Culture Methods Research Articles

Potential Use of Organoids in Regenerative Medicine.

In vitro cell culture is crucial for studying human diseases and development. Compared to traditional monolayer cultures, 3D culturing with organoids offers significant advantages by more accurately replicating natural tissues' structural and functional features. This advancement enhances disease modeling, drug testing, and regenerative medicine applications. Organoids, derived from stem cells, mimic tissue physiology in a more relevant manner. Despite their promise, the clinical use of regenerative medicine currently needs to be improved by reproducibility, scalability, and maturation issues. This article overviews recent organoid research, focusing on their types, sources, 3D culturing methods, and applications in regenerative medicine. A literature review of "organoid" and "regenerative medicine" in PubMed/MEDLINE highlighted relevant studies published over the past decade, emphasizing human-sourced organoids and their regenerative benefits, as well as the availability of free full-text articles. The review uses descriptive data, including tables and text, to illustrate the challenges and potential of organoids in regenerative medicine. The transition from 2D to 3D models, particularly organoids, has significantly advanced in vitro research. This review covers a decade of progress in various organoid types-such as liver, cholangiocyte, intestinal, pancreatic, cardiac, brain, thymus, and mammary organoids-and their 3D culture methods and applications. It addresses critical issues of maturity, scalability, and reproducibility and underscores the need for standardization and improved production techniques to facilitate broader clinical applications in regenerative medicine. Successful therapy requires increased scalability and standardization. Organoids have enormous potential in biological research, notwithstanding obstacles.

Comparison of qPCR and chromogenic culture methods for rapid detection of group B streptococcus colonization in Vietnamese pregnant women

IntroductionNeonatal infections can rapidly become severe, with delays in treatment often proving fatal. Streptococcus agalactiae (Group B Streptococcus, GBS) is a common cause, typically transmitted from colonized pregnant women to neonates during childbirth. In Vietnam, routine prenatal care lacks standardized GBS screening protocols. This study aims to compare enhanced qPCR methods with the culture method, evaluate the diagnostic accuracy of these qPCR procedures, and assess the frequency of GBS infection in pregnant Vietnamese women during their final trimester. Materials and methodsPregnant women aged 35 weeks gestation or more were recruited. Rectovaginal swabs were collected and analyzed for GBS using chromogenic culture, a commercial real-time PCR kit, and in-house real-time PCR assays targeting the cfb and sip genes. Clinical diagnostic values were calculated, and GBS prevalence was determined. ResultsThe study included 259 pregnant women with a mean age of 30.2 ± 5.0 years. Of these, 96.6 % had gestational ages of 37 weeks or more at delivery. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the cfb-based and sip-based qPCR assays were 94.1/92.7, 99.0/99.5, 97.1/98.5, 97.8/97.3, and 97.6 %, respectively. The Kappa values were excellent (0.940 and 0.939), with results available in under 2 h. GBS prevalence was 24.7 % and 25.5 % by cfb-based and sip-based qPCR assays, aligning with the culture method (25.5 %). ConclusionsBoth direct real-time PCR assays demonstrated high accuracy and were comparable to chromogenic culture in diagnosing GBS. A significant prevalence of GBS colonization was found among Vietnamese pregnant women in their final trimester.

Landowner economy in the Kazan province in the first half of the 19th century

The article is devoted to the economic development of landowners in the Kazan province during the era of serfdom. The source base of the study contains published memoirs of representatives of the nobility, statistical publications, pre-revolutionary studies of the state of agriculture in private farms, as well as archival material containing economic descriptions of 12 districts of the province, compiled by local leaders of the nobility and individual landowners based on the questions of the questionnaire of the Free Economic Society in 1802. The author analyzed the natural and geographical factors that influenced economic life on the estates; characterized the working inventory and showed attempts to modernize it; studied the specialization of estates and identified the economy-forming industries; considered the features of the corvee and quitrent systems of peasant exploitation. In addition, the article shows examples of organizing market-oriented agricultural production. A conclusion is made about the subsistence nature of the landowner economy in the Kazan province in the pre-reform period, in which the sale of surplus did not mean the commercial nature of production. The availability of free labor deprived the incentive to introduce improved methods of land cultivation – use modern equipment, improved varieties of crops, as well as develop industries such as livestock and crop production. For citation: Mironova E.V. Landowner economy in the Kazan province in the first half of the 19th century. From History and Culture of Peoples of the Middle Volga Region. 2024, vol.14, no.3, pp.118–136. https://doi.org/10.22378/2410-0765.2024-14-3.118-136 (In Russian)

Selective colonization of microplastics, wood and glass by antimicrobial-resistant and pathogenic bacteria.

The Plastisphere is a novel niche whereby microbial communities attach to plastic debris, including microplastics. These communities can be distinct from those found in the surrounding environment or those attached to natural substrates and may serve as a reservoir of both pathogenic and antimicrobial-resistant (AMR) bacteria. Owing to the frequent omission of appropriate comparator particles (e.g. natural substrates) in previous studies, there is a lack of empirical evidence supporting the unique risks posed by microplastics in terms of enrichment and spread of AMR pathogens. This study investigated selective colonization by a sewage community on environmentally sampled microplastics with three different polymers, sources and morphologies, alongside natural substrate (wood), inert substrate (glass) and free-living/planktonic community controls. Culture and molecular methods (quantitative polymerase chain reaction (qPCR)) were used to ascertain phenotypic and genotypic AMR prevalence, respectively, and multiplex colony PCR was used to identify extra-intestinal pathogenic Escherichia coli (ExPECs). From this, polystyrene and wood particles were found to significantly enrich AMR bacteria, whereas sewage-sourced bio-beads significantly enriched ExPECs. Polystyrene and wood were the least smooth particles, and so the importance of particle roughness on AMR prevalence was then directly investigated by comparing the colonization of virgin vs artificially weathered polyethylene particles. Surface weathering did not have a significant effect on the AMR prevalence of colonized particles. Our results suggest that the colonization of plastic and non-plastic particles by AMR and pathogenic bacteria may be enhanced by substrate-specific traits.

Antiviral Activity of Plant-Based Additives Against African Swine Fever Virus (ASFV) in Feed Ingredients.

African swine fever (ASF) is one of the deadliest swine diseases with haemorrhagic symptoms and a high mortality rate. Plant-derived additives are potential antiviral agents against viruses due to their environmental and user-friendly properties. This study aims to evaluate the efficacy of plant-based additives (Phyto.A04 and Phyto.B) compared to an organic acid blend (OAB) in inactivating ASF virus (ASFV) in cell culture and feed. ASFV-spiked feed was treated with individual or combined additives such as OAB, Phyto.A04 and Phyto.B. The viability of ASFV after treatment of ASFV-spiked feed with additives was then confirmed by both methods, real-time PCR and cell culture. The results of the in vitro test with cell cultures showed that all three additives (OAB, Phyto.A04 and Phyto.B) exerted a strong virucidal effect on ASFV in porcine alveolar macrophage cells. OAB at a concentration of 0.3% reduced the virus concentration from 4.48 log10 HAD50/mL after 1 day of treatment (day 1) to 3.29 log10 HAD50/mL after 3 days of treatment (day 3) and remained undetected after 7 days of treatment (day 7). In Phyto.A04 with 1%, the virus was only detectable on day 1 (3.53 log10 HAD50/mL). Phyto.B with 0.01% and 0.05% both showed good efficacy in completely inhibiting virus presence on days 3 and 7. All additives, OAB, Phyto.A04 and Phyto.B, were able to inactivate ASFV in a dose-dependent manner, as confirmed by cell culture and PCR methods. The combination of additives at different concentrations consistently improved the virucidal results.

Decomposition of Polythene by Bacteria Isolated from Waste Disposal Sites in Gusau City

The accumulation of polythene can be hazardous and can cause various environmental problems. This work was aimed at studying the biodegradation of polythene by bacteria isolated from waste disposal areas in North West, (Zamfara state) Nigeria. A total of twenty soil samples were collected from four different locations, such as waste disposal areas, hospitals, roadsides, and petrol pumps station. Spread plate method was carried out on Nutrient agar for the growth of bacteria. A total of 13 bacteria were isolated from different areas. The total bacteria count in hospital ranged from 80 × 10-4 - 248 × 10-4 cfu/mL, roadsides ranged from 116 × 10-4 - 208 × 10-4 cfu/mL, petrol pump ranged from 88 × 10-4 - 184 × 10-4 cfu/mL and waste disposal areas ranged from 116 × 10-4 - 264 × 10-4 cfu/mL. The microbial species associated with the polythene materials were identified as Pseudomonas sp, Bacillus sp, Staphylococcus aureus, Providencia stuartii, Clostridium perfringes, Corynebacterium diphtheria, Listeria monocytogen. The bacteria isolates were screened for polyethylene degradation using mineral salt medium containing polyethylene as sole source of carbon and nitrogen for their ability to degrade polyethylene for the period of 2 months. The efficacy of the microbes in the degradation of polythene were analyzed using liquid culture method, among the bacteria Pseudomonas aeruginosa degraded 75% of polythene, Bacillus subtilis 71.4% in a period of 2 months.

A Retrospective Analysis of Broad-Range Microbial PCR-Sequencing Findings and Culture Results in 101 Bronchoalveolar Lavage Specimens

Abstract Introduction/Objective Pneumonia is a significant cause of illness and death, identifying its cause is challenging and critical for effective antibiotic treatment and for reducing the length of stay. Bronchoalveolar lavage is commonly performed to obtain high-quality lower respiratory tract samples. While traditional culture methods have been the standard for pathogen detection, microbial rRNA broad-range polymerase chain reaction (BR-PCR) has emerged as an alternative method. Methods/Case Report A retrospective analysis was conducted on 101 patients suspected of severe pulmonary infections at Oregon Health and Science University (OHSU). Results (if a Case Study enter NA) When important microbial etiology is considered, culture detected 26%, while BR- PCR detected 20%. Both methods proved effective in detecting common pathogens; however, culture demonstrated superiority in detecting fungal pathogens due to potential sampling biases associated with PCR. Furthermore, culture holds the advantages of being faster and more cost-effective, although it may fail to detect difficult to non-cultivable organisms, like Legionella spp. Culture is indispensable for susceptibility testing and strain typing purposes. More often than not, BR-PCR captures airway microbiota indiscriminately with minimal clinical significance, potentially confusing clinical interpretation. Conclusion Novel technologies like Biofire Pneumonia Panel (BPP), a multiplex PCR, have shown promise in the rapid detection of pneumonia pathogens including bacteria, viruses, and resistance genes (< 2 hours), complementing the traditional culture methods for microbiological diagnosis. Based on this analysis we recommend introducing BPP at OHSU.

Assessment of Trichoderma Species Isolated From Volcanic Soil of a Durian Field in Sisaket Province, Thailand for Plant Growth Promotion and Biocontrol Potential

Trichoderma species are ubiquitous saprophytic fungi commonly found in soil, with potential as fungal biocontrol agents for plant disease control and growth promotion. This research aimed to assess the attributes of Trichoderma sp. as plant growth promoters, focusing on nitrogen fixation, siderophore production, plant nutrient solubilization and indole-3-acetic acid production. Out of 19 isolates tested, 6 (T05, T19, T25, T27, T28 and T33) showed superior plant growth promotion abilities. These isolates were further evaluated for their capacity to enhance the germination of Khao Dawk Mali 105 rice seeds. Seeds were soaked in a suspension of conidia at a concentration of 1×1012 conidia/mL, and germination rates were measured. The germination rate ranged between 77.67 - 86.67 %, statistically significantly higher than the control rate of 33.33 %. Among the isolates, T05 exhibited the highest promotion of plant growth, evidenced by root length (7.27 cm), shoot length (8.50 cm), fresh weight (0.54 g) and dry weight (0.27 g), all significantly different from the control. Additionally, the potential of isolates to inhibit the fungal pathogen Colletotrichum sp. MN-3 was assessed using a dual culture method, with T35 showing the highest inhibition percentage (60.90 %). Morphological classification and nucleotide sequencing of the ITS1-5.8S-ITS2 region of rDNA gene identified the isolates as T. harzianum, T. reesei, T. asperellum and T. longibrachiatum. These findings underscore the effectiveness of Trichoderma sp. in promoting plant growth and suppressing plant pathogenic fungi. HIGHLIGHTSSix Trichoderma isolates (T05, T19, T25, T27, T28 and T33) exhibited superior plant growth promotion abilities. These isolates significantly enhanced germination rates of Khao Dawk Mali 105 rice seeds compared to the method. Among the isolates, T05 showed the greatest promotion of plant growth, as evidenced by root length, shoot length, fresh weight and dry weight. Trichoderma isolates demonstrated potential for inhibiting the fungal pathogen Colletotrichum sp. MN-3. Morphological classification divided the Trichoderma sp. into 4 groups, and species identification through nucleotide sequencing revealed T. harzianum, T. reesei, T. asperellum and T. longibrachiatum. GRAPHICAL ABSTRACT

Influence of European Genre Model on Chinese Chamber Instrumental Ensemble with Piano

In today's world, the rapid development of Chinese culture and its influence on the music scene underscore the necessity for systematic research into the Chinese piano chamber ensemble. The growing interest in this musical genre challenges many widely accepted notions about musical tradition and prompts a deeper understanding of its characteristics and impact on contemporary music art. The purpose of the research is to study and analyse the musical and cultural interrelationships within the Chinese piano chamber ensemble to better understand its essence and role in the modern music world. The research methodology is based on a comprehensive approach, incorporating analysis of musical material, historical and cultural methods, as well as comparative analysis. Modern theoretical concepts are applied to reveal the significance of the piano chamber ensemble in the context of cultural transformation and to examine the interaction and influence of national and non-national musical traditions on the formation of the musical genre, as well as its role in cultural exchange and interaction between different cultural traditions. The scientific novelty of the research lies in the first attempt to systematise and analyse the musical-cultural interrelationships within the Chinese piano chamber ensemble, which opens up new horizons for further understanding of musical culture and processes of cultural exchange in the contemporary world. The conclusions of the research testify to the importance of the Chinese piano chamber ensemble in contemporary music culture and its potential as a phenomenon of musical art. It not only reflects the interrelationship between national and non-national musical traditions but also serves as a significant factor in cultural exchange and interaction between different cultural traditions.

Efficient Treatment of Psoriasis Using Conditioned Media from Mesenchymal Stem Cell Spheroids Cultured to Produce Transforming Growth Factor-β1-Enriched Small-Sized Extracellular Vesicles.

Psoriasis is a common chronic inflammatory disease in which keratinocytes proliferate abnormally due to excessive immune action. Psoriasis can be associated with various comorbidities and has a significant impact on health-related quality of life. Although many systemic treatments, including biologic agents, have been developed, topical treatment remains the main option for psoriasis management. Consequently, there is an urgent need to develop topical treatments with minimal side effects and high efficacy. Mesenchymal stem cells (MSCs) exhibit excellent immune regulation, anti-inflammatory activities, and therapeutic effects, and MSC-derived extracellular vesicles (EVs) can serve as crucial mediators of functional transfer from MSCs. Therefore, this study aimed to develop a safe and easy-to-use emulsion cream for treating psoriasis using MSC conditioned media (CM) containing EVs. We developed an enhanced Wharton's jelly MSC (WJ-MSC) culture method through a three-dimensional (3D) culture containing exogenous transforming growth factor-β3. Using the 3D culture system, we obtained CM from WJ-MSCs, which yielded a higher EV production compared to that of conventional WJ-MSC culture methods, and investigated the effect of EV-enriched 3D-WJ-MSC-CM cream on psoriasis-related inflammation. Administration of the EV-enriched 3D-WJ-MSC-CM cream significantly reduced erythema, thickness, and scaling of skin lesions, alleviated imiquimod-induced psoriasiform lesions in mice, and ameliorated histopathological changes in mouse skin. The upregulated mRNA expression of inflammatory cytokines, including IL-17a, IL-22, IL-23, and IL-36, decreased in the lesions. In conclusion, we present here a new topical treatment for psoriasis using an MSC EV-enriched cream.

Evolution of Flute Art: From Archaic Musical Traditions to the Present

The purpose of the work is to substantiate the evolution of pipe art as a special phenomenon from archaic musical traditions to the present. The methodology. Significant for this work were historical, analytical, musicological, and cultural methods, which were used to reveal various aspects of flute art, including its evolution, use in different cultural and social contexts, and its impact on the music culture of Ukraine. The scientific novelty of the work is determined by its analytical perspective, which represents a comprehensive view of the history and evolutionary development of pipe art from archaic musical traditions to the present. Conclusions. The presented materials reveal the evolution of pipe art and its role in the modern world of musical performance and musical genres. Attention is focused on changes in materials and construction when making a musical instrument. The advantages of the chromatic flute as a unique instrument with a wide range of performance and musical-tonal possibilities are argued. The study demonstrates the significance and relevance of flute art for contemporary cultural and musical environments, opening up new prospects for future research in musicology.

Effects of BYL-719 (alpelisib) on human breast cancer stem cells to overcome drug resistance in human breast cancer.

Breast cancer continues to be a major health concern and is currently the most commonly diagnosed cancer worldwide. Relapse, metastasis, and therapy resistance are major clinical issues that doctors need to address. We believe BYL-719, which is PI3 kinase p110а inhibitor, could also inhibit the breast cancer stem cell phenotype and epithelial-to-mesenchymal transition (EMT). In addition to the PI3K/AKT signaling pathway, BYL-719 can also inhibit essential cancer-related signaling pathways, all of which would ultimately act on the microenvironment of cancer stem cells, which is quite complicated and regulates the characteristics of tumors. These include the stemness and resistance of malignant tumors, plasticity of cancer stem cells, and anti-apoptotic features. A three-dimensional (3D) mammosphere culture method was used in vitro to culture and collect breast cancer stem cells (BCSCs). MTT, clonogenic, and cell apoptosis assays were used to detect cell viability, self-renewal, and differentiation abilities. A sphere formation assay under 3D conditions was used to detect the mammophore inhibition rate of BYL-719. The subpopulation of CD44+CD24- was detected using flow cytometry analysis while EMT biomarkers and essential signaling pathways were detected using western blotting. All the data were analyzed using GraphPad Prism 9 software. BCSC-like cells were obtained by using the 3D cell culture method in vitro. We confirmed that BYL-719 could inhibit BCSC-like cell proliferation in 3D cultures and that the stemness characteristics of BCSC-like cells were inhibited. The PI3K/AKT/mTOR signaling pathway could be inhibited by BYL-719, and the Notch, JAK-STAT and MAPK/ERK signaling pathways which have crosstalk in the tumor microenvironment (TME) are also inhibited. By comparing eribulin-resistant breast cancer cell lines, we confirmed that BYL-719 could effectively overcome drug resistance. The 3D cell culture is a novel and highly effective method for enriching BCSCs in vitro. Furthermore, the stemness and EMT of BCSCs were inhibited by BYL-719 by acting on various signaling pathways. Finally, we believe that drug resistance can be overcome by targeting the BCSCs. Conjugation of BYL-719 with other anti-neoplastic agents may be a promising treatment for this in clinic.

Clinical Utility of a Multiplex PCR Panel (BioFire Joint Infection®) in the Adjustment of Empiric Antimicrobial Therapy: Experience in Pediatric Osteoarticular Infections.

This study aims to evaluate the impact of the PCR multiplex panel (BioFire JI®) on the diagnosis and management of pediatric osteoarticular infections. This retrospective study analyzed data from pediatric patients diagnosed with osteoarticular infections between January 2023 and April 2024. The effectiveness of the PCR multiplex panel in identifying pathogens was compared with traditional culture methods. In total, 50 patients were identified (66.6% male, 74% under 3 years of age). They were diagnosed as follows: septic arthritis in 46%, osteomyelitis in 26%, and septic osteoarthritis in 22%. An identifiable agent was isolated by conventional culture in 22 cases (44%). Kingella kingae was the predominant pathogen identified, accounting for 50% of cases (11/22), followed by Staphylococcus aureus (9/22). The BioFire JI® Panel PCR demonstrated a sensitivity of 93%, with a specificity of 63% when evaluated against synovial fluid culture as the reference standard. The panel identified seven additional pathogens not detected by conventional culture methods: 2/9 MSSA (22%), 1/1 S. pyogenes (100%), and 4/11 K. kingae (37%), increasing the yield by 14%. The rapid identification of pathogens facilitated timely and targeted therapeutic interventions. The PCR multiplex panel (BioFire JI®) improved the diagnosis of pediatric osteoarticular infections.

Song Heritage of “The Beatles” in Academic Musical Interpretations

The purpose of the research. The paper reveals the features of the approaches of academic musicians to the creativity of “The Beatles” and characterises the musical projects created on the basis of the song heritage of the legendary British group. The research methodology consists in the application of historical cultural, comparative and musical analytical methods, which allowed to reveal the academic potential of “The Beatles” music and outline the ways and directions of academisation of their song creativity. The scientific novelty consists in the fact that for the first time academic interpretations of the songs of “The Beatles” group, carried out by L. Berio, J. Bayless and P. Breiner, were characterised, and two approaches to musical transcriptions of the song heritage of the group were outlined – academic and academicized, the type corresponds to all the parameters of academic music, the second gravitates towards classical crossover as a direction of pop music. Conclusions. Among the numerous cover versions, transcriptions and paraphrases of “The Beatles” music, a number of original musical interpretations are attractive, the authors of which turn to the Baroque style. The vocal cycle “Beatles Songs” by L. Berio (1967), the instrumental albums “Bach Meets the Beatles” (1984), “Bach Meets the Beatles. Revisited” (2004) by J. Bayless and “Beatles Go Baroque” (1993) by P. Breiner have already acquired the status of iconic. Their authors, who received a classical musical education, give priority to academic interpretive versions, where the songs of “The Beatles” group are considered as thematic material, which becomes the basis for academic type compositions with classical musical dramaturgy formed in the instrumental music of the Modern period. A number of compositions, in which the basic features of a song are preserved, gravitate towards academicized type interpretations, approaching a classical crossover, and can represent both academic and pop musical creativity.

Neonatal exposure to high d-galactose affects germ cell development in neonatal testes organ culture

Excess exogenous supplementation of d-galactose (d-gal), a monosaccharide and reducing sugar, generates reactive oxygen species (ROS), leading to cell damage and death. ROS accumulation is critical in aging. Therefore, d-gal-induced aging mouse models are used in aging studies. Herein, we evaluated d-gal’s effect on neonatal testis development using an in vitro organ culture method. Mouse testicular fragments (MTFs) derived from neonatal testes (postnatal day 5) were cultured with 500 mM d-gal for 5 days. d-gal-treated MTFs showed a significantly increased and decreased expression of undifferentiated and differentiated germ cell markers, respectively, with a substantial reduction in meiotic cells. In d-gal-exposed MTFs, expression levels of Sertoli cell markers (Sox9 and Wt1) increased, while those of StAR and 17β-HSD3, whose expressions are abundant in d-Gal treated adult Leydig cells, decreased. Additionally, the enzyme 3 β-HSD1, essential for steroidogenesis in Leydig cells, was significantly reduced in d-gal-exposed MTFs compared to that in controls.d-gal significantly increased the expression of Bad, Bax, and cleaved caspase-3 and -8. Via oxidative stress in MTF. Overall, d-gal negatively regulates germ cell and Leydig cell development in neonatal testes through pro-apoptotic mechanisms and ROS.

Identification Of Pathogen Causing Wilt Disease Of Chinese Yam (Dioscorea Polystachya Turczaninow) In Hunan

This study aims to identify the pathogen causing wilt disease of Chinese yam (Dioscorea polystachya Turczaninow.) in Hunan Province. Diseased stems were collected from the fields in Loudi City in the Hunan Province. Tissue culture method was used for the isolation of the pathogen causing wilt disease of Chinese yam, and was identified as Fusarium asiaticum based on the morphological characteristics and molecular technology. Out of five fungicides, Difenoconazole· Azoxystrobin 40% SC, Hexaconazole 5% SC, and Difenoconazole 10% WDG, demonstrated inhibitory effects on the pathogen. Among them, hexonazole 5% SC had the lowest EC50 value and was the most sensitive to the pathogen of Chinese yam wilt, so it should be selected first for the control of Chinese yam wilt. Bangladesh J. Bot. 53(3): 857-863, 2024 (September) Special

Genome sequencing analysis of the pncA, rpsA and panD genes responsible for pyrazinamide resistance of Mycobacterium tuberculosis from Indonesian isolates.

Developing the most suitable treatment against tuberculosis based on resistance profiles is imperative to effectively cure tuberculosis patients. Whole-genome sequencing is a molecular method that allows for the rapid and cost-effective detection of mutations in multiple genes associated with anti-tuberculosis drug resistance. This sequencing approach addresses the limitations of culture-based methods, which may not apply to certain anti-TB drugs, such as pyrazinamide, because of their specific culture medium requirements, potentially leading to biased resistance culture results. Thirty-four M. tuberculosis isolates were subcultured on a Lowenstein-Jensen medium. The genome of these bacteria was subsequently isolated using cetyltrimethylammonium bromide. Genome sequencing was performed with Novaseq Illumina 6000 (Illumina), and the data were analysed using the GenTB and Mykrobe applications. We also conducted a de novo analysis to compare the two methods and performed mutation analysis of other genes encoding pyrazinamide resistance, namely rpsA and panD. The results revealed mutations in the pncA gene, which were identified based on the databases accessed through GenTB and Mykrobe. Two discrepancies between the drug susceptibility testing and sequencing results may suggest potential instability in the drug susceptibility testing culture, specifically concerning PZA. Meanwhile, the results of the de novo analysis showed the same result of pncA mutation to the GenTB or Mykrobe; meanwhile, there were silent mutations in rpsA in several isolates and a point mutation; no mutations were found in the panD gene. However, the mutations in the genes encoding pyrazinamide require further and in-depth study to understand their relationship to the phenotypic profile. Compared to the conventional culture method, the whole-genome sequencing method has advantages in determining anti-tuberculosis resistance profiles, especially in reduced time and bias.

Zum Stand des Fachs Cultură și civilizație an rumänischen Universitäten

Abstract Culture and civilization is probably one of the most discussed areas in foreign language didactics. It is taught both implicitly in the teaching of German as a foreign language and as an independent subject in university teaching. It is also offered at many Romanian universities as part of German-language degree courses. Especially in subjects in which prospective teachers and translators are trained, culture and civilization occupies a special position because it plays the undisputedly important role of imparting cultural knowledge about the target language country, which is seen as the second component in foreign language acquisition alongside linguistic knowledge. While foreign language lessons usually deal with everyday cultural topics, university courses on culture and civilization tend to focus on geography, history, politics, media, art and/or economics, usually according to the respective focus of the department where the culture and civilization course is offered. This article deals with academic research on the subject of culture and civilization at Romanian universities after 1995, specifically with the objectives and methods for teaching culture and civilization, the organization of encounters in culture and civilization lessons and possible subject catalogs.

Development of Haploid Plants by Shed-Microspore Culture in Platycodon grandiflorum (Jacq.) A. DC.

Anther and microspore cultures are efficient methods for inducing haploids in plants. The microspore culture by chromosome-doubling method can produce double haploid lines, developing pure lines within the first or second generations. This study aimed to induce haploid plants in Platycodon grandiflorum using the shed-microspore culture method. P. grandiflorum floral buds (n = 1503) were cultured in six types of medium to induce haploids. Anthers were placed on a solid-liquid double-layer medium and cold pre-treated at 9 °C for one week, followed by incubation in the dark at 25 °C. Embryogenesis was observed after approximately 70 days of culture, producing haploid plants through regeneration. Of the 1503 floral buds, embryos developed in 120 buds, resulting in the induction of 402 individuals. Among the media used, Schenk and Hildebrandt (SH) and 1/2SH exhibited high efficiency, with embryogenesis ratios of 12% and 13.4%, respectively. Additionally, the highest embryogenesis ratio (15.3%) was observed in flower buds sized 10 mm or less. Therefore, we established shed-microspore culture conditions to induce haploids in P. grandiflorum. Using this method, haploids can be efficiently induced in P. grandiflorum, shortening the breeding period by enabling the rapid development of inbred lines.

Characterizing the urobiome in geriatric males with chronic indwelling urinary catheters: an exploratory longitudinal study.

The impact of chronic indwelling urinary catheters (IUCs) on the composition and stability of the urinary microbiota remains unknown. The primary aim of this study was to describe the urinary microbiomes of geriatric males with chronic IUCs. A secondary aim was to explore clinical catheter-associated urinary tract infection (CAUTI) courses of the participants. Geriatric male patients with chronic IUCs were followed longitudinally. Catheterized urine, catheter tips, and both urethral and periurethral swabs were collected from participants at monthly intervals. Microbes were isolated and identified from each specimen using an enhanced culture method called expanded quantitative urine culture (EQUC) and targeted 16S rRNA gene DNA sequencing. Microbial outcomes were examined both in the absence of urinary symptoms and in the context of clinical diagnosis of CAUTI. Ten male participants (mean age 86 years) were enrolled. Urinary microbiomes differed for each participant. However, within each individual, microbiomes were similar over time and across niches (bladder, catheter, urethra, and periurethra). Within-niche microbiomes differed across individuals, and this was observed over time. The most abundant bacteria isolated from all niches were known uropathogens. Six of 10 individuals met diagnostic criteria for CAUTI at least once during the 12-month observation period, but no evidence of this or antibiotic treatment/response was discernable in our monthly samples. The microbiomes of each participant were unique and remained similar over time and across niches. Longitudinal EQUC or 16S rRNA gene sequencing data could be useful to clinicians when diagnosing or treating possible CAUTI.IMPORTANCECatheter-associated urinary tract infections (CAUTIs) are serious but preventable nosocomial infections. The most common risk factor for developing CAUTI is prolonged use of indwelling urinary catheters (IUCs). This study provides the first longitudinal description of the urinary microbiomes of geriatric males with chronic IUCs, in the absence of urinary signs and symptoms, as a first step toward enhancing our knowledge of the impact of chronic IUCs on the composition and stability of the urinary microbiota. This is an understudied area, particularly for males.