Deceased donor uterus retrieval: A novel technique and workflow.

Uterus transplantation has proven successful when performed with a living donor. Subsequently, interest in the novel field of reproductive transplantation is growing. The procedure is still considered experimental, with fewer than 25 cases performed worldwide, and the techniques of both uterus procurement and transplantation are still developing. We detail a new approach to deceased donor uterus procurement. In contrast to reported techniques and our own initial experience, in which the deceased donor uterus was procured post cross-clamp and after other organs were procured, our approach now is to perform the uterus procurement prior to the procurement of other organs in a multiorgan donor and hence prior to cross-clamp. We describe our practical experience in developing and implementing the logistical workflow for deceased donor uterus procurement in a deceased multiorgan donor setting.

Characterization of BioID tagging systems in budding yeast and exploring the interactome of the Ccr4-Not complex.

The modified E. coli biotin ligase BirA* was the first developed for proximity labeling of proteins (BioID). However, it has low activity at temperatures below 37°C, which reduces its effectiveness in organisms growing at lower temperatures, such as budding yeast. Multiple derivatives of the enzymes have been engineered, but a comparison of these variations of biotin ligases has not been reported in Saccharomyces cerevisiae. Here, we designed a suite of vectors to compare the activities of biotin ligase enzymes in yeast. We found that the newer TurboID versions were the most effective at labeling proteins, but they displayed low constitutive activity from biotin contained in the culture medium. We describe a simple strategy to express free BioID enzymes in cells that can be used as an appropriate control in BioID studies to account for the promiscuous labeling of proteins caused by random interactions between bait-BioID enzymes in cells. We also describe chemically-induced BioID systems exploiting the rapamycin-stabilized FRB-FKBP interaction. Finally, we used the TurboID version of the enzyme to explore the interactome of different subunits of the Ccr4-Not gene regulatory complex. We find that Ccr4-Not predominantly labeled cytoplasmic mRNA regulators, consistent with its function in mRNA decay and translation quality control in this cell compartment.

Evaluation of late vegetative and reproductive stage soybeans for resistance to soybean aphid (Hemiptera: Aphididae).

The soybean aphid, Aphis glycines Matsumura, has become the most significant soybean [Glycine max (L.) Merrill] insect pest in the north central soybean production region of North America. The objectives of this research were to measure selected genotypes for resistance to the soybean aphid in the later vegetative and reproductive stages under field conditions, and confirm the presence of tolerance in KS4202. The results from 2007 to 2011 indicate that KS4202 can support aphid populations with minimal yield loss at levels where significant yield loss would be expected in most other genotypes. The common Nebraska cultivar, 'Asgrow 2703', appears to show signs of tolerance as well. None of the yield parameters were significantly different between the aphid infested and noninfested treatments. Based on our results, genotypes may compensate for aphid feeding in different ways. Asgrow 2703 appears to produce a similar number of seeds as its noninfested counterpart, although the seeds produced are slightly smaller. Field evaluation of tolerance in KS4202 indicated a yield loss of only 13% at 34,585-53,508 cumulative aphid-days, when 24-36% yield loss would have been expected.

Effects of thiamethoxam seed treatments on soybean aphid (Hemiptera: Aphididae) feeding behavior.

Since its discovery in North America in 2000, the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), has rapidly become an important pest of soybean [Glycine max (L.) Merrill], sometimes resulting in significant yield losses. Previous research has documented the toxicity of neonicotinoid seed treatments to soybean aphids, but control under field conditions has been inconsistent. Imidacloprid, a popular neonicotinoid insecticide, has been shown to exhibit antifeedant effects on aphids. Antifeedant activity has not been demonstrated for other neonicotinoids, including thiamethoxam. This research investigated the effects of a thiamethoxam seed treatment on soybean aphid feeding behavior by using electronic penetration graphs (EPG) to visualize stylet penetration behavior. Soybean aphid feeding behavior was assessed for 9 h on thiamethoxam-treated and untreated soybeans (V2 and V4 stages). Because results were inconclusive from initial experiments, a study was conducted to document the effects of thiamethoxam-treated soybeans on soybean aphid survival. The seed treatment was shown to negatively affect aphid survival at 4, 8, and 11 d after aphid introduction. A subsequent EPG study then was designed to document soybean aphid feeding behavior for 15 h, after an initial exposure of 9 h to thiamethoxam-treated soybeans. In this study, the exposed aphids exhibited significant differences in feeding behavior compared with those aphids feeding on untreated soybeans. Soybean aphids on thiamethoxam-treated soybeans spent significantly less time feeding in the sieve element phase, with a greater duration of nonprobing events. These studies suggest soybean aphids are unable to ingest phloem sap, which may be another important element in seed treatment protection.

Categorizing the resistance of soybean genotypes to the soybean aphid (Hemiptera: Aphididae).

We evaluated selected soybean, Glycine max (L.) Merr., genotypes during their reproductive stages for resistance to the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), under greenhouse conditions and documented the categories of aphid-resistant soybean. Two screening studies were performed to assess the level of resistance to the soybean aphid on six soybean genotypes during the reproductive stages of development. Significant differences in aphid damage ratings were detected among the soybean evaluated in the screening studies. Three genotypes (KS4202, K-1639-2, and K1621) were considered moderately resistant based on the assessed damage ratings. Two of these genotypes (K-1639-2 and KS4202), along with a commercial variety ('Asgrow 2703') were used in a follow-up greenhouse study to test for antibiosis and tolerance. For the antibiosis evaluation, KS4202 had significantly more nymphs than Asgrow 2703 and K-1639-2. In fact, KS4202 had a threefold difference in the number of nymphs compared with Asgrow 2703 (81.8 +/- 14.7 and 26.2 +/- 13.9 nymphs, respectively) and a fivefold difference compared with K-1639-2 (15.6 +/- 13.9). Although not significant, Asgrow 2703 had more nymphs than K-1639-2. The lower aphid numbers on infested K-1639-2 plants compared with aphid numbers on Asgrow 2703 and KS4202 plants indicates antibiosis for this genotype. No significant differences in average seed weight, number of seeds per pod, or plant damage were observed between infested and control KS4202 plants; however, significant differences in biomass, total seed weight, number of pods per plant, and number of seeds per plant were detected.

Examination of the DNA-binding ability of estrogen receptor in whole cells: implications for hormone-independent transactivation and the actions of antiestrogens.

We describe an assay employing the competitive binding of estrogen receptor (ER) with basal transcription factors on a constitutive promoter (cytomegalovirus-hormone response element[s]-chloramphenicol acetyltransferase [CMV-(HRE)n-CAT, containing a hormone response element(s) between the TATA box and the start site of transcription]) to examine the DNA-binding ability of the human ER in whole cells. We used this promoter interference assay to examine the DNA binding of ER in cell lines containing high and low levels of endogenous ER, as well as in CHO cells expressing wild-type and mutant ERs from cotransfected expression vectors. The ER is capable of binding to the promoter interference constructs in the absence of added ligand, and estrogen (estradiol) or antiestrogen (trans-hydroxytamoxifen or ICI 164,384) enhances or stabilizes this interaction. The binding of unoccupied ER to reporter gene activation plasmids results in ligand-independent transactivation, presumably due to the TAF-1 function of the receptor. DNA binding of ER in the absence of ligand is observed in cells containing endogenous ER, or expressed ER, and occurs in cells with high or low receptor contents. Although estrogen- and antiestrogen-occupied ER complexes bind to DNA and reduce the template promoter activity, the extent of suppression achieved by ICI-bound ERs is consistently less than that achieved with the other ligands, presumably caused by the fact that ICI rapidly reduces the level of ER in most of the cells examined. However, the ICI-ER complexes that remain are in sufficient quantity to bind to gene activation reporter constructs, and in these cells, ICI still behaves as a pure antagonist of gene transcription and does not activate reporter genes. Hence, obstruction of ER DNA binding or reduction of ER in target cells may contribute to, but cannot fully explain, the pure antagonist character of the antiestrogen ICI 164,384. In addition, DNA binding by the ER alone is clearly not sufficient for ensuring full activation of transcription and argues for an intermediate in the receptor activation of promoters.

Analysis of TAF90 mutants displaying allele-specific and broad defects in transcription.

Yeast TAF90p is a component of at least two transcription regulatory complexes, the general transcription factor TFIID and the Spt-Ada-Gcn5 histone acetyltransferase complex (SAGA). Broad transcription defects have been observed in mutants of other TAF(II)s shared by TFIID and SAGA but not in the only two TAF90 mutants isolated to date. Given that the numbers of mutants analyzed thus far are small, we isolated and characterized 11 temperature-sensitive mutants of TAF90 and analyzed their effects on transcription and integrity of the TFIID and SAGA complexes. We found that the mutants displayed a variety of allele-specific defects in their ability to support transcription and maintain the structure of the TFIID and SAGA complexes. Sequencing of the alleles revealed that all have mutations corresponding to the C terminus of the protein, with most clustering within the conserved WD40 repeats; thus, the C terminus of TAF90p is required for its incorporation into TFIID and function in SAGA. Significantly, inactivation of one allele, taf90-20, caused the dramatic reduction in the levels of total mRNA and most specific transcripts analyzed. Analysis of the structure and/or activity of both TAF90p-containing complexes revealed that this allele is the most disruptive of all. Our analysis defines the requirement for the WD40 repeats in preserving TFIID and SAGA function, demonstrates that the defects associated with distinct mutations in TAF90 vary considerably, and indicates that TAF90 can be classified as a gene required for the transcription of a large number of genes.

Identification of charged residues in an N-terminal portion of the hormone-binding domain of the human estrogen receptor important in transcriptional activity of the receptor.

We have shown that charged amino acids near C530 of the human estrogen receptor (ER) are involved in receptor discrimination between estrogen and antiestrogen. We now examine the role of charged residues, adjacent to the three other cysteines (381, 417, and 447) in the hormone-binding domain of the human ER, in the hormone-binding, DNA-binding, and transcription activation abilities of the receptor. Mutation of the one charged amino acid nearest to C381 gave a mutant receptor (E380Q) requiring two to three times less estradiol (E2) than wild type (WT) ER to achieve maximal activity and having activity in the absence of added estrogen that was 6-fold higher than that of WT receptor. The enhanced ability of this mutant to bind to estrogen response element DNA in the absence and presence of estrogen may, at least in part, explain its elevated, seemingly constitutive trans-activation activity and its increased sensitivity to estrogen. While more sensitive to E2, this E380Q mutant was less sensitive than WT ER to antiestrogen for suppression of transcriptional activity. Mutation of all three charged residues nearest to C381 (the triple mutant D374N, E380Q, and E385Q) resulted in a greatly reduced potency of the receptor in trans-activation with no change in estrogen-binding affinity. When K449 (near C447), highly conserved among steroid receptors, was mutated to Q, 400-fold more E2 was required for maximal reporter gene trans-activation due to an unstable, temperature-sensitive hormone-receptor complex. In contrast, the mutant K416Q (near C417) was unaltered in E2-binding or receptor transcriptional activity. These studies reveal a region in the N-terminal portion of the hormone-binding domain (ca. amino acids 374-385) where alterations in charged residues result in either increases or decreases in receptor transcriptional activity with no change in receptor affinity for hormone. Our findings suggest that this region may be important in DNA binding and protein-protein interactions that modulate transcriptional activity of the ER. In addition, the region near C447, which is well conserved among steroid receptors, appears to be important in maintaining the receptor in a conformation that is stable at physiological (37 C) temperatures. To our knowledge, this is the first report of an ER (E380Q) with a sensitivity to E2 for trans-activation greater than that of WT receptor and having high trans-activation activity in the absence of added hormone.

Identification of two cysteines closely positioned in the ligand-binding pocket of the human estrogen receptor: roles in ligand binding and transcriptional activation.

We have previously identified cysteine 530 in the human estrogen receptor (ER) as the major site of attachment for covalently binding affinity ligands and have shown that when this cysteine is mutated to alanine (C530A mutant), the affinity ligand [tamoxifen aziridine (TAZ)] can still bind covalently to the ER, presumably by interaction with a different cysteine(s) in the hormone-binding domain (HBD). Using site-directed mutagenesis, we have determined the alternative ligand attachment site and the functional importance of the cysteines (residues 381, 417, 447, and 530) in the HBD of the ER to the hormone-binding and transcriptional responses to estrogens and antiestrogens. Cysteine 530 plus one or more of these other cysteines were mutated to alanines. Analysis of these mutant ERs expressed in Chinese hamster ovary cells provides strong evidence that cysteine 381 is the residue that is preferentially covalently labeled by TAZ in the C530A mutant. Hence, portions of the HBD that are far apart in the linear receptor sequence, namely regions near C381 and C530, are probably closely positioned in the ligand-binding pocket, with the cysteine thiols being 1.1 nm or less apart. The affinity of estradiol binding to receptors was reduced only 2- and 5-fold, respectively, in the double and quadruple Cys to Ala mutants, and estradiol was an effective stimulator of transcription from an estrogen-responsive reporter gene [(ERE)2-TATA-CAT].(ABSTRACT TRUNCATED AT 250 WORDS)

Categories of resistance to greenbug (Homoptera: Aphididae) biotype K in wheat lines containing Aegilops tauschii genes.

The wheat lines (cultivars) 'Largo', 'TAM110', 'KS89WGRC4', and 'KSU97-85-3' conferring resistance to greenbug, Schizaphis graminum (Rondani), biotypes E, I, and K were evaluated to determine the categories of resistance in each line to greenbug biotype K. Our results indicated that Largo, TAM110, KS89WGRC4, and KSU97-85-3 expressed both antibiosis and tolerance to biotype K. Largo, KS89WGRC4, and KSU97-85-3, which express antixenosis to biotype I, did not demonstrate antixenosis to biotype K. The results indicate that the same wheat lines may possess different categories of resistance to different greenbug biotypes. A new cage procedure for measuring greenbug intrinsic rate of increase (r(m)) was developed, by using both drinking straw and petri dish cages, to improve the efficiency and accuracy of r(m)-based antibiosis measurements.

Hepatic progesterone receptors: characterization in the turtle Chrysemys picta.

High and low affinity progesterone (P)-binding proteins similar to those described in the chick oviduct were characterized for the first time in liver cytosol in an oviparous turtle, Chrysemys picta. Two forms of high affinity P receptor were separated by diethyl aminoethyl (DEAE)-Sepharose anion exchange chromatography; the A form eluted in low salt buffer, and the B form at a KCl concentration of 0.20 M in a linear gradient. After photoaffinity labeling of crude cytosol with [3H] R5020, two overlapping peaks of radioactivity were detected with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After separation with DEAE-Sepharose and photoaffinity labeling, the A form migrated as a single peak with a mol wt of 90,000, and the B form as a single peak with a mol wt of 123,000. The affinity (Kd, 2.9 X 10(-9) M), capacity (1 pmol/mg protein), and binding specificity were characteristic of a P receptor. P competed most effectively for [3H]P binding. R5020 and deoxycorticosterone were good competitors; 5 alpha-pregnenolone, 17 alpha-hydroxyprogesterone, and testosterone were quite effective; cortisol, corticosterone, aldosterone, and 5 alpha-dihydrotestosterone were poor competitors. A lower affinity P-binding component (Kd, 1 X 10(-7) M; maximum binding, 15 pmol/mg protein) co-eluted from DEAE-Sepharose columns with the A and B receptor forms. This is the first description of a hepatic P receptor in any vertebrate.

Promoter targeting of chromatin-modifying complexes.

The action of multi-subunit complexes that are able to overcome the repressive effects of chromatin is an important step in the regulation of eukaryotic gene expression. Identification of complexes that modify the structure of chromatin to help factors access the underlying DNA has enhanced our understanding of how some genes are controlled. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) represent one group of complexes that regulate the level of acetylation on the N-terminal tails of core histone proteins. The SWI/SNF complex is the prototype of a second group of complexes, which use the energy of ATP-hydrolysis to alter histone-DNA contacts, leading to changes in chromatin conformation. Genetic studies in yeast have revealed that some of these multi-subunit complexes interact in vivo to control transcription of a subset of genes. It has become apparent that some gene promoters require modifications by both types of complexes. An important question regarding these two types of complexes is how they are recruited to the promoters of genes that are dependent on their activity for their expression. This review will tie together many studies on promoter recruitment of both HATs and SWI/SNF. Emphasis will be placed on recent data that demonstrates functional interplay between these two types of chromatin-remodeling activities. In addition, this review summarizes recent data demonstrating the ability of repressors and corepressors to recruit histone deacetylase complexes. Interestingly, many subunits of chromatin-modifying complexes in humans have been implicated in the development of cancer. Thus, studying how these complexes work can help us better understand human diseases.

The effect of FK506 on canine bile flow.

While the primary immunosuppressive agents utilized in solid-organ transplantation are steroids and cyclosporine, the more recently introduced agent FK506 is assuming a progressively more important role as an immunosuppressant, particularly in liver transplantation. While the effect of cyclosporine on rat and canine bile flow has been well evaluated, no information is available regarding the effect of FK506 on bile flow. Dogs with chronic biliary fistulas were utilized, enabling unanesthetized animals to be studied. FK506 was administered intravenously in varying doses, and bile volume, bile salts, and bile electrolytes were measured. FK506 produced dose-related increases in bile volume and bile chloride concentration and output, with 8 micrograms/kg-1hr-1 being the maximal dose. To ascertain that the response was not osmotic in association with FK506 secretion in bile, 500 micrograms/kg-1hr-1 FK506 was administered, which did not produce a choleresis significantly greater than 8 micrograms/kg-1hr-1. FK506 was subsequently administered orally in daily dose of 0.15 mg/kg-1 for two weeks. Oral FK506 did not consistently increase bile flow, as evaluated by a bile salt dose-response curve (9, 18, 36 mumol/min sodium taurocholate) but did significantly increase bile chloride secretion. Two weeks of oral administration of FK506 in therapeutic doses did not significantly alter serum bile salt concentrations. The results of this study indicate that intravenous FK506 produces a chloride-rich choleresis in dogs.

The effect of octreotide acetate on meal-stimulated exocrine secretion in canine pancreatic autografts.

Octreotide acetate (Sandostatin), a long-acting somatostatin analogue, has been demonstrated to have an inhibitory effect on exocrine secretion in the neurally intact pancreas. This study was designed to evaluate the effect of this agent on exocrine secretion in the denervated canine pancreas, utilizing animals with pancreatic autografts and functioning pancreaticocystostomies. The rates of secretion of urinary (autograft) amylase (units/min) and bicarbonate (mM/min), over a five-hr interval, were determined in the basal state (group A, n = 10), after a bolus injection of 400 micrograms of Sandostatin (group B, n = 5), after a standard meal (group C, n = 5), or a meal preceded by 400 micrograms of Sandostatin (group D, n = 5). Basal secretion of amylase was decreased for 4 hr following Sandostatin, although this decrease was not significant. Conversely, basal bicarbonate secretion was not inhibited by Sandostatin. When compared with group C (22.4 +/- 3.2), a significant inhibition of meal-stimulated amylase release was demonstrated in group D (5.4 +/- 0.21, P = 0.0006) during the first hour after Sandostatin was given. This inhibition remained significant at 2 hr (group C = 38.5 +/- 5.2 versus group D = 9.4 +/- 0.8; P = 0.0006) and 3 hr (group C = 38.6 +/- 6.3 versus group D = 17.5 +/- 0.9; P = 0.0108) after Sandostatin was given. In addition, meal-stimulated bicarbonate secretion was significantly inhibited for 2 hr following Sandostatin (group C = 0.19 +/- 0.03 versus group D = 0.07 +/- 0.02, P = 0.0096; and group C = 0.23 +/- 0.03 versus group D = 0.10 +/- 0.01, P = 0.0018, respectively). These studies demonstrate that Sandostatin has a profound inhibitory effect on meal-stimulated enzyme and bicarbonate release in a denervated canine autograft model. Although the site of action of this agent remains to be defined, Sandostatin may have therapeutic potential in clinical pancreas transplantation.

Hormone binding and transcription activation by estrogen receptors: analyses using mammalian and yeast systems.

We have used affinity labeling, site-directed mutagenesis and regional chemical mutagenesis in order to determine regions of the human estrogen receptor (ER) important in hormone binding, ligand discrimination between estrogens and antiestrogens, and transcriptional activation. Affinity labeling studies with the antiestrogen, tamoxifen aziridine and the estrogen, ketononestrol aziridine have identified cysteine 530 in the ER hormone binding domain as the primary site of labeling. In the absence of a cysteine at 530 (i.e. C530 mutant), C381 becomes the site of estrogen-compatible tamoxifen aziridine labeling. Hence these two residues, although far apart in the primary linear sequence of the ER protein, must be close in the three-dimensional structure of the protein, in the ER ligand binding pocket, so that the ligand can reach either site. Site-directed mutagenesis of selected residues in the ER and region-specific chemical mutagenesis of the ER hormone binding domain with initial phenotypic screening in yeast have enabled the identification of a region near C530 important in discrimination between estrogens and antiestrogens and of other residues important in hormone-dependent transcriptional activation. Some ER mutants with alterations in the carboxy-terminal portion of the hormone binding domain are transcriptionally inactive yet bind hormone and also function as potent dominant negative ERs, suppressing the activity of wild-type ER at low concentrations. These studies reveal a separation of the hormone binding and transcription activation functions of the ER. They are also beginning to provide a more detailed picture of the ER hormone binding domain and amino acids important in ligand binding and discrimination between different categories of agonist and antagonist ligands. Such information will be important in the design of maximally effective antiestrogens. In addition, since there is now substantial evidence for a mixture of wild-type and variant ERs in breast cancers, our studies should provide insight about the bioactivities of these variant receptors and their roles in modulating the activity of wild type ER, and should lead to a better understanding of the possible role of variant receptors in altered response or resistance to antiestrogen and endocrine therapy in breast cancer. In addition, some dominant negative receptors may prove useful in examining ER mechanisms of action and in suppressing the estrogen-dependent growth of breast cancer cells.

Splenopneumopexy to treat portal hypertension produced by venous occlusive disease.

Splenopneumopexy is intended to induce collateral circulation between the portal system and the pulmonary veins. It involves performing a parenchymatous anastomosis between the amputated superior pole of the spleen and the exposed pulmonary venous structures in the left lower lobe. This operative procedure was used to treat four patients with extended portal-splenic-mesenteric venous occlusion who did not respond satisfactorily to sclerotherapy. The patients underwent transfemoral embolization of their splenic arteries before splenopneumopexy. Following the operations, all patients have remained well, experiencing cessation of gastrointestinal bleeding for up to 48 months. Splenopneumopexy may be a therapeutic alternative in selected patients with portal hypertension, including those patients with widespread occlusion of the portal vein and its radicles.

Effect of somatostatin and octreotide acetate on OP-CCK-stimulated exocrine secretion in the denervated canine pancreas.

Somatostatin and its analogue, octreotide acetate (Sandostatin), have been demonstrated to suppress exocrine secretion in a denervated canine pancreatic autograft model. To help define this inhibitory mechanism, the effect of these agents on cholecystokinin (CCK)-stimulated acinar cell secretion was evaluated. In vitro assessment evaluated the effect of somatostatin on octapeptide (OP)-CCK-stimulated amylase release of pancreatic tissue slices. In vivo assessment employed animals with pancreatic autografts and pancreaticocystostomies, evaluating the effect of a bolus intravenous injection of 100 micrograms of octreotide acetate on the basal and OP-CCK-stimulated (125 ng/kg/h) secretion of urinary (autograft) amylase and bicarbonate. Incubation of tissue slices with 0.16, 0.24, or 0.32 microgram/ml somatostatin had no significant effect on in vitro OP-CCK-simulated amylase release. Intravenous octreotide acetate resulted in a significant decrease in the basal rate of amylase secretion but had no significant effect on OP-CCK-stimulated autograft amylase or bicarbonate release. These studies demonstrate that octreotide acetate has an in vivo inhibitory effect on basal amylase release of pancreatic autografts but cannot counteract maximal stimulation with exogenous OP-CCK. Also, somatostatin does not inhibit OP-CCK-stimulated acinar cell secretion of pancreatic tissue slices. These results indicate that the exocrine inhibition produced by somatostatin analogues in the grafted pancreas occurs via an indirect mechanism.

Anaphylactoid reaction to muromonab-CD3 in a pediatric renal transplant recipient.

Muromonab-CD3 (OKT3), a murine IgG2a antibody directed against the T3 (CD3) complex on mature lymphocytes, triggers adverse immune reactions. Anaphylactic reactions have occurred in patients exposed to OKT3 and are mediated by anti-OKT3 IgE antibodies. The reactions are not antibody mediated and can occur within seconds of administration of a mast cell secretogogue. A renal transplant recipient became hypotensive and hypoxic immediately after receiving her first dose of OKT3 and required advanced life support. Serum antibody tests were negative for anti-OKT3 IgG, IgE, and antimouse protein antibodies. To our knowledge, this is the first published report of a patient with an anaphylactoid reaction to the initial infusion of OKT3.

Categories of resistance to greenbug (Homoptera: Aphididae) biotype I in Aegilops tauschii germplasm.

Categories of resistance to greenbug, Schizaphisgraminum (Rondani), biotype I, were determined in goatgrass, Aegilops tauschii (Coss.) Schmal., accession 1675 (resistant donor parent), 'Wichita' wheat, Triticum aestivum L., (susceptible parent), and an Ae. tauschii-derived resistant line, '97-85-3'. Antibiosis was assessed using the intrinsic rate of increase (rm) of greenbugs confined to each of the three genotypes. Neither parent nor the resistant progeny expressed antibiosis. Mean rm values for greenbug I on Wichita (0.0956), and Ae. tauschii (0.10543) were not significantly different. Mean rm values for Wichita and 97-85-3 were also not significantly different. Antixenosis was determined by allowing aphids a choice to feed on plants of each of the three genotypes. Ae. tauschii 1675 exhibited antixenosis, but this resistance was not inherited and expressed in '97-85-3'. In experiments comparing Wichita and Ae. tauschii 1675, greenbug I population distributions were not significantly different on Wichita at 24 h, but were shifted toward Wichita at 48 h. In the second antixenosis experiment, there were no significant differences in greenbug I population distributions on 97-85-3 or Wichita at 24 or 48 h. When all three lines were compared, there were no significant differences in greenbug biotype I populations at 24 or 48 h after infestation. Comparisons of proportional dry plant weight loss (DWT) and SPAD meter readings were used to determine tolerance to greenbug I feeding. Ae. tauschii 1675 and 97-85-3 were highly tolerant compared with Wichita. Infested and uninfested Ae. tauschii 1675 DWT was nonsignificant, and infested Wichita plants weighed significantly less than uninfested plants. When Wichita and 97-85-3 were contrasted, DWT of infested and uninfested Wichita plants were significantly different, but those of 97-85-3 were not. Mean percent leaf chlorophyll losses for the three genotypes, as measured by the SPAD chlorophyll meter, were as follows: Wichita = 65%; Ae. tauschii 1675 = 25%; and 97-85-3 = 39%. Percent leaf chlorophyll losses caused by greenbug feeding was significantly different in comparisons between Wichita and Ae. tauschii 1675, and comparisons between Wichita and 97-85-3, although feeding damage was not significantly different in comparisons between Ae. tauschii 1675 and 97-85-3. These data provided further evidence of the expression of tolerance to greenbug feeding in Ae. tauschii 1675 and 97-85-3.