Early Stages Of Culture Research Articles

Growth of Bifidobacterium pseudocatenulatum in medium containing N-acetylsucrosamine: enzyme that induces the growth of this bacterium via degradation of this disaccharide.

Bifidobacterium pseudocatenulatum grows well in the early stages of cultivation in medium containing sucrose (Suc), whereas its growth in medium containing the analogue disaccharide N-acetylsucrosamine (SucNAc) tends to exhibit a considerable delay. To elucidate the cause of this phenomenon, we investigated the proliferation pattern of B. pseudocatenulatum in medium containing D-glucose (Glc) and SucNAc and identified the enzyme that degrades this disaccharide. We found that B. pseudocatenulatum initially proliferates by assimilating Glc, with subsequent growth based on SucNAc assimilation depending on production of the β-fructofuranosidase, which can hydrolyze SucNAc, after Glc is completely consumed. Thus, B. pseudocatenulatum exhibited a diauxic growth pattern in medium containing Glc and SucNAc. In contrast, when cultured in medium containing Glc and Suc, B. pseudocatenulatum initially grew by degrading Suc via the phosphorolysis activity of Suc phosphorylase, which did not react to SucNAc. These observations indicate that B. pseudocatenulatum proliferates by assimilating Suc and SucNAc via different pathways. The β-fructofuranosidase of B. pseudocatenulatum exhibited higher hydrolytic activity against several naturally occurring Suc-based tri- or tetrasaccharides than against Suc, suggesting that this enzyme actively catabolizes oligosaccharides other than Suc.

Ketamine used in the therapy of depressive disorders impacts protein profile, proliferation rate, and phagocytosis resistance of enterococci

Abstract Introduction A low concentration of ketamine is used to cause an anti-depressive effect. The mechanism of ketamine's action in depression is believed to result, among others, from its anti-inflammatory activity. Despite the fact that only high concentrations of ketamine inhibit bacterial growth, it is clear that even a sub-inhibitory concentration of chemicals may change bacterial properties. Considering the above, in the current study we aimed to evaluate the in vitro influence of ketamine on proliferation of enterococci and their interactions with monocytes. Materials and Methods The studied strains were isolated as etiological agents of infection at Medical University of Gdansk. The proliferation and metabolic activity were determined using the FACSVerse flow cytometer after addition of CFDA-SE to bacterial suspension. For the determination of phagocytosis resistance, THP-1 human monocytes cell line was used. Suspension of monocytes which engulfed CFDA-SE–stained bacteria was then stained with propidium iodide to evaluate cytotoxicity of enterococci. Results The result of the study showed unexpected response of bacterial cells to ketamine at an early stage of culture. In 57.7% of strains, both proliferation rate and metabolic activity were boosted. This group of strains was also less susceptible to phagocytosis than in culture without ketamine. Different response of isolates to ketamine was also visible in changes of proteins’ profile determined by MALDI-TOF. Conclusions The analysis of bacteria at an early stage in the growth curve demonstrated the bacterial diversity in response to ketamine and let us set the hypothesis that microbiome susceptibility to ketamine may be one of the elements which should be taken into consideration when planning the successful pharmacotherapy of depression

Бронзовый наконечник стрелы из позвонка мужчины, погребенного в тасмолинском кургане Койтас

In 2011 excavations of the Koytas emergency burial mound in Central Kazakhstan (in Karkaraly district of Karaganda region) recorded features of a burial structure characteristic of the early stage of the Tasmola culture. It turned out that the studied site had been repeatedly exposed to looting. The mound contained a dromos leading to a burial mound. The parameters of the embankment (diameter 22.5 m, height 2.5 m) are reconstructed, a circular crepice and a ditch are identified. In the burial chamber, some bones from a human skeleton have been preserved. The body of the only surviving vertebra, belonging to the lower thoracic region, was found to contain a trapped metal petiole arrowhead. As previous studies have shown (Tur et al., 2016), the severe wound did not immediately result in death. The signs of healing are clearly visible. This represents a very rare case for the ancient era. X-ray fluorescence analysis showed that the arrowhead was made of a quality bronze alloy, which contributed to a certain extent to prolonging the man’s life. According to the funerary rites and the features of the arrowhead, as well as on the basis of radiocarbon dates, the Koytas barrow dates back to the period of the 8th to 6th centuries B.C.

Synthetic Data Augmentation of Tomato Plant Leaf using Meta Intelligent Generative Adversarial Network: Milgan

Agriculture is one of the most famous case studies in deep learning. Most researchers want to detect different diseases at the early stages of cultivation to save the farmer's economy. The deep learning technique needs more data to develop an accurate system. Researchers generated more synthetic data using basic image operations in traditional approaches, but these approaches are more complicated and expensive. In deep learning and computer vision, the system's accuracy is the crucial component for deciding the system's efficiency. The model's precision is based on the image's size and quality. Getting many images from the real-world environment in medicine and agriculture is difficult. The image augmentation technique helps the system generate more images that can replicate the physical circumstances by performing various operations. It also prevents overfitting, especially when the system has fewer images than required. Few researchers experimented using CNN and simple Generative Adversarial Network (GAN), but these approaches create images with more noise. The proposed research aims to develop more data using a Meta approach. The images are processed using kernel filters. Different geometric transformations are passed as input to the enhanced GANs to reduce the noise and create more fake images using latent points, acting as weights in the neural networks. The proposed system uses random sampling techniques, passes a few processed images to the generator component of GAN, and the system uses a discriminator component to classify the synthetic data created by the Meta-Learning Approach.

HOARD OF ANTONINIANI FROM THE SOUTHERN POBUZHIA

In autumn 2009, 33 whole and 3 fragmented аntoniniani were discovered and collected within the object of the Late Roman Age - Pereginka (Balakiri)2.
 According to available information, mostly the coins were concentrated in a limited area with sides of about 15 by 10 meters, and only a few ones were spread by plowing equipment at a distance of up to 30 meters. The distribution of coins and the general state of preservation suggests the collected аntoniniani may have been a part of the deposit, which lay at a depth of 25-40 cm, and was made on the day surface due to the use of more powerful attachments that is used in recent years in the cultivation of agricultural land.
 In the following years, another five whole and three fragmented аntoniniani were found. The total number of discovered coins is 44 copies.
 Chronologically, the complex is determined by the lower date – 238 A.D. – the issues of Gordian III: «IOVI CONCERVATORI» and «PROVIDENTIA AVG»; and the upper – 251-253 A.D. – the issues of Trebonianus Gallus «PIETAS AVGG» and Voluzian «PAX AVGG». The chronological framework of the complex's emissions is a relatively short period – about 15 years.
 The complex became the first fixed and attributed deposit of аntoniniani in the forest-steppe zone of South-Eastern Europe and it is extremely important for understanding the processes of formation of Chernyakhiv culture, as well as, further analysis and development of socio-economic relations of the Roman Empire and «Barbarian» communities in the east of the Danube Limes line.
 At the same time, the limited source base makes no sense to draw any conclusions about the ethnicity or social status of the deposit holder, as well as about possible ways of entering аntoniniani from the complex Perehinka (Balakiri) 2 on the territory of the Ukrainian Forest-Steppe. To receive conditions close to reality it is possible under the condition of detectiono of the closed complex (or its additional inspection) at carrying out full-fledged archeological researches of the given object.
 Undoubtedly, one can declare working hypotheses: the аntoniniani came here as prey captured by the «barbarians» during the fighting of 247-251 A.D.; or as a part of the ransom payments promised by Trebonianus Gallus to the «barbaric» tribal or military elite. It can be assumed that the аntoniniani belonged to one of the soldiers of the legions stationed in Moesia, who rebelled and joined the coalition of «barbarian» tribes during the «Scythian» wars, we can appeal to the testimony of Zosimus (Zos., I, 28) about a successful campaign of the Roman units in 253 A.D. those were led by Aemilian, during which not only the detachments of «barbarians» led by Kniva were defeated, but also a successful raid was carried out in their land. It is possible to consider assumptions that the complex could be a part of monetary maintenance of the serviceman or the veteran of regular or federal divisions… But – concerning each of the above-stated hypotheses it will be possible to find and express equal argumentation «for» and «against». As with the typology, the deposit is classified as a single-nominal or mixed treasure, as not only denarii but also sesterces and provincial issues are found on the site.
 We should also pay attention to another key point. The оbject Perehinka (Balakiri) 2, territorially belonging to the upper reaches of the Southern Bug, is located in a region characterized not only by high density and extreme saturation of monuments those belong to the Chernyakhiv culture, but also as one of the main areas its formation. It is here the objects represented by artifacts attesting to their belonging to the early stage of the Chernyakhiv culture are concentrated: Ruzhychanka, Cherneliv-Rusky, Berezhanka, Lepesivka, Luka-Vrublivetska, Chankiv I, and others.
 If we take into account the chronology of the early stage of Chernyakhiv culture- the so-called «ruzhychanka phase» (according to E. Gorokhovsky) – 230-270 A.D.; the «first phase» (according to I. Bazhan and A. Gay), which are generally synchronized with phase C1 of the Central-European chronological system – 220-260 A.D. – then the complex of аntoniniani from Pereginka (Balakiri)2 can be one of its chronological benchmarks, as an example, and аntoniniani Valerian and Trajan Decius (for Gerenia Etruscilla) in one of the burials of the Chernyakhiv necropolis of Talalayevka (Khrystynivskyi district, Cherkasy region).

Enhanced triacylglycerol accumulation in open cultivation of microalgae using an air self-sufficient aerator

To improve energy production cost, it is necessary to operate bioreactors at a deeper depth to increase per unit area production; however, self-shading could be an inhibiting factor. Therefore, it is important to employ a variety of agitators so that microalgae in deep regions can be agitated, allowing sufficient aeration. We aimed to evaluate the effectiveness of a self-sufficient aerator in an open pond cultivation system for a microalga. Three experimental cases with different agitation velocities (high: Casehigh; low: Caselow; no agitation: Casezero) were evaluated. In Caselow, cells grew fastest in the early stage of cultivation due to reduced mechanical shear stress. However, the increased turbidity after 150 h reduced the cell density and increased chlorophyll a content, which could be attributed to low light intensity. The maximum TAG content was achieved in Casehigh. The findings suggest that strong agitation using an aerator can promote TAG accumulation.

The Expression of CD28 and Its Synergism on the Immune Response of Flounder (Paralichthys olivaceus) to Thymus-Dependent Antigen.

CD28 is well known as a critical T-cell costimulatory receptor involved in T cell activation by binding to its ligands. In this study, CD28 was cloned, and its expression profiles were characterized in flounder (Paralichthys olivaceus); variations of CD28+ cells after being stimulated with different types of antigens and the function of the CD28 costimulatory pathway on T-cell activation were investigated in vitro. fCD28 consists of four exons and three introns, and the full-length cDNA of fCD28 was 675-bp encoded 224 amino acids. The conserved motif (121TFPPPF126) binding to the CD80/86 ligand exists in the Ig-superfamily homology domain. The high expression of fCD28 is in gills, PBLs, head kidney, and spleen. CD28+ cells were co-localized with CD4+ T lymphocytes but not on IgM+ B lymphocyte cells. Moreover, the expression of CD28 was significantly varied in flounder after being stimulated by keyhole limpet hemocyanin (KLH) at both the transcriptional and cellular levels, while no significant differences were observed between lipopolysaccharide (LPS) stimulation and the control group. Notably, treatment of PBLs cultured in vitro with CD28 molecule-specific antibody (anti-CD28 Abs) and PHA produced more cell colonies and stimulated the proliferation of cultured leukocytes compared to PHA stimulation alone and the control group, and a higher level of IL-2 was detected in the culture medium. Meanwhile, anti-CD28 Abs increased the percent of CD28+ cells (10.41 ± 1.35%), CD4+ T lymphocytes (18.32 ± 2.15%), and CD28+/CD4+ double-positive cells (6.24 ± 1.52%). This effect also resulted in significant variations in the genes of cell membrane-bound molecules, cytokines, and related signaling pathways in cultured leukocytes, with significant changes in the genes of interleukin-2 (IL-2) and nuclear factor of activated T cells (NFAT) in the early stages of culture, and the expression of other molecules increased over time. These results proved the localization of the CD28 molecule on T lymphocytes in flounder, and anti-CD28 may act as the B7 ligand involved in T cell activation after antigen stimulation. These data provide a basis for a more in-depth study of the mechanism of the CD28 costimulatory pathway in T cell activation.

RESEARCH OF AMYLOLYTIC ACTIVITY OF CULTURAL LIQUIDS IN BIOTECHNOLOGY OF SYMBIOTIC CROPS МЕDUSOMYCES GISEVII AND ORYZAMYCES INDICI

Based on the literature review found that the natural symbionts Medusomyces gisevii and Oryzamyces indici biotechnology are a valuable objects. It is urgent to find ways to impact on it in order to obtain these or other products of its life activity.
 At present, it is urgent to search for microorganisms producing enzymes, including amylase. One of the most promising in terms of biological objects is a natural microbial symbiont Medusomyces Gisevii (tea fungus) and Oryzamyces Indici, which, thanks to the not identical, microbiological composition and different growing conditions may have a different composition of metabolites. Studies of the amylolytic activity of the culture liquid Medusomyces Gisevii and Oryzamyces indici with different cultivation periods have been carried out. Cultivation of the fungus was carried out in the laboratory according to the classic method. The optimal concentration of sucrose for Medusomyces Gisevii and Oryzamyces Indici biomass growth was set at 5%. Sucrose concentration of 15% and above is not recommended for use due to inhibition of biomass growth with increasing concentration of carbohydrates in the culture medium. It is established that these symbionts start to show the amylolytic activity already on the 10th day of cultivation cultivation in standard nutrient medium, medium supplemented with 10 % milk and serum-based medium and stores it in the course of the experiment (30 days). Over time the amylolytic activity increases. However, the intensity of metabolism of the microorganisms, the criterion of which is the ratio of the total and exogenous amylase is most pronounced in the early stages of cultivation. It was found that the culture medium of polycultures shows high amylolytic activity. This fact allows us to consider the Medusomyces Gisevii and Oryzamyces Indici inoculum as a promising biotechnological raw material source of amylase enzyme.

The dominant microbial metabolic pathway of the petroleum hydrocarbons in the soil of shale gas field: Carbon fixation instead of CO2 emissions

In shale gas mining areas, indigenous microorganisms degrade organic pollutants such as petroleum hydrocarbons into carbon dioxide (CO2) and water (H2O) through aerobic metabolism. A large quantity of CO2 emissions will exacerbate the “Greenhouse effect”. Based on the clean sieved soil and oil-based drilling fluid in the shale gas mining area, this experiment set three concentration gradients (3523 ± 159 mg/kg, 8715 ± 820 mg/kg and 22,031 ± 1533 mg/kg) to treat the soil, and each group was disposed for the same amount of time (63 days). By analyzing the dynamic changes of microbial diversity and the abundance of key functional genes for carbon fixation, the impact of petroleum hydrocarbons on carbon fixation potential was discovered, and the natural attenuation law of petroleum hydrocarbons in contaminated soil was explored. It provided the scientific research basis of ecology for the carbon cycle, carbon allocation, and carbon fixation in microbial remediation of petroleum hydrocarbon contaminated soil. The results obtained indicated the following: i) The removal rate of petroleum hydrocarbons under high-concentration pollution (45.33 ± 3.90%) was significantly lower than low and medium-concentration pollution. The TPH concentration removal rate of each group was the largest in the early stage of culture (1-5d), and there was no significant correlation between the TPH content and the community composition (R2 = 0.0736, P > 0.05). ii) Composition and function of Carbon Fixation associated microbiota were assessed by 16S rRNA sequencing and PICRUSt (phylogenetic investigation of communities by reconstruction of unobserved states) analysis. The main carbon fixation pathway in this study is the reductive citric acid cycle, because there was no shortage of enzymes that can affect subsequent reactions.

Morphological, Histological, and Protein Profiling of Tea Embryo Axis at Early Stage of Culture

Tissue culture is an alternative choice of plant propagation either through somatic embryogenesis or in vitro organogenesis techniques. TRI2025 tea clone has been cultured successfully, however, the scientific information related to morphology, histology, and protein profile at an early event of culturing time has not been reported yet. This study aimed to determine the differences between those pathways, in the context of morphology, histology, and protein profile. The explants were the embryo axis of TRI2025 tea clone cultured on two different induction mediums; somatic embryogenesis and in vitro organogenesis induction medium. The results showed that most of the explants cultured on A medium developed to be a globular-like structure at 11-day after culture (DAC), while all explants cultured on B medium showed the initiation stage of in vitro organogenesis. Histological analysis showed meristem reconstruction at shoot apical meristem (SAM) and root apical meristem (RAM) at 11-DAC at explants cultured on B medium, while explants cultured on A medium showed callusing at 21-DAC. Protein profile analysis using SDS-PAGE showed protein bands of 54 and 81 KDa that only appeared at explants cultured on A medium start from 14-DAC, and those two protein bands thought to be a differentiator at the early stages of the two tissue culture techniques. Thus, these parameters can be used as early detection for plant tissue culture, especially in tea.

Microalgae-based swine wastewater treatment: Strain screening, conditions optimization, physiological activity and biomass potential

Using microalgae to treat swine wastewater (SW) can achieve wastewater purification and biomass recovery at the same time. The algae species suitable for growth in SW were screened in this study, and the response surface combined with the desirability function method was used for multi-objective optimization to obtain high algal biomass and pollutant removal. Chlorophyll fluorescence parameters and biomass composition were analyzed to evaluate the cell physiological activity and its application potential. Chlorella sp. HL was selected as the most suitable species for growth in SW, and after 9 d of cultivation, the maximum specific growth rate and highest algal density were achieved 0.51 d−1 and 2.43 × 107 cells/mL, respectively. In addition, the removal of total phosphate and chemical oxygen demand were reached 69.13% and 72.95%, respectively. The optimum conditions for maximum algal density and highest pollutant removal were determined as the light intensity of 58.73 μmol/m2/s, inoculation density of 5.0 × 106 cells/mL, and a light/dark ratio of 3 using response surface model, and the predicted overall desirability value was 0.96. The potential maximum quantum yield of PSII (Fv/Fm) of Chlorella sp. HL in the early stage of cultivation was 0.60–0.70, while under high light and long photoperiod, the value of Fv/Fm and performance index of Chlorella decreased, trapped and dissipated energy flux per reaction center increased. The higher heating value of 18.25 MJ/kg indicated that the Chlorella cultivated in SW could be a good feedstock for biofuel production.

Effects of Berberine on the Chondrogenic Differentiation of Embryonic Limb Skeletal Progenitors.

IntroductionBerberine (BBR) is an isoquinoline plant alkaloid with demonstrated anti-inflammatory, anti-tumor and immunosuppressive pharmacological properties that functions via multiple signaling pathways and epigenetic modulators. Numerous studies have proposed BBR as a promising therapeutic agent for joint cartilage degeneration, and other connective tissue diseases.Purpose and MethodsThis work aimed to evaluate the effects of BBR on the growth and differentiation of embryonic skeletal progenitors using the limb mesoderm micromass culture assay.ResultsOur findings show that at difference of its apoptotic influence on a variety of tumor tissues, cell death was not induced in skeletal progenitors by the addition of 12 or 25 µM BBR concentration to the culture medium. Morphological and transcriptional analysis revealed dual and opposite effects of BBR treatments on chondrogenesis depending on the stage of differentiation of the cultured progenitors. At early stage of culture, BBR was a potent chondrogenic inhibitor, while chondrogenesis was intensified in treatments at advanced stages of culture. The chondrogenic promoting effect was accompanied by a moderate upregulation of gene markers of prehypertrophic cartilage, including ColXa1, alkaline phosphatase Alpl, Runx2, and Indian Hedgehog Ihh. We further observed a positive transcriptional influence of BBR in the expression of DNA methyltransferase genes, Dnmt1, Dnmt3a and Dnmt3b, suggesting a potential involvement of epigenetic factors in its effects.ConclusionOur study uncovers a new pharmacological influence of BBR in cartilage differentiation that must be taken into account in designing clinical protocols for its employment in the treatment of cartilage degenerative diseases.

Study on the Removal Effect of Salmonella Typhimurium Biofilm

Objective: This research focused on the effects of different disinfectants, preservatives and natural ingredients commonly used in the food industry on the biofilm of Salmonella typhimurium. Methods: By adding different kinds and concentrations of scavengers during the culture process, the influence on the formation of biofilm was determined. The mature biofilm was soaked with various selected scavengers to evaluate the clearance effect under different concentrations of each substance. The metabolic activity of bacteria cells in the biofilm was determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. And the number of viable bacteria was calculated by dilution plate coating method. Results: The disinfectant ethanol (50%), hydrogen peroxide (1%), benzalkonium chloride (0.6%), benzalkonium bromide (0.6%), and natural components carvol, cinnamaldehyde and thyme essential oil (1.0 μL/mL) could effectively inhibit the formation of S. typhimurium CDC3 and S. typhimurium ST34. Disinfectants detergent, hydrogen peroxide, natural ingredients (carvol, cinnamaldehyde and thyme essential oil) showed certain advantages in the removal of mature biofilm. Among them, the most ideal removal efficiency was hydrogen peroxide, which could reach about 90%, the maximum removal rate of detergent could reach 80%, and the essential oils of carvacrol, cinnamaldehyde and thyme could reach about 70%; The disinfectant ethanol, benzalkonium chloride and benzalkonium bromide, as well as the natural components carvol, cinnamaldehyde and thyme essential oil could significantly reduce the cell metabolic activity and the number of viable bacteria in biofilm, the number of viable bacteria in the biofilm could be reduced by 2 to 3 lg values. Conclusions: In contrast with the removal of mature biofilm, the addition of scavenging agent or natural ingredients at the early stage of cultivation made up for a more efficient control of the biofilm.

Relations between Steppe Nomads and Farmers in Eastern Europe and M. Gimbutas’ Migration Concept

After Ukraine had gained independence, artificial problems began to arise in the study of the ancient history of Eastern Europe, especially in the relations of steppe herders and farmers of Polissia. In particular, this applies to the Migration Concept of the American archaeologist M. Gimbutas recognized in Europe. After an unfounded “unscrupulous criticism” of archaeologists – representatives of the “Russian World” – an archaeologist from Moldova V. Dergachev spoke in defense of the objectivity of the 50-year study of this topic by M. Gimbutas in his monograph. Earlier, the Migration Concept of conquering Europe was supported by D. Telehin, Kh. Todorova, N. Ryndina, Ye. Chernykh, and others.Our study confirms the conceptual foundations of the ancient history of relations between nomadic steppes and farmers of Eastern Europe relying on the Migration Concept of M. Gimbutas. On the basis of the sources, protective fortifications of Kukuteny-Trypillia settlements and a large number of arrowheads – weapons of steppe riding archers – were found. In the early, middle, and late stages of Kukuteni-Trypillya culture, three waves of military invasion against European farmers by the Middle-Volga’s, Middle-Stog’s and Novodanyliv’s nomadic tribes of the eastern steppes can be traced. Further research will allow the author to solve the problems of the origins and causes of the disappearance of Kukuten-Trypillia tribes, the problems of burning Trypillia settlements, the fate of Buh-Dniester and East Trypillian tribes.

Dynamic change in bacterial communities in the integrated rice–fish farming system in Sleman, Yogyakarta, Indonesia

An integrated rice–fish farming system is an efficient technique used to maintain water quality and rice/fish productivity. In addition to fish and rice plants that have mutually beneficial ecological roles, bacteria play important roles in maintaining water quality and soil fertility. Therefore, this study investigated the diversity and dynamic change in the bacterial community composition of water and sediment in an integrated rice–fish farming system using a high-throughput sequencing approach. The 16S rRNA gene of bacteria was amplified, and the IonS5™XL sequencing platform was used to identify bacterial community during the cultivation period. The results demonstrated that fishpond, rice field and trench sediment samples were dominated by the phyla Proteobacteria (60.9%–64.4%), Acidobacteria (8.7%) and Chloroflexi (6.3%), whereas water samples were dominated by the phyla Proteobacteria (65.7%), Bacteroidetes (17.1%) and Actinobacteria (8.5%). The dynamics of the bacterial community composition of the water and sediment samples were clearly observed at phylum and genus levels respectively. Water temperature (34.67°C) and ammonia-nitrogen content (0.38 mg L−1) at the early stage of cultivation correlated with the abundances of Actinobacteia and Cyanobacteria at the phylum level and Acinetobacter at the genus level. Furthermore, several bacterial genera could potentially contribute to the biogeochemical cycle. In particular, Sorangium was involved in organic matter degradation, Methylophilus was implicated in nitrification, Novosphingobium and Methylotenera were essential for denitrification, and Desulfobacca was important for sulfate reduction. This study improved our understanding of the bacterial community in the integrated rice–fish farming system and provided data for further development.

Lactobacillus reuteri BM53-1 Produces a Compound That Inhibits Sticky Glucan Synthesis by Streptococcus mutans.

Cariogenic bacteria, such as Streptococcus (S.) mutans and S. sobrinus, produce insoluble and sticky glucans as a biofilm material. The present study demonstrates that a lactic acid bacterium (LAB) named BM53-1 produces a substance that inhibits the sticky glucan synthesis. The BM53-1 strain was isolated from a flower of Actinidia polygama and identified as Lactobacillus reuteri. The substance that inhibits sticky glucan synthesis does not exhibit antibacterial activity against S. mutans. The cariogenic S. mutans produces glucans under the control of three glucosyltransferase (GTF) enzymes, named GtfB, GtfC, and GtfD. Although GtfB and GtfC produce insoluble glucans, GtfD forms soluble glucans. Through quantitative reverse-transcriptional (qRT)-PCR analysis, it was revealed that the BM53-1-derived glucan-production inhibitor (GI) enhances the transcriptions of gtfB and gtfC genes 2- to 7-fold at the early stage of cultivation. However, that of gtfD was not enhanced in the presence of the GI, indicating that the glucan stickiness produced by S. mutans was significantly weaker in the presence of the GI. Our result demonstrates that Lb. reuteri BM53-1 is useful to prevent dental caries.

Новые данные о начале функционирования Приуральского (Каргалинского) горно-металлургического центра

The article summarizes the data on the initial stage of metal production in the Southern Urals of the Bronze Age. Lots of Yamnaya culture burial mounds with copper items inside were excavated near the Kargaly deposit in the Orenburg oblast. The variety and originality of tools forms indicate the independent nature of the Ural metallurgy in the early Bronze Age. The authors present new data that allows us to reconstruct the process of metal production at the Repin (early) stage of the Yamnaya culture and explain the beginning of this process by the development of the Kargaly copper ore deposit. Excavations of the Turganik settlement were carried out. Cultural layer 5 of the early Bronze Age is dated to 3800–3360 cal BC. It is characterized by ceramics and other artefacts of the Repin type. Fragments of Kargaly copper ore, slags and copper tools (knife, awls) were found in the layer. The traceological analysis of about approximately 100 items made of stone and animal bones was performed. 41 of them are related to metallurgy and metalworking. They represent tools of all metallurgical production stages, starting from metal extraction from ore to the processing of the finished copper product. Functional groups of products such as ore mortar, ore crushing hammers, casting molds, forging hammers, anvils, edges leveling tools, sharpening stones, and others have been identified. Stone artifacts from the burials were also studied, including ore crushing and forging hammers. No mining tools were found at the settlement. It is concluded that the ore was extracted and processed at the Kargaly deposit, located 70 km to the east, and then delivered to the settlement as an enriched concentrate. The settlement was seasonal. Metallurgical activity here took its place in the summer, since the main type of economic activity was nomadic pastoralism.

Functional changes of cocultured hepatocyte sheets subjected to continuous liver regeneration stimulation in cDNA-uPA/SCID mouse: Differences in transplantation sites.

AimThe formation of a secondary liver is expected in ectopic transplants in liver therapy. It is reported that the transplantation of hepatocyte sheets constitutes one of the techniques used to form a secondary liver. Accordingly, we established a subcutaneous transplant for hepatocyte/fibroblast sheets in previous studies. In this development study with hepatocyte/fibroblast sheets, we evaluated the differences in transplantation sites to promote the maturation of transplanted tissue in a liver injury model.MethodsA cocultured hepatocyte sheet of fibroblasts (TIG-118 cells) and human hepatocytes (PXB cells) was prepared on a temperature-responsive culture dish. The prepared cocultured hepatocyte sheet was either transplanted subcutaneously or on the liver surface of a persistent liver injury model (cDNA-uPA/SCID mouse: uPA mouse), and was evaluated by the human albumin concentration in mouse blood. As a control group, hepatocyte cell sheets were used that were transplanted to both areas and compared.ResultsAlthough the cocultured hepatocyte sheet led to functional improvements in the early stages of culture in subcutaneous transplantation, these did not last in the long-term after transplantation. Although coculture effects were not observed in the liver surface transplantation case, long-term functional expressions in mono- and cocultured sheets in the case of liver surface transplantation were exhibited compared with subcutaneous administration.ConclusionThese results suggest that sustained stimulation of liver regenerationvaries depending on the transplant site and is largely involved in the maturation of hepatocyte tissue.

Integrin \u03b12\u03b21 plays an important role in the interaction between human articular cartilage-derived chondrocytes and atelocollagen gel

Although atelocollagen gel is used as a scaffold for culturing human articular cartilage-derived chondrocytes, little is known about cell–gel interactions. In this study, we investigated the mechanism via which atelocollagen gel affects human articular cartilage-derived chondrocytes. Two types of three-dimensional cultures of human articular cartilage-derived chondrocytes (i.e., with and without atelocollagen gel) were compared. While the amount of atelocollagen gel in culture gradually decreased with time, it promoted the expression of matrix metalloproteinases (MMPs) during the early stages of culture. Genome-wide differential gene expression analysis revealed that cell membrane- and extracellular matrix-related genes were highly ranked among up- and down-regulated groups in cells cultured in the presence of atelocollagen gel. Among the integrin family of genes, the expression of integrin subunit alpha 2 and integrin subunit alpha 10 was significantly increased in the presence of atelocollagen gel. Blocking α2β1 integrin with the specific inhibitor BTT 3033 had a significant effect on cell proliferation, MMP expression, and cell shape, as well as on the response to mechanical stimulation. Taken together, our findings indicate that the α2β1 integrin pathway plays an important role in the interaction of atelocollagen gel with human articular cartilage-derived chondrocytes and may be a potential therapeutic target for articular cartilage disorders.

Археолого-геофизическое исследование Зильгинского городища и Бесланского курганного катакомбного могильника в Северной Осетии

The article is devoted to new methodological techniques in the study of widely known sites of the early stage of Alanic culture of the Central Ciscaucasia — Zilgi hillfort and Beslan kurgan catacomb burial ground (RNO — Alania). The use of satellite images, low-altitude aerial photography, photogrammetry and magnetometric survey made it possible to obtain fundamentally new information about these well-studied sites and to specify their topographic and planigraphic features in the shortest time with minimal destructions. The excavations carried out in two areas of the Beslan necropolis helped to considerably refine its chronological framework, trace the development of the necropolis from the Zilgi hillfort to the southeastern periphery and identify the area of the most recent graves dating from the middle of the 7th century AD, which were made near the eastern border of unfortified settlement of Zilgi. The non-destructive survey methodology tested by the team has wide prospects for the study of Alanic culture sites of the Central Caucasus, especially at its early stage. Keywords: North Caucasus, Alanic culture, “earthen hillforts”, barrow catacomb burials, photogrammetry, Remote Sensing data, magnetometric survey