Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 21
Filter
1.
Comput Methods Biomech Biomed Engin ; 10(2): 111-20, 2007 Apr.
Article in English | MEDLINE | ID: mdl-18651277

ABSTRACT

The Mennen femur plate is a fixation device used for the treatment of femoral periprosthetic fractures. It features a novel fastening method where curved prongs are plastically deformed securing the implant to the bone. Although this "clamp-on" method has been successfully used to treat fractures of long bones, there are no literature data assessing the nature of the required plastic deformation. In the present study, the parameters influencing the performance of the prongs were identified and further explored using numerical modeling. The new Mennen 3 PeriPro plate is briefly discussed focusing on the new sculpted formation of the prongs. Their design was optimized to effectively control the magnitude and position of the required plastic deformation achieving enhanced anchorage on the fractured bone with minimum effort. The work presented contains all the necessary steps in analysing a clinical problem using finite elements and illustrates how effective use of simulation techniques can accurately predict and effectively control the required plastic deformation of a structure.


Subject(s)
Bone Plates , Computer-Aided Design , Fracture Fixation, Internal/instrumentation , Models, Theoretical , Plastics/chemistry , Computer Simulation , Elasticity , Equipment Failure Analysis , Prosthesis Design , Stress, Mechanical
2.
Proc Inst Mech Eng H ; 220(7): 775-85, 2006 Oct.
Article in English | MEDLINE | ID: mdl-17117766

ABSTRACT

The Mennen femur plate is an internal fixation device used for the management of femoral perisprosthetic fractures, usually after total hip replacement surgery. The implant uses a number of curved prongs that embrace the fractured bone around its circumference without interfering with the stem of the prosthesis. Although the device has been used with considerable clinical success since its first introduction, a number of negative clinical results have been reported in the literature. The failure modes of the device are described and an evaluation of its performance is briefly presented. Based on this assessment as well as comments in the open literature, modifications in the design of the device have been implemented. The new Mennen 3 PeriPro plate is presented, with all the necessary data for a coherent explanation of its improved characteristics as defined using numerical simulations and experimental tests. The new device has all the beneficial features of the previous plate with improved structural performance and fatigue life and new sculpted formation of the prongs, providing a simple implantation technique with maximum gripping and minimum effort from the surgeon. The unique mode of fixation has been further improved, providing ample anchorage on the fracture bone without compromising its biomechanical integrity. By combining the device with a cable system, the spectrum of applications will be further expanded, enabling the surgeon to treat a broader range of fracture patterns.


Subject(s)
Bone Plates , Femoral Fractures/etiology , Femoral Fractures/surgery , Fracture Fixation, Internal/instrumentation , Hip Prosthesis/adverse effects , Computer Simulation , Elasticity , Equipment Failure Analysis , Finite Element Analysis , Fracture Fixation, Internal/methods , Humans , Models, Theoretical , Prosthesis Design , Stress, Mechanical , Tensile Strength
3.
J Neurochem ; 73(5): 1816-27, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10537039

ABSTRACT

Schwann cells cloned from rat sciatic nerve survive and display self-induced growth suppression, or undergo spontaneous apoptosis, on long-term serum-free subconfluent culture. Strain SCL4.1/F7 sustained the capacity to growth arrest for up to 40 generations. A soluble activity transmitted between neighbouring cells of this strain suppresses DNA synthesis within three cell cycles. Autocrine Schwann cell growth-inhibitory factor (SGIF) operates during the G1 phase of the cell cycle, overcomes the mitogenic action of Schwann cell/serum-associated (platelet-derived growth factor-BB) and axon-associated (axolemma-enriched fraction) stimuli in serum-free conditions, and suppresses DNA synthesis in sciatic nerve Schwann cell cultures in a stage-specific manner. A 35-kDa protein with N-terminal sequence and approximate molecular mass of the C-propeptide of rat alpha1-procollagen I makes a major contribution to SGIF. Growth suppression in the SCL4.1/F7 strain is mediated by the ras/extracellular signal-regulated kinase pathway, is accompanied by down-regulation of erbB2/erbB3 and of tetraethylammonium-sensitive K+ currents, and is followed by transition of cells within 5-10 days from O4+, p75 nerve growth factor receptor (p75NGF-R)+, glial fibrillary acidic protein (GFAP)+ to O4+, p75NGF-R-, GFAP-, periaxin+ phenotypes. Oct-6/SCIP mRNA is present in both proliferating and growth-arrested SCL4.1/F7 cells. These results demonstrate an autocrine/ paracrine loop for the growth arrest of clonally derived Schwann cells in the absence of axons linked in part to the metabolism of collagen. Schwann cells thus appear to self-regulate growth in a negative as well as a positive direction through characterized molecular mechanisms and signal pathways.


Subject(s)
Growth Inhibitors , Peptide Fragments/pharmacology , Procollagen/pharmacology , Schwann Cells/cytology , Animals , Becaplermin , Cell Differentiation/drug effects , Cell Division/drug effects , Cells, Cultured , Culture Media, Conditioned , DNA/biosynthesis , G1 Phase , Growth Inhibitors/analysis , Growth Inhibitors/pharmacology , Mitogens/pharmacology , Myelin Sheath/physiology , Platelet-Derived Growth Factor/pharmacology , Potassium Channels/physiology , Proto-Oncogene Proteins c-sis , Rats , Receptor Protein-Tyrosine Kinases/metabolism , Receptor, EphA8 , Receptor, ErbB-2/metabolism , Schwann Cells/drug effects , Sciatic Nerve/cytology , Stem Cells/cytology , Stem Cells/drug effects , Time Factors
4.
Glia ; 22(2): 113-20, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9537831

ABSTRACT

Mitogenesis in a variety of tissues is known to be inhibited by K+ channel blockers such as tetraethylammonium (TEA) and 4-aminopyridine (4-AP). Using radiolabeled thymidine as a proliferation index we have examined what role, if any, specific K+ channels have in cultured Schwann cells that have been induced to proliferate by pre-exposure to mitogens. TEA and 4-AP are "broad-spectrum" in that they block a variety of different types of K+ channel. In contrast, we found that alpha-dendrotoxin (alpha-DTX), a specific blocker of the type 1 fast delayed rectifier current (the largest component of Schwann cell K+ current) does not affect proliferation, suggesting that type 1 current may not be involved in mitogenesis. This suggestion is supported by our finding that the values of the KD for the mitogenic effect (722 nM, 4-AP; 13 mM, TEA) are much larger than the corresponding electrophysiological values for type 1 channels (0.1 mM, 4-AP; 0.2 mM, TEA). Charybdotoxin (200 nM) and iberiotoxin (100 nM), inhibitors of Ca2+-activated K+ channels, cesium (5 mM), an inhibitor of inward rectifier channels, and furosemide (100 pM), which blocks Na+/K+/Cl- cotransport, all had no effect on proliferation. Interestingly, 4,4'-diisothiocyanatostilbene 2,2'-disulphonate (DIDS), which blocks voltage-gated Cl- channels, reduced proliferation. In summary, broad-spectrum K+ channel blockers inhibit Schwann cell proliferation, but inhibitors specific for type 1, Ca2+-activated, and inward rectifier K+ channels do not. Whether the inhibition is mediated by type 2 K- channels, by an as yet unidentified Schwann cell K+ channel, or by another mechanism remains unclear.


Subject(s)
Ion Channels/antagonists & inhibitors , Ion Channels/metabolism , Schwann Cells/metabolism , 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/pharmacology , 4-Aminopyridine/pharmacology , Animals , Cell Division/physiology , Cells, Cultured , Elapid Venoms/pharmacology , Electrophysiology , Indicators and Reagents , Mitosis/drug effects , Mitosis/physiology , Neurotoxins/pharmacology , Peptides/pharmacology , Potassium Channels/drug effects , Potassium Channels/metabolism , Rabbits , Schwann Cells/drug effects , Sciatic Nerve/cytology , Sciatic Nerve/drug effects , Tetraethylammonium/pharmacology
5.
J Neurosci Res ; 44(2): 191-8, 1996 Apr 15.
Article in English | MEDLINE | ID: mdl-8723228

ABSTRACT

Astrocytes actively maintain their intracellular pH (pHi) more alkaline than expected by passive distribution of H+. Acid extruding transporters such as the amiloride-sensitive Na+/H+ exchanger (NHE) are necessary for pH regulation. Currently, four mammalian NHEs (NHE1-NHE4) have been cloned, with a fifth (NHE5) partially cloned. We attempted to determine which isoform(s) of NHE was present in cultured hippocampal astrocytes using amiloride sensitivity and immunospecificity as criteria. In the absence of HCO3-, amiloride blocked pHi recovery after an acid load with an IC50 of approximately 3.18 microM, similar to values reported for the amiloride-sensitive isoforms NHE1 and NHE2. Immunoblotting with a highly specific antibody for NHE1 identified a 100 kDa protein, indicating the presence of NHE1 in whole brain, hippocampus, and cultured hippocampal astrocytes. Further probing for an additional amiloride-sensitive NHE failed to detect evidence of the presence of NHE4. Surprisingly, application of the potent analog of amiloride, ethylisopropylamiloride (EIPA), caused a reversible alkalinization of pHi, suggesting the presence of an additional acid/base transport mechanism that is EIPA-sensitive.


Subject(s)
Astrocytes/metabolism , Hippocampus/metabolism , Sodium-Hydrogen Exchangers/metabolism , Amiloride/analogs & derivatives , Amiloride/pharmacology , Animals , Animals, Newborn , Astrocytes/drug effects , Bicarbonates/pharmacology , Cell Membrane/metabolism , Cells, Cultured , Gene Expression , Hydrogen-Ion Concentration , Rats , Rats, Sprague-Dawley , Sodium-Hydrogen Exchangers/biosynthesis , Sodium-Hydrogen Exchangers/drug effects
6.
Glia ; 16(1): 7-15, 1996 Jan.
Article in English | MEDLINE | ID: mdl-8787769

ABSTRACT

Octanol rapidly closes gap junction channels but its mechanism of action is not known. Because intracellular [H+], pHi, also affects the conductance of gap junctions, we studied octanol's effects on pHi in cultured rat astrocytes, which are highly coupled cells. Octanol (1 mM) caused an acid shift in the pHi of 90% of rat hippocampal astrocytes which averaged -0.19 +/- 0.09 pH units in magnitude. In 58% of the cells tested, a biphasic change in pHi was seen; octanol produced an initial acidification lasting approximately 10 min that was followed by a persistent alkalinization. The related gap junction uncoupling agent, heptanol, had similar effects on pHi. Octanol-induced changes in pHi were similar in nominally HCO(3-)-free and HCO(3-)-containing solutions, although the rate of initial acidification was significantly greater in the presence of HCO3-. The initial acidification was inhibited in the presence of the stilbene DIDS, an inhibitor of Na+/HCO3- cotransport, indicating that octanol caused acidification by blocking this powerful acid extruder. The alkalinization was inhibited by amiloride which blocks the Na+/H+ exchanger (NHE), an acid extruder, suggesting that the alkaline shift induced by octanol was caused by stimulation of NHE. As expected, octanol's effects on astrocytic pHi were prevented by removal of external Na+, which blocks both Na+/HCO3- cotransport and NHE. Octanol had only small effects on intracellular Ca2+ (Ca2+i) in astrocytes. Hepatocytes which, like astrocytes, are strongly coupled to one another, showed no change in pHi with octanol application. Fluorescence recovery after photobleaching (FRAP) was used to study the effect of changes in astrocyte pHi on degree of coupling in hippocampal astrocytes. Coupling was decreased by intracellular acid shifts approximately -0.2 pH units in size. Octanol's effects on astrocyte pHi were complex but a prompt initial acidification was nearly always seen and could contribute to the uncoupling action of this drug in astrocytes. Because octanol uncouples hepatocytes without changing their pHi, this compound clearly can influence gap junctional conductance independent of changes in pHi.


Subject(s)
Astrocytes/metabolism , Gap Junctions/drug effects , Octanols/pharmacology , Uncoupling Agents/pharmacology , 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/pharmacology , Amiloride/pharmacology , Animals , Animals, Newborn , Astrocytes/drug effects , Astrocytes/ultrastructure , Calcium/metabolism , Cells, Cultured , Diuretics/pharmacology , Gap Junctions/ultrastructure , Hippocampus/cytology , Hippocampus/drug effects , Hippocampus/metabolism , Hydrogen-Ion Concentration , Image Processing, Computer-Assisted , Microscopy, Video , Rats , Rats, Sprague-Dawley , Sodium/physiology
7.
Neuroreport ; 6(1): 193-6, 1994 Dec 30.
Article in English | MEDLINE | ID: mdl-7703413

ABSTRACT

Astrocyte proliferation was studied in primary cultures of rat spinal cord by [3H]thymidine uptake and was significantly reduced by culturing cells for 24 h in the presence of agents known to block astrocytic K+ channels: Cs+, Ba2+, 4-AP and tetraethylammonium (TEA). To determine whether effects were mediated by changes in Vm or pHi, these parameters were studied electrophysiologically or ratiometrically, using BCECF. Of the four K+ channel blockers, only Ba2+ depolarized astrocytes significantly. However, all four K+ channel blockers resulted in an alkaline shift in pHi. Under culture conditions that altered pHi in a defined way, proliferation strongly depended on pHi, with highest rates at pH approximately 6.7 and growth inhibition at more acidic or alkaline conditions. These observations suggest that astrocyte proliferation is sensitive to changes in pHi and that K+ channel blockers may exhibit their antiproliferative effects through changes in pHi.


Subject(s)
Hydrogen-Ion Concentration , Neuroglia/cytology , Neuroglia/physiology , Potassium Channel Blockers , Animals , Astrocytes/cytology , Astrocytes/physiology , Cell Division/physiology , Electrophysiology , Fluoresceins , Fluorescent Dyes , Rats , Rats, Sprague-Dawley
8.
J Neurophysiol ; 72(6): 2816-26, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7897491

ABSTRACT

1. Depolarization of glial cells causes their intracellular pH (pHi) to increase. To more completely characterize this depolarization-induced alkalinization (DIA) in mammalian astrocytes, we studied DIA in cultured rat hippocampal astrocytes. Astrocytes were loaded with the fluorescent pH indicator 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF), and pHi was monitored with the use of an imaging system. Cells were studied approximately 24 h after removing them from serum-containing culture medium. In HCO-3-buffered solution containing 3 mM K+, mean baseline pHi was 7.14 +/- 0.14 (mean +/- SD). 2. Astrocyte pHi rapidly and reversibly alkalinized when bath [K+] was increased from 3 to 12 mM. In HCO-3-buffered solution, mean DIA amplitude was 0.16 +/- 0.01 pH units, and mean rate of pHi change was 0.076 +/- 0.03 pH units/min. In contrast, DIA elicited in nominally HCO-3-free, N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered solution was much smaller (0.03 +/- 0.01 pH units, 0.04 +/- 0.01 pH units/min; P < 0.0001), indicating that DIA was, in large part, a HCO-3-dependent process. Subsequent experiments were carried out in HCO-3-buffered solution. 3. The relationship between DIA and variable changes in bath [K+] was examined. Increasing bath [K+] from 3 to 6 mM produced a DIA of 0.07 +/- 0.04 pH units, and lowering [K+] to 0.5 mM resulted in an acid shift of 0.08 +/- 0.05 pH units. The effects of these changes in [K+] on membrane potential were measured in separate experiments by whole cell patch-clamp recording. On the basis of these data, it was possible to construct a relationship between Vm and pHi; shifting membrane potential approximately 10 mV resulted in a pHi shift of approximately 0.07. 4. Application of 0.5 mM Ba2+ depolarized Vm and elicited DIA in astrocytes. This indicated that depolarization, in the absence of an increase in [K+], could cause DIA. Application of Ba2+ also blocked K(+)-induced DIA, presumably because blockade of K+ channels prevented any depolarization by K+. 5. Cells with more alkaline baseline pHis exhibited larger and more rapidly developing DIAs. The mechanism of this effect is not known. 6. The timing of serum removal affected astrocyte DIA. Cells studied approximately 24 h after serum removal always exhibited robust DIA (mean = 0.16 +/- 0.01 pH units).(ABSTRACT TRUNCATED AT 400 WORDS)


Subject(s)
Astrocytes/physiology , Hippocampus/physiology , 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/pharmacology , 4-Aminopyridine/pharmacology , Animals , Astrocytes/drug effects , Astrocytes/metabolism , Barium/pharmacology , Bicarbonates/metabolism , Bicarbonates/pharmacology , Biological Transport/drug effects , Biological Transport/physiology , Cells, Cultured , Chlorides/metabolism , Hippocampus/cytology , Hippocampus/metabolism , Hydrogen-Ion Concentration , Kinetics , Membrane Potentials/drug effects , Potassium/pharmacology , Rats , Rats, Sprague-Dawley , Sodium/metabolism , Sodium/physiology
9.
J Neurosci ; 14(5 Pt 1): 2464-75, 1994 May.
Article in English | MEDLINE | ID: mdl-8182422

ABSTRACT

Astrocytes in vitro and in situ have been shown to express voltage-activated ion channels previously thought to be restricted to excitable cells, including voltage-activated Na+, Ca2+, and K+ channels. However, unlike neurons, astrocytes do not generate action potentials, and the functional role of voltage-activated channels in astrocytes has been an enigma. In order to study the function of Na+ channels in glial cells, we carried out ion flux measurements, patch-clamp recordings, and ratiometric imaging of [Na+]i during blockade of Na+ channels on rat spinal cord astrocytes cultured for 7-10 d. Acute blockade of astrocyte Na+ channels by TTX had multiple effects: (1) TTX reduced, in a dose-dependent manner, Na+/K(+)-ATPase activity measured as unidirectional influx of 86Rb+; (2) TTX depolarized astrocyte membrane potential at a rate of approximately 1 mV/min; (3) TTX (100 microM) reduced [Na+]i; and (4) prolonged exposure to micromolar TTX induced astrocyte death. All these effects of TTX could be mimicked by ouabain or strophanthidin, specific blockers of the Na+/K(+)-ATPase. The effects of TTX and ouabain (or strophanthidin) were not additive. These results suggest that TTX-blockable Na+ channels in glial cells serve functions that do not require their participation in action potential electrogenesis; in particular, we propose that glial Na+ channels constitute a "return" pathway for Na+/K(+)-ATPase function, which permits Na+ ions to enter the cells to maintain [Na+]i at concentrations necessary for activity of the Na+/K(+)-ATPase. Since astrocyte Na+/K(+)-ATPase is believed to participate in [K+]o homeostasis in the CNS, the coupling of Na+ flux through voltage-activated Na+ channels to ATPase activity may provide a feedback loop that participates in the regulation of K+ ion levels in the extracellular space.


Subject(s)
Astrocytes/physiology , Ganglia, Spinal/physiology , Sodium Channels/physiology , Sodium-Potassium-Exchanging ATPase/metabolism , Tetrodotoxin/pharmacology , Animals , Animals, Newborn , Astrocytes/drug effects , Astrocytes/enzymology , Astrocytoma , Cell Line , Cells, Cultured , Electrophysiology/methods , Ganglia, Spinal/cytology , Glioma , Membrane Potentials/drug effects , Membrane Potentials/physiology , Models, Biological , Ouabain/pharmacology , Rats , Rats, Sprague-Dawley , Rubidium/metabolism , Sodium/metabolism , Sodium Channels/drug effects , Strophanthidin/pharmacology , Time Factors , Tumor Cells, Cultured
10.
Glia ; 9(4): 280-91, 1993 Dec.
Article in English | MEDLINE | ID: mdl-8112821

ABSTRACT

Relatively little is known about the mechanisms of pHi regulation in mammalian glial cells. We analyzed pHi regulation in rat hippocampal astrocytes in vitro using the pH-sensitive dye BCECF. All experiments were carried out in CO2/HCO3(-)-free solutions. Recovery from NH4(+)-induced acid loads was strongly dependent on the presence of extracellular Na+ and was inhibited by amiloride and its more specific analog EIPA, indicating the presence of Na(+)-H+ exchange in these cells. Removing bath Na+ or adding amiloride caused resting pHi to shift in the acid direction. Even in the absence of bath Na+ or presence of Na+/H+ inhibitors, however, these astrocytes continued to show significant recovery from acid loads. The mechanism of this amiloride-insensitive and Na(+)-independent pHi recovery process was sought and appeared to be a proton pump. In the absence of Na+, recovery from an acid load was completely blocked by the highly specific blocker of vacuolar-type (v-type) H+ ATPase, bafilomycin A1 (BA1). In normal Na+ containing solutions, exposure to BA1 caused a small acid shift in baseline pHi and slowed recovery rate from NH4(+)-induced acid loads by about 32%. The rate of Na(+)-independent pHi recovery was increased by depolarization with 50 mM [K+] solution, and this effect was rapidly reversible and blocked by BA1. These results indicate that, in CO2/HCO3(-)-free solution, pHi regulation in hippocampal astrocytes was mediated by Na(+)-H+ exchange and by a BA1-inhibitable proton pump. Because the proton pump's activity was influenced by membrane potential, this acid exporting mechanism could contribute to the depolarization-induced alkalinization that is seen in astrocytes. Although v-type H(+)-ATPase had been previously isolated from the brain, this is the first report indicating that it has a role in regulating pHi in brain cells.


Subject(s)
Anti-Bacterial Agents/pharmacology , Astrocytes/drug effects , Hippocampus/drug effects , Macrolides , Proton-Translocating ATPases/metabolism , Sodium-Hydrogen Exchangers/drug effects , Amiloride/pharmacology , Animals , Astrocytes/metabolism , Electric Stimulation , Fluoresceins/pharmacology , Hippocampus/cytology , Hydrochloric Acid/pharmacology , Hydrogen-Ion Concentration , Membrane Potentials/drug effects , Nerve Tissue Proteins/metabolism , Quaternary Ammonium Compounds/pharmacology , Rats , Rats, Sprague-Dawley
11.
Am J Physiol ; 263(6 Pt 2): F1011-9, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1282781

ABSTRACT

The effects of the beta-adrenergic agonist isoproterenol (Iso) on cells of the inner stripe portion of the rabbit outer medullary collecting duct (OMCDi) grown in primary culture were examined using whole cell patch-clamp techniques and measurements of intracellular pH (pHi) and Ca2+. Iso (10(-6) M) increased the cellular Cl- conductance, and this effect was mimicked by treatment of the cells with dibutyryladenosine 3',5'-cyclic monophosphate (cAMP, 10(-5) M) or protein kinase A (PKA, 0.4 U/ml). Iso did not alter the baseline pHi, but it did increase the activity of both the Cl-/HCO3- antiporter and the H(+)-adenosinetriphosphatase (H(+)-ATPase). The increase in Cl-/HCO3- antiporter rate was mimicked by dibutyryl-cAMP plus 3-isobutyl-1-methylxanthine (cAMP + IBMX, 10(-4) M + 10(-5) M). However, the Iso-induced stimulation of the H(+)-ATPase activity was not mimicked by cAMP + IBMX. Measurements of intracellular Ca2+ showed that Iso also increased intracellular Ca2+ levels. This response was not dependent on extracellular Ca2+, nor did cAMP + IBMX appreciably alter intracellular Ca2+. Consequently, we postulate that beta-adrenergic agonists are potential stimulators of OMCDi H+ secretion. These agonists stimulate cellular HCO3- efflux through a signal transduction pathway involving cAMP and PKA. However, a different signal transduction pathway appears to mediate the stimulation of cellular H+ efflux. This second pathway may involve an elevation of intracellular Ca2+.


Subject(s)
Hydrogen/metabolism , Kidney Tubules, Collecting/metabolism , Receptors, Adrenergic, beta/physiology , 1-Methyl-3-isobutylxanthine/pharmacology , Animals , Bucladesine/pharmacology , Calcium/metabolism , Cells, Cultured , Chlorides/physiology , Electrophysiology , H(+)-K(+)-Exchanging ATPase/physiology , Hydrogen-Ion Concentration , Intracellular Membranes/metabolism , Isoproterenol/pharmacology , Kidney Medulla , Kidney Tubules, Collecting/cytology , Kidney Tubules, Collecting/physiology , Protein Kinases/pharmacology , Rabbits
12.
Am J Physiol ; 263(6 Pt 2): F1004-10, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1336307

ABSTRACT

Whole cell patch-clamp techniques were used to characterize the electrophysiological properties of cells from the inner stripe portion of the rabbit outer medullary collecting duct (OMCDi) grown in primary culture. With pipette and bathing solutions mimicking intracellular and extracellular fluid, the resting membrane voltage was -30 to -40 mV. The whole cell conductance exhibited slight outward rectification, and at the resting membrane voltage the cell conductance averaged 2.58 +/- 0.49 nS (n = 17). The major conductive ion species was Cl-. The Cl- conductance was also found to have a significant permeability to HCO3- and was inhibited by the Cl(-)-channel blockers diphenylamine carboxylic acid and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. A small K+ conductance was also present, but no Na+ conductance was detected. Current generated by the H(+)-adenosinetriphosphatase (H(+)-ATPase) was quantitated. This current was dependent on the presence of ATP in the pipette. Dicyclohexylcarbodiimide, N-ethylmaleimide, and bafilomycin A1, inhibitors of the vacuolar H(+)-ATPase, also reduced this outward current in an ATP-dependent manner. The inhibitor-sensitive component of the outward current, a measure of the current generated by the H(+)-ATPase, was in the range of 35-100 pA/cell.


Subject(s)
Kidney Tubules, Collecting/physiology , Animals , Cells, Cultured , Chlorides/physiology , Electrophysiology , H(+)-K(+)-Exchanging ATPase/physiology , Kidney Medulla , Kidney Tubules, Collecting/cytology , Rabbits
14.
Breast Cancer Res Treat ; 16(3): 273-8, 1990 Oct.
Article in English | MEDLINE | ID: mdl-2085677

ABSTRACT

We have previously described an in vitro immunohistochemical test employing anti-receptor antibodies, for demonstrating the nuclear binding characteristics of estrogen receptors (ER) in breast carcinomas. Based on a retrospective analysis of twenty-five patients with estrogen receptor-positive (ER+) breast cancer who were treated with hormone therapy and whose clinical responses were evaluable, we were able to demonstrate that this test may be valuable to predict which, among the ER+ tumors (whether or not they are progesterone receptor positive, PR+), are likely to respond to hormone therapy and which may fail. While tumors in which ER exhibited abnormalities in nuclear binding behavior (ligand-independent nuclear binding or no nuclear binding) failed hormone therapy (16 out of 19 patients), those in which nuclear binding of ER appeared normal (ligand-dependent) in the in vitro test, responded to hormone therapy (5/6 patients). While our previous report dealt with the procedural details, specificity of the reagents, and the design of the study, this report addresses the clinical aspects of this study and response correlation.


Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/metabolism , Cell Nucleus/metabolism , Neoplasm Proteins/metabolism , Receptors, Estrogen/metabolism , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Drug Resistance , Female , Humans , Male , Neoplasm Metastasis , Neoplasms, Hormone-Dependent/metabolism , Retrospective Studies , Tamoxifen/therapeutic use
15.
J Natl Cancer Inst ; 80(10): 756-61, 1988 Jul 20.
Article in English | MEDLINE | ID: mdl-3290497

ABSTRACT

Neither estrogen dependence nor clinical response to hormone therapy of the tumor is guaranteed by the mere presence of receptors for estrogens (ERs) or progesterones (PRs). Immunohistochemical staining of ER with polyclonal anti-ER antibodies after in vitro transformation has enabled the identification of two types of defective ER among human breast cancers: those that are unable to bind to the nucleus in a hormone-filled state and those that bind to the nucleus as naked ER. Preliminary clinical correlation studies demonstrate that a subclassification of ER(+) tumors based on functional abnormalities of ER may predict refractoriness to hormone therapy.


Subject(s)
Breast Neoplasms/metabolism , Cell Nucleus/metabolism , Neoplasms, Hormone-Dependent/metabolism , Receptors, Estrogen/metabolism , Antibodies, Monoclonal , Breast Neoplasms/therapy , Breast Neoplasms/ultrastructure , Fluorescent Antibody Technique , Humans , Molybdenum , Neoplasms, Hormone-Dependent/therapy , Quality Control
16.
Steroids ; 51(5-6): 425-39, 1988.
Article in English | MEDLINE | ID: mdl-3242169

ABSTRACT

The 'two-step' model proposed by Jensen and his collaborators for explaining estrogen action conceptualized hormone-free estrogen receptors (ER) to be cytoplasmic, and hormone-filled, transformed ER to be nuclear. Applying monoclonal antibodies which recognized epitopes in ER and formaldehyde-fixed tissues, King et al demonstrated exclusively nuclear staining in target tissues utilizing immunoperoxidase technique. Recently these antibodies have become commercially available enabling other investigators to conduct studies. In this report, using these monoclonal antibodies we have demonstrated that a change in the concentration of formaldehyde alters the staining pattern yielding cytoplasmic instead of nuclear staining in calf uterus, MCF-7 cells, and ER(+) human breast cancer. In addition, neutralization of the antibody activity was not achieved with freshly prepared ER(+) cytosols. Formaldehyde-treated cytosols were essential. These results ought to caution investigators in determining in vivo location of antigens based on the staining pattern obtained in fixed tissues. Furthermore, this effect of formaldehyde on estrogen receptors may be applicable to other steroid hormone receptors.


Subject(s)
Antibodies, Monoclonal , Cell Nucleus/analysis , Fixatives , Receptors, Estrogen/analysis , Animals , Cattle , Formaldehyde , Humans , Immunohistochemistry , Reproducibility of Results
17.
J Nurs Adm ; 18(5): 31-3, 1988 May.
Article in English | MEDLINE | ID: mdl-3367228

ABSTRACT

In 1983, the newly implemented prospective payment system reduced the length of hospital stays, thereby curtailing many of the services that patients traditionally received. The Division of Nursing at Memorial Medical Center of Long Beach, California identified many patient needs that were no longer being addressed during the inhospital stay and began looking for a way for nurses to meet these needs on an outpatient basis. The result was the formation of Memorial Medical Center's Community Nursing Center.


Subject(s)
Community Health Nursing/organization & administration , Nursing Staff, Hospital , Outpatient Clinics, Hospital/organization & administration , California , Hospital Bed Capacity, 500 and over , Humans , Patient Education as Topic
18.
Eur J Cancer Clin Oncol ; 20(10): 1261-77, 1984 Oct.
Article in English | MEDLINE | ID: mdl-6386482

ABSTRACT

An immunohistochemical (IF) method utilizing polyclonal antibodies to cytoplasmic estrogen receptors (ER) and a tissue processing procedure which allows in situ precipitation of cytoplasmic ER without denaturation of antigenic sites were first described in this journal. In this report we present data on the immunohistochemical classification of 153 cases of human breast carcinomas (123 frozen sections, 28 cytological specimens) as ER-positive (IF+) or ER-negative (IF-). Results on correlation between the biochemical vs immunohistochemical assessment of the ER status and the relationship between quantity of ER and the proportion of IF+/IF- tumor cells are presented. In addition, a procedure which has been successfully applied to study in vitro translocation of cytoplasmic ER in cryostat sections of human breast carcinomas is described. A positive correlation between ER translocatability and status of progesterone receptors (PR) was obvious. All biochemical and immunohistochemical data are collectively reviewed and the question of whether the antibodies are detecting Type I ER is examined.


Subject(s)
Breast Neoplasms/analysis , Receptors, Estrogen/analysis , Antibodies/immunology , Breast Neoplasms/pathology , Cell Nucleus/analysis , Cytoplasm/analysis , Female , Fluorescent Antibody Technique , Humans , Menopause , Receptors, Estrogen/immunology , Receptors, Progesterone/analysis
19.
Am J Hosp Pharm ; 41(7): 1352-8, 1984 Jul.
Article in English | MEDLINE | ID: mdl-6465150

ABSTRACT

The effect of an automated bedside dispensing machine on medication errors was studied on a 32-bed surgical unit of an 848-bed hospital. The experimental system (McLaughlin Dispensing System) included at each patient's bedside a locked medication cabinet that was electronically programmed to allow the nurse access to doses due at a particular time. The control system was a decentralized unit dose system. A crossover study design with random assignment of subjects and treatments was used. In the 14-day study period, nurses were observed by a pharmacist for 28 five-hour periods as they administered medications on the day and evening shifts. The mean error rates were significantly different--10.6% for the experimental system and 15.9% for the control system. Wrong time errors were the most common type. No significant differences were found between day and evening shifts or workloads of individual nurses. There was no treatment order effect. The error rate was significantly lower for the automated dispensing system than for the system using unit doses dispensed from a satellite pharmacy. Automated dispensing systems may be useful in reducing errors in administration time and dose omissions.


Subject(s)
Medication Errors , Medication Systems, Hospital , Pharmaceutical Preparations/administration & dosage , Computers , Hospital Bed Capacity, 500 and over , Humans , Pharmacy Service, Hospital
20.
Hum Pathol ; 11(6): 675-7, 1980 Nov.
Article in English | MEDLINE | ID: mdl-7450740

ABSTRACT

A 57 year old man presented with a two week history of right sided chest pain. An admission chest x-ray examination revealed a 2 by 1 cm. lytic lesion of the ninth right rib and right sided pleural effusion. The patient underwent a right thoracotomy during which 700 cc. of blood tinged pleural fluid was drained and a 10 cm. segment of the right ninth rib was excised. The rib lesion showed classic features of eosinophilic granuloma of bone. Cytologic examination of the pleural fluid disclosed abundant histiocytes and eosinophils, consistent with eosinophilic granuloma invading the pleural space. This case is unusual because of the age of the patient and the development of a pleural effusion secondary to the rupture of the tumor into the pleural cavity, the latter not previously reported in the literature.


Subject(s)
Eosinophilic Granuloma/complications , Pleural Effusion/etiology , Ribs/pathology , Age Factors , Eosinophilic Granuloma/pathology , Humans , Male , Middle Aged
SELECTION OF CITATIONS
SEARCH DETAIL
...