Early Stages Of Culture Research Articles

The dynamics of bacterial community in a polyculture aquaculture system of Penaeus chinensis , Rhopilema esculenta and Sinonovacula constricta

Polyculture with Chinese shrimp Penaeus chinensis, jellyfish Rhopilema esculenta and razor clam Sinonovacula constricta (S.J.R) is a common aquaculture system in Dandong, China. Investigation on the composition and function of microbial communities and their relationship with environmental factors in aquaculture systems have crucial implications. Therefore, the sequencing of 16S rRNA gene was applied to profile the bacterial community composition of water and sediment in S.J.R polyculture system in the present study. Proteobacteria was the dominant phylum in S.J.R polyculture ponds following with Actinobacteria and Bacteroidetes in water; in contrast, more diverse phyla were detected in sediments. Water temperature, pH and dissolved oxygen were determined the bacterial community composition in the S.J.R polyculture systems. Specially, Planktomarina, PS1 clade, Candidatus Aquiluna and Fibrobacteres, which can resist the phytoplankton blooms or degrade the cellulose, were more abundant in water at the early stage of cultivation in the S.J.R polyculture system. Moreover, the functions of bacterial communities in water were mainly associated with cell communication mediated, while the functions related to basal metabolism were enriched in sediments. The findings of this study allowed us to better understand the bacterial community in the S.J.R polyculture system and accumulated data for the improvement of cultural management.

Tuning of Titanium Microfiber Scaffold with UV-Photofunctionalization for Enhanced Osteoblast Affinity and Function.

Titanium (Ti) is an osteoconductive material that is routinely used as a bulk implant to fix and restore bones and teeth. This study explored the effective use of Ti as a bone engineering scaffold. Challenges to overcome were: (1) difficult liquid/cell infiltration into Ti microfiber scaffolds due to the hydrophobic nature of Ti; and (2) difficult cell attachment on thin and curved Ti microfibers. A recent discovery of UV-photofunctionalization of Ti prompted us to examine its effect on Ti microfiber scaffolds. Scaffolds in disk form were made by weaving grade 4 pure Ti microfibers (125 µm diameter) and half of them were acid-etched to roughen the surface. Some of the scaffolds with original or acid-etched surfaces were further treated by UV light before cell culture. Ti microfiber scaffolds, regardless of the surface type, were hydrophobic and did not allow glycerol/water liquid to infiltrate, whereas, after UV treatment, the scaffolds became hydrophilic and immediately absorbed the liquid. Osteogenic cells from two different origins, derived from the femoral and mandibular bone marrow of rats, were cultured on the scaffolds. The number of cells attached to scaffolds during the early stage of culture within 24 h was 3–10 times greater when the scaffolds were treated with UV. The development of cytoplasmic projections and cytoskeletal, as well as the expression of focal adhesion protein, were exclusively observed on UV-treated scaffolds. Osteoblastic functional phenotypes, such as alkaline phosphatase activity and calcium mineralization, were 2–15 times greater on UV-treated scaffolds, with more pronounced enhancement on acid-etched scaffolds compared to that on the original scaffolds. These effects of UV treatment were associated with a significant reduction in atomic carbon on the Ti microfiber surfaces. In conclusion, UV treatment of Ti microfiber scaffolds tunes their physicochemical properties and effectively enhances the attachment and function of osteoblasts, proposing a new strategy for bone engineering.

Transcriptome Analysis of the Ammonia-Oxidizing Bacterium Nitrosomonas mobilis Ms1 Reveals Division of Labor between Aggregates and Free-living Cells.

Bacteria change their metabolic states to increase survival by forming aggregates. Ammonia-oxidizing bacteria also form aggregates in response to environmental stresses. Nitrosomonas mobilis, an ammonia-oxidizing bacterium with high stress tolerance, often forms aggregates mainly in wastewater treatment systems. Despite the high frequency of aggregate formation by N. mobilis, its relationship with survival currently remains unclear. In the present study, aggregates were formed in the late stage of culture with the accumulation of nitrite as a growth inhibitor. To clarify the significance of aggregate formation in N. mobilis Ms1, a transcriptome analysis was performed. Comparisons of the early and late stages of culture revealed that the expression of stress response genes (chaperones and proteases) increased in the early stage. Aggregate formation may lead to stress avoidance because stress response genes were not up-regulated in the late stage of culture during which aggregates formed. Furthermore, comparisons of free-living cells with aggregates in the early stage of culture showed differences in gene expression related to biosynthesis (ATP synthase and ribosomal proteins) and motility and adhesion (flagella, pilus, and chemotaxis). Biosynthesis genes for growth were up-regulated in free-living cells, while motility and adhesion genes for adaptation were up-regulated in aggregates. These results indicate that N. mobilis Ms1 cells adapt to an unfavorable environment and grow through the division of labor between aggregates and free-living cells.

Simulation of Methylmercury Content and SRB Methylation in Phragmites australis Soil Under Different Salinity Conditions

The MeHg content in the reed wetland soil of Liaohe was studied by the indoor-simulated constant temperature culture method. Under different aerobic and anaerobic conditions, the flooding salinity (CK, 0.5%, 1.0%, 1.5%, 2.0%) changes relationship whether SRB plays a leading role in the formation of MeHg. The results showed that under aerobic conditions, the content of MeHg in the surface layer and the bottom layer showed a trend of decreasing first and then increasing with the increase of culture time. Both of them have a lower MeHg content when the flooding salinity is 2.0%. The number of SRB bacteria showed a trend of “upgrading-depleting” with the increase of flooding salinity. Under anaerobic conditions, the MeHg content of surface and bottom soil changed slowly in the early stage of culture (the first 10 days), and the MeHg content increased rapidly after 15 days of culture, and decreased significantly on the 25th day. The number of SRB bacteria showed a trend of “depleting-upgrading” as the flooding salinity increased. Linear fitting showed that there was no obvious linear relationship between the change of MeHg content in soil and the number of SRB bacteria, and other microorganisms may play a role in methylation of mercury. Under anaerobic conditions, MeHg content in surface soil was significantly positively correlated with organic matter (p < 0.01), but negatively correlated with total mercury (p < 0.05). The mercury methylation process is affected by many environmental factors, and the mechanism of mercury methylation in different environments is different.

Effect of Organic and Chemical Fertilizer Application on Growth, Yield, and Soil Biochemical Properties of Landrace &amp;lt;i&amp;gt;Brassica napus&amp;lt;/i&amp;gt; L. Leaf-and-Stem Vegetable and Landrace (Norabona)

Norabona is generally cultivated in Japan under management systems that use chemical fertilizers and synthetic chemical pesticides. However, the continuous use of these fertilizers and pesticides damages the soil environment and reduces the number of soil microorganisms. There has been little research investigating the effect of organic and chemical fertilizer applications on soil biochemistry and the growth and yield of norabona. In this study, we investigated the effect of organic and chemical fertilizer application on these factors during the norabona growing season from September 2019 to May 2020. Leaf length, shoot height, and shoot width were significantly higher under organic fertilizer management in the early stage of cultivation (in March) than under chemical fertilizer management. However, there was no significant difference between treatments for these growth parameters in later months, nor for any other parameters. Soil TN, and TP contents were significantly higher in the organic fertilizer treatment after harvest than prior to cultivation or after the chemical fertilizer treatment. In addition, soil TC, and volumetric water content were significantly higher in the organic fertilizer treatment than in chemical fertilizer treatment. The higher TC, TN, and C/N ratio in organic fertilizer treated soil appeared to increase the bacterial biomass, leading to enhanced nutrient circulation via N and P circulation activity, producing a rich soil environment with active soil microorganisms.

НОВЫЕ ДАННЫЕ О СОСТАВЕ СТАДА У НАСЕЛЕНИЯ СРЕДНЕГО — ПОЗДНЕГО БРОНЗОВОГО ВЕКА В СЕВЕРО-ЗАПАДНОМ КРЫМУ

The paper deals with the results of an archeo-zoological study of bone materials from a num- ber of newly discovered settlements in Northwestern Crimea (Tarkhankut-Н2, Tarkhankut-Н8, Tarkhankut-18 and Tarkhankut-22а). According to the available archaeological evidence and ra- diocarbon determinations obtained on bone remains from cultural layers and semi-closed assem- blages, the materials in question belong to the early stage of the Babino culture (Tarkhankut-18, Tarkhankut-22а) and to the Sabatinovka (Tarkhankut-Н2, Tarkhankut-22а) and Belozerka (Tarkhankut-Н8, Tarkhankut-18) cultures. In addition, the Tarkhankut-22а settlement yielded some Early Iron Age materials. Altogether, 1211 bones were studied, and 284 of them proved to be identifiable. An attempt is made to consider the herd composition in its dynamics: during the final stage of the Middle Bronze Age the herd was heavily dominated by cattle, followed by small ru- minants, while the percentage of horse remains was insignificant. The Late Bronze Age witnessed an increase in the numbers of small ruminants, and the presence of horse became perceptible in the final of the Bronze Age. The authors compare their data with the information available for the Middle and Late Bronze Age settlements of Crimea and the synchronous cultures of the North Black Sea region. The analyzed materials point to the existence of pastoral stockbreeding with stone architecture represented by livestock enclosures.

The &lt;i&gt;Parachlorella&lt;/i&gt; Genome and Transcriptome Endorse Active RWP-RK, Meiosis and Flagellar Genes in Trebouxiophycean Algae

The genus Chlorella is a well-known member of the green algal class Trebouxiophyceae, which is characterized by an immotile and asexual life cycle. Here, we performed an analysis of the whole genome and transcriptome of Parachlorella kessleri NIES-2152 with emphasis on the evolution of meiosis and the flagellar proteins. The Parachlorella transcriptomic data showed that the MID-related RWP-RK genes and meiosis-specific and flagellar proteins were expressed; at the transcriptional level, the DNA repair protein RAD50 was upregulated in the stationary phase, with four-fold more reads per kilobase of transcript per million mapped reads (RPKM) compared with the early stage of culture. In contrast, radial spoke protein genes were down-regulated in the stationary phase. These results suggest that genes for meiotic and flagellar proteins are culture stage-dependent and retain their functions. We presume that the algae lost some of the genes for meiosis and the flagella during asexual evolution, but other genes still possess biological functions other than those related to the flagellum and meiosis.

Identification of a Novel Transcription Factor TP05746 Involved in Regulating the Production of Plant-Biomass-Degrading Enzymes in Talaromyces pinophilus.

Limited information on transcription factor (TF)-mediated regulation exists for most filamentous fungi, specifically for regulation of the production of plant-biomass-degrading enzymes (PBDEs). The filamentous fungus, Talaromyces pinophilus, can secrete integrative cellulolytic and amylolytic enzymes, suggesting a promising application in biotechnology. In the present study, the regulatory roles of a Zn2Cys6 protein, TP05746, were investigated in T. pinophilus through the use of biochemical, microbiological and omics techniques. Deletion of the gene TP05746 in T. pinophilus led to a 149.6% increase in soluble-starch-degrading enzyme (SSDE) production at day one of soluble starch induction but an approximately 30% decrease at days 2 to 4 compared with the parental strain ΔTpKu70. In contrast, the T. pinophilus mutant ΔTP05746 exhibited a 136.8–240.0% increase in raw-starch-degrading enzyme (RSDE) production, as well as a 90.3 to 519.1% increase in cellulase and xylanase production following induction by culturing on wheat bran plus Avicel, relative to that exhibited by ΔTpKu70. Additionally, the mutant ΔTP05746 exhibited accelerated mycelial growth at the early stage of cultivation and decreased conidiation. Transcriptomic profiling and real-time quantitative reverse transcription-PCR (RT-qPCR) analyses revealed that TP05746 dynamically regulated the expression of genes encoding major PBDEs and their regulatory genes, as well as fungal development-regulated genes. Furthermore, in vitro binding experiments confirmed that TP05746 bound to the promoter regions of the genes described above. These results will contribute to our understanding of the regulatory mechanism of PBDE genes and provide a promising target for genetic engineering for improved PBDE production in filamentous fungi.

Effects of polymer materials on soil physicochemical properties and bacterial community structure under drip irrigation

Polymer materials have the effect of retaining water and increasing yield in irrigated agricultural systems. However, little is known about how polymer materials affect soil properties. The purpose of this study was to evaluate the effect of humic acid (H) (an alkaline extract of cottonseed meal), modified polymer (P) (a mixture of anionic polyacrylamide, polyvinyl alcohol, and manganese sulfate), and composite polymer (HP) (1:1 mixture of H and P) on soil physicochemical properties and structural diversity of bacterial community in soils with/without plants under drip irrigation. The results demonstrated that the application of polymer materials decreased soil bulk density, and increased soil moisture content. Compared with the control in soil with plants (CK), the H and P treatments decreased soil bulk density (BD) by 0.09 g·cm−3 and 0.08 g·cm−3, respectively (p < .05) after 90 days of culture. Meanwhile, the H, P, and HP treatments increased soil moisture content (MC) by 10.6%, 33.9%, and 16.5%, respectively (p < .05). For soil without plants, the P-NP treatment decreased BD by 0.18 g·cm−3 (p < .05) after 90 days of culture. Although there were differences in soil pH and conductivity (EC) between treatments, they were not significant. As the culture time was prolonged, the relative abundance of soil Latescibacteria in soil without plants significantly increased compared with that in soil with plants. Principal component analysis (PCA) showed that the effects of polymer material application on soil microbes were not significantly different at the early stage of culture. However, there was significant (in soil with plants, r = 0.540, p = .004) or extremely significant difference (in soil without plants, r = 0.333, p = .045) between treatments after 90 days of culture. Among them, the difference between P treatment and CK was the most significant. Further analysis showed that the P treatment increased relative abundances of soil Acidobacteria, Gemmatimonadetes, and Nitrospirae by 20.6%, 42.8%, and 31.1% compared with CK, respectively (p < .05); and the P-NP treatment increased relative abundances of Gemmatimonadetes, Chloroflexi, and Latescibacteria by 60.5%, 35.7%, and 29.3% (p < .05) compared with CK-NP, respectively. The redundant analysis (RDA) showed that the application of polymer materials could improve the structure diversity of soil bacterial communities by increasing soil MC and regulating C/N ratio, regardless of soil with/without plants.

Chloropicrin fumigation alters the soil phosphorus and the composition of the encoding alkaline phosphatase PhoD gene microbial community

The transformation of phosphorus (P) compounds in soil depends largely on soil microbial communities and is sensitive to agricultural practices. However, the effects of soil fumigation on soil P, and microbes involved in P transformation, are unknown. Our results showed that chloropicrin (CP) fumigation significantly increased the available-P, Leached-P and active-P fractionation (inorganic P extracted from H2O, NaHCO3 and NaOH) in Shangdong and Miyun soils in the early stages of culture, while soil alkaline phosphatase (ALP) activity and phoD gene abundance decreased significantly. Leached-P in fumigated soil was positively correlated with increased active-P fractionation, indicating that it was an important source of soil Leached-P after fumigation. The changes in P-fractionation, Leached-P and ALP after fumigation were also significantly correlated with the composition of the microbial communities. CP fumigation briefly stimulated an increase in the abundance and diversity of phoD-harboring microbial communities and promoted the mineralization process of soil P. PICRUSt metagenomic analysis showed an increase in the relative abundance of microorganisms with involved in carbohydrate/lipid transport and metabolism functions after fumigation. These results suggest CP fumigation altered soil P transformation and phoD-harboring microbes that might lead to an increased risk of P enrichment in waterways.

Effects of graphene oxide on white poplar X ASPEN (Populus alba x Populus tremula) hybrid microsprouts at various growth stages

Graphene-like materials are promising in biotechnology as antimicrobial agents and plant growth stimulants. We investigated the effects of graphene oxide nanoparticles in the composition of the MS0 medium on microclonal seedlings of the poplar and aspen hybrid. In the early stages of cultivation, graphene oxide improved microclone survival by 6–20%. However, then, along with some stimulation, negative effects were observed, manifested in a decrease in the development of the root system and a decrease in the thickness of the leaf lamina and the diameter of the stem. Our results indicate the promise of using graphene oxide in the early stages of the development of microclones of white poplar and aspen hybrids.

Lactobacillus fermentum and/or ferulic acid improved the immune responses, antioxidative defence and resistance against Aeromonas hydrophila in common carp (Cyprinus carpio) fingerlings

This study investigates the possible effects of using Lactobacillus fermentum (LF) and/or ferulic acid (FA) in common carp (Cyprinus carpio) on some immunological parameters as well as resistance against Aeromonas hydrophila. Four diets were prepared including control diet and three diets supplemented with LF (108 CFU/g), FA (100 mg kg−1) or LF + FA (108 CFU/g + 100 mg kg−1). After 8 weeks, fish fed LF or/and FA had significantly higher final body weight, weight gain, and specific growth rate when compared to control group (P < 0.05). The feed conversion ratio of fish fed LF or/and FA were noticeably lower than control (P < 0.05). No alterations were observed in case of haematological parameters except red blood cells (RBCs), white blood cells (WBCs), hemoglobin (Hb), and hematocrit (HCT) which were significantly (P < 0.05) increased in fish fed FA or those fed both LF and FA. Also, the WBCs of fish treated with LF or/and FA were noticeably higher than control (P < 0.05). Feeding on LF and FA notably increased the serum total protein and albumin levels (P < 0.05). The serum respiratory burst and lysozyme activity were also enhanced (P < 0.05) in fish fed both LF or/and FA. In addition, evaluation of the serum antioxidant enzymes (catalase, glutathione peroxidase (GPX), and superoxide dismutase (SOD)) activity showed significant (P < 0.05) increase in fish fed FA or both LF and FA as compared to the control. Fish fed LF and FA supplemented diet had highest survival rate after experimental challenge with pathogenic A. hydrophila. The obtained results revealed that LF and/or FA can be used as beneficial feed additive to improve the immune responses and disease resistance in early stages of common carp culture.

The Absence of Tryptase Mcpt6 Causes Elevated Cellular Stress in Response to Modulation of the Histone Acetylation Status in Mast Cells.

Mast cells contain large amounts of proteases stored within their secretory granules. Previously we showed that one of these proteases, tryptase, in addition to its location within granules, can also be found within the mast cell nucleus, where it has the capacity to affect the acetylation profile of nucleosomal core histones in aging cells. Based on this notion, and on the known sensitivity of mast cells to modulation of histone acetylation, we here asked whether tryptase could impact on the responses against cellular stress caused by disturbed histone acetylation status. To address this, wild-type and tryptase-deficient (Mcpt6−/−) mast cells were subjected to cell stress caused by trichostatin A (TSA), a histone deacetylase inhibitor. Wild-type and Mcpt6−/− mast cells were equally sensitive to TSA at an early stage of culture (~8 weeks). However, in aging mast cells (>50 weeks), tryptase-deficiency led to increased sensitivity to cell death. To address the underlying mechanism, we assessed effects of tryptase deficiency on the expression of markers for proliferation and cell stress. These analyses revealed aberrant regulation of thioredoxin, thioredoxin reductase, glutaredoxin, and glutathione reductase, as well as blunted upregulation of ribonucleotide reductase subunit R2 in response to TSA in aging cells. Moreover, the absence of tryptase led to increased expression of Psme4/PA200, a proteasome variant involved in the processing of acetylated core histones. Altogether, this study identifies a novel role for tryptase in regulating the manifestations of cell stress in aging mast cells.

Germination and survival of Aechmea gamosepala on different substrates

Abstract The State of Parana is one of the Brazilian regions with the highest incidence of extraction of bromeliads, among the factors for the occurrence of this situation, they highlight a great demand for this species as ornamental flowers and the easy access to forests, few cultivation ponds and difficulties for farmers in cultivation processes, especially in the early stages of cultivation. The species Aechmea gamosepala Wittmack stands out among the species of this group of ornamental plants (Bromeliaceae) with strong commercial appeal and consequently of extraction. Given this context, a study was carried out to evaluate the germination and survival of seeds of the species. Five alternative types of substrates easily accessible to farmers were tested: coconut husk fiber, Plantmax®, earthworm humus, humidified Pinnus husk and sieved agricultural land. The experiment was conducted in a completely randomized design with 4 replications and 25 seeds per experimental unit at room temperature in a greenhouse, with sprinkler irrigation for a period of 90 days. The mean time for the onset of germination was 6.95 days, averaging 74.8% at the peak of germination, but there were no significant statistical differences between the treatments. For the seedling survival, humidified Pinnus husk substrates (70.8%) and coconut fiber house (80.4%) proved to be the best substrates for Aechmea gamosepala Wittmack.

A new understanding: Gene expression, cell characteristic and antioxidant enzymes of Zygosaccharomyces rouxii under the D-fructose regulation

Zygosaccharomyces rouxii is a well-known salt-tolerant yeast. In our previous study, it was interesting that Z. rouxii could produce higher levels of 4-hydroxy-2, 5-dimethyl-3(2 H)-furanone in 120 g/L D-fructose and 180 g/L NaCl involved YPD medium at 5 d. In order to explore the resistance and furanone production mechanisms of Z. rouxii under D-fructose regulation, a comparative transcriptomics method in Z. rouxii was to set to find differentially expressed genes, the physiological and biochemical indexes (growth and cell morphology, lipid peroxidation and relative electrical conductivity, the antioxidant enzymes activity), and the expression of oxidoreductase activity genes. The results indicated that a larger number of different expressed genes at transcriptome analysis, such as the series antioxidant enzymes were related to the resistance characteristics. Research had confirmed that the living cell numbers and cell areas of D-fructose regulation group were significantly lower than the controls at the initial stage, while those higher than of the controls at the late stage. During the fermentation period, the lipid peroxidation and the relative electrical conductivity of the yeast cell membrane were increased. And also the D-fructose regulation group present lower inhibition superoxide anion ability. The activity of CAT in the D-fructose regulation group was always higher than that of the control group. Only the activity of GSH-Px was found to be significantly increased at 1 d except for other enzymes activities. Most of the oxidoreductase activity genes, such as especially the GSH-Px gene under D-fructose regulation conditions were expressed at higher levels than those of control groups. Combining the levels of transcription and enzymes activity data, those could understand that exogenous D-fructose had a stress effect on Z. rouxii at the early stage of culture. With the fermentation time progress, it was no longer a stressor substance for the Z. rouxii, and changed the nutrient to promote growth of Z. rouxii in the later stages. During the whole process, GSH-Px was the main defense enzyme and CAT was the sustained defense enzyme. Therefore, the experimental results might provide effective mechanisms in Z. rouxii for practical application of furanone production in the industry under exogenous D-fructose regulation.

Estimation of Ground Surface and Accuracy Assessments of Growth Parameters for a Sweet Potato Community in Ridge Cultivation

There are only a few studies that have been made on accuracy assessments of Leaf Area Index (LAI) and biomass estimation using three-dimensional (3D) models generated by structure from motion (SfM) image processing. In this study, sweet potato was grown with different amounts of nitrogen fertilization in ridge cultivation at an experimental farm. Three-dimensional dense point cloud models were constructed from a series of two-dimensional (2D) color images measured by a small unmanned aerial vehicle (UAV) paired with SfM image processing. Although it was in the early stage of cultivation, a complex ground surface model for ridge cultivation with vegetation was generated, and the uneven ground surface could be estimated with an accuracy of 1.4 cm. Furthermore, in order to accurately estimate growth parameters from the early growth to the harvest period, a 3D model was constructed using a root mean square error (RMSE) of 3.3 cm for plant height estimation. By using a color index, voxel models were generated and LAIs were estimated using a regression model with an RMSE accuracy of 0.123. Further, regression models were used to estimate above-ground and below-ground biomass, or tuberous root weights, based on estimated LAIs.

Pottery from the Defensive Moat of “Citadel” Site of Dykyi Sad Hillfort

The article deals with pottery from the filling of the defensive moat of “Citadel” site of the hillfort Dykyi Sad. The moat has a shape of a large arc that is elongated along the line South-East–North-West. The length of the investigated part is 1.30 m, width – 5.0 m, depth – 3.0-3.10 m. There were identified more than twenty-two thousand fragments of pottery, belonging to about 2500 vessels of various purposes. The moat filling can be divided into three levels: – upper level (thickness 1.0 m), in which the expressionless ceramics were fixed (about 1000). – middle level (thickness 1.0 m), in which the saturation of ceramic fragments increases (approximately 10000). – lower level (thickness 1.0 m), in which the number of fragments of ceramic ware is practically similar to the middle level (about 11000). There were selected 165 complete and partly preserved vessels from the middle (81 fragment) and lower filling levels (84 fragments), suitable for formal classification. The subject of the study is moulded ware from the middle and lower levels of moat filling. All material is divided into two groups: closed (120 fragments) and open shapes (41 fragment). Each group is divided into types. Pottery of closed group consists of pots, jars, cups, ladle, large pots. The open group wares are bowls, cups, braziers, riddles. Among the forms there were specified types due to the shape of neck and rim. According to the type there have been classified variations within body shapes. A separate, special, group is composed with non-stan¬dard (miniature) dishes, which can be considered as for cult purposes (4 fragments). It was found that among closed group dominated squat cups (32,5 %) and pots with arc-like neck (14,2 %); the lowest number is represented by elongated cups (4,2 %) and pots with conical neck (1.6 %). Among the dishes of open group there are bowls (36.6 %), and cups (31,7 %); the least represented are flat braziers with segmental-cut body form (7,3 %), bowls with elongated straight neck (7.3 %), riddles (7.3 %) and flat braziers with a small rim (4.9 %). Among the miniature (cult) dishes there are two jars (50 %), a cup (25 %) and a small pot (25 %). Pottery from the moat filling relates to the ceramics fixated in the “Citadel” premises of the hillfort Dykyi Sad and is found among mat¬erials of well-known settlements and burial grounds of Bilozerka culture period of the North-Western Pontic region. In the cultural and chronological aspect the ceramics of lower level moat filling of the “Citadel”, there are mostly fragments refering to the early stage of Bilozerka culture and early stage of the settlement occurence (end of 13th – beginning of 12th c. BC), and the ceramics of middle level relates to the main period of the hillfort existence (12th – 11th c. BC).

Highly selective isolation and characterization of Lipomyces starkeyi mutants with increased production of triacylglycerol.

The oleaginous yeast Lipomyces starkeyi is an attractive organism for the industrial production of lipids; however, the amount of lipid produced by wild-type L. starkeyi is insufficient. The study aims to obtain L. starkeyi mutants that rapidly accumulate large amounts of triacylglycerol (TAG). Mutagenized yeast cells at the early stages of cultivation were subjected to Percoll density gradient centrifugation; cells with increased production of TAG were expected to be enriched in the resultant upper fraction because of their lower density. Among 120 candidates from the upper fractions, five mutants were isolated that accumulated higher amounts of TAG. Moreover, when omitting cells with mucoid colony morphology, 11 objective mutants from 11 candidates from the upper fraction were effectively (100%) isolated. Of total 16 mutants obtained, detailed characterization of five mutants was performed to reveal that five mutants achieved about 1.5-2.0 times TAG concentration (4.7-6.0g/L) as compared with the wild-type strain (3.6g/L) at day 5. Among these five mutants, strain E15 was the best for industrial use because only strain E15 showed significantly higher TAG concentration as well as significantly higher degree of lipid to glucose and biomass to glucose yields than the wild-type strain. Thus, Percoll density gradient centrifugation is an effective method to isolate mutant cells that rapidly accumulate large amounts of TAG. It is expected that by repeating this procedure as part of a yeast-breeding program, L. starkeyi mutants suitable for industrial lipid production can be easily and effectively obtained.

3D printed porous Ti6Al4V cage: Effects of additive angle on surface properties and biocompatibility; bone ingrowth in Beagle tibia model

Similar mechanical properties to native bone, excellent biocompatibility and osseointegration are key factors for interbody fusion cage. In the present study, porous Ti6Al4V cage with computer designed macro- and micro-architecture was reproduced by selective laser melting (SLM). Dense Ti6Al4V discs with six different angles were fabricated to investigate the effects of additive angle (inevitable in SLM manufacturing) on surface properties and biocompatibility. As angle increased, both surface roughness and hydrophobicity increased due to increases in the number of unmelted metallic particles. In the early stage of cell culture, more elongated MC3T3E1 cells were observed at higher additive angle, but cell adhesion and proliferation with different angle did not show statistically differences. Bullet-shaped Ti6Al4V cage with opened window, porosity of 70% and pore size of 600 μm revealed elastic modulus of 0.51 ± 0.04 GPa and ultimate compressive strength of 94.7 MPa at 0.29% strain, which is comparable with the commercialized polyetheretherketone cage. In vivo evaluation of Beagle tibia model through histological analysis showed that the cylindrical porous body with same porosity and pore size achieved similar bone ingrowth as commercialized porous tantalum implant at 4 and 12 weeks, indicating SLM fabricated porous Ti6Al4V cage holds great potential for spinal fusion.

Degradation of antifungal anthraquinone compounds is a probable physiological role of DyP secreted by Bjerkandera adusta

Alizarin is an anti-fungal compound produced by the plant, Rubia tinctorum. The parasitic fungus Bjerkandera adusta Dec 1 was cultured in potato dextrose (PD) medium with or without alizarin. Alizarin was a good substrate for the dye-decolorizing peroxidase (DyP) from B. adusta Dec 1 and hampered B. adusta growth at the early stage of plate culture. During liquid shaking culture, DyP activity in cultures supplemented with 100 μM alizarin was greater than that in controls cultured without alizarin. In particular, DyP activity per dry cell mass increased approximately 3.5-, 3.1-, and 2.9-fold at 24, 30, and 36 h after inoculation, respectively, compared with control cultures. These data suggest that alizarin stimulates the expression of DyP. Interestingly, alizarin rapidly decomposed at an early stage in culture (24–42 h) in PD medium supplemented with 100 μM alizarin. Thus, alizarin appears to induce DyP expression in B. adusta Dec 1, and this DyP, in turn, rapidly degrades alizarin. Collectively, our findings suggest that the physiological role of DyP is to degrade antifungal compounds produced by plants.