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Biomedical subjects

S S Miller

Publications and source records attributed to S S Miller.

At least 37 records · Page 2Linked to original sources
PubMed1997

Resident learning and knowledge retention from resident-prepared chest radiology conferences.

RATIONALE AND OBJECTIVES: The authors assessed resident learning and retention of knowledge from resident-prepared chest radiology conferences. MATERIALS AND METHODS: Radiology residents presented five chest conferences to their peers during a 5-month interval; the conferences were modeled on a case presentation format. Tests were given 5 minutes before each conference (pretest) and immediately after each conference (posttest). The tests were readministered as a final examination 6 months later, at which time the residents were asked to evaluate the conference format. RESULTS: Conference attendance ranged from six to 11 residents. Mean posttest scores were statistically significantly higher than mean pretest scores (P < .0001). Six-month retention scores were higher than pretest scores (P < .05) but lower than posttest scores (P < .05). On a scale of 1-6, with 1 representing strongly disagree and 6 strongly agree, residents strongly agreed that the conferences provided an excellent learning experience (mean score, 5.27). CONCLUSION: Resident-prepared conferences are effective for teaching residents chest radiology. Resident testing at 6 months demonstrated retention of knowledge above pretest levels but lower than posttest levels.

Analysis of Variance↗
PubMed1997

Nitrogen assimilation in alfalfa: isolation and characterization of an asparagine synthetase gene showing enhanced expression in root nodules and dark-adapted leaves.

Asparagine, the primary assimilation product from N2 fixation in temperate legumes and the predominant nitrogen transport product in many plant species, is synthesized via asparagine synthetase (AS; EC 6.3.5.4). Here, we report the isolation and characterization of a cDNA and a gene encoding the nodule-enhanced form of AS from alfalfa. The AS gene is comprised of 13 exons separated by 12 introns. The 5' flanking region of the AS gene confers nodule-enhanced reporter gene activity in transformed alfalfa. This region also confers enhanced reporter gene activity in dark-treated leaves. These results indicate that the 5' upstream region of the AS gene contains elements that affect expression in root nodules and leaves. Both AS mRNA and enzyme activity increased approximately 10- to 20-fold during the development of effective nodules. Ineffective nodules have strikingly reduced amounts of AS transcript. Alfalfa leaves have quite low levels of AS mRNA and protein; however, exposure to darkness resulted in a considerable increase in both. In situ hybridization with effective nodules and beta-glucuronidase staining of nodules from transgenic plants showed that AS is expressed in both infected and uninfected cells of the nodule symbiotic zone and in the nodule parenchyma. RNA gel blot analysis and in situ hybridization results are consistent with the hypothesis that initial AS expression in nodules is independent of nitrogenase activity.

Adaptation, Physiological↗
PubMed1997

Apical and basolateral membrane mechanisms that regulate pHi in bovine retinal pigment epithelium.

pH regulation was studied in fresh explant bovine retinal pigment epithelium-choroid using the pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and intracellular microelectrodes. Acid recovery was HCO3 dependent, inhibited by apical amiloride and apical or basal 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), and required apical and basal Na. Alkali recovery was HCO3 dependent and inhibitable by apical or basal DIDS. Three apical and two basolateral transporters were identified. Four contribute to acid extrusion, i.e., apical Na/H exchange, apical H-lactate cotransport, and apical Na-HCO3 cotransport and basolateral Na-HCO3 cotransport. At least two contribute to alkali extrusion, i.e., apical Na-HCO3 cotransport and a basolateral HCO3-dependent, DIDS-inhibitable mechanism, possibly Na-HCO3 cotransport, Cl/HCO3 exchange, or both. The apical Na-HCO3 cotransporter is electrogenic, carrying net negative charge inward. Basal Cl removal or addition of basal HCO3 caused HCO3- and Cl-dependent alkalinizations, respectively. Apical DIDS increased both responses. These cytosolic pH (pHi) regulatory mechanisms are so tightly coupled that changes in pHi can only occur after two or more of them are inhibited. In addition, these mechanisms help provide pathways for transport of Na and HCO3 across the retinal pigment epithelium between the blood and the distal retina.

Acids↗
PubMed1996

A new reusable instrument designed for simple and secure knot tying in laparoscopic surgery.

This new instrument is a metal cannula designed for use in creating controlled intracorporeal knots similar to instrument ties used in open surgery. It can therefore be used in a number of situations in laparoscopic surgery for ligation in continuity-for example, of the undivided cystic duct or of a vascular pedicle or for ligation of the cut end of a pedicle held in artery forceps. The cannula is designed to be loaded with 0 suture material and has a tip that readily penetrates the abdominal wall through a 3-mm skin incision, so it can be introduced either directly or through an introducer sleeve in a laparoscopic port. A curved dissecting forceps is used for knot formation. The knot is tightened by pushing the cannula with the other hand, and further throws are applied. The tip is profiled so that the knot, which is formed is pushed down into position without slippage or damage to the suture material.

Catheterization↗
PubMed1996

The anterior abdominal wall in laparoscopic procedures and limitations of laparoscopic simulators.

BACKGROUND: Maneuvers involving two-handed techniques such as intracorporeal suturing and knot-tying during laparoscopy are intrinsically more difficult than ones performed during open surgery. METHODS: The use of simulators to practice and teach specialized techniques is established. However, simulators vary greatly, and few, as yet, represent the abdominal wall well. RESULTS: This study has shown that the working angle between instruments during laparoscopic cholecystectomy is 78 degrees, and this can be increased to 117 degrees by moving the instruments laterally. In contrast, the working angle in four trainers assessed was never greater than 77 degrees. CONCLUSIONS: This suggests that some maneuvers may be more difficult in trainers than at surgery. This has implications for training and the further development of more realistic simulators.

Abdominal Muscles↗
PubMed1996

Cyclic responding by pigeons on the peak timing procedure.

The present experiment examined whether discrimination learning shapes the single-peaked response distributions usually obtained with the peak procedure. Two sources of learning in pigeons were disclosed: learning to respond near the time of reinforcement on fixed interval (FI) trials and learning to withhold responding once the FI duration had elapsed on peak interval (PI) trials. Pigeons also produced a highly unexpected second peak in responding on nonreinforced PI trials at 3 times the FI duration. Follow-up experiments showed that a 1:4 FI:PI duration ratio supported double peaks, but only 1 peak was obtained with a 1:8 FI:PI duration ratio. Finally, 4 peaks could be observed on extra-long PI trials under a 1:4:8 FI:PI:PI ratio procedure. The multiple-peaked response distributions are an unprecedented finding that present a major challenge to any theory of time perception.

Animals↗
PubMed1996

Nonsteroidal anti-inflammatory drugs alter chloride and fluid transport in bovine retinal pigment epithelium.

Nonsteroidal anti-inflammatory drugs (NSAIDs) were added to the solutions bathing the apical membrane of bovine retinal pigment epithelium (RPE)-choroid explants. For example, niflumic acid (100 microM) depolarized the basolateral membrane voltage (VB) by approximately 12 mV, increased transepithelial potential by 4.5 mV, decreased intracellular Cl activity by 13 mM, decreased transepithelial resistance by 17 omega.cm2, and increased the ratio of apical to basolateral membrane resistance nearly threefold. All of these changes are consistent with an increase in basolateral membrane Cl conductance. In addition, niflumic acid caused intracellular Ca concentration to decrease by 16 nM and fluid transport rate to increase by 1.5 microliters.cm-2.h-1. Flufenamic acid, which is structurally very similar to niflumic acid, had the opposite effects on membrane voltage and resistance. Basal application of the Cl channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid or current clamping VB to the reversal potential for Cl practically abolished the niflumic acid response. The niflumic acid results suggest that certain NSAIDs can directly alter Cl conductance in the bovine RPE, apparently independently of cyclooxygenase inhibition.

Animals↗
PubMed1995

The delayed basolateral membrane hyperpolarization of the bovine retinal pigment epithelium: mechanism of generation.

1. Conventional and ion-selective double-barrelled microelectrodes were used in an in vitro preparation of bovine retinal pigment epithelium (RPE)-choroid to measure the changes in membrane voltage, resistance and intracellular Cl- activity (aCli) produced by small, physiological changes in extracellular potassium concentration ([K+]o). These apical [K+]o changes approximate those produced in the extracellular (subretinal) space between the photoreceptors and the RPE following transitions between light and dark. 2. Changing apical [K+]o from 5 to 2 mM in vitro elicited membrane voltage responses with three distinct phases. The first phase was generated by an apical membrane hyperpolarization, followed by a (delayed) basolateral membrane hyperpolarization (DBMH); the third phase was an apical membrane depolarization. The present experiments focus on the membrane and cellular mechanisms that generate phase 2 of the response, the DBMH. 3. The DBMH was abolished in the presence of apical bumetanide (100 microM); this response was completely restored after bumetanide removal. 4. Reducing apical [K+]o, adding apical bumetanide (500 mM), or removing apical Cl- decreased aCli by 25 +/- 6 (n = 8), 28 +/- 1 (n = 2) and 26 +/- 5 mM (n = 3), respectively; adding 100 microM apical bumetanide decreased aCli by 12 +/- 2 mM (n = 3). Adding apical bumetanide or removing apical bath Cl- hyperpolarized the basolateral membrane and decreased the apparent basolateral membrane conductance (GB). 5. DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid) blocked the RPE basolateral membrane Cl- conductance and inhibited the DBMH and the basolateral membrane hyperpolarization produced by apical bumetanide addition or by removal of apical Cl-o. The present results show that the DBMH is caused by delta[K]o-induced inhibition of the apical membrane Na(+)-K(+)-2Cl- cotransporter; the subsequent decrease in aCli generated a hyperpolarization at the basolateral membrane Cl- channel.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1995

Alfalfa NADH-dependent glutamate synthase: structure of the gene and importance in symbiotic N2 fixation.

Glutamate synthase (GOGAT), a key enzyme in ammonia (NH+4) assimilation, occurs as two forms in plants: a ferredoxin-dependent form (Fd-GOGAT) and an NADH-dependent form (NADH-GOGAT). These enzymes are encoded by distinct genes as evidenced by their cDNA and deduced amino acid sequences. This paper reports the isolation and characterization of a NADH-GOGAT gene from alfalfa (Medicago sativa L.), the first GOGAT gene to be isolated from a eukaryote. RNase protection and primer extension experiments map the transcription start site of NADH-GOGAT to nearly identical positions. The transcribed region of this gene, 12,214 bp, is comprised of 22 exons separated by 21 introns. The 2.7 kbp region 5' from the translation initiation site confers nodule-specific reporter gene activity when used in a chimeric beta-glucuronidase (GUS) construct and transformed into Lotus corniculatus and Medicago sativa. Both infected and uninfected cells display GUS activity. The abundance of NADH-GOGAT transcripts increases substantially in developing nodules of plants infected with effective rhizobia. However, this increase is not observed when nodules are induced by a variety of ineffective rhizobial strains. Thus, unlike many other plant genes involved in root nodule NH+4 assimilation, high levels of NADH-GOGAT expression are strictly associated with effective nodules indicating that NADH-GOGAT plays a central role in the functioning of effective root nodules. An alfalfa Fd-GOGAT PCR product showing greater than 85% identity to maize Fd-GOGAT was isolated and used to investigate the contribution of this enzyme to NH+4 assimilation in nodules. Fd-GOGAT mRNA was abundant in leaves and cotyledons but was not detected in alfalfa root nodules. Fd-GOGAT in alfalfa does not appear to play a significant role in symbiotic N2 fixation.

Amino Acid Sequence↗
PubMed1995

Identification and functional characterization of a dual GABA/taurine transporter in the bullfrog retinal pigment epithelium.

Intracellular microelectrodes, fluorescence imaging, and radiotracer flux techniques were used to investigate the physiological response of the retinal pigment epithelium (RPE) to the major retinal inhibitory neurotransmitter, gamma-aminobutyric acid (GABA). GABA is released tonically in the dark by amphibian horizontal cells, but is not taken up by the nearby Müller cells. Addition of GABA to the apical bath produced voltage responses in the bullfrog RPE that were not blocked nor mimicked by any of the major GABA-receptor antagonists or agonists. Nipecotic acid, a substrate for GABA transport, inhibited the voltage effects of GABA. GABA and nipecotic acid also inhibited the voltage effects of taurine, suggesting that the previously characterized beta-alanine sensitive taurine carrier also takes up GABA. The voltage responses of GABA, taurine, nipecotic acid, and beta-alanine all showed first-order saturable kinetics with the following Km's: GABA (Km = 160 microM), beta-alanine (Km = 250 microM), nipecotic acid (Km = 420 microM), and taurine (Km = 850 microM). This low affinity GABA transporter is dependent on external Na, partially dependent on external Cl, and is stimulated in low [K]o, which approximates subretinal space [K]o during light onset. Apical GABA also produced a significant conductance increase at the basolateral membrane. These GABA-induced conductance changes were blocked by basal Ba2+, suggesting that GABA decreased basolateral membrane K conductance. In addition, the apical membrane Na/K ATPase was stimulated in the presence of GABA. A model for the interaction between the GABA transporter, the Na/K ATPase, and the basolateral membrane K conductance accounts for the electrical effects of GABA. Net apical-to-basal flux of [3H]-GABA was also observed in radioactive flux experiments. The present study shows that a high capacity GABA uptake mechanism with unique pharmacological properties is located at the RPE apical membrane and could play an important role in the removal of GABA from the subretinal space (SRS). This transporter could also coordinate the activities of GABA and taurine in the SRS after transitions between light and dark.

Animals↗
PubMed1995

Intravenous cholangiography and operative cholangiography: implications for laparoscopic cholecystectomy.

There has been a resurgence of interest in the potential role of intravenous cholangiography with the advent of laparoscopic cholecystectomy. A retrospective review of a historical group of 185 patients undergoing cholecystectomy in whom the results of both intravenous cholangiography (IVC) and routine operative cholangiography (OC) were available was carried out. The common bile duct (CBD) was explored in 31 (16.7%) patients and choledocholithiasis confirmed in 25 (81%). IVC had shown calculi in 17 of these 25 patients (sensitivity, 68%) whereas OC demonstrated calculi in 24 (sensitivity 96%). Out of the six negative duct explorations, IVC suggested stones in two patients (specificity 66%) and OC in four patients (specificity 33%). Ultrasound scan had a sensitivity of only 48% for bile duct calculi. The accuracy of IVC was 68% and OC 84%. Intravenous cholangiography has no routine role in the preoperative assessment in patients undergoing elective cholecystectomy. In high-risk patients, alternative imaging techniques should be used.

Case-Control Studies↗
PubMed1994

K+ and Cl- transport mechanisms in bovine pigment epithelium that could modulate subretinal space volume and composition.

1. Conventional and ion-selective double-barrelled microelectrodes were used in an in vitro bovine retinal pigment epithelium (RPE)-choroid preparation to measure the changes in membrane voltage, resistance and intracellular K+ and Cl- activities produced by small, physiological changes in extracellular potassium ([K+]o). 2. In the intact eye, light-induced changes in [K+]o occur in the extracellular (or subretinal) space that separates the neural retina and the RPE apical membrane. These [K+]o changes can be approximated in vitro by decreasing apical bath [K+]o from 5 to 2 mM. 3. This in vitro change in [K+]o simultaneously decreased intracellular Cl- and K+ activities (aCli and aKi) by 25 +/- 6 mM (n = 8) and 19 +/- 7 mM (n = 4) (mean +/- S.D.), respectively. In control Ringer solution (5 mM [K+]o) aCli and aKi were 65 +/- 10 mM (n = 28) and 65 +/- 8 mM (n = 6), respectively. 4. The [K+]o-induced decreases in aCli and aKi were both significantly inhibited, either by blocking the apical membrane K+ conductance with Ba2+ or the basolateral membrane Cl- conductance with DIDS (4,4'-diisothiocyano-stilbene-2,2'-disulphonic acid). 5. Transepithelial current pulses were used to determine the relative basolateral membrane Cl- conductance, TClBAS, was approximately 0.6 (n = 3), and the relative apical membrane K+ conductance, TKAP, was approximately 0.7 (n = 2). Step changes in basal bath [K+]o were used to estimate the relative basolateral membrane K+ conductance, TKBAS, was approximately 0.34 (n = 3). 6. These data show that the apical membrane K+ conductance and the basolateral membrane Cl- conductance are electrically coupled. In vivo, this coupling could have significant functional importance by modulating the relative hydration of the subretinal space, regulating RPE cell volume, and buffering the chemical composition of the subretinal space.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1994

Proton-lactate cotransport in the apical membrane of frog retinal pigment epithelium.

We studied lactate- and pyruvate-dependent proton transport across the apical membrane of frog RPE. The epithelium was mounted in a modified Ussing-chamber that allowed measurement of transepithelial potential and resistance while intracellular pH was measured with either intracellular microelectrodes or a pH-sensitive dye, 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF). To estimate the rate of lactate influx from the change in intracellular pH, we used the NH4 pulse technique to measure intracellular buffering capacity and its dependence on intracellular pH. We found that the buffering capacity was 16 mM at pH1 = 7.28, and that it increased as intracellular pH decreased. Intracellular pH was monitored with the tissue bathed in nominally HCO3-free (Hepes buffered) Ringer. The perfusate on the apical side of the epithelium was then changed to a Ringer that contained between 5 and 100 mM lactate or pyruvate. When 10-100 mM lactate or pyruvate was added to the apical bath the cells acidified by 0.05-0.50 pH units. For each of these acidifications, the initial acid influx into the RPE cells was calculated from the intracellular buffering capacity and the initial rate of intracellular acidification. These influxes were plotted as functions of the concentrations of lactate or pyruvate and this relationship was analysed using Michaelis-Menten kinetics. The Km values were: 33 +/- 5 mM for lactate and 9 +/- 3 mM for pyruvate. There were no differences in the rates of acid influx caused by L- or D-lactate. The rates of acidification caused by 50 mM apical L-lactate were reversibly reduced by 56% after apical administration of probenecid (2 mM), and irreversibly reduced by 63% after apical administration of the SH-reagent mersalyl acid (2 mM). These results indicate the presence of a proton-lactate cotransport system in the apical membrane of the frog RPE.

Animals↗
PubMed1994

Genomic structure, expression and evolution of the alfalfa aspartate aminotransferase genes.

Genomic clones encoding two isozymes of aspartate aminotransferase (AAT) were isolated from an alfalfa genomic library and their DNA sequences were determined. The AAT1 gene contains 12 exons that encode a cytosolic protein expressed at similar levels in roots, stems and nodules. In nodules, the amount of AAT1 mRNA was similar at all stages of development, and was slightly reduced in nodules incapable of fixing nitrogen. The AAT1 mRNA is polyadenylated at multiple sites differing by more than 250 bp. The AAT2 gene contains 11 exons, with 5 introns located in positions identical to those found in animal AAT genes, and encodes a plastid-localized isozyme. The AAT2 mRNA is polyadenylated at a very limited range of sites. The transit peptide of AAT2 is encoded by the first two and part of the third exon. AAT2 mRNA is much more abundant in nodules than in other organs, and increases dramatically during the course of nodule development. Unlike AAT1, expression of AAT2 is significantly reduced in nodules incapable of fixing nitrogen. Phylogenetic analysis of deduced AAT proteins revealed 4 separate but related groups of AAT proteins; the animal cytosolic AATs, the plant cytosolic AATs, the plant plastid AATs, and the mitochondrial AATs.

Amino Acid Sequence↗