Rapid increase in poly(A) (+) mRNA content following inhibition of protein synthesis

When resting 3T6 cells undergo a serum-induced transition to the growing state, the cytoplasmic content of ribosomal, transfer and messenger RNA increase as the cells prepare for DNA synthesis. The normal linear increase in mRNA content occurs even when the production of ribosomes is blocked. In this paper we determine the effect of inhibiting protein synthesis on the increase in poly(A) (+) mRNA content. Resting cells were serum stimulated in the presence of cycloheximide or puromycin at levels which inhibit protein synthesis by greater than 95%. Cytoplasmic poly(A) (+) mRNA content was determined at various times thereafter. We found that mRNA content increased five to ten times more rapidly in drug treated cells than in control cells stimulated in the absence of inhibitors. mRNA content increased 50–70% by one hour, and 60–90% by two hours following stimulation in the presence of inhibitor, and remained more or less constant thereafter. In contrast, mRNA content increased linearly in control stimulated cultures and did not double until about 15 hours after stimulation. The rapid increase in mRNA content is most likely the result of inhibition of protein synthesis rather than a secondary effect of the drug since the same observations were made in growth stimulated cells if protein synthesis was blocked with either puromycin or cycloheximide. A similar effect was also observed with resting 3T6, exponentially growing 3T6 and growing HeLa cells following exposure to cycloheximide, although the magnitude of the increase was less than that observed with growth stimulated cells. Puromycin had negligible effect on mRNA content in resting or exponentially growing cells. The rapid increase in cytoplasmic poly(A) (+) mRNA content was not due to rapid unbalanced export of nuclear poly(A) (+) RNA into the cytoplasm since there was no decrease in nuclear poly(A) content following serum stimulation in the presence of cycloheximide.     

Rapid increase in southern elephant seal genetic diversity after a founder event

Genetic diversity provides the raw material for populations to respond to changing environmental conditions. The evolution of diversity within populations is based on the accumulation of mutations and their retention or loss through selection and genetic drift, while migration can also introduce new variation. However, the extent to which population growth and sustained large population size can lead to rapid and significant increases in diversity has not been widely investigated. Here, we assess this empirically by applying approximate Bayesian computation to a novel ancient DNA dataset that spans the life of a southern elephant seal (Mirounga leonina) population, from initial founding approximately 7000 years ago to eventual extinction within the past millennium. We find that rapid population growth and sustained large population size can explain substantial increases in population genetic diversity over a period of several hundred generations, subsequently lost when the population went to extinction. Results suggest that the impact of diversity introduced through migration was relatively minor. We thus demonstrate, by examining genetic diversity across the life of a population, that environmental change could generate the raw material for adaptive evolution over a very short evolutionary time scale through rapid establishment of a large, stable population.     

Rapid senescence‐like response after acute injury

Cellular senescence is a state of irreversible growth arrest. Short‐term programmed senescence such as in embryonic development and slowly progressing senescence as in aging are both well described. However, acute senescence in living organisms is not well understood. We hypothesized that hemorrhagic shock injury (HI) caused by whole body hypoxia and nutrient deprivation, resulting in organ dysfunction due to severe blood loss, could lead to acute senescence in vivo. Our experiments, for the first time, demonstrate a rapidly emerged, senolytics‐responsive, senescence‐like response in the rat liver in less than five hr following hemorrhagic shock. We conclude that the senescence, or pseudosenescence, observed is necessary to maintain tissue homeostasis following the injury.     

Rapid increase in plasma growth hormone after low-intensity resistance exercise with vascular occlusion.

Hormonal and inflammatory responses to low-intensity resistance exercise with vascular occlusion were studied. Subjects (n = 6) performed bilateral leg extension exercise in the seated position, with the proximal end of their thigh compressed at 214 +/- 7.7 (SE) mmHg throughout the session of exercise by means of a pressure tourniquet. Mean intensity and quantity of the exercise were 20% of 1 repetition maximum and 14 repetitions x 5 sets, respectively. In each set, the subjects repeated the movement until exhaustion. Plasma concentrations of growth hormone (GH), norepinephrine (NE), lacate (La), lipid peroxide (LP), interleukin-6 (IL-6), and activity of creatine phosphokinase (CPK) were measured before and after the exercise was finished and the tourniquet was released. Concentrations of GH, NE, and La consistently showed marked, transient increases after the exercise with occlusion, whereas they did not change a great deal after the exercise without occlusion (control) done at the same intensity and quantity. Notably, concentration of GH reached a level approximately 290 times as high as that of the resting level 15 min after the exercise. IL-6 concentration showed a much more gradual increase and was maintained at a slightly higher level than in the control even 24 h after exercise. Concentrations of LP and CPK showed no significant change. The results suggest that extremely light resistance exercise combined with occlusion greatly stimulates the secretion of GH through regional accumulation of metabolites without considerable tissue damage.     

Rapid increase of mitochondrial uncoupling protein and its mRNA in stimulated brown adipose tissue. Use of a cDNA probe

The administration of thyroxine to neonatal rats stimulates RNA synthesis by neuronal nuclei isolated from the developing rat brain cortex. Glial nuclei are relatively resistant to thyroxine treatment. The activity of neuronal RNA polymerase II is particularly stimulated by the hormone. Thyroxine also affects neuronal chromatin structure as shown by changes in the relative proportion of different subnuclear fractions obtained by gentle micrococcal nuclease digestion of nuclei from hormone-treated rats.     

Rapid increase in deep supercooling of xylem parenchyma

Malus pumila Mill. twigs were collected from September through December and stored at 5°C until the low temperature exotherms of the xylem were determined by differential thermal analysis. During the differential thermal analysis, cooling was interrupted, and temperatures of 5 to −18°C were held for 0.4 to 10 hours before cooling to −50°C was resumed. Control twigs were cooled to −50°C without interruption. Holding the twigs at 1.3 to −5°C shifted the start of the low temperature exotherm from about −20 to −30°C. Slightly higher (2.6°C) and lower (−10°C) temperatures were occasionally effective. The shift began within 20 to 30 minutes and increased progressively to 150 minutes. The acclimation was reversibly inhibited by N₂ atmosphere.     

Rapid increase of profilin-like proteins during liver development

Extraction of the cytoplasm of rat liver with 5% PCA and subsequent TCA (20%) treatment resulted in the recovery of only a few proteins, the predominant one having an apparent molecular weight of 12K. This 12K protein increased sharply (9-fold) between 2 weeks and 10-20 weeks and then decreased until at least 2 years after birth. 12K protein was separated into two components, 12K-1 (larger) and 12K-2 (smaller), by gel chromatography which had similar amino acid compositions to those of profilin (from Acanthamoeba and hog thyroid). Like profilin, 12K-1 protein inhibited the rate of actin polymerization, while 12K-2 protein did not. These findings suggest that liver profilin-like proteins increase sharply during development.     

Less is more! Rapid increase in plant species richness after reduced mowing in urban grasslands

Urban lawns provide space for recreation in cities, and they are an important part of urban green infrastructures. However, most lawns are intensively managed. As only few plant species can survive frequent mowing, urban lawns typically harbor only a limited number of plant species. To improve the biodiversity of urban lawns, it is often suggested to reduce the mowing frequency. Here, we studied the plant diversity of urban grasslands that have recently undergone management changes from mowing every few weeks to mowing only once or twice per season and compared them to intensively managed lawns. Within six years after the management changes, the grasslands with reduced mowing frequency hosted 30% more plant species than intensively managed lawns, and they were more heterogeneous both within and between grasslands. Additionally, the species composition of less frequently mown grasslands shifted from common mowing-tolerant lawn species to typical meadow species. Our study thus shows that the reduction of mowing is a simple and effective tool for increasing the biodiversity in urban grasslands.     

Kinetic parameters of calcium binding to tissue structures in human acute hypercalcemia

The kinetics of calcium binding by tissue structures was analyzed in 19 healthy volunteers (13 men and 6 women) from the age group of 33 ± 6.5 years under the conditions of acute hypercalcemia followed by drip intravenous infusion of calcium gluconate for 2.5 h. At the end of each 30-min period, the calcium amount retained by the tissue structures was recorded and the kinetic parameters of calcium binding were determined according to Langmuir and Scatchard. In all volunteers, there was a region of binding isotherm with positive cooperativity (linear regression in Scatchard’s coordinates) with the same buffer capacity (β_tis) for calcium in Langmuir’s coordinates (0.58 ± 0.24 L kg). Half of the volunteers exhibited cooperativity at [Ca²⁺] = 1.3–1.5 mmol/L, while others, at [Ca²⁺] = 1.0–1.3 mmol/L, which corresponded to the differences in the association constant (K_a) and the number of interactive sites (n) with [Ca²⁺] = 1 mmol/L. Additionally, two segments of the binding isotherm were detected with the successive binding of calcium to one set of non-interactive centers with similar kinetic parameters of calcium binding (β_tis, K_a, n). Four different types of calcium binding in healthy volunteers were found. The results of this study may serve as the basis for a functional diagnostic test of disorders of the tissue calcium-binding properties in different pathological conditions.     

Immunotoxic catecholamine lesions attenuate 2DG-induced increase of AGRP mRNA

Injections of the immunotoxin, saporin conjugated to anti-dopamine-beta-hydroxylase (DSAP), into the hypothalamic paraventricular nucleus (PVH) selectively destroy norepinephrine (NE) and epinephrine (E) terminals in the medial hypothalamus and abolish glucoprivic feeding. We utilized PVH DSAP injections to examine the role of NE/E neurons in the previously reported 2-deoxy-D-glucose (2DG)-induced increases in mRNA levels for the orexigenic peptides, AGRP and NPY. Northern blot analysis revealed that DSAP lesions elevated basal but blocked 2DG-induced increases in AGRP mRNA levels. Changes in NPY mRNA were not detectable. AGRP neurons may contribute to circuitry activated by NE/E neurons for elicitation of glucoregulatory responses.     

Rapid increase of vacuolar volume in response to salt stress

Suspension-cultured cells of mangrove [Bruguiera sexangula (Lour.) Poir.] showed a rapid increase in vacuolar volume under salt stress, although there was no change in the cell volume. The rapid increase in the vacuolar volume was an active process, which followed the activation of the tonoplast H(+)-ATPase and the vacuolar acid phosphatase. The same phenomenon was observed in barley (Hordeum vulgare L. cv. Doriru) root meristematic cells under salt stress but not in pea ( Pisum sativum L.). Increases in vacuolar volume could potentially protect the cytoplasm by decreasing the cytoplasmic volume during the initial phases of salt stress.     

Different patterns in the induction of metallothionein mRNA synthesis among isoforms after acute ethanol administration

The induction of metallothionein (MT) isoform synthesis was investigated in mouse cerebral cortex 18 h after oral ethanol administration. The expression of MT-I isoform mRNA increased in a dose-dependent manner after ethanol loading at doses between 2 g/kg (ethanol/body weight) and 8 g/kg. Lipid peroxide formation, measured as the amount of malondialdehyde-reactive substances, remained at the control level after all of the administered ethanol doses. The expression of MT-III isoform mRNA remained at the control level up until an ethanol loading dose of 4 g/kg and then finally increased to a significant level at a dose of 8 g/kg, which is almost the LD₅₀ for oral ethanol in mice. The different patterns of MT synthesis induction among MT isoforms suggests that the MT-I isoform, which is ubiquitous in mammalian tissues, plays a significant role as an antioxidant. On the other hand, the MT-III isoform, which has a limited tissue distribution, especially in the central nervous system, seems to be implicated in tissue repair and/or protection against critical tissue injury.     

Subtype-specific increase in G-protein alpha-subunit mRNA by interleukin 1 beta

The guanine nucleotide regulatory proteins (G-proteins) which are substrates for ADP-ribosylation by pertussis toxin (alpha i-1, alpha i-2, alpha i-3 and alpha o) transduce a variety of hormonal signals. Endothelial cells express mRNA for three alpha i subtypes although the level of alpha i-1 mRNA is very low. Interleukin 1 beta (IL 1 beta), a pleiotropic inflammatory mediator which stimulates a complex series of responses in human endothelial cells leading to increased coagulation and platelet adhesion, increases expression of one subtype of alpha i (alpha i-2) mRNA in human endothelial cells as determined by Northern blot analysis without affecting the level of mRNA for other alpha-subunits. These studies show that mRNA levels for alpha i subtypes are independently regulated, suggesting that there may be subtype specificity in the cell's requirements for the Gi class of signal-transducing proteins.