Response of external inspiration to the movements induced by transcutaneous spinal cord stimulation

The dynamic of the parameters of lung ventilation and gas exchange have been studied in 10 young male subjects during involuntary stepping movements induced by transcutaneous spinal cord electrical stimulation applied in the projection of T ₁₁–T ₁₂ vertebrae and during voluntary stepping movements. It has been found that the transcutaneous spinal cord stimulation inducing stepping movements leads to an increase in breathing frequency and a reduction in tidal volume. These effects may be mediated by some neurogenic factors associated with muscular activity during stepping movements, the activation of abdominal expiratory muscles, and the interaction between the stepping pattern and breathing generators.     

Altered excitability of and synaptic effects of motoneurons during the establishing of a locomotor generation in the cat

Research was performed on spinal cats injected with DOPA and decorticate (decerebrated at level A 13) and spinal cats. It was found that formation (activation) of the spinal locomotor generator is accompanied by heightened excitability in the extensor and the reverse trend in the flexor motoneurons, by an increase in the efficacy of recurrent and reciprocal Ia inhibition of -motoneurons, and by a weakening of the influence of Ib afferents and extensor reflex afferents on these same motoneurons. The likely functional role of these changes in tuning of the spinal segmental apparatus in the generation of locomotor rhythm is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 18, No. 5, pp. 679–687, September–October, 1986.     

Effect of cold adaptation of α-and β-adrenergic responses of blood pressure in hindlimb and small intestine in situ and systemic blood pressure in rabbits

Changes in the main parameters of α-and β-adrenergic responses, sensitivity to agonists (EC ₅₀) and maximum response (P _m) of hindlimb and small intestinal blood pressure in situ and systemic blood pressure were studied in rabbits adapted to cold for 1–30 days (daily exposures to ?10°C for 6 h). The responses to phenylephrine, noradrenaline, adrenaline, clonidine (α-agonists), and isopropylnoradrenaline (β-agonist) corresponded to the equation p = (P _m A ⁿ )/(EC ₅₀ ⁿ + A ⁿ ) (1) with n = 1 and n = 2, respectively. Cold adaptation induced reciprocal changes in the response of both EC ₅₀ and P _m to α-agonists and in the response of P _m alone to isopropylnoradrenaline. The significant differences of the parameters from control observed during the first 5 days of adaptation gradually decreased by day 30. After 10 days of adaptation, the efficiency (E = P _m/2EC ₅₀) of response to α-and β-agonists of adrenoceptors significantly increased.     

Isolation and characterization of α-amylase inhibitor from <Emphasis Type="Italic">Leucas aspera</Emphasis> (Willd) Link: α-amylase assay combined with FPLC chromatography for expedited identification

The aim of the present study was to isolate and characterize a proteinaceous α-amylase inhibitor from the whole plant extract of Leucas aspera (Willd) Link. The proteins were further purified by fast and reliable ion-exchange chromatography. A ~ 28 kDa protein from L. aspera inhibited the activity of fungal α-amylase by 90% at 80:1 (inhibitor:enzyme) ratio. The inhibition activity was examined in various α-amylases and its enhanced inhibition activity was witnessed. The activity of the inhibitor on α-amylase was stable and high at pH 6–7 and at temperatures of 30–50 °C. The high-resolution α-amylase inhibition assay/FPLC-MS-SPE platform allowed identification of 28 kDa protein with high purification fold as the α-amylase inhibitor in L. aspera and peptides were matched with highest score of alpha-amylase/trypsin inhibitor of Zea mays. In conclusion, results here obtained suggested that the primary metabolites (proteins) in L. aspera are mainly responsible for its versatile biological and pharmacological activities.     

Antioxidant and α-glucosidase inhibitory phenolic constituents of <Emphasis Type="Italic">Lactuca indica</Emphasis> L.

Lactuca indica L. (Compositae) has been used as a folk medicine for the treatment of intestinal disorders. Phytochemical constituents of L. indica were investigated, and their antioxidant and α-glucosidase inhibitory activities thoroughly studied. A phytochemical investigation of the aerial parts of L. indica resulted in the isolation and identification of eight phenolic compounds. The structures of the compounds were elucidated on the basis of spectroscopic evidence as apigenin, luteolin, isoquercitrin, chlorogenic acid, protocatechuic acid, p-hydroxymethyl benzoic acid, trans-cinnamic acid, and p-coumaric acid. Luteolin, isoquercitrin, chlorogenic acid, and p-hydroxymethyl benzoic acid showed antioxidant activities with IC₅₀ values in the range of 35.5–52.5 μM. In addition, apigenin and luteolin showed α-glucosidase inhibition activities with IC₅₀ values of 96.4 and 100.7 μM, respectively. These findings strongly suggest that L. indica is a potential source of natural antioxidants and/or anti-diabetic agents in pharmaceutical and health functional foods.     

Genomic and functional characterization of coleopteran insect-specific α-amylase inhibitor gene from Amaranthus species

The smallest 32 amino acid α-amylase inhibitor from Amaranthus hypochondriacus (AAI) is reported. The complete gene of pre-protein (AhAI) encoding a 26 amino acid (aa) signal peptide followed by the 43 aa region and the previously identified 32 aa peptide was cloned successfully. Three cysteine residues and one disulfide bond conserved within known α-amylase inhibitors were present in AhAI. Identical genomic and open reading frame was found to be present in close relatives of A. hypochondriacus namely Amaranthus paniculatus, Achyranthes aspera and Celosia argentea. Interestingly, the 3′UTR of AhAI varied in these species. The highest expression of AhAI was observed in A. hypochondriacus inflorescence; however, it was not detected in the seed. We hypothesized that the inhibitor expressed in leaves and inflorescence might be transported to the seeds. Sub-cellular localization studies clearly indicated the involvement of AhAI signal peptide in extracellular secretion. Full length rAhAI showed differential inhibition against α-amylases from human, insects, fungi and bacteria. Particularly, α-amylases from Helicoverpa armigera (Lepidoptera) were not inhibited by AhAI while Tribolium castaneum and Callosobruchus chinensis (Coleoptera) α-amylases were completely inhibited. Molecular docking of AhAI revealed tighter interactions with active site residues of T. castaneum α-amylase compared to C. chinensis α-amylase, which could be the rationale behind the disparity in their IC₅₀. Normal growth, development and adult emergence of C. chinensis were hampered after feeding on rAhAI. Altogether, the ability of AhAI to affect the growth of C. chinensis demonstrated its potential as an efficient bio-control agent, especially against stored grain pests.     

Immunoreactivity of synapses on primary afferent axons and sensory neurons in lamprey spinal cord (<Emphasis Type="Italic">Lampetra fluviatilis</Emphasis>)

The ultrastructure and immunospecificity of synapses on primary afferents and dorsal sensory cells (DCs) were studied in lamprey (Lampetra fluviatilis) spinal cords. Using the postembedding immunogold method with a combination of antibodies—polyclonal antibodies to glutamate and monoclonal antibodies to gamma-aminobutyric acid (GABA)—the presence of GABA-positive on the primary afferent axons and GABA-and glutamate-immunopositive synapses on the DC somatic membranes have been shown. Thus, it is obvious that sensory information in the lamprey is controlled by both presynaptic inhibition via synapses on the primary afferent axons and by direct synaptic influence on the body of the sensory neuron.     

Molecular characterization of α-gliadin gene sequences in Indian wheat cultivars vis-à-vis celiac disease eliciting epitopes

α-Gliadin proteins of the wheat gluten form a multigene family encoded by genomic loci Gli-A2, Gli-B2 and Gli-D2 located on the homoeologous wheat chromosomes 6AS, 6BS, and 6DS, respectively which upon partial digestion elicits celiac disease (CD) in the genetically susceptible individuals. The present investigation was planned to study the variations in the amino acid sequence of the α-gliadin proteins and CD eliciting epitopes in the Indian wheat cultivars. Representative wheat varieties released and cultivated in India during the period 1905–2011 were selected for studying the α-gliadin genes by cloning and sequencing followed by in silico analysis of the gene sequences. A lot of variation for α-gliadin gene sequences especially in T cell stimulatory epitopes glia-α9, glia-α20, glia-α2 and glia-α was observed in different wheat varieties. Modern varieties released during 1971–2011 had higher proportion of intact T-cell stimulatory epitopes. The old wheat varieties released in the period 1905–1970 on the other hand had large proportion of variant epitopes. We identified three wheat varieties namely C591, C273 and K78 having only variant epitopes at Gli-D2 and Gli-B2 and both intact and variant epitopes at Gli-A2. Identification of lower proportion of T-cell stimulatory epitopes in these three varieties is the first step towards developing a wheat variety less immunogenic for celiac disease patients. The gene sequences of the selected varieties have been submitted at NCBI with accession numbers GenBank KJ410473–KJ410488.     

A mutation of casein kinase 2 α4 subunit affects multiple developmental processes in <Emphasis Type="Italic">Arabidopsis</Emphasis>

Key message

Arabidopsis CK2 α4 subunit regulates the primary root and hypocotyl elongation, lateral root formation, cotyledon expansion, rosette leaf initiation and growth, flowering, and anthocyanin biosynthesis.

Abstract

Casein kinase 2 (CK2) is a conserved tetrameric kinase composed of two α and two β subunits. The inhibition of CK2 activity usually results in severe developmental deficiency. Four genes (CKA1–CKA4) encode CK2 α subunit in Arabidopsis. Single mutations of CKA1, CKA2, and CKA3 do not affect the normal growth of Arabidopsis, while the cka1 cka2 cka3 triple mutants are defective in cotyledon and hypocotyl growth, lateral root development, and flowering. The inhibition of CKA4 expression in cka1 cka2 cka3 background further reduces the number of lateral roots and delays the flowering time. Here, we report the characterization of a novel knockout mutant of CKA4, which exhibits various developmental defects including reduced primary root and hypocotyl elongation, increased lateral root density, delayed cotyledon expansion, retarded rosette leaf initiation and growth, and late flowering. The examination of the cellular basis for abnormal root development of this mutant revealed reduced root meristem cells with enhanced RETINOBLASTOMA-RELATED (RBR) expression that promotes cell differentiation in root meristem. Moreover, this cka4-2 mutant accumulates higher anthocyanin in the aerial part and shows an increased expression of anthocyanin biosynthetic genes, suggesting a novel role of CK2 in modulating anthocyanin biosynthesis. In addition, the complementation test using primary root elongation assay as a sample confirms that the changed phenotypes of this cka4-2 mutant are due to the lack of CKA4. Taken together, this study reveals an essential role of CK2 α4 subunit in multiple developmental processes in Arabidopsis.

     

Culture Optimization Strategy for 1-Deoxynojirimycin-producing <Emphasis Type="Italic">Bacillus methylotrophicus</Emphasis> K26 Isolated from Korean Fermented Soybean Paste, <Emphasis Type="Italic">Doenjang</Emphasis>

1-Deoxynojirimycin (1-DNJ) is an α-glucosidase inhibitor that is used for the treatment of type 2 diabetes. In this study, we isolated Bacillus methylotrophicus K26 with α-glucosidase inhibition (AGI) activity from Korean fermented soybean paste (Doenjang) and confirmed that the genome harbored the DNJ biosynthesis genes including gabT1, yktc1, and gutB1 by PCR screening, while 1-DNJ production was confirmed by ultra-performance liquid chromatography–quadrupole time-of-flight–mass spectrometry. To increase 1-DNJ production by B. methylotrophicus K26, culture conditions were optimized with one-factor-ata- time (OFAT) and response surface methodology (RSM) approaches. Screen of 11 carbon and 9 nitrogen sources by the OFAT method identified sucrose and yeast extract as optimal culture components. Sucrose concentration (X₁), yeast extract concentration (X₂), and culture temperature (X₃) were selected as independent variables for central composite design. The coefficient of determination (R²) for the model was 0.927, and the probability value of the regression model was highly significant. RSM predicted the optimal conditions for 1-DNJ production by B. methylotrophicus K26 as sucrose and yeast extract concentrations of 4.61% and 7.03%, respectively, at a temperature of 34°C. Under these conditions, AGI activity was experimentally measured as 89.3%, which was close to the predicted value of 91.9%.     

Biomass allocation patterns in forests growing different climatic zones of China

Key message

Temperature generally explained variation in branch and leaf biomasses, whereas stem and root biomasses–temperature relationships restricted certain age stages may not hold at broader age ranges.

Abstract

In this study, biomass data for alpine temperate Larix forest, alpine Picea-Abies forest, temperate typical deciduous broadleaved forest, temperate Pinus tabulaeformis forest, temperate mixed coniferous-broadleaved forest, montane Populus-Betula deciduous forest, subtropical evergreen broadleaved forest, subtropical montane Cupressus and Sabina forest, subtropical Pinus massoniana forest and subtropical Cunninghamia lanceolata forest were used to examine the effect of temperature on biomass allocations between organs. The data of the ten forests were classified as ≤30, 31–60 and >60 years, to test whether biomass allocations of these age group forests vary systematically in their responses to temperature. With increasing mean annual temperature, branch and leaf biomasses significantly increased in ≤30, 31–60 and >60 years and all age groups; stem biomass significantly increased in ≤30-, 31–60- and >60-year groups, but no significant trend in all age groups; Root biomass significantly increased in 31–60, >60 years and all age groups, but had no response to mean annual temperature in the 30-year group, which suggest that root biomass allocation in response to temperature is dependent upon forest age. We conclude that temperature generally explained variation in branch and leaf biomasses, whereas stem and root biomasses–temperature relationships restricted certain age stages may not hold at broader age ranges.

     

Fish production in estuaries of Primorye

Production of fish communities in 15 different-types estuaries was assessed based on the data collected as a result of 90 surveys (860 seine stations) conducted in Primorye in 2002–2015. The main contribution to production was made by semi-anadromous species (the so-iuy mullet Liza haematocheilus, Far Eastern redfins Tribolodon spp., and Japanese smelt Hypomesus nipponensis, etc.). In both external (EP _ext) and internal (EP _int) polyhaline estuaries (EP) a substantial portion of the production was provided by resident marine species (mainly by the saffron cod Eleginus gracilis, Far Eastern smooth flounder Liopsetta pinnifasciata, and tidepool gunnel Pholis nebulosa) and southern immigrants (flathead grey mullet Mugil cephalus, dotted gizzard shad Konosirus punctatus, Japanese halfbeak Hyporhamphus sajori, and Pacific needlefish Strongylura anastomella). In mesohaline (EM) and oligohaline (EO) estuaries, the proportion of marine residents and southern immigrants was reduced to a minimum, while that of freshwater species (bighead gudgeon Gobio macrocephalus, Prussian carp Carassius gibelio, minnow Phoxinus spp., Amur bitterling Rhodeus sericeus, spiny bitterlings Acanthorhodeus spp., etc.) substantially added to the production of semi-anadromous fish. The mean fish biomass for the vegetation season varied in the 143–1463 mgC/m² range; the mean annual production was 174–4267 mgC/m² and the mean P/B ratio was 0.2–3.2. In 2007, the high annual production in the Artemovka River estuary, 9356 mgC/m², was formed by the juvenile so-iuy mullet of the strong yearclass that hatched in 2006. The lowest mean production and P/B values were typical mainly for the water bodies with a salinity more often close to that of the barrier zones (5–8‰ for α-horohalinicum and 22–26‰ for β-horohalinicum), i.e., for EM and EP _ext. This relationship is explained by the features of the osmotic regulation in fish of various origins and its ontogenetic variations. In particularly, the salinity in EM is more frequently close to the critical salinity (5–8‰, α-horohalinicum); thus, the proportion of juveniles of most of the species in the catches decreases (as their resistance to salinity variations is lower). This results in higher mean values of the specific and absolute production of fish communities in EP _int and EO as compared to those in EM. The conclusion was made that the estimates of fish production in the estuaries of Primorye are similar to those in the well-studied estuaries of the temperate, subtropical, and tropical zones. Moreover, they are comparable to the fish production estimates for mesotrophic and eutrophic lakes in northwestern Russia, substantially lower than those for large lowland rivers, and higher than those for small rivers. The fish production in seas, including the Japan/East Sea (0.20 gC/m²) and, particularly, Peter the Great Bay (0.28 gC/m²), is mostly lower than that in Primorye estuaries.     

Defense Responses and Changes in Symbiotic Gut Microflora in the Colorado Potato Beetle <Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> under the Effect of Endophytic Bacteria from the Genus Bacillus

Phytophagous insects and host plants have a complex of microsymbionts and make up a united co-evolving system with them. Microsymbiotic complexes are actively involved in stress responses of macrosymbionts. We established that a treatment of potato plants with endophytic bacterial strains Bacillus thuringiensis var. thuringiensis-5689, B. th. var. kurstaki-5351, and Bacillus subtilis 26D decreased the survival rate of the plant feeder, Colorado potato beetle Leptinotarsa decemlineata Say. The B. th. strains suppressed phenoloxidase and acetylcholinesterase activities in the beetle hemolymph. An antagonistic relationship was found between endophytic bacteria B. subtilis 26D and beetle symbiotic bacteria from the genera Acinetobacter and Enterobacter, with the former being able to suppress the growth of endophytic colonies. The recombinant B. subtilis strain 26D Cry, containing the B. th. var. kurstaki δ-endotoxin cry1Ia gene, combined the ability of the original B. subtilis 26D strain to suppress the development of beetle symbionts and immune responses with a production of the Cry toxin, thus leading to a high mortality of the phytophage.     

The Effect of Transcutaneous Electrical Spinal Cord Stimulation on the Functional Activity of Spinal Inhibition in the System of Synergistic Muscles of the Lower Leg in Humans

Human Physiology - The effect of 20-minute transcutaneous electrical spinal cord stimulation (tESCS) on the severity of nonreciprocal and recurrent inhibition of spinal α-motoneurons in humans...     

Growth of Ag-nanoparticles in an aqueous solution and their antimicrobial activities against Gram positive,Gram negative bacterial strains and Candida fungus

Silver nanoparticles (AgNPs) were synthesized using Ocimum sanctum (Tulsi) leaves aqueous extract as reducing as well as a capping agent in absence and presence of cetyltrimethylammonium bromide (CTAB). The resulting nanomaterials were characterized by UV–visible spectrophotometer, and transmission electron microscope. The UV–Vis spectroscopy revealed the formation of AgNPs at 400–450 nm. TEM photographs indicate that the truncated triangular silver nanoplates and/or spherical morphology of the AgNPs with an average diameter of 25 nm have been distorted markedly in presence of CTAB. The AgNPs were almost mono disperse in nature. Antimicrobial activities of AgNPs were determined by using two bacteria (Gram positive Staphylococcus aureus MTCC-3160), Gram negative Escherichia coli MTCC-450) and one species of Candida fungus (Candida albicans ATCC 90030) with Kirby-Bauer or disc diffusion method. The zone of inhibition seems extremely good showing a relatively large zone of inhibition in both Staphylococcus aureus, Escherichia coli, and Candida albicans strains.     

Comparison of Alleles at <Emphasis Type="Italic">Gli-2</Emphasis> Loci of Common Wheat by Means of Two-Dimensional Electrophoresis of Gliadin

Electrophoretic mobility (EM) and molecular weight (MW) of some allelic variants of α- and β-gliadins contrlled by Gli-2 loci were compared by means of two-dimensional (APAGE × SDS) electrophoresis. Comparison of α-gliadins of the alleles Gli-A2b and Gli-A2p, of β-gliadins of the Gli-B2b and Gli-B2c, and of β-gliadins of the Gli-D2b, Gli-D2c, Gli-D2j, and Gli-D2r indicated that a gliadin with lower EM had, as a rule, bigger MW which is known to depend on the length of the polyglutamine domain of gliadin of α-type. However, allelic variants of the α-gliadin encoded by Gli-D2b and Gli-D2e differ in EM but not in apparent MW. It might be caused by a substitution of some charged/uncharged aminoacids in the polypeptide of gliadin. Allele Gli-B2o which is very frequent in up-to-date common wheat germplasm originated probably by means of unequal crossingover. Some alleles at Gli-A2 is found to control completely different blocks of gliadins and therefore might come to common wheat from different genotypes of the polymorphic diploid donor of the A genome. The results indicate that the reason of the known more vast polymorphism of gliadins controlled by Gli-2 loci as compared with Gli-1 loci is the considerable difference of the structure, first, of Gli-1 and Gli-2 loci (Gli-2 loci have more expressed genes per locus) and, second, of genes encoding gliadins of α- and γ-types (α-gliadins are shown to contain a long polyglutamine sequences highly variable in their length).     

Crystal and solution structure of the C-terminal part of the <Emphasis Type="Italic">Methanocaldococcus jannaschii</Emphasis> A<Subscript>1</Subscript>A<Subscript>O</Subscript> ATP synthase subunit E revealed by X-ray diffraction and small-angle X-ray scattering

The structure of the C-terminus of subunit E (E_101–206) of Methanocaldococcus jannaschii A-ATP synthase was determined at 4.1 Å. E_101–206 consist of a N-terminal globular domain with three α-helices and four antiparallel β-strands and an α-helix at the very C-terminus. Comparison of M. jannaschii E_101–206 with the C-terminus E_81–198 subunit E from Pyrococcus horikoshii OT3 revealed that the kink in the C-terminal α-helix of E_81–198, involved in dimer formation, is absent in M. jannaschii E_101–206. Whereas a major dimeric surface interface is present between the P. horikoshii E_81–198 molecules in the asymmetric unit, no such interaction could be found in the M. jannaschii E_101–206 molecules. To verify the oligomeric behaviour, the low resolution structure of the recombinant E_85–206 from M. jannaschii was determined using small angle X-ray scattering. Rigid body modeling of two copies of one of the monomer established a fit with a tail to tail arrangement.