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Biomedical subjects

J Margolis

Publications and source records attributed to J Margolis.

At least 37 records · Page 2Linked to original sources

Identification and behavior of epithelial stem cells in the Drosophila ovary.

Throughout their lives, adult Drosophila females continuously produce oocytes, each surrounded by an epithelial monolayer of follicle cells. To characterize the somatic stem cells that give rise to ovarian follicle cells, we marked dividing cells using FLP-catalyzed mitotic recombination and analyzed the resulting clones. Each ovariole in young females contains, on average, two somatic stem cells located near the border of germarium regions 2a and 2b. The somatic stem cells do not coordinate their divisions either with each other or with the germline stem cells. As females age, initially mosaic ovarioles become monoclonal, indicating that functional somatic stem cells have a finite life span. Analysis of agametic flies revealed that somatic cells continue to divide in the absence of a germline. Under these conditions, the somatic stem cells develop near the tip of the ovariole (the normal site of the germline stem cells), and a subpopulation of somatic cells that normally separates the germline and somatic stem cells is missing.

Animals↗

Six month outcome and determinants of adverse clinical events after successful excimer laser coronary angioplasty. ELCA A.I.S. Multicenter Registry.

OBJECTIVE: To describe the six month outcome and predictors of adverse clinical events following successful excimer laser coronary angioplasty (ELCA). DESIGN: Retrospective analysis of comprehensive data collected at time of initial procedure and during planned follow-up intervals. SETTING: There were 35 participating institutions throughout the United States. PATIENTS: The study population was comprised of 3,069 patients who had successful ELCA and completed follow-up at 6 months, representing 91% of eligible cases. Forty percent had prior balloon angioplasty and 34% had prior bypass surgery. MEASUREMENTS: Patient symptomatology was assessed by Canadian Cardiovascular Society functional (CCSF) class. Interim adverse clinical events [death, Q wave myocardial infarction (QMI), repeat intervention, bypass surgery, and a composite end-point] were tracked. Twenty-six pre-ELCA clinical, angiographic and procedural variables were evaluated to assess their ability to predict outcome. RESULTS: At 6 months, there was 2.8% mortality, 2.5% incidence of QMI, and 28.7% required either repeat intervention or bypass surgery. Overall, 69.2% of patients had no adverse event. In this cohort, the pre-ELCA CCSF class grade was 2.7 +/- 1.2, but at 6 months, it was 0.7 +/- 1.1 (p < 0.001). The multivariate predictors of the composite end point were gender, stenosis location, and rest and unstable angina. CONCLUSIONS: There is an excellent 6 month outcome after successful ELCA in this selected population with complex coronary artery disease. The majority of patients without interim events enjoy significant alleviation of symptoms. The predictors of adverse events are clinical rather than angiographic.

Angioplasty, Balloon, Laser-Assisted↗

Multifunctional apparatus for electrokinetic processing of proteins.

New equipment (the "Gradiflow") has been designed and constructed to provide efficient large-scale preparative fractionation of macromolecules, based on charge and/or size differences, as well as the concentration of macromolecules and electrodialysis. Examples of its capability are the separation of a mixture of haemoglobin (50 mg) from bovine serum albumin (50 mg) within 15 min (based on charge differences at pH 6.8), the purification of phycoerythrin from a crude extract on the basis of size, and the fractionation of serum proteins into two discrete size classes.

Animals↗

Percutaneous excimer laser coronary angioplasty: results in the first consecutive 3,000 patients. The ELCA Investigators.

OBJECTIVES: We report the comprehensive results of the first consecutive 3,000 patients treated in an excimer laser coronary angioplasty registry. BACKGROUND: Excimer laser coronary angioplasty involves the use of a pulsed, 308-nm ultraviolet laser transmitted by optical fibers to reduce coronary stenoses. Preliminary reports have described safety and efficacy profiles in small numbers of patients. METHODS: Patients were enrolled in a prospective, nonrandomized manner. The catheters used were 1.3, 1.6, 2.0, 2.2 and 2.4 mm in diameter, at energy densities up to 70 mJ/mm2. Procedures were performed by standard angioplasty technique with conventional guide catheters. RESULTS: Seventy-five percent of patients were male, 68% were in Canadian Cardiovascular Society functional class III or IV and the cohort included 3,592 lesions. Procedural success (final stenosis < or = 50% without in-hospital Q wave myocardial infarction, coronary artery bypass surgery or death) was 90% and did not differ between the first 2,000 and the last 1,000 patients treated. There was no significant difference in success or complication rates with respect to lesion length, nor were there differences between selected complex and simple lesions. Complications included in-hospital bypass surgery (3.8%), Q wave myocardial infarction (2.1%) and death (0.5%). Coronary artery perforation occurred in 1.2% of patients (1% of lesions) but significantly decreased to 0.4% in the last 1,000 patients (0.3% of lesions). Angiographic dissection occurred in 13% of lesions, transient occlusion in 3.4% and sustained occlusion in 3.1%. Comprehensive lesion morphologic data collected in the latter portion of the study showed the procedure predominantly limited to American College of Cardiology-American Heart Association type B2 and C lesions, with no significant difference in short-term outcome between groups. CONCLUSIONS: Excimer laser angioplasty can be safely and effectively applied, even in a variety of complex lesions not well suited for percutaneous transluminal coronary angioplasty. These types may include aorto-ostial, long lesions, total occlusions crossable with a wire, diffuse disease and vein grafts. Most recent data show a trend for the selection of predominantly complex lesions and a reduction in the incidence of perforation. This procedure may broaden the therapeutic window for the interventional treatment of selected complex coronary artery disease.

Angioplasty, Balloon, Coronary↗

Excimer laser coronary angioplasty of aorto-ostial stenoses. Results of the excimer laser coronary angioplasty (ELCA) registry in the first 200 patients.

BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) of aorto-ostial stenosis has been associated with a lower rate of acute success, a high risk of vessel closure, and late restenosis. The purpose of this report is to document a prospective multicenter trial of excimer laser coronary angioplasty (ELCA) of aorto-ostial stenosis involving the coronary arteries and saphenous vein grafts. METHODS AND RESULTS: Between December 1989 and May 1992, 206 aorto-ostial ELCA procedures were performed on 209 stenoses in 200 patients. Canadian Cardiovascular Society class III or IV angina was present in 76%. The distribution of stenosis locations was left main coronary (LM) in 26 (12%), right coronary (RCA) in 124 (59%), and vein grafts (VG) in 59 (28%). Adjunctive PTCA was performed in 72%. Procedure success defined as < or = 50% diameter stenosis without major complications was achieved in 90% (LM, 92%; RCA, 89%, VG, 90%). Quantitative angiographic analysis documented an improvement in stenosis diameter from 0.8 +/- 0.5 mm or 76 +/- 14% at baseline to 2.1 +/- 0.6 mm or 36 +/- 15% at completion (P < .01). The majority of the acute gain in diameter (1.0 +/- 0.6 mm) resulted from ELCA. A major complication during hospitalization occurred in 3.9% (death, 0%; Q-wave myocardial infarction, 0.5%; bypass surgery, 3.4%). The only logistic regression univariate and multivariate predictor of procedure failure was female gender. Six-month angiographic follow-up, available in 51% of eligible patients, documented an average lumen diameter of 1.7 +/- 1.0 mm and mean diameter stenosis of 46 +/- 26%. Restenosis (> 50% diameter stenosis) occurred in 39% (LM, 64%; RCA, 35%; VG, 35%). Restenosis was less likely when residual stenosis was < or = 35% (28% versus 53%, P < .05). Clinical events at follow-up were death, 2.7%; bypass surgery, 6.5%; myocardial infarction, 2.2%; and repeat angioplasty, 16.2%. Of the remainder, 78% were asymptomatic, class I or II for anginal symptoms. An adverse event during follow-up was more than twice as likely in the group with LM (50.0% versus 21.1%, P < .02). CONCLUSIONS: ELCA is acutely effective and safe therapy in patients with aorto-ostial stenosis. Six-month restenosis, adverse-event rates were higher and functional status was poorer in the group with LM stenosis. ELCA may be considered as an alternative to bypass surgery in carefully selected patients with isolated aorto-ostial stenosis of the RCA and saphenous vein grafts.

Adult↗

Rapid and automated characterisation of seed genotype using Micrograd electrophoresis and pattern-matching software.

New precast microgels are described for use in quickly identifying seed of cereal varieties by determining protein composition within an hour. For example, gliadin proteins are extracted from crushed wheat grain, wheatmeal or flour with ethylene glycol (centrifugation not necessary) and 5 microliters extract is applied to a Micrograd gel (3-15% gel gradient) for ten minutes' electrophoresis at 300 volts in sodium lactate buffer (pH 3.1). Alternatively, precast gels are available for SDS gel electrophoresis for examining a different aspect of grain composition as a means of identification. To further expedite identification, software packages have been developed to match the protein pattern for an unknown sample against those of authentic samples, thus to provide quick and definite identity, based on electrophoretic banding, densitometer scan, HPLC profile, multiple antibody reaction or RFLP pattern (PatMatch program). Furthermore, the program WhatWheat offers advice on the best combination of methods to use for a specific task of identification.

Automation↗

Rapid (ten-minute) pore-gradient electrophoresis of proteins and peptides in Micrograd gels.

Precast gradient gels of short migration length (25 mm) have been developed to provide rapid electrophoretic separation without loss of resolution. These Micrograd gels have been prepared in gel ranges (conventional and unique) to match pore-gradient electrophoresis conditions to proteins/peptides ranging in size from several hundreds to millions. The Hylinx Micrograd gel combines an extreme gel range (6 to 48% polyacrylamide) with a novel crosslinker to provide sieving of polypeptides, and pore-limit electrophoresis of the smallest proteins (e.g. insulin monomer). All gel ranges (such as 3 to 30%) provide zone sharpening in routine analysis of conventional protein mixtures (e.g. serum) within 10 min electrophoresis at 200 to 300 volts. The gels are thin (1 mm) and thus stain quickly, but the gel cassette is of conventional overall width (83 mm), thus fitting many apparatus designs and accommodating 12 samples. The gels are finding valuable use in screening applications, requiring the electrophoretic analysis of many samples, and in cases where a rapid answer is needed, such as monitoring protein purification. The gels have proved particularly useful, in-house, for the latter application in developing Gradipore's new large-scale preparative electrophoresis system, the Gradiflow.

Electrophoresis, Polyacrylamide Gel↗

Prolonged beneficial effects of dobutamine in refractory heart failure--case histories.

Two patients with refractory congestive heart failure were treated with dobutamine for forty-eight to seventy-two hours with close monitoring of weight and symptomatology. After two to four cycles of dobutamine treatment, 1 patient showed sustained hemodynamic benefit after six months, whereas the second patient had partial improvement. At the end of a one-year follow-up, both had sustained improvement in effort tolerance, documented by a stress test. The quality of life has markedly improved, and they have not required further hospital care.

Dobutamine↗

Freeze-dried activated substrate for factor VIII assays.

Factor VIII-deficient plasma (natural or artificial) mixed with kaolin and phospholipid can be lyophilized to provide ready-to-use substrate which is stable for months at 4 degrees C and usable after many weeks at room temperature. Factor VIII assays are much simplified and more reproducible using this reagent and can be quantified with the aid of a programmable calculator according to the equation (formula; see text) as % of standard and X, S and B are clotting times of test, standard and blank samples respectively. The slope of the log/log function (k) is approximately--6.5.

Blood Coagulation Tests↗

The telephone management of gastroenteritis by family medicine residents.

In order to describe the telephone management of gastroenteritis by family practice residents, audiotapes and transcripts of telephone calls by 31 family practice residents were analysed with respect to clinical content, temporal patterns, and physicians' utterance form. The study hypothesis were that year 3 residents would have shorter calls, take more complete histories, and score better on other performance measures than year 1 residents, but no significant differences were demonstrated in the study. The mean duration of telephone calls was 4.6 minutes. Residents talked three times longer than the caller. History taking completeness was highly correlated with telephone call duration and particularly with time spent talking by the caller. Significant questions, such as asking about hydration status, were frequently omitted. Most questions were closed (84 percent). Time spent talking by the caller was longer, and more listener feedback was given, with year 1 than with year 3 residents.

Child, Preschool↗

Preparation of stable lyophilized cryoprecipitate in the original frozen plasma bags.

A procedure for preparing single units of freeze-dried cryoprecipitate (cryo) in the original bags is described in detail. Batches of frozen plasma bags were rapidly thawed by rocking in a water bath, centrifuged and after complete removal of supernate the compact cryo pellets (2-5 g) were lyophilized with the aid of a holding tray with a filtered air chamber designed for this purpose. Adding synthetic amino acids has greatly improved stability of the dried concentrate during storage and has increased its resistance to heat denaturation (e.g. less than 10% loss of potency after 20 h at 70 degrees C). The method is presented as a viable option for safe and economical manufacture of a stable factor VIII concentrate.

Blood Preservation↗

Human placental alkaline phosphatase in liver and intestine.

Three distinct forms of human alkaline phosphatase, presumably isozymes, are known, each apparently associated with a specific tissue. These are placental, intestinal, and liver (kidney and bone). We have used a specific immunoassay and HPLC to show that placental alkaline phosphatase is also present in extracts of liver and intestine in appreciable amounts.

Alkaline Phosphatase↗

A process for preparation of 'high-purity' factor VIII by controlled pore glass treatment.

A simple process for large-scale manufacture of 'high-purity' factor VIII is described in detail. A crude concentrate prepared from washed cryo is treated with controlled pore glass (CPG, 500 A pore diameter) in proportion of 20-30 ml of CPG to 1 g input of protein. The slurry is poured into a separation column and the effluent purified concentrate collected. The remaining factor VIII in the void volume is displaced by a wash solution. After passage through a 0.2 micron membrane filter the product is dispensed and lyophilized. Maintaining the operating pH at 6.5-6.7 and adding synthetic amino acids improved the yield and solubility. The current concentrate contains 1 unit of factor VIII per mg protein (10 units mg fibrinogen) with a recovery of 250 units/kg plasma. The CPG stage is non-destructive, yielding more than 90% of the input factor VIII. In 1980-1983, more than 3 X 10(6) units have been used in New South Wales, mostly for massive cover in surgical patients. In collaboration with the Commonwealth Serum Laboratories, it is intended to expand production for use in other Australian States.

Amino Acids↗

Electrophoresis of small proteins in highly concentrated and crosslinked polyacrylamide gradient gels.

A high concentration (40%) of acrylamide plus N,N'-methylenebisacrylamide combined with a high level of crosslinking (12.5%) yielded clear gels capable of restricting the passage of small proteins. This gel composition was chosen in preference to other combinations, in particular those producing opaque gels which have larger pore sizes and which provide a reduced sieving effect. Gradient gels were prepared in which the gel concentration rose from 3 to 40% and the degree of crosslinking increased from 4 to 12.5%. Such gels were suitable for fractionating crude, unreduced, and uncharacterized extracts containing proteins ranging in molecular size from 10,000 to several million daltons under conditions where all proteins are retained on the gel even after prolonged electrophoresis. The gels yielded zones which were of improved sharpness and resolution compared with gels of lower concentration and degree of crosslinking, and can be used to provide an estimate of molecular size. Examples of the use of HX gradient gels included both anodic and cathodic electrophoresis at pH 8.3 and 3.1, respectively, of serum and cereal-grain proteins and a partial enzymic hydrolysate of serum albumin.

Acrylamides↗