Culture Manipulation Research Articles

Effects of calcein incorporation on benthic foraminiferal community under various concentrations and incubation durations

Calcein is widely used to mark the new calcium carbonate structures in foraminifera, but it remains unclear whether calcein has effects on the biological features (i.e., abundance, size). Here the biological effects of calcein on benthic foraminifera were investigated using a two-factor crossed experiment in which foraminiferal communities (10,393 live specimens in total) were cultured in seawater spiked with calcein at four concentrations (control, 1, 5, and 10 mg/L) and for three incubation durations (2, 4, and 6 weeks). Each treatment was performed in triplicate, yielding 36 samples (4 concentrations × 3 durations × 3 replicates). Results showed that high calcein concentrations (10 mg/L) and long incubation times (6 weeks) significantly affected the abundance, and the Margalef and Shannon-Wiener diversity indices, of foraminiferal communities. Dominant species showed a species-specific response to calcein. Interestingly, different foraminiferal test types showed different responses to calcein: the proportion of miliolids significantly increased in all calcein treatments (1, 5, and 10 mg/L) relative to the control (without calcein), in contrast to the proportion of rotaliids. Thus, calcein might have a positive effect on foraminiferal communities by enhancing porcelaneous groups. Given that high calcein concentrations and long incubation times can have negative effects on foraminifera, we suggest a relatively safe manipulation: at the beginning of a culture experiment, calcein (≈5 mg/L) is added as an initial marker, then replaced with natural seawater halfway through the experiment. About 4 weeks before the end of the incubation, calcein can again be added to cultures. • 11,581 foraminiferal specimens were selected and identified as well as 10,393 living specimens analyzed. • The biological effects of calcein on benthic foraminifera were first evaluated comprehensively and analyzed quantitively. • The diverse response to calcein in different foraminiferal shells was revealed. • A relatively safe culture manipulation for foraminifera with calcein was recommended.

Timely Diagnosis of Histoplasmosis in Non-endemic Countries: A Laboratory Challenge.

Human histoplasmosis is a fungal infection caused by the inhalation of microconidia of the thermally dimorphic fungi Histoplasma capsulatum. Autochthonous cases of histoplasmosis have been diagnosed in almost every country, but it is considered an endemic infection in specific areas of the world. Many of them are popular travel destinations or the source of migratory movements. Thus, the vast majority of the registered cases in non-endemic countries are imported. They correspond to people having been exposed to the fungus in endemic locations as immigrants, expatriates, transient workers or tourists, with reported cases also associated to organ donation. Misdiagnosis and delays in initiation of treatment are not uncommon in cases of imported histoplasmosis. They are associated to high fatality-rates specially in patients with compromised cellular immunity in which progressive disseminated forms develop. The diagnosis of this infection in non-endemic countries is hampered by the lack of clinical suspicion and a dearth of available diagnostic tools adequate to offer rapid and accurate results. Non-culture-based assays such as nucleic-acid amplification tests present as a suitable alternative in this situation, offering improved sensitivity and specificity, shortened turnaround time, and increased biosafety by avoiding culture manipulation. In non-endemic regions, molecular techniques are being used mainly in laboratories from countries that have registered an increase in the incidence of imported cases. However, the number of published techniques is limited and lack consensus. Efforts are currently under way to standardize nucleic acid amplification-based techniques for its implementation in areas registering a rising number of imported cases.

A comparative in vitro study of the effect of biospecific integrin recognition processes and substrate nanostructure on stem cell 3D spheroid formation

The in vitro study of the properties of the human mesenchymal stem cells as well as their manipulation in culture for clinical purposes depends on the elimination of artefacts caused by the lack of their natural environment. It is now widely accepted that mesenchymal stem cells should be studied when they are organised as 3D spheroids rather than fibroblast-like colonies. Although this can be achieved with the use of some extracellular matrix proteins or by non-adherent conditions these suffer of significant limitations. The recent development of synthetic substrates resembling the physicochemical and biochemical properties of the adult stem cell niche has prompted questions about the role played by nanotopography and receptor-mediated adhesion. In the present paper, the influence of two types of substrates bearing the same nanostructure, but exposing either a non-specific or an integrin-specific binding motif was studied. Carboxybetaine-tethered hyperbranched poly(ɛ-lysine) dendrons showed that the hyperbranched structure was fundamental to induce spheroid formation, but these were forming more slowly, were of reduced size and less stable than those growing on substrates based on the same hyperbranched structures that had been functionalised at their uppermost branching generation by a laminin amino acid sequence, i.e. YIGSR. The study shows that both nanostructure and biorecognition need to be combined to achieve a substrate for stem cell spheroid formation as that observed in vivo in the adult stem cell niche.

Genetic Manipulation of Human Intestinal Enteroids Demonstrates the Necessity of a Functional Fucosyltransferase 2 Gene for Secretor-Dependent Human Norovirus Infection.

Human noroviruses (HuNoVs) are the leading cause of nonbacterial gastroenteritis worldwide. Histo-blood group antigen (HBGA) expression is an important susceptibility factor for HuNoV infection based on controlled human infection models and epidemiologic studies that show an association of secretor status with infection caused by several genotypes. The fucosyltransferase 2 gene (FUT2) affects HBGA expression in intestinal epithelial cells; secretors express a functional FUT2 enzyme, while nonsecretors lack this enzyme and are highly resistant to infection and gastroenteritis caused by many HuNoV strains. These epidemiologic associations are confirmed by infections in stem cell-derived human intestinal enteroid (HIE) cultures. GII.4 HuNoV does not replicate in HIE cultures derived from nonsecretor individuals, while HIEs from secretors are permissive to infection. However, whether FUT2 expression alone is critical for infection remains unproven, since routinely used secretor-positive transformed cell lines are resistant to HuNoV replication. To evaluate the role of FUT2 in HuNoV replication, we used CRISPR or overexpression to genetically manipulate FUT2 gene function to produce isogenic HIE lines with or without FUT2 expression. We show that FUT2 expression alone affects both HuNoV binding to the HIE cell surface and susceptibility to HuNoV infection. These findings indicate that initial binding to a molecule(s) glycosylated by FUT2 is critical for HuNoV infection and that the HuNoV receptor is present in nonsecretor HIEs. In addition to HuNoV studies, these isogenic HIE lines will be useful tools to study other enteric microbes where infection and/or disease outcome is associated with secretor status.IMPORTANCE Several studies have demonstrated that secretor status is associated with susceptibility to human norovirus (HuNoV) infection; however, previous reports found that FUT2 expression is not sufficient to allow infection with HuNoV in a variety of continuous laboratory cell lines. Which cellular factor(s) regulates susceptibility to HuNoV infection remains unknown. We used genetic manipulation of HIE cultures to show that secretor status determined by FUT2 gene expression is necessary and sufficient to support HuNoV replication based on analyses of isogenic lines that lack or express FUT2. Fucosylation of HBGAs is critical for initial binding and for modification of another putative receptor(s) in HIEs needed for virus uptake or uncoating and necessary for successful infection by GI.1 and several GII HuNoV strains.

Tethered primary hepatocyte spheroids on polystyrene multi-well plates for high-throughput drug safety testing

Hepatocyte spheroids are useful models for mimicking liver phenotypes in vitro because of their three-dimensionality. However, the lack of a biomaterial platform which allows the facile manipulation of spheroid cultures on a large scale severely limits their application in automated high-throughput drug safety testing. In addition, there is not yet a robust way of controlling spheroid size, homogeneity and integrity during extended culture. This work addresses these bottlenecks to the automation of hepatocyte spheroid culture by tethering 3D hepatocyte spheroids directly onto surface-modified polystyrene (PS) multi-well plates. However, polystyrene surfaces are inert toward functionalization, and this makes the uniform conjugation of bioactive ligands very challenging. Surface modification of polystyrene well plates is achieved herein using a three-step sequence, resulting in a homogeneous distribution of bioactive RGD and galactose ligands required for spheroid tethering and formation. Importantly, treatment of polystyrene tethered spheroids with vehicle and paradigm hepatotoxicant (chlorpromazine) treatment using an automated liquid handling platform shows low signal deviation, intact 3D spheroidal morphology and Z’ values above 0.5, and hence confirming their amenability to high-throughput automation. Functional analyses performance (i.e. urea and albumin production, cytochrome P450 activity and induction studies) of the polystyrene tethered spheroids reveal significant improvements over hepatocytes cultured as collagen monolayers. This is the first demonstration of automated hepatotoxicant treatment on functional 3D hepatocyte spheroids tethered directly on polystyrene multi-well plates, and will serve as an important advancement in the application of 3D tethered spheroid models to high throughput drug screening.

The pivotal role of micro-environmental cells in a human blood\u2013brain barrier in vitro model of cerebral ischemia: functional and transcriptomic analysis

BackgroundThe blood–brain barrier (BBB) is altered in several diseases of the central nervous system. For example, the breakdown of the BBB during cerebral ischemia in stroke or traumatic brain injury is a hallmark of the diseases’ progression. This functional damage is one key event which is attempted to be mimicked in in vitro models. Recent studies showed the pivotal role of micro-environmental cells such as astrocytes for this barrier damage in mouse stroke in vitro models. The aim of this study was to evaluate the role of micro-environmental cells for the functional, paracellular breakdown in a human BBB cerebral ischemia in vitro model accompanied by a transcriptional analysis.MethodsTranswell models with human brain endothelial cell line hCMEC/D3 in mono-culture or co-culture with human primary astrocytes and pericytes or rat glioma cell line C6 were subjected to oxygen/glucose deprivation (OGD). Changes of transendothelial electrical resistance (TEER) and FITC-dextran 4000 permeability were recorded as measures for paracellular tightness. In addition, qPCR and high-throughput qPCR Barrier chips were applied to investigate the changes of the mRNA expression of 38 relevant, expressed barrier targets (tight junctions, ABC-transporters) by different treatments.ResultsIn contrast to the mono-culture, the co-cultivation with human primary astrocytes/pericytes or glioma C6 cells resulted in a significantly increased paracellular permeability after 5 h OGD. This indicated the pivotal role of micro-environmental cells for BBB breakdown in the human model. Hierarchical cluster analysis of qPCR data revealed differently, but also commonly regulated clustered targets dependent on medium exchange, serum reduction, hydrocortisone addition and co-cultivations.ConclusionsThe co-cultivation with micro-environmental cells is necessary to achieve a functional breakdown of the BBB in the cerebral ischemia model within an in vivo relevant time window. Comprehensive studies by qPCR revealed that distinct expression clusters of barrier markers exist and that these are regulated by different treatments (even by growth medium change) indicating that controls for single cell culture manipulation steps are crucial to understand the observed effects properly.

Matrix stiffness regulates endosomal escape of uropathogenic E. coli.

Uropathogenic E. coli (UPEC) infection in vivo is characterized by invasion of bladder umbrella epithelial cells followed by endosomal escape and proliferation in the cytoplasm to form intracellular bacterial communities. By contrast, UPEC infection in tissue culture models results in bacteria being trapped within Lamp1-positive endosomes where proliferation is limited. Pharmacological disruption of the actin cytoskeleton has been shown to facilitate UPEC endosomal escape in vitro and extracellular matrix stiffness is a well-characterized physiological regulator of actin dynamics; therefore, we hypothesized that substrate stiffness may play a role in UPEC endosomal escape. Using functionalized polyacrylamide substrates, we found that at physiological stiffness, UPEC escaped the endosome and proliferated rapidly in the cytoplasm of bladder epithelial cells. Dissection of the cytoskeletal signaling pathway demonstrated that inhibition of the Rho GTPase RhoB or its effector PRK1 was sufficient to increase cytoplasmic bacterial growth and that RhoB protein level was significantly reduced at physiological stiffness. Our data suggest that tissue stiffness is a critical regulator of intracellular bacterial growth. Due to the ease of doing genetic and pharmacological manipulations in cell culture, this model system may provide a useful tool for performing mechanistic studies on the intracellular life cycle of uropathogens.

Long-term expansion, genomic stability and in vivo safety of adult human pancreas organoids

BackgroundPancreatic organoid systems have recently been described for the in vitro culture of pancreatic ductal cells from mouse and human. Mouse pancreatic organoids exhibit unlimited expansion potential, while previously reported human pancreas organoid (hPO) cultures do not expand efficiently long-term in a chemically defined, serum-free medium. We sought to generate a 3D culture system for long-term expansion of human pancreas ductal cells as hPOs to serve as the basis for studies of human pancreas ductal epithelium, exocrine pancreatic diseases and the development of a genomically stable replacement cell therapy for diabetes mellitus.ResultsOur chemically defined, serum-free, human pancreas organoid culture medium supports the generation and expansion of hPOs with high efficiency from both fresh and cryopreserved primary tissue. hPOs can be expanded from a single cell, enabling their genetic manipulation and generation of clonal cultures. hPOs expanded for months in vitro maintain their ductal morphology, biomarker expression and chromosomal integrity. Xenografts of hPOs survive long-term in vivo when transplanted into the pancreas of immunodeficient mice. Notably, mouse orthotopic transplants show no signs of tumorigenicity. Crucially, our medium also supports the establishment and expansion of hPOs in a chemically defined, modifiable and scalable, biomimetic hydrogel.ConclusionshPOs can be expanded long-term, from both fresh and cryopreserved human pancreas tissue in a chemically defined, serum-free medium with no detectable tumorigenicity. hPOs can be clonally expanded, genetically manipulated and are amenable to culture in a chemically defined hydrogel. hPOs therefore represent an abundant source of pancreas ductal cells that retain the characteristics of the tissue-of-origin, which opens up avenues for modelling diseases of the ductal epithelium and increasing understanding of human pancreas exocrine biology as well as for potentially producing insulin-secreting cells for the treatment of diabetes.

A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post\u2013vaccination monitoring

BackgroundVirus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post–vaccination monitoring.ResultsOne monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel cELISA was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n = 445) and C-strain VNT positive pig sera (n = 70), the 6B211 based cELISA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). The C-strain antibody can be tested in pigs as early as 7 days post vaccination with the cELISA. By testing pig sera (n = 139) in parallel, the cELISA showed excellent agreement (Kappa = 0.957) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r2 = 0.903, p < 0.001). In addition, intra- and inter-assays of the cELISA exhibited acceptable repeatability with low coefficient of variations (CVs).ConclusionsThis novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

Ideological formation: a study about the influence of contemporary music

This research aimed to investigate the influence of hard core style music of the Brazilian band DeadFish, has on the ideological formation of their followers through the internet. The methodology used was quantitative and qualitative. Data were collected online through the Google form containing seven questions designed to analyze how the lyrics of the band’s songs interfere with listeners’ habits and help them reflect on the manipulation of mass culture. As inclusion criteria, subjects who answered the questionnaire digitally between July 6 and July 7, 2019 were selected. Data collection provided 160 responses that were analyzed using the discourse analysis instrument. As a result of the analysis of the questionnaires it was possible to verify that the audience that answered the survey is not significantly affected by the lyrics of the band’s songs, to the point of making them modify consumer habits, as well as not led them to reflect more deeply. about the consumer culture imposed by the mass media. In this sense, the impact on the listeners are focused during the listening of the songs, and the message uttered by the band only lasted the time of the song, configuring itself as a form of entertainment, not surpassing this space, and not coming up with an effective criticism the band itself proposes to perform.

Cultural manipulation for the yield and quality enhancement of Pant Prabhat guava

Cultural manipulation for the yield and quality enhancement of Pant Prabhat guava

How causal are microbiomes? A comparison with the Helicobacter pylori explanation of ulcers

Human microbiome research makes causal connections between entire microbial communities and a wide array of traits that range from physiological diseases to psychological states. To evaluate these causal claims, we first examine a well-known single-microbe causal explanation: of Helicobacter pylori causing ulcers. This apparently straightforward causal explanation is not so simple, however. It does not achieve a key explanatory standard in microbiology, of Koch’s postulates, which rely on manipulations of single-microorganism cultures to infer causal relationships to disease. When Koch’s postulates are framed by an interventionist causal framework, it is clearer what the H. pylori explanation achieves and where its explanatory strengths lie. After assessing this ‘simple’, single-microbe case, we apply the interventionist framework to two key areas of microbiome research, in which obesity and mental health states are purportedly explained by microbiomes. Despite the experimental data available, interventionist criteria for explanation show that many of the causal claims generated by microbiome research are weak or misleading. We focus on the stability, specificity and proportionality of proposed microbiome causal explanations, and evaluate how effectively these dimensions of causal explanation are achieved in some promising avenues of research. We suggest some conceptual and explanatory strategies to improve how causal claims about microbiomes are made.

The Study of Emotions in Early Jewish Texts: Review and Perspectives

AbstractThis article reviews recent research on emotions in the field of early Judaism, mostly in literature. The article starts with an example from the biblical story of Joseph, to illustrate the need for a culturally sensitive understanding of emotions. Various approaches to emotions are then examined: philology and the history of the self, the construction of identity, structures of power (including gender), experiences with the divine, and emotions as adaptive practices. Each section starts with a brief outline of the scholarship conducted in other fields and serving as a background for research on early Judaism. The conclusion considers several facets of emotions, as they are highlighted by various disciplines; cultural manipulations of emotions often harness the tensions that may result from these multiple facets. The article closes with a brief assessment of the contribution of emotion research to the broader study of early Judaism and with perspectives for further research.

The Analysis of In Vivo Aging in Human Bone Marrow Mesenchymal Stromal Cells Using Colony-Forming Unit-Fibroblast Assay and the CD45lowCD271+ Phenotype

Uncultured mesenchymal stromal cells (MSCs) are increasingly used in therapies; however, the effects of donor age on their biological characteristics and gene expression remain unclear. The aim of this study was to investigate age-related changes in bone marrow (BM) MSCs following minimal or no culture manipulation. Iliac crest BM was aspirated from 67 healthy donors (19-89 years old) and directly used for the colony-forming unit-fibroblast (CFU-F) assay or CD45lowCD271+ cell enumeration. The colonies were analysed for colony area and integrated density (ID) when grown in standard MSC media or media supplemented with human serum from young (YS) or old (OS) donors. There was a notable age-related decline in the number of MSCs per millilitre of BM aspirate revealed by the CFU-F assay (r = −0.527, p < 0.0001) or flow cytometry (r = −0.307, p = 0.0116). Compared to young donors (19-40 years old), colony IDs were significantly lower in older donors (61-89 years old), particularly for smaller-sized colonies (42% lower, p < 0.01). When cultured in media supplemented with OS, young and old donor MSCs formed colonies with lower IDs, by 21%, p < 0.0001, and 27%, p < 0.05, respectively, indicating the formation of smaller sparser colonies. No significant differences in the expression of selected adipogenic, osteogenic, stromal, and bone remodelling genes as well as CD295, CD146, CD106, and connexin 43 surface molecules were found in sorted CD45lowCD271+ MSCs from young and old donors (n = 8 donors each). Altogether, these results show similar trends for age-related decline in BM MSC numbers measured by the CFU-F assay and flow cytometry and reveal age-related effects of human serum on MSC colony formation. No significant differences in selected gene expression in uncultured CD45lowCD271+ MSCs suggest that old donor MSCs may not be inferior in regard to their multipotential functions. Due to large donor-to-donor variation in all donor groups, our data indicate that an individual's chronological age is not a reliable predictor of their MSC number or potency.

Fanon’s Police Inspector

Frantz Fanon practiced psychiatry in a colonized Algeria during its struggle for independence. In his 1961 work The Wretched of the Earth, Fanon described cases from his treatment of Algerian nationalists and French colonists. I present one of Fanon’s cases as an ethical inquiry into posttraumatic stress disorder (PTSD). A French police inspector, who is employed in torture, visits Fanon with symptoms of PTSD after escalating domestic violence. The patient asks Fanon “to help him torture … with a total peace of mind.” Is it possible to treat the inspector in a meaningful way? More broadly, how might researchers and clinicians balance collective responsibilities to individual symptoms and social conditions? The answer depends on how trauma is framed: as disorder of meaning-making or circuit dysfunction, as individual illness or social rupture, as potentially gendered and racialized. These framings can reveal different views on the allocation of responsibility for the causation, expression and management of PTSD. I do not propose that it is inherently immoral to modify traumatic memories; nor do I question the efficacy of individual interventions. Rather, I ask whether PTSD has a social meaning that transcends individual comfort in decision making about erasure. What do individual interventions accomplish in the absence of concurrent political and social transformations? I argue that a holistic understanding of PTSD entails a set of social obligations: to address at its root political, gendered, and racialized violence, to repudiate occupations centered on exploitative manipulation of individuals and cultures, and to social change that prioritizes these commitments.

Anti-Fascism, Anti-Art, Doubt and Despair

Working through the artistic and theoretical contributions of anti-fascists Franco Fortini, Bertolt Brecht and Walter Benjamin, the article explores the ways in which a world lorded over by Donald Trump and his alt-right associates mobilises a Benjaminian ‘aestheticisation of politics’ alongside a populist manipulation of popular culture. Strategies developed by Brecht in 1934 to ‘restore truth’ to Nazi speeches by Goerging and Hess are used to correct the outrages of those who are shifting the atmosphere towards demagoguery, as a speech for Trump is ‘corrected’ to reveal its elitist contempt. Fortini’s poetics of doubt are also drawn on to remind the anti-fascist intellectuals of the contradictions of their positions and the dangers of dogmatism as it has been experienced historically in the communist movement. If the question is posed, through this, of the capacities of the avant-garde, it should also be noted that its attitudinal elements of pranksterism, confusionism, irony and mockery are well embedded in the fascist camp, even if the avant garde’s chief theorist, Theodore WAdorno, has become the poster boy for ‘Cultural Marxism’ and ‘snowflakes’, not least through his apparent writing of all the Beatles music. The starting question of the article is how these fragments can be accumulated and, under pressure, re-constellated to make practical sense for contemporary anti-fascist thought.

The simultaneous analysis of mesenchymal stem cells and early osteocytes accumulation in osteoarthritic femoral head sclerotic bone.

ObjectiveOA subchondral bone is a key target for therapy development. Osteocytes, the most abundant bone cell, critically regulate bone formation and resorption. Their progenitors, mesenchymal stem cells (MSCs), display altered behaviour in osteoarthritic subchondral bone. This study investigated the relationships between native osteocytes and native MSCs in osteoarthritic femoral heads.MethodsTo avoid culture manipulations, a bone treatment procedure was developed to simultaneously obtain pure osteocyte-enriched fragments and matched native CD45-CD271+ MSCs. Gene expression in osteocytes and MSCs was compared between healthy and OA bone and selected molecules were examined by immunohistochemistry in relation to OA tissue pathology. Cell sorting and standard trilineage differentiation assays were employed to test OA MSC functionality.ResultsNative osteocyte enrichment was confirmed histologically and by higher-level osteocyte maturation transcripts expression, compared with purified MSCs. Compared with healthy bone, native OA osteocytes expressed 9- and 4-fold more early/embedding osteocyte molecules E11 and MMP14, and 6-fold more osteoprotegerin (P<0.01). CD271+ MSCs accumulated in the regions of bone sclerosis (9-fold, P<0.0001) in close juxtaposition to trabeculae densely populated with morphologically immature E11-positive osteocytes (medians of 76% vs 15% in non-sclerotic areas, P<0.0001), and osteoblasts. Gene expression of OA MSCs indicated their bone formation bias, with retained multipotentiality following culture-expansion.ConclusionsIn human late-stage OA, osteogenically-committed MSCs and adjacent immature osteocytes exhibit a marked accumulation in sclerotic areas. This hitherto unappreciated MSC-early osteocyte axis could be key to understanding bone abnormalities in OA and represents a potential target for novel therapy development in early disease.

Plant tissue culture media and practices: an overview

This review presents an overview of the culture media and practices used in plant tissue culture and developmental biology. The compositions of the most commonly used basal media, especially Murashige and Skoog (MS) and modified MS (MMS), Gamborg’s B5 medium and B5 modifications, Woody Plant Medium (WPM), and Driver and Kuniyuki Woody plant medium (DKW) are discussed, along with typical basal medium manipulations to elicit and support various developmental responses. The most commonly used plant growth regulators and their applications to promote various developmental responses are examined, along with a presentation of the classical phytohormone developmental models for organogenesis and somatic embryogenesis. Elaborated developmental models for both organogenesis and somatic embryogenesis, with emphasis on discrete developmental steps, occasional need for multiple manipulations in culture to achieve a single developmental step, and identification of responsive tissue types in mixed cultures are explored. It is hoped that the information presented here will lead to a deeper understanding of basic tissue culture responses and will assist the reader in the decision-making process by identifying appropriate media and culture conditions for a particular species or application, or by providing a suitable starting point, should further customization be required.

The Wolbachia Endosymbionts.

The Wolbachia endosymbionts encompass a large group of intracellular bacteria of biomedical and veterinary relevance, closely related to Anaplasma, Ehrlichia, and Rickettsia. This genus of Gram-negative members of the Alphaproteobacteria does not infect vertebrates but is instead restricted to ecdysozoan species, including terrestrial arthropods and a family of parasitic filarial nematodes, the Onchocercidae. The Wolbachia profoundly impact not only the ecology and evolution but also the reproductive biology of their hosts, through a wide range of symbiotic interactions. Because they are essential to the survival and reproduction of their filarial nematode hosts, they represent an attractive target to fight filariasis. Their abilities to spread through insect populations and to affect vector competence through pathogen protection have made Wolbachia a staple for controlling vector-borne diseases. Estimated to be present in up to 66% of insect species, the Wolbachia are probably the most abundant endosymbionts on earth. Their success resides in their unique capacity to infect and manipulate the host germ line to favor their vertical transmission through the maternal lineage. Because the Wolbachia resist genetic manipulation and growth in axenic culture, our understanding of their biology is still in its infancy. Despite these limitations, the "-omics" revolution combined with the use of well-established and emerging experimental host models is accelerating our comprehension of the host phenotypes caused by Wolbachia, and the identification of Wolbachia effectors is ongoing.

МЕТОДОЛОГІЯ «КУЛЬТУРНОГО ПОВОРОТУ» В СУЧАСНОМУ ПЕРЕКЛАДОЗНАВСТВІ ТА ПРОБЛЕМИ АДЕКВАТНОСТІ У ПЕРЕКЛАДІ

From the above discussion it can be concluded that ‘cultural turn’ expanded the research fields of translation studies and provided new perspectives of translation. It demonstrated accent on native culture and flexible translation. The accent was shifted from the text to the reader’s reception and the quality of translation depended on it. At the same time, consideration of native culture, social background and tradition led to natural literature for native readers. Besides, translation was regarded as a method of manipulation and creation of certain culture.