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Biomedical subjects

R Weinstein

Publications and source records attributed to R Weinstein.

At least 145 records · Page 8Linked to original sources

Mutagen treatment of single Chinese hamster ovary cells produces colonies mosaic for glucose-6-phosphate dehydrogenase activity.

When populations of single Chinesee hamster ovary (CHO) cells were exposed to the mutagen ethyl methane sulfonate (EMS), allowed to grow into colonies, and stained for glucose-6-phosphate dehydrogenase (G6PD) activity, two types of unstained colonies were observed at a frequency of about on per thousand stained colonies. These negative-staining colonies consisted of (1) colonies uniformly deficient in staining activity (pure); and (2) colonies containing both stained and unstained sectors (mosaic) in various relative sizes and patterns. Unstained cells isolated from mosaic colonies were genetically stable and had significantly reduced or absent G6PD activity. Random cell aggregation or chromosome segregation from tetraploid cells is not a significant cause of the sectoring phenomenon. Also, mosaic colonies are not principally caused by mutation at one of two replicated G6PD genes and their subsequent segregation during division. The simplest explanation for this phenomenon is that EMS induces a mutational change in one of the two DNA strands and DNA replication then produces normal and mutant double-stranded DNAs which segregate into wild-type and G6PD-deficient cell types, producing a mosaic colony.

Animals↗

Vascular smooth muscle cell growth kinetics in vivo in aged rats.

Age is a risk factor in the development of atherosclerosis. In this study we investigated the hypothesis that proliferation of vascular smooth muscle cells (SMCs), an integral part of atherosclerotic plaque formation, changes with age. SMC growth kinetics of old rats (21-24 months) were compared to those of young adult rats (3-4 months). Rat aortas were denuded of their endothelium and the animals were killed after [3H]thymidine and Evans blue injections at 0-28 days after denudation. Incorporation of [3H]thymidine into SMC peaked in the young animals by day 2, whereas the older animals responded to endothelial removal with greater incorporation at day 2 and a more sustained rate of incorporation peaking at day 4. The [3H]thymidine incorporation curves decreased sharply from their peaks at 2 and 4 days, respectively, and paralleled each other after day 7. [3H]Thymidine uptake reflected the subsequent SMC intimal growth as measured morphometrically, with old animals showing greater numbers of intimal SMC than did the younger animals. The difference in response of SMC to injury with age suggests that aging produces a change in the vascular SMC that enhances proliferation. This change in response implies that the more pronounced atherosclerotic plaque growth seen with aging may be a result of an age-related increase in response to injury rather than merely the accumulation of time-related intimal change.

Aging↗

Organizational behavior of human umbilical vein endothelial cells.

Culture conditions that favor rapid multiplication of human umbilical vein endothelial cells (HUV-EC) also support long-term serial propagation of the cells. This is routinely achieved when HUV-EC are grown in Medium 199 (M-199) supplemented with fetal bovine serum (FBS) and endothelial cell growth factor (ECGF), on a human fibronectin (HFN) matrix. The HUV-EC can shift from a proliferative to an organized state when the in vitro conditions are changed from those favoring low density proliferation to those supporting high density survival. When ECGF and HFN are omitted, cultures fail to achieve confluence beyond the first or second passage: the preconfluent cultures organize into tubular structures after 4-6 wk. Some tubes become grossly visible and float in the culture medium, remaining tethered to the plastic dish at either end of the tube. On an ultrastructural level, the tubes consist of cells, held together by junctional complexes, arranged so as to form a lumen. The smallest lumens are formed by one cell folding over to form a junction with itself. The cells contain Weibel-Palade bodies and factor VIII-related antigen. The lumens contain granular, fibrillar and amorphous debris. Predigesting the HFN matrix with trypsin (10 min, 37 degrees C) or plasmin significantly accelerates tube formation. Thrombin and plasminogen activator had no apparent effect. Disruption of the largest tubes with trypsin/EDTA permits the cells to revert to a proliferative state if plated on HFN, in M-199, FBS, and ECGF. These observations indicate that culture conditions that do not favor proliferation permit attainment of a state of nonterminal differentiation (organization) by the endothelial cell. Furthermore, proteolytic modification of the HFN matrix may play an important role in endothelial organization.

Cell Differentiation↗

Protection against experimental pyelonephritis by antibodies to pili.

Bacterial pili have been shown to be an important virulence factor for urinary tract infections. In this report we relate the results of studies which evaluated the influence of antipili antibody on the susceptibility of rats to ascending pyelonephritis and on several antibody-mediated antibacterial mechanisms. Rats immunized with E. coli type 1 pili, and animals infected with E. coli developed antipili antibodies in their serum. Active or passive immunization of rats with pili protected the animals from ascending pyelonephritis. Antipili antibody did not mediate complement-dependent bacteriolysis, opsonophagocytosis or promote more rapid intravascular clearance of injected E. coli. Humoral immunity to pili did, however, effectively inhibit bacterial adherence to epithelial cells. These studies indicate that type 1 E. coli pili are immunogenic and that antipili antibodies afford protection from ascending pyelonephritis. They suggest further that a mechanism of protection is inhibition of bacterial adherence.

Animals↗

Hormonal requirements for growth of arterial smooth muscle cells in vitro: and endocrine approach to atherosclerosis.

In this study the hormonal requirements for the growth of arterial smooth muscle cells in vitro were determined. A serum-free, biochemically defined medium, supplemented with the relevant hormones, permitted proliferation and propagation of normal diploid mammalian arterial smooth muscle cells. Serum-free, hormone-supplemented cultures spontaneously formed atherosclerotic plaque-like nodules. Thus atherosclerosis may be mediated by a complex endocrine system.

Animals↗

An endocrine approach to the control of epidermal growth: serum-free cultivation of human keratinocytes.

Human keratinocytes, derived from the skin of newborns and of adults, were grown in the complete absence of serum, in a hormone-supplemented medium on fibronectin-coated cell culture dishes at low seed density. The cell culture medium consisted of Medium 199 containing epidermal growth factor, triiodothyronine, hydrocortisone, Cohn fraction IV, insulin, transferrin, bovine brain extract, and trace elements. Removal of the brain extract from the hormone supplement had a greater negative impact on proliferation of the keratinocyte cultures than did the removal of epidermal growth factor, hydrocortisone, and triiodothyronine or Cohn fraction IV. The growth of keratinocytes in this hormone-supplemented medium suggests that control of keratinocyte growth depends in large part on endocrine stimulation by other body organs, including the brain.

Brain Chemistry↗

CT scan of the spine for herniated discs.

The high resolution conversion kit introduced for the EMI CT 5005 scanner has twice the spatial resolution of the usual scan mode and has proven to be quite useful in evaluation of the spinal canal. The present study was performed to determine the capability of the high resolution system to diagnose herniated lumbar disc without intrathecal contrast. HNP was diagnosed in 83% of the cases which had a positive myelogram, 70% of which had operation and all were proven positive.

Adult↗

Serial propagation of human endothelial cells in vitro.

Human umbilical vein (HUV) endothelial cells were grown for 15 to 21 passages at a split ratio of 1:5 (at least 27 population doublings) on a human fibronectin (HFN) matrix in Medium 199 supplemented with fetal bovine serum (FBS) and endothelial-cell growth factor (ECGF). This system also permitted the growth of HUV endothelial cells at cell densities as low as 1.25 cells/cm2. In addition to delaying the premature senescence of HUV endothelial cells, ECGF also reduced the serum requirement for low-density HUV endothelial-cell growth; 2.5% serum and ECGF yields half-maximum growth as compared to high serum controls. Significant HUV endothelial-cell growth was also observed in medium supplemented with either ovine hypophysectomized (HYPOX) serum, plasma-derived serum (PDS), or HYPOX-PDS in the presence of ECGF, suggesting that neither the pituitary nor the platelet contributes to HUV endothelial-cell growth.

Animals↗

The morphological and biochemical characterization of a line of rat promegakaryoblasts.

Biochemical and morphological evidence is presented to support the characterization of a rat bone-marrow-derived cell line (RPM) as an analog of the promegakaryoblast. The conditions for in vitro growth and maturation of the RPM line are described. Rapid proliferation of the RPM line is readily achieved when cultures are supplemented with moderate levels of fetal bovine serum (FBS). The proliferative compartment is a small blast-like cell. Immunofluorescent staining demonstrates that the RPM cells contain factor VIII:antigen and fibrinogen in their cytoplasm. The cells secrete, into their conditioned medium, a potent mitogenic activity for rat aortic smooth muscle cells. When incubated under conditions of relative serum deprivation, the cells stop proliferating and undergo a process of maturation. The sequence of maturation is described as stage I (promegakaryoblast--the proliferative compartment); stage II (immature megakaryocyte or promegakaryocyte); stage III (mature megakaryocyte). The stage III cells release, from their cytoplasm, small membrane-bound vesicular bodies containing lavender granules and cytoplasmic organelles. These have been designated stage IV. The RPM line may provide a useful model for the in vitro study of megakaryocyte maturation and the synthesis of megakaryocyte-specific proteins.

Animals↗

Effects of cerulenin on antibiotic-induced lysis of streptococcus faecalis (S. faecium).

Addition of the antibiotic cerulenin to cultures lowered the minimal effective concentration of penicillin G or methicillin required to produce bacterial lysis and killing. This effect was most pronounced at subinhibitory antibiotic concentrations. Cerulenin had no significant effects on lysis or killing induced in the presence of D-cycloserine, fosfomycin, bacitracin, or vancomycin.

Anti-Bacterial Agents↗

Immuno-heterogeneity of the calcitonins of hypercalcemia, breast and lung cancers and medullary carcinoma of thyroid.

Four antisera raised in the goat have very different properties: all recognized the immunoreactive calcitonin (iCT) of medullary carcinoma of the thyroid (MCT), one the response of normal subjects to induced or endogenous hypercalcemia and 2 others a different molecular species which occurs in half the patients with cancer of breast and 3/4 of patients with cancer of the lung. The latter two antisera are most sensitive to the 22-32 sequence of human calcitonin. Depending on the antiserum used, 4 or 7 peaks of immunoreactivity are found in eluates by column chromatography or stimulated serum from MCT. Not all elevated levels of iCT in serum are diagnostic of MCT and ectopic production by lung and breast cancer must be considered. Presence of higher levels of iCT with greater amounts of cancer tissue and undetectable levels after surgery or radiotherapy when using antisera which require intact molecule of calcitonin for recognition suggest the possibility that sequential calcitonin levels with differentiating antisera may be helpful in assessing the extent of disease and response to therapy.

Breast Neoplasms↗

Phagocytic resistance of Escherichia coli K-1 isolates and relationship to virulence.

Blood culture isolates from 133 episodes of Escherichia coli bacteremia were typed for K-1 capsular antigen by immunodiffusion, utilizing equine antiserum raised against meningococcal group B polysaccharide. Twenty-six percent (34 of 133) of these isolates were positive for K-1 antigen. These 133 strains, 34 K-1 and 99 non-K-1, were tested for susceptibility to phagocytosis. K-1 strains were found to be more resistant to clearance (27%) than non-K-1 strains (71%) when tested in an in vitro opsonophagocytic/killing assay containing normal human granulocytes and plasma. Additional studies demonstrated that resistance was due to decreased phagocytosis rather than diminished intraleukocytic killing. K-1 strains obtained from stool showed a similar degree of resistance to phagocytosis when compared with K-1 blood isolates. A comparison of clinical data on non-neonatal patients with E. coli K-1 and non-K-1 bacteremia showed no significant differences in mortality for these two groups. The incidence of shock for patients bacteremic with K-1 strains (74%) was significantly greater than that for patients bacteremic with non-K-1 strains (33%). These differences are attributed to the increased resistance to phagocytosis observed for K-1 versus non-K-1 E. coli isolates.

Adult↗

Attenuation values in computed tomography of the abdomen.

Attenuation values were obtained from 50 CT examinations of the upper abdomen. A normal range of values, obtained for the organs in the upper abdomen, was compared with patient size and age. Rank ordering according to the mean attenuation values was also performed. The most significant finding was that the liver normally had the highest attenuation value of any of the viscera measured. When another organ in the upper abdomen had a mean value greater than that of the liver, this reflected severe systemic disease.

Humans↗