Rhesus monkey micro mixed lymphocyte and mitogen reactivity: Optimal conditions and normal variability

Optimal conditions for the rhesus monkey micro mixed lymphocyte system with multiple automated harvesting of samples were evaluated. Parameters studied were cell concentration, length of culture period, methods of inactivation of cell populations, supplementation of media, type of culture plates, and changes in the reactivity of cells from individual animals over an extended time period. This work was supported in part by Portland Veterans Administration Hospital, Portland, Oregon, and the General Research Support Branch of the U.S. Public Health Service Grant RR00163, the Bureau of Medicine and Surgery, Navy Department, Work Unit No. M4318. 01.007ABG2. The opinions or assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the U.S. Navy Department or the Naval service at large. The animals used in this study were handled in accordance with the provisions of Public Law 89–54 as amended by Public Law 91–579, “Animal Welfare Act of 1970,” and the principles outlined in the “Guide for the Care of Laboratory Animals,” U.S. Department of Health, Education, and Welfare Publication No. (NIH) 73-23.     

Formation of ATP by the adenosine triphosphatase complex from spinach chloroplasts reconstituted together with bacteriorhodopsin

The energy-linked ATPase complex has been isolated from spinach chloroplasts. This protein complex contained all the subunits of the chloroplast coupling factor (CF₁) as well as several hydrophobic components. When the activated complex was reconstituted with added soybean phospholipids, it catalyzed the exchange of radioactive inorganic phosphate with ATP. Sonication of the complex into proteoliposomes together with bacteriorhodopsin yielded vesicles that catalyzed light-dependent ATP formation. Both the ³²P_i-ATP exchange reactions and ATP formation were sensitive to uncouplers such as 3-tert-butyl-5,2′-dichloro-4′-nitrosalicylanilide, bis-(hexafluoroacetonyl)acetone and carbonyl cyanide-p-trifluoromethoxyphenyl-hydrazone, that act to dissipate a proton gradient. The energy transfer inhibitors dicyclohexylcarbodiimide, triphenyltin chloride and 2-β-d-glucopyranosyl-4,6′-dihydroxydihydrochalcone were also effective inhibitors of both reactions.     

Longitudinal patterns of production, food consumption, and seston utilization by net-spinning caddisflies (Trichoptera) in a southern Appalachian stream (USA)

Larval production of ten species of Hydropsychidae and Philipotamidae was studied at six stations along 6.4 km of a southern Appalachian stream, encompassing stream orders 1–4 and a 600 m elevation change. Species-specific production estimates ranged from 23–983 mg AFDM m^?2 yr^?1 These low values are attributed to the paucity of nutrients in these undisturbed headwater streams which reduces detrital food quality, algal growth, and production of smaller invertebrates eaten by hydropsychids. Animal food supported the majority of hydropsychid production (72%); philopotamids relied primarily on fine detritus (80%) and diatoms (15%). The contribution of animal food to caddisfly production decreased downstream, while the relative importance of filamentous algae and diatoms increased. These changes reflect the downstream decline of more carnivorous species, as well as increased primary production which accompanies the shift in lotic community metabolism from heterotrophy towards autotrophy with increasing stream order. Net-spinning caddisflies had a minor impact on seston quantity, consuming only 0.0003%-0.005% of the total seston (including invertebrate drift) passing over a m² of substrate annually. In contrast, the percentage of invertebrate drift consumed was, on the average, > 400 × higher than total seston consumption. These insects influence seston quality rather than quantity. The percentage of total seston and animal drift consumed declined downstream, indicating that turnover lengths of these materials increase with stream order. Longer turnover lengths or “spirals” result from changes in the physical characteristics of the stream, i.e., increasing discharge and stream power and decreasing numbers of retention devices (i.e., organic debris dams), which increase the downstream transport velocity of seston. Higher transport velocities reduce the rates at which these filter feeding caddisflies can process the organic inputs to a given reach of stream. Small streams (orders 1–3) appear to be most responsible for efficient processing of a stream's energy inputs.     

Purification and properties of soybean nodule xanthine dehydrogenase

Xanthine dehydrogenase (EC 1.2.1.37), an essential enzyme for ureide metabolism was purified from the cytosol fraction of soybean nodules. The purified xanthine dehydrogenase was shown to be homogeneous by electrophoresis and a pI of 4.7 was determined by isoelectric focusing. The enzyme had a molecular weight of 285,000 and two subunits of molecular weight 141,000 each. The holoenzyme contained 1.7 (±0.7) mol Mo and 8.1 (±2.0) mol Fe/mol enzyme and the enzyme also contained FMN and is thus a molybdoironflavoprotein. Soybean xanthine dehydrogenase is the second enzyme in plants demonstrated to contain Mo and the first xanthine-oxidizing enzyme reported to contain FMN, rather than FAD as the flavin cofactor.     

Plasmid Involvement in Parathion Hydrolysis by Pseudomonas diminuta

An organism identified as Pseudomonas diminuta was found to hydrolyze parathion. Cells grown for 48 h contained 3,400 U of parathion hydrolase activity per liter of broth. Expression of enzymatic activity was lost at a high frequency (9 to 12%) after treatment with mitomycin C. Hydrolase-negative derivatives were missing a plasmid present in the wild-type organism. The molecular mass of this plasmid (pCS1), as determined by electron microscopy, was about 44 × 10⁶ daltons.     

Early events in calcium-induced liposome fusion

Calcium-induced interaction of liposomes composed of pure phosphatidylserine (PS) has been studied using a rapid-mixing, rapid-freeze device. Freeze-fracture electron microscopy of this material revealed that liposomes react very rapidly after addition of calcium ions. After only 10 ms (the resolution of the technique) vesicle fusion was apparent. At the same time, however, vesicles also collapsed, and appeared as aggregates of flattened membranes. This may explain controversies which have arisen over vesicle fusion studied with more indirect methods.     

A cytological basis for the separation of Geropogon from Tragopogon (Compositae: Lactuceae)

The single species ofGeropogon, G. hybridus (L.) Schultz-Bip., is sometimes submerged in the morphologically similarTragopogon. A comparative study of the karyotypes ofGeropogon andTragopogon sensu stricto clarified their relationship. The karyotype ofG. hydridus (2n=14) was compared with that ofT. dubius Scop. (2n=12). The karyotypes are distinct, offering a compelling reason to recognizeGeropogon as a separate genus.     

Effect of food-deprivation on rat pancreatic islet cells

Somatostatin, insulin and glucagon concentrations in rat pancreas were measured following various intervals of food-deprivation. Tissue concentrations, as measured by radioimmunoassay, were correlated with A-, B-, and D-cell number and size using a scanning integrating image analyzer (Quantimet 720). Alterations in total islet hormone content were not correlated to changes in size or distribution of cells. This implies that changes in tissue content reflect changes in turnover of peptides rather than changes in cell size or number.