Biomedical subjects

C Ince

Publications and source records attributed to C Ince.

At least 37 records · Page 2Linked to original sources

PubMed1992

Optical spectroscopic imaging for non-invasive evaluation of tissue oxygenation.

Altered rates of oxygen delivery and uptake between and possibly within different organs occur during critical illness. The mechanisms governing this heterogeneity are as yet not fully understood and techniques directed at being able to map the course of oxygen to the mitochondria to produce ATP, the main molecule needed to drive energy requiring processes in the cell, would give valuable information about the mechanisms underlying organ dysfunction during disease. Oxidative phosphorylation occurring in the mitochondria is the main site for the production of ATP in mammalian cells. Metabolic substrates, ADP and Pi, and O2 are the ingredients needed to produce ATP. Due to the central role of oxidative phosphorylation in the metabolism of the cell, much attention has been directed at developing non-invasive techniques to measure intermediates of the oxidative phosphorylation in tissue as an indication of the metabolic state of tissue. One such method enables mapping the distribution of tissue hypoxia by use of a fluorescence technique based on the measurements reduced nicotine amide dinucleotide (NADH). NADH is situated at the high-energy side of the respiratory chain and during tissue hypoxia accumulates in concentration because less NADH is oxidized to NAD+. Excitation of NADH by 366 nm light produces, unlike NAD+, fluorescence at 460 nm light. Previously, however, producing images of NADH fluorescence distribution in tissue has been limited to saline perfused in vitro models. We recently undertook to develop an NADH videoflurometer sensitive enough to NADH fluorescence in vivo (Ince C, Bruining HA (1991) Optical Spectroscopy for the measurement of tissue hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)

Fluorometry↗

PubMed1991

A phase I study of a combination of allopurinol, 5-fluorouracil and leucovorin followed by hydroxyurea in patients with advanced gastrointestinal and breast cancer.

Twenty patients with advanced carcinomas of the colorectum, pancreas, stomach, and breast were enrolled in a Phase I study of a sequential administration of 5-fluorouracil-leucovorin (FU-LV) combination followed by hydroxyurea (HU) with allopurinol protection (HALF regimen). As a weekly regimen for 6 weeks, followed by a rest period of 2 weeks, FU was administered intravenously (i.v.) during infusion of a 2-hour i.v. infusion of LV at a dose of 500 mg/m2. Six hours following the FU-LV combination, HU (1 gm/m2) was administered orally. Allopurinol (300 mg every 8 hours, orally) was given the day before and on the day of the administration of the FU-LV combination. The starting dose of FU was 300 mg/m2, with escalations to 900 mg/m2. Mucositis, diarrhea, and hematologic toxicities were mild and sporadic with FU doses up to 750 mg/m2 and occurred in patients who had received prior treatment with FU and/or radiotherapy. Dose-limiting neurocerebellar toxicity was observed in 2 out of 6 patients who received a FU dose of 900 mg/m2. Three additional patients experienced moderate neuromotor toxicity at this dose level. Among 17 patients evaluable for response, partial responses were seen in 3 of the 9 patients with colorectal cancer, 1 of the 3 patients each with carcinoma of breast and pancreas. Three of the 5 responses occurred in patients who had received prior treatment with FU and/or radiation therapy. An FU dose of 750 mg/m2 is recommended for a Phase II trial of the HALF regimen.

Aged↗

PubMed1990

A micro-perfusion chamber for single-cell fluorescence measurements.

A versatile closed micro-perfusion chamber designed for single-cell fluorescence measurements under maximum microscopic magnification is described. Glass coverslips with adherent cells can be attached to the top or bottom of the chamber, depending on whether an inverted or an upright microscope is used. Eight conical holes drilled in the side of the chamber serve for the insertion of plugs with attachments for perfusion, rapid injection of small amounts of reagents, temperature measurements or for heating the interior of the chamber. Materials used in the construction of the chamber are non-toxic and resistant to standard sterilization procedures. Perfusion and temperature properties of the chamber are described. Single cell fluorescence measurements are presented in human monocyte-derived macrophages in which NAD(P)H and intracellular calcium are measured.

Calcium↗

PubMed1990

Selective decontamination to reduce gram-negative colonisation and infections after oesophageal resection.

181 patients undergoing resection of the oesophagus for carcinoma were randomised to receive selective decontamination (test group) or conventional perioperative antibiotic prophylaxis (controls). 114 patients were finally included in the study: 12 of 56 test patients had 18 infections, whereas 32 of 58 controls acquired 51 infections. Colonisation with aerobic gram-negative microorganisms, and the number of postoperative respiratory tract infections were significantly lower in the test patients. The postoperative therapeutic use of antibiotics was significantly lower in the test group. No endogenous infections were caused by gram-negative bacilli in the test group. Selective decontamination reduces colonisation with gram-negative bacilli and postoperative infections after resection of the oesophagus.

Adult↗

PubMed1990

Heterogeneous NADH fluorescence during post-anoxic reactive hyperemia in saline perfused rat heart.

In the present study epicardial NADH fluorescence photographs were taken of rat hearts during dynamic transitions of oxygen content of the myocardium. Hearts were perfused in a Langendorff set-up where it was possible to switch between low and high-pO2 perfusates. NADH fluorescence photographs were taken with a suitable fluorescence set-up and photo negatives digitized and analyzed by use of a computer. Restoration of perfusion with a high-pO2 solution resulted in a reactive hyperemic flow being established. Prior to the occlusion being lifted high NADH fluorescence was observed. Reactive hyperemic flow was associated with heterogenic NADH fluorescence patterns which diminished as control flow was restored. The patterns observed during reactive hyperemia were identical to those observed when tissue oxygen was restored by high-pO2 perfusion following high flow hypoxia achieved by low-pO2 perfusion. This study shows that heterogenic epicardial flow patterns are associated with reactive hyperemia.

Animals↗

PubMed1990

Effects of selective decontamination on gram-negative colonisation, infections and development of bacterial resistance in esophageal resection.

Patients undergoing an esophageal resection because of carcinoma are at risk of developing postoperative respiratory tract infections. These patients were studied with respect to preceding colonisation with gram-negative bacilli and the effect of selective decontamination (SD) in decreasing this phenomenon, thereby reducing gram-negative infections. We randomised prospectively 114 patients into a test group receiving SD-medication (n = 56) and a control group receiving conventional prophylaxis. Postoperatively, all patients were admitted to the intensive care unit and mechanically ventilated. The preoperative administration of SD-medication resulted in adequate decontamination within 3-4 days in most patients, and SD could prevent gram-negative colonisation and infections effectively. Discontinuation of SD showed gram-negative (re-)colonisation, and resulted in 12 infections in 4 patients having late complications. This indicates that prolonged use in these patients might be beneficial. This new antibiotic prophylaxis proved effective, without causing an increase in bacterial resistance.

Anti-Bacterial Agents↗

PubMed1990

Design and validation of an indicator gas injector for multiple gas washout tests in mechanically ventilated patients.

A device to produce a stepwise indicator gas fraction variation to initiate a washout test in mechanically ventilated patients is described. The device, which can be used in conjunction with the commonly used Siemens-Elema series 900 ventilators, is based on simple, off-the-shelf technology. It features the simultaneous use of two indicator gases (so that the influence of diffusion processes in the gas exchange to the patient can be measured) and maintains a nearly constant FIO2 during a washout procedure. With this indicator gas injector, the transition time of the indicator gas fraction at the beginning of a washout proved to be short enough to detect ventilation inhomogeneity by visual inspection of the washout curves. Functional residual capacity measurements using this device are presented on a test lung with known volume, on healthy volunteers, and on critically ill patients.

Adolescent↗

PubMed1989

Early events in the antiproliferative action of tumor necrosis factor are similar to the early events in epidermal growth factor growth stimulation.

The process of TNF-induced cytotoxicity is complex but appears to be mediated through a TNF-specific cell surface receptor. Recent evidence suggests that TNF action on tumor cells may be antagonized by epidermal growth factor (EGF) and other EGF-receptor modulatory peptides implicating a role for EGF-R in the process of TNF-induced cytotoxicity. In the present report, we investigated the biochemical actions of TNF on several biochemical events known to occur in the process of EGF signal transduction in intact cells. The actions of TNF were compared directly to those of EGF in both TNF-sensitive and -resistant tumor cell lines. In TNF-sensitive ME-180 cervical carcinoma cells, TNF (20 ng/ml) stimulated the tyrosine protein kinase activity of the EGF-receptor (EGF-R) fivefold when measured by receptor autophosphorylation in an immune complex kinase assay. TNF activation of EGF-R kinase activity in ME-180 was measurable 10 min after TNF incubation and enzymatic activity remained elevated 20 min after TNF addition. Activation of the receptor by TNF correlated with increased 32P incorporation into EGF-R protein when receptor was immunoprecipitated from 32P-equilibrated cells following a 20 min incubation with TNF. Acid hydrolysis of EGF-R protein isolated from TNF-treated ME-180 cells demonstrates an increase in the phosphotyrosine content of EGF-R when compared to receptor isolated from untreated cells. The results suggest that TNF increased EGF-R tyrosine protein kinase activity and the state of EGF-receptor tyrosine phosphorylation in a manner similar to that reported for EGF. However, TNF does not appear to be structurally related to EGF since TNF was unable to directly activate EGF-R when incubated with extensively washed immunoprecipitates of EGF-R. In TNF-resistant T24 bladder carcinoma cells, TNF failed to alter EGF-R tyrosine protein kinase activity although both EGF and phorbol ester were shown to modulate the enzymatic activity of the receptor in these cells. These results indicate that the ability of TNF to modulate EGF-R kinase in target cells may correlate with its cytotoxic actions on TNF-sensitive tumor cells. Other biochemical activities associated with the induction or regulation of cellular growth were examined in TNF- or EGF-treated tumor cells. EGF stimulated a rapid 8-16-fold increase in the expression of the proto-oncogene c-myc when analyzed by dot-blot analysis of total cellular RNA or Northern blot hybridization of polyadenylated RNA. TNF treatment failed to alter c-myc expression in ME-180 cells when analyzed by either technique.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell Division↗

PubMed1988

Reduction to homozygosity at the SIS/PDGF-2 locus in human mesenchymal tumors.

Enhanced expression of the human SIS/PDGF-2 gene has been reported in a number of human cell lines, sarcomas, and glioblastomas. We have analyzed the SIS/PDGF-2 gene for structural alterations in fresh human tumors. DNA samples from 79 patients with solid tumors (63 mesenchymal tumors, 12 lung carcinomas, 4 breast carcinomas) were examined and compared with DNA samples from 50 leukemia patients and 14 unrelated individuals without malignant neoplasms. When DNA samples were digested with a HindIII restriction endonuclease, Southern blot analysis demonstrated two distinct bands (21kb and 18kb) after hybridization to the SIS/PDGF-2 gene probe. A pedigree analysis of a 43-member family indicated that these allelic variants segregated in a Mendelian fashion. There was, however, tumor specific allele loss in 18% of the mesenchymal tumors analyzed, which may indicate a common etiology for this tumor type.

Cell Line↗

PubMed1988

Brain infarction in sickle cell anemia: magnetic resonance imaging correlates.

Brain infarction is a well-known but poorly understood complication of sickle cell disease. Seventy-three sickle cell disease patients underwent neurological examinations and high-field, heavily T2-weighted axial cranial magnetic resonance image (MRI) scanning. Eighteen of the 73 had a history of stroke, defined as an acute, focal neurological sign lasting longer than 1 hour; in the event of a convulsive onset, an MRI abnormality as a correlate was necessary. Sixteen of the 18 stroke patients demonstrated focal MRI abnormalities consistent with arterial borderzone infarctions. Fifty-five of the 73 patients had no history of stroke. Six of the 55 (11%) had focal MRI abnormalities suggesting previous subclinical stroke. Five of these lesions were in borderzone regions. The distinguishing feature in 21 of the 22 patients with MRI abnormalities was the predilection for lesions in the high cortical convexity, in the general regions of arterial borderzones between the major cerebral arteries and adjacent deep white matter. The pattern of MRI lesions suggests two pathogenetic mechanisms: (1) proximal large-vessel disease with inadequate cerebral perfusion (distal field insufficiency syndrome) and (2) distal small-vessel disease (sludging syndrome).

Adolescent↗

PubMed1988

Extracellular ATP induces a large nonselective conductance in macrophage plasma membranes.

Extracellular ATP in its tetra-anionic form (ATP4-) induces ion fluxes and membrane depolarization in the mouse macrophage-like cell line J774.2 and in resident mouse macrophages. We analyzed the effects of extracellular ATP4- by both patch-clamp and intracellular microelectrode techniques. Whole-cell patch-configuration membrane potential measurements on J774.2 cells revealed that ATP4- -induced depolarization occurred within 40 ms of pulsed application of ATP and was completely reversible. The depolarizations were accompanied by a dramatic increase in membrane conductance and showed no sign of adaptation to ATP over a period of 30 min. At 5 mM total ATP (ATPt) the whole-cell conductance was approximately 10 nS, and an upper limit of 20 pS for a single-channel conductance has been established. The reversal potential associated with the ATP-induced depolarization at asymmetric K+, Na+, Ca2+, and Cl- concentrations across the membrane was 0 mV. In patch-clamped cells depolarization was complete at 20 microM ATP4-, and repolarization from full depolarization occurred in approximately 5 s. In contrast, in intact cells measured by microelectrode impalement, complete depolarization occurred at approximately 2 mM ATP4- and repolarization was much slower (approximately 100 min). These findings indicate that the changes in intracellular ionic composition that occur after ATP treatment affect the rate of cell repolarization. At lower concentrations of ATP, potassium conductances modulated the depolarizing effect of ATP. ATP also depolarized mouse peritoneal macrophages, but a variant cell line (ATPR B2), derived from J774.2 cells by prolonged exposure to ATP, was insensitive to ATP. Our results provide a membrane electrophysiological description and analysis of a large nonselective plasma membrane conductance of macrophages induced by extracellular ATP.

Adenosine Triphosphate↗

PubMed1988

Phagocytosis by human macrophages is accompanied by changes in ionic channel currents.

The present study has shown that changes in ionic channel currents accompany the phagocytosis of particles by mononuclear phagocytes. The patch-clamp technique in the cell-attached configuration was applied to human monocyte-derived macrophages to measure the activity of single transmembrane ionic channels in intact cells. During such measurements, IgG-opsonized and non-opsonized latex particles were offered for phagocytosis under continuous video-microscopical observation. Single particles were presented to the phagocytes at a membrane location some distance from that of the patch electrode. After a lag period following particle attachment, enhanced inward and outward time-variant single channel currents coinciding with particle engulfment were observed. On the basis of current-voltage characteristics and membrane potential measurements, the outward-directed channels were identified as K+ channels. Phagocytosis was also accompanied by slow transient changes in background membrane currents, probably due to changes in the membrane potential of the phagocytosing cell. Phagocytosis of IgG-coated latex particles differed from phagocytosis of uncoated or albumin-coated particles by a shorter lag time between particle attachment and the onset of enhanced ionic channel activity.

Electric Conductivity↗

PubMed1988

Bcl-1 gene rearrangements in B cell lymphoma.

We analyzed 50 B cell lymphoma samples by Southern blot analysis, using the bcl-1 and heavy chain immunoglobulin (JH) probes with two or more restriction endonucleases. All samples showed JH rearrangement, and three samples (two diffuse small lymphocytic lymphomas and one diffuse large cell lymphoma probably transformed from a diffuse small lymphocytic lymphoma) demonstrated rearranged bcl-1 sequences. The three samples showed the t(11;14)(q13;q32) chromosome translocation, and all three contained rearranged JH fragments that comigrated with the rearranged bcl-1 fragment. The breakpoint of the translocation occurred within a 1.6-kb region on chromosome 11 in the three cases. Two of the three patients had primary refractory disease. Two of the three patients had gastrointestinal involvement. Bcl-1 rearrangement may identify an unusual subset of patients with primary refractory disease with gastrointestinal involvement. It may also describe a unique subset of large cell lymphoma patients transformed from diffuse small cell histology.

B-Lymphocytes↗

PubMed1987

Intracellular K+, Na+ and Cl- concentrations and membrane potential in human monocytes.

The relationship between the resting membrane potential and the intracellular ionic concentrations in human monocytes was investigated. Cell volume, cell water content, and amount of intracellular K+, Na+, and Cl- were measured to determine the intracellular concentrations of K+ (Ki), Na+ (Nai) and Cl- (Cli) of monocytes, and of lymphocytes and neutrophils. Values found for monocytes were similar to those for neutrophils, i.e., cell volumes were 346 and 345 micron3, respectively, cell water content 78%, and Ki, 128 and 125, Nai, 24 and 26, and Cli, 102 and 103 mmol/l cell water, respectively. Lymphocytes, however, had different values: 181 micron3 cell volume, 77% cell water content, and for Ki, Nai, and Cli, 165, 37, and 91 mmol/l cell water, respectively. The resting membrane potential of cultured human monocytes (range -30 to -40 mV), determined by measurement of the peak potential occurring within the first milliseconds after microelectrode entry, was most dependent on extracellular K+, followed by Cl-, and Na+. The membrane permeability ratio of Cl- to K+ was estimated by use of the constant field equation to be 0.23 (range 0.22 to 0.30).

Body Fluids↗

PubMed1987

Ionic channels and membrane hyperpolarization in human macrophages.

Microelectrode impalement of human macrophages evokes a transient hyperpolarizing response (HR) of the membrane potential. This HR was found to be dependent on the extracellular concentration of K+ but not on that of Na+ or Cl-. It was not influenced by low temperature (12 degrees C) or by 0.2 mM ouabain, but was blocked by 0.2 mM quinine or 0.2 mM Mg2+-EGTA. These findings indicate that the HR in human macrophages is caused by the activation of a K+ (Ca2+) conductance. Two types of ionic channels were identified in intact cells by use of the patch-clamp technique in the cell-attached-patch configuration, low and high-conductance voltage-dependent K+ channels. The low-conductance channels had a mean conductance of 38 pS with Na+-saline and 32 pS with K+-saline in the pipette. The high-conductance channels had a conductance of 101 and 114 pS with Na+- and K+-saline in the pipette, respectively. Cell-attached patch measurements made during evocation of an HR by microelectrode penetration showed enhanced channel activity associated with the development of the HR. These channels were also high-conductance channels (171 pS with Na+- and 165 pS K+-saline in the pipette) and were voltage dependent. They were, however, active at less positive potentials than the high-conductance K+ channels seen prior to the microelectrode-evoked HR. It is concluded that the high-conductance voltage-dependent ionic channels active during the HR in human macrophages contribute to the development of the HR.

Cell Membrane↗

PubMed1987

Prenatal diagnosis of sickle hemoglobinopathies: the experience of the Columbia University Comprehensive Center for Sickle Cell Disease.

We report here an evaluation of 55 pregnancies at risk for a sickle hemoglobinopathy prenatally diagnosed by restriction-endonuclease analysis, with the endonucleases MstII and HpaI, of amniocyte DNA. The diagnosis was completed in all cases. Eleven fetuses were predicted to be affected, of which six were terminated. Forty-one of the 55 cases were confirmed. One false-negative was reported in a case predicted to be hemoglobin AS but that was determined to be hemoglobin SS at birth. We estimate that the 55 cases represent only 5% of the pregnancies at risk for a sickle hemoglobinopathy in the New York metropolitan area during the study period. We conclude that the prenatal diagnosis of sickle hemoglobinopathies by molecular methods is reliable. However, the efficiency of utilization and effectiveness of prenatal testing is dependent on the early prospective identification of couples at risk and on the education of communities concerning the significant morbidity of the sickle hemoglobinopathies and the reproductive choices now available to them.

Anemia, Sickle Cell↗

PubMed1986

Intracellular microelectrode measurements in small cells evaluated with the patch clamp technique.

Microelectrode penetration of small cells leads to a sustained depolarization of the resting membrane potential due to a transmembrane shunt resistance (Rs) introduced by the microelectrode. This has led to underestimation of the resting membrane potential of various cell types. However, measurement of the fast potential transient occurring within the first few milliseconds after microelectrode penetration can provide information about pre-impalement membrane electrophysiological properties. We have analyzed an equivalent circuit of a microelectrode measurement to establish the conditions under which the peak of the impalement transients (Ep) approaches the pre-impalement resting membrane potential (Em) of small cells most closely. The simulation studies showed that this is the case when the capacitance of the microelectrode is low and the membrane capacitance of the cell high. In experiments performed to assess the reliability of Ep as a measure of Em, whole-cell patch clamp measurements were performed in the current clamp mode to monitor, free from the effects of Rs, Em in cultured human monocytes. Microelectrode impalement of such patch clamped cells and measurement of Ep made it possible to detect correlation between Ep and Em and showed that for small cells such as human monocytes Ep is on average 6 mV less negative than the resting membrane potential.

Cell Membrane↗

PubMed1985

A teflon culture dish for high-magnification microscopy and measurements in single cells.

A simple tissue culture dish is described which permits the use of oil immersion optics (X 100) while maintaining access to cells in culture. In combination with a micro-CO2-incubator (Ince et al. 1983) long-term as well as short-term experiments can be performed under microscopical control. The teflon dish is re-usable, resistant to sterilization procedures, and easy to assemble. Standard glass coverslips with adherent cells are secured to the dish by an aluminium ring with a bayonet fitting. Both phase-contrast and interference-contrast microscopy with high magnification can be used. The dish is of particular use in electrophysiological investigations where the patch-clamp technique is used and frequent exchange of cultures is required. With this unit, single ion channel activity in cultured human monocytes was measured. The dish has also been used in studies in which membrane potential measurements were performed with glass microelectrodes in small cells under culture conditions. The applications described include video time-lapse sequences of phagocytosis with micro-organisms presented to mouse peritoneal macrophages with broken-tipped pipettes. This easy to use, multi-purpose dish offers the cell physiologist a valuable aid for the manipulation and observation of single cells in culture.

Animals↗