Integral role of the I′-helix in the function of the “inactive” C-terminal domain of catalase–peroxidase (KatG)

Catalase–peroxidases (KatGs) have two peroxidase-like domains. The N-terminal domain contains the heme-dependent, bifunctional active site. Though the C-terminal domain lacks the ability to bind heme or directly catalyze any reaction, it has been proposed to serve as a platform to direct the folding of the N-terminal domain. Toward such a purpose, its I′-helix is highly conserved and appears at the interface between the two domains. Single and multiple substitution variants targeting highly conserved residues of the I′-helix were generated for intact KatG as well as the stand-alone C-terminal domain (KatG^C). Single variants of intact KatG produced only subtle variations in spectroscopic and catalytic properties of the enzyme. However, the double and quadruple variants showed substantial increases in hexa-coordinate low-spin heme and diminished enzyme activity, similar to that observed for the N-terminal domain on its own (KatG^N). The analogous variants of KatG^C showed a much more profound loss of function as evaluated by their ability to return KatG^N to its active conformation. All of the single variants showed a substantial decrease in the rate and extent of KatG^N reactivation, but with two substitutions, KatG^C completely lost its capacity for the reactivation of KatG^N. These results suggest that the I′-helix is central to direct structural adjustments in the adjacent N-terminal domain and supports the hypothesis that the C-terminal domain serves as a platform to direct N-terminal domain conformation and bifunctionality.     

Microsatellite assessment of the genetic diversity in indigenous populations of curimba (<Emphasis Type="Italic">Prochilodus argenteus</Emphasis>) in the São Francisco river (Brazil)

The freshwater fish curimba (Prochilodus argenteus) is an endemic species of the São Francisco River basin, with great aquaculture potential. The construction of several hydroelectric power dams along the river, and the pulsed releases of water from these dams altered and degraded the native fish habitats, which led to the development of a hatchery-reared fingerling restocking program. A comprehensive genetic baseline evaluation of the indigenous populations of curimba and of a hatchery stock was done using microsatellite markers. Pairwise F _ST values, Bayesian analysis and the number of migrants per generation were used to illustrate the genetic structure of the curimba populations along almost 3000 km of the river, suggesting the existence of three units of conservation: one in the Upper, a second in the Middle and a third including part of the Middle, Submiddle and Lower stretches. We have analyzed the genetic contribution of hatchery stock releases in two stretches of the river using individual admixture coefficients; and the results showed that admixture makes a negligible contribution to indigenous recruitment. We discussed about supplementation programs for this river from the perspective of three units of conservation and the risks associated with using domesticated fish.     

Induction of androgenetic development of the brook charr (Salvelinus fontinalis) × Arctic charr (Salvelinus alpinus) hybrids in eggs derived from the parental species

Failure of interspecific androgenesis between brook charr (Salvelinus fontinalis, Mitchill 1814) and Arctic charr (Salvelinus alpinus, L.) has been attributed to the conflict between the egg cytoplasm of one species and the sperm nucleus of the other species. To overcome this incompatibility, sperm derived from the brook charr × Arctic charr hybrid male was used to induce androgenetic development in eggs originating from the parental species as well as their hybrids. The eggs were subjected to 420 Gy of X-radiation to damage the maternal nuclear DNA and inseminated with untreated sperm. Haploid zygotes were exposed to high hydrostatic pressure shock (7000 psi for 4 min), which was applied 420 min after insemination to inhibit the first cell cleavage and recover the diploid state of the zygote. The androgenetic diploid offspring that hatched from the brook charr, the Arctic charr and the hybrids eggs had survival rates of 4.7 ± 0.6%, 1.2 ± 0.4% and 16.8 ± 0.5%, respectively. Drastic mortality among the hatched androgenetic individuals was observed within the first five months of rearing. Cytogenetic analysis of the androgenetic progenies exhibited residues of the irradiated maternal nuclear genome in the form of radiation-induced chromosome fragments in 47% of the specimens that were examined. Interactions between the egg cytoplasm and the sperm nucleus, the low quality of the gametes, the expression of homozygous paternal lethal alleles and the incomplete inactivation of the maternal chromosomes were identified as factors responsible for the large mortality among androgenetic embryos and hatchlings.     

Larvae of the invasive species <Emphasis Type="Italic">Limnoperna fortunei</Emphasis> (Bivalvia) in the diet of fish larvae in the Paraná River,Argentina

To assess the trophic impact of the planktonic larvae of the invasive Asian bivalve Limnoperna fortunei, introduced in South America around 1990, we investigated the gut contents of fish larvae collected at monthly intervals between October 2000 and March 2001 at three locations along a 600 km stretch of the Paraná River, and during November 2004 in two areas of marginal lagoons connected to the river near the city of San Nicolás, Argentina. Zooplankton was also collected in the lagoons in 2004. In total, 11,956 fish larvae were retrieved, of which 1,511 were used for detailed analyses. Of the 15 fish taxa collected, 11 had veligers of L. fortunei in their gut. Fish larvae with empty guts represented 60% (San Nicolás) to 72% (Paraná River) of the total number of fish. Proportions of feeding fish larvae with L. fortunei veligers in their guts varied between 20% (San Nicolás) and 56% (Paraná River); in 15% of the guts analyzed, L. fortunei was the only food item recorded. For those specimens that had consumed L. fortunei larvae and any other food item, L. fortunei was the most important item in 55% (Paraná River) to 71% (San Nicolás) of the animals in terms of biomass. No major temporal or spatial changes in the diet were observed along the Paraná River, but the relative biomass contribution of L. fortunei larvae differed strongly in fishes of different developmental stage. In protolarvae and mesolarvae, veligers accounted for 30–35% of the gut contents. In metalarvae, veligers accounted for only 3%, indicating enhanced food supply for the earliest fish life stages. Comparison of the relative proportions of the three main zooplankton types (L. fortunei veligers, cladocerans, and copepods) in the water and in larval fish guts indicates that L. fortunei is always selected positively over the other two prey types. While our results strongly suggest that the expansion of L. fortunei results in an enhanced food supply for local fish populations, they do not necessarily imply that the overall effect on the ecosystem in general, and on the fish fauna in particular is beneficial.     

Structure and Location of the Regulatory β Subunits in the (αβγδ)4 Phosphorylase Kinase Complex

Phosphorylase kinase (PhK) is a hexadecameric (αβγδ)₄ complex that regulates glycogenolysis in skeletal muscle. Activity of the catalytic γ subunit is regulated by allosteric activators targeting the regulatory α, β, and δ subunits. Three-dimensional EM reconstructions of PhK show it to be two large (αβγδ)₂ lobes joined with D₂ symmetry through interconnecting bridges. The subunit composition of these bridges was unknown, although indirect evidence suggested the β subunits may be involved in their formation. We have used biochemical, biophysical, and computational approaches to not only address the quaternary structure of the β subunits within the PhK complex, i.e. whether they compose the bridges, but also their secondary and tertiary structures. The secondary structure of β was determined to be predominantly helical by comparing the CD spectrum of an αγδ subcomplex with that of the native (αβγδ)₄ complex. An atomic model displaying tertiary structure for the entire β subunit was constructed using chemical cross-linking, MS, threading, and ab initio approaches. Nearly all this model is covered by two templates corresponding to glycosyl hydrolase 15 family members and the A subunit of protein phosphatase 2A. Regarding the quaternary structure of the β subunits, they were directly determined to compose the four interconnecting bridges in the (αβγδ)₄ kinase core, because a β₄ subcomplex was observed through both chemical cross-linking and top-down MS of PhK. The predicted model of the β subunit was docked within the bridges of a cryoelectron microscopic density envelope of PhK utilizing known surface features of the subunit.     

Parastenocaris andreji n. sp. (Crustacea; Copepoda) – the first record of the genus in Slovenia (SE Europe)

Parastenocaris andreji n. sp. was found in ground water in a northwestern part of Slovenia (southern part of Central Europe). Two adult females of the new species were filtered out of 20 000 l of water during examination of an aquifer for water supply. The bore well was 10 m deep. The aquifer is fed by rainwater and probably only occasionally from a nearby alpine river Sava. The new species differs markedly from other known species of the genus by shape and armature of furcal rami, very long and slim maxillae and some characters on endopodites P2–P5. The closest related species are probably Parastenocaris nolliKiefer, 1938 with subspecies P. nolli alpinaKiefer, 1960, P. gertrudae Kiefer, 1968 and P. austriacaKiefer, 1976.     

Study of interactions between wild soybean subpopulations (<Emphasis Type="Italic">Glycine soja</Emphasis>) in the valley of the Tsukanovka river in the south of Russia’s Far East

Comparative analysis of the genetic structure of natural and anthropogenic populations of G. soja gives significant information about the formation of different populations and allows for the developing of measures for the preservation of the unique natural gene bank of wild soybean, which is a species closely related to cultivated soybean. In this study, ISSR-markers were used to carry out a comparative analysis of the genetic structure of natural and anthropogenic subpopulations of G. soja for studying the possible mutual influence of subpopulations of anthropogenic and natural phytocenosis on the formation of their genetic diversity and studying the genetic structure of natural subpopulations of wild soybean in the contact places between the two types of cenoses. As a result, the characteristics that describe the genetic diversity of the studied populations have been identified, and the important role of interaction between subpopulations of different phytocenoses in the formation of the spatial genetic structure of the population in the Tsukanovka river valley have been demonstrated.     

Characteristics of photosynthesis in maize leaves (С<Subscript>4</Subscript> plants) upon changes in the level of illuminance and nitrate nutrition

We studied assimilation of ¹⁴СО₂ and distribution of ¹⁴С among the products of 3-min-long photosynthesis of maize (Zea mays L.) leaves. The day before the experiment, half of the plants were fertilized with Ca(NO₃)₂ (1 g/L of water) at a rate of 6 L/m². Five days before the experiment, some plants were shaded for adaptation (illuminance was reduced by 50%). On the day of the experiment (before the application of ¹⁴СО₂), several shaded plants were exposed to direct sunlight for 3 min, and some plants grown at full light (light plants) were shaded for 3 min (illuminance of 50%). Unfertilized plants adapted for 5 days to shading showed photosynthesis of 75.9% of control level (full light). If light plants were transferred to shading for 3 min, their photosynthesis decreased to 42.1%. In plants shaded for 5 days and then transferred to full light, photosynthesis in 3 min was 96.3% of control level. At full light, fertilization with nitrate boosted photosynthesis to 132.6% as compared with control material, but photosynthesis decreased to 43.5 and 65.4% of control level in plants shaded for 5 days and those shaded for 3 min, respectively. At the same time, the plants shaded for 5 days and then exposed for 3 min to full light restored photosynthesis to almost control level (95.5%). Analysis of ¹⁴С distribution among the products of 3-min-long photosynthesis showed that, the same as in C₃ plants, a decrease in illuminance (especially a sudden one) in maize reduced the ratio between labeled sucrose and hexoses and elevates incorporation of ¹⁴С into malate, which indicated that its consumption in bundle sheath cells was suppressed. A decrease in the ratio between labeled sucrose and hexoses became more pronounced under the influence of nitrates with this effect also occurring in transport products of photosynthesis (20 cm below ¹⁴С-providing leaf area). In plants fertilized with nitrates, radioactivity of sucrose (% of radioactivity of soluble compounds) decreased in all the types of illumination. When illuminance was suddenly reduced for 3 min, incorporation of ¹⁴С into sucrose was 21.5 against 51.2% in light plants, and radioactivity of aspartate and malate sharply rose to 13.7 and 26.1% (against 2.1 and 8.9% in control material). Incorporation of ¹⁴С into compounds of glycolate pathway was low (less than 2.5%), but it was somewhat greater in nitrate plants. We concluded that the same mechanism of interaction between stomatal apparatus of leaf epidermis, invertase of mesophyll apoplast, and photosynthetic metabolism of carbon with electron flux via electron transport chain in chloroplasts of bundle sheath cells, which governs the rate of photosynthesis and assimilate export from the leaf but is triggered by the extent of consumption in the bundle sheath cells of C₄ acids produced in the mesophyll operates in C₄ plants (the same as in C₃ plants).     

Genetic variation of <Emphasis Type="Italic">Ulva (Enteromorpha) prolifera</Emphasis> (Ulvales,Chlorophyta)—the causative species of the green tides in the Yellow Sea,China

Ulva (Enteromorpha) prolifera, widely distributed from the intertidal to the upper subtidal zones around the world, was the dominant species of the massive green tides in the Yellow Sea in the summers of 2007, 2008, and 2009. However, little is known about its intra-species genetic diversity. In this study, six attached and seven floating U. prolifera samples collected from different sites distributed from the north of the Yellow Sea to the south of the East China Sea were taken in inter-simple sequence repeat (ISSR) analysis. Based on the results of the 90 polymorphic bands from four ISSR primers, the genetic diversity level of the floating samples (H = 0.1663, I = 0.2608) was found to be lower than that of the attached samples (H = 0.2105, I = 0.3346). Unweighted pair-group mean analysis (UPGMA) and principal coordinate analysis (PCoA) suggested that floating U. prolifera samples in the Yellow Sea from 2007 to 2009 had a close genetic relationship, and the floating samples were separated from the attached samples. Genetic differentiation and limited gene flow among attached U. prolifera populations were indicated by analysis of molecular variance.     

Food spectra of the farm juveniles of the Chum Salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> (Salmoniformes,Salmonidae) in the Ryazanovka River (Southern Primor’e)

Qualitative data are presented on the diet of juvenile chum salmon Oncorhynchus keta released from the Ryazanovka experimental fish hatchery farm situated in the south of Primor’e Territory. The stomach fullness, frequency of occurrence of food items, their average weight value, and selectivity are estimated. Food spectra of chum salmon juveniles during downstream migration comprised representatives of 51 taxa of bottom freshwater invertebrates and terrestrial insects. The bulk of the diet consisted of larvae of aquatic insects—chironomids, other Diptera, and mayflies. Their diversity increased with the growth of fry. Fry of chum salmon had a wide spectrum of feeding preference, favorite food items were larvae and pupae of chironomids.     

Analysis of the meiosis in the F_1 hybrids of Longiflorum × Asiatic(LA) of lilies(Lilium) using genomic in situ hybridization

Longiflorum and Asiatic lilies of the genus Lilium of the family Liliaceae are two important groups of modern lily cultivars.One of the main trends of lily breeding is to realize introgression between these groups.With cut style pollination and embryo rescue,distant hybrids between the two groups have been obtained.However,the F1 hybrids are highly sterile or some of them could produce a small number of 2n gametes,and their BC1 progenies are usually triploids.Dutch lily breeders have selected many cultivars...     

Ecological Monitoring of the Water Area of Port Vostochnyi (Wrangel Bay) in the Sea of Japan (1995–2002)

Benthic surveys carried out in 1989 (literature data), 1995, and 2001 (original data) revealed changes in the structure of soft bottom communities of Wrangel Bay (Peter the Great Bay, Sea of Japan), where a deep-water port of Vostochnyi is situated. The functioning of the port has caused no significant environmental pollution by heavy metals, hydrocarbons, and other pollutants. Bottom dredging and the siltation induced by it have drastically lowered the benthos biomass, changed benthic communities, and altered their trophic structure. Sestonophages have disappeared nearly altogether being replaced by detrivores. The ecological state of the benthos is characterized as unstable.     

Accuracy of the StressEraser<Superscript>®</Superscript> in the Detection of Cardiac Rhythms

StressEraser is a commercially marketed biofeedback device designed to enhance heart rate variability. StressEraser makes its internal calculations on beat-to-beat measures of finger pulse intervals. However, the accuracy and precision of StressEraser in quantifying interbeat intervals using finger pulse intervals has not been evaluated against standard laboratory equipment using R-R intervals. Accuracy was assessed by simultaneously recording interbeat intervals using StressEraser and a standard laboratory ECG system. The interbeat intervals were highly correlated between the systems. The average deviation in interbeat interval recordings between the systems was approximately 6 ms. Moreover, correlations approached unity between the systems on estimates of heart period, heart rate, and heart rate variability. Feedback from StressEraser is based on an interbeat time series that provides sufficient information to provide an excellent estimate of the dynamic changes in heart rate and heart rate variability. The slight variations between StressEraser and the laboratory equipment in quantifying heart rate and heart rate variability are due to features related to monitoring heart rate with finger pulse: (1) a lack in precision in the peak of the finger pulse relative to the clearly defined inflection point in the R-wave, and (2) contribution of variations in pulse transit time.     

Monitoring the invasion of the aquatic bug <Emphasis Type="Italic">Trichocorixa verticalis verticalis</Emphasis> (Hemiptera: Corixidae) in the wetlands of Doñana National Park (SW Spain)

We have detected the presence of the North American native corixid Trichocorixa verticalis verticalis (Fieber, 1851) in Doñana wetlands (SW Spain). We have collected data from different research projects done in the area during the period of 2001–2007. We have sampled 134 different sites in Doñana and we found the exotic corixid in 66 occasions. We have found two reproductive populations that might act as sources for the colonization of other waterbodies in the area. When reproduction occurred T. v. verticalis outcompeted native corixids. Its presence out of the waterbodies where we detected reproduction was in small numbers and probably due to vagrant individuals.     

Role of the guanidine group in the N-terminal fragment of PTH(1–11)

A series of PTH hybrids containing a diamine [NH₂(CH₂) _n NH₂; n = 4, 5, 6] in the C-terminal position was synthesized based on the H-Aib-Val-Aib-Glu-Ile-Gln-Leu-Nle-His-Gln-Har-NH₂ (Har = homoarginine) template. The compounds were pharmacologically characterized at PTH1R receptors for agonist activity.