KCl loss and cell shrinkage in the ehrlich ascites tumor cell induced by hypotonic media, 2-deoxyglucose and propranolol

Ehrlich ascites tumor cells lose KCl and shrink after swelling in hypotonic media and in response to the addition of 2-deoxyglucose, propranolol, or the Ca²⁺ ionophore, A23187, plus Ca²⁺ in isotonic media. All of these treatments activate cell shrinkage via a pathway with the following characteristics: (1) the KCl loss responsible for cell shrinkage does not alter the membrane potential; (2) NO₃^? does not substitute for Cl^?; (3) the net KCl movements are not inhibited by quinine or DIDS; and (4) early in this study furosemide was effective in inhibiting cell shrinkage but this sensitivity was subsequently lost. This evidence suggests that the KCl loss in these cells occurs via a cotransport mechanism. In addition, hypotonic media and the other agents used here stimulate a Cl^? -Cl^? exchange, a net loss of K⁺ and a net gain of Na⁺ which are not responsible for cell shrinkage. The Ehrlich cell also appears to have a Ca²⁺-activated, quinine-sensitive K⁺ conductive pathway but this pathway is not part of the mechanism by which these cells regulate their volume following swelling or shrink in isotonic media in response to 2-deoxyglucose or propranolol. Shrinkage by the loss of K⁺ through the Ca²⁺ stimulated pathway appears to be limited by Cl^? conductive movements; for when NO₃^?, an anion demonstrated here to have a higher conductive movement than Cl^?, is substituted for Cl^?, the cells will shrink when the Ca²⁺-stimulated K⁺ pathway is activated.     

The role of the pituitary-adrenal axis in the hyperthermia induced by acute peripheral or central (preoptic anterior hypothalamus) administration of morphine to unrestrained rats

The role of the pituitary-adrenal axis in the mediation of morphine-induced hyperthermia of conscious, unrestrained rats was investigated. Rectal (TR) and tail (Tt) temperatures and oxygen uptake rates (VO2) were measured following peripheral or central injection of morphine sulphate (MS) in groups of Sprague-Dawley rats before and after adrenalectomy (adx), hypophysectomy (hyp), or pituitary suppression (via dexamethasone treatment). The hyperthermic TR responses of groups given MS either subcutaneously (5 or 15 mg/kg) or directly into the preoptic anterior hypothalamus (POAH, 1 or 10 micrograms/microL) before adx were not different upon retesting with the same dose of MS 2 weeks later following adx. The hyperthermia with MS was not caused by vasoconstriction or by increases in basal metabolic rate, for Tt rose after the opiate injections whereas oxygen uptake rates (VO2) were reduced. Unexpectedly, the TR following POAH injections of sterile saline (SS) or deionized water after adx increased from those seen before adx. Adx groups supplemented with dexamethasone phosphate (either chronically with 20 micrograms/kg daily for 2 weeks post-adx before retesting with MS or acutely with 250 micrograms/kg 2 h before retesting) showed a hyperthermia to MS (5 mg/kg sc or 1.0 microgram/microL POAH) similar to that seen before adx. However, dexamethasone phosphate (250 micrograms/kg) supplementation to adx rats, that received POAH injections of SS, did reduce the rise in TR. Hyp rats given MS (5 mg/kg, sc) also evoked hyperthermic responses similar to those of non-hyp control groups. The results clearly show that the acute hyperthermia of unrestrained rats induced by either peripheral or central injections of morphine is not caused by activation of the pituitary-adrenal axis.     

Visualization by Immune Electron Microscopy of a 27-nm Particle Associated with Acute Infectious Nonbacterial Gastroenteritis

A 27-nm particle was observed by immune electron microscopy in an infectious stool filtrate derived from an outbreak in Norwalk, Ohio, of acute infectious nonbacterial gastroenteritis. Both experimentally and naturally infected individuals developed serological evidence of infection; this along with other evidence suggested that the particle was the etiological agent of Norwalk gastroenteritis.     

Cell division in the olfactory epithelium of the lamprey,Lampetra fluviatilis

Summary The turnover of cells within the olfactory epithelium of the lamprey Lampetra fluviatilis was investigated using tritiated thymidine followed by autoradiography. It was found that cell division occurred in three distinct regions of the olfactory lamellae. Two of these regions — a distal lamellar region and a proximal lamellar region occurred outside the sensory area proper, but appeared to contribute cells to the sensory area as well as giving rise to secretory or ciliated cells outside the sensory area. A third region of division occurrred at the base of the sensory area. Division of specialised basal or blastema cells in this region gives rise to cells that are confined to the sensory region of the lamellae. These findings are discussed in the light of previous studies on cell replacement within the olfactory epithelium.     

Suppression of food intake and body weight gain by naloxone in rats

The effect of acute and chronic administration of naloxone on food acquisition and weight gain in rats was studied in 3 experiments. One injection of a sparingly-soluble salt of naloxone in slow-release vehicle markedly lowered mean food intake over that of control rats injected with the vehicle only. Mean body weight of the naloxone-injected rats was significantly lower than that of the control group for one week.Repeated evening injections (2000 h) of naloxone hydrochloride in saline tended to reduce the night-time feeding below control levels throughout the 10-day period of naloxone administration. Food intake was significantly lower in the 4- and 8-h periods after the first injection of naloxone than that on the preceding saline control night. The initial decreases were offset by increased day-time feeding so that total daily food intake was not significantly altered over the 10 days. When saline was substituted for naloxone, food intake increased.Rats given naloxone following 24 h of fasting consumed significantly less food and gained less weight during 4 h of access to food compared to those receiving saline. After a 48-h fast naloxone-treated rats also gained significantly less body weight than those given saline, but the reduction in food intake was not statistically significant. These results suggest the possibility that endorphins may have a modulating effect on feeding activity.     

Actin filaments elongate from their membrane-associated ends

In limulus sperm an actin filament bundle 55 mum in length extends from the acrosomal vacuole membrane through a canal in the nucleus and then coils in a regular fashion around the base of the nucleus. The bundle expands systematically from 15 filaments near the acrosomal vacuole to 85 filaments at the basal end. Thin sections of sperm fixed during stages in spermatid maturation reveal that the filament bundle begins to assemble on dense material attached to the acrosomal vacuole membrane. In micrographs fo these early stages in maturation, short bundles are seen extending posteriorly from the dense material. The significance is that these short, developing bundles have about 85 filaments, suggesting that the 85-filament end of the bundle is assembled first. By using filament bundles isolated and incubated in vitro with G actin from muscle, we can determine the end “preferred” for addition of actin monomers during polymerization. The end that would be associated with the acrosomal vacuole membrane, a membrane destined to be continuous with the plasma membrane, is preferred about 10 times over the other, thicker end. Decoration of the newly polymerized portions of the filament bundle with subfragment 1 of myosin reveals that the arrowheads point away from the acrosomal vacuole membrane, as is true of other actin filament bundles attached to membranes. From these observations we conclude that the bundle is nucleated from the dense material associated with the acrosomal vacuole and that monomers are added to the membrane-associated end. As monomers are added at the dense material, the thick first-made end of the filament bundle is pushed down through the nucleus where, upon reaching the base of the nucleus, it coils up. Tapering is brought about by the capping of the peripheral filaments in the bundle.     

Closed-loop control of fed-batch cultures of recombinant Escherichia coli using on-line HPLC

This article describes a fully automated system for the on-line monitoring and closed-loop control of a fed-batch fermentation of recombinant Escherichia coli, and presents two case studies of its used in limiting production of unwanted byproducts such as acetic in fed-batch fermentations. The system had two components. The first components, on-line monitoring, comprised an aseptic sampling device, a microcentrifuge, and HPLC System. These instruments removed a Sample from a fermentor, spun it at high speed to separate solid and liquid components, and then automatically injected the supernatant onto an HPLC column for analysis. The second component consisted of control algorithms programmed using the LabView visual programming environment in a control computer that was linked via a remote components were linked so that results from the on-line HPLC were captured and used by the control algorithm was designed to demonstrate coarse feedback control to confirm the operability of the controller. The second case study showed how the system could be used in a more sophisticated feedings strategy providing fine control and limiting acetate concentration to a low level throughout the fermentation. (c) 1994 John Wiley & Sons, Inc.     

The notal organ of the scorpionfly (Panorpa vulgaris): an adaptation to coerce mating duration

The notal organ in P. vulgaris (Insecta: Mecoptera) is a clamplikestructure on the dorsum of the middle of the male’s abdomenthat holds one of the female’s forewings throughout mating.Males often provide salivary masses as food to their mates duringmating, and a male’s ability to produce saliva dependsupon whether he has fed adequately before mating. To test thehypothesis that the evolutionary function of the notal organis to coerce longer matings than are in the interests of females,the notal organ and the amount of food males and females receivebefore mating were experimentally manipulated. In support ofthe hypothesis, copulation was reduced when the notal organwas made inoperative under the following four conditions: (1)female fed and male starved before mating, no saliva duringmating; (2) same as in condition 1 but with one salivary massduring mating; (3) both sexes starved before mating, no salivarymass during mating; and (4) both sexes fed before mating. However,when females were starved before mating and males were fed,resulting in multiple masses being provided during mating, thenotal organ had no effect on length of copulation, and thusthere was no sexual conflict over mating duration. In addition,the larger the female relative to her mate, the briefer thecopulation when the notal organ was operative, which suggeststhat a physical struggle between the male and female may occurduring sexual conflict about mating duration. This is one ofonly a few studies that provide evidence of adaptation to thedomain of sexual coercion. [Behav Ecol 1991;2:156–164]     

IL-4 increases human endothelial cell adhesiveness for T cells but not for neutrophils

The adhesion of leukocytes to vascular endothelium is the first step in their passage from the blood into inflammatory tissues. By modulating endothelial cell (EC) adhesiveness for leukocytes, cytokines may regulate leukocyte accumulation and hence the nature and progression of inflammatory responses. We have found that the T cell cytokine IL-4 increases the adhesion of T cells, but not neutrophils, to human umbilical vein EC monolayers. The increase in T cell adhesion induced by IL-4 was dose dependent (ED50 = 5 U/ml) and peaked around 33 U/ml. No increase in adhesion of neutrophils was observed at concentrations of IL-4 up to 1000 U/ml. The kinetic of the increase in T cell adhesion exhibited a steady rise peaking between 18 and 24 h before returning to basal levels by 72 h. The IL-4 specificity of the effect was confirmed by the ability of neutralizing anti-IL-4, but not anti-TNF, antibodies to abolish the effect. The increase in T cell-EC adhesion was due to an effect of IL-4 on EC inasmuch as preincubation of the T cells with IL-4 did not increase T cell binding. Furthermore, preincubation of A549 epithelial cell line monolayers with IL-4 caused no increase in T cell binding whereas A549 cells and EC showed a similarly enhanced adhesiveness for T cells after preincubation with IL-1, TNF, or IFN-gamma. EC treated with IL-4 retained their increased adhesiveness for T cells after light fixation, suggesting that IL-4 up-regulates binding by increasing the expression or accessibility of EC surface receptors for lymphocytes. Although antibodies to intercellular adhesion molecule-1 (CD54) and the beta-chain (CD18) of lymphocyte function-associated Ag-1 (CD11a/CD18) partially inhibited T cell adhesion to unstimulated EC, they did not affect the increase in adhesion due to IL-4 stimulation, indicating that the increased binding resulted from the generation of an alternative binding receptor(s) on the EC membrane. These findings suggest that IL-4 may play a role in the selective recruitment of T cells into sites of immune-mediated chronic inflammation.