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Biomedical subjects

J Feder

Publications and source records attributed to J Feder.

At least 91 records · Page 5Linked to original sources

Synthetic substrates of vertebrate collagenase.

The active site specificity of vertebrate collagenase was mapped with the synthesis of a variety of peptides, peptolides, and peptide esters. The enzyme was found to prefer very lipophilic sequences, and it was also found to be an esterase. The thio peptolide Ac-Pro-Leu-Gly-SCH[CH2CH(CH3)2]CO-Leu-Gly-OC2H5 was found to be an exceptional substrate. High-performance liquid chromatography and tandem mass spectrometry were used to unambiguously establish the cleavage site in several peptide substrates.

Animals↗

Fibrin-enhanced endothelial cell organization.

The formation of cloned bovine endothelial cells into capillary-like tubes is accelerated from 3-7 days to 2-18 h in the presence of fibrin. Indirect immunofluorescence showed the presence of both fibrin and fibronectin in the strands along which the cells organized. Electronmicroscopy revealed the same type of cell structures as form in the absence of fibrin; it also revealed a gradual decrease with time of the fibrin within the putative lumen. Fibrin and fibronectin are commonly present during angiogenesis in vivo, thus these in vitro observations may well have relevance to the in vivo process.

Agar↗

Spectrophotometric assay for vertebrate collagenase.

Collagenase from normal human skin fibroblasts was found to catalyze the hydrolysis of esters and thio esters. This observation led to the development of a rapid, sensitive, continuous spectrophotometric assay for vertebrate collagenase using the thio peptolide Ac-ProLeuGly-S-LeuLeuGly-OC2H5 as substrate in the presence of 4,4'-dithiodipyridine or Ellman's Reagent. A Km of 0.004 M and a kcat of 370,000 h-1 were determined for the thio peptolide-enzyme reaction. The method is able to detect collagenase at concentrations as low as 2 ng/ml.

Fibroblasts↗

Close linkage of alpha and beta interferons and infrequent duplication of beta interferon in humans.

Five restriction fragment length polymorphisms in the human alpha/beta interferon (IFN-alpha/beta) gene region were identified, three with an IFN-alpha probe and two with an IFN-beta probe. Heterozygosities are 74% for IFN-alpha, 57% for IFN-beta, and 87% jointly, making IFN-alpha/beta genes excellent markers for the short arm of chromosome 9. The pedigrees of about 25 families of Caucasian background were studied. Segregation analysis disclosed the occurrence of 12 of 32 possible haplotypes. No recombinant was found between IFN-alpha and -beta genetic markers; linkage disequilibrium within the IFN-alpha markers is of a similar order of magnitude as that between the IFN-alpha and -beta markers. The IFN-alpha and -beta genes might cluster within several hundred kilobases. In two parents, the IFN-beta gene is duplicated; the duplications segregate regularly. They are of independent, probably recent, origin. Therefore, some degree of multiplicity might even be found for IFN-beta, at least in some individuals.

Adult↗

A discordant sibship analysis between beta-NGF and neurofibromatosis.

A new restriction fragment length polymorphism 5' to the beta-nerve growth factor (beta-NGF) gene has been found in proximity to the BglII polymorphism, and both polymorphisms are detectable with an EcoRI 7-kilobase (kb) subclone. Absence of the TaqI recognition site lengthens the 4.3-kb and 1.7-kb hybridizing fragments to 6 kb, and the alleles are in Hardy-Weinberg equilibrium with frequencies of 83% and 17%, respectively. Previous research has suggested that NGF is involved in disseminated neurofibromatosis (NF). We found four informative disseminated NF families with the two beta-NGF polymorphisms and have provided clearcut evidence against beta-NGF gene alteration in these families. If disseminated NF is found to be heterogeneous at a molecular level, more families should be tested to further rule out any role for beta-NGF in this syndrome.

Base Sequence↗

DNA restriction fragment analysis of the proopiomelanocortin gene in schizophrenia and bipolar disorders.

The method of DNA restriction fragment analysis using gene probes for the proopiomelanocortin (POMC) gene was employed to detect possible molecular variation in the POMC gene in schizophrenia and bipolar illness. No gross structural abnormalities in restriction fragments were observed with the set of restriction enzymes used. Two allelic restriction sites were observed giving rise to fragment length polymorphisms. One of these is a new polymorphism, not previously reported, which will be of value as a linkage marker. The associations between the two DNA polymorphisms that are closely linked to the POMC gene and both schizophrenia and bipolar disorder were investigated. No association was found, thus adding weight to the evidence that there are no alterations in the POMC gene in schizophrenia and bipolar illness.

Base Sequence↗

A systematic approach for detecting high-frequency restriction fragment length polymorphisms using large genomic probes.

Thirteen phage clones containing low-copy sequences were isolated from a human DNA library and tested for their ability to detect restriction fragment length polymorphisms (RFLPs). Reported are the RFLPs revealed with each clone, all found in frequencies useful for linkage studies. Cytological data are available for five of the 13 clones, with regional assignments made for three of the markers by in situ hybridization. It is concluded that phage clones containing large unique DNA inserts detect multiple RFLPs with high efficiency. An analysis of the relative efficiency of 20 restriction enzymes for detecting single nucleotide changes is discussed by comparing the observed data to those expected on the basis of recognition and potential site frequencies, as computed from the dinucleotide distribution. Finally, in an effort to facilitate linkage studies using polymorphic DNA sequences, experiments were made with pools of probes from various sources.

Bacteriophages↗

Mammalian cell culture: technology and physiology.

During the last decade phenomenal advances have taken place in large-scale mammalian cell culture both for microcarriers and suspension methods. The cost of serum and product quality require that such systems be examined in terms of both their physical and chemical parameters. Data are presented as to the method used to more than double the final harvest cell density of 100-liter batch culture reactors by simple temperature measurements and 12-liter reactors by increasing the oxygen potential of the liquid. The results are interpreted in terms of physical transport phenomena of momentum, heat and mass via the theoretical relationships of Navier -Stokes, Fourier and Fick respectively. In addition, data are presented of 3-, 12-, and 100 liter reactors that the spent cell growth media were analyzed for ten different chemistries. The Monod cell growth curve was used to interpret the results. The cost savings and product quality improvements can be enormous by approaching both the technology and physiology of mammalian cell culture from the vantage points of chemistry, thermodynamics and transport phenomena.

Animals↗

Poor people and poor hospitals: implications for public policy.

In 1980, while most hospitals were in reasonably good financial health, hospitals heavily involved in serving the poor ran a considerable risk of financial trouble. Fewer than 9 percent of the nation's hospitals accounted for 40 percent of the nation's total care to the poor. These hospitals, almost half of which were in the 100 largest cities, not only devoted more of their care to the poor than other hospitals, they also served substantially smaller proportions of privately-insured patients. The result was that one-third of these hospitals--by themselves accounting for over 15 percent of all care to the poor--ran deficits in 1980. Using data from a 1980 survey of nonfederal, nonprofit hospitals, this paper examines the fiscal situation of hospitals heavily involved in serving the poor. The analysis shows that it is insufficient revenues, not inefficiency or underuse, that creates these hospitals' financial problems. The article concludes with an assessment of several policies that could be adopted to alleviate this financial pressure and sustain care to the poor.

Costs and Cost Analysis↗

Falling through the cracks: poverty, insurance coverage, and hospital care for the poor, 1980 and 1982.

About one-third of the nation's poor lack insurance--public or private--against the costs of illness. Data from 1980 and 1982 show that a patchwork of state and local government charitable grants, and the disparate efforts of hospitals to provide free care, cannot mend the national "safety net." A prudent short-run approach to modifying charity care is advanced, although the long-run necessity for insuring the uninsured is inevitable.

Charities↗

Left ventricular volume and ejection fraction response to exercise in aortic regurgitation.

To test the hypothesis that left ventricular (LV) performance in aortic regurgitation (AR) can be more completely characterized by measurement of LV volumes in addition to ejection fraction (EF), 27 asymptomatic patients (Group 1), and 22 symptomatic patients (Group 2), and 10 control subjects were studied at rest and during upright bicycle exercise using the first-pass technique and a multicrystal scintillation camera. LV end-diastolic volume was measured by the area-length method. In the control group end-diastolic volume increased 14%, end-systolic volume decreased 22%, and EF increased 22% with exercise. In contrast, in Group 1 patients with AR, end-diastolic volume was elevated at rest and during exercise. The 18% decrease in end-diastolic volume during exercise was significantly different from the control response (p less than 0.01). End-systolic volume was also elevated at rest and during exercise, but the 30% decrease during exercise was a response not significantly different from the control. Although mean EF increased 15% in these patients, EF at peak exercise was significantly lower than that in the controls. In Group 2 patients with AR, resting EF was reduced, the EF response to exercise was abnormal, and end-diastolic and end-systolic volume responses to exercise were significantly different from those in Group 1: end-diastolic volume did not change and end-systolic volume increased. In contrast to the fairly uniform volume responses among all Group 1 patients, there were 2 subgroups based on volume changes within Group 2: 7 of 22 had a decrease in end-diastolic volume and end-systolic volume during exercise and 8 of 22 showed an increase in end-diastolic and end-systolic volume during exercise. In conclusion, LV volumes at rest and exercise give more information about LV functional reserve in symptomatic patients with AR than do EF responses alone, and may be useful in separating symptomatic patients who show a normal end-systolic volume response to exercise from those in whom worsening failure develops during exercise.

Aortic Valve Insufficiency↗

The formation of capillary-like tubes by calf aortic endothelial cells grown in vitro.

Cloned, large vessel endothelial cells derived from fetal bovine and bovine calf aortas formed three-dimensional structures in vitro without tumor-conditioned medium or special substrata. Transmission electron microscopy showed the structures to be hollow tubes composed of typical endothelial cells with overlapping and interdigitating cytoplasmic processes typical of those seen in in vivo capillaries. The putative lumen of these tubes generally contained abundant electron-dense fibrous material, which by ruthenium red and indirect immunofluorescent staining appeared to be extracellular matrix. This suggests that the endothelial cell orientation in the tubes is the reverse of that normally found in in vivo vessels.

Animals↗

Effect of interferon on the penetration of murine leukemia virus and the binding of its genome RNA to polyribosomes at the early stage of infection.

The effect of interferon (IFN) on the adsorption, penetration and subsequent binding of the incoming genome RNA of Moloney murine leukemia virus (MLV) to polyribosomes, was studied in NIH/3T3 mouse fibroblasts. Virus adsorption was assayed by determining reverse transcriptase activity in the inoculating virus stock and in the cell membrane fraction before and after 45 minutes of infection. Both measurements suggested that IFN had no effect on virus adsorption. Virus penetration was determined by measuring the amount of viral RNA in the cell cytoplasm at 45 minutes after infection. This amount was remarkably lower in IFN-treated than in untreated cells. This reduction was not due to inhibition of a possible induction of endogenous viral genetic information by the penetrating virions, but was proved to be a direct effect of IFN on virus penetration, which was related to the IFN-induced antiviral state. The effect of IFN on binding of parental genome RNA to polyribosomes was then investigated by analysing Crt hybridization kinetics of polyribosomal viral RNA at different time intervals after infection. While in untreated control cells maximal binding occurred at 3 hours postinfection, this maximal binding was observed in IFN-treated cells at 5 hours postinfection. The distribution of viral RNA molecules between sub-cytoplasmic fractions at 3 hours after infection was, in IFN-treated cells, significantly different from that observed in the untreated cells.

Adsorption↗