Apparent Maintenance Research Articles

Neurogenic modulation and vasoactive peptides in microvascular biology

Acute neurogenic inflammation is observed after topical application of the TrpV1 agonist capsaicin to the mouse ear. Inflammatory oedema is not detectable in the substance P NK1 receptor knockout mouse, indicating the importance of this receptor in mediating neurogenic oedema formation. However, neurogenic vasodilatation remains not only in the NK1 receptor knockout mouse but also in the CGRP knockout mouse after sensory nerve stimulation. Interestingly, it is abolished when both the CGRP and substance P dilator pathways are blocked. This has led us to suggest that there is a facilitatory interaction between endogenously released CGRP and substance P, such that when both are present there is redundancy in terms of dilator mechanisms. This could be relevant to the apparent maintenance of peripheral vasodilator tone, when either NK1 or CGRP receptor antagonists are given systemically for evaluation as novel therapeutic agents. The relative activity of CGRP and the structurally related non‐neuropeptide adrenomedullin have been compared. Adrenomedullin is less potent as a vasodilator, but functionally relevant concentrations can be induced in inflamed skin. Both peptides, as a consequence of their vasodilator activity, influence inflammatory processes (e.g. oedema and cell accumulation) acting through the CGRP receptor. Thus, adrenomedullin can act as a non‐neuronally derived CGRP agonist in skin.

The Effects of Selection for early (day) and late (dusk) Mating lines of Hybrids ofBactrocera tryoniandBactrocera neohumeralis

Bactrocera neohumeralis and Bactrocera tryoni are closely related tephritid fruit fly species. B. neohumeralis mates throughout the day (in bright light) and B. tryoni mates at dusk. The two species can also be distinguished by the colour of their calli (prothoracic sclerites) which are brown and yellow, respectively. The F1 hybrids can mate both in bright light just before dusk and during dusk and have calli that are partly brown and partly yellow. The F2 hybrids have a wider range of callus patterns and mating occurs more widely in the day as well as at dusk. We directly selected hybrid stocks for mating time, creating 'early' (day-mating) and 'late' (dusk-mating) lines. As an apparently inadvertent consequence, the two types of line respectively had predominantly brown and predominantly yellow calli and thus came to closely resemble the original two species in both behaviour and appearance. Lines that were evenly selected (half for day and half for dusk) essentially retained the mating pattern of F2 hybrids. Selection for callus colour alone also affected the distribution of mating times in a predictable way. We propose a genetical model to account for the results and discuss them in the light of the apparent maintenance of species integrity in nature.

False autoregulation (pseudoautoregulation) in patients with severe head injury. Its importance in CPP management.

False autoregulation has been described as an alteration of autoregulation in which the apparent maintenance of a constant cerebral blood flow (CBF) when increasing cerebral perfusion pressure (CPP) is due to an increase in brain tissue pressure. The objective of our study was to investigate how often false autoregulation occurred in patients with a severe head injury. In forty-six patients with a moderate or severe head injury autoregulation was studied using arteriojugular differences of oxygen (AVDO2) to estimate changes in CBF after inducing arterial hypertension with phenylephrine. Changes in mean arterial blood pressure (MABP), intracranial pressure (ICP), cerebral perfusion pressure (CPP) and AVDO2 were calculated before and after inducing hypertension. Ninety-five episodes of provoked hypertension were studied in 46 patients. In 28 tests (29.5%) a constant or even reduced CBF was detected simultaneously with a median increase in parenchymal ICP of 8.5 mm Hg (false autoregulation). In this group the median of the induced increase in MABP was 20.6 mm Hg with a median increase in CPP of 11.5 mm Hg. From our data we can conclude that false autoregulation is frequently found in patients after a severe head injury. Increasing MABP to obtain a better CPP in these patients is not beneficial because CBF is not modified or may even be reduced.

Whole-body urea cycling and protein turnover during hyperphagia and dormancy in growing bears (Ursus americanusandU.arctos)

Subadult bears were studied during their autumn hyperphagia (n = 3) and winter dormancy (n = 6). Urea kinetics were measured with14C- and15N-urea, protein turnover was estimated with15N-glycine, and body composition was assessed with3H-water. Reduced amino acid degradation in winter was indicated by declines in plasma urea and aminotransferase activities, and lower urea production than in autumn (4.7 vs. 27.5 mmol urea-N∙kg−0.75∙d−1). Only 7.5% of urea produced in hyperphagic bears was degraded and just 1.1% of the degraded N reutilized as amino-N. Dormant bears reutilized 99.7% of urea produced, indicating thorough microbial ureolysis and urea-N resorption. Low rates of body N loss during dormancy suggested losses of non-urea N as creatinine. Protein turnover rates (15.2–21.5 g∙kg−0.75∙d−1) were similar between seasons and reflected the apparent maintenance of hepatic, intestinal, and muscular functions through dormancy. Protein synthesis accounted for 32% of energy expended in dormancy, which was mainly (91.5%) derived from fat oxidation. Consistent organ function and body temperature in dormant bears enables recycling of urea-N, which minimizes body protein loss and conserves mobility. In comparison with heterothermic hibernation, ursid dormancy would provide greater flexibility during winter and facilitate rapid resumption of foraging and growth in spring.

Characterization of anther culture-derived cell suspensions exclusively regenerating green plantlets in wheat (Triticum aestivum L.)

A reproducible procedure for deriving highly regenerable cell suspensions that can readily and consistently regenerate green plantlets in wheat is described. Initiation and selection of the right type of callus from anther cultures, which consisted of friable early embryogenic portions that can easily disperse in liquid medium was important for the establishment of rapidly growing embryogenic suspensions. Using this type of inoculum no significant variation between three different independent replications was noted when cell suspensions from eleven specially recombined doubled haploid lines were maintained on General medium supplemented with dicamba and a predominance of amino acid nitrogen. This approach also enhanced a long-term embryogenic competence of the cell cultures, with some of the suspensions retaining their morphogenic capacity over a period of more than 15 months. Depending on the medium composition high frequencies of embryogenesis (over 70%) and green plantlet regeneration (repeatedly producing 90–100% of green regenerants) were obtained from the cell aggregates for most of the embryogenic cell lines. Potential advantages of anther culture-derived embryogenic cell suspensions for transformation purposes are the high number of cell lines which can be established routinely and the apparent maintenance of a stable haploid genome by the regenerants in culture. It is anticipated that an increased use of anther or microspore derived doubled haploid techniques in future wheat breeding programmes may favour selection in the breeding material of plant types generally responsive to such protocols.

Pasteurella haemolytica Lipopolysaccharide-induced Cytotoxicity in Bovine Pulmonary Artery Endothelial Monolayers: Inhibition by Indomethacin

Exposure of bovine pulmonary artery endothelial cells to Pasteurella haemolytica lipopolysaccharide caused severe morphologic changes. Initially, there was dilatation of the rough endoplasmic reticulum and mitochondrial swelling followed by cell retraction, membrane bleb formation, and cell detachment. The affected endothelial cells had severe membrane damage resulting in the leakage of lactate dehydrogenase. Indomethacin in concentrations of 0.5 mM or greater caused marked decreases in the lipopolysaccharide-induced leakage of lactate dehydrogenase. Indomethacin at 5 mM also caused a marked reduction of the lipopolysaccharide-induced morphologic changes resulting in apparent maintenance of the monolayer integrity for 8 hours versus 1 hour in the lipopolysaccharide-treated control. A marked decrease in the cell and nuclear membrane effects resulted, but the rough endoplasmic reticulum dilatation and mitochondrial changes proceeded. These results indicate that indomethacin does not prevent lipopolysaccharide binding but interferes with later events in lipopolysaccharide-induced cytotoxicity in the bovine pulmonary endothelial cell. The concentration of indomethacin required to produce this inhibition suggests that the primary mechanism is not cyclooxygenase inhibition.

Energetics of arginine and lysine transport by whole cells and membrane vesicles of strain SR, a monensin-sensitive ruminal bacterium.

Strain SR, a monensin-sensitive, ammonia-producing ruminal bacterium, grew rapidly on arginine and lysine, but only if sodium was present. Arginine transport could be driven by either an electrical potential or a chemical gradient of sodium. Arginine was converted to ornithine, and it appeared that ornithine efflux created a sodium gradient which in turn drove arginine transport. There was a linear decline in arginine transport as pH was decreased from 7.5 to 5.5, and the cells did not grow at a pH less than 6.0. The Eadie-Hofstee plot was biphasic, and arginine could also be taken by a high-capacity diffusion mechanism. Because arginine was a strong inhibitor of lysine transport and lysine was a weak inhibitor of arginine transport, it appeared that both lysine and arginine were taken up by an arginine-lysine carrier which had a preference for arginine. The rate of lysine fermentation was always proportional to the extracellular lysine concentration, and facilitated diffusion was the dominant mechanism of lysine transport. When SR was grown in continuous culture on arginine or lysine, the theoretical maximal growth yield was similar (13 g of cells per mol of ATP), but the apparent maintenance energy requirement for arginine was greater than lysine (9.4 versus 4.4 mmol of ATP per g of cells per h). On the basis of differences in yield and maintenance energy, it appeared that active arginine transport accounted for approximately 40% of the total ATP.

Kinetic analysis of the growth of Chlorella vulgaris

The energy of the total transmitted light was subtracted from that of the incident light in a culture vessel and the difference was divided by the weight of cells. The value thus obtained was defined as the amount, E(x), of light energy absorbed per unit cell weight per unit time.Batch and continuous cultures of Chlorella vulgaris were carried out at 30 degrees C in the pH range of 6.4-6.7 while restricting illumination. Next the specific growth rate, mu, in the batch culture and the fixed dilution rate, D, in the continuous culture were plotted against E(x). The results showed that the relation between D and E(x) can be expressed in a Michaelis-Menten equation, where the maximal specific growth rate is 0.24 h (-1) and the saturation constant is 6.58 kcal/g . h.Cell concentration calculated by substituting the apparent concentration, X(e), of incubated cells and the apparent maintenance constant, M(e), for this equation agreed with that observed in almost all growth phases. Furthermore, from the change of chlorophyll productivity and the relationship between D and E(x) expressed in this equation, it is assumed that E(x) involves the light energy directly utilized in photosynthesis in the cells and that which is converted into, e.g., heat. This equation also indicated that a maximum in the growth yield existed. Then the growth yield of 0.029 g/kcal obtained at the incident light of 1.46 or 2.63 cal/cm(2) . h was maximum (maximal conversion efficiency of light energy, 15.6%).These results indicate that this method of deriving the equation for the growth rate from this study is a useful procedure for obtaining bioengineering findings.

Chloroplast Osmotic Adjustment and Water Stress Effects on Photosynthesis

Previous studies have suggested that chloroplast stromal volume reduction may mediate the inhibition of photosynthesis under water stress. In this study, the effects of spinach (Spinacia oleracea, var ;Winter Bloomsdale') plant water deficits on chloroplast photosynthetic capacity, solute concentrations in chloroplasts, and chloroplast volume were studied. In situ (gas exchange) and in vitro measurements indicated that chloroplast photosynthetic capacity was maintained during initial leaf water potential (Psi(w)) and relative water content (RWC) decline. During the latter part of the stress period, photosynthesis dropped precipitously. Chloroplast stromal volume apparently remained constant during the initial period of decline in RWC, but as leaf Psi(w) reached -1.2 megapascals, stromal volume began to decline. The apparent maintenance of stromal volume over the initial RWC decline during a stress cycle suggested that chloroplasts are capable of osmotic adjustment in response to leaf water deficits. This hypothesis was confirmed by measuring chloroplast solute levels, which increased during stress. The results of these experiments suggest that stromal volume reduction in situ may be associated with loss of photosynthetic capacity and that one mechanism of photosynthetic acclimation to low Psi(w) may involve stromal volume maintenance.

Behavioral treatment of schizophrenic delusions: A single-case experimental analysis

A 22-year-old white male who met the criteria for schizophrenia, paranoid, chronic (DSM-III 295.32) was treated via a social reinforcement-assisted cognitive intervention. Treatment sessions were designed to directly modify “strength of belief” in the validity of specific delusional ideas utilizing a collaborative style along with active avoidance of confrontation. An ABAB experimental design was used to allow assessment of changes in strength of belief as a function of presence or absence of the treatment intervention. During treatment phases, therapy sessions along with completion of a “self-therapy” log focused on repeated evaluation and interpretation of delusional beliefs. Social reinforcement was used to enhance compliance. Baseline phases included placebo control sessions. Dependent measures which showed improvement during treatment phases of the experiment were self-reports of (1) frequency and of (2) strength of belief in the validity of schizophrenic delusions, and (3) frequency of need for administration of p.r.n. medication (thorazine 100 mg) given by nursing staff blind to the experimental conditions. A three-month follow-up showed partial persistence of therapeutic gains, including apparent maintenance of acquired “metacognitive skills.” Replication studies incorporating specific treatment components are suggested.

Stimulation by epinephrine of histamine synthesis by rat peritoneal fluid mast cells in vitro

In vitro exposure of mast but not of other cells of rat peritoneal fluid to epinephrine leads, within 1 min, to progressing levels of histamine in both fluid and sedimentable phases of the incubates, which present no increase in their free/total histamine ratio. Histamine increase was blocked by α-fluoromethyl histidine (αFMH), acting after a significant lag period. When compared with controls under the electron microscope, epinephrine-treated mast cells show less electron-dense, swollen intercellular granules, apparent maintenance of cell membrane continuity and an apparent decrease of peripheral finger-like projections. Histamine accumulation by epinephrine-treated mast cells may be the result of an enhanced ability of pre-formed mast cell histidine decarboxylase to attack its cell-borne substrate, consequent to an unfolding of the cell membrane during cell tumefaction evoked by epinephrine.

Energy partitioning and growth in mice selected for high and low body weight.

SUMMARYThe partitioning of digestible energy intake on anad libitumdiet of standard mouse nuts was investigated in mice selected for high and low body weight at eight weeks and in an unselected control population. In selected mice aged from four to six weeks and housed at a temperature of 24·5 °C, almost all their energy intake could be attributed to basic maintenance and the deposition of extra protein and fat. Control mice, however, had an energy intake considerably in excess of their apparent maintenance and growth requirements. It was concluded that the unaccountable energy loss of the control line could be used to increase growth efficiency in the selected lines. The result is analogous to those obtained from studies on normal and obese mouse genotypes and indicates genetic changes in mechanisms controlling the conversion of food energy to heat.Provision of a nest to reduce thermoregulatory heat production caused a minor reduction in energy intake and a corresponding decrease in the energy discrepancy. There was no effect on growth.

A detailed evaluation of oral phosphate therapy in selected patients with primary hyperparathyroidism.

Recent studies have emphasized the pathophysiological importance of circulating 1,25-dihydroxyvitamin D ((1,25-(OH)2D] in the pathogenesis of hypercalciuria and renal stone formation in primary hyperparathyroidism. Reasoning that phosphate administration might be capable of reducing the plasma concentration of 1,25-(OH)2D in patients with a prominent 1,25-(OH)2D-mediated absorptive component to their disease, 10 carefully selected patients were treated with oral phosphate (1500 mg elemental phosphorus daily) for 1 yr. Phosphate treatment significantly reduced circulating 1,25-(OH)2D levels (84 to 56 pg/ml), the calciuric response to an oral calcium tolerance test (0.30 to 0.21 delta mg calcium/dl GF), and calcium excretion on an unrestricted calcium diet (438-269 mg/day), in essence reversing the absorptive pattern of abnormalities observed before treatment. This response, however, was accompanied by an increase in biochemical hyperparathyroidism, as assessed by circulating immunoreactive PTH and nephrogenous cAMP excretion. In patients with biochemical evidence of an increase in bone resorption before therapy, histomorphometric, radiographic, and biochemical data revealed a trend toward a reduction in bone turnover during phosphorus therapy, with an apparent maintenance of coupled bone resorption and bone formation. This trend, however, was of marginal statistical significance in the patient group as a whole. It is concluded 1) that phosphate therapy represents a viable medical alternative in selected patients with primary hyperparathyroidism, 2) that the net response in treated patients is multifaceted and complex, and 3) that the efficacy of phosphate therapy will ultimately depend upon its long term effects on skeletal homeostasis.

Maintenance Energy Requirements during Lactation in Rats

Evidence is presented to support the postulates that increases in apparent maintenance requirements during pregnancy and lactation are due, in part, to increases in relative weights and metabolic activities of vital organs. Rates of glucose, palmitate and pyruvate oxidation per unit weight in vitro were one-and-a-half to three times greater in liver, heart and intestine samples from pregnant and lactating as compared to nonlactating rats. Relative weights of liver, intestinal tract and heart were greater in lactating as compared to nonlactating rats. Weights of liver, heart and intestines are functions of feed intake and may parallel a previously reported logistic function which relates feed intake to apparent maintenance requirements. A theoretical calculation indicated that an increase in maintenance requirements from 100 kcal metabolizable energy/kilograms0.75 to 124 kcal metabolizable energy/kilograms0.75 during lactation in rats could be explained on the basis of changes in the relative weights of liver, intestines and heart.tissue weights maintenance pregnancy lactation

Very slow growth of Bacillus polymyxa: Stringent response and maintenance energy

Bacillus polymyxa grown in a recycling fermentor shows the same behavior previously observed with Escherichia coli: 3 successive growth phases. In the last 2 phases the growth rate is linear and the apparent maintenance energy demand rate and the molar growth yield are both independent of the specific growth rate, μ, and of the cells mass. The final phase of very slow growth is an indefinitely prolonged state of strong, stringent control, the regulatory system based on guanosine 3′-diphosphate 5′-diphosphate, and guanosine 3′-diphosphate 5′-triphosphate. The maximum cost of this stringent response is calculated to be 9% of the energy available to these energy-limited cells. There is a further energy cost contained in substantial amounts of DNA, RNA, and protein released from the cells during the latter 2 growth phases. The cost of production of these extra cellular anabolites ranges from 8–11% of the available energy.

Manipulating metabolic parameters to improve growth rate and milk secretion.

Several opportunities for improving animal efficiency through manipulation of metabolism are discussed. The first opportunity is through identification and selection of animals achieving close to theoretical efficiencies. Based upon differences between highly efficient and average animals, the estimated opportunity for improvement is 20%. A second opportunity for improvement is through manipulation of apparent maintenance requirements. Several contributors to differences in efficiencies are considered. One is the contribution of differences in relative organ weights to differences in apparent maintenance requirements. A potential benefit in the order of 10 to 20% through selection or manipulation seems possible. Manipulations of ion transport and protein turnover could yield maximum benefits of 30 and 15%, respectively. However, complete elimination of these processes is not feasible. Without ion transport, membrane potentials would not be maintained and, without turnover, many important regulatory processes would be affected. the limit to manipulation of these characteristics is unknown. A third opportunity for improvement of animal efficiency is through improvement of apparent biosynthetic efficiency by manipulation of patterns of nutrient utilization. If we could produce, through hormonal or other types of manipulations, an optimum pattern of nutrient use, decreases in heat increments of production in growing animals in the order of 50% might be achieved.

Apparent maintenance of cytochrome P 450 by nicotinamide in primary cultures of rat hepatocytes

The sole addition of a high, unphysiological, concentration of nicotinamide (25 mM) to a cell culture medium was found to maintain the cytochrome P 450 concentration of rat hepatocytes cultured for 24 hours at 71% of the level found in intact liver, whilst hepatocytes cultured without nicotinamide contained only 20% of their initial cytochrome P 450. Furthermore the P 450 concentration of hepatocytes cultured for 24 hours in the presence of 25 mM nicotinamide could be increased to the same level as found in intact rat liver by the inclusion of 1 mM nicotinamide into the medium used for cell isolation. Although the mechanism of action of nicotinamide is unknown this simple system for the maintenance of cytochrome P 450 in hepatocyte culture could provide the opportunity to study, under defined conditions in vitro , the factors that regulate cytochrome P 450 and hence determine hepatotoxicity and hepatocarcinogenesis.

AXON SPROUTING INTO SEGMENTS OF RAT SPINAL CORD ADJACENT TO THE SITE OF A PREVIOUS TRANSECTION

Transection of the spinal cord is adult Long-Evans Hooded rats is followed by the formation of a connective tissue matrix in the lesion site and the rapid erosion of the neural elements above and below this zone particularly within the dorsal white columns. In the period between 15--45 days after operation two significant events begin to occur. First the injured surfaces of the divided cord become invested by a glial limiting membrane (glia limitans) and, concomitantly, large numbers of axons ensheathed by Schwann cells sprout into the scar matrix and along the eroded dorsal column region. The injured surface of the spinal cord is highly irregular with deep, collagen-filled rifts into which the sprouting axons may probe and penetrate into the adjacent normal neuropil. Electron microscopic examination generally reveals the interposed glia limitans and that these fibres are usually restricted to the peripheral environment. However, as some axons approach the reconstituting glia limitans, they are enveloped by an astrocytic cytoplasmic process which may either displace the Schwann cell or encompass it together with the enclosed axons. This last phenomenon appears to precede the entry of some axons into the neuropil and suggests that the glia limitans may not necessarily represent an impenetrable barrier to the passage of regenerating axons into the CNS. Apparent maintenance of most of these fibres for periods of up to 3 months may suggest that viable, functional synapses were established upon available neuronal elements, but clear evidence of this could not be documented.

The distribution of blood rheological parameters in the microvasculature of cat mesentery.

In vivo studies of the rheological behavior of blood in the microcirculation were conducted by direct in situ measurements in cat mesentery. Upstream to downstream pressure drops were measured in unbranched arterioles, capillaries, and venules, with diameters from 7 to 58 micrometer. Simultaneous measurements of red cell velocity and vessel geometry facilitated computation of bulk velocity, pressure gradient, apparent viscosity, wall shear stress, and resistance. Arteriovenous distributions of these parameters revealed the following. Maximum pressure gradient (0.015 cm H20/micrometer) occurs in the true capillaries (7 micrometer in diameter); intravascular wall shear stress averaged 47.1 dynes/cm2 in arterioles and 29.0 dynes/cm2 in venules. Extreme values as great as 200 dynes/cm2 were observed in a few shunting arterioles. Apparent viscosity averaged 3.59 cP in arterioles, 5.15 cP in venules, and 4.22 cP overall. Intravascular resistance per unit length of microvessel varied with luminal diameter as a power law function with exponents of -4.04 for arterioles, -3.94 for venules, and -3.99 for all vessels combined. This apparent maintenance of Poiseuille's law is attributed to the opposing processes of hematocrit reduction and decreasing shear rate as blood is dispersed in successive arteriolar segments, and the converse action of these processes in the venous confluences which lessen the extent of network variations in apparent viscosity. Reductions in bulk velocity from the normal flow state to below 0.5 mm/sec resulted in increases in apparent viscosity by a factor of 2 to 10, which are attributed primarily to obstruction of the lumen by leukocyte-endothelium adhesion.

Interaction between the plasmid R6K andEscherichia coliwith defective DNA polymerase I

SUMMARYInitial experiments demonstrated that the plasmid R6K cannot be transferred to or maintained readily in theE. coliDNA polymerase I deficient strain JG138polA1. Results withE. coliMM386polA12(R6K), which has a temperature sensitive polymerase I enzyme, showed cell division becomes abnormal when the polymerase I enzyme of the host bacteria is inactivated at the restrictive temperature. Under conditions of polymerase I deficiency, R6K replication, as measured by monitoring R-factor-mediated β-lactamase activity, also becomes abnormal with the loss of multiple R6K copies per cell and the apparent maintenance of a single R-factor copy per cell.