Adrenergic receptor characteristics of cardiac myocytes cultured in serum-free medium: comparison with serum-supplemented medium

Neonatal rat ventricular myocytes cultured in serum-free medium coexpress both alpha 1 and beta 1 receptors as determined by radioligand binding studies. In cells exposed to serum for 48 hr surface area increased 3.69 fold, but the maximum number of binding sites ([125I]-iodocynanopindolol) only increased 1.5 fold from 12956 +/- 7579 to 19676 +/- 5181 sites/cell (n = 5, p less than .05) yielding a value of 2.48 sites/um2 for cells grown in serum-supplemented medium compared with 6.96 sites/um2 for cells grown in serum-free medium. Thus serum-induced hypertrophy is associated with a decrease in beta 1 receptor density relative to cell size; however, adenylate cyclase response is unaffected. This cell culture system constitutes an excellent model for studying interventions that may influence the regulation of cardiac myocyte hypertrophy by nonhemodynamic factors, particularly through the adrenergic receptor system.     

Gestational exposure to ethane dimethanesulfonate permanently alters reproductive competence in the CD-1 mouse

Although the adult mouse Leydig cell (LC) has been considered refractory to cytotoxic destruction by ethane dimethanesulfonate (EDS), the potential consequences of exposure during reproductive development in this species are unknown. Herein pregnant CD-1 mice were treated with 160 mg/kg on Gestation Days 11-17, and reproductive development in male offspring was evaluated. Prenatal administration of EDS compromised fetal testosterone (T) levels, compared with controls. EDS-exposed pups recovered their steroidogenic capacities after birth because T production by hCG-stimulated testis parenchyma from prepubertal male offspring was unchanged. However, prepubertal testes from prenatally exposed males contained seminiferous tubules (STs) devoid of germ cells, indicating a delay in spermatogenesis. In adults, some STs in exposed males still contained incomplete germ cell associations corroborating observed reductions in epididymal sperm reserves, fertility ratios, and litter size. Morphometry revealed an EDS-induced increase in interstitial area and a concomitant decrease in ST area, but stereology revealed an unexpected decrease in the number and size of the LCs per testis in exposed males. Paradoxically, there was an increase in both serum LH and T production by adult testis parenchyma, indicating that the LCs were hyperstimulated. These data demonstrate permanent lesions in LC development and spermatogenesis caused by prenatal exposure in mice. Thus, although adult mouse LCs are insensitive to EDS, EDS appears to have direct action on fetal LCs, resulting in abnormal testis development.     

Effects of aging on the cardiac remodeling induced by chronic high-altitude hypoxia in rat

Effects of chronic high-altitude hypoxia on the remodeling of right ventricle were examined in three age groups of rats: 2, 6, and 18 mo. The extent of right ventricular (RV) hypertrophy (RVH) showed an age-associated diminution. RV cell size and pericellular fibrosis showed a significant increase in the 2- and 6-mo-old exposed rats but not in the 18-mo-old exposed rats compared with control. A hyperplasic response was underscored in the three exposed age groups but appeared less pronounced in the 18-mo-old rats. A significant decrease in the transient outward potassium current (Ito) density was observed in RV cell only in the 2-mo-old exposed group compared with the control group. In the control group, there was a clear tendency for Ito density to decrease as a function of age. The sustained outward current density was modified neither by the hypoxia condition nor by the age. Neither the cytochrome c oxidase activity nor the heat shock protein 72 content in the RV was altered after hypoxic exposure regardless of age. The norepinephrine content in the RV was significantly decreased in each age group exposed to hypoxia when compared with their age-matched control group. Our findings indicate that the remodeling (at morphological and electrophysiological levels) induced by chronic hypoxia in the RV can be decreased by the natural aging process.     

Role of intracellular calcium and sodium in light adaptation in the retina of the honey bee drone (Apis mellifera, L)

In the honey bee drone, the decrease in sensitivity to light of a retinula cell exposed to background illumination was found to be accurately reflected by the difference in amplitude between the initial transient depolarization and the lowest steady depolarization evoked by the background light. It is shown that both the decrease in sensitivity to light and the accompanying drop in potential from the transient to the plateau can be prevented by injecting EGTA intracellularly. A decrease in duration and amplitude of responses to short test flashes such as observed immediately after illumination was found to occur too when Ca or Na, but not K, Li, or Mg injected into dark-adapted retinula cells. Injection of EGTA into a retinula cell maintained a steady state of light adaptation, was found to cause an increase in amplitude and duration of the response to a short test flash, thus producing the effects of dark adaptation. It is suggested that, in the retina of the honey bee drone, an increase in intracellular calcium concentration plays a central role in light adaptation and that an increase in intracellular sodium concentration, resulting from the influx of sodium ions during the responses to light, could lead to this increase in intracellular free calcium.     

Vertical distribution of coralline algae in the rocky intertidal of northern Chile

Studies on crustose corallines present in the intertidal region at three localities in northern Chile (30°S), show that these algae are well represented in exposed and protected sites, reaching up to 90% cover. Species composition differs between sites with the common occurrence of species or morphological variants of Spongites and a single taxon attributed to Phymatolithon at the most exposed sites, and species of Lithophyllum and Titanoderma at more protected localities. The two Lithophyllum taxa recorded are distinguished by the presence/absence of protuberances, cell size and degree of calcification, while Titanoderma taxa are segregated by the thickness of the thallus, hypothallic and perithallic cell size and shape. Spongites taxa are distinguished on the basis of external morphology and anatomical features such as cell size, degree of calcification and percentage of fusions between cells. The variability of these features within each species is still unknown thus, the taxa remain without specific epithet until further studies. Examination of specific types recorded for nearby regions are also required in order to clarify the taxonomy of the group in these coasts.     

结核杆菌的药物外排泵

外排泵基因过度表达促使细胞内的药物转移到细胞外,胞内药物浓度下降,致使药物临床疗效降低或无效。外排泵上有多个与识别底物结构无关的阴离子凹穴(底物结合位点),通过静电引力非特异性结合各种含阳离子的药物,在能量协助下将药物排到细胞外,进而导致细菌抗药性的产生。研究表明,结核杆菌基因组中存在外排泵基因,但仅分离到几个有功效的外排泵。本文就相关研究进展进行综述。     

A unique amino acid substitution in the outer membrane protein OmpA causes conjugation deficiency in Escherichia coli K-12

The outer membrane protein OmpA of E. coli K-12 can serve as a receptor for phages and is required for stabilizing mating aggregates during F'-mediated conjugation. Selection for resistance to OmpA-specific phages yields mutants with alterations in the protein at four cell surface exposed sites. It is shown that conjugation deficiency can be caused by apparently only one type of amino acid substitution at one of these sites, the replacement of glycine-154 by aspartic acid. This suggests that, in contrast to binding of phages, a ligand of the donor cell recognizes only a very small area of the protein.     

Metabolism of isolated fat cells from various tissue sites in the rat: influence of hemorrhagic hypotension

The in vitro lipolytic response to norepinephrine by rat adipocytes from epididymal, subcutaneous, perirenal, mesenteric, and omental tissue sites was studied in control and hypotensive animals. Lipolysis per millimole of triglyceride was found to be three to four times higher in mesenteric and omental fat cells than in adipocytes of the other sites sampled. The high lipolytic activity of mesenteric and omental adipocytes was partly attributable to their smaller cell size; however, lipolysis per cell was also higher. Hemorrhagic hypotension caused a 50-60% decrease in lipolytic activity at four of the five sites studied. Adipocytes of omental origin maintained their lipolytic activity at the prehypotensive level, however, indicating that the metabolic adjustments brought about by hemorrhagic hypotension are not uniform at all adipose tissue sites.     

Alterations in surface ultrastructure and anionic sites of rat dimethylhydrazine-induced intestinal tumors

The relative roles of cell surface shedding and electronegative charge as determinants of metastatic capacity were studied in experimentally produced intestinal tumors. The ultrastructural organization and distribution of anionic sites on the luminal plasma membrane surface components were examined in small intestinal and colonic tumors induced in male Sprague-Dawley rats with 1,2-dimethylhydrazine. The overall distribution of negatively charged groups was demonstrated with ruthenium red staining. Compared to normal epithelial cells, neoplastic cells revealed evidence of decreased cell surface shedding as manifested by decreased numbers of membrane-bound bodies, and an increased quantity of glycocalyx. Malignant cell surfaces were directly exposed to the intestinal lumen as a result of losing the enteric surface coat covering. The exposed microvilli appeared damaged with shortening and blunting. The glycocalyx and surface coat both reacted strongly with ruthenium red indicating the presence of anionic sites. As a result of surface coat loss, the malignant cell surface components revealed an overall decrease in net negative charge. These alterations in cell surface component ultrastructure and electronegative charge appear to be consistent with the low capacity for chemically induced rat intestinal tumors to metastasize.     

Ferrocyanide enhancement of concanavalin A-ferritin and cationized ferritin staining blood cell surface glycoconjugates

Synopsis Ferrocyanide was used to enhance cationized ferritin and concanavalin A-ferritin (Con A-ferritin) staining of surface glycoconjugates of peripheral blood and bone marrow cells from rabbits and humans. The glutaraldehyde-fixed cells were stained with Con A-ferritin or cationized ferritin and then exposed to a ferrocyanide solution. The resulting cuboidal and irregular stain deposits averaged 50 nm in diameter when viewed with the transmission (TEM) and scanning electron microscope (SEM). Rabbit blood cells demonstrated more Con A binding sites than human blood cells and the decrease in binding sites observed with maturation of human granulocytic and erythrocytic cells was not evident in rabbit cells. Differences in binding of cationized ferritin to rabbit and human cell surfaces were less prominent than that observed for Con A. These results extend previous studies of blood cell surface glycoconjugates and demonstrate that ferrocyanide enhancement significantly facilitates SEM evaluation of Con A-ferritin and cationized ferritin bound to cell surfaces.     

Ultrastructural changes induced by low concentrations of DDT in the livers of the zebrafish and the guppy

Following exposure to 0.3–3.0 ppb DDT, the livers of zebrafish and guppies were examined by electron microscopy. After 24 h in 1.0 ppb or 72 h in 0.3 ppb, zebrafish had a complete loss of glycogen, decrease in cell size and a cytoplasmic compartment filled with rough endoplasmic reticulum (ER). Guppies exposed for periods of up to 28 days showed only a gradual accumulation of smooth ER and no change in glycogen, lipid, rough ER or cell size. The relationship of these changes to the zebrafish's sensitivity and the guppy's tolerance to DDT is dicussed.     

Proliferation-associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture.

The oxygen consumption rate, proliferative activity, and morphology of EMT6/Ro mouse mammary sarcoma cells in monolayer and multicellular spheroid culture have been investigated in a comparative study. During the transition of monolayer cells from the exponential into the plateau growth phase, there is a distinct decrease in the cellular volume that is associated with a corresponding decrease in the proliferative and respiratory activity of the cells. The decline in cell volume is mainly due to a decrease in the content of cytoplasm, whereas the size of the nucleus is only slightly reduced. A concomitant decrease in the number of mitochondria per cell obviously accounts for the reduction in cellular oxygen uptake. Despite a continuous decrease of cell proliferation from the surface to interior regions of EMT6 spheroids reflected by a gradient in tritiated thymidine labeling, volume-related oxygen consumption is rather uniform in viable regions of these aggregates. The finding can be explained by the results of the morphometric evaluation showing a uniform volume density of mitochondria, i.e., of oxygen-consuming sites within these spheroids.     

Effects of the cationic protein poly-L-arginine on airway epithelial cells in vitro

BACKGROUND: Allergic asthma is associated with an increased number of eosinophils in the airway wall. Eosinophils secrete cationic proteins, particularly major basic protein (MBP). AIM: To investigate the effect of synthetic cationic polypeptides such as poly-L-arginine, which can mimic the effect of MBP, on airway epithelial cells. METHODS: Cultured airway epithelial cells were exposed to poly-L-arginine, and effects were determined by light and electron microscopy. RESULTS: Poly-L-arginine induced apoptosis and necrosis. Transmission electron microscopy showed mitochondrial damage and changes in the nucleus. The tight junctions were damaged, as evidenced by penetration of lanthanum. Scanning electron microscopy showed a damaged cell membrane with many pores. Microanalysis showed a significant decrease in the cellular content of magnesium, phosphorus, sodium, potassium and chlorine, and an increase in calcium. Plakoglobin immunoreactivity in the cell membrane was decreased, indicating a decrease in the number of desmosomes CONCLUSIONS: The results point to poly-L-arginine induced membrane damage, resulting in increased permeability, loss of cell-cell contacts and generalized cell damage.     

Storage and decomposition of particulate organic matter along the longitudinal gradient of an agriculturally-impacted stream

An agriculturally-impacted stream in northern Idaho was examined over a two-year period to determine seasonal and longitudinal patterns of the storage and decomposition of particulate organic matter. Biomass of benthic organic matter (BOM) was considerably less than values reported in the literature for comparable, undisturbed streams. Coarse, fine, and total benthic particulate organic matter were not correlated with parameters pertaining to stream size (e.g., stream order), but were correlated with sample site and amount of litterfall. The association of BOM with site and litterfall suggests that storage of particulate organic matter is a function of local characteristics rather than stream size. Low biomass of stored organic matter is a response to the low input of terrestrially-derived organic matter resulting from removal of climax vegetation.Leaf packs of alder, Alnus sp., were placed in the stream seasonally for 30 and 60 d. While there were significant differences for months, there was no significant difference among sites for leaf packs exposed for 30 d. Significant differences were observed among both sites and months for leaf packs exposed for 60 d; however, differences among sites accounted for only 5% of the variance. The absence of differences in decomposition of organic matter along the gradient of Lapwai Creek, despite heterogeneity of the drainage basin and availability of organic matter, may be in response to the overall low biomass of stored benthic organic matter. This study demonstrates that agricultural activity can substantially influence instream heterotrophic processes through reduced availability of organic matter and can shape community structure and ecosystem dynamics of streams flowing through agricultural drainage basins.     

Stabilization of the adenylate energy charge in erythrocytes of rats and humans at high altitude hypoxia.

1. Stabilization of adenylate energy charge and control of adenylate pool were analysed in the erythrocytes of the rat and the human exposed to highly hypoxic conditions. 2. Red cell energy charge was decreased in the rats exposed to a simulated altitude of 5000-8000 m, and then recovered to the normal value with the depletion of adenylate pool. 3. The energy charge and the adenylate pool size of the human erythrocytes did not show any change under highly hypoxic conditions. 4. Anaerobic incubation of rat erythrocytes caused a marked decrease in the energy charge, and its recovery was accompanied by the depletion of total adenylates. 5. The energy charge and total adenylates of human red cells did not change under the anaerobic incubation of erythrocytes. 6. These results suggest that the energy charge of rat erythrocytes can be controlled by depletion of the adenylate pool, but the adenylate degradation is not responsible for the stabilization of the energy charge in human erythrocytes.     

Inhibition by dexamethasone of beta 3-adrenergic receptor responsiveness in 3T3-F442A adipocytes. Evidence for a transcriptional mechanism.

Modulation of beta 3-adrenergic receptor (beta 3AR) expression by dexamethasone was investigated in the murine 3T3-F442A adipocytic cell line. In untreated cells, a major population of binding sites (62,000-114,000 sites/cell) of low affinity for (-)-[3H] CGP12177 and (-)-[125I]iodocyanopindolol (corresponding to the beta 3AR subtype) was present along with a minor population (6,500-8,000 sites/cell) of sites of high affinity for the radioligands (corresponding to a mixture of the beta 1 and beta 2AR subtypes). Long-term exposure of the cells to 250 nM dexamethasone led to a sharp decrease in beta 3AR density (less than 5,000 sites/cell) which paralleled a diminished potency of the beta 3AR-selective agonists BRL37344 and CGP12177 to stimulate the production of intracellular cAMP. Analysis of RNA by polymerase chain reaction and nuclear run-on assays indicated that dexamethasone inhibited the synthesis of beta 3AR mRNA, resulting in 4-8-fold decrease in the steady-state levels of this mRNA. The down-regulation of beta 3AR protein and cellular mRNA appeared to be mediated by the receptor for glucocorticoids as assessed by the antagonistic action of the anti-glucocorticoid RU38486.     

Relationship between changes in buoyant density and formation of new sites of cell wall growth in cultures of streptococci (Enterococcus hirae ATCC 9790) undergoing a nutritional shift-up.

When the glutamate concentration of cultures of Enterococcus hirae was raised from 20 to 300 micrograms/ml, the mass doubling time decreased from ca. 85 to 45 min in 9 min, but balanced growth was not reestablished for 30 to 40 min. During the unbalanced period of growth, RNA and protein synthesis proceeded more rapidly than did peptidoglycan synthesis, buoyant density increased from ca. 1.1024 to 1.1075 g/ml, and the rate of formation of new cell wall growth sites transitorily accelerated above the new growth rate. When studied as a function of cell size, all cultures showed buoyant density to decrease around cell separation, increase as cells increased in size, and then plateau when cells reached large volumes. Greater increases in buoyant density as a function of cell size were seen after shift-up, with the greatest increases observed at 15 to 20 min after shift-up, when the rate of formation of new sites was also maximal. In a population of cells examined by electron microscopy 15 min after shift-up, buoyant density and the frequency of cells with new sites increased as old sites approached the size of two poles. These data were consistent with a model whereby buoyant density increases in the terminal stages of the cell cycle when the surface grows slower than the cytoplasm. The greater the difference in the rates of inside to outside growth, the greater the increase in buoyant density and the more frequently new sites will be initiated.     

Quantal analysis of synaptic processes in the neocortex

The application of fluctuation analysis to studies of synaptic function in the neocortex is discussed. Analysis of failures of transmission has been valuable in indicating whether a presynaptic or a postsynaptic site is responsible for a change in synaptic efficacy. When combined with detailed ultrastructural verification of all synapses involved in an individual cell to cell connection, a reasonable estimate of quantal size and release probability under conditions of low frequency activity can be obtained. However, both the number of available release sites in functional terms and the probability that an action potential (AP) will release transmitter from any given site can vary from AP to AP at higher frequencies. A variety of presynaptic mechanisms that modulate release are now apparent. For example, one mechanism dominates release patterns at one class of connection which is insensitive to absolute firing frequency, but responsive to changes in frequency. At another class of connection, a different mechanism dominates, resulting in high sensitivity to frequency.     

Role of mitochondria in reactive oxygen species generation and removal; relevance to signaling and programmed cell death

Reactive oxygen species (ROS) are universal products of aerobic metabolism, which can be also produced in stress conditions. In eukaryotic cells, mitochondria are the main source of ROS. The main mitochondrial sites of ROS formation are electron carriers of respiratory chain. However, there are also other enzymatic sites capable of ROS generation in different mitochondrial compartments. Reactive oxygen species can cause serious damage to many biological macromolecules, such as proteins, lipids and nucleic acids, which oxidation leads to a lost of their biological properties and eventually to a cell death. Mitochondria, which are also exposed to harmful ROS action, have a defense system that decreases ROS production (first line of defense) or removes generated ROS (second line of defense). Mitochondrial antioxidant system involves proteins that decrease ROS formation, enzymes that directly react with ROS, and non-enzymatic antioxidants that also remove ROS and other oxygen derivatives. Mitochondrial ROS can also act as signal messengers and modify operation of many routes in different cell compartments. Mitochondrial ROS are also important in execution of programmed cell death.     

Does a warmer lake mean smaller benthic algae? Evidence against the importance of temperature–size relationships in natural systems

Ectotherms decrease in size with increasing ambient temperature. Temperature–size relationships (TSR) have been observed experimentally in a wide range of animals, algae, protozoans and bacteria. However, it is still unclear whether temperature is an important factor controlling the size of organisms in natural populations. In this study, we used natural variability in water temperature in the nearshore areas of a single lake to test TSR in populations of benthic diatoms. We deployed standard tile substrates at 5 m depth (similar light availability) at cold and warm sites that were exposed to different hydrodynamic forces. We compared cell sizes of three species of diatoms (Achnanthidium minutissimum, Gomphonema acuminatum and Gyrosigma acuminatum) at these sites. Counter to the TSR, diatom cells at warm sites were either larger (Achnanthidium, Gomphonema) or similar in size (Gyrosigma) compared to those at colder sites. Diatom size was also related to site exposure (hydrodynamic forces), but differently for species with different architectures. TSR were not detectable in the field for these three species of benthic diatom, even when tested within a single ecosystem at a given time of the year. The size of benthic diatoms, however, varied in a predictable way between sites, and such differences could affect the functioning of these primary producers in different parts of the littoral zone.