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Biomedical subjects

C J Jones

Publications and source records attributed to C J Jones.

At least 163 records · Page 9Linked to original sources

Characterization of the fliE genes of Escherichia coli and Salmonella typhimurium and identification of the FliE protein as a component of the flagellar hook-basal body complex.

Within flagellar region III of Escherichia coli and Salmonella typhimurium, the genomic organization has been largely established. An exception is fliE, a gene whose exact location and product function are not well understood. We cloned the fliE gene, obtained its DNA sequence, and identified its product.fliE was found to be a monocistronic transcriptional unit, adjacent to and divergent from the large fliF operon. It is several kilobases distant from the nearest flagellar operon in the other direction, the fliD operon, and constitutes the first operon within the newly defined region IIIb, which contains the genes fliE through fliR.fliE encodes a small, moderately hydrophilic protein with a deduced molecular mass of 11,114 Da (E. coli) or 11,065 Da (S. typhimurium). We identified a protein within the isolated hook-basal body complex as the fliE gene product on the basis of its size and comparison of its N-terminal amino acid sequence with that deduced from the gene sequence. From gel electrophoresis and autoradiography of 35S-labeled S. typhimurium hook-basal body complexes (C.J. Jones, R.M. Macnab, H. Okino, and S.-I. Aizawa, J. Mol. Biol. 212:377-387, 1990) and the deduced number of sulfur-containing residues in FliE, we estimated the stoichiometry of the protein in the hook-basal body complex to be about nine subunits. FliE does not undergo cleavage of a signal peptide, nor does it show any sequence similarity to the axial components like the rod or hook proteins, which are believed to be exported by the flagellum-specific export pathway. On the basis of this and other evidence, we suggest that FliE may be in the vicinity of the MS ring, perhaps acting as an adaptor protein between the ring and rod substructures.

Amino Acid Sequence↗

Localization of cis-acting sequences in the latency-related promoter of bovine herpesvirus 1 which are regulated by neuronal cell type factors and immediate-early genes.

Bovine herpesvirus 1 (BHV-1) establishes a latent infection in sensory ganglionic neurons of cattle. During a latent infection, a single latency-related (LR) transcript is expressed. This observation suggested that DNA sequences in the LR promoter are positively regulated by neural cell type factors. The regulation of the LR gene was examined in neural cells as well as nonneural cells in transient assays. A 258-bp XbaI-SphI fragment from the LR promoter cis activated the herpes simplex virus type 1 thymidine kinase promoter in rat pheochromocytoma (PC-12) cells and differentiated human (HCN1A) neurons. In contrast, cis activation was not observed with rat (Rat-2) fibroblasts, undifferentiated HCN1A cells, or bovine turbinate cells. Treatment of PC-12 cells with nerve growth factor increased transcriptional activity of the XbaI-SphI fragment. Exonuclease III footprinting experiments suggested that nuclear factors bind to the XbaI-SphI fragment. The immediate-early genes of BHV-1 trans activated the LR promoter, and DNA sequences 5' to the XbaI-SphI fragment were necessary for maximal stimulation. These results imply that neural-cell-type-specific factors and BHV-1 immediate-early genes positively regulate LR gene expression.

Animals↗

Blood velocity distributions within intact canine arterial bifurcations.

As local variations in blood flow are implicated in atherogenesis at bifurcations, we measured in vivo blood velocities in different planes within exposed iliofemoral arterial bifurcations in 8 dogs using 20-MHz, 80-channel Doppler ultrasound velocimetry. Cardiac frequency was fixed at 2 Hz by pacing. Local geometry was characterized using 25-MHz, B-mode ultrasound images, photographs, and methacrylate casts. The bifurcations were asymmetrical and planar to within 5 degrees, the diameter ratios of the daughter vessels ranged from 1.47 to 2.00, and the angles between them ranged from 40 to 76 degrees. Measured velocities indicated that just upstream of the bifurcation mean peak Reynolds numbers ranged from 196 to 564 and Womersley (frequency) parameters ranged from 2.00 to 4.1. At the level of the bifurcation, secondary flows were insignificant in the normal plane but strong in the plane of the bifurcation. As a result, two-dimensional velocity fields, reconstructed by vector addition of velocities measured in the plane of the bifurcation, differed markedly from the one-dimensional profiles calculated assuming flow parallel to the vessel axis. In the two-dimensional velocity fields, forward flow was directed toward the flow divider and reversal occurred earliest near the outer wall. Wide spatial and temporal variations in the shear stress at the endothelium are implied by these detailed, in vivo measurements of the bifurcation velocity fields.

Animals↗

Blood velocity profiles in the origin of the canine renal artery and their relevance in the localization and development of atherosclerosis.

Using a 20-MHz 80-channel pulsed Doppler velocimeter and 30-MHz high-resolution echo ultrasound, we investigated the in vivo hemodynamics at the origin of the renal artery by measuring the velocity profiles and bifurcation geometry of a surgically exposed left renal artery in 10 anesthetized dogs. The angle between the aorta and the renal artery ranged from 60 degrees to 90 degrees (mean, 84 degrees) although the bifurcation did not lie in a single anterodorsal plane and the diameter of the renal artery ranged from 1.5 to 3.5 mm (mean, 2.4 mm). Despite different geometries, the velocity profiles in the different aortorenal bifurcations were similar. Although regions of reverse velocity were observed, the net flow in the renal artery was in the forward direction throughout the cardiac cycle. The peak Reynolds' number was 486 +/- 63. The velocity profiles in the proximal renal artery in the plane parallel to the bifurcation showed velocity vectors directed toward the caudal wall throughout the cardiac cycle. Reverse flow, indicating flow separation, was observed near the cranial wall even during systole. When the probe was placed on the cranial wall perpendicular to the wall, a velocity component from the cranial side to the caudal side was observed. At a distance of four diameters from the renal ostia, velocity profiles were almost parabolic. These results indicate that the velocity pattern near the cranial wall at the renal ostia, at which atherosclerotic lesions are prone to develop, are characterized by 1) a low time-averaged shear rate, 2) separation of the flow, and 3) a time-varying oscillation of the flow.

Animals↗

A case-control study of nutrient status and invasive cervical cancer. II. Serologic indicators.

A study of 387 cases and 670 controls from four Latin American countries evaluated the hypothesis that lower serum levels of eight micronutrients were associated with a higher risk of invasive cervical cancer. The serologic analyses were restricted to a sample of subjects with stage I and II disease to minimize effects of the disease on the serologic markers. Ninety-four percent of eligible subjects donated blood samples, which were analyzed for carotenoids, retinol, and tocopherols by high-pressure liquid chromatography. Cases did not differ significantly from controls in mean serum levels of retinol, cryptoxanthin, lycopene, alpha-carotene, lutein, or alpha-tocopherol. The mean level of beta-carotene was lower and the mean level of gamma-tocopherol was higher among cases as compared with controls. After adjustment for age, study site, sexual and reproductive behavior, socioeconomic status, screening practices, detection of human papillomavirus types 16/18, cholesterol, and triglycerides, a trend of decreasing risk was associated with higher levels of beta-carotene (p for trend = 0.05), with the adjusted odds ratio decreasing to 0.72 for the highest versus the lowest quartile. beta-Carotene results were similar by stage of disease, which argues against an effect of disease progression on nutrient values. Unexpectedly, increasing risks were observed as the level of gamma-tocopherol increased (odds ratio = 2.09; p for trend = 0.03); however, levels were higher among stage II cases as compared with stage I cases, suggesting a metabolic alteration resulting from the disease process. The concordance in the strength and direction of the blood and dietary results, presented in the accompanying report (Herrero R, Potischman N, Brinton LA, et al., American Journal of Epidemiology 1991;134:1335-46), supports a role for beta-carotene or foods rich in beta-carotene in the etiology of cervical cancer. This study also indicates that simultaneous analysis using serologic and dietary nutrient indicators allows better discrimination of the association.

Carcinoma, Squamous Cell↗

Hyperinsulinaemia and microvascular angina ("syndrome X")

Glucose and insulin responses to a glucose load in 11 patients with angina attributed to microvascular coronary dysfunction were compared with those in 11 healthy subjects matched for age, sex, and body mass. Stimulated hyperinsulinaemia was demonstrated in the microvascular angina group. The findings suggest a role for increased concentrations of insulin in coronary microvascular dysfunction.

Adult↗

Complementation of DNA repair in xeroderma pigmentosum group A cell extracts by a protein with affinity for damaged DNA.

Complementation group A of xeroderma pigmentosum (XP) represents one of the most prevalent and serious forms of this cancer-prone disorder. Because of a marked defect in DNA excision repair, cells from individuals with XP-A are hypersensitive to the toxic and mutagenic effects of ultraviolet light and many chemical agents. We report here the isolation of the XP-A DNA repair protein by complementation of cell extracts from a repair-defective human XP-A cell line. XP-A protein purified from calf thymus migrates on denaturing gel electrophoresis as a doublet of 40 and 42 kilodaltons. The XP-A protein binds preferentially to ultraviolet light-irradiated DNA, with a preference for damaged over nondamaged nucleotides of approximately 10(3). This strongly suggests that the XP-A protein plays a direct role in the recognition of and incision at lesions in DNA. We further show that this protein corresponds to the product encoded by a recently isolated gene that can restore excision repair to XP-A cells. Thus, excision repair of plasmid DNA by cell extracts sufficiently resembles genomic repair in cells to reveal accurately the repair defect in an inherited disease. The general approach described here can be extended to the identification and isolation of other human DNA repair proteins.

Adult↗

Diet and the risk of in situ cervical cancer among white women in the United States.

A case-control study of women with incident in situ and invasive cervical cancer was conducted during 1982-83 in five US areas reporting to the Comprehensive Cancer Patient Data System: Birmingham, AL; Chicago, IL; Denver, CO; Miami, FL; and Philadelphia, PA. Controls were selected by random-digit dialing and matched to invasive cases on age, race, and telephone exchange. Of the white non-Hispanic in situ cases and controls identified, 229 (78 percent) and 502 (74 percent) were successfully interviewed. Diet was assessed by asking about the usual adult frequency of consumption of 75 food items and the use of vitamin supplements. Included were the major sources of the four micronutrients postulated to reduce the risk of cervical cancer: carotenoids, vitamin A, vitamin C, and folate. Weak inverse associations between risk of in situ disease and intake of carotenoids, vitamin C, folate, fruit, and vegetables/fruits were noted but, with further analysis, these seemed attributable to residual confounding by the multiple lifestyle-related risk factors for this disease and possibly to selection bias. Vitamin A and vegetable intake were unrelated to risk. Dark yellow-orange vegetable consumption and duration of multivitamin use were each strongly related to reduced risk of in situ disease (P for trend = 0.02 and 0.002, respectively) and need to be evaluated in other studies. The absence of persuasive protective effects for the four micronutrients and the similar findings from our analysis of invasive cervical cancer do not concur with other epidemiologic studies and suggest that the role of diet and nutrition in the etiology of cervical cancer is not yet resolved.

Adult↗

Ultrastructure of the normal human placenta.

In this review, the ultrastructure of the normal human chorionic villus is examined and illustrated. The outer covering of trophoblast, comprising the generative cytotrophoblast and the multinucleated syncytiotrophoblast which is derived from it, is described, including such features as the microvillous surface, vesicles and vacuoles, endoplasmic reticulum and secretory droplets. The structure, composition, development and inclusions of the trophoblastic basement membrane are discussed, and the ultrastructure of the various components of the stroma, including reticulum cells, fibroblasts, Hofbauer cells, capillaries and the non-cellular matrix are illustrated and described, with special reference to their inter-relationships and function.

Basement Membrane↗

The bacterial flagellum and flagellar motor: structure, assembly and function.

The bacterial flagellum is a complex multicomponent structure which serves as the propulsive organelle for many species of bacteria. Rotation of the helical flagellar filament, driven by a proton-powered motor embedded in the cell wall, enables the flagellum to function as a screw propeller. It seems likely that almost all of the genes required for flagellar formation and function have been identified. Continuing analysis of the portions of the genome containing these genes may reveal the existence of a few more. Transcription of the flagellar genes is under the control of the products of a single operon, and so these genes constitute a regulon. Other controls, both transcriptional and post-transcriptional, have been identified. Many of these genes have been sequenced, and the information obtained will aid in the design of experiments to clarify the various regulatory mechanisms of the flagellar regulon. The flagellum is composed of several substructures. The long helical filament is connected via the flexible hook to the complex basal body which is located in the cell wall. The filament is composed of many copies of a single protein, and can adopt a number of distinct helical forms. Structural analyses of the filament are adding to our understanding of this dynamic polymer. The component proteins of the hook and filament have all been identified. Continuing studies on the structure of the basal body have revealed the presence of several hitherto unknown basal-body proteins, whose identities and functions have yet to be elucidated. The proteins essential for energizing the motor, the Mot and switch proteins, are thought to exist as multisubunit complexes peripheral to the basal body. These complexes have yet to be identified biochemically or morphologically. Not surprisingly, flagellar assembly is a complex process, occurring in several stages. Assembly occurs in a proximal-to-distal fashion; the basal body is assembled before the hook, and the hook before the filament. This pattern is also maintained within the filament, with monomers added at the distal end of the polymer; the same is presumably true of the other axial components. An exception to this general pattern is assembly of the Mot proteins into the motor, which appears to be possible at any time during flagellar assembly. With the identification of the genes encoding many of the flagellar proteins, the roles of these proteins in assembly is understood, but the function of a number of gene products in flagellar formation remains unknown.(ABSTRACT TRUNCATED AT 400 WORDS)

Bacteria↗

Apoptosis in pancreatic allograft rejection--ultrastructural observations.

Transplantation of pancreaticoduodenal allografts in the PVG.RT1c----PVG.RT1u high responder rat strain combination led to the evolution of haemorrhagic pancreatic necrosis over a rejection course of 8d. A cellular infiltrate consisting mainly of monocytes and macrophages was detectable in the stroma while acinar cells showed degranulation and a progressive decrease in number. From day 4 on apoptotic bodies could be identified ultrastructurally within the exocrine tissue. Apoptosis during pancreas rejection, previously undescribed, contributes to acinar cell loss after transplantation in this model.

Animals↗

Genetic control of the bacterial flagellar regulon.

A number of cis- and trans-acting transcriptional factors in the flagellar regulons of Caulobacter crescentus and Salmonella typhimurium have been identified and characterized to varying degrees over the past year, bringing us closer to understanding the regulations of these complex gene hierarchies.

Caulobacter crescentus↗

Multi-modal selection effects in the study of adult development: a perspective on multivariate, replicated, single-subject, repeated measures designs.

Some of the selection issues that bear upon the conduct of research using multivariate, replicated, single-subject repeated measures designs are examined and their implications for the study of developmental phenomena discussed. The choices of participants, variables, occasions of measurement, etc., made in the conduct of empirical research all involve some kind of selection. These choices, therefore, introduce selection effects into collected data which, in turn, threaten the generalizability of one's conclusions. Data analyzed for single subjects, for example, are suspect regarding generalizability to other individuals. While the limits of generalizability to persons is a concern to be taken seriously, the concern properly applies to all modes of data classification. Discussions of the importance of person selection emphasize representative sampling (of persons) and generalization to populations of persons. Attention to the selection of variables has led to a focus on multivariate approaches to measurement. For the developmentalist, the occasions mode, which is either implicitly or explicitly involved in definitions of change, is especially relevant to concerns about generalizability. Whether one considers stability or change, the effects of temporal selection ought to be a central concern in designing research.

Human Development↗

Origin of stable flies (Diptera: Muscidae) on west Florida beaches: electrophoretic analysis of dispersal.

Stable fly adults were collected from 16 locations within the state of Florida and from locations in six other states in an attempt to determine the source of stable fly populations along Florida beaches on the Gulf of Mexico. Electrophoretic analyses were made of a minimum of 10 enzymes in each of 37 separate populations. Extremely low heterozygosity resulted in an inability to use standard genetic identity and distance procedures for determining the divergence of allopatric populations to establish the source of flies captured from beach areas where reproduction was unlikely. Comparisons of rare alleles in populations grouped geographically, computations of number of possible migrants, and analysis of conditional average frequency of alleles led to the conclusion that flies captured on Florida beaches come from a variety of sites, with locations northwest of the beaches contributing the majority of flies in outbreaks.

Alleles↗

Lectin binding sites in normal, scarred, and lattice dystrophy corneas.

Normal, scarred, and dystrophic corneas were histochemically probed with a panel of 16 lectins by means of an avidin-biotin revealing system. Normal corneal epithelial cells, keratocytes, and endothelial cells expressed at least two distinct N-linked oligosaccharide subsets, of the non-bisected, biantennary and bisected, bi-/triantennary types. Corneal scars stained variably with the lectin subsets described above, and with Maclura pomifera agglutinin. Lattice dystrophy corneas showed a loss of the oligosaccharide expression observed on the plasma membranes of normal epithelial cells, and there was concurrent deposition of extracellular glycoprotein within the corneal stroma, which was of the same oligosaccharide subsets as were lost from the epithelial cell plasma membranes. This extracellular stromal glycoprotein was far more widely deposited than the amyloid and extended well beyond the stromal scarring. We propose that these observations are related and that in lattice corneal dystrophy a glycoprotein(s) is shed from the plasma membranes of epithelial cells and sequestrated within the corneal stroma, where it subsequently stimulates amyloid deposition.

Adult↗