A novel cluster of mariner-like elements belonging to mellifera subfamily from spiders and insects: implications of recent horizontal transfer on the South-West Islands of Japan

Mariner-like elements (MLEs) have been isolated from various eukaryotic genomes and they are divided into 15 subfamilies, including main five subfamilies: mauritiana, cecropia, mellifera/capitata, irritans, and elegans/briggsae. In the present study, MLEs belonging to mellifera subfamily were isolated from various spiders and insects (Hymenoptera and Lepidoptera) inhabiting the South-West Islands of Japan and neighboring regions. MLEs isolated from 15 different species formed a distinct novel cluster in mellifera subfamily. MLEs obtained from three different species [i.e., the bee Amegilla senahai subflavescens (Amsmar1), the wasp Campsomeris sp. (Casmar1), and the swallowtail butterfly Pachliopta aristolochiae (Paamar1)] contained an intact open reading frame that encoded a putative transposase. These transposases exhibited high similarity of 97.9 % among themselves. In case of Casmar1, the presence of an intact ORF was found in high frequencies (i.e., 11 out of 12 clones). In addition, these transposases also showed the presence of a terminal inverted repeat-binding motif, DD(34)D and two highly conserved amino acid motifs, (W/L)(I/L)PHQL and YSP(D/N)L(A/S)P. These two motifs differed from previously known motifs, WVPHEL and YSPDLAP. MLEs isolated from these three different species may have been inserted into their genomes by horizontal transfer. Furthermore, the presence of an intact ORF suggests that they are still active in habitats along these isolated islands.     

Thymoquinone Induces Telomere Shortening,DNA Damage and Apoptosis in Human Glioblastoma Cells

Background

A major concern of cancer chemotherapy is the side effects caused by the non-specific targeting of both normal and cancerous cells by therapeutic drugs. Much emphasis has been placed on discovering new compounds that target tumour cells more efficiently and selectively with minimal toxic effects on normal cells.

Methodology/Principal Findings

The cytotoxic effect of thymoquinone, a component derived from the plant Nigella sativa, was tested on human glioblastoma and normal cells. Our findings demonstrated that glioblastoma cells were more sensitive to thymoquinone-induced antiproliferative effects. Thymoquinone induced DNA damage, cell cycle arrest and apoptosis in the glioblastoma cells. It was also observed that thymoquinone facilitated telomere attrition by inhibiting the activity of telomerase. In addition to these, we investigated the role of DNA-PKcs on thymoquinone mediated changes in telomere length. Telomeres in glioblastoma cells with DNA-PKcs were more sensitive to thymoquinone mediated effects as compared to those cells deficient in DNA-PKcs.

Conclusions/Significance

Our results indicate that thymoquinone induces DNA damage, telomere attrition by inhibiting telomerase and cell death in glioblastoma cells. Telomere shortening was found to be dependent on the status of DNA-PKcs. Collectively, these data suggest that thymoquinone could be useful as a potential chemotherapeutic agent in the management for brain tumours.     

The PCP protein Vangl2 regulates migration of hindbrain motor neurons by acting in floor plate cells,and independently of cilia function

Vangl2, a core component of the Planar Cell Polarity pathway, is necessary for the caudal migration of Facial Branchiomotor (FBM) neurons in the vertebrate hindbrain. Studies in zebrafish suggest that vangl2 functions largely non-cell autonomously to regulate FBM neuron migration out of rhombomere 4 (r4), but the cell-type within which it acts is not known. Here, we demonstrate that vangl2 functions largely in floor plate cells to regulate caudal neuronal migration. Furthermore, FBM neurons fail to migrate caudally in the mouse Gli2 mutant that lacks the floor plate, suggesting an evolutionarily conserved role for this cell type in neuronal migration. Although hindbrain floor plate cilia are disorganized in vangl2 mutant embryos, cilia appear to be dispensable for neuronal migration. Notably, Vangl2 is enriched in the basolateral, but not apical, membranes of floor plate cells. Taken together, our data suggest strongly that Vangl2 regulates FBM neuron migration by acting in floor plate cells, independently of cilia function.     

The Reversed Feto-Maternal Bile Acid Gradient in Intrahepatic Cholestasis of Pregnancy Is Corrected by Ursodeoxycholic Acid

Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific liver disorder associated with an increased risk of adverse fetal outcomes. It is characterised by raised maternal serum bile acids, which are believed to cause the adverse outcomes. ICP is commonly treated with ursodeoxycholic acid (UDCA). This study aimed to determine the fetal and maternal bile acid profiles in normal and ICP pregnancies, and to examine the effect of UDCA treatment. Matched maternal and umbilical cord serum samples were collected from untreated ICP (n = 18), UDCA-treated ICP (n = 46) and uncomplicated pregnancy (n = 15) cases at the time of delivery. Nineteen individual bile acids were measured using HPLC-MS/MS. Maternal and fetal serum bile acids are significantly raised in ICP compared with normal pregnancy (p = <0.0001 and <0.05, respectively), predominantly due to increased levels of conjugated cholic and chenodeoxycholic acid. There are no differences between the umbilical cord artery and cord vein levels of the major bile acid species. The feto-maternal gradient of bile acids is reversed in ICP. Treatment with UDCA significantly reduces serum bile acids in the maternal compartment (p = <0.0001), thereby reducing the feto-maternal transplacental gradient. UDCA-treatment does not cause a clinically important increase in lithocholic acid (LCA) concentrations. ICP is associated with significant quantitative and qualitative changes in the maternal and fetal bile acid pools. Treatment with UDCA reduces the level of bile acids in both compartments and reverses the qualitative changes. We have not found evidence to support the suggestion that UDCA treatment increases fetal LCA concentrations to deleterious levels.     

Staphylococcus aureus produces membrane-derived vesicles that induce host cell death

Gram-negative bacteria produce outer membrane vesicles that play a role in the delivery of virulence factors to host cells. However, little is known about the membrane-derived vesicles (MVs) produced by gram-positive bacteria. The present study examined the production of MVs from Staphylococcus aureus and investigated the delivery of MVs to host cells and subsequent cytotoxicity. Four S. aureus strains tested, two type strains and two clinical isolates, produced spherical nanovesicles during in vitro culture. MVs were also produced during in vivo infection of a clinical S. aureus isolate in a mouse pneumonia model. Proteomic analysis showed that 143 different proteins were identified in the S. aureus-derived MVs. S. aureus MVs were interacted with the plasma membrane of host cells via a cholesterol-rich membrane microdomain and then delivered their component protein A to host cells within 30 min. Intact S. aureus MVs induced apoptosis of HEp-2 cells in a dose-dependent manner, whereas lysed MVs neither delivered their component into the cytosol of host cells nor induced cytotoxicity. In conclusion, this study is the first report that S. aureus MVs are an important vehicle for delivery of bacterial effector molecules to host cells.     

Effects of substrate concentrations on performance of serially connected microbial fuel cells (MFCs) operated in a continuous mode

Stacking of microbial fuel cells (MFC) by connecting multiple small-sized units in a series is used for generating higher power from the MFCs. However, voltage reversal is a critical problem in a serially connected MFC unit. The voltage reversal often occurs when substrate concentration is relatively low in the anodic compartment. Two rectangular individual cells were stacked together in series: MFC1 was fed with 1?g?glucose?L^?1 throughout the experiment while MFC2 was fed with various concentrations of glucose (0.1, 0.2, 0.3, 0.5 and 0.8?g?L^?1). Voltage reversal occurred when the stack configuration was performed using (1?+?0.1)?g?glucose?L^?1. The stacked configurations with (1?+?0.2, 1?+?0.3, 1?+?0.5 and 1?+?0.8)?g?glucose?L^?1 were operated successfully without the voltage reversal. The maximum powers of 1.88, 2.04, 3.6, 2.5 and 2.18?mW were obtained with the stacked configurations of (1?+?0.2), (1?+?0.3), (1?+?0.5), (1?+?0.8) and (1?+?1)?g?glucose?L^?1, respectively. Except in the stacked configuration with (1?+?0.1)?g?glucose?L^?1, the stacked voltages obtained were similar.     

An in silico approach to understand the structure–function properties of a serine protease (Bacifrinase) from Bacillus cereus and experimental evidence to support the interaction of Bacifrinase with fibrinogen and thrombin

Microbial fibrinogenolytic serine proteases find therapeutic applications in the treatment of thrombosis- and hyperfibrinogenemia-associated disorders. However, analysis of structure–function properties of an enzyme is utmost important before its commercial application. In this study, an attempt has been made to understand the structure of a fibrinogenolytic protease enzyme, “Bacifrinase” from Bacillus cereus strain AB01. From the molecular dynamics trajectory analysis, the modelled three-dimensional structure of the protease was found to be stable and the presence of a catalytic triad made up of Asp102, His83 and Ser195 suggests that it is a serine protease. To understand the mechanism of enzyme–substrate and enzyme–inhibitor interactions, the equilibrated protein was docked with human fibrinogen (the physiological substrate of this enzyme), human thrombin and with ten selective protease inhibitors. The Bacifrinase–chymostatin interaction was the strongest among the selected protease inhibitors. The serine protease inhibitor phenyl methane sulphonyl fluoride was found to interact with the Ser134 residue of Bacifrinase. Furthermore, protein–protein docking study revealed the fibrinogenolytic property of Bacifrinase and its interaction with Aα-, Bβ- and Cγ-chains human fibrinogen to a different extent. However, biochemical analysis showed that Bacifrinase did not hydrolyse the γ-chain of fibrinogen. The in silico and spectrofluorometric studies also showed interaction of Bacifrinase with thrombin as well as fibrinogen with a Kd value of 16.5 and .81 nM, respectively. Our findings have shed light on the salient structural features of Bacifrinase and confirm that it is a fibrinogenolytic serine protease.     

Geographical distribution of morphological abnormalities and wing color pattern modifications of the pale grass blue butterfly in northeastern Japan

The pale grass blue butterfly Zizeeria maha has been used as an environmental indicator to evaluate the biological impacts of the Fukushima nuclear accident. A high morphological abnormality rate (AR) of this butterfly was detected in 2011 from radioactively contaminated areas at 37–38°N. However, the geographical AR distribution has not been documented for the entirety of northeastern Japan. Additionally, the geographical distribution of the wing color pattern modification rate (MR) of temperature‐shock type remains undocumented. Here, we collected adult butterflies from many localities in northeastern Japan in 2014 and examined the local AR and MR. Both AR and MR were generally low throughout the 44 local populations surveyed. Latitudinal AR and MR distributions indicated a gap zone at approximately 39°N. The mean AR and the mean MR of the populations south of the gap zone were low (AR = 3.0%, MR = 1.1%), whereas those of the northern populations were relatively high (AR = 10.6%, MR =10.3%). Logistic regression analyses revealed that abnormalities and modifications were associated with temperature‐related variables. We conclude that abnormalities and modifications are generally rare, but that their rates are higher in the northern populations than in the southern ones. These results, along with evidence from other studies, strongly suggest that the high AR detected in 2011 from contaminated areas was induced by anthropogenic radioactive mutagens. This study presents a basic dataset of the current wildlife state of Z. maha in northeastern Japan, which facilitates a future use of this butterfly species as an environmental indicator.     

Relative importance of biotic and abiotic factors affecting bacterial abundance in Lake Biwa: an empirical analysis

To determine the relative importance of factors affecting bacterial abundance in Lake Biwa, correlation and multiple regression analyses were performed with relevant biotic and abiotic variables. Data used in the analyses were collected weekly from April 1997 to June 1998 at a pelagic site in the north basin. The bacterial abundance ranged from 1 to 7 × 10⁶ cells ml⁻¹, and its spatio-temporal pattern was virtually identical to that in previous studies conducted 12–15 years ago. In the surface layer (0–12.5 m), bacterial abundance was significantly correlated with water temperature and with protozoan and metazoan grazers, but not with chlorophyll a and nutrient concentrations. The results suggest that loss factors rather than growth factors are more important in determining bacterial abundance in this lake. However, grazing effects on bacterial abundance differed among zooplankton. Bacterial abundance correlated negatively with phagotrophic nanoflagellates (PNF) and Daphnia, but positively with Eodiaptomus. Thus, PNF and Daphnia act to reduce the bacterial abundance, while Eodiaptomus acts to stimulate. In contrast, these biotic factors did not explain changes in bacterial abundance in the middle (12.5–25 m) and deep (>25 m) layers. Instead, the bacterial abundance in the deep layer was highly correlated with vertical mixing regimes, suggesting that bacterial abundance was directly or indirectly affected by abiotic factors. These results indicate that bacterial abundance in Lake Biwa was regulated by different factors at different depths. Received: February 8, 2000 / Accepted: August 29, 2000