Associations of <Emphasis Type="Italic">CTLA4</Emphasis> +49 A/G Dimorphism and HLA-DRB1*/DQB1* Alleles With Type 1 Diabetes from South India

The aim of present study was to elucidate the association of CTLA4 +49 A/G and HLA-DRB1*/DQB1* gene polymorphism in south Indian T1DM patients. The patients and controls (n?=?196 each) were enrolled for CTLA4 and HLA-DRB1*/DQB1* genotyping by RFLP/PCR-SSP methods. The increased frequencies of CTLA4 ‘AG’ (OR?=?1.99; p?=?0.001), ‘GG’ (OR?=?3.94; p?=?0.001) genotypes, and ‘G’ allele (OR?=?2.42; p?=?9.26?×?10^?8) were observed in patients. Reduced frequencies of ‘AA’ (OR?=?0.35; p?=?7.19?×?10^?7) and ‘A’ (OR?=?0.41; p?=?9.26?×?10^?8) in patients revealed protective association. Among HLA-DRB1*/DQB1* alleles, DRB1*04 (OR?=?3.29; p?=?1.0?×?10^?5), DRB1*03 (OR?=?2.81; p?=?1.9?×?10^?6), DQB1*02:01 (OR?=?2.93; p?=?1.65?×?10^?5), DQB1*02:02 (OR?=?3.38; p?=?0.0003), and DQB1*03:02 (OR?=?7.72; p?=?0.0003) were in susceptible association. Decreased frequencies of alleles, DRB1*15 (OR?=?0.32; p?=?2.55?×?10^?7), DRB1*10 (OR?=?0.45; p?=?0.002), DQB1*06:01 (OR?=?0.43; p?=?0.0001), and DQB1*05:02 (OR?=?0.28; p?=?2.1?×?10^?4) in patients were suggested protective association. The combination of DRB1*03+AG (OR?=?5.21; p?=?1.4?×?10^?6), DRB1*04+AG (OR?=?2.14; p?=?0.053), DRB1*04+GG (OR?=?5.21; p?=?0.036), DQB1*02:01+AG (OR?=?4.44; p?=?3.6?×?10^?5), DQB1*02:02+AG (OR?=?20.9; p?=?9.5?×?10^?4), and DQB1*02:02+GG (OR?=?4.06; p?=?0.036) revealed susceptible association. However, the combination of DRB1*10+AA (OR?=?0.35; p?=?0.003), DRB1*15+AA (OR?=?0.22; p?=?5.3?×?10^?7), DQB1*05:01+AA (OR?=?0.45; p?=?0.007), DQB1*05:02+AA (OR?=?0.17; p?=?1.7?×?10^?4), DQB1*06:01+AA (OR?=?0.40; p?=?0.002), and DQB1*06:02+AG (OR?=?0.34; p?=?0.001) showed decreased frequency in patients, suggesting protective association. In conclusion, CTLA4/HLA-DR/DQ genotypic combinations revealed strong susceptible/protective association toward T1DM in south India. A female preponderance in disease associations was also documented.     

Growth and phosphorus removal by <Emphasis Type="Italic">Synechococcus elongatus</Emphasis> co-immobilized in alginate beads with <Emphasis Type="Italic">Azospirillum brasilense</Emphasis>

This study examined the co-immobilization of the cyanobacterium Synechococcus elongatus with the plant growth-promoting bacterium Azospirillum brasilense in alginate beads and its potential application for the removal of phosphorus from aquaculture wastewater. Co-immobilization of both microorganisms significantly increased the cell density of S. elongatus (2852.5?×?10⁴ cells mL^?1) compared with that of immobilization of cyanobacteria alone (1325.2?×?10⁴ cells mL^?1). Chlorophyll a content was similar in co-immobilized (11.1?±?3.5 pg cell^?1) and immobilized S. elongatus (14.5?±?4.9 pg cell^?1). Azospirillum brasilense showed continuous growth until day 2, after which its cell concentration declined until the end of the assay. Co-immobilized S. elongatus removed more phosphorus (44.8 %) than immobilized cyanobacteria cells alone (32.0 %). In conclusion, phosphate removal was greater with free cells of S. elongatus but overlapped with the values that were obtained with the treatment of co-immobilization of cells. Our results demonstrate that A. brasilense enhances the growth of S. elongatus and improves its removal of phosphorus when they are co-immobilized in alginate beads compared with only immobilization of cyanobacteria cells alone.     

Genetic risk factors for myocardial infarction more clearly manifest for early age of first onset

Epidemiological genetics established that heritability in determining the risk of myocardial infarction (MI) is substantially greater when MI occurs early in life. However, the genetic architecture of early-onset and late-onset MI was not compared. We analyzed genotype frequencies of SNPs in/near 20 genes whose protein products are involved in the pathogenesis of atherosclerosis in two groups of Russian patients with MI: the first group included patients with age of first MI onset <60 years (N?=?230) and the second group with onset ≥60 years (N?=?174). The control group of corresponding ethnicity consisted of 193 unrelated volunteers without cardiovascular diseases (93 individuals were over 60 years). We found that in the group of patients with age of onset <60 years, SNPs FGB rs1800788*T, TGFB1 rs1982073*T/T, ENOS rs2070744*C and CRP rs1130864*T/T were associated with risk of MI, whereas in patients with age of onset ≥60 years, only TGFB1 rs1982073*T/T was associated with risk of MI. Using APSampler software, we found composite markers associated with MI only in patients with early onset: FGB rs1800788*T?+?TGFB1 rs1982073*T; FGB rs1800788*T?+?LPL rs328*C?+?IL4 rs2243250*C; FGB rs1800788*T?+?ENOS rs2070744*C (Fisher p values of 1.4?×?10^?6 to 2.2?×?10^?5; the permutation p values of 1.1?×?10^?5 to 3.0?×?10^?4; ORs?=?2.67–2.54). Alleles included in the combinations were associated with MI less significantly and with lower ORs than the combinations themselves. The result showed a substantially greater contribution of the genetic component in the development of MI if it occurs early in life, and demonstrated the usefulness of genetic testing for young people.     

Multi-spectroscopic studies on the interaction between traditional Chinese herb,helicid with pepsin

Study on the binding properties of helicid by pepsin systematically using multi-spectroscopic techniques and molecular docking method, and these interactions comprise biological recognition at molecular level and backbone of biological significance in medicine concerned with the uses, effects, and modes of action of drugs. We investigated the mechanism of interaction between helicid and pepsin by using various spectroscopic techniques viz., fluorescence spectra, UV–Vis absorption spectra, circular dichroism (CD), 3D spectra, synchronous fluorescence spectra and molecular docking methods. The quenching mechanism associated with the helicid–pepsin interaction was determined by performing fluorescence measurements at different temperatures. From the experimental results show that helicid quenched the fluorescence intensity of pepsin via a combination of static and dynamic quenching process. The binding constants (K_a) at three temperatures (288, 298, and 308 K) were 7.940?×?10⁷, 2.082?×?10⁵ and 3.199?×?10⁵ L mol^?1, respectively, and the number of binding sites (n) were 1.44, 1.14, and 1.18, respectively. The n value is close to unity, which means that there is only one independent class of binding site on pepsin for helicid. Thermodynamic parameters at 298 K were calculated as follows: ΔH^o (??83.85 kJ mol^?1), ΔG^o (??33.279 kJ mol^?1), and ΔS^o (??169.72 J K^?1 mol^?1). Based on thermodynamic analysis, the interaction of helicid with pepsin is driven by enthalpy, and Van der Waals’ forces and hydrogen bonds are the main forces between helicid and pepsin. A molecular docking study further confirmed the binding mode obtained by the experimental studies. The conformational changes in the structure of pepsin was confirmed by 3D fluorescence spectra and circular dichroism.     

Genome size and ploidy level among wild and cultivated <Emphasis Type="Italic">Prunus</Emphasis> taxa in Slovakia

2C DNA content and ploidy level variation of Prunus spinosa and closely related taxa together with Prunus domestica L. and Prunus insititia L. was studied in Slovakia. The aim of the study was to define genome sizes and find differences between closely related taxa within Prunus spinosa sensu lato mentioned in previous works. According to our results, investigated taxa can be divided into three groups according to ploidy level: Prunus spinosa, Prunus dasyphylla, Prunus ×fruticans, Prunus ×dominii and Prunus ×schurii are tetraploids, Prunus ×fechtneri is pentaploid, and P. domestica and P. insititia are hexaploids. Genome size differences within tetraploid taxa were relatively small (Prunus spinosa: 1.40?±?0.02, P. ×domini: 1.44?±?0.01, P. ×fruticans: 1.48?±?0.02, P. ×schurii: 1.44?±?0.02), but statistically significant. Although further research is needed, it seems that the concept of several taxa as product of hybridization between P. spinosa and cultivated plum species has been supported by our study.     

Inheritance pattern and genetic correlations among growth and wood quality traits in Para rubber tree (<Emphasis Type="Italic">Hevea brasiliensis</Emphasis>) and implications for breeding

Inheritance pattern of wood traits viz. specific gravity, fibre dimensions and fibre-derived biometrical indices and their interactions among themselves and with that of growth are reported in Hevea brasiliensis. Girth (h² =???0.02?±?0.44 to h² =?0.35?±?0.24) showed moderate genetic control. Among wood traits, specific gravity (h²?=?0.15?±?0.31 to h² =?0.33?±?0.28) was found to be under moderate genetic control. Fibre traits viz., fibre length (h² =???0.26?±?0.30 to h² =?0.50?±?0.34), fibre diameter (h² =?0.19?±?0.49 to h² =?0.70?±?0.11), fibre lumen diameter (h² =???0.18?±?0.35 to h² =?0.56?±?0.47) and fibre wall thickness (h² =???5.17?±?5.26 to h² =?0.50?±?0.50) were under moderate to strong genetic control. Among fibre-derived indices, flexibility coefficient (h² =?0.48?±?0.21 to h² =?0.89?±?0.29) showed moderate to very strong genetic control. The Runkel ratio (h² =???0.40?±?0.27 to h² =?0.42?±?0.29) and slenderness ratio (h² =???0.36?±?0.29 to h² =?0.43?±?0.28) showed moderate genetic control. Girth showed very strong positive genetic correlation with fibre wall thickness and strong positive correlation with fibre width indicating scope of indirect selection potential for these traits. Wood specific gravity was not correlated with either girth or fibre traits. Hence, it would be possible to concomitantly improve growth and fibre traits without adversely affecting wood specific gravity. Moderate to very high estimates of heritability for fibre traits, girth and specific gravity indicated that considerable genetic gain can be realised for these traits. Implications of the above findings in genetic improvement of wood in Hevea are discussed.     

Abscisic Acid-Induced Starch Accumulation in Bioenergy Crop Duckweed <Emphasis Type="Italic">Spirodela polyrrhiza</Emphasis>

Spirodela polyrrhiza, a fast-growing duckweed with high starch and low lignin content, shows promise as a feedstock for bioenergy. Abscisic acid (ABA) is a biological hormone that controls plant growth and stress response. The effects of different ABA concentrations (0, 1.0 × 10^?5, 1.0 × 10^?4, 1.0 × 10^?3, 1.0 × 10^?2, and 1.0 × 10^?1 mg/L) on duckweed biomass growth, carbon dioxide fixation, formation of photosynthetic pigments (Chlorophyll a (Chla), Chlorophyll b (Chlb), and carotenoids), the activities of soluble starch synthase (SSS) and starch branching enzyme (SBE), and the starch content of biomass were investigated in this study. ABA at concentrations lower than 1.0 × 10^?3 mg/L promoted carbon dioxide fixation, whereas it inhibited carbon dioxide fixation at concentrations over 1.0 × 10^?3 mg/L. ABA enhanced SSS and SBE activities at concentrations lower than 1.0 × 10^?2 mg/L. ABA treatment increased the content of Chla, Chlb, and carotenoids and resulted in the enhancement of starch content. Chla content gradually increased with the increasing concentration of ABA (1.0 × 10^?5 to 1.0 × 10^?2 mg/L). After culturing for 10 days, starch content in 1.0 × 10^?2 mg/L ABA medium reached 35.3% of dry weight (DW), which was the highest level in this study. This suggests that there is a great potential to develop a technology to increase starch accumulation in duckweed which can be used as an alternative to corn, sugarcane, or other food crops as a starch source.     

Boosting and Localizing Near-Field in Plasmonic Mirror-Image Nanoepsilon

A novel plasmonic mirror-image nanoepsilon (MINE) structure is studied to achieve significantly enhanced and localized near-field. It is well-known that the nanorod dimer is able to gain a high local field at the center of the structure by adjusting its rod width, rod length, and gap distance. When adopting an auxiliary nanoring structure electron reservoirs surrounding the nanorod-dimer to form the MINE structure, the local field can be further enhanced owing to a large amount of charges into sharp dimer structure which can confine the accumulation of charges to apexes. Thus, better synergistic interaction of gap effect and lightning-rod effect can be achieved for high field enhancement around nanoscale gap. The symmetric mode in the MINE structure enables strong near-field with a concentrated distribution around the gap. The influences of rod-tip angle and gap distance on the optical properties of plasmonic MINE structures are numerically investigated. With reducing the rod-tip angle and gap distance, considerable enhancements on the field intensities at the rod tips and the gap center can be attributed to the improved lightning-rod and gap effects. The near-field intensities at the rod tip and gap center are dramatically enlarged ~1.94 × 10⁴ and ~1.41 × 10⁴ times with the rod-tip angle of 41° and gap distance of 10 nm, and the near-field is localized within an extremely small range. These features are very beneficial for various plasmonic applications.     

Bayesian logistic regression in detection of gene–steroid interaction for cancer at <Emphasis Type="Italic">PDLIM5</Emphasis> locus

The PDZ and LIM domain 5 (PDLIM5) gene may play a role in cancer, bipolar disorder, major depression, alcohol dependence and schizophrenia; however, little is known about the interaction effect of steroid and PDLIM5 gene on cancer. This study examined 47 single-nucleotide polymorphisms (SNPs) within the PDLIM5 gene in the Marshfield sample with 716 cancer patients (any diagnosed cancer, excluding minor skin cancer) and 2848 noncancer controls. Multiple logistic regression model in PLINK software was used to examine the association of each SNP with cancer. Bayesian logistic regression in PROC GENMOD in SAS statistical software, ver. 9.4 was used to detect gene–steroid interactions influencing cancer. Single marker analysis using PLINK identified 12 SNPs associated with cancer (P < 0.05); especially, SNP rs6532496 revealed the strongest association with cancer (P = 6.84 × 10^?3); while the next best signal was rs951613 (P = 7.46 × 10^?3). Classic logistic regression in PROC GENMOD showed that both rs6532496 and rs951613 revealed strong gene–steroid interaction effects (OR = 2.18, 95% CI = 1.31?3.63 with P = 2.9 × 10^?3 for rs6532496 and OR = 2.07, 95% CI = 1.24 ?3.45 with P = 5.43 × 10^?3 for rs951613, respectively). Results from Bayesian logistic regression showed stronger interaction effects (OR = 2.26, 95% CI = 1.2 ?3.38 for rs6532496 and OR = 2.14, 95% CI = 1.14 ?3.2 for rs951613, respectively). All the 12 SNPs associated with cancer revealed significant gene–steroid interaction effects (P < 0.05); whereas 13 SNPs showed gene–steroid interaction effects without main effect on cancer. SNP rs4634230 revealed the strongest gene–steroid interaction effect (OR = 2.49, 95% CI = 1.5 ?4.13 with P = 4.0 × 10^?4 based on the classic logistic regression and OR = 2.59, 95% CI = 1.4 ?3.97 from Bayesian logistic regression; respectively). This study provides evidence of common genetic variants within the PDLIM5 gene and interactions between PLDIM5 gene polymorphisms and steroid use influencing cancer.     

Changes in Immunity,Expression of some Immune-Related Genes of Shabot Fish, <Emphasis Type="Italic">Tor grypus</Emphasis>, Following Experimental Infection with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>: Effects of Autochthonous Probiotics

In this study, the effects of orally administrated two native probiotics (Lactobacillus plantarum and Lactobacillus delbrueckii ssp. bulguricus), isolated from the intestine of Shabot fish, Tor grypus, on some immune response parameters and immune-related genes expression against Aeromonas hydrophila in T. grypus were evaluated. Four hundred and eighty juveniles weighing 45?±?10 g were randomly divided into four groups (with three replications) and fed with the experimental diet containing 5?×?10⁷ cfu g^?1 of L. plantarum (G1), Lactobacillus bulgaricus (G2), Lactobacillus casei (G3), and a control diet (without probiotics) for 60 continuous days. At the end of the dietary treatments, fish were challenged with a lethal concentration of A. hydrophila (5?×?10⁸ CFU ml^?1) via intra peritoneal (i.p) injection. Blood and head kidney samples were taken from six fish in each treatment before challenging and 6, 12, 24, and 48 h and also 7 days after injection. The results showed that lysozyme, complement, bactericidal, and NBT activity of probiotic-treated groups were significantly elevated (P?<?0.05). The IL-8, IL-1β, and TNF-α gene expressions were significantly higher in all probiotic-treated groups (P?<?0.05). Meanwhile, a high direct correlation was observed between serum immune parameters and expression of immune-related genes (P?<?0.0001); furthermore, the highest correlation (R ²?=?0.634, P?<?0.0001) was recorded between IL-1β expression and NBT activity. It can be concluded that not only two native probiotics strains stimulate serum immune responses parameters and immune-related gene expression in T. grypus, but also a high correlation was seen among these indices. The study suggests that gastrointestinal colonization is preferred for host specificity as the strain previously derived from shabot fish displayed better colonization than the non-indigenous bacteria strain such as L. casei. Therefore, these native probiotics bacteria can be accounted as suitable candidates to immune stimulation in fish.     

Presence of pathogenic bacteria in ice cubes and evaluation of their survival in different systems

In this study, 60 samples of ice cubes produced at different levels (domestic, restaurant and industrial facilities), within a restricted geographical area, were investigated for their general microbiological characteristics through the analysis of populations other than enteric bacteria. Total mesophilic bacteria were in the range 1.01?×?10²–9.55?×?10³, 3.12?×?10²–6.31?×?10³ and 1.30?×?10²–3.99?×?10³?CFU/100 mL of thawed ice from domestic freezer (DF), stock boxes (SB) for self-production performed with ice machines in bars and pubs, and from sales packages (SP) of industrial productions, respectively. Some DF and SP samples were negative for the presence of total psychrotrophic bacteria, showing that there are no specific microbial groups associated with ice. Pseudomonads were found in the majority of ice samples analyzed. The levels of contamination of the ice samples were significantly different among the three ice cube production levels. The samples produced at domestic level and those collected from bars and pubs were characterised by the highest cell densities. The colonies representative for the different bacterial morphologies were randomly picked up from plates, purified to homogeneity and subjected to the phenotypic and genotypic characterisation. Fifty-two strains representing 31 species of eight bacterial genera were identified, with the most numerous groups included in Pseudomonas, Staphylococcus, Bacillus and Acinetobacter. A consistent percentage of the microorganisms identified from ice are known agents of human infections, and their presence indicate an environmental contamination. In order to evaluate the effectiveness of the ice cubes to transfer pathogenic agents to consumers, a bar consumption was simulated with different drink systems added with ice cubes artificially contaminated with the strains found at dominant levels (Acinetobacter lwoffii ICE100, Bacillus cereus ICE170, Pseudomonas putida ICE224 and Staphylococcus haemolyticus ICE182), and the results showed a consistent reduction of bacterial risk due to alcohol, CO₂, pH and antibacterial ingredients of vodka, whisky, Martini, peach tea, tonic water and coke.     

The nitrite reductase activity of horse heart carboxymethylated-cytochrome <Emphasis Type="Italic">c</Emphasis> is modulated by cardiolipin

Horse heart carboxymethylated cytc (CM-cytc) displays myoglobin-like properties. Here, the effect of cardiolipin (CL) liposomes on the nitrite reductase activity of ferrous CM-cytc [CM-cytc-Fe(II)], in the presence of sodium dithionite, is reported between pH 5.5 and 7.6, at 20.0 °C. Cytc-Fe(II) displays a very low value of the apparent second-order rate constant for the NO₂ ^?-mediated conversion of cytc-Fe(II) to cytc-Fe(II)-NO [k _on = (7.3 ± 0.7) × 10^?2 M^?1 s^?1; at pH 7.4], whereas the value of k _on for NO₂ ^? reduction by CM-cytc-Fe(II) is 1.1 ± 0.2 M^?1 s^?1 (at pH 7.4). CL facilitates the NO₂ ^?-mediated nitrosylation of CM-cytc-Fe(II) in a dose-dependent manner, the value of k _on for the NO₂ ^?-mediated conversion of CL–CM-cytc-Fe(II) to CL–CM-cytc-Fe(II)-NO (5.6 ± 0.6 M^?1 s^?1; at pH 7.4) being slightly higher than that for the NO₂ ^?-mediated conversion of CL–cytc-Fe(II) to CL–cytc-Fe(II)-NO (2.6 ± 0.3 M^?1 s^?1; at pH 7.4). The apparent affinity of CL for CM-cytc-Fe(II) is essentially pH independent, the average value of B being (1.3 ± 0.3) × 10^?6 M. In the absence and presence of CL liposomes, the nitrite reductase activity of CM-cytc-Fe(II) increases linearly on lowering pH and the values of the slope of the linear fittings of Log k _on versus pH are ?1.05 ± 0.07 and ?1.03 ± 0.03, respectively, reflecting the involvement of one proton for the formation of the transient ferric form, NO, and OH^?. These results indicate that Met80 carboxymethylation and CL binding cooperate in the stabilization of the highly reactive heme-Fe atom of CL–CM-cytc.     

Evaluation of growth and phycobiliprotein composition of cyanobacteria isolates cultivated in different nitrogen sources

Phycobiliproteins, light-harvesting pigments found in cyanobacteria and in some eukaryotic algae, have numerous commercial applications in food, cosmetic, and pharmaceutical industries. Colorant production from cyanobacteria offers advantages over their production from higher plants, as cyanobacteria have fast growth rate and high photosynthetic efficiency and require less space. In this study, three cyanobacteria strains were studied for phycobiliprotein production and the influence of sodium nitrate, potassium nitrate and ammonium chloride on the growth and phycobiliprotein composition of the strains were evaluated. In the batch culture period of 12 days, Phormidium sp. and Pseudoscillatoria sp. were able to utilize all tested nitrogen sources; however, ammonium chloride was the best nitrogen source for both strains to achieve maximum growth rate μ?=?0.284?±?0.03 and μ?=?0.274?±?0.13 day^?1, chlorophyll a 16.2?±? 0.5 and 12.2?±? 0.2 mg L^?1, and phycobiliprotein contents 19.38?±?0.09 and 19.99?±?0.14% of dry weight, whereas, for Arthrospira platensis, the highest growth rate of μ?=?0.304?±?0.0 day^?1, chlorophyll a 19.1?±?0.5 mg L^?1, and phycobiliprotein content of 22.27?±?0.21% of dry weight were achieved with sodium nitrate. The phycocyanin from the lyophilized cyanobacterial biomass was extracted using calcium chloride and food grade purity (A₆₂₀/A₂₈₀ ratio >?0.7) was achieved. Furthermore, phycocyanin was purified using two-step chromatographic method and the analytical grade purity (A₆₂₀/A₂₈₀ ratio >?4) was attained. SDS-PAGE demonstrated the purity and presence of two bands corresponding to α- and β-subunits of the C-phycocyanin. The results showed that Phormidium sp. and Pseudoscillatoria sp. could be good candidates for phycocyanin production.     

Comparative analysis of prokaryotic diversity in solar salterns in eastern Anatolia (Turkey)

The prokaryotic communities of four salterns (Bingöl, Fadlum, Kemah, and Tuzlagözü) in Turkey were examined and compared using the cultivation and cultivation-independent methods [fluorescence in situ hybridization (FISH) and 454 pyrosequencing]. FISH analysis with universal probes revealed that feeding waters carried 1.6 × 10²–1.7 × 10³ cells mL^?1, while crystallization ponds carried 3.8 × 10⁶–2.0 × 10⁷ cells mL^?1 that were mostly haloarchaea, including square cells (except for Kemah). High-throughput 16S rRNA-based gene sequencing showed that the most frequent archaeal OTUs in Bingöl, Fadlum, Tuzlagözü, and Kemah samples were affiliated with Haloquadratum (76.8 %), Haloarcula (27.8 %), Halorubrum (49.6 %), and Halonotius (59.8 %), respectively. Bacteroidetes was the dominant bacterial phylum in Bingöl and Fadlum, representing 71.5 and 79.5 % of the bacterial OTUs (respectively), while the most abundant bacterial phylum found in the Kemah saltern was Proteobacteria (79.6 %). The majority of the bacterial OTUs recovered from Tuzlagözü belonged to the Cyanobacteria (35.7 %), Bacteroidetes (35.0 %), and Proteobacteria (25.5 %) phyla. Cultivation studies revealed that the archaeal isolates were closely related to the genera Halobacterium, Haloarcula, and Halorubrum. Bacterial isolates were confined to two phyla, Proteobacteria (Alphaproteobacteria and Gammaproteobacteria classes) and Bacteroidetes. Comparative analysis showed that members of the Euryarchaeota, Bacteroidetes, Proteobacteria, and Cyanobacteria phyla were major inhabitants of the solar salterns.     

A new biopesticide from a local <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> var. <Emphasis Type="Italic">tenebrionis</Emphasis> (Xd3) against alder leaf beetle (Coleoptera: Chrysomelidae)

Use of chemical pesticides in agriculture harms humans, non-target organisms and environments, and causes increase resistance against chemicals. In order to develop an effective bio-pesticide against coleopterans, particularly against Agelastica alni (Coleoptera: Chrysomelidae) which is one of the serious pests of alder leaf and hazelnut, we tested the insecticidal effect of 21 Bacillus isolates against the larvae and adults of the pest. Bacillus thuringiensis var. tenebrionis-Xd3 (Btt-Xd3) showed the highest insecticidal effect based on screening tests. For toxin protein production and high sporulation of Xd3, the most suitable medium, pH and temperature conditions were determined as nutrient broth medium enriched with salts, pH 7 and 30?°C, respectively. Sporulated Btt-Xd3 in nutrient broth medium enriched with salts transferred to fermentation medium containing soybean flour, glucose and salts. After fermentation, the mixture was dried in a spray dryer, and spore count of the powder product was determined as 1.6?×?10¹⁰ c.f.u. g^?1. Moisture content, suspensibility and wettability of the formulation were determined as 8.3, 86% and 21 s, respectively. Lethal concentrations (LC₅₀) of formulated Btt-Xd3 were determined as 0.15?×?10⁵ c.f.u. ml^?1 for larvae at laboratory conditions. LC₅₀ values were also determined as 0.45?×?10⁶ c.f.u. ml^?1 at the field condition on larval stage. Our results showed that a new bio-pesticide developed from B. thuringiensis tenebrionis (Xd3) (Btt-Xd3) may be valuable as a biological control agent for coleopteran pests.     

Studies on the interaction between benzidine and bovine serum albumin by spectroscopic methods

The interaction between bovine serum albumin (BSA) and benzidine (BD) in aqueous solution was investigated by fluorescence spectroscopy, circular dichroism (CD) spectra and UV–Vis spectroscopy, as well as resonance light scattering spectroscopy (RLS). It was proved from fluorescence spectra that the fluorescence quenching of BSA by BD was a result of the formation of BD–BSA complex, and the binding constants (K _a) were determined according to the modified Stern–Volmer equation. The enthalpy change (ΔH) and entropy change (ΔS) were calculated to be ?34.11 kJ mol^?1 and ?25.89 J mol^?1 K^?1, respectively, which implied that van der Waals force and hydrogen bond played predominant roles in the binding process. The addition of increasing BD to BSA solution caused the gradual enhancement in RLS intensity, exhibiting the forming of the aggregate. Moreover, the competitive experiments of site markers suggested that the binding site of BD to BSA was located in the region of subdomain IIA (sudlow site I). The distance (r) between the donor (BSA) and the acceptor (BD) was 4.44 nm based on the Förster theory of non–radioactive energy transfer. The results of synchronous fluorescence and CD spectra demonstrated the microenvironment and the secondary conformation of BSA were changed.     

Three new species of <Emphasis Type="Italic">Spadicoides</Emphasis> from Lushan Mountain,China

Spadicoides lushanensis sp. nov., S. multiseptata sp. nov. and S. rostrata sp. nov. are described and illustrated from specimens collected on dead branches of unidentified plants in Lushan Mountain, China. Spadicoides lushanensis is characterized by obclavate, rostrate, pale brown, 40–65?×?4.5–5.5 μm, smooth, 6–8-euseptate conidia. Spadicoides multiseptata is easily distinguished by obclavate, smooth, 11–15-euseptate, 65–115?×?11.5–13 μm, brown to pale brown conidia with a subacute apex. Spadicoides rostrata differs from other described Spadicoides species by obclavate, rostrate, pale brown, 40–65?×?10.5–12.5 μm, smooth, predominantly 5-euseptate conidia. A dichotomous key to Spadicoides species is provided.     

A novel alphabaculovirus isolated from the cotton bollworm, <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> (Hubner) (Lepidoptera: Noctuidae): characterization and pathogenicity

The cotton bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) is a polyphagous pest that exist all over the world. It is also very destructive in Turkey on various agricultural products. In this study, we have detected an alphabaculovirus from Helicoverpa armigera larvae collected from cotton field. The virus isolate was named as Helicoverpa armigera nucleopolyhedrovirus (HearNPV-O1) based on morphological and molecular properties. This is the first record of baculovirus from H. armigera in Eurasia region. Scanning electron microscopy examinations of polyhedral inclusion bodies (PIBs) showed their irregular morphology with average diameter of 0.85 to 1.25 μm. Transmission electron microscopy studies revealed that the nucleocapsids were multiple enveloped and bacilliform shaped. The size of nucleocapsid was found as 279 nm with a width of 56 nm. Digestion of viral genome by PstI generated 8 fragments with total size of 130.9 kbp. According to the phylogenetic analysis of the partial polyhedrin (polh), late expression factor 8 (lef8) and late expression factor 9 (lef9) genes, HearNPV-O1 isolate is very close to H. armigera MNPV Indian isolate-443. Five different concentrations of HearNPV-O1 between 10³ and 10⁷ were used in dose-response test on neonate, 3rd and 5th instars larvae of H. armigera. The highest dose (10⁷) showed 92%, 88% and 57% mortality, respectively within 14 days. LC₅₀ values of HearNPV-O1 isolate were calculated as 6?×?10⁴, 7?×?10⁴ and 8?×?10⁶ PIBs/mL^?1 against neonate, 3rd and 5th instars larval stages, respectively. These results demonstrate that HearNPV-O1 isolate may be a potential biocontrol agent to be utilized against H. armigera.     

Abundance and Diversity of Biofilms in Natural and Artificial Aquifers of the Äspö Hard Rock Laboratory,Sweden

Six cores were drilled and retrieved from 186-m depth in the Äspö Hard Rock Laboratory (HRL) tunnel to investigate whether indigenous biofilms develop on fracture surfaces in groundwater-conducting aquifers in granitic rock. A clone library was constructed from fracture surface material (FSM), for community composition analysis. Quantitative polymerase chain reaction (qPCR) was applied to quantify gene copies using the 16S rRNA gene for domain Bacteria and the adenosine-phosphosulfate reductase gene (apsA) for sulfate-reducing bacteria (SRB). Results were compared with three groundwater systems with biofilms in laminar flow reactors (LFRs) at 450-m depth in the Äspö HRL. The total number of cells, counted microscopically, was approximately 2?×?10⁵ cells cm^–2 in the LFR systems, consistent with the obtained qPCR 16S rRNA gene copies. qPCR analysis reported ～1?×?10² up to ～1?×?10⁴ gene copies cm^–2 on the FSM from the drill cores. In the FSM biofilms, 33% of the sequenced clones were related to the iron-reducing bacterium Stenotrophomonas maltophilia, while in the LFR biofilms, 41% of the sequenced clones were affiliated with the genera Desulfovibrio, Desulforhopalus, Desulfomicrobium, and Desulfobulbus. The community composition of the FSM biofilms differed from the drill water community, excluding drill water contamination. This work reports significant numbers of microorganisms on natural hard rock aquifer fracture surfaces with site-specific community compositions. The probability that biofilms are generally present in groundwater-conducting aquifers in deep granitic rock is consequently great.     

Cyanobacteria consumption by cladocerans: a case study on facilitation

Eutrophic systems are stressful for zooplankton species, especially for small organisms (<?1000 μm) that are inefficient in consuming large colonies or filaments of cyanobacteria. Certain mechanisms, however, enable organisms to coexist in spite of the stress related to poor food quality or manageability of the diet. The present work suggests that coprophagy is recurrent behavior in cladocerans that may facilitate the survival and growth of some species in eutrophic systems. We chose three clones of Moina macrocopa that inhabit eutrophic systems. The species selected as possible facilitators were the cladocerans Simocephalus vetulus and Daphnia similis, and the ostracod Heterocypris incongruens, because they are cosmopolitan, have high rates of cyanobacterial filtration and in some cases coexist with Moina. The design used for the demographic experiments of the three clones of M. macrocopa was based on the source of nutrition and consisted of a control diet of Scenedesmus acutus (1?×?10⁶ cells ml), a diet of undigested cyanobacteria (10?×?10⁶ cells ml), and this same diet after digestion by D. similis, S. vetulus or H. incongruens. The excreta of the cladocerans was a deficient diet for Moina, whose populations decreased by more than 1 individual per day. On the contrary, fecal pellets of the ostracod were a diet that allowed Moina to have positive population growth (0.3 day^?1). Our results help to explain the high densities achieved by cladocerans in eutrophic tropical water bodies where cyanobacterial blooms are normal.