Inhibition of Malaria Infection in Transgenic Anopheline Mosquitoes Lacking Salivary Gland Cells

Malaria is an important global public health challenge, and is transmitted by anopheline mosquitoes during blood feeding. Mosquito vector control is one of the most effective methods to control malaria, and population replacement with genetically engineered mosquitoes to block its transmission is expected to become a new vector control strategy. The salivary glands are an effective target tissue for the expression of molecules that kill or inactivate malaria parasites. Moreover, salivary gland cells express a large number of molecules that facilitate blood feeding and parasite transmission to hosts. In the present study, we adapted a functional deficiency system in specific tissues by inducing cell death using the mouse Bcl-2-associated X protein (Bax) to the Asian malaria vector mosquito, Anopheles stephensi. We applied this technique to salivary gland cells, and produced a transgenic strain containing extremely low amounts of saliva. Although probing times for feeding on mice were longer in transgenic mosquitoes than in wild-type mosquitoes, transgenic mosquitoes still successfully ingested blood. Transgenic mosquitoes also exhibited a significant reduction in oocyst formation in the midgut in a rodent malaria model. These results indicate that mosquito saliva plays an important role in malaria infection in the midgut of anopheline mosquitoes. The dysfunction in the salivary glands enabled the inhibition of malaria transmission from hosts to mosquito midguts. Therefore, salivary components have potential in the development of new drugs or genetically engineered mosquitoes for malaria control.     

The amyloid fibrils of the constant domain of immunoglobulin light chain

Light chain-associated (AL) amyloidosis is characterized by dominant fibril deposition of the variable domain (VL) of an immunoglobulin light chain, and thus its constant domain (CL) has been considered not to be amyloidogenic. We examined the in vitro fibril formation of the isolated CL in comparison with β₂-microglobulin (β2-m), an immunoglobulin domain-like amyloidogenic protein responsible for dialysis-related amyloidosis. Two methods useful for β2-m at neutral pH also induced amyloid fibrils of CL, which were monitored by thioflavin-T binding and electron microscopy (EM). These results suggest that CL plays an important role, more than previously assumed, in the development of AL-amyloidosis.     

Benzo[a]pyrene exposed to solar-simulated light inhibits apoptosis and augments carcinogenicity

Polycyclic aromatic hydrocarbons (PAHs) such as benzo[a]pyrene (BaP) are widespread environmental pollutants and several lines of experimental evidence have suggested a role in carcinogenesis. PAHs in the environment are exposed to sunlight and photomodified PAHs have been detected in contaminated sediment and air particulate matter; however, the carcinogenicity of photomodified PAHs is not well understood. In this study, we found that solar-simulated light-irradiated BaP (LBaP) inhibited apoptosis, leading to cancer. LBaP suppressed apoptosis induced by cell detachment and serum depletion in a dose and light-irradiated time-dependent manner. The antiapoptotic effect was related to the production of reactive oxygen species from degraded BaP. The cells that survived apoptosis by LBaP treatment were transformed having the ability to form colonies in soft agar and tumors in nude mice. These capabilities were specific to LBaP, not BaP itself. The results suggested that the carcinogenicity of PAHs may be attributable not only to the genetic damage induced by their metabolites, but also to the antiapoptotic effects of oxidative products on exposure to sunlight.     

Phylogeny of Local Sheep Breeds in East Asia, Focusing on the Bayanbulak Sheep in China and the Sipsu Sheep in Bhutan

The phylogenetic positions of the Bayanbulak sheep in China and the Sipsu sheep in Bhutan in the northern Asian sheep group were determined on the basis of allele frequency data for five informative and polymorphic loci of blood protein and nonproteins, such as transferrin (TF), arylesterase (ES), hemoglobin-β (HB-β), X-protein (XP), and potassium transport (KE), using different electrophoretic and ion-densitometric techniques. Based on Nei’s genetic distance, clustering analysis by the UPGMA method showed that the Bayanbulak sheep is clustered in the northern Asian sheep group. Furthermore, the Bayanbulak sheep belongs to a subgroup containing the Khalkhas and Hu sheep of the Mongolian sheep group, which is distinguished from another subgroup of the small-tailed Han, Tan, Tong, and Wadi sheep. The Bayanbulak sheep was closest to the Hu sheep, despite a morphological difference in the fat deposits. In addition to these findings, the Sipsu sheep was verified to belong to the Baruwal sheep.     

Effects of a triterpenoid saponin on spectral properties of undegraded pea phytochrome

Soyasaponin I, a triterpenoid saponin isolated from etiolated pea (Pisum sativum cv. Alaska) shoots and identified as Pfr killer, was examined for its effects on spectral properties of undegraded pea phytochrome. When soyasaponin I in concentrations of 100 micromolar or lower was added to Pr in the dark, the spectrum of Pr was not significantly affected, whereas in the presence of 120 micromolar or higher concentrations the absorption maximum of Pr shifted from 666 to 658 nanometer with slight decrease of absorbance. After a brief exposure of the mixture to red light, the increase in absorbance at 666 nanometers that occurs in the dark was inhibited at 26 micromolar and higher soyasaponin I concentrations; the maximum effect being reached at about 180 micromolar. The decrease in absorbance at 724 nanometers in the dark after red light irradiation was somewhat inhibited by 60 micromolar and totally prevented by 410 micromolar soyasaponin I. When P₆₅₈ was irradiated with red light in the presence of 220 micromolar or higher soyasaponin I concentrations, a bleached form (P_bl) was produced instead of Pfr. P_bl showed no dark spectral changes, and the phototransformation of P_bl to P₆₅₈ required a significantly high irradiance of far-red light. When the saponin was added to Pfr in the dark, none of the above-described spectral changes occurred, although the same effects were observed after the mixture was exposed briefly to far-red light followed by red light.     

Analysis of exposed cellulose surfaces in pretreated wood biomass using carbohydrate‐binding module (CBM)–cyan fluorescent protein (CFP)

In enzymatic saccharification of lignocellulosics, the access of the enzymes to exposed cellulose surfaces is a key initial step in triggering hydrolysis. However, knowledge of the structure–hydrolyzability relationship of the pretreated biomass is still limited. Here we used fluorescent‐labeled recombinant carbohydrate‐binding modules (CBMs) from Clostridium josui as specific markers for crystalline cellulose (CjCBM3) and non‐crystalline cellulose (CjCBM28) to analyze the complex surfaces of wood tissues pretreated with NaOH, NaOH–Na₂S (kraft pulping), hydrothermolysis, ball‐milling, and organosolvolysis. Japanese cedar wood, one of the most recalcitrant softwood species was selected for the analysis. The binding analysis clarified the linear dependency of the exposure of crystalline and non‐crystalline cellulose surfaces for enzymatic saccharification yield by the organosolv and kraft delignification processes. Ball‐milling for 5–30 min increased saccharification yield up to 77%, but adsorption by the CjCBM–cyan fluorescent proteins (CFPs) was below 5%. Adsorption of CjCBM–CFPs on the hydrothermolysis pulp were less than half of those for organosolvolysis pulp, in coincidence with low saccharification yields. For all the pretreated wood, crystallinity index was not directly correlated with the overall saccharification yield. Fluorescent microscopy revealed that CjCBM3–CFP and CjCBM28–CFP were site‐specifically adsorbed on external fibrous structures and ruptured or distorted fiber surfaces. The assay system with CBM–CFPs is a powerful measure to estimate the initiation sites of hydrolysis and saccharification yields from chemically delignified wood pulps. Biotechnol. Bioeng. 2010; 105: 499–508. © 2009 Wiley Periodicals, Inc.     

Changes of fluorescence spectra of 2'-deoxyadenosine in aqueous solution by radiation

Aqueous solution of 2'-deoxyadenosine (5 X 10(-4) M, buffered at pH 7.0) was irradiated with 60Co gamma-rays under N2, O2, N2O and t-BuOH-N2 atmospheres in order to compare with adenine radiolysis previously reported. By exposure to radiation, the fluorescence was found to increase more markedly than that from adenine under all conditions of radiolysis. This result indicates that not only base moiety but also sugar moiety participate in the formation of highly fluorescent products. In this 2'-deoxyadenosine radiolysis, both OH and e-aq take part in the formation of such products, but OH predominates over over e-aq when both active species are present, as observed in adenine radiolysis.     

Behaviour of phenylalanine ammonia-lyase in carrot cells in suspension cultures

The high PAL activity in carrot cells in suspension culture was found at the linear and early stationary phases, with concomitant increases in phenylal     

Mechanisms of Browning Degradation of d-Fructose in Special Comparison with d-Glucose-Glycine Reaction

Degradation mechanisms of d-fructose by the interaction with amino acids or organic acids in aqueous solution at initial pH 5.5 heated at 100°C were investigated and a substantial difference in mechanisms between fructose degradation and glucose-glycine reaction was presented. d-Fructose browned more intensely than did d-glucose in lower concentration of glycine and/or in earier stage of reaction period. By catalytic action of carboxylate anions without any condensation with amino groups, d-fructose was decomposed to 3-deoxy-d-erythrohexosulose, 5-(hydroxymelhyl)-2-furaldehyde, and a less amount of pyruval-dehyde through caramelization. It was considered that the main path of fructose degradation was 1,2-enolization but 2,3-enolization would also occur to a little extent.