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A potentially novel actinobacterium isolated from forest soil, Streptomyces sp. KSF103 was evaluated for its insecticidal effect against several mosquito species namely Aedes aegypti, Aedes albopictus, Anopheles cracens and Culex quinquefasciatus. Mosquito larvae and adults were exposed to various concentrations of the ethyl acetate (EA) extract for 24 h. Considerable mortality was evident after the EA extract treatment for all four important vector mosquitoes. Larvicidal activity of the EA extract resulted in LC50 at 0.045 mg/mL and LC90 at 0.080 mg/mL for Ae. aegypti; LC50 at 0.060 mg/mL and LC90 at 0.247 mg/mL for Ae. albopictus; LC50 at 2.141 mg/mL and LC90 at 6.345 mg/mL for An. cracens; and LC50 at 0.272 mg/mL and LC90 at 0.980 mg/mL for Cx. quinquefasciatus. In adulticidal tests, the EA extract was the most toxic to Ae. albopictus adults (LD50 = 2.445 mg/mL; LD90 = 20.004 mg/mL), followed by An. cracens (LD50 = 5.121 mg/mL; LD90 = 147.854 mg/mL) and then Ae. aegypti (LD50 = 28.873 mg/mL; LD90 = 274.823 mg/mL). Additionally, the EA extract exhibited ovicidal activity against Ae. aegypti (LC50 = 0.715 mg/mL; LC90 = 6.956 mg/mL), Ae. albopictus (LC50 = 0.715 mg/mL; LC90 = 6.956 mg/mL), and An. cracens (LC50 = 0.715 mg/mL; LC90 = 6.956 mg/mL), evaluated up to 168 h post-treatment. It displayed no toxicity on the freshwater microalga Chlorella sp. Beijerinck UMACC 313, marine microalga Chlorella sp. Beijerinck UMACC 258 and the ant Odontoponera denticulata. In conclusion, the EA extract showed promising larvicidal, adulticidal and ovicidal activity against Ae. aegypti, Ae. albopictus, An. cracens, and Cx. quinquefasciatus (larvae only). The results suggest that the EA extract of Streptomyces sp. KSF103 has the potential to be used as an environmental-friendly approach in mosquito control. The current study would serve as an initial step toward complementing microbe-based bioinsecticides for synthetic insecticides against medically important mosquitoes.

Co–Fe bimetallic nanoparticles-affixed polyvinylidene fluoride-co-hexafluoropropylene (PVdF-HFP) nanofiber membrane is fabricated using the electrospinning and chemical reduction techniques. The semicrystalline polymeric backbone decorated with the highly crystalline Co–Fe bimetallic nanoparticles enunciates the mechanical integrity, while the incessant and swift electron mobility is articulated with the consistent dissemination of bimetallic nanoparticles on the intersected and multi-layered polymeric nanofibers. The diffusion and adsorption of glucose are expedited in the extended cavities and porosities of as-formulated polymeric nanofibers, maximizing the glucose utilization efficacy, while the uniformly implanted Co4+/Fe3+ active centers on PVdF-HFP nanofibers maximize the electrocatalytic activity toward glucose oxidation under alkaline regimes. Thus, the combinative sorts including nanofiber and nanocomposite strategies of PVdF-HFP/Co–Fe membrane assimilate the enzyme-less electrochemical glucose detection concerts of high sensitivity (375.01 μA mM−1 cm−2), low limit of detection (0.65 μm), and wide linear range (0.001 to 8 mM), outfitting the erstwhile enzyme-less glucose detection reports. Additionally, the endowments of high selectivity and real sample glucose-sensing analyses of PVdF-HFP/Co–Fe along with the binder-less and free-standing characteristics construct the state-of-the-art paradigm for the evolution of affordable enzyme-less electrochemical glucose sensors.

Algal green energy has emerged as an alternative to conventional energy production using fossil fuels. Microbial fuel cells (MFCs), photosynthetic microbial fuel cells (PMFCs) and biophotovoltaic (BPV) platforms have been developed to utilize microalgae for bioelectricity generation, wastewater treatment and biomass production. There remains a lack of research on marine microalgae in these systems, so to the best of our knowledge, all information on their integration in these systems have been gathered in this review, and are used to compare with the interesting studies on freshwater microalgae. The performance of the systems is extremely reliant on the microalgae species and/or microbial community used, the size of the bio-electrochemical cell, and electrode material and distance used. The mean was calculated for each system, PMFC has the highest average maximum power density of 344 mW/m(2), followed by MFC (179 mW/m(2)) and BPV (58.9 mW/m(2)). In addition, the advantages and disadvantages of each system are highlighted. Although all three systems face the issue of low power outputs, the integration of a suitable energy harvester could potentially increase power efficiency and make them applicable for lower power applications.

A facile chemical reduction approach is adopted for the synthesis of iron tungstate (FeWO4)/ceria (CeO2)–decorated reduced graphene oxide (rGO) nanocomposite. Surface morphological studies of rGO/FeWO4/CeO2 composite reveal the formation of hierarchical FeWO4 flower–like microstructures on rGO sheets, in which the CeO2 nanoparticles are decorated over the FeWO4 microstructures. The distinct anodic peaks observed for the cyclic voltammograms of studied electrodes under light/dark regimes validate the electroactive proteins present in the microalgae. With the cumulative endeavors of three-dimensional FeWO4 microstructures, phase effect between rGO sheet and FeWO4/CeO2, highly exposed surface area, and light harvesting property of CeO2 nanoparticles, the relevant rGO/FeWO4/CeO2 nanocomposite demonstrates high power and stable biophotovoltaic energy generation compared with those of previous reports. Thus, these findings construct a distinct horizon to tailor a ternary nanocomposite with high electrochemical activity for the construction of cost-efficient and environmentally benign fuel cells.

The introduction of second language (L2) education in kindergartens is ubiquitous in many places globally; nevertheless, research in these settings is scarce compared with that on older learners. L2 vocabulary development is especially germane to these very young learners, rendering this a research-worthy topic. The present study examined the effects of researcher-designed explicit vocabulary instruction compared with implicit instruction on English-as-a-second-language participants’ (N = 157) gains in not only the target vocabulary items, but also general vocabulary as well as phonological awareness. Statistically significant differences were found in all vocabulary tasks and the phonemic awareness task with small to large effect sizes. These showed that, in addition to the target vocabulary, the participants receiving explicit vocabulary instruction also had greater gains in receptive and expressive general vocabulary and phonemic awareness. The article culminates in delineating the children’s differential achievements, followed by a brief discussion of the limitations and implications.

Funder: Higher Institution Centre of Excellence (HICoE) Fund, Ministry of Education: Air ocean and land interaction; Grant(s): IOES-2014F, The biophotovoltaic cell (BPV) is deemed to be a potent green energy device as it demonstrates the generation of renewable energy from microalgae; however, inadequate electron generation from microalgae is a significant impediment for functional employment of these cells. The photosynthetic process is not only affected by the temperature, CO2 concentration and light intensity but also the spectrum of light. Thus, a detailed understanding of the influences of light spectrum is essential. Accordingly, we developed spectrally optimized light using programmable LED arrays (PLA)s to study the effect on algae growth and bioelectricity generation. Chlorella is a green microalga and contains chlorophyll-a (chl-a), which is the major light harvesting pigment that absorbs light in the blue and red spectrum. In this study, Chlorella is grown under a PLA which can optimally simulate the absorption spectrum of the pigments in Chlorella. This experiment investigated the growth, photosynthetic performance and bioelectricity generation of Chlorella when exposed to an optimally-tuned light spectrum. The algal BPV performed better under PLA with a peak power output of 0.581 mW m−2 for immobilized BPV device on day 8, which is an increase of 188% compared to operation under a conventional white LED light source. The photosynthetic performance, as measured using pulse amplitude modulation (PAM) fluorometry, showed that the optimized spectrum from the PLA gave an increase of 72% in the rETRmax value (190.5 μmol electrons m−2 s−1), compared with the conventional white light source. Highest algal biomass (1100 mg L−1) was achieved in the immobilized system on day eight, which translates to a carbon fixation of 550 mg carbon L−1. When artificial light is used for the BPV system, it should be optimized with the light spectrum and intensity best suited to the absorption capability of the pigments in the cells. Optimum artificial light source with algal BPV device can be integrated into a power management system for low power application (eg. environment sensor for indoor agriculture system).

The exploitation of renewable energy sources for delivering carbon neutral or carbon negative solutions has become challenging in the current era because conventional fuel sources are of finite origins. Algae are being used in the development of biophotovoltaic (BPV) platforms which are used to harvest solar energy for bioelectricity generation. Fast-growing algae have a high potential for converting CO2 from the atmosphere into biomass and valuable products. In photosynthesis light-driven splitting of water occurs, releasing a pair of electrons and generating O2. The electrons can be harvested and converted to bioelectricity. In this study, algal biofilms of a tropical cyanobacterial strain Synechococcus elongatus (UMACC 105) were formed on two types of electrodes, indium tin oxide (ITO) and reduced graphene oxide (rGO), and investigated for use in the algal biophotovoltaic (BPV) device. The highest maximum power density was registered in the rGO-based BPV device (0.538 ± 0.014 mW m−2). This illustrates the potential of this local algal strain for use in BPV devices to generate bioelectricity in both the light and dark conditions.

To test the potential efficacy of recombinant macrophage inhibitory cytokine-1 (MIC-1/GDF15) as an obesity therapeutic. Male C57BL/6 J mice, either fed on normal chow or high-fat diet for 16 weeks to induce diet-induced obesity, were infused with either recombinant MIC-1/GDF15 or vehicle for 34 days by osmotic minipump. During the experimental period metabolic parameters were measured. Blood and tissue were collected for analysis of inflammatory markers. MIC-1/GDF15 decreased food intake and body weight of high-fat-fed and chow-fed mice compared with their vehicle-treated control mice. MIC-1/GDF15 reduced body weight, accompanied by greater reduction in fat mass in high-fat-fed mice compared to its effect on chow-fed mice. Further, whilst MIC-1/GDF15-treated chow-fed mice lost lean as well as fat mass, MIC-1/GDF15-treated high-fat-fed mice lost fat mass alone. This reduction in body weight and adiposity was due largely to reduced food intake, but MIC-1/GDF15-treated high-fat-fed mice also displayed increased energy expenditure that may be due to increased thermogenesis. MIC-1/GDF15-treated high-fat-fed mice also had higher circulating level of adiponectin and lower tissue expression, and circulating levels of leptin and inflammatory mediators associated with insulin resistance. Peripheral insulin and glucose intolerance were improved in both MIC-1/GDF15-treated high-fat-fed and chow-fed mice compared to that of their vehicle-treated control mice. MIC-1/GDF15 is highly effective in reducing adiposity and correcting the metabolic dysfunction of mice with high-fat fed. These studies suggest that MIC-1/GDF15 may be a candidate anti-obesity therapeutic.

We report for the first time a photosynthetically active algae immobilized in alginate gel within a fuel cell design for generation of bioelectricity. The algal-alginate biofilm was utilized within a biophotovoltaics (BPV) device developed for direct bioelectricity generation from photosynthesis. A peak power output of 0.289 mWm−2 with an increase of 18% in power output compared to conventional suspension culture BPV device was observed. The increase in maximum power density was correlated to the maximum relative electron transport rate (rETRm). The semi-dry type of photosynthetically active biofilm proposed in this work may offer significantly improved performances in terms of fuel cell design, bioelectricity generation, oxygen production and CO2 reduction.

The surface optimization and structural characteristics of Langmuir–Blodgett (LB) reduced graphene oxide thin (rGO) film treated by argon plasma treatment were studied. In this work, six times deposition of rGO was deposited on a clean glass substrate using the LB method. Plasma technique involving a variation of plasma power, i.e., 20, 60, 100 and 140 W was exposed to the LB-rGO thin films under argon ambience. The plasma treatment generally improves the wettability or hydrophilicity of the film surface compared to without treatment. Maximum wettability was observed at a plasma power of 20 W, while also increasing the adhesion of the rGO film with the glass substrate. The multilayer films fabricated were characterized by means of spectroscopic, structural and electrical studies. The treatment of rGO with argon plasma was found to have improved its biocompatibility, and thus its performance as an electrode for biophotovoltaic devices has been shown to be enhanced considerably.

Infection with dengue virus (DV) can result in dengue hemorrhagic fever and dengue shock syndrome, where patients suffer from bleeding and plasma leakage involving endothelial cells. Angiopoietins (Ang) 1 and 2 are important angiogenic factors that affect endothelial barrier integrity. In this study, DV was observed to induce endothelial leakage at multiplicity of infection of 10 in primary human umbilical vein endothelial cells (HUVEC) with interendothelial gap formation. Immunostaining of vascular endothelial cadherin (VE-cadherin) and zona occludin 1 (ZO-1) showed the absence of these endothelial junctional proteins at the cell-cell contact zones between adjacent cells. In addition, Ang1 that is required for protecting against endothelial hyperpermeability was found to be down-regulated during DV infection. Treatment with increasing concentrations of recombinant Ang1 was shown to prevent DV-induced endothelial hyperpermeability in a dose-dependent manner by preventing the down-regulation of VE-cadherin and ZO-1 at cell membrane. In contrast, the expression of Ang2, the natural antagonist of Ang1, was observed to be up-regulated during DV infection. Recombinant Ang2 added to HUVEC at non-toxic concentrations showed decreased in transendothelial electrical resistance reading and the down-regulation of VE-cadherin and ZO-1. These findings suggest that DV reduces the expression of Ang1 and enhances the expression of Ang2 in endothelial cells and that this imbalance of Ang 1 and Ang 2 may play a contributing role to the increased permeability of human primary endothelial cells during DV infection.

Altered bile flow physiology leads to many complications commonly seen in patients with cholestatic liver disease, regardless of the etiology. For each individual patient, a coordinated and effective treatment strategy must address the presence and the severity spectrum of malabsorption, malnutrition, vitamin and micronutrient deficiencies, pruritus, xanthomata, ascites, and liver failure, which are attributed directly or indirectly to diminished bile flow. An aggressive approach to maximizing the nutritional status of the child is vital to ensure optimal growth and development. Protein-calorie and/or fat supplementation is best discussed early. Decreasing the percentage of dietary long-chain triglycerides, providing medium-chain triglycerides, and ensuring adequate essential fatty acid and adequate protein intake may be helpful. Fat-soluble vitamin (A, D, E, and K) levels and micronutrient levels must be carefully and serially monitored and supplemented as necessary. Because the mechanisms that mediate pruritus of cholestasis remain to be determined, the use of empirical therapies continues to be standard practice. Ursodeoxycholic acid may ameliorate pruritus. Antihistamines and rifampicin may also provide temporary relief for some children. Based on the evidence that increased central opioidergic tone is present in chronic cholestasis, the use of opiate antagonists is promising but has not been evaluated in children. Selected patients with refractory pruritus that have failed maximal medical therapy have benefited from partial external biliary diversion. Ongoing dialogue with the family regarding the indications for liver transplantation is reasonable. Optimization and adherence with the pretransplant medical management enhance the chances for a successful outcome from liver transplantation. Specific to the pediatric patient, optimizing growth, development and nutrition, minimizing discomfort and disability, and aiding the child and family in coping with the stress, social, and emotional effects of chronic liver disease remain paramount.

The present paper reviews the applications and recent developments of microwave digestion or extraction as a sample preparation technique. This technique has been applied successfully in both organic and inorganic determinations for various sample matrices. It offers the benefits of rapid sample preparation, ease of automation and reduction in background contamination. Good recoveries are generally obtained.

BackgroundIn 2001 and 2002, fatal myocarditis resulted in the sudden deaths of four, two adult and two juvenile, orang utans out of a cohort of 26 in the Singapore Zoological Gardens.MethodsOf the four orang utans that underwent post-mortem examination, virus isolation was performed from the tissue homogenates of the heart and lung obtained from the two juvenile orang utans in Vero cell cultures. The tissue culture fluid was examined using electron microscopy. Reverse transcription and polymerase chain reaction with Encephalomyocarditis virus (EMCV)-specific primers targeting the gene regions of VP3/VP1 and 3D polymerase (3Dpol) confirmed the virus genus and species. The two EMCV isolates were sequenced and phylogenetic analyses of the virus genes performed. Serological testing on other animal species in the Singapore Zoological Gardens was also conducted.ResultsElectron microscopy of the two EMCV isolates, designated Sing-M100-02 and Sing-M105-02, revealed spherical viral particles of about 20 to 30 nm, consistent with the size and morphology of members belonging to the familyPicornaviridae. In addition, infected-Vero cells showed positive immunoflorescence staining with antiserum to EMCV. Sequencing of the viral genome showed that the two EMCV isolates were 99.9% identical at the nucleotide level, indicating a similar source of origin. When compared with existing EMCV sequences in the VP1 and 3Dpol gene regions, the nucleotide divergence were at a maximum of 38.8% and 23.6% respectively, while the amino acid divergence were at a maximum of 33.9% and 11.3% respectively. Phylogenetic analyses of VP1 and 3Dpol genes further grouped the Sing-M100-02 and Sing-M105-02 isolates to themselves, away from existing EMCV lineages. This strongly suggested that Sing-M100-02 and Sing-M105-02 isolates are highly divergent variants of EMCV. Apart from the two deceased orang utans, a serological survey conducted among other zoo animals showed that a number of other animal species had neutralizing antibodies to Sing-M105-02 isolate, indicating that the EMCV variant has a relatively wide host range.ConclusionsThe etiological agent responsible for the fatal myocarditis cases among two of the four orang utans in the Singapore Zoological Gardens was a highly divergent variant of EMCV. This is the first report of an EMCV infection in Singapore and South East Asia.

West Nile (Sarafend) virus was distinctly observed to bud from the plasma membrane rather than mature intracellularly. This has been observed with transmission electron microscopy. Using conventional scanning electron microscopy, budding at the plasma membrane especially at the filopodia was clearly illustrated. Immunogold labelling against the virus envelope protein was also performed to confirm this mode of exit. The gold particles were observed to be located at the sites where virus budding was seen under the field emission scanning electron microscope.

Background Dengue virus (DENV) is the causative agent of Dengue fever and the life-threatening Dengue Haemorrhagic fever or Dengue shock syndrome. In the absence of anti-viral agents or vaccine, there is an urgent need to develop an effective anti-viral strategy against this medically important viral pathogen. The initial interplay between DENV and the host cells may represent one of the potential anti-viral targeting sites. Currently the involvements of human membrane trafficking host genes or factors that mediate the infectious cellular entry of dengue virus are not well defined. Results In this study, we have used a targeted small interfering RNA (siRNA) library to identify and profile key cellular genes involved in processes of endocytosis, cytoskeletal dynamics and endosome trafficking that are important and essential for DENV infection. The infectious entry of DENV into Huh7 cells was shown to be potently inhibited by siRNAs targeting genes associated with clathrin-mediated endocytosis. The important role of clathrin-mediated endocytosis was confirmed by the expression of well-characterized dominant-negative mutants of genes in this pathway and by using the clathrin endocytosis inhibitor chlorpromazine. Furthermore, DENV infection was shown to be sensitive to the disruption of human genes in regulating the early to late endosomal trafficking as well as the endosomal acidic pH. The importance and involvement of both actin and microtubule dynamics in mediating the infectious entry of DENV was also revealed in this study. Conclusions Together, the findings from this study have provided a detail profiling of the human membrane trafficking cellular genes and the mechanistic insight into the interplay of these host genes with DENV to initiate an infection, hence broadening our understanding on the entry pathway of this medically important viral pathogen. These data may also provide a new potential avenue for development of anti-viral strategies and treatment of DENV infection.

Scanning electron microscopy of Chilomastix mesnili shows that the cysts are lemon-shaped with one end broadly rounded and the other conical. The trophozoite has five flagella coming out of the anterior end. Four of these are free and the fifth is attached to the body by an undulating membrane. The undulating membrane extends along the whole length of the body of the parasite with the exception of the tail. The tail is an elongated structure almost equal in length to the main body of the parasite.

The surface coat of Blastocystis hominis was studied in the electron microscope. In some cells the surface coat was seen in two layers; the external layer was more electron-dense and fragmented than the internal layer. It appears that the surface coat is being continuously formed by the parasite and shed in the environment. The fibrillar material of the surface coat attaches to the bacteria, in some cases, completely surrounding them, possibly causing cytoplasmic damage to the bacterial cell as indicated by loss of electron density.

The ultrastructure of the Iodamoeba bütschlii cyst from human feces was studied. The glycogen mass appears as a compact dense body in the cytoplasm without any surrounding membrane. The cytoplasm has no mitochondrion. The nucleus shows a distinct nucleolus filled with electron-dense particles. On one side of the nucleolus are electron-dense cytoplasmic masses measuring 200-400 nm. The nuclear membrane is two-layered and shows pores.

Microscopic imaging of viruses and their interactions with and effects on host cells are frequently held back by limitations of the microscope's resolution or the invasive nature of the sample preparation procedures. It is also difficult to have a technique that would allow simultaneous imaging of both surface and sub-surface on the same cell. This has hampered endeavours to elucidate virus-host interactions. Atomic Force Microscopy (AFM), which is commonly used in the physical sciences, is now becoming a good correlative form of microscopy used to complement existing optical, confocal and electron microscopy for biological applications In this study, the West Nile (Sarafend) virus-infected Vero cell model was used. The atomic force microscope was found to be useful in producing high resolution images of virus-host events with minimal sample processing requirements. The AFM was able to image the budding of the West Nile (Sarafend) virus at the infected cell surface. Proliferation of the filopodia and thickening of clusters of actin filaments accompanied West Nile virus replication. The study shows that the AFM is useful for virus-host interaction studies. The technique provides morphological information on both the virus and the host cell during the infection stages.

The sites of replication and of accumulation of viral macromolecules were examined using fluorescent antibodies to viral products and to cell organelles. Synthesis of envelope protein and its accumulation in a narrow rim around the nucleus were detected at 4 hours post infection; concurrently, a progressive change was observed in the rough endoplasmic reticulum from a fine to a coarse network emanating from around the nucleus. This change in the network was visible by light microscopy. The immunofluorescent sites of viral RNA synthesis, located by use of anti-double stranded RNA, extended from the perinuclear region in another fine network which included many small foci or vesicles; these sites were also visible by light microscopy late in infection. None of these changes were associated with any visible redistribution of actin, intermediate filaments or microtubules, and no nuclear involvement was detected. However, when microtubules were disrupted by vinblastine treatment of cells, the distribution of the immunofluorescent sites of viral RNA synthesis was modified and the virus yield was reduced by at least 10-fold. These results confirmed our biochemical studies showing separation of viral sites of RNA synthesis and translation, and the accumulation of envelope protein in nuclear-associated membranes. The relevance of these observations is discussed in relation to the reports of specific membrane structures induced in flavivirus-infected cells.

High-Level Expression and In Vivo Processing of Chimeric Ubiquitin Fusion Proteins in Saccharomyces Cerevisiae

Induction of microtubulin paracrystals (Pc) by Kunjin (KUN) virus occurred after 15 hours post-infection and were often associated with convoluted membranes (cm) and virus particles. Vinblastine sulphate which disrupts microtubulin, had an inhibitory effect on the virus production when added during the viral latent period. When infected samples were extracted with Triton X-100 and analysed by SDS-PAGE, four viral proteins were observed.

The results obtained from electron microscopy, adsorbed and internalised virus assays and immunofluorescence studies supported that the most likely mode of entry of Kunjin virus into Vero cells was by receptor-mediated endocytosis. This was deduced indirectly from the time sequence of events that occurred. Electron microscopy revealed that endocytosis of the virus through coated vesicles had occurred. The adsorbed and internalised virus assay and immunofluorescence studies showed that there were two factors being recycled during endocytosis: the receptor for the virus and clathrin, the protein found on coated pits and vesicles. The study showed that clathrin was recycled first, followed by the receptor.

The reverse transcriptase (RT) domain of the human immunodeficiency virus (HIV) pol gene was expressed in yeast. Purification and analysis of the recombinant protein indicated that yeast post–translational processing of RT was similar, if not identical, to that which had previously been observed in HIV infected lymphoid cells. In addition, recombinant RT was enzymatically active and could be inhibited by several anti–retroviral agents. These studies indicate that HIV RT expressed in yeast will be useful for the in vitro analysis of further potential inhibitors of this important enzyme.