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PubMed1994

2-Fluoroadenosine uptake by erythrocytes and endothelial cells studied by 19F-NMR.

Transport and phosphorylation of 2-fluoroadenosine (F-AR) were studied in human erythrocytes and porcine aortic endothelial cells by 19F-nuclear magnetic resonance (NMR) spectroscopy. F-AR (590 microM) added to a human erythrocyte suspension (15% hematocrit) was rapidly incorporated into adenine nucleotides at a rate of 38 nmol.min-1.ml red blood cells-1. Intracellular F-AR could be distinguished from extracellular F-AR due to a chemical shift difference of 0.43 +/- 0.03 ppm (n = 5 experiments). Compared with F-AR, fluoro-ATP purified by high-performance liquid chromatography (HPLC) exhibited a chemical shift of -0.052 ppm, which was too small to differentiate intracellular F-AR and fluoro-ATP in vivo. F-AR uptake was decreased by inhibition of membrane transport with dipyridamole (25 microM) or blockade of adenosine kinase by iodotubercidin (10 microM). The time course of F-AR uptake suggested that the rate-limiting step was not membrane transport but the intracellular phosphorylation by adenosine kinase. In porcine aortic endothelial cells grown on microcarrier beads and perfused within the magnet, there was a linear relation between the F-AR concentration applied (2, 4, 8, or 32 microM) and net uptake measured (27-827 pmol.min-1.mg-1). Intra- and extracellular fluoroadenine compounds were separated by 0.12 ppm, and HPLC analysis confirmed F-AR conversion to fluoroadenine nucleotides. Our findings demonstrate that cellular transport and metabolism of F-AR can be noninvasively studied and analyzed by 19F-NMR.

Adenosine↗
PubMed1993

Islet cell cytoplasmic autoantibody reactivity to glutamate decarboxylase in insulin-dependent diabetes.

Individuals with or at risk for insulin-dependent diabetes (IDD) frequently have autoantibodies against an islet cell cytoplasmic (ICA) antigen thought to be a sialoglycolipid. However, we now report that preabsorption of ICA-positive sera with recombinant glutamate decarboxylase (human GAD 65 and/or GAD 67) reduced or blocked the ICA reactivity of 5/18 (27%) new-onset IDD patients and 7/18 (39%) prediabetics. Interestingly, nondiabetic subjects with ICA of > or = 5 yr in duration had GAD-reactive ICA significantly more often (16/24, 67%, P < 0.04) than the diabetic groups. ICA reactivity to GAD was not related to serum ICA titer nor the age of the individual, and in all cases tested was blocked by GAD 65 or GAD 67 with equivalent efficiency. The ICA observed in 21/25 (84%) IDD patients with ICA long after clinical onset of disease (9-42 yr) was reactive to GAD. A natural history analysis of three individuals showed conversions from ICA which was reactive to GAD to a non-GAD-reactive ICA nearer to their clinical onsets of IDD. This study further defines the autoantigens reactive to ICA, and suggests that, whereas ICA that are not reactive to GAD may identify an advanced and more prognostic lesion, GAD-reactive ICA may typify the early or inductive lesion that may or may not progress to clinically significant beta cell injury.

Adolescent↗
PubMed1995

The origin of the myofibroblasts in breast cancer. Recapitulation of tumor environment in culture unravels diversity and implicates converted fibroblasts and recruited smooth muscle cells.

The origin of myofibroblasts in stromal reaction has been a subject of controversy. To address this question definitively, we developed techniques for purification and characterization of major stromal cell types. We defined a panel of markers that could, in combination, unequivocally distinguish these cell types by immunocytochemistry, iso-electric focusing, immunoblotting, and two-dimensional gel electrophoresis. We then devised an assay to recapitulate in culture, within two weeks of incubation, critical aspects of the microenvironment in vivo including the typical tissue histology and stromal reaction. When confronted with tumor cells in this assay, fibroblasts readily converted into a graded pattern of myogenic differentiation, strongest in the immediate vicinity of tumor cells. Vascular smooth muscle cells (VSMC), in contrast, did not change appreciably and remained coordinately smooth muscle differentiated. Midcapillary pericytes showed only a slight propensity for myogenic differentiation. Analysis of ten primary tumors implicated converted fibroblasts (10/10), vascular smooth muscle cells (4/10), and pericytes (1/10) in the stromal reaction. Tumor cells were shown to specifically denude the venules both in culture and in vivo, explaining the VSMC phenotype in the stroma. The establishment of this assay and clarification of the origin of these cells pave the way for further analysis of the mechanisms of conversion, and of the consequence of such heterogeneity for diagnosis and treatment.

Biomarkers↗
PubMed1975

Renal sodium-potassium adenosine triphosphatase. Optical localization and x-ray microanalysis.

The distribution of sodium-potassium adenosine triposphatase (Na-K-ATPase) activity in kidney sections has been studied by a method based on the hydrolysis of p-nitrophenyl phosphate in alkaline medium containing dimethyl sulfoxide. The products at each stage in the reaction sequence have been subjected to electron probe microanalysis. The initial product was identified as a mixture of KMgPO4 and Mg(PO4)2, and sequential analysis demonstrated the linearity of conversion of this product to a visible form. In human, rabbit and rat kidneys the distribution of activity was found to be essentially identical, with highest levels located in thick ascending limbs and distal convoluted tubules. The initial reaction was completely potassium dependent and was inhibited by ouabain in concentrations reflecting the relative sensitivity of microsomal Na-K-ATPase in each species. Measurement of initial product phosphorus by means of the electron probe is presented as a practical technique for direct quantitation of Na-K-ATPase activity in identified tubule segments.

Adenosine Triphosphatases↗
PubMed1997

Calcium absorptive effects of vitamin D and its major metabolites.

The absorptive response to graded doses of vitamin D3, 25(OH)D, and 1,25(OH)2D was measured in healthy adult men after treatment periods of eight, four, and two weeks, respectively. While no relationship was found between baseline absorption and serum vitamin D metabolite levels, all three vitamin D compounds significantly elevated 45Ca absorption from a 300 mg calcium load given as part of a standard test meal. 1,25(OH)2D was active even at the lowest dose (0.5 microgram/day), and the slope was such that doubling of absorption would occur at an oral dose of approximately 3 micrograms/day. 25(OH)D was also active in elevating absorption and did so without raising total 1,25(OH)2D levels. On the basis of the dose response curves for 1,25(OH)2D and 25(OH)D, the two compounds exhibited a molar ratio for physiological potency of approximately 100:1. The absorptive effect of vitamin D3 was seen only at the highest dose level (1250 micrograms, or 50,000 IU/day) and was apparently mediated by conversion to 25(OH)D. Analysis of the pooled 25(OH)D data from both the 25(OH)D- and vitamin D3-treated groups suggests that approximately one eighth of circulating vitamin D-like absorptive activity under untreated conditions in winter may reside in 25(OH)D. This is a substantially larger share than has been predicted from studies of in vitro receptor binding.

Absorption↗
PubMed1993

Clinical experience with controlled-release carbidopa/levodopa in Parkinson's disease.

We converted 158 Parkinson's disease (PD) patients on stable doses of standard carbidopa/levodopa (Std-L) to controlled-release carbidopa/levodopa (L-CR). Of the 141 patients who completed the study, 103 (73%) preferred L-CR, 26 (18.5%) preferred Std-L, and 12 (8.5%) had no preference. One hundred fourteen patients elected to continue L-CR, and we performed the primary data analysis on this group. Following conversion to L-CR, patients reported an increase in length of benefit from each dose and an increased "kick-in" time. There was a decrease in the total number of doses, "off" periods, sleep interruptions per night, dose failures, and sleep disturbances. Conversion to L-CR resulted in a significant increase in total levodopa dose. There was no significant change in the dyskinesias. However, early-morning dystonia resolved in eight of 14 patients. Our findings suggest that L-CR is particularly effective in decreasing motor fluctuations, reducing nocturnal problems, and minimizing levodopa dose failures in PD.

Adult↗
PubMed2004

Linguistic styles and complementarities in analyzing character.

This paper discusses and illustrates some technical implications of D. Liberman's contributions to character analysis. Psychoanalysis is an extended conversation, and character structure is expressed linguistically as part of an interactive linguistic field. Liberman's notions of linguistic styles and communicative interaction define a unique analytic surface from which to examine the syntactic, semantic and pragmatic aspects of character. The presence of a rigid and defensive linguistic style reflects resistance and communicative impasse within the transference and countertransference matrix. A fully interactive communicative experience integrates all aspects of the conversation and allows both participants to take part in a shared semantic field, the precondition to the generation of new meanings. Liberman suggests responding to miscommunication by means of a methodical linguistic attitude. Various such complementary responses are explored through several vignettes, with a particular focus on the interplay of the epic and narrative modes.

Character↗
PubMed1999

An analysis of the communication tactics used by hearing-impaired adults.

The aim of this study was to establish whether and to what degree certain types of communication strategies (hearing tactics) used by hearing-impaired adults could be shown to co-occur, and to find out which strategies were more likely to be used in which type of communicative situations. A consecutive series of 100 patients attending an audiological rehabilitation clinic was given a questionnaire asking how often they used each of five different hearing tactics in 11 different situations. 'Avoidance' and 'request for repetition' were the tactics used most commonly. 'Pretending to hear/understand' and 'positioning self to improve hearing' were used less frequently, with 'interruption' the least commonly used. There was some association between tactics and situations but no clear picture emerged. In a reassessment of our methodology and results, we suggest that the future research of communication strategies would benefit from a sociolinguistic approach based on the qualitative analysis of naturally occurring discourse (conversation) of hard-of-hearing people, focusing on the use of different strategies in relation to communicators' goals in interaction.

Adolescent↗
PubMed1993

811 Charnley hips followed for 3-17 years.

10-12 year survivorship analysis of 811 primary or conversion Charnley arthroplasties showed 89 percent probability of mechanical survival (revision excluding sepsis). The mechanical failure rate was 7 percent, and more stems required revision than sockets. The anterio-posterior radiographs of 630 unrevised hips, with a minimum 3-year follow-up, were evaluated using the CART system of terminology. Cement-bone radiolucency was present in 50 percent of sockets and 18 percent of stems. Substantial loosening was present in 6 percent with 94 percent probability of survival at 10-12 years.

Adolescent↗
PubMed2000

[Feeling of well-being and family contacts in community elderly residents].

The aim of this study is to explore the factors associated with feelings of well-being in 683 community-dwelling elderly residents, aged 65 years and over using a self-administered questionnaire. The feeling of well-being was rated using a single question item constructed by The Japanese Association for Cerebro-cardiovascular Disease control (JACD), and then subjects were divided into two groups according to answer categories: happiness and unhappiness. There was no statistically significant difference in the proportion of subjects with happiness between males and females, or among age groups. Even when interaction among multiple factors were adjusted by logistic regression analysis, a high frequency of conversation with family was statistically significantly related to happiness, and the odds ratio of it was highest among all other factors. The odds ratio of high frequency of conversation with family was higher among the elderly aged 75 and over than among those aged 65-74. These results suggest that the construction of communication between the elderly and their family through conversation could contribute to feelings of well-being in the elderly.

Aged↗
PubMed1984

Biological availability of manganese sources and effects of high dietary manganese on tissue mineral composition of broiler-type chicks.

An experiment was conducted with male broiler-type chicks to study tissue uptake of Mn as a measure of biological availability of Mn sources. A basal corn-soybean meal diet (116 ppm Mn) was supplemented with 0, 1000, 2000, or 4000 ppm Mn as either reagent grade sulfate, carbonate, or monoxide and fed ad libitum for 26 days. No toxic effects were noted as expressed by feed intake, weight gain, feed conversion, hematocrit, hemoglobin, or mortality. Analysis of Mn in tissues revealed a highly linear relationship between liver or bone Mn concentration and dietary Mn for all three sources. Manganese concentration in all tissues increased (P less than .01) as dietary Mn increased. Liver and bone Mn accumulation appeared to be excellent indicators of relative biological availability. On the basis of tissue uptake and solubility tests, MnSO4 X H2O was the most available, followed by MnO and MnCO3, respectively. There were no effects of Mn source or level on concentration of Ca, P, Mg, Cu, Zn, or Fe in tissues studied.

Animals↗
PubMed2000

The tourniquet revisited as an adjunct to lower limb revascularization.

PURPOSE: The purpose of this study was to evaluate the role and efficacy of the tourniquet in lower limb revascularization. METHODS: During a 3-year period, 195 patients underwent 205 infrainguinal reconstruction operations in the lower extremity. These patients underwent bypass with a tourniquet and inflow occlusion (group 1) or bypass without a tourniquet (group 2). The type of infrainguinal reconstruction, tourniquet ischemia time, blood loss, and complications related to tourniquet use were recorded. A subset of patients underwent serial muscle biopsies. Specimens from calf muscle were taken just (1) before application of the tourniquet, (2) before tourniquet release, and (3) once wound closure was initiated. These biopsy specimens were studied by histochemical staining and also analyzed for phosphorylase enzyme, a marker for subcellular ischemia. RESULTS: One hundred eleven patients underwent 117 infrainguinal reconstruction procedures in which the tourniquet and inflow occlusion were used. These patients were matched against 84 patients who underwent 88 infrainguinal reconstructions without the use of the tourniquet. Complete hemostatic control in group 1 was obtained in 108 of the procedures (92%). Eight percent of the procedures required minor additional techniques to obtain complete hemostasis; in two instances, the tourniquet was removed because it did not provide hemostasis. Mean tourniquet time was less than 1 hour for all reconstruction groups. There were no instances of neurologic deficit, thrombosis of distal vessels, or vascular injury that was related to the use of a tourniquet. A comparison of the two groups revealed no differences with regard to overall blood loss (P =.63) or duration of operation (P = 0.60), observations that reflect the complexity of the cases rather than the use or nonuse of a tourniquet. When tourniquet control was used, we noted a definite decrease in the time for the distal dissection, because total vascular control with extensive dissection was unnecessary. Histochemical analysis with phosphorylase revealed a conversion of tissue with active enzyme activity to a low level with tourniquet use (P <.05). CONCLUSION: The use of a tourniquet for lower limb revascularization is safe and effective and improves visualization of the operative field. Less dissection of the target vessels is required. With a combination of the nonuse of clamps and other occluding devices, we project a decrease in host hyperplastic response that will, in turn, impact favorably on patency rates. The possibility exists that early failure may be prevented by avoiding the application of traumatic forces to diseased and brittle or calcified arteries. In this study, tourniquet time had no impact on overall operative procedural time, although certain phases of the operation were clearly shortened and facilitated, particularly in complex and difficult reconstructions. Histochemical changes found in muscle biopsy specimens did not adversely impact patients clinically, but further investigation is required to elucidate subcellular events.

Aged↗
PubMedDate not supplied

Laparoscopic colorectal resection: initial experience in a specialist unit.

The adoption of the laparoscopic approach to colorectal resection has been slow amongst colorectal surgeons principally due to concerns regarding oncological safety. Recent randomized controlled trials have confirmed both the safe and some advantage of this procedure have been performing laparoscopic assisted colorectal resection since 2002 and have now performed over 100 cases on non consecutive and selected patients. We have reviewed our experience with the introduction of this technique. 61 patients were operated on for cancer and 39 for benign disease mainly Crohn's and diverticular disease. Operative time was a median of 128 minutes over the course of study. Conversion rate was 5%. Pathological analysis of the resected specimens in the cancer cases revealed adequate lymph node harvest and margins. No patient had a positive margin and no port site metastasis have been seen. Duration of ileus and length of stay were a median of 0 and 6 days. Post operative morbidity and mortality were comparable to open colorectal surgery with the exception of port site herniation which occurred in 4% of patients. This study suggests that a laparoscopic approach to colorectal resection can be successfully introduced in an Irish hospital setting. The challenge facing Irish surgery is to disseminate this technique in a controlled and safe manner for Irish patients.

Adult↗
PubMed2006

[The production of gastrodin through biotransformation of p-hydroxybenzaldehyde by cell suspension culture of Datura stramonium].

AIM: To investigate the production of p-hydroxymethylphenol-beta-D-glucoside (gastrodin) through biotransformation by plant cell suspension cultures. METHODS: Using cell suspension cultures of Datura stramonium to convert the exogenous p-hydroxybenzaldehyde into gastrodin was conducted and the converted compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and chemical evidence. RESULTS: The conversion procedure of p-hydroxybenzaldehyde into gastrodin by Datura stramonium cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the fermental liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of Datura stramonium was also isolated and identified. Two compounds were also isolated from the cell cultures and they were identified as beta-D-furanoallulose (III) and n-butyloxystyryl-beta-D-pyranoallulose (IV). CONCLUSION: Datura stramonium grown in suspension cultures can convert exogenous p-hydroxybenzaldehyde into the corresponding gastrodin.

Benzaldehydes↗
PubMed1977

[Comparative kinetic studies of Mg2+-activated hydrolysis of tripolyphosphate and pyrophosphate by inorganic pyrophosphatase].

Computer analysis of P3 and pyrophosphate conversion rate dependence on substrate and metal-activator concentrations reveals the identity of kinetic patterns. Dissociation and catalytical constants for the enzyme combinations with two types of metal-substrates complexes, MS and M2S, at pH 9.0 are by one to two orders of magnitude "poorer" for P3 as compared to PPi. Optimal pH value for the hydrolysis of P3 is by 2 units higher than this for the hydrolysis of PPi. pH profiles for the kinetic parameters in the pH range 8.0--9.5 differ considerably for the two substrates, presumably due to the existence of additional catalitically important ionisations in the reaction with P3.

Diphosphates↗
PubMedDate not supplied

Interviewing style and response to family information by family practice residents.

The visits of 50 new patients seeing 20 family practice residents were audiotaped. Analysis of transcripts of all conversation involving family-related content showed that the resident physicians used a high control interviewing style consisting mostly of closed-end questions and declarative statements. They rarely asked open-end questions. After eliciting family information, the residents missed numerous opportunities to express empathy or concern verbally. The physicians seldom connected family information obtained during the visit with the patient's ongoing or potential health problems. Of 12 patients reporting a life-threatening familial illness in a close relative, only one received any relevant patient education. The closed style of interviewing appeared to interfere with the physicians' gathering and responding to family information.

Family Practice↗
PubMed1979

Two-dimensional electrophoretic analyses of proteins synthesized during differentiation of 3T3-L1 preadipocytes.

The changes in protein composition and cell surface proteins that occur during the adipocyte conversion of 3T3-L1 preadipocytes were monitored by two-dimensional polyacrylamide gel electrophoresis folowing incubation of cells with [35S]methionine for periods of 3 and 24 h. Alterations in the biosynthesis of more than 30 cytoplasmic proteins, 9 non-histone, chromosome-associated proteins, and 24 membrane proteins, were detected. Although the methodological limitations of the electrophoretic systems employed result in an underestimate of the total number of differences, the alterations observed exceed the enzyme changes known to occur during differentiation of these cells. One major alteration occurring during differentiation is a decrease in the content of a protein whose position following two-dimensional electrophoresis tentatively identified it as actin. A fall in actin content accompanying adipocyte conversion was confirmed by direct analysis of the DNase 1 inhibitory activity in homogenates prepared from cells during the course of differentiation. Studies of cell surface proteins by lactoperoxidase-catalyzed iodination reveal a number of changes during differentiation including an increase in a polypeptide(s) in the molecular weight range of 16,500 to 18,500, a decrease in at least four proteins of molecular weights greater than 100,000, and in a protein of molecular weight 95,000.

Actins↗
PubMed1996

New data on the rate of progression of impaired glucose tolerance to NIDDM and predicting factors.

The Hoorn study has investigated the prevalence of impaired glucose tolerance (IGT) and diabetes in a random sample of the population, 50-75 years of age, in the small Dutch town of Hoorn. The percentage of the population identified as having IGT was 10.3%, while 8.4% had diabetes. The prevalence of both diabetes and IGT was higher in older subjects (70-74 years of age) than in younger subjects (50-54 years of age), suggesting that age is a major determinant of the prevalence of IGT and diabetes in a population. A prospective study of the development of non-insulin dependent diabetes mellitus (NIDDM) in the subjects identified as having IGT was then initiated. Preliminary results for the 158 subjects who have been followed for a mean of 2 years indicated that 28.5% (95% confidence interval, 15-42%) have progressed to NIDDM within this period. The progression rate calculated from this value is 13.8%/year (95% confidence interval, 3.5-24%/year). Analysis of possible determinants of conversion has revealed that 2-hour plasma glucose levels greater than 9.4 mmol/litre and increased fasting proinsulin levels are predictive of progression to NIDDM. This suggests that beta cell dysfunction, rather than increased insulin resistance, is responsible for the development of NIDDM.

Age Factors↗