Miniaturization of whole live cell-based GPCR assays using microdispensing and detection systems

Cell-based beta-lactamase reporter gene assays designed to measure the functional responses of G-protein-coupled receptors (GPCRs) were miniaturized to less than 2 microL total assay volume in a 3456-well microplate. Studies were done to evaluate both receptor agonists and antagonists. The pharmacology of agonists and antagonists for target GPCRs originally developed in a 96-well format was recapitulated in a 3456-well microplate format without compromising data quality or EC(50)/IC(50) precision. These assays were employed in high-throughput screening campaigns, allowing the testing of more than 150,000 compounds in 8 h. The instrumentation used and practical aspects of the assay development are discussed.     

Morphology and ultrastructure of reproductive organs of <Emphasis Type="Italic">Monocercus arionis</Emphasis> (Sibold, 1850) Villot, 1982 (Cestoda: Cyclophyllidea)

The morphology of the reproductive system at different stages of ontogenesis and ultrastructural peculiarities of the copulatory organs and uterine in Monocercus arionis (Sibold, 1850) Villot, 1982 (Cyclophyllidea) have been studied. The muscle of the outer wall of the cirrus bag has a higher organizational level than typical smooth-muscle cells in cestodes. The cirrus is armed with typical filamentous and bladelike microtriches. The uterine epithelium contacts the thin capsule of the developing embryos located closely to the uterus wall, and the embryos contact each other in the uterine cavity in what can be interpreted as placental interactions. The specificity of the structure and arming of the copulatory apparatus has been considered, and the ultrastructural peculiarities of the uterus in the members of different orders of cestodes have been compared and analyzed.     

Development of the energetics of muscular exercise with age: Summary of a 30-year study: II. “Power zones” and their changes with age

An ergometric model of relative power zones of cyclic muscular exercise is presented. Age-related changes in the main ergometric parameters, as well as the structure of the entire range of muscle loads bearable by humans, have been studied in 7-to 17-year-old boys. The ergometric parameters reflecting the power characteristics of muscles increase by a factor of about two in the age interval studied, while the parameters characterizing the capacity of the energy system of skeletal muscle increase by a factor of several tens. This reflects the actual increase in muscle working capacity during the maturation of the musculoskeletal system.     

Intraretinal variability and specialization of cones in Japanese anchovy (Engraulis japonicus,Engraulidae)

The retina of anchovies is characterized by an unusual arrangement and ultrastructure of cones. In the retina of Japanese anchovies, Engraulis japonicus, three types of cones are distributed into rows. The nasal, central, temporal, and ventro‐temporal regions of the retina were occupied exclusively by the long and short cones. Triple cones, made up of two lateral components and one smaller central component, were found only in the dorsal and ventro‐nasal retinal regions. In the outer segments of all short and long cones from the ventro‐temporal region, the lamellae were oriented along the cell axis and were perpendicular to the lamellae in the long cones, providing a morphological basis for the detection of polarization. This lamellar orientation is unique to all vertebrates. The cones were examined with respect to regional differentiation in their size and spectral properties via light microscopy, transmission electron microscopy, and microspectrophotometry. Various dimensions of cones were measured in preparations of isolated cells. The cones from the ventro‐temporal region had different dimensions than cones of the same type located in other retinal regions. Triple cones from the dorsal region were significantly larger than triple cones from the ventro‐nasal region. The spectral absorbance of the lateral components of triple cones in the ventro‐nasal retina was identical to the absorbance of all long and short cones from the ventro‐temporal region. These are shifted to shorter wavelengths relative to the absorbance of the lateral components of the triple cones located in the dorsal retina. Thus, the retina of the Japanese anchovy shows some features of regional specialization common in other fishes that improves spatial resolution for the upwards and forwards visual axis and provides spectral tuning in downwelling light environment. That results from the differentiation of cone types by size and by different spectral sensitivity of various retinal areas. J. Morphol. 277:472–481, 2016. © 2016 Wiley Periodicals, Inc.     

Development of the energetics of muscular exercise with age: Summary of a 30-year study. III. Endogenous and exogenous factors influencing the development of the energetics of skeletal muscles

Differences in the age-related changes in the physical working capacity and the structure of the energy supply of muscular exercise of different powers determined by external (physical education conditions) and internal (gender characteristics) factors are described. Periods of synchronous and asynchronous development of the physical working capacity parameters have been revealed that are characterized by predomination of endogenous or exogenous influences on the processes studied.     

Seasonal density dynamics and distribution of brachyuran crab larvae (Decapoda: Brachyura) in Amursky and Ussuriysky bays, Sea of Japan

The species composition, period of occurrence, density, and distribution of brachyuran crab larvae (Decapoda: Brachyura) in the Amursky and Ussuriysky bays (Sea of Japan) were studied in May–October 2007 and 2008. The larvae of this group were not numerous, with peak densities occurring in July at maximum water temperatures. The average brachyuran larvae density was up to 126.9 ind./m³ in Ussuriysky Bay and 68 ind./m³ in Amursky Bay; the contribution of Brachyura to all decapod larvae reached 80–90%. In Amursky Bay, the highest density of brachyuran larvae (up to 584 ind./m³) was observed along the western coast over depths to 20 m; in Ussuriysky Bay, the larval density was highest (up to 1817 ind./m³) in the shallowwater northern part and progressively decreased with depth. Larvae of 18 brachyuran species were found in plankton samples. In Amursky Bay, intertidal crab larvae of the genus Hemigrapsus dominated, Eriocheir japonica larvae were fairly numerous. Larvae of the pea crabs Pinnixa rathbuni and Tritodynamia rathbunae and zoeae of the bigtooth rock crab Glebocarcinus amphioetus were predominant in Ussuriysky Bay.     

Matricide in Caenorhabditis elegans as an example of programmed death of whole animal organism: role of mitochondrial oxidative stress

Nematodes Caenorhabditis elegans is a widely used model for studying the genetic and molecular mechanisms that determine the lifespan. The choice between the two vital program strategies of adult hermaphrodite C. elegans--normal aging and matritcide (programmed death), is largely affected by the availability of food, and also depends on a variety of stresses. We decided to test the hypothesis that, in line with the phenoptosis theory, oxidative stress increases probability of the programmed death of the whole organism. It is shown that high concentrations of paraquat (strong mitochondrial stress) significantly increase the propensity to matricide. In this case, mutants with a reduced antioxidant capacity of mitochondria (nnt) are more sensitive to the reagent. On the other hand, the concentrations of paraquat, necessary for the manifestation of this effect, are toxic to the offspring, while at low concentrations matricide of mutant worms and wild-type worms occurs with equal frequency. Therefore it is safe to conclude that oxidative stress is not the key initiating mechanism of matricide under normal conditions.     

Structural and functional features of gill epithelium and the brood pouch of pipefish <Emphasis Type="Italic">Sygnathus acusimilis</Emphasis> (Syngnathidae,Gasterosteiformes) during adaptation to dilute sea water

Study of the dilate sea-water tolerance of the pipefish Syngnathus acusimilis indicated that it successfully acclimatized to an abrupt change in water salinity from 5 to 32‰. Larvae in the brood pouch also successfully acclimatized to salinity change, and, after several days, juveniles left the brood pouch and continued to live. In freshwater, adult and fry perished during several minutes. In the gill epithelium of S. acusimilis, chloride cells of only a marine type were found. A comparison of the ultrastructure of chloride cells of gill epithelium and mitochondrion-rich cells of the brood pouch of S. acusimilis demonstrated their high similarity. The ultrastructure of cells of the brood pouch indisputably attests to their involvement in maintaining an osmotic and ionic homeostasis in the cavity of the brood pouch.