Unique molecular architecture of silk fibroin in the waxmoth, Galleria mellonella

Proteins of silk fibers are characterized by reiterations of amino acid repeats. Physical properties of the fiber are determined by the amino acid composition, the complexity of repetitive units, and arrangement of these units into higher order arrays. Except for very short motifs of 6-10 residues, the length of repetitive units and the number of these units concatenated in higher order assemblies vary in all spider and lepidopteran silks analyzed so far. This paper describes an exceptional silk protein represented by the 500-kDa heavy chain fibroin (H-fibroin) of the waxmoth, Galleria mellonella. Its non-repetitive N-terminal (175 residues) and C-terminal (60 residues) parts, the overall gene organization, and the nucleotide sequence around the TATA box show that it is homologous to the H-fibroins of other Lepidoptera. However, over 95% of the protein consists of highly ordered repetitive structures that are unmatched in other species. The repetitive region includes 11 assemblies AB(1)AB(1)AB(1)AB(2)(AB(2))AB(2) of remarkably conserved polypeptide repeats A (63 amino acid residues), B(1) (43 residues), and B(2) (18 residues). The repeats contain a high proportion of Gly (31.6%), Ala (23.8%), Ser (18.1%), and of residues with long hydrophobic side chains (16% for Leu, Ile, and Val combined). The presence of the GLGGLG and SSAASAA(AA) motifs suggests formation of pleated beta-sheets and their stacking into crystallites. Conspicuous conservation of the apolar sequence VIVI followed by DD or ED is interpreted as indicating the importance of hydrophobicity and electrostatic charge in H-fibroin cross-linking. The environment of G. mellonella larvae within bee cultures requires continuous production of silk that must be both strong and elastic. The spectacular arrangement of the repetitive H-fibroin region apparently evolved to meet these requirements.     

Lignin degrading system of white-rot fungi and its exploitation for dye decolorization

With global attention and research now focused on looking for the abatement of pollution, white-rot fungi is one of the hopes of the future. The lignin-degrading ability of these fungi have been the focus of attention for many years and have been exploited for a wide array of human benefits. This review highlights the various enzymes produced by white-rot fungi for lignin degradation, namely laccases, peroxidases, aryl alcohol oxidase, glyoxal oxidase, and pyranose oxidase. Also discussed are the various radicals and low molecular weight compounds that are being produced by white-rot fungi and its role in lignin degradation. A brief summary on the developments in research of decolorization of dyes using white-rot fungi has been made.     

Alterations in Ca2+-channels during the development of diabetic cardiomyopathy

In order to examine the status of Ca²⁺ channels in heart sarcolemma during the development of diabetes, rats were injected intravenously with 65 mg/kg streptozotocin and hearts were removed 1, 3 and 8 weeks later. Crude membranes from the ventricular muscle were prepared and the specific binding of ³H-nitrendipine was studied by employing different concentrations of this Ca ²⁺-antagonist. A significant decrease in both dissociation constant and maximal number of 3H-nitrendipine binding was observed in 3 and 8 weeks diabetic preparations. No such alterations were evident in diabetic brain membranes. Treatment of diabetic animals with insulin prevented the occurrence of these changes in the myocardium. The altered ³H-nitrendipine binding characteristics in diabetic heart membranes may not be due to the high levels of circulating catecholamines in this experimental model because no such changes were seen upon injecting a high dose (40 mg/kg) of isoproterenol in rats for 24 hr. The reduced number of ³H-nitrendipine binding sites may decrease Ca²⁺-influx through voltage sensitive Ca²⁺ channels and partly explain the depressed cardiac contractile force development in chronic diabetes whereas the increased affinity of Ca²⁺ channels may partly explain the increased sensitivity of diabetic heart to Ca ²⁺.     

Survival factor-like activity of small peptides in hybridoma and CHO cells cultures

Synthetic peptides containing three to six amino acid residues were previously shown to improve key parameters of monoclonal antibody-producing mouse hybridoma cultures. The aim of the current work was to investigate whether small peptides also exert analogous beneficial impact on a CHO-K1-derived cell line (XMK-111-10) engineered for production of the human model glycoprotein SEAP (secreted alkaline phosphatase). Similar to hybridoma cultures, growth and SEAP production profiles of CHO XMK-111-10 were modulated by peptides. Both viable cell density and SEAP production were increased by tetraalanine or by a fraction of wheat gluten hydrolysate. Whereas tetraglycine increased the peak viable cell density, the growth-suppressing tripeptide Gly-Lys-Gly significantly boosted SEAP production. All peptide-supplemented cultures showed slight improvement of culture viability during the decline phase of the batch cultures, suggesting a survival factor-like activity of the peptides.     

Genomic evidence for divergence with gene flow in host races of the larch budmoth

Ecological divergence in the face of gene flow has recently become implicated as a potentially important cause of speciation and adaptive radiation. Here, we develop a genomic approach to test for divergent selection in sympatric host races of the larch budmoth Zeiraphera diniana (Lepidoptera: Tortricidae). We analysed hundreds of amplified fragment length polymorphism markers in 92 individuals in sympatric and allopatric populations, and in two backcross broods used to map the markers to individual chromosomes. The results directly confirm the existence of natural hybridization and demonstrate strong heterogeneity between chromosomes in terms of molecular divergence between host races (the average level of divergence was FST = 0.216). However, genomic heterogeneity was not found when we analysed divergence between geographically separated populations of the same host race. We conclude that the variance of the level of sympatric divergence among chromosomes is the footprint of divergent selection acting on a few linkage groups, combined with appreciable gene flow that homogenizes between-race variation at the remaining linkage groups. These results, coupled with other recent multilocus analyses of sister species pairs, demonstrate that selection-driven sympatric phase of genetic divergence in the presence of gene flow is a likely feature of speciation.     

Effects of crowding, isolation, and transfer from isolation to crowding on total ecdysteroid content of eggs in Schistocerca gregaria

We examined the ecdysteroid content of eggs from crowd-reared and solitary-reared desert locusts, Schistocerca gregaria, throughout embryogenesis from the day of egg laying until shortly before hatching on day 14. Depending on the time during incubation, ecdysteroid content in eggs from crowd-reared females was 5-10 times higher than in eggs from solitary-reared females. Our investigation revealed two peaks in ecdysteroid content of eggs from crowd-reared females, a small one at day 3 and a major peak at day 10 of incubation. At days 10, 12 and 14 during incubation of eggs from crowd-reared females, we found a positive correlation between egg mass and ecdysteroid content. There was no difference between eggs from the bottom and the top of individual egg pods, but variation in ecdysteroid content between egg pods from different females was considerable in all treatment groups. A brief crowding of solitary-reared females at the time of egg laying, a treatment that initiates maternally mediated gregarization of the developing offspring, had no effect on the consistently low ecdysteroid content in the eggs. This result rules out the possibility that the crowding experience of females is transmitted to the offspring by variation in the total amount of ecdysteroids in their eggs.     

Redescription of Camallanus hypophthalmichthys Dogel and Akhmerov, 1959 (Nematoda: Camallanidae) and its first record from fishes in China

The nematode Camallanus hypophthalmichthys Dogel and Akhmerov, 1959 is redescribed from specimens collected from the intestine of the bighead carp Aristichthys nobilis, from Liangzihu Lake (Yangtze River basin), Hubei Province, central China. The light and scanning electron microscopical examination made it possible to study in detail the morphology of this so far little-known species and to confirm its validity. The main specific features of C. hypophthalmichthys distinguishing it from the most similar Camallanus spp. is the presence of 3 small caudal processes on the male tail tip, 13-16 longitudinal ridges on the inner surface of the valve of the buccal capsule, and the arrangement of preanal and postanal genital papillae in the male. This finding represents a new host record, the first record of this parasite in the Yangtze River basin, and the first documented record of C. hypophthalmichthys from China. Camallanus hypophthalmichthys is considered a specific intestinal parasite of fishes of the cyprinid Hypophthalmichthyinae.     

A new trichosomoidid from the skin of Sebastes spp. (Pisces) from British Columbia, Canada

A new species of trichosomoidid nematode, Huffinanela canadensis n. sp., is described from the skin of rockfish (Sebastes spp.) from the Pacific Ocean off the coast of Vancouver Island (Clayoquot Sound region), British Columbia, Canada, on the basis of the morphology of the adult worms and their eggs in the host's tissue and the biological characters. The species is characterized mainly by the shape and structure of the fully developed eggs (absence of surface envelope, surface with transverse ridges), by their small size (48-63 x 24-27 microm), and by the site of infection (skin). Besides Huffmanela huffmani, this is the second Huffmanela species in which adult worms are known in addition to the eggs from the host's tissues (most Huffmanela spp. have been described only from their conspicuous eggs occurring in various tissues of fishes). Adults of H. canadensis differ morphologically from those of H. huffmani, mainly in the structure of the male caudal end and in the distinctly elevated anterior vulvar lip of the female.     

Calpain Sensitive Regions in the N-terminal Cytoplasmic Domains of Glycine Transporters GlyT1A and GlyT1B

Glycine transporters are members of the Na⁺/Cl⁻ dependent transporter gene family and play crucial roles in regulating inhibitory as well as excitatory neurotransmission. In this report we show that calcium elevation in spinal cord synaptosomes decreases the levels of glycine transporter, GlyT1, N-terminal immunoreactivity, and that this decrease can be blocked by calpain inhibitor. Sequencing of GST fusion proteins containing the N-terminal domains of GlyT1A and B splice variants cleaved with rat recombinant calpain identified calpain cleavage sites after glycine 17 in GlyT1B and N-terminally of the first conserved arginine residue in both GlyT1A and GlyT1B. Expression in HEK293 cells revealed that truncation of the N-terminus of GlyT1 results in significant inhibition of glycine uptake. A syntaxin1A GST fusion protein was able to pull-down N-terminally deleted GlyT1, indicating that calpain cleavage does not eliminate syntaxin1A binding. These results suggest that calpain cleavage may regulate the transport activity/turnover of GlyT1 in vivo by cleaving its N-terminal domain.     

Localization of the PsbH subunit in photosystem II from the Synechocystis 6803 using the His-tagged Ni-NTA Nanogold labeling

The PsbH protein belongs to a group of small protein subunits of photosystem II (PSII) complex. This protein is predicted to have a single transmembrane helix and it is important for the assembly of the PSII complex as well as for the proper function at the acceptor side of PSII. To identify the location of the PsbH subunit, the PSII complex with His-tagged PsbH protein was isolated from the cyanobacterium Synechocystis sp. PCC 6803 and labeled by Ni(2+)-nitrilo triacetic acid Nanogold. Electron microscopy followed by single particle image analysis identified the location of the labeled His-tagged PsbH protein at the periphery of the dimeric PSII complex. These results indicate that the N terminus of the PsbH protein is located at the stromal surface of the PSII complex and close to the CP47 protein.