews. Here, we examined the gap between the current textbooks and literature with the enterochromaffin-like (ECL) cells as an example. A total of 70 textbooks that were published for medical education during the last 10 yr was examined. The literature has been searched mainly from the Internet. We found that most textbooks (59 of 70) fail to mention the ECL cells. Due to the lack of information on the ECL cells, the mechanisms behind gastric acid secretion are described variously from book to book. However, up to the year 2000, 574 research articles and reviews have been published on the various aspects of the ECL cells. The role of the ECL cells in the regulation of the acid secretion has been well demonstrated for more than 20 years. The fact that the textbooks are out of date cannot be explained by the time required to write and publish them. Therefore, we question whether or not the current textbooks are good enough for physiology education and suggest both teachers and students read not only the textbooks, but also utilize the other sources such as the Internet to find and fill the gaps between the textbooks and literature. This is one of the approaches of problem-based learning.
Prolonged potentials in gastrointestinal muscles induced by calcium chelation
Authors: Prosser C. L. , Kreulen D. L. , Weigel R. J. , Yau W.
Published: December 2017

When stomach muscles of skate, toad, or frog or intestinal muscle of cat are treated with Ca-free physiological solutions containing 2-5 mM EGTA or EDTA, spontaneous spikes and slow waves disappear reversibly. With continued treatment, depolarization of 25-30 mV from resting potentials of -65 mV occurs and rhythmic prolonged potentials of several seconds duration appear. They show rapid depolarization to near zero and rapid repolarization and they may continue for several hours. The prolonged potentials disappear when Na is replaced by Li, Tris, or choline. They are insensitive to TTX. The EGTA-induced waves are abolished by Mn, Co, La, verapamil, and D 600. After 10-15 min in 5 mM EGTA, voltage-current and abolition of anomalous rectification. It is concluded that when bound Ca is removed by a chelator, nonspecific reduction in resistance occurs and Na ions may enter rhythmically through channels normally used by Ca.
Aerobic glycolysis in bone: lactic acid production by rat calvaria cells in culture
Authors: Felix R. , Neuman W. F. , Fleisch H.
Published: December 2017

Considerable data have been accumulated on aerobic glycolysis of intact bone preparations. To test whether aerobic glycolysis is a feature of the bone cells themselves or of the localized conditions within an intact bone, cells isolated from rat calvaria were cultured and the effect of several factors on lactate production was determined. These cells drastically decreased lactate production when the pH in the culture medium was lowered, changing from 100 to 20 percent for a pH shift from 7.4 to 6.75. L-lactate inhibited its own formation by 40 percent at 20 mM. Parathyroid hormone (PTH) (820 U/mg) at a concentration ranging from 0.2 to 5.0 U/ml stimulated slightly the lactate production in a log-linear response, the ratio treated over control changing from 1.1 to 1.3. The maximal stimulation is observed at pH 7.0. Isolated cells respond qualitatively the same as did intact calvaria. Quantitatively, there were significant differences: notably, a smaller response to parathyroid hormone and a higher sensitivity to lowered pH.
Energy relationships between cytosolic metabolism and mitochondrial respiration in rat heart
Authors: Nishiki K. , Erecinska M. , Wilson D. F.
Published: December 2017

Isolated rat heart was perfused with Langendorff's retrograde perfusion method, while the oxygen consumption and the left ventricular pressure were monitored continually. The steady-state contents of metabolites in the cardiac tissue, freeze clamped under various work-load conditions, were determined and the concentrations of free cytosolic ADP and AMP were calculated from the near equilibrium in creatine phosphokinase and adenylate kinase reactions. Increasing respiratory rate with increasing load was accompanied by a fall in the cytosolic free [ATP]/[ADP][Pi] but little change in the mitochondrial free [NAD+]/[NADH]. The free energy of ATP hydrolysis was calculated from the concentrations of the adenine nucleotides and compared with the values computed from the measured turnover number for cytochrome c and redox state of the mitochondrial NAD couple according to a mathematical model. The agreement between the two values was good over a wide range of metabolic conditions, which provides further support for the proposed near-equilibrium model of mitochondrial respiration with control exerted at the cytochrome oxidase-oxygen reaction.
Ephaptic transmission in squid giant axons
Authors: Ramon F. , Moore J. W.
Published: December 2017

Some characteristics of ephaptic transmission of action potentials were investigated with squid giant axons. For these studies two isolated axons were placed side by side or, on occasion, a single long axon was looped to form an "ephapse" between the axon trunk and one of its main branches. Extracellular potentials measured adjacent to axons surrounded by a very restricted volume of liquid ranged up to 80 mV in magnitude and had a shape similar to that of the membrane current. Intracellular records of the same axon regions show small voltage deflections; however, the transmembrane voltage (Vm = Vi - Vo) has the appearance of normally propagated action potentials. Ephaptic transmission of action potentials is possible when the ephaptic region is submerged in oil, as well as when the region is immersed in low-calcium solutions. When the speed of the propagated action potential is lowered by replacing the normal artifical seawater (ASW) with low-sodium ASW, some ephaptic effects are enhanced. It is concluded that in regions in which axons are confined by restricted extracellular volume, the large extracellular voltage changes arising during the passage of an action potential in one can cause ephaptic excitation in another.
Sarcomere motion in isolated cardiac cells
Authors: Rieser G. , Sabbadini R. , Paolini P. , Fry M. , Inesi G.
Published: December 2017

Computerized image-analysis techniques have been employed to examine the sarcomere dynamics of isolated mammalian cardiac myocytes. The cells were prepared by perfusion of adult rabbit hearts with hyaluronidase-collagenase solutions; they exhibited phasic contractions in the presence of 10(-6) M Ca2+. The dissociated cells were visualized by phase microscopy and a video camera interfaced in a minicomputer. Digitized cell images were processed by an algorithm utilizing signal averaging and contrast enhancement to yield data showing individual sarcomere position and shortening vs. time, so that patterns of sarcomere activation could be observed in spontaneously contracting cells. Compared to records of whole-cell shortening and of striation displacement, computerized image analysis provided a much more faithful indication of time course and sequence of sarcomere shortening. Spontaneously contracting cells showed sequential sarcomere shortening beginning at one end and propagating longitudinally with a constant velocity, typically at 100--150 micron/s for beat rates of 40 min-1. Velocities of initial sarcomere shortening appeared to increase with elevated Ca2+. These observations are consistent with a regenerative mechanism of calcium-induced calcium release.
Determination of unidirectional fluxes of phosphate across plasma membrane in isolated perfused rat liver
Authors: Sestoft L. , Kristensen L. O.
Published: December 2017

In livers labeled with 32P by intraperitoneal injection 18 h before perfusion the unidirectional efflux of inorganic phosphate (Pi) was a linear function of the concentration of Pi in the intracellular exchangeable pool of Pi (range, 0--1 mumol/g liver). Calculations of the efflux were based on determination of the specific radioactivity of Pi in the intracellular exchangeable pool of Pi by a new method. Zero efflux was found at biopsy concentrations of Pi of about 1.8 mumol/g liver. The influx of Pi was calculated from the efflux and the net uptake of Pi; it was found to be related to the extracellular concentration of Pi in a fashion that was approximately linear. An increase in the concentration of Pi in the input medium gave rise to an increased influx and, with a time delay, also to a gradually increasing efflux. The increased efflux could be explained as a consequence of an increased intracellular concentration of exchangeable Pi.
pH regulation in barnacle muscle fibers: dependence on intracellular and extracellular pH
Authors: Boron W. F. , McCormick W. C. , Roos A.
Published: December 2017

Intracellular pH (pHi) regulation was studied in acid-loaded barnacle muscle fibers by monitoring recovery of pHi with a pH-sensitive microelectrode. By multiplying the rate of pHi recovery by total intracellular buffering power, the acid extrusion rate was obtained. The acid extrusion rate was greatest at low values of pHi, and declined toward zero as pHi approached normal levels. It increased as the extracellular pH (pHo) was raised either by increasing external [HCO3] ([HCO3]o) at constant PCO2 or by decreasing PCO2 at constant [HCO3]o, but more so in the former case than in the latter. These observations suggest that pHo per se is an important determinant of the acid extrusion rate, but that raising [HCO3]o by itself also stimulates acid extrusion. This would be expected if acid extrusion involves the inward movement of HCO3. When fibers were exposed to HCO3-containing solutions at very low or very high pHo, pHi drifted downward or upward, respectively; thbe drifts were inhibited by 4-acetamido-4' isothiocyanostilbene-2,2' disulfonic acid (SITS). Our results are discussed in terms of possible mechanisms of acid extrusion.
Stretch-induced growth in chicken wing muscles: a new model of stretch hypertrophy
Authors: Holly R. G. , Barnett J. G. , Ashmore C. R. , Taylor R. G. , Mole P. A.
Published: December 2017

A new model of stretch-induced growth is evaluated in four chicken wing muscles stretched to different extents by a spring-loaded tubular assembly. Muscles grew in length and cross section in proportion to the extent to which they were stretched. Longitudinal growth was essentially completed within 1 wk, while muscles grew in cross section through at least 5 wk of stretch. The muscles were neither denervated nor immobilized, and muscle activity as measured by EMG was not increased. Oxidative enzyme activities increased substantially with stretch in the patagialis (PAT), a twitch muscle, but were relatively unchanged in the slow-tonic anterior latissimus dorsi (ALD). Stretch altered mitochondrial enzyme proportions in the PAT, but had little effect in the ALD. Capillary density was unchanged with stretch in the PAT, but decreased in the ALD. Capillary density was unchanged with stretch in the PAT, but decreased in the ALD. Capillary-to-fiber ratio, however, increased in both muscles. We conclude that muscles grow and adapt enzymatically due to stretch, but that these responses are dissimilar in twitch and tonic muscles.
cAMP-stimulated cation cotransport in avian erythrocytes: inhibition by "loop" diuretics
Authors: Palfrey H. C. , Feit P. W. , Greengard P.
Published: December 2017

The effect of a series of diuretically active substituted 3-aminobenzoic acid derivatives and related compounds was investigated on a cyclic AMP-activated Na+-K+ cotransport system in avian erythrocytes. A good correlation between the diuretic potency of this class of "loop" diuretics in the dog and their inhibition of cation cotransport in turkey erythrocytes was found. Selected thiazide-type diuretics were found to be ineffective. The most active compound tested (3-benzylamino-4-phenylthio-5-sulfamoylbenzoic acid) had an effective dose50 of 4.6 x 10(-8) M in the avian system, and was about 5 times more potent than bumetanide and 500 times more potent than furosemide in this regard. The diuretics appear to interact directly with the cation transport system itself, and not with some antecedent step in the sequence of events from intracellular cAMP accumulation to stimulation of transport. The compounds tested did not appear to compete at Na+- or K+-binding sites on the transport system. The similarity in the structure-activity relationship of these agents in the avian erythrocyte and the kidney suggests that the avian erythrocyte may be a useful model for analysis both of the diuretic-sensitive transport system of the mammalian kidney, and of the molecular mechanism of loop diuretic action.
Effects of organic solutes on transmural PD and Na transport in freshwater prawn intestine
Authors: Wyban J. A. , Ahearn G. A. , Maginniss L. A.
Published: December 2017

Addition of 1 mM L-alanine or 0.5 mM D-glucose to the mucosal surface of freshwater prawn (Macrobrachium rosenbergii) intestine rapidly increased transmural potential difference (PD) (serosa became more positive). Luminal perfusion with 1 mM D-alanine did not affect the spontaneous PD. Addition of 10 mM serosal L-alanine induced a slow elevation in PD with the same polarity as shown with luminal alanine. The alanine-evoked PD was a hyperbolic function of luminal alanine concentration and was reduced to zero with 0.5 mM serosal ouabain. Luminal 1 mM L-alanine stimulated JNanet from mucosa to serosa from 1.29 +/- 0.45 to 2.83 +/- 0.66 mu mol . cm-2 . h-1. Extensive metabolic breakdown of L-alanine and D-glucose occurred during intestinal transit resulting in a small, but significant, net flux of unaltered sugar from lumen to blood (103.9 +/- 23.1 nmol . cm-2 . h-1) and an absence of net amino acid flow. While 1 mM alanine influx into the epithelium was largely Na dependent, quantitatively insigificant amounts of Na were cotransported into the cell with amino acid (approx 52 nmol . cm-2 . h-1) to account for increased JNanet observed in the presence of this organic solute. Electrogenic effects of luminal nutrients result from their catabolism to fuels for the basolateral Na pump, Increased Na-pump activity stimulates Na entry into the cell by allosteric apical carrier proteins and enhances transmural PD by accelerating net movement of cation across the tissue.
AMP deamination and IMP reamination in working skeletal muscle
Authors: Meyer R. A. , Terjung R. L.
Published: December 2017

The extent of purine nucleotide cycle (PNC) turnover [i.e., the cyclic deamination of adenosine 5'-monophosphate (AMP) and reamination of inosine 5'-monophosphate (IMP)] was estimated in fast-twitch muscle of pentobarbital sodium anesthetized rats during in situ stimulation, by the accumulation of excess IMP after blocking IMP reamination with the adenylosuccinate synthetase inhibitor, hadacidin. Sodium hadacidin (100 mg/kg, iv) did not alter IMP production and was effective in blocking IMP reamination by 80%. Hadacidin had no effect on IMP levels in muscle at rest or during mild stimulation (1 Hz for 30 min). During 30 min of more intense stimulation (5 Hz), hadacidin treatment resulted in a threefold increase in IMP in the fast-twitch white gastrocnemius section, but had no effect in the fast-twitch red section. The IMP in the fast-twitch white section did not accumulate over the 30-min period, but was produced initially (within 5 min) and kept for being reaminated by hadacidin. Thus, the IMP formation and IMP reamination evident in the saline-injected animals did not occur concurrently. This suggests that the IMP reamination arm of the PNC is not an important pathway in "working" muscle.
Transduction mechanisms in carotid body: glomus cells, putative neurotransmitters, and nerve endings
Authors: Eyzaguirre C. , Fidone S. J.
Published: December 2017

Carotid body chemoreceptors are activated by low PO2, high PCO2, acidity, increased temperature, and tonicity. These receptors are important in homeostasis and mediate their reflex effects on the CNS through sensory discharges of the carotid (sinus) nerve. The receptor complex is formed by glomus (type I) cells and carotid nerve endings, which, morphologically, appear to form a sensory synapse. The junction between glomus cells and nerve endings is enveloped by processes of sustentacular (type II) cells. The mechanisms of chemoreceptor transduction are complex; there is no agreement about the identity of the primary receptor element (glomus cell or nerve terminal) or what mechanisms are responsible for the onset of the sensory discharge in the carotid nerve. There is increasing evidence that integrity of the glomus cell is essential for normal transduction and that the receptor synapse described by morphologists may be functionally active. There is no conclusive evidence, however, that the glomus cell is the primary site of sensory transduction. Stimuli act on the glomus cell to release "transmitter" and/or "modulator" substances; but it is unknown if the released chemicals are directly responsible for the accompanying change in sensory impulse frequency or merely modify an already ongoing discharge. Interactions between glomus cells and nerves may be complicated enough to make it very difficult to resolve this question.
Effects of stimulation frequency on potassium activity and cell volume in cardiac tissue
Authors: Browning D. J. , Strauss H. C.
Published: December 2017

We examined the effects of increases in stimulation frequency on the intracellular potassium activity and cell volume of feline and canine Purkinje fibers and feline ventricular fibers. Intracellular potassium activity was measured with potassium liquid ion exchanger microelectrodes in single- and double-barrel configuration. Cell volume was measured with an electrophysiologic technique. From single-barrel electrode recordings, intracellular potassium activities (means +/- SE) in feline Purkinje fiber, feline papillary muscle, and canine Purkinje fiber are 117.1 +/- 5.4, 87.3 +/- 3.1, and 112.4 +/- 4.4 mM, respectively, with [K]o (external potassium concentration) of 5.4 mM. Thus direct evidence for a 30-mM gradient in intracellular potassium activity from Purkinje fiber to ventricular muscle is presented. Intracellular potassium activity did not change in any tissue studied during or after 3 min of 4 Hz stimulation. The lack of change is partially explained by a reversible 2% decrease in cell volume induced by the stimulation protocol. Net losses greater than 2% seen in earlier studies probably reflect either differences between perfused vs. superfused preparations or functional sequestration of cellular potassium involved in transfers during frequency changes.
Myosin light chain phosphorylation associated with contraction in arterial smooth muscle
Authors: Driska S. P. , Aksoy M. O. , Murphy R. A.
Published: December 2017

The hypothesis that Ca2+ initiates contraction in smooth muscle by activating an endogenous myosin light chain kinase (MLCK) that phosphorylates the 20,000 dalton light chain (LC 20) of myosin was tested in tissues prepared from the media of swine carotid arteries. Unstimulated tissues with low levels of tone exhibited low levels of phosphorylated LC 20. On stimulation with a high-K+ physiological salt solution containing 1.6 mM CaCl2, LC 20 phosphorylation increased to 0.6 mol P/mol LC 20 within 30 s. This increase preceded force development, which required 2-4 min to attain a maximum steady-state value of 3.34 +/- 0.15 (SE) X 10(5) N/m2. These results support the hypothesis, as the stimulus was submaximal for the preparation. However, LC 20 phosphorylation declined significantly from its peak value before steady-state force was attained, reaching near control levels after 10 min of stimulation. The results suggest that Ca2+-stimulated LC 20 phosphorylation is an important physiological control mechanism but that additional factors are involved in the maintenance of tonic isometric force.
Role of calcium and cAMP in the regulation of rat submandibular mucin secretion
Authors: Quissell D. O. , Barzen K. A. , Lafferty J. L.
Published: December 2017

The role of cellular adenosine 3',5'-cyclic monophosphate (cAMP) and calcium in the secretion of [14C]glucosamine-labeled mucins by rat submandibular acinar cells was studied. cAMP appeared to be involved, since 1-methyl-3-isobutylxanthine potentiated the secretory response and cellular cAMP levels increased dramatically following adrenergic stimulation. Furthermore, cAMP analogues were able to elicit a secretory response in the absence of beta-adrenergic receptor activation. Cellular calcium was required for mucin secretion at the level of cAMP action; depletion of cellular calcium by pretreatment with EGTA inhibited the secretory response to both adrenergic stimulation and exogenous cAMP addition, but pretreatment with EGTA did not alter the rise in cellular cAMP induced by norepinephrine. Extracellular calcium was not required to elicit mucin secretion, nor could secretion be elicited by means of the calcium ionophore A23187 alone. However, extracellular calcium may have an important biological role in mucin secretion, since cholinergic receptor activation and alpha-adrenergic receptor activation in conjunction with beta-adrenergic receptor activation potentiated mucin release. In addition, the calcium ionophore A23187 potentiated mucin release following cAMP analogue addition.
Role of calcium in cholinergic and adrenergic mechanisms of eccrine sweat secretion
Authors: Sato K. , Sato F.
Published: December 2017

The role of Ca2+ in eccrine sweat secretion was studied using isolated cannulated monkey palm eccrine sweat glands in vitro. Removal of Ca2+ from the incubation medium promptly abolished sweat secretion induced by methacholine or phenylephrine. In contrast, isoproterenol-induced sweat secretion lasted from 40 to 220 min in a Ca2+-free medium. The methacholine-induced maximal sweat rate was a hyperbolic function of the Ca2+ concentration in the bath and reached a plateau at 1 mM Ca2+. Higher Ca2+ concentrations rather suppressed the secretory rate. The Ca2+ ionophore A23187, but not X537A, at 3 X 10(-6) M induced copious prolonged sweat secretion after a latent period of 10 min. A23187-induced sweat secretion was not inhibited by either atropine or propranolol. D 600 (methoxyverapamil) at 10(-3) M inhibited sweat secretion induced by methacholine or by isoproterenol, although the latter lasted longer than methacholine sweating (20 vs. 5 min) in the presence of D 600. The data support the notion that Ca2+ influx into the cell plays a crucial role in cholinergic and alpha-adrenergic sweating, whereas a partial supply of Ca2+ for isoproterenol-induced sweating is derived from an intracellular store.
By the numbers...
Authors: Fishman Alfred P. , Berne Robert M. , Morgan Howard E.
Published: December 2017

Editorial on statistical methods as discussed by the APS Publications Committee.
Myosin light chain phosphorylation-dephosphorylation in mammalian skeletal muscle
Authors: Manning D. R. , Stull J. T.
Published: December 2017

Phosphorylation of the myosin light chain 2 (LC2) subunit was examined in rat fast-twitch and slow-twitch skeletal muscles in response to repetitive stimulation at 23 and 35 degrees C and on incubation of fast-twitch skeletal muscle with isoproterenol. After a 1-s tetany at 35 degrees C, LC2 phosphate content in extensor digitorum longus muscle increased rapidly and transiently from 0.21 to 0.51 mol phosphate/mol LC2. This pattern of phosphorylation was similar to that observed at 23 degrees C. Increases in LC2 phosphate content were dependent on the frequency and duration of stimulation. In soleus muscle LC2 phosphate content was minimal following a 1-s tetany but increased markedly following more prolonged tetanies. On incubation of extensor digitorum longus muscle with isoproterenol (20 microM), LC2 phosphate content did not change, whereas phosphorylase a levels increased. A positive correlation existed between LC2 phosphate content and potentiation of peak twitch tension in both types of muscles, suggesting a physiological function for LC2 phosphorylation.
Influence of glucose on Ehrlich cell volume, ion transport, and membrane potential
Authors: Laris P. C. , Henius G. V.
Published: December 2017

Incubating Ehrlich ascites tumor cells with 10 mM glucose at room temperature resulted in the following changes. The cells shrank, reaching a minimum volume after 1 h. The decrease in cell volume was 50–90% inhibited by 1 mM furosemide. The mmol K+ and Cl-/mg dry wt decreased, and mmol Na+/mg dry wt increased over the 1 h incubation. The net loss of KCl was inhibited by 1 mM furosemide. Immediately after the addition of glucose, the influx of 86Rb sensitive to ouabain decreased, whereas the influx sensitive to furosemide increased. The total influx of 86Rb with glucose was similar to that of controls. The effluxes of 86Rb and 36Cl increased immediately after the addition of glucose. These effluxes did not increase, however, in the presence of 1 mM furosemide. Initially the ouabain-sensitive Na+ efflux was not changed with glucose, but the ouabain-insensitive Na+ efflux decreased. Furosemide (1 mM) did not influence Na+ efflux. With time the ouabain-sensitive Na+ efflux increased as cellular Na+ levels rose so that at 1 h the ouabain-sensitive Na+ efflux from glucose-treated cells was 2.5–3 times that of control cells. The potential difference across the membrane gradually became more negative by apprdoximately 25 mV, reaching a maximum after 1 h. The hyperpolarization was reversed by 1 mM ouabain. The changes in ionic fluxes on the addition of glucose are compared with changes in ionic fluxes seen during volume regulation.
Microfilaments and the hydrosmotic action of vasopressin in toad urinary bladder
Authors: Hardy M. A. , DiBona D. R.
Published: December 2017

The role of cytoskeletal elements in the antidiuretic hormone (ADH)-induced hydrosmotic response of the toad bladder has been investigated with the microfilament and microtubule disrupting agents, cytochalasin B and colchicine. In the presence of a transmural osmotic gradient, addition of ADH resulted in a measured osmotic permeability coefficient (Pf) of 190 +/- 12 micrometers/s; preincubation with cytochalasin B (2 x 10(-5) M) reduced this value to 138 +/- 6. However, ADH-induced diffusional permeability to tritiated water (PD) was enhanced by 17% with cytochalasin B. The toxin caused a pronounced vacuolation in granular cells only when both ADH and an osmotic gradient were present. This led to experiments with brief glutaraldehyde fixation after hormonal stimulation but before osmotic gradient imposition. Subsequent osmotic flow measurements yielded Pf values of 180 +/- 18 and 210 +/- 7 micrometers/s for ADH and ADH + cytochalasin B, respectively. With gradient imposition only after tissue fixation, no structural alterations were found. We conclude that cytochalasin B inhibition is through disruption of the intracellular portion of the pathway for osmotic flows. Comparable experimentation with colchicine indicated that its interference is primarily with ADH action on the mucosal membrane itself. These results may bear on the significance of particle aggregation in the mucosal membrane, which has been thought to indicate ADH-induced water channels but which is reported to be precluded by cytochalasin B treatment.
The spectrin membrane skeleton of normal and abnormal human erythrocytes: a review
Authors: Goodman S. R. , Shiffer K.
Published: December 2017

The erythrocyte membrane skeleton composed of spectrin, actin, and several other proteins is essential for the maintenance of the erythrocyte shape, reversible deformability, and membrane structural integrity in addition to controlling the lateral mobility of integral membrane proteins. In this review, we shall give an historical development of the current model of the erythrocyte membrane skeleton. We will then describe how the experimental technology developed to study the normal membrane skeleton has paved the way for the recent identification of alterations of skeletal protein interactions in hereditary spherocytosis, hereditary elliptocytosis, and hereditary pyropoikilocytosis. We will conclude with a discussion of some of the more exciting and promising directions for future research that are currently being initiated in this vanguard field of cell biology.
Trophic response of rat brown fat by glucose feeding: involvement of sympathetic nervous system
Authors: Sundin U. , Nechad M.
Published: December 2017

Brown adipose tissue has earlier been suggested as an important site of the diet-induced thermogenesis that results from cafeteria feeding in rats. The aim of the present communication has been to see if any defined component of this diet can mimic the effects of the diet on the trophic response of brown fat and if these effects are mediated by the sympathetic nervous system. Rats fed a lipid emulsion did not show hypertrophy of brown adipose tissue. Rats fed a glucose solution, whether voluntarily or by force feeding, showed a clear trophic response of brown fat, as seen by the morphology of the tissue and its increased wet weight, increased protein content, increased total and specific cytochrome c oxidase activity, and increased mitochondrial guanosine diphosphate binding. Chemical sympathectomy of young rats by guanethidine prior to glucose feeding impaired the glucose-induced effects on brown fat. beta-Adrenergic blockade in adult rats also tended to depress the glucose effect. Consequently we conclude that chronic glucose ingestion can mimic cafeteria feeding with respect to the trophic response of brown fat and that an intact sympathetic nervous system is required for the mediation of the glucose effect to the brown adipose tissue.
Microfluorometric monitoring of pHi in cultured heart cells: Na+-H+ exchange
Authors: Piwnica-Worms D. , Lieberman M.
Published: December 2017

Continuous measurement of intracellular pH (pHi) should enhance the likelihood of defining the mechanisms of pHi regulation in actively contracting preparations of cardiac muscle. A filter microfluorometric technique was adapted for use with growth-oriented embryonic chick heart cells to continuously monitor changes in the fluorescence intensity of the pH-sensitive chromophore 6-carboxyfluorescein, generated in situ. Data pertaining to the direction and the rate of pHi changes assisted in thermodynamically characterizing and ascertaining net kinetic parameters of a Na+-H+ exchange mechanism. Imposing an outward Na+ gradient across the cardiac cell membrane rapidly (t 1/2 = 44 s) induced cytosolic acidification, whereas an inward Na+ gradient produced cytosolic alkalinization (t 1/2 = 40 s). Amiloride (10(-3) M) caused the cytoplasm to acidify (t 1/2 = 46 s) and also reversibly blocked the acidification induced by low extracellular Na+. These results are consistent with the presence of a rapid Na+-H+ exchange mechanism in the cardiac cell membrane. Further investigations are required to characterize the involvement of Na+-H+ exchange in pHi regulation and to differentiate the effects of Na+-H+ exchange from other ion gradient-coupled mechanisms, e.g., Na+-Ca2+ exchange.
Inhibition of prostaglandin biosynthesis in renal (MDCK) cells by cAMP
Authors: Hassid A.
Published: December 2017

Page C369: A. Hassid. “Inhibition of prostaglandin biosynthesis in renal (MDCK) cells by cAMP.” Page C369: in the abstract, the first sentence should read: Cultured renal tubular cells (MDCK) have many of the biological properties of cortical medullary tubular epithelial cells, including the ability to synthesize prostaglandin E2 (PGE2) as the major arachidonate metabolite. Page C373: Table 3 should read as follows:

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Ammonia selectively stimulates neutral amino acid transport across blood-brain barrier
Authors: Mans A. M. , Biebuyck J. F. , Hawkins R. A.
Published: December 2017

The response to increased blood NH4+ of three blood-brain barrier transport systems, which are altered after portacaval anastomosis, was studied. NH4+ acetate was infused for 4 or 22 h to raise blood and brain NH4+, and brain glutamine, to levels similar to those observed after portacaval anastomosis. While brain glutamine content was much higher (16–20 mumol/g) than normal (6 mumol/g) at both times, the permeability of the blood-brain barrier to the neutral amino acid [14C]tryptophan was greater only after 22 h of infusion. After discontinuing the infusion for 5 h, tryptophan transport returned to normal, whereas brain glutamine remained elevated (13 mumol/g). Thus there seemed to be no relationship between the rate of transport and glutamine content. The permeability to [14C]sucrose was unaltered, showing that the integrity of the blood-brain barrier was maintained. Other changes that are characteristic of portacaval shunting, such as decreased basic amino acid ([14C]lysine) and monocarboxylic acid (3-[14C]hydroxybutyrate) transport, were not reproduced by 22 h of infusion. The results demonstrated that the continued presence of NH4+ could be responsible for the change in at least one of the transport systems that are affected by portacaval shunting.
Effects of diabetes on cardiac lysosomes and protein degradation
Authors: Chua B. H. , Long W. M. , Lautensack N. , Lins J. A. , Morgan H. E.
Published: December 2017

Rates of proteolysis by hearts obtained from alloxan diabetic rats and perfused as working preparations with buffer simulating control sera were accelerated 30% above identically perfused control hearts. The total homogenate activities of cathepsin D and beta-N-acetylglucosaminidase, assayed in the presence of Triton X-100, decreased 15-35% in diabetic heart, but the activities of these lysosomal enzymes assayable in the absence of detergent were unchanged in the diabetic tissue. The effects of diabetes were examined further by centrifugation of particulate fractions from subtilisin-treated hearts of control and diabetic rats on polyvinylpyrrolidone-coated colloidal silica (Percoll) gradients. Two species of lysosomes were resolved on the basis of their densities. Both dense and buoyant lysosomes accumulated radioactivity when hearts were exposed to [14C]phenylalanine methyl ester. Dense lysosomes (1.06–1.09 g/ml) sedimented with mitochondria while buoyant lysosomes banded with Golgi and sarcolemmal particles (1.02–1.03 g/ml). When particulate fractions of hearts from diabetic animals were layered on the Percoll gradients, total activities of beta-N-acetylglucosaminidase and cathepsin D were decreased from control in buoyant lysosomes, but unchanged in dense lysosomes. These results demonstrated that the increase in proteolysis in the diabetic heart was associated with decreased total activity and latency of cathepsin D and beta-N-acetylglucosaminidase and an increased proportion of dense lysosomes in the particulate fraction.
Observations of 23Na in frog skin by NMR
Authors: Civan M. M. , Degani H. , Margalit Y. , Shporer M.
Published: December 2017

The shift reagent dysprosium tripolyphosphate [Dy(PPPi)7-2] has been applied to the nuclear magnetic resonance (NMR) study of 23Na in frog skin. The anion complex produces slight increases in the transcellular short-circuit current and conductance of the tissue. However, the electrophysiological responsiveness of the tissue to vasopressin, amiloride, and transient removal of extracellular potassium appear unimpaired. A large signal of [31P]phosphocreatine was also observed, further documenting that relatively little damage was produced by the reagent. Three signals of 23Na were observed, reflecting the sodium pools in an external standard solution, the extracellular space, and a medium relatively inaccessible to Dy(PPPi)7-2. The size of the reagent-insensitive signal could be increased by ouabain, a selective inhibitor of sodium extrusion, and could be decreased by amiloride, a selective inhibitor of sodium entry. The results indicate that the anion complex-insensitive signal reflects, at least in part, intracellular sodium and suggest that 23Na may become a useful complementary tool for monitoring the intracellular sodium content of transporting epithelia.
Ca2+, cAMP, and changes in myosin phosphorylation during contraction of smooth muscle
Authors: Aksoy M. O. , Mras S. , Kamm K. E. , Murphy R. A.
Published: December 2017

Phosphorylation of myosin increases rapidly upon stimulation of an arterial smooth muscle. However, peak values are not maintained and phosphorylation declines, while active stress increases monotonically to a sustained steady state. The aim of this study was to determine the reason(s) for the transient change in myosin phosphorylation. Four hypotheses were considered: 1) reduced substrate, i.e., ATP depletion, 2) altered access of either the myosin kinase or phosphatase to the cross bridge, 3) reduced myosin kinase activity secondary to its phosphorylation by adenosine 3',5'-cyclic monophosphate-dependent protein kinase, and 4) reduced myoplasmic [Ca2+] during the contraction. Our results suggest that the most likely explanation is that there are two Ca2+-dependent regulatory processes: 1) myosin phosphorylation and 2) a second, unidentified site allowing stress maintenance with reduced cross-bridge cycling rates. A higher cell Ca2+ concentration appears to be necessary to activate myosin kinase and produce myosin phosphorylation than is needed for force maintenance. We suggest that agonist-induced Ca2+ transients, coupled with the differential Ca2+ sensitivity of the two regulatory systems, may explain the observed transient in myosin phosphorylation during a maintained contraction in smooth muscle.
Alteration of luminal membrane structure by antidiuretic hormone
Authors: Hays R. M.
Published: December 2017

The final step in the action of antidiuretic hormone (ADH) is the insertion of aggregates of water-conducting particles into the luminal membrane of the receptor cell. The aggregates arise from cytoplasmic tubular structures that fuse with the luminal membrane. This review presents a number of questions about this process, along with current attempts to answer them. The following topics are addressed: 1) the exact role of the cytoskeleton in promoting tubular fusion, 2) the nature of the translocation process leading to fusion, 3) the point in the sequence at which ADH enters, 4) the composition and structure of the water-conducting particles. Some of the answers require a more complete understanding of the structure of the apical portion of ADH-sensitive cells; recent morphological studies are therefore included.
Calcium elevation in cultured heart cells: its role in cell injury
Authors: Murphy E. , Aiton J. F. , Horres C. R. , Lieberman M.
Published: December 2017

Inhibition of the Na+-K+ pump in cultured embryonic chick heart cells promotes the elevation of intracellular Ca2+ and is a useful manipulation to study the relationship between Ca2+ and myocardial cell injury. One hour of Na+-K+ pump inhibition resulted in a fourfold increase in cell Na+, a 50% decline in cell K+, and a 5- to 10-fold increase in cell Ca2+, 45% of which is mitochondrial. The degree of cell injury induced by Ca2+ loading was evaluated by monitoring the content of adenosine 5'-triphosphate (ATP) and the release of the intracellular enzyme lactate dehydrogenase (LDH). Under these conditions ATP content declined by 25-30% and LDH release increased from 1 to 1.4% of the total LDH. Furthermore, cells subjected to 1 h of Na+-K+ pump inhibition and returned to control solution for 5 h showed that Ca2+ decreased to near control levels and ATP content was restored. Although inhibition of Na+-K+ transport caused a large increase in cell Ca2+, neither Na+-K+ pump inhibition nor elevation in total cell Ca2+ per se resulted in irreversible myocardial cell injury.
Intracellularly recorded electrical activity of smooth muscle of guinea pig oviduct
Authors: Parkington H. C.
Published: December 2017

Action potentials of the simple spike form, similar to those observed in the longitudinal myometrium, were recorded intracellularly from the strands of longitudinal muscle that are associated with the guinea pig oviduct. Action potentials recorded from the circular layer were dominated by a prolonged (1- to 10-s) plateau component that was sometimes preceded by a spike. Electrotonic potentials in response to hyperpolarizing current pulses are consistent with the view that the plateau component might be associated with an increase in membrane conductance. Action potentials of circular smooth muscle were resistant to changes in the extracellular cationic concentration. Increasing external Ca2+ prolonged the duration of the plateau component of the action potential in the region nearest the uterus on days 3 and 4 after ovulation: Ca2+ had no consistent effect at other times during the estrous cycle or on ampullary segments at any time. Reducing the concentration of external Cl- in the presence of Na+ resulted in a significant increase in the amplitude of the plateau.