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Lesley A Osborn,1 Megan L Brenner,2 Samuel J Prater,1 Laura J Moore3 1Department of Emergency Medicine, The University of Texas Health Science Center, McGovern Medical School, Houston, TX, USA; 2Department of Surgery, The University of California – Riverside, Moreno Valley, CA, USA; 3Department of Surgery, The University of Texas Health Science Center, McGovern Medical School, Houston, TX, USA Abstract: Resuscitative endovascular balloon occlusion of the aorta (REBOA) has recently gained popularity as a minimally invasive alternative to open aortic cross-clamping in the management of patients with non-compressible hemorrhage arising below the diaphragm. The purpose of this review is to provide a description of the technical aspects of REBOA use along with an overview of the current animal and clinical data regarding its use. Keywords: REBOA, hemorrhagic shock, resuscitation

Abstract: Background: A 2021 report of the Aortic Occlusion for Resuscitation in Trauma and Acute Care Surgery multicenter registry described the outcomes of patients treated with Zone 3 resuscitative endovascular balloon occlusion of the aorta (REBOA zone 3). Our study builds upon that report, testing the hypothesis that REBOA zone 3 is associated with better outcomes than REBOA Zone 1 in the immediate treatment of severe, blunt pelvic injuries. Methods: We included adults who underwent aortic occlusion (AO) via REBOA zone 1 or REBOA Zone 3 in the emergency department for severe, blunt pelvic injuries [Abbreviated Injury Score ≥ 3 or pelvic packing/embolization/first 24 hours] in institutions with >10 REBOAs. Adjustment for confounders was accomplished with a Cox proportional hazards model for survival, generalized estimating equations for intensive care unit (ICU)-free days (IFD) and ventilation-free days (VFD) > 0 days, and mixed linear models for continuous outcomes (Glasgow Coma Scale [GCS], Glasgow Outcome Scale [GOS]), accounting for facility clustering. Results: Of 109 eligible patients, 66 (60.6%) underwent REBOA Zone 3 and 43 (39.4%) REBOA Zone 1. There were no differences in demographics, but compared with REBOA Zone 3, REBOA Zone 1 patients were more likely to be admitted to high volume centers and be more severely injured. These patients did not differ in systolic blood pressure (SBP), cardiopulmonary resuscitation in the prehospital/hospital settings, SBP at the start of AO, time to AO start, likelihood of achieving hemodynamic stability or requirement of a second AO. After controlling for confounders, compared with REBOA Zone 3, REBOA Zone 1 was associated with a significantly higher mortality (adjusted hazard ratio, 1.51; 95% confidence interval [CI], 1.04-2.19), but there were no differences in VFD > 0 (adjusted relative risk, 0.66; 95% CI, 0.33-1.31), IFD > 0 (adjusted relative risk, 0.78; 95% CI, 0.39-1.57), discharge GCS (adjusted difference, -1.16; 95% CI, -4.2 to 1.90) or discharge GOS (adjusted difference, -0.67; 95% CI -1.9 to 0.63). Conclusions: This study suggests that compared with REBOA Zone 1, REBOA Zone 3 provides superior survival and is not inferior regarding other adverse outcomes in patients with severe blunt pelvic injuries., (Copyright © 2023 by the Shock Society.)

Resuscitative endovascular balloon occlusion of the aorta (REBOA) is a temporizing maneuver for noncompressible torso hemorrhage. To our knowledge, this single-center brief report provides the most extensive anesthetic data published to date on patients who received REBOA. As anticipated, patients were critically ill, exhibiting lactic acidosis, hypotension, hyperglycemia, hypothermia, and coagulopathy. All patients received blood products during their index operations and received less inhaled anesthetic gas than normally required for healthy patients of the same age. This study serves as an important starting point for clinician education and research into anesthetic management of patients undergoing REBOA.

Background: Resuscitative endovascular balloon occlusion of the aorta (REBOA) is a torso hemorrhage control technique. To expedite deployment, inflation is frequently performed as a blind technique with minimal imaging, which carries a theoretical risk of aortic injury. The objective of this study was to examine the relationship between balloon inflation, deformation and the risk of aortic rupture., Methods: Compliant balloon catheters were incrementally inflated in segments of cadaveric swine aorta. Serial longitudinal and circumferential measurements were recorded, along with the incidence of aortic rupture., Results: Fourteen cadaveric swine aorta segments were tested with mean (±SD) baseline aortic diameter (mm) of 14.2±3.4. Rupture occurred in three aortas. The mean baseline diameters (mm) of the aortic segments that were ruptured were significantly smaller than those that did not rupture (8.9±1.2 vs 15.6±1.9; P<0.001). The maximal circumferential stretch ratios were significantly higher in the aorta segments that ruptured compared with those that did not (1.9±0.1 vs 1.5±0.1; P<0.001). The maximal amount of balloon longitudinal deformation was 80 mm (116% longer than the intended working length)., Conclusions: Inflation of aortic balloon catheters carries an inherent risk of aortic injury, which may be minimized through an understanding of the intrinsic characteristics of the aorta and compliant balloons. Smaller diameter aortic segments undergoing overinflation, particularly beyond a circumferential stretch ratio of 1.8, are at risk of aortic rupture., Level of Evidence: Level II., Competing Interests: Competing interests: JJM and MLB are clinical advisory board members for Prytime Medical Inc.

Introduction: Resuscitative endovascular balloon occlusion of the aorta (REBOA) is a technique that has been shown to provide central vascular control to support proximal aortic pressure and minimize hemorrhage in a wide variety of clinic settings, however the role of REBOA for emergency general surgery is less defined., Case Description: This is a report of a 44 year old man who experienced hemorrhagic shock during video-assisted retroperitoneal debridement (VARD) for necrotizing pancreatitis where REBOA was used to prevent ongoing hemorrhage and death., Discussion: This is the first documented report REBOA being used during pancreatic debridement in the literature and one of the first times it has been used in emergency general surgery. The use of REBOA is an option for those in hemorrhagic shock whom conventional aortic cross-clamping or supra-celiac aortic exposure is either not possible or exceedingly dangerous., Conclusion: REBOA allows for adequate resuscitation and can be used as a bridge to definitive therapy in a range of surgical subspecialties with minimal morbidity and complications. The risks associated with insertion of wires, sheaths, and catheters into the arterial system, as well as the risk of visceral and spinal cord ischemia due to aortic occlusion mandate that the use of this technique be utilized in only appropriate clinical scenarios., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: Blunt iliac arterial injuries (BIAI) require complex management but are rare and poorly studied. We investigated the presentation, management, and outcomes of patients with blunt common or external iliac arterial injuries., Methods: We identified and reviewed 112 patients with BIAI admitted between 2000 and 2011 at a Level I trauma center. Patients with common/external iliac artery injuries (CE group) were primarily analyzed, with patients with injuries of the internal iliac artery or its major branches (IB group) included for comparison of pelvic arterial trauma., Results: Twenty-four patients had CE and 88 had IB injuries. Mean ages (45 ± 19 years) and gender (86% male) were similar between groups. The mean injury severity score was 40 ± 14 (CE, 36 ± 15; IB, 40 ± 14; P = .19), indicating severe trauma. Twenty (83%) of the CE patients presented with signs of leg malperfusion. Admission factors associated with CE injury were crush mechanism of injury (37% vs 17%; P = .03) and pelvic soft tissue trauma (50% vs 15%; P < .01). The CE group had higher early mortality rates, both within 3 hours of admission (50% vs 19%; P = .04) and prior to iliac intervention (42% vs 3%; P < .01). Among those surviving to management, CE patients were more likely to undergo open repair or revascularization (68% vs 3%; P < .01) and had a higher rate of leg amputation (50% vs 6%; P < .01), with 8/12 (67%) culminating in hemipelvectomy. Risk factors for amputation included leg malperfusion, high-grade pelvic fractures, pelvic soft tissue trauma, and increasing leg injury severity. Overall mortality was 40%, and was similar between the injury groups. Among CE patients, need for amputation, pelvic fractures, and wounds were associated with inpatient mortality., Conclusions: This is the largest series to date of blunt CE injuries and demonstrates distinct clinical features and outcomes for these patients. They have high risk for early death and proximal leg amputation. CE injury is specifically associated with serious open pelvic soft tissue injury, which, along with high-grade pelvic fractures, is a risk factor for amputation and death. On-demand emergent endovascular intervention may play an important role in improving management of these complex injuries., (Copyright © 2014 Society for Vascular Surgery. Published by Mosby, Inc. All rights reserved.)

Sucrose synthase is proposed to play an important role in the early stages of tomato fruit (Lycopersicon esculentum Mill.) growth. In this work, the temporal and spatial expression patterns of sucrose synthase during tomato fruit development were investigated. Fruit contained the majority of the sucrose synthase protein and mRNA relative to other organs. Only trace levels of sucrose synthase protein and mRNA were detected in the stem, petiole, and roots. Sucrose synthase mRNA was detected in pistils prior to anthesis, reached peak levels in fruit 5 to 7 d after anthesis (DAA), and was not detectable after 35 DAA. Sucrose synthase protein levels reached a maximum at 20 to 25 DAA and then declined to nondetectable levels after 45 DAA. The lack of coordination between protein and mRNA levels suggests that sucrose synthase expression may be controlled at the levels of both transcription and translation. Sucrose synthase mRNA was differentially localized in the fruit, being most abundant in the mesocarp cells adjacent to the placenta, the columella, and the cells surrounding the vascular bundle. Except around the vascular tissue, the localization of sucrose synthase mRNA positively correlates with starch granule accumulation at the cellular level.

Contrasting evidence has accumulated regarding the role of acid invertase and sucrose synthase in tomato fruit sink establishment and maintenance. In this work the relationships among the activities of sucrose synthase and acid invertase, Lycopersicon esculentum Mill cv UC-82B fruit growth, and starch accumulation were analyzed in fruit at 0 to 39 d after anthesis. Sucrose synthase, but not acid invertase, was found to be positively correlated with tomato fruit relative growth rate and with starch content in the pericarp tissue. A similar association between sucrose synthase activity and starch accumulation was also evident in the basal portion of the stem. Heat-shock treatments, which inhibited the increase in sucrose synthase activity at the beginning of the light period and had no effect on acid invertase activity, were used to examine the importance of sucrose synthase in relation to sucrose metabolism and starch synthesis. After the heat-shock treatment, concomitantly with the suppressed sucrose synthase activity relative to the controls, there was a reduction in sucrose cleavage and starch accumulation. These data substantiate the conclusion that, during the early phases of tomato fruit development, sucrose synthase rather than acid invertase is the dominant enzyme in metabolizing imported sucrose, which in turn plays a part in regulating the import of sucrose into the fruit.

Exogenous applications of gibberellins (GAs) increased the extractable activity of leaf sucrose phosphate synthase (SPS) in soybean (Glycine max [L.]) and spinach (Spinacia oleracea [L.]). The response to GA applications was detectable within 2 h postapplication and was still observed 6 h, 24 h, and 7 d after treatment. When paclobutrazol, a GA biosynthesis inhibitor, was applied to intact soybean and spinach plants, decreased extractable SPS activity resulted within 24 h following the treatment. Different methods of GA application (spray, injection, capillary wick, and excised leaf systems) produced similar effects on SPS activity of soybean leaves. Protein synthesis in soybean leaves appeared to be necessary for GA-promoted SPS activity because gibberellic acid only partially reversed the inhibitory effect of pretreatment with cycloheximide. Levels of SPS protein from crude extracts of spinach plants were measured by a dot blot technique using monoclonal antibodies against SPS. Application of gibberellic acid to spinach leaves increased levels of SPS protein 2 h, 24 h, and 7 d after treatment. The results suggest that, in both soybean and spinach, GA is one of the endogenous hormonal factors that regulate the steady-state level of SPS protein and, hence, its activity.

An important part in the understanding of the regulation of carbon partitioning within the leaf is to investigate the endogenous variations of parameters related to carbon metabolism. This study of diurnal changes in the activities of sucrose-synthesizing enzymes and levels of nonstructural carbohydrates in intact leaves of field-grown soybean plants (Glycine max [L.]) showed pronounced diurnal fluctuations in sucrose phosphate synthase (SPS) activity. However, there was no distinct diurnal change in the activity of fructose-1,6-bisphosphatase (F1,6BPase). SPS activity in leaves from plants grown in controlled environments presented two peaks during the light period. In contrast to field-grown plants, F1,6BPase activity in leaves from growth chamber-grown plants manifested one peak during the first half of the light period. In plants grown under both conditions, sucrose and starch accumulation rates were highest during early hours of the light period. By the end of the dark period, most of the starch was depleted. A pattern of diurnal fluctuations of abscisic acid (ABA) levels in leaves was also observed under all growing conditions. Either imposition of water stress or exogenous applications of ABA inhibited F1,6BPase activity. However, SPS-extractable activity increased following water deficit but did not change in response to ABA treatment. Gibberellin application to intact soybean leaves increased levels of both starch and sucrose. Both gibberellic acid (10(-6)m) and gibberellins 4 and 7 (10(-5)m) increased the activity of SPS but had an inconsistent effect on F1,6BPase. Correlation studies between the activities of SPS and F1,6BPase suggest that these two enzymes are coordinated in their function, but the factors that regulate them may be distinct because they respond differently to certain environmental and physiological changes.

Pea fruit (Pisum sativum L.) is a model system for studying the effect of seeds on fruit growth in order to understand coordination of organ development. The metabolism of (14)C-labeled gibberellin A(12) (GA(12)) by pea pericarp was followed using a method that allows access to the seeds while maintaining pericarp growth in situ. Identification and quantitation of GAs in pea pericarp was accomplished by combined gas chromatography-mass spectrometry following extensive purification of the putative GAs. Here we report for the first time that the metabolism of [(14)C]GA(12) to [(14)C]GA(19) and [(14)C]GA(20) occurs in pericarp of seeded pea fruit. Removal of seeds from the pericarp inhibited the conversion of radiolabeled GA(19) to GA(20) and caused the accumulation of radiolabeled and endogenous GA(19). Deseeded pericarp contained no detectable GA(20), GA(1), or GA(8), whereas pericarp with seeds contained endogenous and radiolabeled GA(20) and endogenous GA(1). These data strongly suggest that seeds are required for normal GA biosynthesis in the pericarp, specifically the conversion of GA(19) to GA(20).

Gibberellins (GAs) are either required for, or at least promote, the growth of the pea (Pisum sativum L.) fruit. Whether the pericarp of the pea fruit produces GAs in situ and/or whether GAs are transported into the pericarp from the developing seeds or maternal plant is currently unknown. The objective of this research was to investigate whether the pericarp tissue contains enzymes capable of metabolizing GAs from [(14)C]GA(12)-7-aldehyde ([(14)C]GA(12)ald) to biologically active GAs. The metabolism of GAs early in the biosynthetic pathway, [(14)C]GA(12) and [(14)C]GA(12)ald, was investigated in pericarp tissue isolated from 4-day-old pea fruits. [(14)C]GA(12)ald was metabolized primarily to [(14)C]GA(12)ald-conjugate, [(14)C]GA(12), [(14)C]GA(53), and polar conjugate-like products by isolated pericarp. In contrast, [(14)C]GA(12) was converted primarily to [(14)C]GA(53) and polar conjugate-like products. Upon further investigations with intact 4-day-old fruits on the plant, [(14)C]GA(12) was found to be converted to a product which copurified with endogenous GA(20). Lastly, [(2)H]GA(20) and [(2)H]GA(1) were recovered 48 hours after application of [(2)H]- and [(14)C]GA(53) to pericarp tissue of intact 3-day-old pea fruits. These results demonstrate that pericarp tissue metabolizes GAs and suggests a function for pericarp GA metabolism during fruit growth.

It has been proposed that abscisic acid (ABA) may stimulate sucrose transport into filling seeds of legumes, potentially regulating seed growth rate. The objective of this study was to determine whether the rate of dry matter accumulation in seeds of soybeans (Glycine max L.) is correlated with the endogenous levels of ABA and sucrose in those sinks. The levels of ABA and sucrose in seed tissues were compared in nine diverse Plant Introduction lines having seed growth rates ranging from 2.5 to 10.0 milligrams dry weight per seed per day. At 14 days after anthesis (DAA), seeds of all genotypes contained less than 2 micrograms of ABA per gram fresh weight. Levels of ABA increased rapidly, however, reaching maxima at 20 to 30 DAA, depending upon tissue type and genotype. ABA accumulated first in seed coats and then in embryos, and ABA maxima were higher in seed coats (8 to 20 micrograms per gram fresh weight) than in embryos (4 to 9 micrograms per gram fresh weight. From 30 to 50 DAA, ABA levels in both tissues decreased to less than 2 micrograms per gram fresh weight. Levels of sucrose were also low early in development, less than 10 milligrams per gram fresh weight at 14 DAA. However, by 30 DAA, sucrose levels in seed coats had increased to 20 milligrams per gram fresh weight and remained fairly constant for the remainder of the filling period. In contrast, sucrose accumulated in embryos throughout the filling period, reaching levels greater than 40 milligrams per gram fresh weight by 50 DAA. Correlation analyses indicated that the level of ABA in seed coats and embryos was not directly correlated to the level of sucrose measured in those tissues or to the rate of seed dry matter accumulation during the linear filling period. Rather, the ubiquitous pattern of ABA accumulation early in development appeared to coincide with water uptake and the rapid expansion of cotyledons occurring at that time. Whole tissue sucrose levels in embryos and seed coats, as well as sucrose levels in the embryo apoplast, were generally not correlated with the rate of dry matter accumulation. Thus, it appears that, in this set of diverse soybean genotypes, seed growth rate was not limited by endogenous concentrations of ABA or sucrose in reproductive tissues.

Mefluidide, N-(2,4-dimethyl-5[([trifluoromethyl]sulfonyl) amino] phenyl)acetamide, a synthetic plant growth regulator, was capable of triggering an increase in endogenous free abscisic acid content when corn (Zea mays L.) plants were grown in a nonstress, day/night, temperature regime (26 degrees C) with sufficient moisture supply. The relevance of such an abscisic acid increase prior to chilling exposure and the water relations during chilling are discussed in reference to the mefluidide protection of the chilled corn plants.

Pea (Pisum sativum L. cv Alsweet) plants were exposed to mixtures of ozone plus sulfur dioxide at different times of the day. Injury, evaluated either as necrosis or chlorophyll, was greatest at midday when stomatal conductance was greatest. Abscisic acid levels were similar over the day, and showed no relation to stomatal conductance.

Cytokinins exported from the root may be involved in the correlative control of plant development. To test this hypothesis in soybean ((Glycine max [L.] Merr. cv. McCall, cv Chippewa 64, and cv Hodgson 78), cytokinins were intercepted en route from the root to the shoot by collecting root pressure exudate from detopped roots. The quantities of four cytokinins in the exudate were studied throughout the development of plants grown in the field and in controlled environment chambers. Zeatin, zeatin riboside, and their dihydro derivatives, dihydrozeatin and dihydrozeatin riboside, were isolated and quantitated using high-performance liquid chromatography.Cytokinin fluxes (pmoles per plant per hour) were independent of exudate flux (grams per plant per hour). All fluxes are averages for a 6- or 8-h collection period. The ribosides accounted for the majority of the observed cytokinin transport. The fluxes of zeatin riboside and dihydrozeatin riboside increased from low levels during vegetative growth to maxima during late flowering or early pod formation. Before the seeds began rapid dry matter accumulation, zeatin riboside and dihydrozeatin riboside fluxes decreased and remained at low levels through maturation. The fluxes of zeatin and dihydrozeatin were low throughout development.No correlation was found between cytokinin fluxes and nodule dry weight or specific nodule activity (acetylene reduction).The timing of distinct peaks in zeatin riboside and dihydrozeatin riboside fluxes during flowering or pod formation suggests that cytokinins exported from the root may function in the regulation of reproductive growth in soybean.

In G2 peas (Pisum sativum L.) apical senescence occurs only in long days (LD), and indeterminate growth is associated with elevated gibberellin (GA) levels in the shoot in short days (SD). Metabolism of GA(12) aldehyde was investigated by feeding shoots grown in SD or LD with [(14)C]GA(12) aldehyde through the cut end of the stem for 0.5 to 6 hours in the light and analyzing the tissue extract by high performance liquid chromatography. More radioactive products were detected than can be accounted for by the two GA metabolic pathways previously known to be present in peas. Three of the major products appear to be GA conjugates, but an additional pathway(s) of GA metabolism may be present. The levels of putative C(20) GAs, [(14)C]GA(53), [(14)C]GA(44), [(14)C]GA(19), and/or [(14)C] GA(17), were all elevated in SD as compared to LD. Putative [(14)C]GA, was slightly higher in LD than in SD. Putative [(14)C]GA(53) was a major metabolite after 30 minutes of treatment in SD but had declined after longer treatment times to be replaced by elevated levels of putative [(14)C] GA(44) and [(14)C]GA(19/17). Metabolism of GA(20) was slow in both photoperiods. Although GA(20) and GA(19) are the major endogenous GAs as determined by gas chromatography-mass spectrometry, putative [(14)C]GA(20) and [(14)C]GA(19) were never major products of [(14)C]GA(12) aldehyde metabolism. Thus, photoperiod acts in G2 peas to change the rate of GA(53) production from GA(12) aldehyde, with the levels of the subsequent GAs on the 13-OH pathway being determined by the amount of GA(53) being produced.

The distribution of abscisic acid (ABA) within maize (Zea mays L.) kernels was studied in kernels from nontreated plants, from plants in which assimilate supply had been altered by source/sink manipulations, and in kernels cultured in vitro on ABA-free media. Prior to growth of the embryo, both the pedicel/placento-chalazal complex and the endosperm contained high concentration of ABA; however, the quantity of ABA in these tissues declined as the concentration in the embryo increased during the early stages of embryo growth. Peaks in the levels of ABA appeared to occur prior to and not concurrent with physiological events during grain filling. During most of the grain filling period, ABA concentration in the embryo was higher than that found in other kernel components. Altering assimilate supply by partial defoliation at two stages of development resulted in variable and transient effects on the relative distribution and concentration of ABA in kernel components. The concentration and distribution of ABA among components of kernels grown in vitro was similar to that observed for field-grown kernels. On the basis of these findings, in situ synthesis of ABA by kernel components is implicated and the putative role of ABA in the regulation of kernel development is discussed.

Photosynthetic rate, ribulose 1,5-bisphosphate carboxylase activity, specific leaf weight, and leaf concentrations of carbohydrates, proteins, chlorophyll, and inorganic phosphate were determined periodically from midbloom until maturity in leaves of soybean plants (Glycine max L., var. Hodgson) from which reproductive and vegetative sinks had been removed 32 hours before measurement, or continuously since midbloom.Leaf photosynthesis, measured in the top of the canopy, was partially inhibited by both sink removal treatments. This inhibition was of constant magnitude from midbloom until maturity.Leaf photosynthesis in the top of the canopy declined from midbloom until maturity in the control as well as in the desinked plants. The decline in photosynthesis was gradual at first, but later became more abrupt. The photosynthetic decline was equally evident in the yellowing leaves of control plants and in the dark green leaves of the continuously desinked plants.Neither the inhibition of photosynthesis by sink removal nor the decline in photosynthetic rate with time was clearly related to any of the measured traits.

Short-term transport studies were conducted using excised whole Zea mays kernels incubated in buffered solutions containing radiolabeled sugars. Following incubation, endosperms were removed and rates of net (14)C-sugar uptake were determined. Endogenous sugar gradients of the kernel were estimated by measuring sugar concentrations in cell sap collected from the pedicel and endosperm. A sugar concentration gradient from the pedicel to the endosperm was found. Uptake rates of (14)C-labeled glucose, fructose, and sucrose were linear over the concentration range of 2 to 200 millimolar. At sugar concentrations greater than 50 millimolar, hexose uptake exceeded sucrose uptake. Metabolic inhibitor studies using carbonylcyanide-m-chlorophenylhydrazone, sodium cyanide, and dinitrophenol and estimates of Q(10) suggest that the transport of sugars into the developing maize endosperm is a passive process. Sucrose was hydrolyzed to glucose and fructose during uptake and in the endosperm was either reconverted to sucrose or incorporated into insoluble matter. These data suggest that the conversion of sucrose to glucose and fructose may play a role in sugar absorption by endosperm. Our data do not indicate that sugars are absorbed actively. Sugar uptake by the endosperm may be regulated by the capacity for sugar utilization (i.e. starch synthesis).

In vitro sugar transport into developing isolated maize embryos was studied. Embryo fresh and dry weight increased concomitantly with endogenous sucrose concentration and glucose uptake throughout development. However, endogenous glucose and fructose concentration and sucrose uptake remained constant. The uptake kinetics of radiolabeled sucrose, glucose, and fructose showed a biphasic dependence on exogenous substrate concentration. Hexose uptake was four to six times greater than sucrose uptake throughout development. Carbonylcyanide-m-chlorophenylhydrazone and dinitrophenol inhibited sucrose and glucose uptake significantly, but 3-O-methyl glucose uptake was less affected. The uptake of 1 millimolar sucrose was strongly pH dependent while glucose was not. Glucose and fructose were readily converted to sucrose and insoluble products soon after absorption into the embryo. Thus, sucrose accumulated, while glucose pools remained low. Based on the findings of this and other studies a model for sugar transport in the developing maize kernel is presented.

Abscisic acid (ABA) was purified from soybean (Glycine max [L.]) seed extract using a preparative high performance liquid chromatography (HPLC) procedure. The preparative procedure was rapid (70 minutes per sample), required no prior partitioning for purification and was quantitative as demonstrated with an internal standard of [2-(14)C]ABA, of which 98.9% was recovered.Following purification by the preparative HPLC procedure, the ABA in a soybean seed extract was quantified using either GLC with an electron capture detector (GLC-EC) or by analytical HPLC with a UV detector. For soybean seed extracts, two analytical HPLC column packing materials were found adequate: muPorasil and muBondapak-NH(2) (Waters Associates). However, with complex tissue extracts, such as soybean leaf and nodule tissues, only GLC-EC had the necessary selectivity and sensitivity.

The level of gibberellin(GA)-like material in cotyledons of soybean (Glycine max L.) was highest at mid-pod fill-about 10 nanograms GA(3) equivalents per gram fresh weight of tissue, assayed in the immersion dwarf rice bioassay. This amount is about 1000-fold less than levels in Pisum and Phaseolus seed, other legume species whose spectrum of endogenous gibberellins (GAs) is well known. The metabolism of [(14)C]-GA(12)-7-aldehyde (GA(12)ald)-the universal GA precursor-by intact, mid-pod-fill, soybean cotyledons and their cell-free extracts was investigated. In 4 hours, extracts converted GA(12)ald to two products-[(14)C]GA(12) (42% yield) and [(14)C]GA(15) (7%). Within 5 minutes, intact embryos converted GA(12)ald to [(14)C]GA(12) and [(14)C]GA(15) in 15% yield; 4 hour incubations afforded at least 22 products (96% total yield). The putative [(14)C]GA(12) was identified as a product of [(14)C]GA(12)ald metabolism on the basis of co-chromatography with authentic GA(12) on a series of reversed and normal phase high pressure liquid chromatography (HPLC) and thin-layer chromatography (TLC) systems, and by a dual feed of the putative [(14)C]GA(12) and authentic [(14)C]GA(12) to cotyledons of both peas and soybeans. The [(14)C]GA(15) was identified as a metabolite of [(14)C]GA(12)ald by capillary gas chromatography (GC)-mass-spectrometry-selected ion monitoring, GC-radiocounting, HPLC, and TLC. By adding the [(14)C] metabolites of [(14)C]GA(12)ald to a different and larger extract (about 0.2 kg fresh weight of soybean reproductive tissue) and purifying endogenous substances co-chromatographing with these metabolites, at least two GA-like substances were obtained and one identified as GA(7) by GC-mass spectrometry. Since [(14)C]GA(9) was not found as a [(14)C]metabolite of [(14)C]GA(12)ald, soybean embryos might have a pathway for biosynthesis of active, C-19 gibberellins like that of the cucurbits; GA(12)ald --> GA(12) --> GA(15) --> GA(24) --> GA(36) --> GA(4) --> GA(7).

Experiments were designed to test the hypothesis that the labeled products recovered from plant tissue incubated with [(14)C]GA(12)-7-aldehyde ([(14)C]GA(12)ald) would serve as appropriate [(14)C]markers for the recovery of naturally-occurring gibberellins (GAs). The [(14)C]GA(12)ald (about 200 millicuries per millimole) was synthesized from pumpkin endosperm using [4,5-(14)C]mevalonic acid. It was added to the adaxial surface of isolated pea cotyledons at 22 days after flowering. Products recovered after 0.5 and 4.0 hour incubations yielded four major peaks which were separated by high performance liquid chromatography (HPLC). These products were purified by multiple-column HPLC using on-line radioactivity detection. They were then added as [(14)C]markers to two unlabeled pea extracts. In general, preparative HPLC followed by further HPLC purification resulted in a single UV-absorbing peak co-eluting with each [(14)C]marker. These [(14)C] and UV-absorbing peaks were shown to contain GA(53), GA(44), GA(20), GA(19), and GA(17) by GC-MS. The finding of GA(53) is novel; all others have previously been found in pea. Endogenous GAs of pea were thus readily detected using [(14)C]GA(12)ald metabolites as [(14)C]markers to recover naturally occurring GAs suggesting that the method may be applicable in detecting naturally occurring GAs in other species.

Injection of [(14)C]tryptophan into one seed in a soybean fruit resulted in recovery of radiolabel in a fraction that cochromatographed with indoleacetic acid (IAA) through three successive high performance liquid chromatography separations. Label was found in the putative IAA fraction in all of the fruit tissues sampled and in the blade of the leaf subtending the pod into which the radiolabeled tryptophan had been injected. This suggested that IAA or an IAA precursor was transported from seeds to other parts of the fruit and to subtending leaves.

Pod removal or petiole girdling, which causes obstruction of translocation, was found in our previous study to cause reduced rates of photosynthesis in soybean leaves due to stomatal closure. The purpose of this research was to determine the involvement of photoassimilate accumulation and leaf abscisic acid (ABA) levels in the mechanism of stomatal closure induced by such treatments.Leaf glucose and sucrose levels increased during the initial 12-hour period after depodding or petiole girdling. Starch, which represents a much larger pool of leaf carbohydrate, was not perceptibly increased above control levels during the 12-hour posttreatment period.When leaflets were exposed to nonphotosynthetic environments (shading or CO(2)-free air) for a 24-hour period after the translocation-obstructing treatments were applied and then returned to normal light or CO(2) concentration, stomatal diffusive conductivity was reduced 65% and 85% with depodding and girdling, respectively. These reductions were comparable to those previously observed without an intervening nonphotosynthetic exposure, thus indicating that photosynthate accumulations were not necessary for the observed response.Free and bound ABA (released on alkaline hydrolysis) were determined by gas liquid chromatography with electron capture detection following preparative high performance liquid chromatography. Free ABA in monitored leaves increased almost 10-fold 48 hours after complete depodding and 25-fold 24 hours after petiole girdling of such leaves. By 3 hours after treatment, in a time course study, free ABA had increased 2-fold above control values in depodded and 5-fold in girdled leaves. Leaf concentrations of bound ABA did not appear to be related to the treatment effects on stomata.Thus, the translocation-obstructing treatments cause an increased level of ABA by a mechanism not involving accumulation of photoassimilate. Increased leaf ABA levels, which were independent of water stress or leaf water potential, appear to be involved in the stomatal closure response. It is suggested that the mechanism of increased leaf ABA levels following translocation-obstruction may be due to an interference with normal translocation of ABA out of leaves.

The fate of exogenously applied, labeled abscisic acid (+/-)-(ABA) was followed in source leaves and taproot sink tissues of sugar beet (Beta vulgaris cv AH-11). The objective was to determine if differential pathways for ABA metabolism exist in source and sink tissues. Tissue discs were incubated for up to 13 hours in a medium containing 1 micromolar labeled ABA. At various time intervals, samples were taken for metabolite determination by reverse-phase high performance liquid chromatography. The labeled metabolites were identified by retention times using an online scintillation counter.Dihydrophaseic acid (DPA) aldopyranoside, DPA, phaseic acid (PA), ABA glucose ester (ABA-GE), and two unidentified compounds were recovered from both tissues. An additional unidentified metabolite was also present in root tissue. Leaf tissue discs exhibited a higher capacity for ABA conjugation, and root discs showed a greater preference for ABA catabolism to PA and DPA. After 4 to 5 hours, ABA incorporation into the various metabolites was proportional to the external ABA concentration in both tissues. But the internal ABA pool size was independent of external concentrations below 10(-6) molar. These results suggested that rates of ABA metabolism was proportional to the rates of uptake in both tissues.

Upon exposure to 2 degrees C day/night (D/N), leaves of Solanum commersonii (Sc) began acclimating on the 4th day from a -5 degrees C (killing temperature) hardy level to -12 degrees C by the 15th day. Leaves of S. tuberosum L. (St) cv ;Red Pontiac' typically failed to acclimate and were always killed at -3 degrees C. Leaves of control (20/15 degrees C, D/N) and treated plants (2 degrees C, D/N) of St showed similar levels of free abscisic acid (ABA) during a 15-day sampling period. In treated Sc plants, however, free ABA contents increased 3-fold on the 4th day and then declined to their initial level thereafter. The increase was not observed in leaves of Sc control plants.Treated St plants showed a slightly higher content of leaf soluble protein than controls. In Sc, leaves of controls maintained relatively constant soluble proteins, but leaves of treated plants showed a distinct increase. This significant increase was initiated on the 4th day, peaked on the 5th day, and remained at a high level throughout the 15-day sampling period.Exogenously applied ABA induced frost hardiness in leaves of Sc plants whether plants were grown under a 20 degrees C or 2 degrees C temperature regime. When cycloheximide was added to the medium of stem-cultured plants at the beginning of 2 degrees C acclimation, or at the beginning of the ABA treatment in the 20 degrees C regime, it completely inhibited the development of frost hardiness. However, when cycloheximide was added to plants on the 5th day during 2 degrees C acclimation, the induction of frost hardiness was not inhibited. The role of ABA in triggering protein synthesis needed to induce frost hardiness is discussed.

Concentrations of abscisic acid (ABA) and indole-3-acetic acid (IAA) in seed parts were determined during reproductive development of soybean plants (Glycine max [L.] Merr. cv ;Chippewa 64'). The concentration of ABA and IAA changed independently in individual seed parts with time. Measurement of the level of ABA and IAA in whole seeds masked the changes which occurred in individual seed tissues. The concentration of ABA was generally highest and that of IAA was generally lowest in the embryonic axis of soybean seeds. In the testa, the IAA concentration was generally highest while the ABA concentration was generally the lowest compared to other parts of the seed.

Water deficits during seed filling decrease seed size in soybean (Glycine max L.). This may result from a reduction in the supply of assimilates from the maternal plant and/or an inhibition of seed metabolism. To determine whether maternal or zygotic factors limited seed growth, we examined the effects of a plant water deficit on the supply of sucrose to and its utilization by developing embryos. Plants were grown in the greenhouse, and water deficits were imposed by withholding water for a period of 6 days during linear seed fill. When water was withheld, leaf water potential decreased rapidly, inhibiting canopy photosynthesis completely within 3 days. However, seed dry weight (nodes 7-11) continued to increase at or near the control rate. The level of total extractable carbohydrates in leaf, stem, and pericarp tissue decreased by 70, 50, and 45%, respectively, indicating that reserves were mobilized to support seed growth. Cotyledon sucrose content decreased from about 60 milligrams per gram dry weight to 30 milligrams per gram dry weight. Similarly, the concentration of sucrose in the interfacial apoplast of the cotyledons decreased from approximately 100 millimolar to 50 millimolar. However, the rate of sucrose accumulation by excised embryos, measured in a short-term in vitro assay, increased in response to the water deficit. These results indicate that both source and sink activity in soybean are altered by water deficits to maintain the flux of assimilates to the developing embryos. This may explain why seed growth is maintained, albeit for a shorter duration, when soybean is exposed to water deficits during the seed filling period.

Pod set in soybean is related to carbon partitioning and may be, at least partially, regulated by abscisic acid (ABA) concentrations. The studies reported here examine the relationship between carbon and ABA partitioning, reproductive abscission and ABA metabolism. The partitioning of radiolabeled ABA and photoassimilates from leaves to flowers and endogenous ABA concentrations were determined in shaded and unshaded reproductive structures. Aluminum foil was gently placed over individual soybean reproductive structures for 48 hours at 0, 4, 12, 17, and 22 days after anthesis (DAA). Shading of flowers at 12, 17, and 22 DAA resulted in significantly reduced concentration of ABA. However, shading had no effect on the catabolism of exogenously supplied [(3)H] ABA. The shading treatment on the first four of the five dates reduced partitioning of photoassimilates and ABA from the subtending leaf to the flower. Shading of reproductive structures also caused a significant reduction in the amount of assimilate exported from the subtending leaf, at 17 DAA. We conclude that shade-induced premature reproductive abscission in soybean is not stimulated by high levels of ABA within reproductive structures, but that ABA may inhibit abscission of reproductive structures by playing a role in preferential assimilate partitioning.

The effect of exogenous abscisic acid (ABA) on the rate of sucrose uptake by soybean (Glycine max L. Merr.) embryos was evaluated in an in vitro system. In addition, the concentrations of endogenous ABA in seeds of three soybean Plant Introduction (PI) lines, differing in seed size, were commpared to their seed growth rates. ABA (10(-7) molar) stimulated in vitro sucrose uptake in soybean (cv ;Clay') embryos removed from plants grown in a controlled environment chamber, but not in embryos removed from field-grown plants of the three PI lines. However, the concentration of ABA in seeds of the three field-grown PI lines correlated well with their in situ seed growth rates and in vitro [(14)C] sucrose uptake rates.Across genotypes, the concentration of ABA in seeds peaked at 8.5 micrograms per gram fresh weight, corresponding to the time of most rapid seed growth rate, and declined to 1.2 micrograms per gram at physiological maturity. Seeds of the large-seeded genotype maintained an ABA concentration at least 50% greater than that of the small-seeded genotype throughout the latter half of seed filling. A higher concentration of ABA was found in seed coats and cotyledons than in embryonic axes. Seed coats of the large-seeded genotype always had a higher concentration of ABA than seed coats of the small-seeded line. It is suggested that this higher concentration of ABA in seed coats of the large-seeded genotype stimulates sucrose unloading into the seed coat apoplast and that ABA in cotyledons may enhance sucrose uptake by the cotyledons.

Indole-3-acetic acid (IAA) promotes an increase in steady-state heat production by corn (Zea mays L.) coleoptile tissue; this increase is associated with an elevation in aerobic respiration rates. A detailed time dependence of the exothermic response to IAA was obtained using flow calorimetry. The latent period and magnitude of response were evaluated as a function of IAA concentration and pH. The data indicate that more than one response may occur. The optimal change in heat production was produced by an IAA concentration of 3·10(-5) M. It was initiated within 5 min after the start of the IAA treatment, and reached a magnitude in excess of 25% of the tissue's basal heat production. Concentrations of IAA greater than 1·10(-4) M resulted in diminished response(s), but the effect was strongly pH dependent. Several possibilities for the increased heat production triggered by IAA are discussed.

A previously unidentified abscisic acid metabolite has been isolated and characterized. (+/-)-[2-(14)C]Abscisic acid was incubated in intact soybean leaves and pods; the radiolabeled metabolite was purified by high performance liquid chromatography with on-line scintillation spectrometry detection. Gas chromatography-mass spectrometry was used to obtain spectra of the acetylated and methyl esterified derivatives. The data were consistent with a proposed dihydrophaseic acid-aldopyranoside identity. Conjugation through the 4'-hydroxyl of dihydrophaseic acid is suggested.

Concentrations of abscisic acid (ABA) and indole-3-acetic acid (IAA) in the second most recently expanded trifoliolate leaf were determined during reproductive development of soybean (Glycine max [L.] Merr cv ;Chippewa 64'). The concentration of ABA in leaves was constant during most of the seed filling period until the seeds began to dry. The concentration of IAA in the leaves decreased throughout development. Removal of pods 36 hours prior to sampling resulted in increased concentrations of ABA in leaves during the period of rapid pod filling but had little effect on the concentration of IAA in leaves. ABA appears to accumulate in leaves after fruit removal only when fruits represent the major sink for photosynthate.ABA and IAA moving acropetally and basipetally in petioles of soybean were estimated using a phloem exudation technique. ABA was found to move mostly in the basipetal direction in petioles (away from laminae). IAA, primarily in the form of ester conjugate(s), was found to be moving acropetally (toward laminae) in petioles. The highest amount of IAA ester(s) was found in petiole exudate during the mid and late stages of seed filling. Removal of fruits 36 hours prior to exudation reduced the amount of IAA ester recovered in exudate, suggesting that fruits were a source of the IAA conjugate in petiole exudate.

The use of immunoaffinity columns containing anti-gibberellin (GA) antibodies for the selective purification of GAs in plant extracts is described. GA(1), GA(3), GA(4), GA(5), GA(7), and GA(9) conjugates to bovine serum albumin were synthesized and used to elicit anti-GA polyclonal antibodies (Abs) in rabbits. Protein A purified rabbit serum, containing a mixture of anti-GA Abs, was immobilized on matrices of Affi-gel 10 or Fast-Flow Sepharose 4B. Columns of these immunosorbents retained a wide range of C-19 GA methyl esters, but no C-20 GA methyl esters. Quantitative recovery of C-19 GA methyl esters was achieved from the columns, which, after reequilibration in buffer, could be reused up to 500 times. The immunosorbents were tested by examination of extracts from immature soybean and pea seeds. GAs were initially purified by passing the extracts through DEAE-cellulose and concentrating them on octadecylsilica. The extracts were methylated and further purified on the mixed anti-GA immunoaffinity columns. GAs were detected and quantified as methyl esters or methyl ester trimethylsilyl ethers by gas chromatography-mass spectrometry-selected ion monitoring. GA(7) was found in soybean seeds, 17 days after anthesis, at low levels (8.8 nanograms per gram fresh weight). C-19 GAs were examined in cotyledons, embryonic axes, and testae of G2 pea seeds harvested 20 days after anthesis. High levels of GA(20) and GA(29) were found in cotyledons (3580 and 310 nanograms per gram fresh weight, respectively) and embryonic axes (5375 and 1430 nanograms per gram) fresh weight, respectively). Lower levels of GA(9) were found in cotyledons and embryonic axes (147 and 161 nanograms per gram fresh weight, respectively). GA(9) was the major GA of testae at levels of 195 nanograms per gram fresh weight. Trace quantities of GA(20) and GA(51) were also observed in testae.

The absorption of Rb(+) by Euglena gracillis var. bacillaris was examined, and an active uptake process could be detected within 1 hour. This was established by experiments that included a time course study and metabolic depression by either reducing the temperature or by addition of 2,4-dinitrophenol. By treating the cells with 0.50 mg/ml chloramphenicol, the rate of glycine incorporation and Rb(+) uptake were inhibited. The possible role of protein synthesis in relation to ion accumulation is discussed.