Age-related growth of the spleen in normal and glucocorticoid treated rats

1. Age-related changes in the growth, nucleic acid content and protein turnover of the spleen have been studied in normal male rats. 2. A rapid and marked atrophy of the spleen, was found 24 hr after exposure to cortisone or dexamethasone; increased rates of protein breakdown being primarily responsible. 3. Nonetheless, the total amount of protein synthesized in the spleen (measured in vivo) was significantly decreased (30-50%) 24 hr after exposure to these steroids. 4. This compared with only a 15% decrease in whole body protein synthesis, indicating a more pronounced hormonal effect on the spleen than on most other body tissues.     

Differential heating of trunk and extremities. Effects on thermoregulation mechanisms

Experiments in which the whole human body was heated or cooled are compared with others in which one extremity (arm or leg) was simultaneously cooled or heated. With a warm load on the rest of the body resulting in general sweating, a cold load on one extremity did not evoke local shivering; with general body cooling, heating one limb did not stop the shivering. Skin temperatures of the other parts of the body were not influenced by warming or cooling one extremity. Evaporative heat loss was influenced by local, mean skin and core temperature, whereas shivering did not depend on local temperature, and vasomotor control seemed to be controlled predominantly by central temperatures. A cold load on an extremity during whole body heating in most cases induced an oscillatory behaviour of core temperature and of the evaporative heat loss from the body and the extremity. It is assumed that local, mean skin and core temperatures influence the three autonomous effector systems to very different degree.     

Noncoordinate synthesis of the vitellogenin proteins in tissues of Drosophila grimshawi

In analyzing the in vitro pattern of protein synthesis by the fat body and ovaries of the Hawaiian species Drosophila grimshawi, we have found that the ovaries synthesize much more protein than the female fat body and that the majority of the synthesized proteins are retained by the ovarian tissues. In contrast, the fat body secrets most of the proteins into the culture medium. Vitellogenins are the major class of proteins synthesized and released into the medium by both tissues. The synthesis of the three vitellogenin proteins (V1, V2, V3) is noncoordinate in the two tissues. Ovaries synthesize much more of the V2 protein, less V1 and very little V3, whereas fat body synthesizes more V1 protein with lesser quantities of the other two. The follicle cells were identified as the site of ovarian vitellogenin synthesis in D. grimshawi, confirming the findings in D. melanogaster. In D. grimshawi, the three vitellogenins are synthesized by the follicle cells in a noncoordinate and developmentally regulated manner. V2 and V1 are the predominant proteins at the onset of vitellogenesis (S8-9); their production peaks together with that of V3 a few hours later (S10) and then decreases to quantities equal to that of V3 during early choriogenesis (S11). During active choriogenesis (S12), V2 and V1 cease to be synthesized, but V3 synthesis continues. The vitellogenins synthesized by the follicles in vitro are released into the medium and not incorporated into the oocyte.     

Muscle glutamine depletion in the intensive care unit

Glutamine is primarily synthesized in skeletal muscle and enables transfer of nitrogen to splanchnic tissues, kidneys and immune system. Discrepancy between increasing rates of glutamine utilization at whole body level and relative impairment of de novo synthesis in skeletal muscle leads to systemic glutamine deficiency and characterizes critical illness. Glutamine depletion at whole body level may contribute to gut, liver and immune system disfunctions, whereas its intramuscular deficiency may directly contribute to lean body mass loss. Severe intramuscular glutamine depletion also develops because of outward transport system upregulation, which is not counteracted by increased de novo synthesis. The negative impact of systemic glutamine depletion on critically ill patients is suggested both by the association between a lower plasma glutamine concentration and poor outcome and by a clear clinical benefit after glutamine supplementation. Enteral glutamine administration preferentially increases glutamine disposal in splanchnic tissues, whereas parenteral supplementation provides glutamine to the whole organism. Nonetheless, systemic administration was ineffective in preventing muscle depletion, due to a relative inability of skeletal muscle to seize glutamine from the bloodstream. Intramuscular glutamine depletion could be potentially counteracted by promoting de novo glutamine synthesis with pharmacological or nutritional interventions.     

Tissue-specific synthesis of yolk proteins in Caenorhabditis elegans

The primary site of yolk protein synthesis in the nematode, Caenorhabditis elegans, has been determined. In animals containing no gonadal cells (obtained by laser ablation of the gonadal precursor cells early in development), yolk proteins are present in abundance. This demonstrates that yolk proteins are made outside the gonad. An examination of proteins present in tissues isolated by dissection, and a comparison of proteins synthesized by isolated tissues incubated in vitro have identified the intestine as the major site of yolk protein synthesis. We propose that yolk proteins are synthesized in the intestine, secreted from the intestine into the body cavity, and taken up from the body cavity by the gonad to reach oocytes. The site of yolk protein synthesis has also been examined in four mutants that have largely male somatic tissues, but a hermaphrodite germ line. Here again, yolk proteins are produced by intestines in a hermaphrodite-specific manner. This suggests that sex determination is coordinately regulated in intestinal and germ line tissues.     

Monitoring of whole body cryotherapy effects by thermal imaging: preliminary report

In whole body cryotherapy the whole human body is exposed to low temperature below −100°C in a special room called cryogenic chamber for a very short period of time (2–3 minutes). The impact of cold can cause many different biochemical and physiological reactions of the organism.The skin temperature response due to whole body cryotherapy was studied by means of infrared measurements. The thermograms of chosen body parts of patients suffering from low back pain were performed before and after whole body cooling on the 1^st, 5^th and the last (10^th) day of medical treatment. Infrared imaging performed after cold impact owing to the enhancement of the skin temperature profile may reveal a slight decrease of the inflammatory states as a result of the 10 sessions of cryotherapy.     

The protective effect of thermal treatment before irradiation on CFUs from bone marrow of mice: a potential involvement of heat shock proteins

It was studied on mice how prior whole body hyperthemia affects a colony-forming ability of bone marrow after gamma-irradiation. It was found that heating of the animals (42 degrees C, 10 min) 18-22 h before their total irradiation (4 Gy) increases 2-fold the level of CFUs8 and CFUs12 determined in the spleen exotest. The induced radioresistance correlated with accumulation of heat shock proteins, HSP70 and HSP25, in tissues of preheated mice. Injection of quercetin (a selective inhibitor of the heat shock protein synthesis) 0.5 h before the heating fully abolished both the subsequent heat shock protein accumulation and the rise in CFUs populations as compared with control. It is suggested that heat shock proteins, whose expression increases in response to hyperthermia, can play a role of endogenous radioprotectors. Possible mechanisms of their protective action under irradiation are discussed.     

Changes in tissue basophils of skin and lymphoid organs in total, deep hypothermia

The experiments have been performed on 60 white female rats with body mass 180-200 g. Under ether anesthesia the animals are cooled up to +18 degrees C of the rectal temperature, and then warmed up to +37 degrees C. Using certain morphometrical methods at staining paraffin slices with toluidine++ blue, tissue basophils (TB) in the dermis++, hypoderma, lymph nodes, thymus and spleen are studied. In response to cooling degranulation increases, some part of TB in all the organs is destroyed, this results in decreasing amount of the cells (except the thymus, where TB amount increases). The greatest pronounced decrease of TB amount occurs in the lymph node. The restorative reaction during the post-hypothermic period depends on the initial lesion. There are no TB in the spleen. In different organs TB demonstrate various sensitivity to hypothermia, that is evidently connected with their microenvironment and initial state at the moment of the experiment, as well as with various role of the organs investigated in the defensive-adaptive reactions of the organism.     

Acute stimulation of steroidogenesis in corpus luteum and adrenal cortex by peptide hormones. Rapid induction of a similar protein in both tissues

Two-dimensional electrophoresis was used to detect a protein (ic) synthesized in rat corpus luteum cells in response to acute stimulation by human chorionic gonadotropin or dibutyryl cyclic AMP. This induced protein ic is isoelectric at pH 6.5 (isoelectric focusing) and has an apparent molecular weight of 28,000 (sodium dodecyl sulfate electrophoresis). The human chorionic gonadotropin or dibutyryl cyclic AMP dose response and time course of synthesis of the protein parallel those of progesterone synthesis in stimulated luteal cells. Additionally, cycloheximide, which inhibits the increase in progesterone formation caused by human chorionic gonadotropin or cAMP, also inhibits the synthesis of ic. Proteolytic polypeptide mapping suggests that ic has a very similar primary structure to another protein (pc), which has the same molecular weight as ic, differs from ic in pI, and is synthesized only in unstimulated cells. These polypeptide maps also demonstrate the close similarity of pc and ic to two proteins p and i, synthesized in control and in adrenocorticotropic hormone-stimulated rat adrenal cortex cells, respectively (Krueger, R. J. and Orme-Johnson, N. R. (1983) J. Biol. Chem. 258, 10159-10167). In both adrenal cortex and corpus luteum, binding of a tissue-specific polypeptide hormone acts via cAMP to cause increased steroidogenesis and induction of the synthesis of protein i (ic), with the same time course and hormone dose dependence. Also in both tissues, inhibition of protein synthesis at the level of translation (e.g. by cycloheximide addition) causes inhibition of i (ic) synthesis and of stimulated steroid production. This close correlation between the two different tissues in conditions which cause induction of the synthesis of these proteins suggests that the proteins may be common intermediaries in the control by polypeptide hormones of steroidogenesis in endocrine tissues.     

Protein synthesis in heterotopically transplanted rat hearts

This study examines protein synthesis in heterotopically transplanted rat hearts and several tissues of recipient rats. Donor hearts and recipient tissues synthesized many of the normally occurring proteins observed in tissues of unstressed rats. In addition, a stress-induced protein with a molecular mass of 71 kilodaltons was synthesized in donor heart, recipient heart and lung. Donor hearts incorporated more L-[35S]-methionine than did recipient hearts. Tissues of recipient rats also incorporated more label than the respective tissues of sham-recipient rats. These results suggest that ischemia, endured by the donor hearts during transplantation, induced these changes in protein synthesis.     

Protein turnover and growth in the whole body, liver and kidney of the rat from the foetus to senility.

Changes in the growth and protein turnover (measured in vivo) of the rat liver, kidney and whole body were studied between 16 days of life in utero and 105 weeks post partum. Tissue and whole-body growth were related to changes in both cellular hyperplasia (i.e. changes in DNA) and hypertrophy (protein/DNA values) and to the protein composition within the enlarging tissue mass. The suitability of using a single large dose of phenylalanine for measuring the rates of protein synthesis during both pre- and post-natal life was established. The declining growth rates in the whole animal and the two visceral tissues were then explained by developmental changes in the fractional rates of protein synthesis and breakdown, turnover rates being age-for-age higher in the liver than in the kidney, which in turn were higher than those measured in the whole animal. The declining fractional rates of synthesis in both tissues and the whole body with increasing age were related to changes in the tissues' ribosomal capacity and activity. The fall in the hepatic rate between 18 and 20 days of foetal life (from 134 to 98% per day) corresponded to a decrease in both the ribosomal capacity and the rate of synthesis per ribosome. No significant changes in any of these parameters were, however, found in the liver between weaning (3 weeks) and senility (105 weeks). In contrast, the fractional synthetic (and degradative) rates progressively declined in the kidney (from 95 to 24% per day) and whole body (from 70 to 11% per day) throughout both pre- and post-natal life, mainly as a consequence of a progressive decline in the ribosomal capacity, but with some fall in the ribosomal activity also during foetal life. The age-related contributions of these visceral tissues to the total amount of protein synthesized per day by the whole animal were determined. The renal contribution remained fairly constant at 1.6-2.9%, whereas the hepatic contribution declined from 56 to 11%, with increasing age. Approximate-steady-state conditions were reached at, and between, 44 and 105 weeks post partum, the half-life values of mixed whole-body, kidney and liver proteins being 6.4, 3.0 and 1.5 days, respectively, at 105 weeks.     

Cellular origin of prealbumin in the rat

The synthetic rate of prealbumin and albumin in primary monolayer cultures of rat hepatocytes was measured by immunochemical methods. The isolated hepatocytes synthesized these proteins in the same ratio as that previously found for the whole body synthesis in vivo. It is concluded that the hepatocytes synthesize the main part of prealbumin in the rat.     

PROTEIN AND NUCLEIC ACID METABOLISM IN INSECT FAT BODY

1. The appearance of larval fat body as seen under the light or electron microscope depends on the nutritional state of the larva and on the stage of larval development at which the fat body is observed. 2. Early in the last larval instar the cells usually possess a well-developed endo-plasmic reticulum rich in ribosomes, numerous mitochondria, glycogen granules, a Golgi complex and fat droplets, while later in the instar the endoplasmic reticulum is much reduced and mitochondria are few, but glycogen and fat droplets are present in greater amount together with the appearance of large numbers of proteinaceous spheres. 3. Early in the last instar the fat body synthesizes proteins and exports them into the blood, while later in the instar proteins are sequestered from the blood into the fat body. 4. The rate of protein synthesis by the fat body is high in the early to mid part of the last instar, but then falls off rapidly to a low level, at which it remains until the larva pupates. In diapausing pupae, protein synthesis remains at this low level. 5. The similarity between the electrophoretic patterns of proteins from the fat body and those from the blood provides strong evidence that the fat body is the site of synthesis of many of the blood proteins. 6. Some of the blood proteins have been shown to possess enzymic properties, while others are thought to play a role in the transportation of various types of compounds. 7. Ecdysone and juvenile hormone both stimulate the rate of protein synthesis by larval fat body. Protein synthesis in fat body from diapausing pupae is stimulated after injury to the pupae. 8. The appearance of adult fat body and the amount of protein it contains is often closely linked with the nutritional and reproductive states of the insect. 9. An important role of the fat body in the adult female insect is the synthesis of yolk proteins, which are released into the blood and then taken up by the developing oocytes. This synthesis and uptake are under the control of hormones secreted by the corpora allata and by the median neurosecretory cells of the pars intercerebralis. 10. The RNA content of fat body in final-instar larvae is not constant throughout the instar. In some larvae it is at its highest level early in the instar, falling to a low level as the instar progresses, while in other larvae (e.g. Calliphora) the level of RNA in fat body does not decrease as the instar progresses. 11. In some dipterous insects the base composition of total RNA is DNA-like in that the guanine + cytosine content is low, accounting for 40 % of the bases. A similar composition is seen in rapidly labelled RNA isolated from insects of other orders (Coleoptera and Lepidoptera), but the base content of total RNA from these latter insects resembles ribosomal RNA from vertebrate tissues in that it has a high (ca. 60 %) guanine + cytosine content. 12. The RNA/DNA ratios in blowfly larval tissues are high compared with those found in any vertebrate tissue. 13. In larval fat body, RNA synthesis is low at the time of a moult, increases during the early and mid-instar period and subsequently falls during the latter part of the instar. During the pupal period, especially during pupal diapause, the rate of RNA synthesis is very low and then increases during the subsequent development of the pharate adult. Injury to diapausing pupae results in an increased rate of RNA synthesis in most of their tissues. 14. Ecdysone and juvenile hormone both stimulate RNA and DNA synthesis in larval and adult fat body and in other tissues, although there is evidence that in some tissues these two hormones may act antagonistically to each other. The insecticide DDT also has been shown to stimulate RNA synthesis in tissues of adult insects.     

The heat-shock reaction is disturbed in a Drosophila virilis strain incapable of a neurohormonal stress reaction

Cellular stress response was investigated in two lines of D. virilis: wild type and line with disturbed neurohormonal stress-reaction. Analysis of proteins, synthesized in salivary glands of larvae of both lines under heat stress, revealed malfunction in heat shock reaction of mutant specimen. This malfunction expresses in decreased level of heat shock protein synthesis. Analysis of electrophoretic spectra of proteins from homogenates of imagoes of both lines maintained under normal conditions and those exposed to heat (38 degrees C, 60 min.) revealed correlation between protein spectrum and physiological state of the organism. Interlinear differences by proteins spectra in normal condition, controlled by a single gene (or by block of closely linked genes), were found. The question if there is a common genetic control for the neurohormonal stress-reaction and cellular stress response is discussed.     

Influence of cooling temperature and duration on cold adaptation of Lactobacillus acidophilus RD758

The effect of different cooling temperatures and durations on resistance to freezing and to frozen storage at -20 degrees C in Lactobacillus acidophilus RD758 was studied, by using a central composite rotatable design. A cold adaptation was observed when the cells were maintained at moderate temperature (26 degrees C) for a long time (8h) before being cooled to the final temperature of 15 degrees C. These conditions led to a low rate of loss in acidification activity during frozen storage (0.64 minday(-1)) and a high residual acidification activity after 180 days of frozen storage (1011 min). The experimental design allowed us to determine optimal cooling conditions, which were established at 28 degrees C during 8h. Adaptation to cold temperatures was related to an increase in the unsaturated to saturated fatty acid ratio and in the relative cycC19:0 fatty acid concentration. Moreover, an increased synthesis of four specific proteins was observed as an adaptive response to the optimal cooling conditions. They included the stress protein ATP-dependent ClpP and two cold induced proteins: pyruvate kinase and a putative glycoprotein endopeptidase.     

Effects of evaporative cooling on the regulation of body water and milk production in crossbred Holstein cattle in a tropical environment

The aim of this study was to determine how evaporative cooling modifies body function with respect to water metabolism and other variables relevant to milk synthesis in crossbred cattle. The study was conducted on two groups of 0.875HF:0.125RS crossbred Holstein cattle (87.5%) housed in an open-sided barn with a tiled roof (non-cooled animals) and in a close-sided barn under an evaporative cooling system (cooled animals). The maximum ambient temperature and relative humidity for the non-cooled group were 33 degrees C and 61%, with the corresponding values for the evaporatively cooled barn being 28 degrees C and 84%, respectively. The temperature humidity index (THI) of under non-cooled conditions was higher (P < 0.05) than that in the cooled barn. Rectal temperatures and respiration rates of non-cooled animals were higher (P < 0.05) than those of cooled animals. Daily dry matter intake (DMI) of cooled animals was higher while water intakes were lower (P < 0.05) than those of non-cooled animals. The mean absolute values of plasma volume, blood volume, and extracellular fluid (ECF) of cooled animals were significantly higher (P < 0.05) than those of non-cooled animals throughout all stages of lactation. Milk yields of cooled animals were higher by 42%, 36% and 79% on average than those of non-cooled animals during early-, mid- and late-lactation, respectively. The decline in milk yields as lactation advances was markedly apparent in late-lactating non-cooled animals, while no significant changes in milk composition at different stages of lactation were observed in either group. Mean arterial plasma concentrations, arteriovenous concentration differences (A-V differences) and the extraction ratio across the mammary gland for acetate, glucose and triglyceride of cooled animals were not significantly different compared with values for non-cooled animals. No differences were seen in plasma hormonal levels for triiodotyronine (T(3)) and insulin-like growth factor-1 (IGF-1), but plasma cortisol and thyroxine (T(4)) levels tended to be lower in non-cooled animals. This study suggests that low cooling temperature accompanied by high humidity influences a galactopoietic effect, in part through increases in ECF, blood volume and plasma volume in association with an increase in DMI, which partitions the distribution of nutrients to the mammary gland for milk synthesis. Cooled animals were unable to maintain high milk yield as lactation advances even though a high level of body fluids was maintained during long-term cooled exposure. The decline in milk yield, coinciding with a decrease in net energy for lactation as lactation advances, could be attributed to a local change within the mammary gland.     

Cooling different body surfaces during upper and lower body exercise

The effect of varying the body surface area being cooled by a liquid microclimate system was evaluated during exercise heat-stress conditions. Six male subjects performed a total of six exercise (O2 uptake = 1.2 l/min) tests in a hot environment (ambient temperature = 38 degrees C, relative humidity = 30%) while dressed in clothing having low moisture permeability and high insulation. Each subject completed two upper body exercise (U; arm crank) tests: 1) with only the torso surface (T) cooled; and 2) with the surfaces of both the torso and upper arms (TA) cooled [coolant temperature at the inlet (Ti) was 20 degrees C for all upper body tests]. Each subject also completed four lower body exercise (L; walking) tests: 1) with only the T cooled (Ti = 20 degrees C); 2) with only the T cooled (Ti = 26 degrees C); 3) with torso, upper arm, and thigh surface (TAT) cooled (Ti = 20 degrees C); and 4) with TAT cooled (Ti = 26 degrees C). During U exercise, TA cooling had no effects compared with cooling only T. During L exercise, sweat rates, heart rates, and rectal temperature (Tre) changes were less with TAT cooling compared with cooling only the T. Altering Ti had no effect on Tre changes, but higher heart rates were observed with 26 than with 20 degrees C. These data indicate that cooling arms during upper body exercise provides no thermoregulatory advantage, although cooling the thigh surfaces during lower body exercise does provide an advantage.     

Induction of rapid cold hardening by cooling at ecologically relevant rates in Drosophila melanogaster

Over a decade ago it was hypothesized that the rapid cold hardening process allows an organism's overall cold tolerance to track changes in environmental temperature, as would occur in nature during diurnal thermal cycles. Although a number of studies have since focused on characterizing the rapid cold hardening process and on elucidating the physiological mechanisms upon which it is based, the ecological relevance of this phenomenon has received little attention. We present evidence that in Drosophila melanogaster rapid cold hardening can be induced during cooling at rates which occur naturally, and that the protection afforded in such a manner benefits the organism at ecologically relevant temperatures. Drosophila melanogaster cooled at natural rates (0.05 and 0.1 degrees C min(-1)) exhibited significantly higher survival after one hour of exposure to -7 and -8 degrees C than did those directly transferred to these temperatures or those cooled at 0.5, or 1.0 degrees C min(-1). Protection accrued throughout the cooling process (e.g., flies cooled to 0 degrees C were more cold tolerant than those cooled to 11 degrees C). Whereas D. melanogaster cooled at 1.0 degrees C min(-1) had a critical thermal minimum (i.e., the temperature at which torpor occurred) of 6.5+/-0.6 degrees C, those cooled at an ecologically relevant rate of 0.1 degrees C min(-1) had a significantly lower value of 3.9+/-0.9 degrees C.