Challenge of studies on the development of new Zn complexes (Zn(opt)₂) to treat diabetes mellitus

The number of worldwide patients suffering from diabetes mellitus (DM) is forecasted to increase over time. The development of compounds without severe side effects for type 2 DM is required not only to treat DM but also to improve the quality of life (QOL) of patients. In this paper, we have described the synthesis of novel first transition metal complexes with S2O2 coordination mode and discussed their anti-diabetic activities. Di(1-oxy-2-pyridinethiolato)Zn complex (Zn(opt)2) with Zn(S2O2) coordination mode displayed higher insulin mimetic with anti-diabetic activity, compared to the ZnCl2 or clinically used medicine (pioglitazone). In addition, Zn(opt)2 improved the insulin and adiponectine levels in the plasma. The gastrointestinal absorption of the Zn complex was found to be higher than that of ZnCl(2). Based on these results, we propose that the Zn(opt)2 complex with Zn(S2O2) coordination mode is a novel candidate for the treatment of type 2 DM; through oral administration.     

Can deliberately incomplete gene sample augmentation improve a phylogeny estimate for the advanced moths and butterflies (Hexapoda: Lepidoptera)?

This paper addresses the question of whether one can economically improve the robustness of a molecular phylogeny estimate by increasing gene sampling in only a subset of taxa, without having the analysis invalidated by artifacts arising from large blocks of missing data. Our case study stems from an ongoing effort to resolve poorly understood deeper relationships in the large clade Ditrysia ( > 150,000 species) of the insect order Lepidoptera (butterflies and moths). Seeking to remedy the overall weak support for deeper divergences in an initial study based on five nuclear genes (6.6 kb) in 123 exemplars, we nearly tripled the total gene sample (to 26 genes, 18.4 kb) but only in a third (41) of the taxa. The resulting partially augmented data matrix (45% intentionally missing data) consistently increased bootstrap support for groupings previously identified in the five-gene (nearly) complete matrix, while introducing no contradictory groupings of the kind that missing data have been predicted to produce. Our results add to growing evidence that data sets differing substantially in gene and taxon sampling can often be safely and profitably combined. The strongest overall support for nodes above the family level came from including all nucleotide changes, while partitioning sites into sets undergoing mostly nonsynonymous versus mostly synonymous change. In contrast, support for the deepest node for which any persuasive molecular evidence has yet emerged (78-85% bootstrap) was weak or nonexistent unless synonymous change was entirely excluded, a result plausibly attributed to compositional heterogeneity. This node (Gelechioidea + Apoditrysia), tentatively proposed by previous authors on the basis of four morphological synapomorphies, is the first major subset of ditrysian superfamilies to receive strong statistical support in any phylogenetic study. A "more-genes-only" data set (41 taxa×26 genes) also gave strong signal for a second deep grouping (Macrolepidoptera) that was obscured, but not strongly contradicted, in more taxon-rich analyses.     

Overexpression of Rhodobacter sphaeroides PufX-bearing maltose-binding protein and its effect on the stability of reconstituted light-harvesting core antenna complex

The PufX protein, encoded by the pufX gene of Rhodobacter sphaeroides, plays a key role in the organization and function of the core antenna (LH1)-reaction centre (RC) complex, which collects photons and triggers primary photochemical reactions. We synthesized a PufX/maltose-binding protein (MBP) fusion protein to study the effect of the PufX protein on the reconstitution of B820 subunit-type and LH1-type complexes. The fusion protein was synthesized using an Escherichia coli expression system and purified by affinity chromatography. Reconstitution experiments demonstrated that the MBP-PufX protein destabilizes the subunit-type complex (20°C), consistent with previous reports. Interestingly, however, the preformed LH1-type complex was stable in the presence of MBP-PufX. The MBP-PufX protein did not influence the preformed LH1-type complexes (4°C). The LH1-type complex containing MBP-PufX showed a unique temperature-dependent structural transformation that was irreversible. The predominant form of the complex at 4°C was the LH1-type. When shifted to 20°C, subunit-type complexes became predominant. Upon subsequent cooling back to 4°C, instead of re-forming the LH1-type complexes, the predominant form remained the subunit-type complexes. In contrast, reversible transformation of LH1 (4°C) and subunit-type complexes (20°C) occurs in the absence of PufX. These results are consistent with the suggestion that MBP-PufX interacts with the LH1α- polypeptide in the subunit (α/β)-type complex (at 20°C), preventing oligomerization of the subunit to form LH1-type complexes.     

Differences in the Head Movement During Baseball Batting between Skilled Players and Novices

ABSTRACT: Nakata, H, Miura, A, Yoshie, M, and Kudo, K. Differences in the head movement during baseball batting between skilled players and novices. J Strength Cond Res 26(10): 2632-2640, 2012-We investigated the pattern of head movement during baseball batting in 8 skilled players and 9 unskilled novices, using a high-speed video camera. The 2 directions of head movement were analyzed as an X-axis (from the home plate to the pitcher's plate) and Z-axis (vertical downward). On the X-axis, peak latency, peak value, the distance from the peak to the value at bat-ball impact, and data variability were compared between the 2 groups. On the Z-axis, peak latency, downward distance, and data variability were analyzed. Peak latency on the X-axis occurred significantly earlier in baseball players than in novices (p < 0.001), and the difference between the minimum peak and impact was significantly larger in the players (p < 0.05). The variability in peak latency on the X-axis was significantly larger in the novices (p < 0.05). The variability in peak value on the Z-axis was also significantly larger in the novices (p < 0.05). Our findings showed that the significant differences in head movement between the 2 groups should help baseball players, beginners, coaches, and strength and conditioning professionals to improve performance, be effectively applied to actual practice, and enhance coaching for batting.     

Construction of an SSR and RAD-Marker Based Molecular Linkage Map of Vigna vexillata (L.) A. Rich

Vigna vexillata (L.) A. Rich. (tuber cowpea) is an underutilized crop for consuming its tuber and mature seeds. Wild form of V. vexillata is a pan-tropical perennial herbaceous plant which has been used by local people as a food. Wild V. vexillata has also been considered as useful gene(s) source for V. unguiculata (cowpea), since it was reported to have various resistance gene(s) for insects and diseases of cowpea. To exploit the potential of V. vexillata, an SSR-based linkage map of V. vexillata was developed. A total of 874 SSR markers successfully amplified single DNA fragment in V. vexillata among 1,336 SSR markers developed from Vigna angularis (azuki bean), V. unguiculata and Phaseolus vulgaris (common bean). An F₂ population of 300 plants derived from a cross between salt resistant (V1) and susceptible (V5) accessions was used for mapping. A genetic linkage map was constructed using 82 polymorphic SSR markers loci, which could be assigned to 11 linkage groups spanning 511.5 cM in length with a mean distance of 7.2 cM between adjacent markers. To develop higher density molecular linkage map and to confirm SSR markers position in a linkage map, RAD markers were developed and a combined SSR and RAD markers linkage map of V. vexillata was constructed. A total of 559 (84 SSR and 475 RAD) markers loci could be assigned to 11 linkage groups spanning 973.9 cM in length with a mean distance of 1.8 cM between adjacent markers. Linkage and genetic position of all SSR markers in an SSR linkage map were confirmed. When an SSR genetic linkage map of V. vexillata was compared with those of V. radiata and V. unguiculata, it was suggested that the structure of V. vexillata chromosome was considerably differentiated. This map is the first SSR and RAD marker-based V. vexillata linkage map which can be used for the mapping of useful traits.     

Neurofibromatosis 2 tumor suppressor, the gene induced by valproic acid, mediates neurite outgrowth through interaction with paxillin

Valproic acid (VPA), the drug for bipolar disorder and epilepsy, has a potent ability to induce neuronal differentiation, yet comparatively little is presently known about the underlying mechanism. We previously demonstrated that c-Jun N-terminal kinase (JNK) phosphorylation of the focal adhesion protein paxillin mediates differentiation in N1E-115 neuroblastoma cells. Here, we show that VPA up-regulates the neurofibromatosis type 2 (NF2) tumor suppressor, merlin, to regulate neurite outgrowth through the interaction with paxillin. The inhibition of merlin function by its knockdown or expression of merlin harboring the Gln-538-to-Pro mutation, a naturally occurring NF2 missense mutation deficient in linking merlin to the actin cytoskeleton, decreases VPA-induced neurite outgrowth. Importantly, the expression of merlin by itself is not sufficient to induce neurite outgrowth, which requires co-expression with paxillin, the binding partner of merlin. In fact, the missense mutation Trp-60-to-Cys or Phe-62-to-Ser, that is deficient in binding to paxillin, reduces neurite outgrowth induced by VPA. In addition, co-expression of a paxillin construct harboring the mutation at the JNK phosphorylation site with merlin results in blunted induction of the outgrowth. We also find that the first LIM domain of paxillin is a major binding region with merlin and that expression of the isolated first LIM domain blocks the effects of VPA. Furthermore, similar findings that merlin regulates neurite outgrowth through the interaction with paxillin have been observed in several kinds of neuronal cells. These results suggest that merlin is an as yet unknown regulator of neurite outgrowth through the interaction with paxillin, providing a possibly common mechanism regulating neurite formation.     

Morphometric characteristics and reproductive capacity of nuclear transplants derived from embryonic cells of loach, Misgurnus anguillicaudatus

Nuclear transplants of loach were produced by transplantating blastula cell nuclei into nonenucleated unfertilized eggs, using recipient eggs and donor cells distinguished by different polymorphic microsatellites. Of the total of 2,847 operated eggs, 143 hatched and 119 developed to the feeding larval stage. For 15 nuclear transplants (i.e., 11.1-year-old fish and 4.2-year-old fish) that survived up to the adulthood, DNA analysis and karyotype analysis were performed. Results showed that, of the 15 fish, 14 had only a nucleus derived from the donor; moreover, 12 were diploids, 1 was a triploid, 1 was a tetraploid, and 1 was a diploid-tetraploid mosaic with both donor and recipient nuclei. For the 12 fish with only a 2n donor nucleus, morphometric analysis was performed, and two female fish and two male fish were mated with normal fish. The fish with only a 2n donor nucleus were determined to be morphometrically identical to normal fish: they had normal gametogenesis and were able to reproduce. Currently, nuclear transplantation technology is beginning to be adopted in fisheries. Biological information on nuclear transplants obtained in this study can be used in the development of nuclear transplantation technology.     

Anthropogenic noise changes arthropod abundances

Anthropogenic noise is a widespread and growing form of sensory pollution associated with the expansion of human infrastructure. One specific source of constant and intense noise is that produced by compressors used for the extraction and transportation of natural gas. Terrestrial arthropods play a central role in many ecosystems, and given that numerous species rely upon airborne sounds and substrate‐borne vibrations in their life histories, we predicted that increased background sound levels or the presence of compressor noise would influence their distributions. In the second largest natural gas field in the United States (San Juan Basin, New Mexico, USA), we assessed differences in the abundances of terrestrial arthropod families and community structure as a function of compressor noise and background sound level. Using pitfall traps, we simultaneously sampled five sites adjacent to well pads that possessed operating compressors, and five alternate, quieter well pad sites that lacked compressors, but were otherwise similar. We found a negative association between sites with compressor noise or higher levels of background sound and the abundance of five arthropod families and one genus, a positive relationship between loud sites and the abundance of one family, and no relationship between noise level or compressor presence and abundance for six families and two genera. Despite these changes, we found no evidence of community turnover as a function of background sound level or site type (compressor and noncompressor). Our results indicate that anthropogenic noise differentially affects the abundances of some arthropod families. These preliminary findings point to a need to determine the direct and indirect mechanisms driving these observed responses. Given the diverse and important ecological functions provided by arthropods, changes in abundances could have ecological implications. Therefore, we recommend the consideration of arthropods in the environmental assessment of noise‐producing infrastructure.     

Metabolic switching of astringent and beneficial triterpenoid saponins in soybean is achieved by a loss‐of‐function mutation in cytochrome P450 72A69

Triterpenoid saponins are major components of secondary metabolites in soybean seeds and are divided into two groups: group A saponins, and 2,3‐dihydro‐2,5‐dihydroxy‐6‐methyl‐4H‐pyran‐4‐one (DDMP) saponins. The aglycone moiety of group A saponins consists of soyasapogenol A (SA), which is an oxidized β‐amyrin product, and the aglycone moiety of the DDMP saponins consists of soyasapogenol B (SB). Group A saponins produce a bitter and astringent aftertaste in soy products, whereas DDMP saponins have known health benefits for humans. We completed map‐based cloning and characterization of the gene Sg‐5, which is responsible for SA biosynthesis. The naturally occurring sg‐5 mutant lacks group A saponins and has a loss‐of‐function mutation (L164*) in Glyma15g39090, which encodes the cytochrome P450 enzyme, CYP72A69. An enzyme assay indicated the hydroxylase activity of recombinant CYP72A69 against SB, which also suggested the production of SA. Additionally, induced Glyma15g39090 mutants (R44* or S348P) lacked group A saponins similar to the sg‐5 mutant, indicating that Glyma15g39090 corresponds to Sg‐5. Endogenous levels of DDMP saponins were higher in the sg‐5 mutant than in the wild‐type lines due to the loss of the enzyme activity that converts SB to SA. Interestingly, the genomes of palaeopolyploid soybean and the closely related common bean carry multiple Sg‐5 paralogs in a genomic region syntenic to the soybean Sg‐5 region. However, SA did not accumulate in common bean samples, suggesting that Sg‐5 activity evolved after gene duplication event(s). Our results demonstrate that metabolic switching of undesirable saponins with beneficial saponins can be achieved in soybean by disabling Sg‐5.     

O2 binding to human serum albumin incorporating iron porphyrin with a covalently linked methyl-L-histidine isomer

We describe the significant difference in the O2 binding affinities of human serum albumin (HSA) incorporating 5,10,15,20-tetrakis{alpha,alpha,alpha,alpha- o-(1'-methylcyclohexanamido)phenyl}porphinatoiron(II) with a covalently linked 1-methyl-L-histidine or 3-methyl-L-histidine [HSA-FeP(1-MHis), HSA-FeP(3-MHis)]. The HSA-FeP(3-MHis) showed an extraordinarily high O2 binding affinity ( P1/2 = 0.2 Torr, 25 degrees C, pH 7.4), which is close to those of relaxed-state hemoglobin and myoglobin. However, replacement of the 3-methyl-L-histidine moiety in FeP(3-MHis) by 1-methyl-L-histidine caused a 35-fold reduction in O2 affinity; the P 1/2 value of HSA-FeP(1-MHis) (22 Torr, 37 degrees C, pH 7.4) is almost identical to that of human red blood cells. Results of kinetic studies indicate that the low O2 binding affinity of FeP(1-MHis) is predominantly manifested in the high O2 dissociation rate constant. In a toluene solution, an identical relationship in the O2 binding property was similarly observed for FeP(1-MHis) and FeP(3-MHis). The axial Fe-N(1-MHis) coordination might be restrained by steric interaction between the 4-methylene group of the histidine and the porphyrin plane.