Characterization of tyrosine-rich Antheraea pernyi silk fibroin hydrolysate

Antheraea pernyi silk fibroin (SF) hydrolysate were characterized using UV-VIS spectrometer, amino acid composition and heavy metal contents to explore its potential sources for food or cosmetic additives. The hydrolyzed A. pernyi SF was separated into two parts: (a) SFA, alanine-rich fraction and (b) SFB, tyrosine-rich fraction. SFB exhibited strong absorption peaks at 210 and 280 nm due to the presence of the tyrosine. Heavy metal analysis showed that arsenic and mercury did not detect. Other heavy metals, which includes lead, cadmium, etc., were recorded only a trace amount. Therefore, A. pernyi SF hydrolysate could be safely used as sources of food, cosmetic and pharmaceuticals.     

Docosahexaenoic acid induces the degradation of HPV E6/E7 oncoproteins by activating the ubiquitin–proteasome system

The oncogenic human papillomavirus (HPV) E6/E7 proteins are essential for the onset and maintenance of HPV-associated malignancies. Here, we report that activation of the cellular ubiquitin–proteasome system (UPS) by the omega-3 fatty acid, docosahexaenoic acid (DHA), leads to proteasome-mediated degradation of E6/E7 viral proteins and the induction of apoptosis in HPV-infected cancer cells. The increases in UPS activity and degradation of E6/E7 oncoproteins were associated with DHA-induced overproduction of mitochondrial reactive oxygen species (ROS). Exogenous oxidative stress and pharmacological induction of mitochondrial ROS showed effects similar to those of DHA, and inhibition of ROS production abolished UPS activation, E6/E7 viral protein destabilization, and apoptosis. These findings identify a novel role for DHA in the regulation of UPS and viral proteins, and provide evidence for the use of DHA as a mechanistically unique anticancer agent for the chemoprevention and treatment of HPV-associated tumors.     

DJ-1 null dopaminergic neuronal cells exhibit defects in mitochondrial function and structure: involvement of mitochondrial complex I assembly

DJ-1 is a Parkinson's disease-associated gene whose protein product has a protective role in cellular homeostasis by removing cytosolic reactive oxygen species and maintaining mitochondrial function. However, it is not clear how DJ-1 regulates mitochondrial function and why mitochondrial dysfunction is induced by DJ-1 deficiency. In a previous study we showed that DJ-1 null dopaminergic neuronal cells exhibit defective mitochondrial respiratory chain complex I activity. In the present article we investigated the role of DJ-1 in complex I formation by using blue native-polyacrylamide gel electrophoresis and 2-dimensional gel analysis to assess native complex status. On the basis of these experiments, we concluded that DJ-1 null cells have a defect in the assembly of complex I. Concomitant with abnormal complex I formation, DJ-1 null cells show defective supercomplex formation. It is known that aberrant formation of the supercomplex impairs the flow of electrons through the channels between respiratory chain complexes, resulting in mitochondrial dysfunction. We took two approaches to study these mitochondrial defects. The first approach assessed the structural defect by using both confocal microscopy with MitoTracker staining and electron microscopy. The second approach assessed the functional defect by measuring ATP production, O(2) consumption, and mitochondrial membrane potential. Finally, we showed that the assembly defect as well as the structural and functional abnormalities in DJ-1 null cells could be reversed by adenovirus-mediated overexpression of DJ-1, demonstrating the specificity of DJ-1 on these mitochondrial properties. These mitochondrial defects induced by DJ-1mutation may be a pathological mechanism for the degeneration of dopaminergic neurons in Parkinson's disease.     

Prevalence and distribution of six bee viruses in Korean Apis cerana populations

The prevalence and distribution of six bee viruses was investigated in 527 Apis cerana samples which were collected from five provinces in South Korea. The most prevalent virus, black queen cell virus (BQCV), was present in 75.11% of 446 adult bee samples, followed by sacbrood virus (SBV) in 30.71%. Deformed wing virus (DWV), Kashmir bee virus (KBV), and chronic bee paralysis virus (CBPV) were present at lower levels of 8.07%, 1.56%, and 0.44%, respectively. The most prevalent virus in 81 larvae samples was SBV, with an incidence of 60.49%, followed by BQCV in 48.14%, DWV in 6.17%, and KBV in 1.23% of samples. CBPV infection was not detected in larvae samples, and acute bee paralysis virus (ABPV) was not present in both larvae and adult bee. Simultaneous infections with up to four viruses were also identified. Of these, infections with SBV and BQCV were most frequent in 25.61% of samples. The distribution of these viruses varied considerably throughout the geographic regions investigated. The three provinces of Gyeongbuk, Jeonnam, and Chungbuk had the highest frequency of bee viruses.     

Polycation-mediated enhancement of retroviral transduction efficiency depends on target cell types and pseudotyped Env proteins: implication for gene transfer into neural stem cells

Polycations such as polybrene (PB) are routinely used for most retroviral vector-mediated gene transfer studies because they can increase the infectivity of retroviruses. However, it was not systematically determined if addition of the polycation is an essential prerequisite for all retroviral transductions. To test this, we measured the effects of the polycation on transduction efficiency using various combinations of target cells and pseudotyped viral envelope (Env) proteins. Here, we show polycations do not always increase retroviral transduction efficiency and that their enhancing effect depends on both the type of target cells and Env proteins. The findings presented here also suggest that high transduction rates can be achieved in primary neural stem cells in vitro and in vivo by choosing an appropriate Env protein for pseudotyping without using polycations which are potentially toxic to primary cells and may change the intrinsic characteristics of cells.     

Disulfide bond as a structural determinant of prion protein membrane insertion

Conversion of the normal soluble form of prion protein, PrP (PrP^C), to proteinase K-resistant form (PrP^Sc) is a common molecular etiology of prion diseases. Proteinase K-resistance is attributed to a drastic conformational change from α-helix to β-sheet and subsequent fibril formation. Compelling evidence suggests that membranes play a role in the conformational conversion of PrP. However, biophysical mechanisms underlying the conformational changes of PrP and membrane binding are still elusive. Recently, we demonstrated that the putative transmembrane domain (TMD; residues 111–135) of Syrian hamster PrP penetrates into the membrane upon the reduction of the conserved disulfide bond of PrP. To understand the mechanism underlying the membrane insertion of the TMD, here we explored changes in conformation and membrane binding abilities of PrP using wild type and cysteine-free mutant. We show that the reduction of the disulfide bond of PrP removes motional restriction of the TMD, which might, in turn, expose the TMD into solvent. The released TMD then penetrates into the membrane. We suggest that the disulfide bond regulates the membrane binding mode of PrP by controlling the motional freedom of the TMD.     

Reproductive morphology ofMegaleranthis saniculifolia Ohwi (Ranunculaceae) and its systematic implications

To confirm the taxonomic treatment ofMegaleranthis saniculifolia Ohwi, an endemic genus and species in Korea, we compared its reproductive morphological characteristics with those ofTrollius and other genera within the Ranunculaceae. Although its external morphology might suggest thatMegaleranthis differs fromTrollius, Calathodes, and etc., we found no distinctly different features in this genus. Likewise, previous studies of their pollen structures, chromosome data, and petal morphology have indicated no differences betweenMegaleranthis andTrollius. In fact, related genera share similar characteristics, such as a tetrasporangia anther, glandular tapetum, simultaneous cytokinesis, an anatropous and bitegmic ovule, embryo sac formation of thePolygonum type, exarillate and copious albuminous seed, and several apocarps. Although the unique feature of having both tenuinucellate and crassinucellate ovules simultaneously may initially seem particular toMegaleranthis, it is present in other genera of the same family. Therefore, based on this evidence of reproductive morphology and other information, we suggest thatM. saniculifolia is closely related toTrollius, and should be included within that genus, i.e., asT. chosenensis Ohwi. Nevertheless, we have tentatively placedMegaleranthis within its own monotypic and endemic genus until definitive data become available.     

Prevalence of antibodies in response to <Emphasis Type="Italic">Legionella</Emphasis> species,analysis of a healthy population from Jeollanam-do Province,Korea

Seroepidemological investigation of antibodies to Legionella species in 500 healthy individuals from a single geographical location in Korea was conducted by indirect fluorescent antibody assay (IFA). Considering an antibody titer of > or =1:128 as positive reaction, 15.2% of total sera were positive. In males and females older than 40 years old, levels of IgM and IgG were 1.2% and 14%, respectively. The sera with antibody titers of > or =1:128 to Legionella species accounted for 85 sera, and 9 sera of these were reacted to more than one Legionella species. Reactivity to L. bozemanii, L. micdadei, L. longbeachae, L. pneumophila sg 6, and L. gormanii were 32.9%, 20%, 15%, 10.6%, and 8%, respectively. However, L. pneumophila sg 1, sg 2, and sg 3 did not react to any sera. Serological analysis revealed that the level of antibody in response to L. bozemanii was more prevalent than L. pneumophila. Our results suggest that the antibodies of non-L. pneumophila species, such as L. bozemanii, may be highly prevalent in healthy population within Korea. Although conclusions based on the findings of this study must be cautiously considered given that the population sampled were sourced from a single province, we have added to the knowledge base of serodiagnosis of infections due to non-L. pneumophila species in Korea.