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PubMed1977

A prospective study of angiotensin II pressor responsiveness in pregnancies complicated by chronic essential hypertension.

The present study was designed to (1) outline, in a prospective manner, vascular responsiveness to angiotensin II (A-II) in gravidas with essential hypertension and (2) ascertain if increased sensitivity to infused A-II could be detected before worsening of hypertension developed in those women who were destined to develop superimposed pregnancy-induced hypertension (PIH). Sixty-three women with chronic essential hypertension were studied sequentially throughout pregnancy and two groups of patients were defined on the basis of clinical outcome and serial measurements of vascular reactivity to exogeneously administered A-II. The first group consisted of 29 gravidas with chronic hypertension alone and the second group was composed of 34 patients with chronic hypertension who were destined to develop superimposed PIH. Vascular resistance to infused A-II (more than 7 ng. per kilograms per minute required to increase diastolic blood pressure 20 mm. Hg) was observed from weeks 21 to 27 of gestation in both groups of patients. In the gravidas with essential hypertension alone, resistance to A-II persisted throughout the remainder of pregnancy. However, in those women destined to develop PIH, an increased sensitivity to A-II developed after week 27 of gestation and the differences in mean values between the two groups of patients became highly significant after week 30 of gestation (P less than 0.025). although individual determinations obtained in both groups of patients between weeks 28 and 32 of preganncy suggested that pressor responsiveness to A-II might be used as a screening technique to identify those women destined to develop PIH, additional studies conducted between weeks 28 and 32 of gestation will be required before the efficacy of such a screening technique is clearly established.

Angiotensin II↗

PubMed2004

On the highway measures of driver glance behavior with an example automobile navigation system.

An over-the-road study of visual-manual destination entry using an example original equipment GPS-based navigation system was accomplished in traffic on urban streets and motorways. The evaluation used typical drivers, and a vehicle instrumented to record driver eye glances and fixations, driver control inputs, and lateral lane position. The primary task was to drive in a safe manner, in traffic, while maintaining speed and lateral lane position. As a secondary task, the drivers entered successive destinations while driving, using a touch screen, and at their own pace. They were told there was no need to enter the destination quickly. Results are shown for driver glance behavior, lane keeping performance, and subjective ratings. Overall, the drivers were able to accomplish the destination entry tasks with acceptably short glance durations, acceptable total task times, and with satisfactory subjective ratings for ease of entry.

Automobile Driving↗

PubMed2006

Spinal loading during manual materials handling in a kneeling posture.

Stooped, restricted, kneeling, and other awkward postures adopted during manual materials handling have frequently been associated with LBP onset. However, lift assessment tools have focused on materials handling performed in an upright, or nearly upright standing posture. Unfortunately, many of the tools designed to analyze standing postures are not easily adapted to jobs requiring restricted postures. Therefore, the objective of this study was to evaluate spinal loading during manual materials handing in kneeling postures and determine if those loads can be predicted using simple regression. An EMG-driven biomechanical model, previously validated for upright lifting, was adapted for use in kneeling tasks. Subjects knelt under a 1.07m ceiling and lifted luggage of six weights (6.8, 10.9, 15.0, 19.1, 23.1 and, 27.2kgf) to one of four destination heights (0, 25.4, 53.3, 78.7cm). Spine loading was significantly affected by both destination height and load weight. Destination height increased compression, AP shear and lateral shear by an average of 14.5, 3.7 and 6.6N respectively per cm height increase. Load weight increased compression, AP shear and lateral shear by an average of 83.8, 27.0 and 13.1N respectively per kgf lifted. Regression equations were developed to predict peak spine loading using subject height, load weight and destination height with R(2) values of 0.62, 0.51 and 0.57 for compression, AP and lateral shear respectively.

Adult↗

PubMed2004

Endocrine changes associated with metamorphosis and diapause induction in the yellow-spotted longicorn beetle, Psacothea hilaris.

At 25 degrees C and under a long-day photoperiod, all 5th instar Psacothea hilaris larvae pupate at the next molt. Under a short-day photoperiod, in contrast, they undergo one or two additional larval molts and enter diapause; the 7th instar larvae enter diapause without further molt. The changes in hemolymph juvenile hormone (JH III) titers, JH esterase activity, and ecdysteroid titers in pupation-destined, pre-diapause, and diapause-destined larvae were examined. JH titers of the 5th instar pupation-destined larvae decreased continuously from 1.3 ng/ml and became virtually undetectable on day 13, when JH esterase activity peaked. Ecdysteroids exhibited a small peak on day 8, 1 day before gut purge, and a large peak on day 11, 2 days before the larvae became pre-pupae. The two ecdysteroid peaks are suggested to be associated with pupal commitment and pupation, respectively. JH titers of the 5th instar pre-diapause larvae were maintained at approximately 1.5 ng/ml for 5 days and then increased to form a peak (3.3 ng/ml) on day 11. JH esterase activity remained at a low level throughout. Ecdysteroid levels exhibited a large peak of 40 ng/ml on day 18, coincident with the larval molt to the 6th instar. JH titers of the 7th instar diapause-destined larvae peaked at 1.9 ng/ml on day 3, and a level of approximately 1.1 ng/ml was maintained even 30-60 days into the instar, when they were in diapause. Ecdysteroid titers remained approximately 0.02 ng/ml. Diapause induction in this species was suggested to be a consequence of high JH and low ecdysteroid titers.

Animals↗

PubMed1998

Geographical and diapause-related cold tolerance in the blow fly, Calliphora vicina.

Three geographical strains of the blow fly, Calliphora vicina, were tested for cold tolerance at 0 degrees, -4 degrees and -8 degrees C. Survival to eclosion after 1 to 18 days of cold exposure was greater for diapause-destined larvae than for nondiapause-destined larvae of the two northern strains (Nallikari, Finland 65 degrees N and Edinburgh, Scotland 55 degrees N) but not for the southernmost strain (Barga, Italy 44 degrees N) where no clear differences were apparent. Diapause-destined larvae of the Edinburgh strain were more cold tolerant than those from Nallikari, at both -4 degrees and -8 degrees C, a difference possibly attributable to the long-lasting snow cover in the more northern locality, which might insulate the overwintering soil microclimate. At 0 degrees C, however, Nallikari larvae were more cold tolerant than Edinburgh or Barga. This was also the case for nondiapause-destined larvae, indicating that cold tolerance may occur, in part, independently of the diapause programme. In all three strains diapausing larvae were more cold tolerant than same-age (nondiapausing) pupae. For Nallikari, but not Barga, wandering larvae from short-day exposed flies, therefore initially programmed for diapause, but diverted from the diapause pathway by larval breeding at 19 degrees C, were significantly more cold tolerant than nondiapause larvae from long-day parents, indicating some maternal regulation of larval cold tolerance. There was, however, no evidence for an additional cold hardiness in larvae acclimatised to cold by a gradual reduction of temperature.

Journal Article↗

PubMed2000

HMO penetration and the geographic mobility of practicing physicians.

In this study, we assessed the influence of changes in health maintenance organization (HMO) penetration on the probability that established patient care physicians relocated their practices or left patient care altogether. For physicians who relocated their practices, we also assessed the impact of HMO penetration on their destination choices. We found that larger increases in HMO penetration decreased the probability that medical/surgical specialists in early career stayed in patient care in the same market, but had no impact on generalists, hospital-based specialists, or mid career medical/surgical specialists. We also found that physicians who relocated their practices were much more likely to choose destination markets with the same level of HMO penetration or lower HMO penetration compared with their origin markets than they were to choose destination markets with higher HMO penetration. The largely negligible impact of changes in HMO penetration on established physicians' decisions to relocate their practices or leave patient care is consistent with high relocation and switching costs. Relocating physicians' attraction to destination markets with the same level of HMO penetration as their origin markets suggests that, while physicians' styles of medical practice may adapt to changes in market conditions, learning new practice styles is costly.

Career Mobility↗

PubMed2002

Barriers, facilitators, and access for wheelchair users: substantive and methodologic lessons from a pilot study of environmental effects.

We undertook a month-long intensive pilot study of a sample of adult wheelchair-users in Boston, Massachusetts and Durham, North Carolina, USA. The study had four objectives; to: (1) measure experiences of reaching and failing to reach specific destinations; (2) measure encounters with environmental facilitators and barriers, including both those overcome and not overcome; (3) determine the frequencies of destinations, facilitators, and barriers, and (4) test for consistency between daily reports and retrospective reports. Full participation entailed baseline and exit telephone interviews, and 28 daily telephone contacts. Participants reported reaching a wide range of destinations, most notably, banks, stores and shops, friends' and relatives' homes and health professionals' offices. There was a smaller range of destinations that they could not reach, despite trying; most notably, religious buildings, friends' and relatives' homes and work-places. They encountered an array of barriers, some of which they were able to overcome and others they could not overcome. Reported barriers included personal, interpersonal, and environmental barriers. The 25 subjects completing the study reported a wide range of human, environmental, and technologic support. In general, the consistency among daily, baseline, and exit interviews was high. This study has both substantive and methodologic implications. It suggests that efforts to facilitate social participation by wheelchair-users should focus not only on the built environment, but also on interventions in personal assistance and assistive technology, health promotion and fitness, and programs that improve civility. Methodologically, the data suggest that it is possible to make reliable measures of environmental encounters without the administrative and respondent burden associated with daily interviews.

Activities of Daily Living↗

PubMed2003

Total artificial hearts: bridge to transplantation.

The CardioWest TAH was created and initially tested at the same time as the Thoratec, Novacor, and HeartMate devices. It was designed as a permanent artificial heart and was the first-ever mechanical circulatory device to be used as destination therapy. Twenty years have passed since that early experience. Pneumatic technology is still current and being developed as in existing or new implantable Thoratec VADs the pneumatic HeartMate, and the Abiomed BVS 5000 pumps. Portable pneumatic drivers have been available since 1982, and in recent times have allowed discharge to home of substantial numbers of patients, thus reducing the length of hospital stays and making mechanical device support less expensive to society and more tolerable to patients. Within months, a portable driver for the CardioWest will be available. The documented benefits of the CardioWest TAH include rescue of: critically ill patients with advanced heart failure; patients with biventricular failure especially those with significant right heart failure, elevated pulmonary vascular resistance, or pulmonary edema; patients with renal or hepatic failure secondary to low cardiac output; patients with massive myocardial damage such as those with post-\infarction VSD or irreversible cardiac graft rejection; patients with mechanical valves or native valve disease; and patients with intractable arrhythmias and heart failure. High device outputs with restoration of normal filling pressures result in high perfusion pressures that have led to dramatic recoveries, convalescence, and return to levels of activity compatible with normal life. The average device output with the CardioWest TAH is higher than any other approved or investigational device. The reason for this resides in design simplicity this device has the shortest and largest inflow pathway. Stroke, in the authors' own series, is rare with a linearized rate of 0.068 events per patient year. If the experiences of La Pitie and the University of Arizona are combined, there has been one stroke in 25 patient years (0.04 events/patient year). Serious infections have been rare (12% of patients). No clinical mediastinitis has occurred. Drivelines have healed in tightly and never caused an "ascending" infection. There has not been a case of device endocarditis. Using a broad definition of bleeding, including takeback reoperation for bleeding, bleeding more than 8 units in the first postoperative 24 hours or 5 units over any other 48-hour period, a 25% to 36% incidence has been documented. No cases of fatal exsanguination have resulted, as there have been with the HeartMate. The incidence of bleeding as an adverse event is about 17% lower than the rate reported for the HeartMate VE LVAD, and it is about the same as that reported for Novacor and for Thoratec. Implantation of this device is relatively easy and often done (with attending help) by the authors' residents. If one follows the guidelines for fitting the device, and takes the recommended advice for implantation, hemostasis is excellent and restoration of immediate cardiac function with high flows is nearly automatic. Use of a neopericardium of 0.1 mm EPTFE at the time of implantation assures atraumatic and relatively quick re-entry for transplantation and prevents the normal inflammatory mediastinal reaction that might be desirable in a destination application. In selected patients the CardioWest TAH is the device of choice for bridge to transplantation. When a portable driver becomes available, out of hospital management of CardioWest TAH patients will be feasible and consideration of use of this device for longer term applications, (e.g., "destination therapy,") will be reasonable. A wearable driver, even smaller than a portable, will improve quality of life and expand the patient population that may be therapeutically served with this system. In short, the CardioWest TAH has come nearly full circle. It was first used as a destination device. It has since been used as a bridge to transplantation in nearly 200 patients as the Jarvik-7/Symbion TAH and, since 1993, in over 225 patients as CardioWest. The results have improved with time. Thromboembolism and infection rates have been competitive with currently available devices. Device reliability and durability have been excellent. Survival rates have been very high in a group of perhaps the sickest patients to be supported with any pulsatile device. Pneumatic technology has improved with portability and miniaturization, and there is reason to believe that it will become even better. Application of modern manufacturing techniques to this very simple device raises the possibility of significant manufacturing cost reduction, in an era of prohibitive cost for other devices. All of this establishes the CardioWest as a valuable device for any program that is seriously interested in end-stage heart disease and a likely device for permanent use in appropriately selected patients.

Anticoagulants↗

PubMed1993

Stimulus-response compatability for moving stimuli: perception of affordances or directional coding?

Michaels reported a compatibility effect in which responses were fastest at the destination of a moving stimulus; she interpreted this "destination" compatibility effect in terms of catching actions "afforded" by the stimulus motion. The present study evaluated implications of the catching-affordance account and compared them with those of an account based on spatial coding of relative direction. The destination compatibility effect was obtained when the responses were keypresses rather that catching movements of a joystick and regardless of whether the stimulus expanded, contracted, or only changed location. This effect was a function of relative rather than absolute location of the responses. A similar compatibility effect was obtained when destinations were designated by static arrow stimuli. The results are inconsistent with the catching-affordance account and are best explained by the coding of relative direction.

Adult↗

PubMed2001

GPS computer navigators to shorten EMS response and transport times.

GPS (global positioning satellite system to determine one's position on earth) units have become inexpensive and compact. The purpose of this study is to assess the effectiveness of a GPS enhanced computer street map navigator to improve the ability of EMS drivers in an urban setting to locate their destination and shorten response times. For part I, residential addresses in the city were randomly selected from a telephone directory. Two driver/navigator teams were assigned to drive to the address adhering to speed limits. One team used a standard street map, whereas the other team used a GPS computer navigator. The travel time and distance of the runs were compared. For part II, the computer GPS navigator was placed on an ambulance to supplement their normal methods of navigation to find the address requesting EMS. After the run was completed, EMS providers were interviewed to determine their opinion of whether the GPS navigator was helpful. For part I the results showed that in the 29 initial test runs, comparing the GPS team versus the standard map team, the mean distances traveled were 8.7 versus 9.0 kilometers (not significant) and the mean travel times were 13.5 versus 14.6 minutes (P=.02), respectively. The GPS team arrived faster in 72% runs. For part II the results showed that most EMS providers surveyed noted that the GPS computer navigator enhanced their ability to find the destination and all EMS providers acknowledged that it would enhance their ability to find a destination in an area in which they were unfamiliar. These results suggest that a portable GPS computer navigator system is helpful and can enhance the ability of prehospital care providers to locate their destination. Because these units are accurate and inexpensive, GPS computer navigators may be a valuable tool in reducing pre-hospital transport times.

Ambulances↗

PubMed2000

Directing wayfinders with maps: the effects of gender, age, route complexity, and familiarity with the environment.

The participants were 360 Canadian undergraduates. After receiving written scenarios involving a campus visitor seeking direction to a nearby destination (simple route) or a distant destination (complex route), they drew maps to that destination. The authors varied the visitor's gender, age, and familiarity with campus. They analyzed the content of the students' maps in terms of cardial indicators (compass grid, correct north-south orientation), landmarks, labeled buildings, directional arrows, and supplemental written directions. The men tended to provide more cartographically complete maps than the women, though there were no gender differences in use of landmarks or labeled buildings. The men were significantly more inclined than the women to take visitor characteristics into account, providing more complete maps to visitors navigating complex routes, to newcomers, and to older visitors who were unfamiliar with the campus. The men had more confidence that their maps would successfully lead visitors to their destinations. Route complexity led to greater use of landmarks, labeled buildings, and cardinal indicators. The participants' self-reported familiarity with campus had little effect on their direction giving, except for a greater use of labeled buildings in maps.

Adult↗

PubMed2005

Labor market trajectories and health: a four-year follow-up study of initially fixed-term employees.

With the growth of atypical employment, there is increasing concern about the potential health-damaging effects of unstable employment. This prospective study of Finnish public-sector employees in 1998-2002 examined labor market trajectories and changes in health. At entry, all participants had a fixed-term job contract. Trajectories were measured by exposure to unstable employment during follow-up, destination employment status at the end of follow-up, and the way in which these elements were combined. Nonoptimal self-rated health at baseline was associated with high exposure to unstable employment and unemployment as the destination. After adjustment for health and psychological distress at baseline, a trajectory with stable employment as the destination was associated with a decreased risk of psychological distress at follow-up (odds ratio = 0.68, 95% confidence interval: 0.46, 0.98), whereas a trajectory toward the labor market periphery was related to increased risk of nonoptimal health (odds ratio = 2.54, 95% confidence interval: 1.47, 4.39) when compared with remaining in fixed-term employment. A significant dose-response relation was seen between the measure combining exposure to instability with destination employment status and nonoptimal health. This longitudinal study provides evidence of health-related selection into employment trajectories and suggests that the trajectories themselves carry different health risks.

Adult↗

PubMed2001

Cell migration from the corticostriatal angle to the basal telencephalon in rat embryos.

Lateral cortical stream (LCS) has been described as a flow of cell migration found in the lateral cortex of the embryonic telencephalon of mammals. The destinations of the cell migration were reported as the ventrolateral neocortex, the pyriform cortex, endopyriform nucleus and the claustrum. At the same time, however, other destinations of LCS have been suggested in the ventral telencephalon. Therefore, we investigated the additional destinations of the LCS using a combination of several molecular biological techniques. Using an expression vector of modified green fluorescent protein (GFP) introduced into the ventricular zone (VZ) around the corticostriatal angle, both tangential and radial cell migration was revealed in the LCS. The radial cell migration in the LCS supplied cells to the ventro-lateral neocortex, pyriform cortex and to the level of the lateral olfactory tract. In the second experiment, we injected COS-1 cells transfected with a Sema3A expression vector into one side of the neocortex. The cell supply to the destination of the LCS ceased due to the formation of a large necrosis in the LCS, which was triggered by the Sema3A-COS cell injection, and the dense cell layer in the olfactory tubercle shrunk on the side where COS cells were injected. These data indicated that the majority of neurons in the dense cell layer of the olfactory tubercle reached this point through the LCS. One of the origins of the cells in the LCS would be in the corticostriatal angle.

Animals↗

PubMed2002

Magnetic resonance imaging characterization of hemorrhagic transformation of embolic stroke in the rat.

Intracranial hemorrhage is a critical factor when considering efficacy and safety of thrombolytic intervention after thromboembolic stroke. This study tested whether magnetic resonance imaging could identify tissue for hemorrhagic transformation after the onset of embolic stroke. Rats subjected to embolic stroke with and without recombinant tissue-type plasmogen activator (rt-PA) treatment were followed-up with magnetic resonance imaging using the inverse of the apparent forward transfer rate for magnetization transfer (k(inv)), gadolinium-chelate contrast-enhanced magnetic resonance imaging, and diffusion-, perfusion-, and T2-weighted imaging. Rats with embolic stroke either were treated with rt-PA 1 (n = 16) or 4 hours (n = 13) after stroke onset or were not treated (n = 15). From these groups, at total of 17 rats had intracerebral hemorrhage. Tissue progressing to hemorrhage and adjacent to the site of hemorrhage was analyzed to identify magnetic resonance imaging markers that characterize hemorrhagic transformation. The parameter maps of k(inv) and contrast-enhanced magnetic resonance imaging showed greater sensitivity in the detection of tissue destined for hemorrhagic transformation compared with the apparent diffusion coefficient of water (ADCw) and CBF. In tissue not destined to undergo hemorrhagic transformation, k(inv) maps and contrast-enhanced magnetic resonance imaging exhibited small increases in k(inv) and contrast-enhanced magnetic resonance imaging signal intensity in the area encompassing the territory supplied by the middle cerebral artery. In contrast, large increases in k(inv) and in signal intensity in the contrast-enhanced magnetic resonance images were detected in the region where gross hemorrhage was confirmed histologically. The values of k(inv), T2, and signal intensity in the contrast-enhanced magnetic resonance images were significantly higher in the region destined for hemorrhagic transformation (k(inv), P < or = 0.033 3-24 hours after embolization; T2, P < or = 0.037 24-48 hours; contrast-enhanced magnetic resonance imaging, P < 0.05 4-7 hours) compared with the nonhemorrhagic transformation ischemic region or in the contralateral homologous regions after onset of ischemia. Of these methods, k(inv) shows the most sensitivity in the detection of hemorrhagic transformation soon after embolization. The authors' data suggest that k(inv) and contrast-enhanced magnetic resonance imaging are potentially important methodologies for detecting tissue destined for hemorrhagic transformation.

Animals↗

PubMed1994

Effects of 21-aminosteroid U74389F on skin-flap survival after secondary ischemia.

The effect of 21-aminosteroid, a lazaroid (U74389F), on tissue injury after secondary ischemia was evaluated in 3 x 5 cm island skin flaps in male Sprague-Dawley rats (n = 30). Primary ischemia was produced by arteriovenous occlusion for 1 hour. Eighteen hours later, secondary ischemia was inflicted by 4-hour venous occlusion. The treatment group received intravenous lazaroid (3 mg/kg) 30 minutes before the start of secondary ischemia. The control group received the same volume of citrate buffer vehicle using the same route and schedule. Skin-flap survival was an all-or-none phenomenon and assessed at 7 days after secondary ischemia. Malondialdehyde (MDA) content was measured to determine the occurrence of lipid peroxidation. Myeloperoxidase (MPO) activity was assayed to assess the degree of neutrophil infiltration. Treatment with this lazaroid significantly improved the survival rate from 0 percent (0 of 13) to 53 percent (9 of 17) (p < 0.01). Malondialdehyde content was 62 +/- 10 (mean +/- SEM, n = 4) pmol/mg dry weight in normal skin. Malondialdehyde increased by 3 times normal in the flaps destined to survive and by 13 times in the flaps destined to undergo necrosis (p < 0.001). Myeloperoxidase activity was negligible in normal skin. At the end of secondary ischemia, the flaps destined to survive exhibited a high myeloperoxidase activity. The flaps destined to necrosis showed the enzyme activity 2.2 times more than surviving flaps (p < 0.01). The results suggest that the lazaroid U74389F may improve survival by attenuating neutrophil infiltration and by reducing lipid peroxidation.

Animals↗

PubMed2005

Predicting outcomes after hip fracture repair.

OBJECTIVE: To compare the activities of daily living before and after hip fracture and construct a statistical model for discharge destination and independent walking. The classification accuracy of the model was determined from an independent sample. DESIGN: Prospective study: FIM prefracture, at discharge, and at 6-mo follow-up were obtained from 63 patients who underwent operations for acute hip fractures. A statistical model for discharge destination and independent walking was made and classification accuracy was checked using 78 independent samples. RESULTS: The motor FIM scores at prefracture decreased significantly at discharge (P < 0.0001) and at 6-mo follow-up (P < 0.0001), but at 6-mo follow-up, they had increased significantly compared with those at discharge (P = 0.0103). A mobility subscale was used to predict discharge destination, and mobility and social cognition subscales were related to independent walking. The predictive accuracy was 87%. CONCLUSIONS: Motor FIM scores increase for at least 6 mos after hip fracture, and discharge destination and independent walking were highly predictable from FIM mobility and social cognition subscales.

Activities of Daily Living↗

PubMed2005

Differential gene expression in queen-worker caste determination in bumble-bees.

Investigating how differential gene expression underlies caste determination in the social Hymenoptera is central to understanding how variation in gene expression underlies adaptive phenotypic diversity. We investigated for the first time the association between differential gene expression and queen-worker caste determination in the bumble-bee Bombus terrestris. Using suppression subtractive hybridization we isolated 12 genes that were differentially expressed in queen- and worker-destined larvae. We found that the sets of genes underlying caste differences in larvae and adults failed to overlap greatly. We also found that B. terrestris shares some of the genes whose differential expression is associated with caste determination in the honeybee, Apis mellifera, but their expression patterns were not identical. Instead, we found B. terrestris to exhibit a novel pattern, whereby most genes upregulated (i.e. showing relatively higher levels of expression) in queen-destined larvae early in development were upregulated in worker-destined larvae late in development. Overall, our results suggest that caste determination in B. terrestris involves a difference not so much in the identity of genes expressed by queen- and worker-destined larvae, but primarily in the relative timing of their expression. This conclusion is of potential importance in the further study of phenotypic diversification via differential gene expression.

Animals↗

PubMed2006

Gateway-compatible vectors for plant functional genomics and proteomics.

Gateway cloning technology facilitates high-throughput cloning of target sequences by making use of the bacteriophage lambda site-specific recombination system. Target sequences are first captured in a commercially available "entry vector" and are then recombined into various "destination vectors" for expression in different experimental organisms. Gateway technology has been embraced by a number of plant laboratories that have engineered destination vectors for promoter specificity analyses, protein localization studies, protein/protein interaction studies, constitutive or inducible protein expression studies, gene knockdown by RNA interference, or affinity purification experiments. We review the various types of Gateway destination vectors that are currently available to the plant research community and provide links and references to enable additional information to be obtained concerning these vectors. We also describe a set of "pEarleyGate" plasmid vectors for Agrobacterium-mediated plant transformation that translationally fuse FLAG, HA, cMyc, AcV5 or tandem affinity purification epitope tags onto target proteins, with or without an adjacent fluorescent protein. The oligopeptide epitope tags allow the affinity purification, immunolocalization or immunoprecipitation of recombinant proteins expressed in vivo. We demonstrate the utility of pEarleyGate destination vectors for the expression of epitope-tagged proteins that can be affinity captured or localized by immunofluorescence microscopy. Antibodies detecting the FLAG, HA, cMyc and AcV5 tags show relatively little cross-reaction with endogenous proteins in a variety of monocotyledonous and dicotyledonous plants, suggesting broad utility for the tags and vectors.

Amino Acid Sequence↗