Biomedical subjects

P R Nelson

Publications and source records attributed to P R Nelson.

At least 19 recordsLinked to original sources

PubMed2006

Effect of a flue-curing process that reduces tobacco specific nitrosamines on the tumor promotion in SENCAR mice by cigarette smoke condensate.

A 30-week dermal tumor promotion study was conducted to evaluate the dermal tumor-promoting potential of cigarette smoke condensate (CSC) collected from cigarettes containing flue-cured tobacco cured by a heat-exchange process (HE) relative to that of cigarettes containing flue-cured tobacco cured by the traditional direct-fire process (DF). Heat-exchange process cured tobacco contains significantly lower concentrations of tobacco specific nitrosamines (TSNAs) compared to traditional direct-fire cured tobacco. Mainstream CSCs were collected by cold trap from smoke generators using the Federal Trade Commission puffing regimen. Groups of 40 female SENCAR mice were initiated by a single application of 75 micro g 7,12-dimethylbenz[a]anthracene (DMBA) to the shaved dorsal skin. CSCs were then applied to the skin three times/week for 29 weeks at 9, 18, or 36mg tar/application. End-points included body weights, clinical observations, organ weights, dermal tumor development and histopathology. The numbers of dermal tumors and the numbers of tumor-bearing mice for each CSC were statistically different from the DMBA/acetone control group and increased with increasing dose. When corresponding doses of each CSC were compared, only the DMBA/mid-dose HE CSC group was statistically significantly different (lower) from the corresponding DMBA/mid-dose DF CSC group. In this assay, the dermal tumor-promotion potential of CSC from heat-exchange flue-cured tobacco did not differ from that of traditional direct-fire flue-cured tobacco CSC.

Administration, Cutaneous↗

PubMed2001

Impact of endovascular-assisted in situ saphenous vein bypass technique on hospital costs.

Our initial experience with endovascular-assisted in situ saphenous vein bypass (EISVB) showed patency rates to be comparable to those with conventional in situ bypass, and resulted in a significant reduction in wound-related complications and hospital length of stay (LOS). Here we evaluate the relative costs of these two approaches. Forty-four patients underwent 46 EISVB procedures using endovascular cannulation and coil occlusion of the saphenous vein side branches. Costs for each patient for the operation, the associated hospital stay, and any postoperative care were assessed. These costs were compared to those of the last 46 conventional open in situ bypass procedures as an historical comparison group. The two groups were statistically similar for all parameters except distal outflow target, with the comparison group having statistically more pedal bypasses (p = 0.004). Subset analysis was performed by subdividing each operative group, into those with popliteal and those with distal bypasses. The results of our analysis led us to conclude that the shorter LOS following EISVB more than compensates for the initial cost incurred by the side branch occlusion system. This shorter stay translates into an overall cost savings for EISVB compared to the cost of conventional in situ bypass. The reductions in wound-related morbidity and recovery time postoperatively with EISVB add an additional long-term cost benefit.

Adult↗

PubMed2001

Early results of infragenicular revascularization based solely on duplex arteriography.

PURPOSE: Recent reports have both advocated and questioned the utility of duplex arteriography (DA) as the sole preoperative imaging modality for planning infragenicular revascularization. This study compares the outcome of patients with critical limb ischemia who underwent infragenicular vein grafts on the basis of DA alone versus conventional preoperative contrast arteriography (CA). METHODS: The study group is composed of 23 consecutive patients who underwent infragenicular vein bypass grafting solely on the basis of preoperative DA from 1998 to 1999. They were compared with 50 consecutive patients who underwent infragenicular vein bypass grafting after CA from 1996 to 1998. Peak systolic velocity and end-diastolic velocity of potential target arteries were recorded during DA studies. In situ saphenous vein grafts were used preferentially, and technical adequacy of all grafts was assessed with completion duplex or arteriography. RESULTS: DA and CA groups were comparable on the basis of age and risk factors. In one limb (4%), the target artery selected by DA was abandoned because of dense calcification. No other revision in target or inflow artery was required on the basis of intraoperative completion studies. At 1 year, primary graft patency (78% vs 70%, P =.72) and limb salvage (70% vs 81%, P =.21) were comparable between the two groups. In the DA group, mean preoperative target artery peak systolic velocity in patent versus failed grafts was 49 +/- 18 cm/s versus 31 +/- 9 cm/s (P =.04), whereas mean end-diastolic velocity was 22 +/- 7 cm/s versus 14 +/- 8 cm/s (P =.08). CONCLUSION: Infragenicular revascularization directed by DA alone provides early graft patency and limb salvage rates comparable to similar procedures that are based on CA. Preoperative DA target artery velocities may predict outcome and improve target selection. These initial results justify further clinical testing of DA as the primary imaging modality for planning infragenicular vein grafts.

Angiography↗

PubMed2001

Results of endovascular superficial femoral endarterectomy.

BACKGROUND: Endovascular superficial femoral artery (SFA) endarterectomy with a ring stripper/cutter and distal stenting has been suggested to have a patency comparable with above-knee bypass surgery. We report our initial experience with this technique. METHODS: Seventeen patients (13 men and 4 women; mean age, 64 years) with SFA occlusion and above-knee popliteal reconstitution underwent attempted remote endarterectomy with a ring cutter system combined with primary stenting of the distal end point. Analysis was performed in a prospective manner with patency rates determined by Kaplan-Meier life-table analysis. RESULTS: The indication for operation was claudication in 8 patients, rest pain in 6, and tissue loss in 3. Initial technical success was achieved in 11 patients (65%). Reasons for technical failure included SFA perforation (4), inability to traverse a calcified/diseased segment (1), and inability to retract/remove the ring cutter (1). Life-table analysis of all patients revealed a primary patency at 1 year of 26% +/- 11%. Primary-assisted patency was 38% +/- 12% at 1 year, with 59% of patients ultimately requiring surgical bypass grafting. In patients in whom initial technical success was achieved, the 1-year primary and primary-assisted patency rates were 40% and 59%, respectively. There were four reocclusions requiring surgical revascularization with below-knee popliteal (2) or tibial (2) bypass grafting, 1 symptomatic restenosis requiring repeat angioplasty, and 1 symptomatic restenosis treated conservatively. CONCLUSION: The results of endovascular SFA endarterectomy were disappointing, with technical success in less than two thirds of patients and a 1-year primary patency of only 26%. Remote SFA endarterectomy appears less effective than above-knee femoropopliteal bypass grafting, and after early failure, patients may require more distal revascularization for limb salvage.

Angioplasty↗

PubMed2000

Hand-assisted laparoscopic aortobifemoral bypass grafting.

OBJECTIVE: Aortobifemoral bypass grafting is a durable operation for arterial reconstruction in patients with symptomatic aortoiliac occlusive disease. In several small laparoscopic series technically demanding aortic operations have been described that have not gained widespread acceptance or applicability. To simplify the laparoscopic approach to the aorta, we have developed a technique of aortobifemoral bypass grafting that uses hand-assisted laparoscopic surgery (HALS) to minimize the complexity of aortic dissection and reconstruction. METHODS: Five patients with symptomatic aortoiliac occlusive disease underwent successful HALS aortobifemoral bypass grafting. With the use of a specialized sleeve device (Hand-Port), an operative hand was introduced into the laparoscopic field while pneumoperitoneum was maintained. Laparoscopic dissection of the infrarenal aorta was then performed with retraction provided by the operative hand. Proximal aortic anastomosis was performed with an open technique through the same 7.5-cm Hand-Port incision, and femoral anastomoses were performed in the standard fashion. RESULTS: Five hand-assisted laparoscopic aortobifemoral bypass grafts were performed (two end-to-end, three end-to-side proximal anastomoses). Mean operative time was 231 minutes. Mean blood loss was 440 mL. All patients underwent extubation immediately after surgery, were ambulatory on postoperative day (POD) 1, and were tolerating their diet by POD 3. The mean length of hospital stay was 3.8 days. One patient was discharged on POD 5 and started a clear liquid diet after a self-limiting postoperative ileus. All patients were asymptomatic and back to full activity/work by 14.6 days postoperatively, on average (range, 11-20 days). CONCLUSION: The HALS offers the advantages of tactile feedback, flexible retraction, and the introduction of conventional surgical instruments, all of which extend laparoscopic surgery and its established benefits to a wide array of more complex surgical problems, including major vascular surgery. Ease of performance, shorter hospital stays, and faster recovery times all suggest that HALS may become a valuable adjunct to conventional aortobifemoral bypass grafting.

Adult↗

PubMed2000

Pedal branch artery bypass: a viable limb salvage option.

PURPOSE: We reviewed our experience with pedal branch artery (PBA) bypass to confirm the role of these target arteries for limb salvage and to identify patient and technical factors that may be associated with graft patency and limb salvage. METHODS: In this retrospective study we analyzed 24 vein grafts to PBAs performed from 1988 to 1998 for limb salvage in 23 patients who had no suitable tibial, peroneal, or dorsal pedal target arteries. These PBA grafts were compared with 133 perimalleolar posterior tibial, defined at or below the ankle, or dorsalis pedis bypass grafts performed contemporaneously; the Kaplan-Meier life table was used in the analysis of graft patency and limb salvage. Life table analyses and logistic regression analysis of prognostic patient variables were also performed. RESULTS: The PBA bypass represented 3% of infrainguinal revascularizations for chronic critical limb ischemia at our institution over the study period. Patients who received PBA bypasses were more likely to be male (92% vs. 69%, P =.02) with lower incidences of overt coronary artery disease (33% vs. 50%, P =.12) and stroke (0% vs 15%, P =.04), and a higher incidence of end-stage renal disease (21% vs 8%, P =.06) than those undergoing perimalleolar bypass. Seventeen percent of PBA bypasses were performed with the anterior lateral malleolar artery, a vessel not previously described as a common bypass target. Two-year primary patency and limb salvage for PBA versus perimalleolar bypass was 70% versus 80% (P =.16) and 78% versus 91% (P = .28), respectively. Patency and limb salvage rates were no different in bypasses with above-knee or below-knee inflow arteries. CONCLUSION: An autogenous vein bypass to the PBA, though rarely required, provides acceptable primary patency and limb salvage when compared with perimalleolar tibial artery bypass when no suitable, more proximal target arteries are available. The PBA bypass should be considered before major amputation is undertaken.

Aged↗

PubMed2000

Endovascular in situ bypass decreases morbidity and hospital stay following infrainguinal arterial reconstruction.

PURPOSE: To report the early results of endovascular in situ saphenous vein bypass (EISVB) using side branch coil occlusion. METHODS: Between September 1997 and November 1998, 25 patients (15 men; mean age 70.9 years, range 53-85) with lower limb ischemia were treated with endovascular femorodistal bypass. The saphenous vein was prepared using retrograde valvulotomy and endoscopic cannulation with coil occlusion of the side branches. Duplex graft surveillance was performed at 1, 3, 6, and 12 months. RESULTS: The 25 EISVB procedures consisted of 15 femorodistal popliteal, 7 femorotibial, 2 femoroperoneal, and 1 femorodorsalis pedis in situ saphenous vein reconstructions. Mean operative time was 202 +/- 40 minutes, mean number of side branch coils per case was 5.1 +/- 1.3, and mean number of incisions per case was 2.9 +/- 0.6. Mean hospital length of stay (LOS) was 35 +/- 13 hours (1.4 +/- 0.6 days); 19 (76%) patients were discharged on the first postoperative day. Short-term follow-up (mean 6.2 months, range 2-15) was notable for 2 graft thromboses and 1 graft stenosis; primary and secondary patency rates were 88% and 92%, respectively. Three asymptomatic, persistent arteriovenous fistulas discovered on routine duplex were ligated in the outpatient setting. Only 1 (4%) minor wound complication was encountered. CONCLUSIONS: EISVB provides early patency comparable to conventional in situ infrainguinal bypass. Its distinct advantages, however, are the ability to minimize incision length with resultant reductions in wound-related complications, hospital LOS, and recovery time. EISVB promises to be a useful adjunct in the approach to peripheral vascular insufficiency.

Aged↗

PubMed1999

Use of SHIRPA and discriminant analysis to characterise marked differences in the behavioural phenotype of six inbred mouse strains.

Detailed characterisation of six inbred strains of mice commonly used in transgenic and knockout research was carried out using a battery of behavioural tests (SHIRPA) followed by discriminant analysis of the data. In the primary observation screen, DBA/2 mice were relatively irritable and vocalised during handling. C57BL/6 were hyperactive as measured by transfer arousal, arena activity and touch-escape tests. By contrast, C3H were markedly hypoactive, had significantly enhanced grip strength and were also significantly impaired on the visual placing task. In the elevated plus-maze, BALB/c mice showed the highest level of open arm entries and time spent in the open arms, indicating the lowest level of anxiety. There was a clear dissociation of strains on exploratory activity, as measured in the holeboard test and spontaneous locomotor activity (LMA). DBA/2 mice were hyperactive in LMA but demonstrated relatively low levels of holeboard exploration. None of the six strains learnt the water maze spatial learning task particularly well. C57BL/6 and 129/Sv demonstrated most ability and C3H showed no evidence of having acquired the task. The SHIRPA screening battery and discriminant analysis of the data have enabled us to determine the relevant contribution of a number of behavioural measurements to the marked differences in phenotype of mouse strains. These data confirm the importance of carrying out a comprehensive profile in order to accurately characterise the phenotype of gene-targeted and transgenic mice.

Animals↗

PubMed1999

Adolescents' views regarding sexual history taking.

To address the health needs of adolescents, health care providers need to understand adolescent perceptions of the sexual history taking process. Adolescents (n = 113) were recruited from two sources of health care to complete a questionnaire regarding sexual history taking issues. The results revealed that there were differences in demographics, practice characteristics, and communication strategies between the private office and the hospital clinic. Attitudes and beliefs related to the discussion of sensitive issues were similar. Most adolescents would like health care providers to discuss sensitive topics directly. It is important for health care providers to feel comfortable initiating discussion of sensitive issues directly.

Adolescent↗

PubMed1999

Bacterial counts in canine duodenal fluid after exposure to saline, sodium bicarbonate and hypertonic dextrose solutions used to maintain patency of chronically implanted catheters.

Flushing of intestinal vascular access ports (VAPs) is commonly performed to prevent the problems of blockage and infection, and in this study four different flushing solutions were compared. The growth of bacteria from canine duodenal contents was compared in: 0.9% saline, 50% dextrose, 8.4% sodium bicarbonate (NaHCO3) and 0.01 M phosphate buffered saline (PBS). Duodenal contents from three laboratory beagles were serially diluted in these four solutions, spread plated onto agar at 24 h periods for 7 days and bacterial counts were performed. Immediately after the duodenal juices were added, no significant differences could be seen in bacterial counts with any of the solutions. Over the 7 day period, bacterial numbers greatly increased in saline and phosphate buffered saline, but greatly decreased in dextrose and sodium bicarbonate solutions. Dextrose and sodium bicarbonate appeared to be the most promising flushing solutions tested to minimize infections of associated intestinal VAPs.

Animals↗

PubMed1998

Smooth muscle cell migration and proliferation are mediated by distinct phases of activation of the intracellular messenger mitogen-activated protein kinase.

PURPOSE: Mitogen-activated protein kinase (MAPK) is a ubiquitous signaling protein that has been associated with cellular proliferation; however, its role in cellular migration has not been established. In this study, we investigate the role of MAPK in platelet-derived growth factor (PDGF)-induced migration and proliferation of human vascular smooth muscle cells (SMCs). METHODS: SMC migration was measured using a microchemotaxis assay (4 hours), and proliferation was assessed using 3H-thymidine uptake and cell counts. PD098059 was used as a specific noncompetitive inhibitor of MAPK activation. RESULTS: Coincubation of SMCs with PD098059 resulted in significant inhibition of PDGF-BB (5 ng/ml)-induced SMC chemotaxis and proliferation. The IC50 for both processes was approximately 10 mumol/L with complete inhibition at 50 mumol/L. Stimulation of SMCs with PDGF produced an early peak in MAPK activity followed by a plateau of activity that persisted for 24 hours. We hypothesized that variations in the temporal activation of MAPK might explain the action of this enzyme on these two disparate cellular events. By adding PD098059 at intervals after stimulation of SMCs with PDGF, we demonstrated an association between MAPK activity within the first 15 minutes and SMC migration, whereas MAPK activity between 1 and 4 hours was associated with SMC proliferation. CONCLUSIONS: MAPK activity is essential for both SMC migration and proliferation, and distinct phases of enzyme activation are required to stimulate these two discrete cellular events. Inhibition of this signaling protein may prove to be a useful method for preventing intimal hyperplasia.

Becaplermin↗

PubMed1998

Determination of (+)-alpha-tocopherol in environmental tobacco smoke.

A high-performance liquid chromatographic method is described for the quantitation of (+)-alpha-tocopherol in the particulate phase of environmental tobacco smoke (ETS) collected on a 1-micron pore size Fluoropore membrane. A methanol (MeOH) extract of the membrane, which can be used for four other ETS procedures, is analyzed for (+)-alpha-tocopherol on a reversed-phase column with fluorescence detection at selective wavelengths of 280 nm excitation and 330 nm emission. A mobile phase of MeOH and water is used. The method is reproducible with a relative standard deviation (%) of about 12. Recovery is 88%, and the procedure is capable of detecting greater than 0.04 microgram/m3 (+)-alpha-tocopherol in ETS. A comparison of the ETS from five commercially available cigarettes shows similar (+)-alpha-tocopherol concentrations. A cigarette that primarily heats tobacco yields about 6% of that amount of (+)-alpha-tocopherol found in ETS from tobacco-burning cigarettes. (+)-alpha-Tocopherol can be used as a marker for ETS respirable suspended particles (RSP) because it is found at a consistent amount in ETS RSP of 0.29%. However, sufficient amounts of RSP would have to be generated in order to detect (+)-alpha-tocopherol.

Chromatography, High Pressure Liquid↗

PubMed1998

Comparative studies of the mutagenicity of environmental tobacco smoke from cigarettes that burn or primarily heat tobacco.

The mutagenicity of particulate matter concentrated from environmental tobacco smoke (ETS) from a prototype cigarette that primarily heats tobacco was compared to that of four popular commercially available cigarettes that burn tobacco. ETS was generated by six individuals simultaneously smoking 1 cigarette each in a 20-min time period in a 45 m3 environmental chamber operated in the static mode (without ventilation). Respirable suspended particles (RSP) were collected on polytetrafluoroethylene (PTFE) filters at a flow rate of 3 LPM for 120 min. Less ETS-RSP (86-90%) was emitted by the prototype tobacco-heating cigarette than by the tobacco-burning cigarettes. RSP was extracted from the filters by sequential sonication in acetone and dichloromethane. The acetone extract was dried under nitrogen and the dichloromethane filtrate was added and then dried to obtain ETS-RSP for testing. Mutagenicity was assessed in the microsuspension modification of the Ames Salmonella/microsome assay with strains TA98 and YG1024 in the presence of 5% S9 metabolic activation. The results show that the mutagenic activity of RSP from the prototype cigarette was reduced by 75-83% on a per-mg basis when compared to the commercially available cigarettes and was reduced by 96-98% when calculated as revertants/m3 air under identical smoking conditions.

Air Pollutants↗

PubMed1997

Enhancement of migration by protein kinase Calpha and inhibition of proliferation and cell cycle progression by protein kinase Cdelta in capillary endothelial cells.

Activation of protein kinase C (PKC) induces angiogenesis, migration, and proliferation of endothelial cells (EC), but can also prevent growth factor-induced EC proliferation. To determine whether these disparate effects are mediated by substrates of individual PKC isoenzymes, PKCalpha and PKCdelta were overexpressed in rat microvascular EC. Basal and stimulated migration were enhanced in PKCalpha EC compared with either PKCdelta or control EC. Serum-induced growth of PKCdelta EC was decreased, while that of PKCalpha cells was similar to control EC. Phorbol ester markedly inhibited PKCdelta EC growth but enhanced growth of PKCalpha and control EC. To determine possible causes for this altered proliferation, the effect of PKCdelta on adhesion, mitogen-activated protein kinase activity, and cell cycle progression was measured. Adherence of PKCdelta EC to vitronectin was significantly enhanced. Serum-induced extracellular signal-regulated kinase-2 activity was increased equally in both PKCalpha and PKCdelta EC above that of control, while extracellular signal-regulated kinase-1 activity was similar in all EC. Cell cycle analysis suggested that PKCdelta EC entered S phase inappropriately and were delayed in passage through S phase. Thus, PKCalpha may mediate some proangiogenic effects of PKC activation; conversely, PKCdelta may direct antiangiogenic aspects of overall PKC activation, including slowing of the cell cycle progression.

Animals↗

PubMed1997

Activation of pp60c-src is necessary for human vascular smooth muscle cell migration.

BACKGROUND: The most widely distributed nonreceptor tyrosine kinase is pp60c-src (src), yet the role of this intracellular signaling protein in cell migration has not been defined. Given that smooth muscle cell (SMC) migration is essential for the development of intimal hyperplasia, we investigated the importance of src in locomotion of human vascular SMC. METHODS: SMC migration was evaluated using a microchemotaxis chamber assay and videomicroscopy. Src kinase activity was determined by measuring phosphorylation of a synthetic derivative of p34cdc2, a specific substrate for src. Blocking antibodies to src were introduced using a cytoplasmic microinjection technique. RESULTS: Stimulation of SMC with platelet-derived growth factor (PDGF)-BB and AB resulted in an increase in src activation, whereas PDGF-AA did not consistently enhance src activity. These findings correlated with the ability of the PDGF isotypes to stimulate SMC chemotaxis; PDGF-BB and AB produced 7.4 +/- 0.3- and 5.3 +/- 0.5-fold increases in SMC chemotaxis, whereas PDGF-AA inhibited chemotaxis. SMC migration in response to PDGF-BB and serum was significantly inhibited by intracellular injection of a blocking antibody. CONCLUSIONS: Our findings reveal an association between agonist-induced src activation and chemotaxis. Moreover, an antibody that inhibits src activation dramatically inhibits migration of individual SMC. We conclude that activation of src is necessary for SMC migration. Because of its importance in SMC migration, either molecular or pharmacologic inhibitors of src may be useful in the control of intimal hyperplasia.

Becaplermin↗

PubMed1997

The role of integrins in saphenous vein vascular smooth muscle cell migration.

PURPOSE: Smooth muscle cell (SMC) migration is an essential feature of the intimal hyperplastic process that so frequently limits the patency of vascular reconstructions. The purpose of this investigation was to evaluate the effect of a series of integrins, or cell surface receptors that mediate cellular attachment, on platelet-derived growth factor (PDGF) and extracellular matrix (ECM) protein-induced migration of human SMCs. METHODS: Immunofluorescence staining was used to search for various integrins and subunits on the surface of SMCs derived from human saphenous vein. Chemotaxis and haptotaxis of SMCs to various matrix proteins and PDGF were assayed using a 48-well microchemotaxis chamber in the presence or absence of antibodies that blocked the function of these integrins. RESULTS: Several subunits (beta 1, alpha 2, alpha 5) and one integrin (alpha v beta 3) were identified in saphenous vein SMCs. The beta 1 integrin antibody inhibited chemotaxis to collagen I and IV, laminin, and PDGF. The alpha 2 integrin antibody inhibited collagen I and IV, and laminin-induced chemotaxis. The alpha 5 integrin antibody had no effect on SMC migration. The alpha v beta 3 integrin antibody inhibited chemotaxis to PDGF but not to the ECM proteins. CONCLUSIONS: Integrins are necessary for SMC migration induced by PDGF and ECM proteins. The integrin or subunits responsible for facilitating migration varies with the stimulant. Agonists designed to inhibit integrin function might be used to suppress SMC migration and suppress the formation of intimal hyperplasia.

Cell Movement↗

PubMed1997

Platelet-derived growth factor and extracellular matrix proteins provide a synergistic stimulus for human vascular smooth muscle cell migration.

PURPOSE: Smooth muscle cell (SMC) migration contributes significantly to the hyperplastic response that follows arterial injury. In vitro studies have shown that a number of growth factors and extracellular matrix (ECM) proteins individually stimulate vascular SMC migration. However, after arterial injury, SMCs exist in a complex environment in which they are exposed to many of these proteins simultaneously. The response of SMCs to multiple simultaneous stimuli may differ significantly from their response to any single individual stimulus. In this study, we evaluated the chemotactic response of human vascular SMCs to various combinations of growth factors and ECM proteins. METHODS: Human saphenous vein SMCs were used for all experiments. Using a 4-hour modified Boyden-chamber assay, we evaluated the effect on SMC chemotaxis of combinations of one of three growth factors (platelet-derived growth factor [PDGF]-AB, basic fibroblast growth factor [bFGF], or epidermal growth factor [EGF]), and one of four ECM proteins (fibronectin, laminin, or collagen type I or IV). A standard fluorimetric assay was used to assess changes in intracellular calcium ([Ca2+]i) in response to the various combinations of growth factors and ECM proteins. RESULTS: A simple additive effect was seen between ECM proteins and bFGF or EGF. However, when SMCs were simultaneously exposed to PDGF and ECM proteins, we observed a synergistic increase in chemotaxis. This synergy was evident for all concentrations of collagen type I and IV but only with higher concentrations of fibronectin and laminin. We evaluated whether intracellular calcium may be the signaling pathway through which this synergistic effect is mediated. Although ECM proteins alone did not stimulate a rise in [Ca2+]i, ECM proteins enhanced the early peak in [Ca2+]i induced by PDGF. CONCLUSION: These data show that PDGF acts synergistically with the ECM proteins to promote SMC migration; this effect appears to be specific for PDGF and was not observed with other growth factors. The mechanism responsible for this phenomenon may be a synergistic increase in [Ca2+]i in SMCs simultaneously exposed to both proteins.

Calcium↗

PubMed1996

Role of protein kinase C in attachment, spreading, and migration of human endothelial cells.

Attachment, spreading, and migration of vascular endothelial cells (EC) are necessary for angiogenesis, reendothelialization of an injured artery, or seeding of a prosthetic graft. However, little is known about the signaling pathways that mediate these effects. Protein kinase C (PKC) is a ubiquitous intracellular messenger which we have previously shown to be necessary for EC proliferation (Kent et al., 1995, Circ. Res. 77, 231-238). In this study, we investigate whether activation of PKC is necessary for EC attachment, spreading, and migration. Using human umbilical vein EC, we found that direct activation of PKC with the phorbol ester phorbol 12-myristate-13-acetate enhanced all three processes. Inhibition of PKC by the selective agent, chelerythrine, markedly diminished the ability of EC to attach, spread, and migrate. Depletion of intracellular PKC by downregulation (prolonged exposure of EC to phorbol ester) reduced EC attachment and migration; however, downregulation had no effect on endothelial spreading. PKC is a family of isotypes, each of which may control specific cellular functions. By Western blotting, we identified PKC alpha, beta, delta, epsilon, eta, theta, and zeta isotypes in human EC. Downregulation led to a significant reduction in the quantity of PKC alpha and epsilon. These data demonstrate that activation of PKC is both necessary and sufficient for attachment, spreading, and migration of human EC. An isotype of PKC that is susceptible to downregulation (either alpha and/or epsilon) is at least partially responsible for attachment and migration. Pharmacological activation of PKC may be used as a method to enhance reendothelialization.

Alkaloids↗