Platte Valley Yellows,a chlorotic condition of soybeans: Symptoms and preliminary chemical analyses

Summary A chlorotic condition of soybeans in the river valleys of eastern Nebraska was studied. The pattern of chlorosis, presence of petiolar necrosis, reductions in rhizobial and mycorrhizal infection, and plant and soil chemical analyses suggest that the condition is more complex than normal iron deficiency chlorosis or other mineral deficiencies or toxicities. Nodulation is almost totally inhibited in affected plants. Mycorrhizal fungi, while able to infect early in the season, do not persist on affected plants. Breakdown of endodermis was observed in the roots of affected plants. The factors contributing to this condition have not yet been determined. Published with the approval of the director as Paper No. 5672, Journal Series, Nebraska Agricultural Experiment Station.     

α-d-allopyranosyl β-d-fructofuranoside (allo-sucrose) and its derivatives

Reduction of 3-ketosucrose (1) with sodium borohydride gave mainly α-d-allopyranosyl β-d-fructofuranoside (2) characterized as its octabenzoate. Using sodium borodeuteride, [3-²H]allo-sucrose (5) and [3-²H]sucrose (6) were obtained in the ratio 12:1. The mixture was fractionated on Dowex-50 X8 resin (Ca²⁺ form), and the [3-²H] derivatives were isolated as their octa-acetates. Inspection of the ¹³C-n.m.r. spectra of 5 and 6 enabled the C-3 signals to be assigned. allo-Sucrose (2) was more readily obtained by oxidation of sucrose with dimethyl sulphoxide-acetic anhydride followed by reduction with sodium borohydride and fractionation on Dowex-50 X8 (Ca²⁺) resin. Tritylation of 2 followed by acetylation gave, after chromatography, the 6,1′,6′-tritrityl ether (9, 10%), the 6,6′-ditrityl ether (10, 26%), and a mixture of monotrityl ethers (20%). Hydrogenolysis of 9 and 10 gave the penta-acetate and hexa-acetate, respectively, with no detectable migration of AcO-4. Treatment of 2 with sulphuryl chloride at -50° gave the 6,6′-dichloride.     

Fibronectin–collagen binding and requirement during cellular adhesion

Fibronectin isolated from human plasma and from the extracellular matrices of cell monolayers mediates the attachment in vitro and spreading of trypsin-treated cells on a collagen substratum. Fibronectin-dependent kinetics of cellular attachment to collagen were studied for several adherent cell types. It was shown that trypsin-treated human umbilical-cord cells, mouse sarcoma CMT81 cells, endothelial cells, and human fibroblasts from a patient with Glanzmann's disease were completely dependent on fibronectin for their attachment to collagen, whereas guinea-pig and monkey smooth-muscle cells and chick-embryo secondary fibroblasts displayed varying degrees of dependence on fibronectin for their attachment. Radiolabelled human plasma fibronectin possessed similar affinity for collagen types I, II and III from a variety of sources. The fibronectin bound equally well to the collagens with or without prior urea treatment. However, in the fibronectin-mediated adhesion assay using PyBHK fibroblasts, a greater number of cells adhered and more spreading was observed on urea-treated collagen. Fibronectin extracted from the extracellular matrix of chick-embryo fibroblasts and that purified from human plasma demonstrated very similar kinetics of complexing to collagencoated tissue-culture dishes. Fibronectin from both sources bound to collagen in the presence of 0.05–4.0m-NaCl and over the pH range 2.6–10.6. The binding was inhibited when fibronectin was incubated with 40–80% ethylene glycol, the ionic detergents sodium dodecyl sulphate and deoxycholate, and the non-ionic detergents Nonidet P-40, Tween 80 and Triton X-100, all at a concentration of 0.1%. From these results we proposed that fibronectin–collagen complexing is mainly attributable to hydrophobic interactions.     

The formation of dipeptides from amino acids and the 2′(3′)-glycyl ester of an adenylate

Summary The yields of dipeptide obtained from the reaction of 0.2M 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) and 0.2M amino acid at pH 8.2 ranged from 0.1% to 35.5% for a group of 15 amino acids. The yields of glyser (35.3%), gly-cys (11.8%) and gly-thr (5.4%) were considerably greater than dipeptide yields obtained from any of the other 12 amino acids ( 1.7%). Aminolysis of 0.05M 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) by 0.4M serine ethyl ester yielded 53% glycylserine diketopiperazine, via N-(glycyl)-serine ethyl ester as a transient intermediate. The prebiotic significance of these reactions is discussed.Abbreviations MepA adenosine-5-(O-methylphosphate) - MepA-gly 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) - DKP diketopiperazine - serOEt serine ethyl ester - gly-serOEt N-(glycyl)-serine ethyl ester - Boc-gly N-tertbutyloxycarbonylglycine - cyclo-(gly-ser-) glycylserine diketo-piperazine - O-gly-ser O-glycylserine - O-(gly)-gly-ser O-(glycyl)-glycylserine - gly-ser N-glycylserine     

Circular dichroism of human urinary Tamm-Horsfall glycoprotein

Summary Human urinary Tamm-Horsfall glycoprotein, which contains 28% carbohydrate, has a monomeric molecular weight of about 80,000 but is isolated from urine in the form of intertwining helical suprastructures with molecular weights greater than 10⁷. The native glycoprotein was dissociated and denatured with 6 M guanidinium chloride and was subsequently renatured by dialysis against a Tris-HCl buffer. Using sedimentation equilibrium, the renatured glycoprotein was characterized by a of 256,800 and a of 356,000. The ratio,M _z/M _w, of 1.39 indicates some polydispersity with regard to molecular size. There was no evidence of helical suprastructures in the renatured glycoprotein as judged by electron microscopy. Ca²⁺ concentrations of up to 50 mM failed to precipitate the renatured glycoprotein; in contrast, the native glycoprotein is precipitated by Ca²⁺ concentrations between 5–10 mM. The circular dichroic spectrum of renatured Tamm-Horsfall glycoprotein was obtained, resolved, and tentative band assignments made. The spectrum, which is quite similar to that of native Tamm-Horsfall glycoprotein, exhibited negative extrema at 269 nm (due in large part to disulfides and tyrosines) and at 215 nm (due to protein-structure and the N-acetylated hexosamines). The-helical content of the glycoprotein was estimated to be no more than 10% and the amount of-structure to be about 33%; these values were not affected by the presence of Ca²⁺ (1 mM). A glycopeptide fraction (ca. 90% carbohydrate), prepared by extensive pronase digestion of the reduced, S-carboxymethylated glycoprotein, exhibited an ellipticity extremum at 212 nm of +4,750 deg · cm²/dmole, referred to the concentration of (N-acetylated) hexosamines and neuraminic acid.Research Career Development Awardee (AM-00055).     

Augmented therapeutic results elicited by splenectomy in combined modality treatment of spontaneous murine breast cancer

Summary Because it had been reported that splenectomy produces a tumor-inhibitory effect in several transplantable tumor systems when the surgery is performed before tumor challenge, we attempted to examine this putative immunological manipulation in a therapeutic situation.A spontaneous, autochthonous, murine breast tumor system was utilized in the present studies, and treatment was initiated in animals bearing large tumors (averaging 0.5 g). To amplify any immunological benefit ensuing from splenectomy, the tumor burden in the host was reduced by ancillary treatment with enucleative tumor surgery or with enucleative tumor surgery plus cytoreductive combination chemotherapy.Splenectomy performed in conjunction with enucleative tumor surgery was associated with an increment of cure in each of four separate experiments in comparison to treatment with enucleative tumor surgery alone. In four of five experiments utilizing different combinations or schedules of chemotherapeutic agents following enucleative tumor surgery, the addition of splenectomy resulted in a decrease in the rate of tumor recurrence as well as an increment in the cure rate. In the fifth experiment, splenectomy resulted in a decrease in the rate of tumor recurrence, but did not effect the ultimate cure rate.Although the nature of the immunological changes resulting from splenectomy are incompletely defined at present, these results provide encouragement in the search for immunological treatments for solid tumors.This work was supported in part by Contract No. N01-CM-73703 and Grant IR01CA-14768-01A1, both from the National Cancer Institute, National Institutes of Health, Department of Health, Education and Welfare, USA, and in part by a grant from the Chemotherapy Foundation of New York, Inc.     

A METHOD OF CORONARY RETROPERFUSION FOR THE TREATMENT OF ACUTE MYOCARDIAL ISCHEMIA

A method of retrograde perfusion of the myocardium has been developed in dogs. It consists of a double lumen balloon-tipped catheter inserted transvenously into the coronary sinus, with one lumen connected to a roller pump, the other to a helium counterpulsing pump. Oxygenated heparinized blood is obtained from the femoral artery and pumped continuously into the coronary sinus at a pressure of 50-75 mm Hg. The balloon is inflated during diastole, sealing the coronary sinus and promoting retrograde flow, and is deflated during systole, allowing blood drainage into the right atrium and preventing venous congestion. Thirteen anesthetized open-chest dogs were subjected to 15 minutes of proximal LAD artery occlusion and 30 minutes of diastolic coronary sinus perfusion (DCSP). The area of ischemia was mapped by means of platinum electrodes capable of simultaneously measuring myocardial tissue oxygen tension M(p)O(2)) and electrograms. Reduction of M(p)O(2) with simultaneous elevation of the ST segment on the corresponding electrogram was considered an indication of ischemia. Diastolic coronary sinus perfusion improved myocardial oxygen tension in the ischemic myocardium, reduced ST segment elevation, and tended to restore arterial blood pressure. Histologically, there was no intramyocardial hemorrhage.     

Single and multiple turnover reactions in the ubiquinone-cytochrome b-c2 oxidoreductase of Rhodopseudomonas sphaeroides. The physical chemistry of the major electron donor to cytochrome c2, and its coupled reactions

We have examined the thermodynamic properties of the physiological electron donor to ferricytochrome c₂ in chromatophores from the photosynthetic bacterium Rhodopseudomonas sphaeroides. This donor (Z), which is capable of reducing the ferri-cytochrome with a halftime of 1–2 ms under optimal conditions, has an oxidation-reduction midpoint potential of close to 150 mV at pH 7.0, and apparently requires two electrons and two protons for its equilibrium reduction.

The state of reduction of Z, which may be a quinone · protein complex near the inner (cytochrome c₂) side of the membrane, appears to govern the rate at which the cyclic photosynthetic electron transport system can operate. If Z is oxidized prior to the flash-oxidation of cytochrome c₂, the re-reduction of the cytochrome takes hundreds of milliseconds and no third phase of the carotenoid bandshift occurs. In contrast if Z is reduced before flash activation, the cytochrome is rereduced within milliseconds and the third phase of the carotenoid bandshift occurs. The prior reduction of Z also has a dramatic effect on the uncoupler sensitivity of the rate of electron flow; if it is oxidized prior to activation, uncoupler can stimulate the cytochrome re-reduction after several turnovers by less than tenfold, but if it is reduced prior to activation, the stimulation after several turnovers can be as dramatic as a thousandfold. The results suggest that Z plays a central role in controlling electron and proton movements in the ubiquinone cytochrome b-c₂ oxido-reductase.