Specific Culture Conditions Research Articles

Efficient derivation of embryonic stem cells and primordial germ cell-like cells in cattle.

The induction of the germ cell lineage from pluripotent stem cells (in vitro gametogenesis) will help understand the mechanisms underlying germ cell differentiation and provide an alternative source of gametes for reproduction. This technology is especially important for cattle, which are among the most important livestock species for milk and meat production. Here, we developed a new method for robust induction of primordial germ cell-like cells (PGCLCs) from newly established bovine embryonic stem (bES) cells. First, we refined the pluripotent culture conditions for pre-implantation embryos and ES cells. Inhibition of RHO increased the number of epiblast cells in the pre-implantation embryos and dramatically improved the efficiency of ES cell establishment. We then determined suitable culture conditions for PGCLC differentiation using bES cells harboring BLIMP1-tdTomato and TFAP2C-mNeonGreen (BTTN) reporter constructs. After a 24-h culture with bone morphogenetic protein 4 (BMP4), followed by three-dimensional culture with BMP4 and a chemical agonist and WNT signaling chemical antagonist, bES cells became positive for the reporters. A set of primordial germ cells (PGC) marker genes, including PRDM1/BLIMP1, TFAP2C, SOX17, and NANOS3, were expressed in BTTN-positive cells. These bovine PGCLCs (bPGCLCs) were isolated as KIT/CD117-positive and CD44-negative cell populations. We anticipate that this method for the efficient establishment of bES cells and induction of PGCLCs will be useful for stem cell-based reproductive technologies in cattle.

Творчество Саят-Новы в свете интерпретации и перевода В. Брюсова

Each national literature is a part of national history. It presents the features of the national culture and the people who live and develop in specific cultural and traditional conditions. Each nation has its unique genius that sings the national beauty. Armenian ashug Sayat’-Nova’s creativity has had a significant influence on Armenian literature. Sayat’-Nova, as a national poet, had created poetry with its genre subdivisions. We can consider such categories as spiritual, patriotic poetry, glamorous social poetry, and folk-gusan songs. The present research has been carried out along the lines of Sayat’-Nova’s verbal creativity through the interpretive prism of theorists and literary critics with special reference to Valeri Brusov, who highlighted the exclusive richness of Sayat’-Nova’s artistic devices, the peculiar features of his literary genre, style, and language. The article focused on the relevance of Russian-Armenian literary relations, which became particularly crucial with the publication of Valeri Brusov’s anthology “Poetry of Armenia” in 1916. The outstanding Russian writer, critic, and translator has highly appreciated the exceptional richness of Armenian literature and interpreted each piece of verbal creativity with great accuracy. His opinion is that Sayat’-Nova name deserves to be a part of world literature. Armenian ashug has to stand next to the honored names and glorified works of Western literature.

Succession and Cultural Interaction of the Worldview of the Kazakh People with Ancient Greek Mythology and Philosophy

The main idea of the work is the concept of the formation of philosophy in Ancient India and Ancient Greece, based on the initial close connection of myth, religion, and science, which subsequently contributes to the separation of theoretical philosophy as a form of knowledge based on logic and pure reason. In the article, based on a summary of scientific studies published in recent years, it is shown that this trend is characteristic of the Turkic people, including the Kazakh people, who also did not stay away from this process. Each person, thanks to oral literature and folklore, which accumulates the basis of popular worldview, indulges in fleeting fantasies about life, and develops the ability to independently think about the existence of the world. The article also analyzes the question of the influence of the nomadic way of life on the perfection of deep thinking of the steppe people. The author comes to the conclusion that, despite the specific historical and cultural conditions, the knowledge and ideas of the ancient Greeks and the Kazakh people about the world, being, man, society, nature and life are very closely intertwined and demonstrate a deep connection. Such well-known scientists as Auezkhan Kodar, Seit Kaskabasov, Sagadi Bulekbaev in their studies of biogenetic laws and altered states of consciousness, based on typological analysis, note that this continuity and interrelationship of worldviews is not accidental, and cultural dialogue is not limited to a certain historical period. This is a continuous process that is reflected in the consciousness of each new generation, and thus goes beyond the space-time continuum. In their mythological knowledge, both peoples were among the first to try to generalize, classify and understand the systematicity, diversity and multiplicity of the universe. The result is the conclusion that both the formation of knowledge about the surrounding world and the realization of oneself and the meaning of life took place through the synthesis of conceptual thinking and artistic universals

Politico-Cultural Exigencies and the Rise of Rhetoric in Classical Greece

Did rhetoric naturally evolve from the inherent power and potential of language, or were specific political and cultural conditions instrumental in driving its development in classical Greece? This paper explores this question by adopting a historical approach to rhetoric and a rhetorical approach to history. It draws on the perspectives of sophists such as Gorgias, the anonymous author of “Dissoi Logoi,” and Isocrates, as well as the rhetorical philosophical orientations of Plato and Aristotle. In conducting this study, I employ a methodological review approach that relies on the scholarship of rhetoric and rhetorical history and critically analyse available sources and the arguments developed in the field. Through this approach, I argue that the emergence of rhetoric in classical times was prompted by several exigent factors, including humanism, democracy, education, literacy, and legal practices. Therefore, theorizing and practicing rhetoric beyond the cultural-political context is inherently partial and incomplete.

Assessment of Fallopian Tube Epithelium Features Derived from Induced Pluripotent Stem Cells of Both Fallopian Tube and Skin Origins.

Fallopian tube epithelial cells (FTECs) play a significant role in the development of high-grade serous ovarian cancer (HGSOC), but their utilization in in vitro experiments presents challenges. To address these limitations, induced pluripotent stem cells (iPSCs) have been employed as a potential solution, driven by the hypothesis that orthologous iPSCs may offer superior differentiation capabilities compared with their non-orthologous counterparts. Our objective was to generate iPSCs from FTECs, referred to as FTEC-iPSCs, and compare their differentiation potential with iPSCs derived from skin keratinocytes (NHEK). By introducing a four-factor Sendai virus transduction system, we successfully derived iPSCs from FTECs. To assess the differentiation capacity of iPSCs, we utilized embryoid body formation, revealing positive immunohistochemical staining for markers representing the three germ layers. In vivo tumorigenesis evaluation further validated the pluripotency of iPSCs, as evidenced by the formation of tumors in immunodeficient mice, with histological analysis confirming the presence of tissues from all three germ layers. Quantitative polymerase chain reaction (qPCR) analysis illuminated a sequential shift in gene expression, encompassing pluripotent, mesodermal, and intermediate mesoderm-related genes, during the iPSC differentiation process into FTECs. Notably, the introduction of WNT3A following intermediate mesoderm differentiation steered the cells toward a FTEC phenotype, supported by the expression of FTEC-related markers and the formation of tubule-like structures. In specific culture conditions, the expression of FTEC-related genes was comparable in FTECs derived from FTEC-iPSCs compared with those derived from NHEK-iPSCs. To conclude, our study successfully generated iPSCs from FTECs, demonstrating their capacity for FTEC differentiation. Furthermore, iPSCs originating from orthologous cell sources exhibited comparable differentiation capabilities. These findings hold promise for using iPSCs in modeling and investigating diseases associated with these specific cell types.

Formidling av det fragmenterte

The development of musical drama in Norway took place during specific cultural and economic conditions which contributed both to a sporadic performance tradition and a number of unfinished or unperformed works. The history of musical drama in Norway, then, is partly a history of that which did not take place.
 The field’s archival situation remains partly opaque, with music scores, libretti, and other source materials dispersed across libraries, archives and museums; in private ownership or presumed lost. This has had historiographic consequences both for the perspective of theatre and music history studies, as well as for the performing arts.
 The recently founded Society for Norwegian Musical Drama Heritage (FNMA) has initiated the creation of a digital compendium to increase the accessibility of established knowledge as well as of the historical source materials of Norwegian musical drama from the mid-1750s up to the 2nd World War.
 Taking as their starting point an understanding of the dispersed history of musical drama – performed and unperformed – as an unarranged archive, or reservoir, the authors discuss the potential of a digital compendium as a cultural heritage tool.

Integrated-Omics Study on the Transcriptomic and Metabolic Changes of Bacillus licheniformis, a Main Microorganism of Fermented Soybeans, According to Alkaline pH and Osmotic Stress.

Bacillus licheniformis has been widely utilized in the food industry as well as various agricultural industries. In particular, it is a main microorganism of fermented soybeans. In this study, the changes of the metabolome and transcriptome of B. licheniformis KACC15844, which had been isolated from fermented soybeans, were investigated depending on alkaline pH (BP) and a high salt concentration (BS) using an integrated-omics technology, focusing on leucine metabolism. Overall, carbohydrate (glycolysis, sugar transport, and overflow) and amino acid (proline, glycine betaine, and serine) metabolisms were strongly associated with BS, while fatty acid metabolism, malate utilization, and branched-chain amino acid-derived volatiles were closely related to BP, in both gene and metabolic expressions. In particular, in leucine metabolism, the formation of 3-methylbutanoic acid, which has strong cheesy odor notes, was markedly increased in BP compared to the other samples. This study provided information on how specific culture conditions can affect gene expressions and metabolite formations in B. licheniformis using an integrated-omics approach.

Induction of Fungal Secondary Metabolites by Co-Culture with Actinomycete Producing HDAC Inhibitor Trichostatins.

A recently bioinformatic analysis of genomic sequences of fungi indicated that fungi are able to produce more secondary metabolites than expected. Despite their potency, many biosynthetic pathways are silent in the absence of specific culture conditions or chemical cues. To access cryptic metabolism, 108 fungal strains isolated from various sites were cultured with or without Streptomyces sp. 13F051 which mainly produces trichostatin analogues, followed by comparison of metabolic profiles using LC-MS. Among the 108 fungal strains, 14 produced secondary metabolites that were not recognized or were scarcely produced in mono-cultivation. Of these two fungal strains, Myrmecridium schulzeri 15F098 and Scleroconidioma sphagnicola 15S058 produced four new compounds (1-4) along with a known compound (5), demonstrating that all four compounds were produced by physical interaction with Streptomyces sp. 13F051. Bioactivity evaluation indicated that compounds 3-5 impede migration of MDA-MB-231 breast cancer cells.

Fever of Unknown Origin in a Patient Administered With Bacillus Calmette-Guérin (BCG) to Treat Urothelial Vesical Carcinoma in Situ

Koch’s Bacillus bovis (Mycobacterium bovis) was made avirulent by special culture conditions, i.e., 230 passages on medium containing potatoes treated with bile salts. It has been used mainly to prevent tuberculosis. The vaccine was named after the bacteriologist Albert Calmette and the veterinarian Camille Guérin, i.e., Bacillus Calmette-Guérin (BCG). These researchers worked at the Pasteur Institute in Lille and launched the vaccine in 1921.Recently, BCG was used also for its non-specific immunostimulant action. Intravesical administration of BCG is an adjunctive therapy for the treatment of bladder cancer, which does not invade the muscle wall.We will discuss the clinical case of a patient who had fever of unknown origin for about 2 months and was resistant to treatment with multiple lines of antibiotics.

SecretoMyc, a web-based database on mycobacteria secreted proteins and structure-based homology identification using bio-informatics tools

To better understand the interaction between the host and the Mycobacterium tuberculosis pathogen, it is critical to identify its potential secreted proteins. While various experimental methods have been successful in identifying proteins under specific culture conditions, they have not provided a comprehensive characterisation of the secreted proteome. We utilized a combination of bioinformatics servers and in-house software to identify all potentially secreted proteins from six mycobacterial genomes through the three secretion systems: SEC, TAT, and T7SS. The results are presented in a database that can be crossed referenced to selected proteomics and transcriptomics studies (https://secretomyc.cbs.cnrs.fr). In addition, thanks to the recent availability of Alphafold models, we developed a tool in order to identify the structural homologues among the mycobacterial genomes.

Getting More Women on Boards: Cultural and Institutional Antecedents That Matter

The purpose of this study is to contribute to a better understanding of the multiple cultural and institutional antecedents which can positively (or negatively) impact the incorporation of more women on corporate boards (WoCB). Many contemporary discourses about quotas versus voluntary actions by corporations as drivers of greater gender diversity on boards are largely based on cross-country mimicking. These discourses often fail to integrate country-level configurations and conditions, as drivers or barriers to change. We advance this research by seeking to understand the complexity of such multiple antecedents to getting WoCB. Results show multiple macro factors involved in getting WoCB in Europe. We test the complex inter-relations of factors such as legislation, cultural, societal, economic, political, and institutional variables, and their effects. Findings highlight important drivers of more women on boards such as gender equality in political empowerment and institutional settings; government-regulated maternity and paternity leave; specific legislative or soft quota policies; and societal cultural variables, such as (low) power distance and (high) assertiveness, as drivers of getting WoCB. These results prove the need to consider and complement legislative policies to specific cultural and institutional conditions in each country. To help policymakers, we provide insights into which multiple macro factors act as drivers or barriers in their organizations or societies for getting more WoCB. This will help decision makers in organizations or policymaking bodies to match gender diversity goals to the multiple country-level conditions they need to navigate, hence making a better world together.

P-198 Evolution of artificial intelligence-based embryo selection models: a massive external validation on 70,456 embryos

Abstract Study question How do three artificial intelligence-based embryo selection models that have evolved over the years work? Summary answer The three automatic scores were positively associated with clinical outcomes, and the iDAScore v2 showed the best performance in conventional treatments with patient oocytes. What is known already Artificial intelligence (AI) models have been introduced in in vitro fertilization laboratories in recent years as an adjunct to clinical decision-making. The most initial function of the AI involved guiding embryologists in the annotations of embryonic events. This was followed by the development of embryo selection models based on machine learning, requiring annotations (i.e., KIDScore). Finally, the latest in this field are models based on deep learning, which analyze raw time-lapse video (i.e., iDAScore). In this study, the evolution of three AI-based models have been analyzed on the same large set of embryos. Study design, size, duration This single-center study includes 6,737 patients who underwent in vitro fertilization treatments for 6 consecutive years. A total of 7,722 cycles were analyzed, resulting in 70,456 embryos cultured in EmbryoScope® time-lapse systems. Embryos were routinely evaluated and selected according to conventional morphology (ASEBIR criteria) by senior embryologists. Retrospectively, the embryos were scored by three AI models from 1 to 9,9: KIDScore D5 v3 (n = 32,784), iDAScore v1 (n = 68,440) and iDAScore v2 (n = 68,471). Participants/materials, setting, methods Automatic embryo scores were compared with conventional morphology, ploidy, and clinical outcomes for single blastocyst transfers. Then, we performed multivariate logistic regression analysis (confounding factors: oocyte origin, donated-autologous; type of embryo transfer, fresh-frozen; oocyte age; patient body mass index; culture strategy, individual-group; day of embryo transfer, fifth-sixth day of embryo development) in different patient populations (PGT-A, oocyte donation program and conventional treatments with patient oocytes). Finally, the performance (AUC) was calculated for comparison. Main results and the role of chance The mean of the three scores increased as embryos had better morphological grade*. Regarding ploidy (euploid vs. aneuploid): 5,62 ± 1,78 vs. 5,00 ± 1,72 for KIDScore* (n = 6,580); 7.59 ± 1.61 vs. 6.92 ± 1.75 for iDAScorev1* (n = 7,089); and 6.31 ± 2.53 vs. 5.07 ± 2.55 for iDAScorev2* (n = 7,082). Regarding implantation (implanted vs. non-implanted): 6.24 ± 2.01 vs. 5.42 ± 2 for KIDScore* (n = 9,681); 8.36 ± 1.37 vs. 7.76 ± 1.70 for iDAScorev1* (n = 10,079); and 6.83 ± 2.18 vs. 5.75 ± 2.54 for iDAScorev2* (n = 10,068). Regarding live birth (positive vs. negative): 6.29 ± 2.01 vs. 5.51 ± 2.01 for KIDScore* (n = 9,668); 8,39 ± 1,34 vs.7,83 ± 1,68 for iDAScorev1* (n = 10,065); and 6.88 ± 2.15 vs. 5.87 ± 2.52 for iDAScorev2* (n = 10,054). In general, the multivariate analysis showed statistically significant odds ratio for the three models in predicting implantation and live birth (all patient subpopulations)*. Regarding oocyte donation program: the AUCs for predicting implantation were 0.636 [0.621−0.650] for KIDScore, 0.638 [0.624−0.652] for iDAScorev1, and 0.636 [0.622−0.650] for iDAScorev2; and the AUCs for live birth prediction were 0.630 [0.615−0.644] for KIDScore, 0.629 [0.615−0.643] for iDAScorev1, and 0.635 [0.621−0.649] for iDAScorev2. Regarding treatments with patient oocytes: the AUCs for predicting implantation were 0.667 [0.644−0.689] for KIDScore, 0.674 [0.652−0.696] for iDAScorev1, and 0.686 [0.664−0.708] for iDAScorev2; and the AUCs for live birth prediction were 0.664 [0.642−0.687] for KIDScore, 0.669 [0.647−0.692] for iDAScorev1, and 0.686 [0.664−0.708] for iDAScorev2. *p<0.001 Limitations, reasons for caution A major limitation of our study is its retrospective nature. Although ours is the largest external validation ever performed with an unselected ICSI population, its single-center design should be considered for the universal application of the models. Also, specific culture conditions should be addressed when considering the generalized application. Wider implications of the findings Our results showed a positive association between automatic scores and the success of IVF. Despite observing similar AUCs between the three embryo selection models, the use of the most advanced automatic algorithms should improve workflow, standardize the process between laboratories and allow embryologists to spend their time on other tasks. Trial registration number PI21/00283

P-174 Do culture conditions alter the efficacy of embryo selection algorithms using time-lapse technology? Development of novel embryo selection model with embryos cultured in different conditions

Abstract Study question Is the efficacy of embryo selection models altered by the conditions employed for embryo culture? Summary answer The predictive capacity of morphology and morphokinetics-based selection models might be dependent on the incubator and culture conditions employed. What is known already Morphological assessment remains the gold-standard for embryo selection. However, since the introduction of time-lapse technology to the incubators used for embryo culture, many selection algorithms based on morphokinetic annotations have been developed, adding objectivity to the embryo selection process. The efficacy and reproducibility of these algorithms have been questioned by other authors, being potentially influenced by characteristics of the patients and differing culture conditions. Nowadays, there are several incubators with time-lapse technology in the market, some of them providing novel features such as culture in high humidity conditions, which has been related to differences in the morphokinetics of embryo development. Study design, size, duration Retrospective external validation of an embryo selection algorithm based on morphokinetic annotations, in a set of 555 transferred blastocysts cultured in a time-lapse system in dry (DC, n = 281) or humid conditions (HC, n = 274), and comparison with selection by morphological criteria. A novel selection model was developed considering morphokinetic annotations of our embryo dataset, including blastocysts cultured in DC and HC. Embryos belong to autologous and oocyte-donation ICSI cycles performed in a clinic over 3 years. Participants/materials, setting, methods Embryos were cultured in a Geri incubator (Genea Biomedx) and automatically annotated (Connect&Assess2.0). The efficacy of the algorithm published by Motato et al, 2016, and ASEBIR morphological grading was assessed by Generalized Estimating Equations (GEE), considering possible confounders. Efficacy was quantified by the Area Under the ROC Curve (AUC), its 95% confidence interval (CI) and statistical significance was assessed by the Mann-Whitney test. A novel algorithm was developed aided by the visual tool FertAI (Merck). Main results and the role of chance Transferred blastocysts with known implantation data were classified A-D in base of the algorithm published by Motato et al, 2016, according to the optimal range of 2 morphokinetic parameters, tEB and s3, empirically-obtained in embryos cultured in an Embryoscope incubator. The algorithm had an AUC = 0.591, 95%CI(0.542−0.64), resulting significantly predictive of implantation (P<0.001), but lower than the efficacy reported by the authors (AUC = 0.602, 95%CI(0.559−0.645)). The efficacy was different in the two culture conditions: AUC(DC) = 0.608, 95%CI(0.54−0.676), P = 0.002; AUC(HC) = 0.588, 95%CI(0.518−0.657), P = 0.103. The morphological evaluation (3 categories: A (best) to C (worse)) resulted statistically predictive of implantation: AUC of 0.596, 95%CI(0.547−0.646), P<0.001. Again, its efficacy was different in DC (AUC = 0.626, 95%CI(0.559−0.693), P<0.001) and HC (AUC = 0.589, 95%CI(0.52−0.657), P = 0.013). The lower efficacy shown by these algorithms might be associated with different morphokinetic development of embryos cultured in a different incubator and different culture conditions. Hence, a novel scoring model was developed with empirically-determinated optimal ranges of three morphokinetic parameters, considering, for the first time, embryos cultured in DC and HC: tEB<113.874; (t5-t3)/(t5-t2) = [0.521, 0.554] and cc2 = [10.34, 11.58], yielding a score from 0 to 3. The selection model resulted in an AUC = 0.637, 95%CI(0.59−0.684), and was equally efficient in DC and HC: AUC(DC) = 0.645, 95%CI(0.579−0.712); AUC(HC) = 0.645, 95%CI(0.579−0.711); P<0.001). Limitations, reasons for caution This is a primary approach to a development of a selection algorithm using morphokinetic data of embryos cultured until blastocyst stage in a Geri incubator. The efficacy and reproducibility of the model must be validated in a different dataset. Wider implications of the findings The lower efficacy shown by a selection algorithm developed in a different incubator supports the necessity of adjusting selection tools for the specific culture conditions employed by each IVF laboratory. This is the first scoring model developed for selection of blastocysts using morphokinetic parameters recorded in a Geri time-lapse incubator. Trial registration number Not applicable

Blastospores from Metarhizium anisopliae and Metarhizium rileyi Are Not Always as Virulent as Conidia Are towards Spodoptera frugiperda Caterpillars and Use Different Infection Mechanisms.

Infective conidia from entomopathogenic fungi are widely used to control insect pests. Many entomopathogenic fungi also produce yeast-like cells called blastospores under specific liquid culture conditions that can directly infect insects. However, little is known about the biological and genetic factors that allow blastospores to infect insects and make them potentially effective for biological control in the field. Here, we show that while the generalist Metarhizium anisopliae produces a higher number of and smaller blastospores, the Lepidoptera specialist M. rileyi produces fewer propagules with a higher cell volume under high-osmolarity conditions. We compared the virulence of blastospores and conidia of these two Metarhizium species towards the economically important caterpillar pest Spodoptera frugiperda. Conidia and blastospores from M. anisopliae were equally infectious, but acted slower, and killed fewer insects than M. rileyi conidia and blastospores did, where M. rielyi conidia had the highest virulence. Using comparative transcriptomics during propagule penetration of insect cuticles, we show that M. rileyi blastospores express more virulence-related genes towards S. frugiperda than do M. anisopliae blastospores. In contrast, conidia of both fungi express more virulence-related oxidative stress factors than blastospores. Our results highlight that blastospores use a different virulence mechanism than conidia use, which may be explored in new biological control strategies.

BIOL-11. PRECLINICAL MODELLING OF PEDIATRIC BRAIN TUMORS USING ORGANOID TECHNOLOGY

Abstract Molecular characterization has resulted in improved classification of pediatric brain tumors, leading to many novel (sub)types with distinct oncodriving events. To study tumor biology and to perform translational research on each of these tumors, preclinical models are essential. However, we are currently lacking sufficient models, especially in vitro, to represent each (sub)type and their heterogeneity. To generate large series of preclinical in vitro models for pediatric brain tumors, we are using organoid technology. Cells from patient samples and patient-derived xenograft samples have been taken into culture to establish 3D organoids using tumor type specific culture conditions. These organoid lines retain the molecular characteristics of the original tumor tissue. They can be used to perform high-throughput drug screens, genetic manipulations, and co-cultures with, for instance, immune cells. Viable tissue is not always available for all tumor (sub)types and specific oncodrivers. To circumvent this lack of tissue, we can also induce tumors in vitro. Therefore, we generate cerebral and cerebellar brain organoids from human pluripotent stem cells. These organoids mimic human developing brain cells and can be genetically manipulated to model different brain tumor types. These genetically engineered brain tumor models allow us to study the cellular origins of pediatric brain tumors and the different tumor driving mechanisms. Tumors induced in the brain organoids histologically and molecularly resemble human patient samples based on (single cell) transcriptomic analyses. Moreover, the tumor cells are able to establish xenografts in mouse brains. In summary, organoid technology provides a novel avenue to establish in vitro models for pediatric brain tumors. At the meeting we will present data for various new ependymoma, medulloblastoma and embryonal brain tumor organoid models.

A concise in vitro model for evaluating interactions between macrophage and skeletal muscle cells during muscle regeneration.

Skeletal muscle has a highly regenerative capacity, but the detailed process is not fully understood. Several in vitro skeletal muscle regeneration models have been developed to elucidate this, all of which rely on specialized culture conditions that limit the accessibility and their application to many general experiments. Here, we established a concise in vitro skeletal muscle regeneration model using mouse primary cells. This model allows evaluation of skeletal muscle regeneration in two-dimensional culture system similar to a typical cell culture, showing a macrophage-dependent regenerative capacity, which is an important process in skeletal muscle regeneration. Based on the concept that this model could assess the contribution of macrophages of various phenotypes to skeletal muscle regeneration, we evaluated the effect of endotoxin pre-stimulation for inducing various changes in gene expression on macrophages and found that the contribution to skeletal muscle regeneration was significantly reduced. The gene expression patterns differed from those of naive macrophages, especially immediately after skeletal muscle injury, suggesting that the difference in responsiveness contributed to the difference in regenerative efficiency. Our findings provide a concise in vitro model that enables the evaluation of the contribution of individual cell types, such as macrophages and muscle stem cells, on skeletal muscle regeneration.

Potential bio-herbicide from Streptomyces sp. KR0005 to control weeds in a horticultural field

ABSTRACT Herbicidal active metabolites obtained from actinomycetes provide valuable sources in the search for new candidates. They might be striking alternatives to synthetic herbicides. In the present study, actinobacteria strains were isolated from soil samples and their herbicidal activities were screened. In the screening, Streptomyces sp. strain KR0005 broth culture exhibited the highest phytotoxic activity. Thus, it was selected for further studies. The herbicidal active compound purified from this selected strain was identified as cycloheximide (MW, 281.; C15H23NO4) based on 1H-NMR, 13C-NMR, 2D-NMR, and high-resolution electrospray ionisation mass spectrometry analyses. Dose–response experiments revealed that the GR50 value of Digitaria ciliaris was 1.37 g L−1 to Streptomyces sp. strain KR0005 culture filtrate. In general, the inclusion of v/v 0.1% of LES270 (Alcohols, (C12-14), ethoxylated, mono ethers with sulphuric acid, sodium salts) and SF90 (Alcohols, C12-14-secondary, ethoxylated) surfactants resulted in a 14.3% decrease and a 24.1% increase in herbicidal activity, respectively, compared with Tween-20 (Polyoxyethylene sorbitan monolaurate). Mode of action experiments revealed that the KR0005 culture filtrate acted as both photosynthesis and cell membrane integrity inhibitors in a dose-dependent manner. However, further studies such as mutation and/or specific culture conditions are needed to increase the herbicidal activity of this selected strain.

Ocular Mycobacterium haemophilum infection originating in the cornea: a case report

BackgroundMycobacterium haemophilum is a slow-growing non-chromogenic nontuberculous Mycobacterium species that can cause skin infection or arthritis in an immunocompromised population or in children. Primary infection of the healthy adult cornea is rare. The special requirements for culture make this pathogen difficult to diagnose. The study aims to report the clinical manifestation and treatment process of corneal infection and notify the awareness of M. Haemophilus keratitis among clinicians. This is the first case report of primary M. haemophilum infection in the cornea of healthy adults reported in the literature.Case presentationA 53-year-old healthy goldminer presented with left eye redness and a history of vision loss for four months. The patient was misdiagnosed with herpes simplex keratitis until M. haemophilum was detected using high-throughput sequencing. Penetrating keratoplasty was performed, and a large number of mycobacteria were detected by Ziehl-Neelsen staining of the infected tissue. Three months later, the patient developed conjunctival and eyelid skin infections that manifested as caseous necrosis of the conjunctiva and skin nodules. After excision and debridement of the conjunctival lesions and systemic antituberculosis drug treatment for 10 months, the patient was cured.ConclusionM. haemophilum could cause primary corneal infection in healthy adults, which is an infrequent or rare infection. Owing to the need for special bacterial culture conditions, conventional culture methods do not provide positive results. High-throughput sequencing can rapidly identify the presence of bacteria, which aids in early diagnosis and timely treatment. Prompt surgical intervention is an effective treatment option for severe keratitis. Long-term systemic antimicrobial therapy is crucial.

Analyzing Mitochondrial Morphology Through Simulation Supervised Learning

The quantitative analysis of subcellular organelles such as mitochondria in cell fluorescence microscopy images is a demanding task because of the inherent challenges in the segmentation of these small and morphologically diverse structures. In this article, we demonstrate the use of a machine learning-aided segmentation and analysis pipeline for the quantification of mitochondrial morphology in fluorescence microscopy images of fixed cells. The deep learning-based segmentation tool is trained on simulated images and eliminates the requirement for ground truth annotations for supervised deep learning. We demonstrate the utility of this tool on fluorescence microscopy images of fixed cardiomyoblasts with a stable expression of fluorescent mitochondria markers and employ specific cell culture conditions to induce changes in the mitochondrial morphology.

Analyzing Mitochondrial Morphology Through Simulation Supervised Learning

The quantitative analysis of subcellular organelles such as mitochondria in cell fluorescence microscopy images is a demanding task because of the inherent challenges in the segmentation of these small and morphologically diverse structures. In this article, we demonstrate the use of a machine learning-aided segmentation and analysis pipeline for the quantification of mitochondrial morphology in fluorescence microscopy images of fixed cells. The deep learning-based segmentation tool is trained on simulated images and eliminates the requirement for ground truth annotations for supervised deep learning. We demonstrate the utility of this tool on fluorescence microscopy images of fixed cardiomyoblasts with a stable expression of fluorescent mitochondria markers and employ specific cell culture conditions to induce changes in the mitochondrial morphology.