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Biomedical subjects

R L Hudson

Publications and source records attributed to R L Hudson.

At least 19 recordsLinked to original sources

Effect of MT1 melatonin receptor deletion on melatonin-mediated phase shift of circadian rhythms in the C57BL/6 mouse.

In the mouse suprachiasmatic nucleus (SCN), melatonin activates MT1 and MT2 G-protein coupled receptors, which are involved primarily in inhibition of neuronal firing and phase shift of circadian rhythms. This study investigated the ability of melatonin to phase shift circadian rhythms in wild type (WT) and MT1 melatonin receptor knockout (KO) C57BL/6 mice. In WT mice, melatonin (90 microg/mouse, s.c.) administered at circadian time 10 (CT10; CT12 onset of activity) significantly phase advanced the onset of the circadian activity rhythm (0.60 +/- 0.09 hr, n = 41) when compared with vehicle treated controls (-0.02 +/- 0.07 hr, n = 28) (P < 0.001). In contrast, C57 MT1KO mice treated with melatonin did not phase shift circadian activity rhythms (-0.10 +/- 0.12 hr, n = 42) when compared with vehicle treated mice (-0.12 +/- 0.07 hr, n = 43). Similarly, in the C57 MT1KO mouse melatonin did not accelerate re-entrainment to a new dark onset after an abrupt advance of the dark cycle. In contrast, melatonin (3 and 10 pm) significantly phase advanced circadian rhythm of neuronal firing in SCN brain slices independent of genotype with an identical maximal shift at 10 pm (C57 WT: 3.61 +/- 0.38 hr, n = 3; C57 MT(1)KO: 3.45 +/- 0.11 hr, n = 4). Taken together, these results suggest that melatonin-mediated phase advances of circadian rhythms of neuronal firing in the SCN in vitro may involve activation of the MT2 receptor while in vivo activation of the MT1 and possibly the MT2 receptor may be necessary for the expression of melatonin-mediated phase shifts of overt circadian activity rhythms.

Animals↗

Mid- and far-infrared spectroscopic studies of the influence of temperature, ultraviolet photolysis and ion irradiation on cosmic-type ices.

Infrared (IR) studies of laboratory ices can provide information on the evolution of cosmic-type ices as a function of different simulated space environments involving thermal, ultraviolet (UV), or ion processing. Laboratory radiation experiments can lead to the formation of complex organic molecules. However, because of our lack of knowledge about UV photon and ion fluxes, and exposure lifetimes, it is not certain how well our simulations represent space conditions. Appropriate laboratory experiments are also limited by the absence of knowledge about the composition, density, and temperature of ices in different regions of space. Our current understanding of expected doses due to UV photons and cosmic rays is summarized here, along with an inventory of condensed-phase molecules identified on outer solar system surfaces, comets and interstellar grains. Far-IR spectra of thermally cycled H2O are discussed since these results reflect the dramatic difference between the amorphous and crystalline phases of H2O ice, the most dominant condensed-phase molecule in cosmic ices. A comparison of mid-IR spectra of products in proton-irradiated and UV-photolyzed ices shows that few differences are observed for these two forms of processing for the simple binary mixtures studied to date. IR identification of radiation products and experiments to determine production rates of new molecules in ices during processing are discussed. A new technique for measuring intrinsic IR band strengths of several unstable molecules is presented. An example of our laboratory results applied to Europa observations is included.

Carbonic Acid↗

A quick screening test of competency to stand trial for defendants with mental retardation.

19 terms and concepts from evaluations of competency to stand trial of 55 defendants with mental retardation were rated to examine whether a quick screening test could be devised that would differentiate those who were judged competent or not competent. A multiple regression and discriminant analysis gave four items that yielded maximum predictability (R = .84): court strategy, plead, testify, and jury. Guilty, trial, and prosecutor were also significantly more difficult for those who were not competent than those who were.

Adolescent↗

Bafilomycin-sensitive acid secretion by mantle epithelium of the freshwater clam, Unio complanatus.

This study reports the effects of a series of putative inhibitors of proton translocation on electrogenic acid secretion by the isolated shell-facing mantle epithelium of the freshwater clam, Unio complanatus. In this epithelium, adenosine 3',5'-cyclic monophosphate (cAMP)-dependent acid secretion is electrogenic and equivalent to the cAMP-dependent short-circuit current (Isc) when mounted in a Ussing-type flux chamber. Bafilomycin A1, a specific inhibitor of vacuolar-type H(+)-ATPases (V-type ATPases), inhibited cAMP-dependent electrogenic acid secretion, with an IC50 of 7 nM and a maximum effect at 100 nM. Inhibition of acid secretion by bafilomycin A1 occurred only when added to the shell side and was not reversed by repeated washings. Concomitant with the inhibition of cAMP-dependent Isc by bafilomycin A1, the electrical potential difference across the shell-facing membrane depolarized, and the conductance ratio of the apical to basolateral membranes decreased. The cAMP-dependent Isc and cAMP-dependent acid secretion also were inhibited by dicyclohexyl-carbodiimide, diethylstilbestrol, N-ethylmaleimide, and tributyltin. Amiloride, omeprazole, oligomycin, venturicidin, and ouabain had no effect on either cAMP-dependent Isc or cAMP-dependent acid secretion. The cAMP-dependent Isc and cAMP-dependent acid secretion were not affected by Na(+)-free or K(+)-free solutions. Finally, the cAMP-dependent Isc was strongly dependent on the pH of the shell-facing compartment, with 50% inhibition of the maximum cAMP-dependent Isc occurring at pH 6.0 and complete inhibition at pH 5.5.(ABSTRACT TRUNCATED AT 250 WORDS)

Acids↗

Ion transport by the isolated mantle epithelium of the freshwater clam, Unio complanatus.

Transepithelial ion transport was studied in three types of shell-facing mantle epithelia of the freshwater clam, Unio complanatus. These were 1) the epithelium isolated from near the margin of the shell; 2) the epithelium from the central region of a "normal" shell; and 3) the epithelium from the central region that was regenerating a damaged shell. Marginal and "shell-regenerating" epithelia produced a small (less than 5 mV) transepithelial voltage that was independent of PCO2 (0-4%). In the presence of 4% PCO2, the "normal" central epithelium produced a significantly larger transepithelial voltage (-8.5 +/- 0.99 mV), that, in addition, could be stimulated (-30.5 +/- 2.18 mV) by permeable analogues of adenosine 3',5'-cyclic monophosphate (cAMP) [e.g., 8-(4-chlorophenylthio)-adenosine 3',5'-cyclic monophosphate]. Voltage clamping the transepithelial voltage to zero resulted in a short-circuit current (Isc) that was consistent with positive charge moving from the hemolymph toward the shell. The measurements of transepithelial fluxes of 45Ca, 22Na, 42K, and 36Cl revealed that none of these ions, alone or in combination, could account for the stimulated Isc in "normal" mantle. In addition, similar measurements in marginal and "shell-regenerating" epithelia did not detect any significant net transepithelial flux of any of these ions. Acid extrusion at the shell-facing membrane and bicarbonate extrusion at the hemolymph-facing membrane were identified in "normal" central epithelium, but could not be found in marginal or "shell-regenerating" epithelia. These fluxes are equivalent to the cAMP-stimulated Isc in "normal" central epithelium.(ABSTRACT TRUNCATED AT 250 WORDS)

8-Bromo Cyclic Adenosine Monophosphate↗

Sodium-coupled glycine uptake by Ehrlich ascites tumor cells results in an increase in cell volume and plasma membrane channel activities.

The addition of 10 mM glycine to a physiological saline bathing Ehrlich ascites tumor cells is followed by a slow increase in cell volume that plateaus between 15 and 30 min at a level approximately equal to 17% greater than the control volume; this increase is not observed when glycine is added to cells suspended in a Na+-free saline. The results of studies using the patch-clamp technique in the cell-attached mode indicate that, 0.5-3 min after the addition of glycine to the bathing solution, there is a marked increase in the activity of single channels, which is almost all instances were previously present and operant in the plasma membrane. Successfully excised patches of membrane that contained a channel stimulated by glycine fell into two categories. Some became inactive within 15 sec in spite of the fact that the G omega seal remained intact. Others persisted for the lifetime of the seal. All of the persistent channels had an 11-fold selectivity for Cl- over K+ and a conductance of 23 pS when bathed by symmetrical 150 mM KCl solutions. Although the ionic specificities of the other channels have not been identified, there is reason to suspect that they might be K+ channels whose activities are dependent on factors lost when the patch is excised. Swelling induced by exposing these cells to a 50% hypotonic perfusate stimulated the activities of Cl- channels whose properties closely resemble those stimulated by the addition of glycine to the perfusate, strongly suggesting that the glycine-induced stimulation of Cl- channel activity is part of a volume-regulatory response to cell swelling. If the increase in channel activity induced by the addition of glycine to the perfusate is indeed a response to cell swelling, then this perfusate is indeed a response to cell swelling, then this volume-regulatory response must be extremely sensitive inasmuch as it appears to be "triggered" by an average increase in cell volume that does not exceed 5%.

Animals↗

Cell Na+ activities and transcellular Na+ absorption by descending colon from normal and Na+-deprived rabbits.

The relation between intracellular Na+ activities, (Na)c, determined employing Na+-selective microelectrodes, and the rates of active Na+ absorption, INa, by rabbit descending colon was examined when INa was varied over a wide range by chronic dietary Na+ deprivation. (Na)c averaged 13 mM and was independent of INa over a sixfold range. Further, the ratios of the slope resistance of the apical membrane (rm) to that of the basolateral membrane (rs) (i.e. rm/rs) in low-transporters (control diet) and high-transporters (Na+-deprived) did not differ significantly inspite of the fact that the Na+ conductance of the apical membranes of high-transporters was, on the average, three times greater than that of the low-transporters. These findings, together with the results reported by other laboratories, strongly suggest that the aldosterone-induced increase in the conductance of the apical membrane to Na+ and, in turn, the rate of entry of Na+ into the absorptive cells are followed by parallel increases in the ability of cells to extrude Na+ across the basolateral membrane in the absence of a sustained increase in (Na)c as well as the conductance of that barrier.

Animals↗

Effect of hypertonicity on the increase in basolateral conductance of Necturus small intestine in response to Na+-sugar cotransport.

Exposure of Necturus small intestine to a galactose-containing perfusate that is 20% hypertonic compared to the galactose-free (control) perfusate results in a rapid depolarization of the electrical potential difference across the apical membrane, psi mc, and a decrease in the ratio of the resistance of the apical membrane to that of the basolateral membrane, (rm/rs); however, the slow repolarization of psi mc and increase in (rm/rs), observed under isotonic conditions, is blocked. These findings are consistent with the notion that the increase in the conductance of the basolateral membrane in response to Na+-coupled sugar (or amino acid) transport across the apical membrane may be a 'volume regulatory response' to cell swelling.

Animals↗

Current-voltage relations of sodium-coupled sugar transport across the apical membrane of Necturus small intestine.

The current-voltage (I-V) relations of the rheogenic Na-sugar cotransport mechanism at the apical membrane of Necturus small intestine were determined from the relations between the electrical potential difference across the apical membrane, psi mc, and that across the entire epithelium, psi ms, when the latter was varied over the range +/- 200 mV, under steady conditions in the presence of galactose and after the current across the apical membrane carried by the cotransporter, ImSNa, is blocked by the addition of phloridzin to the mucosal solution. ImSNa was found to be strongly dependent upon psi mc over the range -50 mV less than psi mc less than EmSNa where EmSNa is the "zero current" or "reversal" potential. Over the range of values of psi mc encountered under physiological conditions the cotransporter may be modeled as a conductance in series with an electromotive force so that ImSNa = gmSNa (EmSNa - psi mc) where gmSNa is the contribution of this mechanism to the conductance of the apical membrane and is "near constant." In several instances ImSNa "saturated" at large hyperpolarizing or depolarizing values of psi mc. The values of EmSNa determined in the presence of 1, 5, and 15 mM galactose strongly suggest that if the Na-galactose cotransporters are kinetically homogeneous, the stoichiometry of this coupled process is unity. Finally, the shapes of the observed I-V relations are consistent with the predictions of a simple kinetic model which conforms with current notions regarding the mechanico-kinetic properties of this cotransport process.

Animals↗

Transcellular sodium transport and intracellular sodium activities in rabbit gallbladder.

This study was designed to explore the relation between the rate of transcellular active Na+ transport by rabbit gallbladder epithelium, JNa, and the intracellular Na+ activity, (Na)c; the latter was determined by use of highly selective Na+ microelectrodes. The underlying strategy was based on the well-established observation that JNa is stimulated by the presence of bicarbonate in the bathing solutions. Our results confirm previous observations that the addition of bicarbonate to the bathing solutions results in a twofold increase in JNa. In the absence of bicarbonate, (Na)c averaged 16 mM. Within 2-4 min after the addition of bicarbonate to both bathing solutions, (Na)c increased to an average value of 22 mM and then gradually declined and by 15 min did not differ significantly from the value observed in the absence of bicarbonate. Thus, a twofold increase in JNa is not associated with an increase in (Na)c. These results are in accord with earlier observations on Necturus urinary bladder and small intestine and contradict the notion that an increase in the rate of active Na+ extrusion from the cell across the basolateral membrane in response to an increase in the rate of Na+ entry across the apical membrane is necessarily the result of a sustained increase in (Na)c.

Animals↗

Kinetics of the effect of amiloride on the permeability of the apical membrane of rabbit descending colon to sodium.

The effects of the addition of graded concentrations of amiloride, (A)m, to the mucosal bathing solution on the permeability of the apical membrane of rabbit descending colon to Na (PmNa) were determined when the Na activity in the mucosal bathing solution, (Na)m, was 18, 32 or 100 mM. PmNa was obtained from current-voltage relations determined on tissues bathed with a high-K serosal solution before and after the addition of a maximally inhibitory concentration of amiloride to the mucosal solution as described by Turnheim et al. (Turnheim, K., Thompson, S.M., Schultz, S.G. 1983. J. Membrane Biol. 76:299-309). The results indicate that: (1) As demonstrated previously (Turnheim et al., 1983), PmNa decreases with increasing (Na)m. (2) PmNa also decreases hyperbolically with increasing (A)m. Kinetic analyses of the effect of amiloride on PmNa are consistent with the conclusions that: (i) the stoichiometry between the interaction of amiloride with apical membrane receptors that results in a decrease in PmNa is one-for-one; (ii) there is no evidence for cooperativity between amiloride and these binding sites; (iii) the value of (A)m needed to halve PmNa at a fixed (Na)m is 0.6-1.0 microM; and, (iv) this value is independent of (Na)m over the fivefold range studied. These findings are consistent with the notion that the sites with which amiloride interacts to bring about closure of the channels through which Na crosses the apical membrane are kinetically distinct from the sites with which (Na)m interacts to bring about closure (i.e., "self-inhibition"). In short, the effects of (Na)m and (A)m on PmNa in this tissue appear to be independent and additive.

Amiloride↗

Electrophysiological studies of sodium cotransport in epithelia: toward a cellular model.

During the past two decades, microelectrophysiological studies of small intestine and renal proximal tubule employing conventional as well as ion-selective microelectrodes have contributed significantly to our understanding of the nature of Na-coupled entry processes at the apical membrane as well as the overall workings of the simple model illustrated in FIGURE 1. These studies have unequivocally established the rheogenic and conductive nature of the Na-coupled sugar and amino-acid entry processes across the apical membrane of small intestine (and renal proximal tubule) and have, in addition, disclosed that the properties of the basolateral membrane respond to an increase in Na-coupled solute entry with an increase in the ability of the Na-K pump to extrude Na with little or no change in (Na)c32 and a parallel increase in the conductance of that barrier to K. Although these responses may be "triggered" by cell swelling, it is unclear how a cell "recognizes" minimal swelling and how this recognition, in turn, culminates in the observed changes in basolateral membrane pump-leak properties. Clearly, these findings have brought us to the interfaces between cell physiology and cell and molecular biology and have raised a number of intriguing questions that focus on the more global question: How do epithelial cells work?

Amino Acids↗

Sodium-coupled sugar transport: effects on intracellular sodium activities and sodium-pump activity.

Intracellular sodium activities, (Na)c, were determined in Necturus small intestine before and after addition of galactose to the mucosal bathing solution. In the absence of galactose, (Na)c averaged 12 millimoles per liter. Within 2 minutes after the addition of galactose to the mucosal solution, (Na)c increased to a mean value of 20 millimoles per liter and then declined, in parallel with an increase in transcellular sodium transport, to a value that did not differ significantly from that observed in the absence of the sugar. The final steady state in the presence of galactose was characterized by a three- to fourfold increase in the rate of transcellular Na+ transport in the absence of a significant increase in (Na)c. Thus, the increase in steady-state basolateral pump activity cannot be attributed to an increase in the intracellular sodium transport pool.

Animals↗

Cell swelling increases a barium-inhibitable potassium conductance in the basolateral membrane of Necturus small intestine.

Previous studies have shown that, immediately after the addition of galactose or alanine to the solution bathing the mucosal surface of Necturus small intestine, there is a rapid depolarization of the electrical potential difference across the mucosal membrane (psi mc). This is followed by a repolarization of psi mc that is paralleled by an increase in the ratio of the effective resistance of the mucosal membrane to that of the basolateral membrane (rm/rs); the latter was shown to be, at least in part, due to a marked increase in the conductance of the basolateral membrane. We now report the following. (i) Exposure of this epithelium to a 12% hypotonic solution results in a hyperpolarization of psi mc and an increase in rm/rs. These effects are blocked by metabolic inhibitors and by the presence of 5 mM Ba2+ in the bathing solution; indeed, in the presence of Ba2+, psi mc depolarizes and rm/rs decreases to low values. (ii) Addition of 15 mM galactose to the mucosal solution when the serosal solution alone contains 5 mM Ba2+ results in a depolarization of psi mc but no subsequent repolarization of psi mc or increase in rm/rs; however, psi mc repolarizes and rm/rs increases when Ba2+ is subsequently removed from the serosal bathing solution. We conclude that (i) the basolateral membrane normally possesses a Ba2+-inhibitable K conductance, which appears to be reduced in the presence of metabolic inhibitors; (ii) after exposure of the tissue to a hypotonic solution or the addition of galactose to the mucosal solution, this conductance increases; and (iii) these responses can be blocked by metabolic inhibitors. These findings suggest that the delayed response of this tissue to the addition of sugars or amino acids to the mucosal solution may be the result of cell swelling resulting from the intracellular accumulation of these solutes in osmotically active forms.

Amino Acids↗

Treatment of styes and meibomian cysts. Practical procedures.

Styes and meibomian cysts are common eye problems presenting to the general practitioner and both can be simply managed in the consulting rooms. Meibomian cysts, however, may present difficulties to the inexperienced and Dr Hudson has outlined a comprehensive plan of assessment and surgical management.

Cysts↗

Sickle cell anemia and transposition of the great vessels.

A child with homozygous sickle cell disease and transposition of the great vessels had erythrocytosis associated with markedly increased plasma erythropoietin activity. Her clinical course was complicated by neurologic manifestations but not by recurrent sickle cell vasooculsive episodes. The fetal hemoglobin level which had been greater than 25% during the first two years of life gradually decreased to less than 10%. She died at 3 years of age of congestive heart failure and severe anemia. The only sickle cell painful crisis occurred during her terminal illness. It is likely that the high levels of fetal hemorglobin decreased sickling and thus allowed erythrocytosis to develop. Fetal hemoglobin may also have prevented frequent vaso-occlusive events despite the high hematocrit level.

Anemia, Sickle Cell↗