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Biomedical subjects

W J Hunter

Publications and source records attributed to W J Hunter.

At least 19 recordsLinked to original sources
PubMed2006

A comparison of stent-induced stenosis in coronary and peripheral arteries.

BACKGROUND AND OBJECTIVES: Restenosis is a complication of interventional procedures such as angioplasty and stenting, often limiting the success of these procedures. Knowledge regarding the relative behaviour of different arteries after these procedures is limited, despite the extensive use of different vascular models. Although the results from studies using different vessels are analysed to predict the behaviour of coronary arteries and other vasculature, direct controlled comparisons between different arteries are necessary for a better understanding of the differential response to restenosis. METHODS: This study examines the response to stenting in coronary and internal iliac arteries as characterised by intimal hyperplasia and restenosis. In a swine model of in-stent stenosis, coronary arteries exhibited higher levels of intimal hyperplasia and per cent stenosis than internal iliac arteries. RESULTS: After normalisation for injury score, coronary arteries were found to undergo 47% more intimal hyperplasia (p<0.05), whereas per cent stenosis normalised for injury score tended to be higher (p = 0.01). Other measurements reflecting post-stenting intimal hyperplasia (maximal intimal thickness, medial area) did not exhibit significant differences between the artery groups. CONCLUSIONS: These results show that coronary vessels are more prone to develop significant intimal hyperplasia and subsequent restenosis than internal iliac vessels. A better insight into how different arteries and arterial components behave is important in understanding and developing newer and better therapeutic measures for restenosis.

Animals↗
PubMed2003

Accumulation of nitrite in denitrifying barriers when phosphate is limiting.

Permeable in situ denitrifying barriers can remove nitrate from groundwater. Barriers may be constructed by filling an excavated area with a porous mixture of sand, fine gravel, and substrate or by the injection of a nonaqueous phase substrate into an aquifer. The substrate stimulates the development of a denitrifying microbial community by providing an electron donor. The objective of this study was to determine the ability of denitrifying barriers to function under low-phosphate conditions. Sand columns injected with a soybean oil emulsion were used as laboratory models of denitrifying barriers. When a natural groundwater containing 17 mg l(-1) nitrate-N and 0.009 mg l(-1) phosphate-P was pumped through the columns, only a small amount of nitrate was removed from the water and, in some effluent fractions, 52% to 88% of the influent nitrate had converted to nitrite. Nitrite also accumulated when the phosphate concentration of the groundwater was increased to 0.040 or 0.080 mg l(-1) phosphate-P. Only when a 0.160 mg l(-1) phosphate-P supplement was added to the groundwater was there a loss of nitrate without a large accumulation of nitrite. The addition of solid calcium phosphate or rock phosphate to the sand columns was found to provide adequate phosphate for denitrification in short-term studies. These studies point out the need to ensure that adequate phosphate is present in denitrifying barriers especially when such barriers are used beneath phosphate-binding soils.

Biodegradation, Environmental↗
PubMed2001

Use of vegetable oil in a pilot-scale denitrifying barrier.

Nitrate in drinking water is a hazard to both humans and animals. Contaminated water can cause methemoglobinemia and may pose a cancer risk. Permeable barriers containing innocuous oils, which stimulate denitrification, can remove nitrate from flowing groundwater. For this study, a sand tank (1.1 x 2.0 x 0.085 m in size) containing sand was used as a one-dimensional open-top scale model of an aquifer. A meter-long area near the center of the tank contained sand coated with soybean oil. This region served as a permeable denitrifying barrier. Water containing 20 mg l(-1) nitrate-N was pumped through the barrier at a high flow rate, 1112 l week(-1), for 30 weeks. During the 30-week study, the barrier removed 39% of the total nitrate-N present in the water. The barrier was most efficient during the first 10 weeks of the study when almost all of the nitrate and nitrogen was removed. Efficiency declined with time so that by week 30 almost no nitrate was removed by the system. Nitrite levels in the effluent water remained low throughout the study. Barriers could be used to protect groundwater from nitrate contamination or for the in situ treatment of contaminated water. At the low flow rates that exist in most aquifers, such barriers should be effective at removing nitrate from groundwater for a much longer period of time.

Biodegradation, Environmental↗
PubMed1998

Morphologic and temporal analysis of vascular smooth muscle cell apoptosis induced by c-myc and E1A.

Apoptosis is a physiologic form of cell death present in many disease conditions. When the balance of mitosis versus apoptosis is altered, tumor-like growth or degeneration of tissues may ensue. This appears to occur in several diseases, including those of the cardiovascular system, where apoptosis plays a key role in atherosclerosis and restenosis following angioplasty. Since c-myc is upregulated in the pathogenesis of these diseases, we chose to study the sequential morphologic features of programmed cell death in vascular smooth muscle cells induced by c-myc and by the adenovirus early gene E1A. Morphology and timed events in apoptotic cell cultures were analyzed by scanning electron microscopy, transmission electron microscopy, and time-lapse videomicroscopy. We observed that both c-myc- and E1A-induced apoptosis (in serum-free medium) resulted in numerous, tightly packed clusters of apoptotic blebs, as well as in one or two asymmetrically larger blebs. Transmission electron miscroscopy analysis revealed the larger blebs contained mostly nuclear chromatin, whereas the many smaller fragments often had little or no chromatin. Time-lapse studies showed that apoptosis was induced at a slower rate in cells stably transfected with c-myc versus those stably transfected with E1A. The early changes of apoptosis, including cell shrinkage and intense blebbing, occurred in under 5 min in both cells. Slight alterations such as cell size and further rounding occurred up to 8 h following the initial changes of apoptosis. Rather than being a part of the apoptotic response, release from the culture floor almost entirely resulted from movement of the culture flask. These studies provide a framework of timed morphologic events for future mechanistic investigation into the key aspects of myc- and E1A-induced apoptosis in vascular smooth muscle.

Adenovirus E1A Proteins↗
PubMed1998

Chlamydia pneumoniae and occlusive vascular disease: identification and characterization.

Chlamydia pneumoniae, a respiratory pathogen, has been associated with occlusive vascular disease, including atherosclerosis and intimal hyperplasia, through seroepidemiologic studies. Furthermore, using immunohistochemistry (IHC), polymerase chain reaction (PCR), transmission electron microscopy (TEM), and in situ hybridization, this association has been reconfirmed by detecting this organism in atherosclerotic vascular tissue. This review summarizes and critically analyzes these findings and also discusses various mechanisms of how Chlamydia pneumoniae could be involved in the pathogenesis of occlusive vascular disease. Although more studies are needed to reproduce these results and, possibly, uncover a mechanism, the current literature fails to include detailed methodologies for studying Chlamydia pneumoniae. Therefore, to provide a general standard, we have also outlined specific protocols for IHC, PCR, and TEM. These protocols incorporate essential components from various studies and are presented in a concise and easily adaptable format.

Arterial Occlusive Diseases↗
PubMed1997

The sequence of a symbiotically essential Bradyrhizobium japonicum operon consisting of trpD, trpC and a moaC-like gene.

The 2767 bp BamHI-HindIII fragment specifying the trpDC genes of B. japonicum I-110 was sequenced. The trpD and trpC genes each have three highly conserved 'Crawford' consensus sequences and are part of an operon with three open reading frames (ORFs). The third ORF has a predicted product with 58% amino-acid sequence identity with the gene product of E. coli moaC, a gene encoding an enzyme involved in biosynthesis of the molybdenum cofactor required for the activity of nitrate reductase and other Mo cofactor-requiring enzymes.

Amino Acid Sequence↗
PubMed1997

Functional and morphological evaluation of canine veins following preservation in different storage media.

Injuries of endothelial and smooth muscle cells of autologous vein due to preservation in standard storage media may be responsible for graft failure. The effects of vein preservation with University of Wisconsin solution (UWs) on endothelial and smooth muscle cell function and morphology were compared to the effects of preservation with autologous whole blood (AWB) and normal saline (NS), which are frequently used in cardiovascular surgery. Canine external jugular and common femoral vein segments were preserved in the different solutions at 4 degrees C for 45 min and 24 hr. Rings (4-5 mm in length) from control and preserved veins were evaluated by isometric tension studies at 37 degrees C and by scanning and transmission electron microscopy. Differences between groups were evaluated by Student's t test or Mann-Whitney U test and by analysis of the variance, and considered to be significant at P < 0.05. Sensitivities to norepinephrine (NE) showed that a 45-min vein storage in AWB (5.7 +/- 0.2 mumol/L) but not in NS (5.8 +/- 0.2 mumol/L) or UWs (6.5 +/- 0.2 mumol/L) had a deleterious effect on function of smooth muscle (P < 0.05) when compared to control veins (6.6 +/- 0.2 mumol/L). Maximum contractile responses and sensitivities to NE were significantly altered (P < 0.05) after 24-hr vein storage in AWB (0.09 +/- 0.02 g/mm2 and 5.4 +/- 0.07 mumol/L) and NS (0.12 +/- 0.03 g/mm2 and 5.6 +/- 0.08 mumol/L) but not in UWs (0.36 +/- 0.06 g/mm2 and 6.4 +/- 0.07 mumol/L). With both storage times, acetylcholine-induced endothelium-dependent maximum relaxations and sensitivities were significantly reduced (P < 0.05) in veins stored in AWB and NS, but not in UWs, compared with controls. Similarly, transmission electron microscopy revealed marked neutrophil migration beneath the intimal surface of vessels and extensive separation and desquamation of endothelial cells with exposure of subendothelial structures in veins stored in AWB and NS. The results suggest that UWs is a suitable storage medium when compared to AWB and NS.

Animals↗
PubMed1997

Short-term preservation of autogenous vein grafts: effectiveness of University of Wisconsin solution.

BACKGROUND: Suboptimal preservation of autologous veins in storage solutions causes endothelial cell damage that can contribute to graft failure. The purpose of this study was to compare the effects of short-term storage of veins in autologous whole blood (AWB), 0.9% normal saline solution (NS), and University of Wisconsin solution (UWs) on vein structure and function after grafting. METHODS: Autogenous jugular and femoral veins were atraumatically harvested from mongrel dogs. One vein segment was immediately implanted to serve as a control, and the other segments were stored for 45 minutes in AWB, NS, or UWs. The veins were implanted as reversed interposition graft in the carotid or femoral arteries. After 6 weeks light and scanning electron microscopy and isometric tension studies were performed on explanted vein grafts. RESULTS: Morphologic studies revealed an intact endothelium that stained positively for factor VIII. Intimal thickness was similar between controls (48 +/- 12 microns) and veins stored in UWs (53 +/- 8 microns) (p = not significant), but it was significantly increased in veins stored in AWB (151 +/- 29 microns) and NS (149 +/- 18 microns) (p < 0.05). Sensitivity and maximum contraction to norepinephrine were not altered in veins preserved in UWs (6.0 +/- 0.1 mumol/L and 0.19 +/- 0.02 gm/mm2) but were significantly reduced (p < 0.05) in those stored in AWB (7.2 +/- 0.1 mumol/L and 0.08 +/- 0.02 gm/mm2) and NS (7.0 +/- 0.3 mumol/L and 0.09 +/- 0.02 gm/mm2) compared with controls (5.9 +/- 0.2 mumol/L and 0.20 +/- 0.02 gm/mm2). The sensitivity and maximum relaxation to acetylcholine and sodium nitroprusside of veins preserved in AWB, NS, and UWs were similar to controls (p = not significant). CONCLUSIONS: Vein storage in UWs preserves smooth muscle cell function compared with veins stored in NS or AWB. Therefore UWs is a more suitable medium for short-term preservation of veins in cardiovascular operation.

Adenosine↗
PubMed1997

Morphological and biochemical characterization and analysis of apoptosis.

Apoptosis is a form of programmed cell death serving physiologic and homeostatic functions. However, recent evidence implicating apoptosis in the etiology and pathophysiology of known human diseases, such as heart diseases, cancer, AIDS, and neurodegenerative and autoimmune diseases are continually surfacing. This has spawned the need for identifying which methods are the most effective and well accepted to decipher its presence in a variety of research settings. We have therefore detailed the morphology and biochemical features of apoptotic cell death, with an emphasis on discriminating it from necrosis. In addition, we describe specific and selective techniques which are optimal to target hallmark apoptotic features, such as microscopy, Annexin V labeling, in situ nick-end labeling (TUNEL), and DNA fragmentation analysis by gel electrophoresis and ELISA for oligonucleosome-sized DNA. The advantages and disadvantages of each technique are discussed, as well as their experimental importance relative to one another. The methods have been described in a stepwise fashion, and can readily be applied in the majority of cell systems. Whether working on the tissue or single cell level, these methods are highly effective in qualifying and quantifying apoptosis. The application of these methods in conjunction with molecular techniques can further delineate the underlying mechanisms of apoptosis.

Acquired Immunodeficiency Syndrome↗
PubMed1996

Collagen COL4A3 knockout: a mouse model for autosomal Alport syndrome.

A mouse model for the autosomal form of Alport syndrome was produced. These mice develop a progressive glomerulonephritis with microhematuria and proteinuria, consistent with the human disease. End-stage renal disease develops at approximately 14 weeks of age. TEM analysis of the glomerular basement membranes (GBM) during development of renal pathology revealed focal multilaminated thickening and thinning beginning in the external capillary loops at 4 weeks and spreading throughout the GBM by 8 weeks. By 14 weeks, half of the glomeruli were fibrotic with collapsed capillaries. Immunofluorescence analysis of the GBM showed the absence of type IV collagen alpha-3, alpha-4, and alpha-5 chains and a persistence of alpha-1 and alpha-2 chains (these chains normally localize to the mesangial matrix). Northern blot analysis using probes specific for the collagen chains illustrate the absence of COL4A3 in the knockout, whereas mRNAs for the remaining chains are unchanged. An accumulation of fibronectin, heparan sulfate proteoglycan, laminin-1, and entactin was observed in the GBM of the affected animals. The temporal and spatial pattern of accumulation was consistent with that for thickening of the GBM as observed by TEM. Thus, expression of these basement membrane-associated proteins may be involved in the progression of Alport renal disease pathogenesis. The levels of mRNAs encoding the basement membrane-associated proteins at 7 weeks were unchanged.

Animals↗
PubMed1996

Malpractice protection: communication of diagnostic uncertainty.

Malpractice claims against pathologists for misdiagnosis have been sharply rising, especially in the areas of breast fine-needle aspirations (BFNAs) and cervical (Pap) smears. The current state of medical malpractice law is reviewed as it relates to pathologists' anatomic reports. Communication is one of the best medical malpractice prevention tools. This article examines anatomic pathology reports in terms of the merits of communicating diagnostic error rates to the clinician/patient. In the areas of BFNAs and cervical smears, dissemination of diagnostic error rates in the cytology report is recommended. This would help safeguard against malpractice liability being imposed without showing a deviation by the cytopathologist from reasonable practice standards.

Biopsy, Needle↗
PubMed1996

Fresh vein allograft survival in dogs after cyclosporine treatment.

Synthetic grafts are widely used for peripheral arterial reconstructions when autologous veins are not available, but their results have not been satisfactory. Venous allograft may be used as an alternative to synthetic prostheses. The aim of the study was to explore the immunosuppressive efficacy of Cyclosporine A (CyA) as a means of preventing venous allograft failures and rejection. We utilized 56 mongrel dogs. Immunological incompatibility was checked with the skin graft method. Donor inferior vena cava was transplanted into the infrarenal abdominal aorta of recipient animals. One group (group 1, 10 dogs) served as a control and three groups received CyA treatment regimens. Group 2 (10 dogs) received postoperative oral CyA treatment for 30 days. Group 3 (12 dogs) received a vein graft pretreated with a CyA solution without postoperative immunosuppressive therapy. Group 4 (9 dogs) received a vein graft pretreated with a CyA solution and postoperative CyA treatment for 30 days. Allografts were examined at 30 days for patency, aneurysmal dilatation, gross structural changes, inflammatory response, and lymphocytic infiltration. Sex chromatine assessment determined the origin (donor or recipient) of the endothelial cells. The allografts from groups 1 and 3 showed significant aneurysmal dilatation and perivenous inflammation when compared to dogs treated with oral CyA therapy (P < 0.0002). Moreover allografts treated with CyA therapy had a better-developed venous neointima (P < 0.009) with less fibrin (P < 0.02) and thinner medial (P < 0.0009) with less fibrin (P < 0.02), and thinner medial (P < 0.0009) and adventitial layers (P < 0.02). No significant differences were observed in neointimal thickness among the four groups. Lymphocytic infiltration was greater in the group of animals who did not receive oral CyA therapy (P < 0.0004). Barr bodies status showed significant differences between oral CyA treated groups and nontreated groups (P < 0.0003). Oral CyA therapy reduced aneurysmal dilatation and immunological response, promoted the development of a neoendothelium, and preserved the structure of the venous layers. Graft pretreatment with CyA flushing did not have a significant immunosuppressive effect.

Animals↗
PubMed1996

A noninjury, diet-induced swine model of atherosclerosis for cardiovascular-interventional research.

To investigate whether atherosclerotic vascular disease in the microswine model can be induced by atherogenic diet alone and does not require balloon injury or endothelial denudation as widely stated in the literature, 28 female Yucatan microswine were fed a high-fat, high-cholesterol diet, including 2% sodium cholate, for an average of 310 +/- 13 days. Four control swine were placed on a regular diet for an average of 287.2 +/- 7.8 days. Selective coronary arteriography and morphologic and histologic studies were performed at the end of this period. Coronary arteries were fixed in vivo by pressure perfusion of formalin. Angiograms and sequential histologic sections were reviewed by a double-blind team. The angiography did not show apparent disease in all vessels but generally revealed mild irregularity. Quantitatively, there was a 30.5 +/- 3.5% stenosis (mean +/- standard error, P < 0.05 vs. control) in left anterior descending (LAD), 40.7 +/- 4.5% of stenosis in right coronary artery (RCA) (P < 0.01 vs. control), and 24.8 +/- 3.7% of stenosis in left circumflex artery (LCX). The lesions were eccentric in 95% of LCA, 95.8% of RCA, and 75% of LCX, and the remainder were concentric lesions. Typical lesions were characterized by significant intimal proliferation, cholesterol clefts, necrotic cores, heavy extracellular fat deposition, and calcification. Control animals had only occasional, minimal intimal lipid deposition in coronary arteries. These findings suggest that the Yucatan microswine is an ideal coronary atherosclerosis animal model for vascular research. Lesions can be induced by atherogenic diet alone. Cholesterol uptake is increased by adding sodium cholate to the feed. Moreover, balloon injury of the intima or media is not required to induce significant atherosclerotic lesions in coronary arteries.

Animals↗
PubMed1995

University of Wisconsin solution effects on intimal proliferation in canine autogenous vein grafts.

This study was carried out to investigate if the University of Wisconsin solution (UWs) is suitable for long-term preservation of autogenous vein grafts (AVG) prior to transplantation, compared to autologous whole blood (AWB) and normal saline (NS). Autogenous jugular and femoral veins were harvested from adult mongrel dogs, using a "no-touch" technique. One segment of vein was immediately implanted to serve as a control, while other segments were stored for 24 hr at 4 degrees C in AWB, NS, or UWs. The control and the preserved veins were implanted as reversed interposition grafts in the common carotid or femoral artery positions. After 6 weeks, scanning electron microscopy revealed an intact endothelial cell monolayer in all vein grafts that was stained positively for factor VIII. The degree of intimal thickening, as assessed by light microscopy in the middle position of the grafts, was similar in controls (52.0 +/- 15.8 microns) and in veins stored in UWs (58.5 +/- 16.1 microns), but it was significantly increased (P < 0.05) in veins preserved in NS (198.9 +/- 19.5 microns) and in AWB (312.0 +/- 171.6 microns). Isometric tension studies revealed that maximum contraction and sensitivities (assessed by EC50) to norepinephrine were significantly reduced (P < 0.05) in AVG stored in AWB (0.05 +/- 0.02 g/mm2 and 5.5 +/- 2.8 microM), but not in UWs (0.16 +/- 0.03 g/mm2 0.92 +/- 0.34 microM) and NS (0.09 +/- 0.03 g/mm2 and 3.0 +/- 1.1 microM), compared with controls (0.17 +/- 0.03 g/mm2 and 0.99 +/- 0.38 microM). Acetylcholine (Ach)-induced maximum relaxations were similar in all of the veins.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1994

Penicillin allergies.

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Adverse Drug Reaction Reporting Systems↗