Culture Methods Research Articles

Phytochemical Riches, Bioactivities, and Clinical Potential of Prinsepia utilis Royle: A Comprehensive Review

Prinsepia utilis Royle, widely recognized in Chinese and Indian traditional medicine, has attracted significant interest in the realms of phytochemistry and bioactivity research. This deciduous shrub from the Rosaceae family thrives in the Himalayan region at altitudes between 1000 and 3000 meters. The current paper aims to consolidate existing knowledge on P. utilis, providing an updated database of its phytochemical constituents and related bioactivities. Investigations have revealed a wealth of secondary metabolites, including alkaloids, flavonoids, phenolics, and terpenoids. Research has shown that extracts, fractions, and isolated compounds of P. utilis exhibit notable antioxidant, anti‐inflammatory, antimicrobial, enzymatic inhibitory, immunosuppressive, and cytotoxic properties. In vivo studies confirm its traditional uses, demonstrating anti‐inflammatory, anti‐benign prostatic hyperplasia, osteoprotective, and hypoglycemic effects. Effective formulations using P. utilis extract have been developed for atrophic dermatitis and acne vulgaris. These findings underscore the plant's potential as a source of new therapeutic agents and functional additives. Nonetheless, further research is essential to advance pharmacological assessments, clinical trials, and sustainable cultivation methods to fully harness its pharmaceutical and industrial applications.

Effects of microgravity on human iPSC-derived neural organoids on the International Space Station.

Research conducted on the International Space Station (ISS) in low-Earth orbit (LEO) has shown the effects of microgravity on multiple organs. To investigate the effects of microgravity on the central nervous system, we developed a unique organoid strategy for modeling specific regions of the brain that are affected by neurodegenerative diseases. We generated 3-dimensional human neural organoids from induced pluripotent stem cells (iPSCs) derived from individuals affected by primary progressive multiple sclerosis (PPMS)or Parkinson's disease (PD) and non-symptomatic controls, by differentiating them toward cortical and dopaminergic fates, respectively, and combined them with isogenic microglia. The organoids were cultured for a month using a novel sealed cryovial culture method on the International Space Station (ISS) and a parallel set that remained on Earth. Live samples were returned to Earth for analysis by RNA expression and histology and were attached to culture dishes to enable neurite outgrowth. Our results show that both cortical and dopaminergic organoids cultured in LEO had lower levels of genes associated with cell proliferation and higher levels of maturation-associated genes, suggesting that the cells matured more quickly in LEO. This study is continuing with several more missions in order to understand the mechanisms underlying accelerated maturation and to investigate other neurological diseases. Our goal is to make use of the opportunity to study neural cells in LEO to better understand and treat neurodegenerative disease on Earth and to help ameliorate potentially adverse neurological effects of space travel.

Unveiling the Human Brain on a Chip: An Odyssey to Reconstitute Neuronal Ensembles and Explore Plausible Applications in Neuroscience.

The brain is an incredibly complex structure that consists of millions of neural networks. In developmental and cellular neuroscience, probing the highly complex dynamics of the brain remains a challenge. Furthermore, deciphering how several cues can influence neuronal growth and its interactions with different brain cell types (such as astrocytes and microglia) is also a formidable task. Traditional in vitro macroscopic cell culture techniques offer simple and straightforward methods. However, they often fall short of providing insights into the complex phenomena of neuronal network formation and the relevant microenvironments. To circumvent the drawbacks of conventional cell culture methods, recent advancements in the development of microfluidic device-based microplatforms have emerged as promising alternatives. Microfluidic devices enable precise spatiotemporal control over compartmentalized cell cultures. This feature facilitates researchers in reconstituting the intricacies of the neuronal cytoarchitecture within a regulated environment. Therefore, in this review, we focus primarily on modeling neuronal development in a microfluidic device and the various strategies that researchers have adopted to mimic neurogenesis on a chip. Additionally, we have presented an overview of the application of brain-on-chip models for the recapitulation of the blood-brain barrier and neurodegenerative diseases, followed by subsequent high-throughput drug screening. These lab-on-a-chip technologies have tremendous potential to mimic the brain on a chip, providing valuable insights into fundamental brain processes. The brain-on-chip models will also serve as innovative platforms for developing novel neurotherapeutics to address several neurological disorders.

Unravelling the complexity of bovine milk microbiome: insights into mastitis through enterotyping using full-length 16S-metabarcoding

BackgroundMastitis, inflammation of the mammary gland, is a major disease of dairy cattle and the main cause for antimicrobial use. Although mainly caused by bacterial infections, the aetiological agent often remains unidentified by conventional microbiological culture methods. The aim of this study was to test whether shifts in the bovine mammary gland microbiota can result in initiation or progression of mastitis.MethodsOxford-Nanopore long-read sequencing was used to generate full-length 16S rRNA gene reads (16S-metabarcoding) to characterise the microbial population of milk from healthy and diseased udder of cows classified into five groups based on their mastitis history and parity.ResultsSamples were classified into six enterotypes, each characterised by a marker genus and several differentially-abundant genera. Two enterotypes were exclusively composed of clinical mastitis samples and displayed a marked dysbiosis, with a single pathogenic genus predominating and displacing the endogenous bacterial population. Other mastitis samples (all subclinical and half of the clinical) clustered with those from healthy animals into three enterotypes, probably reflecting intermediate states between health and disease. After an episode of clinical mastitis, clinical recovery and microbiome reconstitution do not always occur in parallel, indicating that the clinical definition of the udder health status does not consistently reflect the microbial profile.ConclusionsThese results show that mastitis is a dynamic process in which the udder microbiota constantly changes, highlighting the complexity of defining a unique microbiota profile indicative of mastitis.

Formation of treatment regimens in newly diagnosed patients before the diagnosis of pulmonary tuberculosis verification

Abstract. One of the basic principles of treatment of tuberculosis patients is the appointment of treatment taking into account the drug resistance of Mycobacterium tuberculosis (MBT). However, during treatment, it may be necessary to change the intake regimen when new data on drug resistance become available. The search for ways to choose the optimal treatment method seems relevant.Aim: To analyze the prescribed regimens and the reason for their correction in newly identified patients before verifying the diagnosis of tuberculosis.Material and Methods. The following methods were used to detect MBT: luminescent microscopy, molecular genetic methods and seeding on liquid and solid media. Tests for drug sensitivity were conducted in parallel. The study group consisted of patients with established bacterial excretion (n = 79), whose diagnosis was confirmed by cultural methods. A retrospective method was used in the analysis.Results. At the beginning of treatment with bacterioscopy and polymerase chain reaction methods MBT was detected in 65/79 (82.2%) cases, the remaining 14 cases were detected only by culture methods. Forced change of the therapy regimen during treatment after receiving the results was in 25/79 (31.6%) cases, of which 1/25 (4%) with isoniazid resistance, 7/25 (28%) with multidrug-resistant tuberculosis, 17/25 (68%) with the pre-extensive drug resistance regimen. The latter was prescribed only after receiving results of tests for drug sensitivity on liquid and solid media after 1–3 months. The multidrug-resistant tuberculosis treatment regimen decreased after receiving data on resistance to fluoroquinolones in accordance with the seeding data, but the largest number of patients still remained on this regimen – 41/79 (51.9%). Among these patients a change in chemotherapy regimen from multidrug-resistant tuberculosis to pre-extensive drug resistance during the intensive phase of treatment occurred in 17/25 (68%).Conclusion. When prescribing a regimen, it is recommended to assess the risk of changing the regimen and, if there are predictors, apply a chemotherapy regimen before treatment.

Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels Under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions.

Mesenchymal stem cells (MSCs) hold immense therapeutic potential due to their regenerative and immunomodulatory properties. However, to utilize this potential, it is crucial to optimize their in vitro cultivation conditions. Three-dimensional (3D) culture methods using cell-laden hydrogels aim to mimic the physiological microenvironment in vitro, thus preserving MSC biological functionalities. Cellulosic hydrogels are particularly promising due to their biocompatibility, sustainability, and tunability in terms of chemical, morphological, and mechanical properties. This study investigated the impact of (1) two physical crosslinking scenarios for hydrogels derived from anionic cellulose nanofibers (to-CNF) used to encapsulate adipose-derived MSCs (adMSCs) and (2) physiological culture conditions on the in vitro proliferation, differentiation, and extracellular vesicle (EV) production of these adMSCs. The results revealed that additional Ca2+-mediated crosslinking, intended to complement the self-assembly and gelation of aqueous to-CNF in the adMSC cultivation medium, adversely affected both the mechanical properties of the hydrogel spheres and the growth of the encapsulated cells. However, cultivation under dynamic and hypoxic conditions significantly improved the proliferation and differentiation of the encapsulated adMSCs. Furthermore, it was demonstrated that the adMSCs in the CNF hydrogel spheres exhibited potential for scalable EV production with potent immunosuppressive capacities in a bioreactor system. These findings underscore the importance of physiological culture conditions and the suitability of cellulosic materials for enhancing the therapeutic potential of MSCs. Overall, this study provides valuable insights for optimizing the in vitro cultivation of MSCs for various applications, including tissue engineering, drug testing, and EV-based therapies.

Research on the Multilingual Talent Cultivation System Empowered by AI under the New Liberal Arts Context

This paper explores the AI (artificial intelligence) empowerment in foreign language education within the context of “New Liberal Arts” and its impact on a diversified foreign language talent cultivation system. Though challenges brought by AI, AI empowerment could facilitate the foreign language discipline development. Drawing upon this basis, the paper further explores the breakthroughs in foreign language talent cultivation. That suggests a shift from information tools to character education, from language proficiency to core competency strengthening, from single-discipline to interdisciplinary talent cultivation, and from imparting language knowledge to advancing general education understanding. This paper proposes paths for diversified foreign language talent cultivation in the AI era, including AI empowerment character education cultivation approaches, the construction of a diversified curriculum system, the innovation of interdisciplinary curriculum clusters, AI empowerment reshaping of digital teaching practice settings, and AI empowerment reconstruction of foreign language digital teaching resources. With those approaches, the adjusting focus and methods of foreign language talents cultivation, an effective Multilingual Talent Cultivation System will be established with international perspectives, cross-cultural communication skills, and innovation capabilities to meet the development needs of the AI era.

From Third Place: Illustrations and Observations of Connection in the Inner City

From Third Place is a practice-led project that explores the concept of third place in relation to society of the inner city, specifically Auckland CBD, using the traditional cultural anthropological methods of fieldwork and observational drawing to explore third places in Auckland. This project addresses the concept of third place and its role within society and the individual, reflecting on how we connect within our environment. This project is heavily influenced by my previous degree in cultural anthropology and art history. The inquiry is explored through the publication in three chapters of observational illustrations which focuses on the main third places used in everyday life by residents of Auckland CBD; cafes, parks, and pubs. The resulting publication is a series of snapshots of life in the city. The aims of this project is to explore connection of individuals in the city and celebrate the third places that facilitate them.

Research on the application of cultural communication teaching in college English class

This study explores the application of cultural communication method in English classroom teaching, aiming at improving students' intercultural communication ability and English language skills. The cultural communication approach emphasizes the key role of culture in language learning and helps students better understand and use English by teaching cultural background, social etiquette and rules of language use. Through practical application of CIT, this study examines its effectiveness in English teaching and explores students' feedback. The results show that CIT can help improve students' cultural sensitivity and communicative competence, and also promote the improvement of English language skills.

Optimization of Potato Cultivation Through the Use of Biostimulator Supporter

Seed potato treatment is vital for plant protection, yield enhancement, and product quality. In the conducted research, the plant biostimulator Supporter was applied to evaluate its impact on potato yields and its structure. Supporter contains both synthetic and SL amino acids, which promote plant growth by enhancing nutrient utilization and fostering the development of a more effective root system. Such a formulation allows to maintain better resistance to environmental stresses, which may include drought or nutrient deficiency, among others. The field study was conducted in 2015–2017 in four towns located in different regions of Poland (Barankowo, Głubczyce, Kędrzyno, and Ryn) using a randomized complete block design with a split-plot design. Varieties (‘Innovator’, ‘Lilly’, ‘Lady Claire’, and ‘Verdi’) were tested. The experiment compared the cultivation technology using Supporter biostimulator with which seed potatoes were treated compared to conventional cultivation (control object) by soaking the tubers in distilled water before planting. The total yield of potato tubers after Supporter application was higher by 13.3%, while the commercial yield increased by 21.1% compared to the traditional cultivation method. The most productive, regardless of cultivation technology and years of research, in terms of total tuber yield was the ‘Lilly’ variety with an average yield of 47.95 t∙ha−1, while the least productive variety was the ‘Innovator’ variety with an average yield of 29.93 t∙ha−1. The ‘Lady Claire’ variety had the highest commercial tuber yield, while the ‘Innovator’ variety had the lowest.

Spheroid culture to select theoretical therapeutic drugs in intravascular large B-cell lymphoma.

Intravascular large B-cell lymphoma (IVLBCL) is a rare type of extranodal large B-cell lymphoma that is characterized by the proliferation of lymphoma cells in the lumina of small vessels. Recent progress uncovering the genetic characteristics associated with MYD88/CD79B mutations has stimulated interest in the use of drugs targeting B-cell receptor signaling, including Bruton's tyrosine kinase. However, difficulties in culturing exvivo IVLBCL cells has hampered research on the development of novel therapies. In the present study, we demonstrated the establishment of an exvivo culture system of IVLBCL cells obtained from patient-derived xenograft (PDX) models. The spheroid culture enabled us to culture IVLBCL PDX cells for more than 10 days and to explore the efficacy of drug treatments acting on these cells. We found that carfilzomib and ibrutinib were effective for treating IVLBCL in exvivo experiments and conducted invivo analyses to assess the efficacy of these drugs. Although the efficacy of carfilzomib was difficult to confirm due to its toxicity in our models, ibrutinib showed comparable efficacy to a standard combination of chemotherapy drugs. Together, our data provide a new culture method for IVLBCL PDX cells and a rationale for translating ibrutinib to clinical use in IVLBCL patients.

Alternative Dispute Resolution: Historical, Customary Practices & Legal Aspects of Nepal

This paper focuses on the use of ADR in Nepal based on analyzing the historical background, cultural practices, and existing laws. The research's main objective was to identify how the ADR is practiced in the communities and in the judicial system of Nepal. The basic approach of research was doctrinal and in this approach primary sources regarded the constitution of Nepal and Acts likewise secondary source was taken as published articles and reports. It starts with background information on what peaceful conflict resolution is and how disputes are settled by mediators such as the elders and village assemblies conducted in communities. These are cultural methods of solving conflicts and they have been in use before the legal structures were formulated in Nepal. The paper also discusses the different forms of traditional ADR that have been incorporated into the contemporary Legal System of Nepal. It also surveys prominent statutes such as the Arbitration Act and the Mediation Act and how these laws assimilate and police traditional practices into and within the legal process. More specifically, the study compares the various conventional techniques, viewpoints, and designs used to address the issues of compliance with customary practices and measures the extent of the effectiveness. The paper nonetheless seeks to contrast between the traditional approaches and the legal frameworks to illustrate the need to embrace cultural practices alongside access to justice. The research found that the use of ADR is in different sectors of the communities and organizations, including international trade disputes and the law of Nepal has opened to settle disputes outside of the court system

Prevalence of Campylobacter and antibiotic susceptibility in chickens at slaughterhouses and retail markets in the Mekong Delta

This study aims to determine the prevalence of Campylobacter and its antibiotic susceptibility at slaughterhouses and live poultry markets in the Mekong Delta, Vietnam. A total of 177 samples, including 112 fecal samples, 28 cecal samples, and 37 rectal swab samples were collected. Thirty-nine samples tested positive for Campylobacter spp. by culture method, accounting for 22%. Isolation rates of Campylobacter in rectal swab samples were highest at 29.7% (11/37), followed by cecal samples at 25% (7/28) and fecal dropping samples at 18.8% (21/112). Species identification was performed by multiplex PCR and the result showed that the prevalence of Campylobacter coli (12.4%) was higher than Campylobacter jejuni (3.4%). Antibiotic susceptibility test showed that all isolated Campylobacter were susceptible to imipenem and meropenem (100%). Most Campylobacter were resistant to tetracycline (96.4%), ciprofloxacin (92.9%), nalidixic acid (89.3%), and ampicillin (67.9%). This study highlights the high contamination of Campylobacter and its antibiotic resistance in poultry in the Mekong Delta, Vietnam.

Insect Production: A Circular Economy Strategy in Iceland

In this review, the multifaceted issue of food security is addressed, emphasizing the need for innovative and culturally appropriate solutions. Exploring insect livestock farming emerges as a potential remedy, offering a pathway to alleviate food insecurity and promote food sovereignty, particularly when integrated with social acceptability. Stakeholder engagement on both production and consumption fronts, coupled with sustained support, is vital for successful implementation. The expanding landscape of commercial insect farming in the West prompts questions about its broader scalability and equitable deployment, especially for vulnerable populations. Existing research gaps underscore the need for a coordinated effort across international, national, and legal frameworks to effectively integrate insect farming into existing agricultural systems. In this review, we have delved into the industrial-scale production processes of mealworms and black soldier flies (BSFs), known for their high protein content and organic waste conversion, covering small and industrial cultivation methods, offering insights into mealworm production life cycles, innovative rearing systems, and harvesting techniques. This review concludes with climate-specific recommendations for insect facilities, stressing the importance of sustainable practices, continuous research and development, effective market strategies and economic feasibilities in Iceland. In the context of escalating demand for sustainable protein sources, industrial-scale insect production emerges as a pivotal player in addressing global food security challenges.

The productivity of selected soybean cultivars grown using various cultivation methods

Soybean is an important legume crop globally due to its rich protein, oil content, and functional components. The purpose of this study was to evaluate the yield of selected soybean cultivars depending on cultivation methods. The three-year field experiment, conducted from 2018 to 2020 at the Agricultural Experimental Station in Kępa-Puławy, Osiny farm (Institute of Soil Science and Plant Cultivation – State Research Institute in Puławy), investigated these variations. The first experimental factor was the soil cultivation method: A – conventional tillage, B – reduced tillage, and C – strip tillage. The second variable was soybean cultivar: ‘Aldana’ and ‘Merlin’. The soybean cultivars were selected for their differing maturity rates: ‘Aldana’ (000) is an early cultivar, while ‘Merlin’ (000++) semi-late cultivar. The field experiment utilised a split-plot design on Luvisol soil with sandy loam texture, belonging to a good rye complex, class IIIb–IVa, and was replicated four times. The study showed that the productivity (seed and protein yield) of the ‘Merlin’ cultivar grown in the central-eastern part of Poland was approximately 8% higher than that of the ‘Aldana’ cultivar. The cultivation method had a relatively minor influence on soybean yield, the content of selected nutrients, morphological features, and elements of the yield structure. The soil in strip-tillage method was more compact than the soil cultivated with a plough. After harvesting soybeans at a depth of 30, and 40 cm, the compactness of soil in strip-tillage or with reduced tillage was much lower than in spring, highlighting a positive effect of soybean cultivation on loosening the arable layer.

Water Use Efficiency in Rice Under Alternative Wetting and Drying Technique Using Energy Balance Model with UAV Information and AquaCrop in Lambayeque, Peru

In the context of global warming, rising air temperatures are increasing evapotranspiration (ETc) in all agricultural crops, including rice, a staple food worldwide. Simultaneously, the occurrence of droughts is reducing water availability, affecting traditional irrigation methods for rice cultivation (flood irrigation). The objective of this study was to determine ETc (water use) and yield performance in rice crop under different irrigation regimes: treatments with continuous flood irrigation (CF) and irrigations with alternating wetting and drying (AWD5, AWD10, and AWD20) in an experimental area in INIA–Vista Florida. Water balance, rice physiological data, and yield were measured in the field, and local weather data and thermal and multispectral images were collected with a meteorological station and a UAV (a total of 13 flights). ETc values obtained by applying the METRICTM (Mapping Evapotranspiration at High Resolution using Internalized Calibration) energy balance model ranged from 2.4 to 8.9 mm d−1 for the AWD and CF irrigation regimes. In addition, ETc was estimated by a water balance using the AquaCrop model, previously parameterized with RGB image data and field weather data, soil, irrigation water, and crops, obtaining values between 4.3 and 7.1 mm d−1 for the AWD and CF irrigation regimes. The results indicated that AWD irrigation allows for water savings of 27 to 28%, although it entails a yield reduction of from 2 to 15%, which translates into an increase in water use efficiency (WUE) of from 18 to 36%, allowing for optimizing water use and improving irrigation management.

Isolation, identification, and technological characteristics analysis of yeast strains from Pyracantha fortuneana fruits fermentation liquid.

Pyracantha fortuneana (P. fortuneana), as a medicinal and edible plant resource, is rich in nutrients. In order to screen the high quality yeast used in Firebone fruit wine, 12 strains of yeast were isolated and purified from P. fortuneana fermentation broth by traditional pure culture method. They were identified by molecular biology as Pichia kudriavzevii (P. kudriavzevii) and Saccharomyces cerevisiae (S. cerevisiae), respectively. Strain HJ-2 could grow normally at 30℃, alcohol content 15%, SO2 mass concentration 360mg/L, pH 3.2, sucrose mass concentration 400g/L and glucose mass concentration 250g/L. Strain HJ-6 could grow normally at 30℃, alcohol content 3%, SO2 concentration 360mg/L, pH 3.2, sucrose concentration 250g/L, glucose concentration 300g/L. Based on the technological characteristics of fruit wine, S. cerevisiae HJ-2 has the potential of brewing P. fortuneana fruit wine. P. kudriavzevii HJ-6 has a low tolerance to ethanol and is suitable for the production of fermented beverages such as low-alcohol wine or beer.

Relationships between fecal indicator abundance in water and sand and the presence of pathogenic genes in sand of recreational beaches.

For decades, the risk of exposure to infectious diseases in recreational beaches has been evaluated through the quantification of fecal indicator bacteria in water samples using culture methods. The analyses of sand samples have recently been developed as a complement to the monitoring of recreational waters in beach quality assessments. The growing use of molecular techniques for environmental monitoring allows for the rapid detection of pathogenic genes, thus providing more accurate information regarding the health risk of exposure to contaminated sand. The aim of this work was to determine the relationship between the fecal indicators abundance in water and sand and the presence of Shiga toxin-producer Escherichia coli (STEC) in sand by analyzing samples from touristic beaches using culture-dependent (fecal coliforms assay) and culture-independent (real-time PCR of stx1, stx2, and eae genes) techniques. We found a high concentration of coliform bacteria in water and sand in several beaches in eastern Uruguay, with different levels of sanitation networks and levels of urbanization. The presence of STEC virulence genes (mainly stx1) was confirmed in 8 out of 20 sand samples. The recreational use of sandy beaches may imply a risk to the health of its users, especially near streams and creek outflows, thus highlighting the need of monitoring sand bacteriological quality and pathogens using molecular tools.

The chronic wound characterisation study and biobank: a study protocol for a prospective observational cohort investigation of bacterial community composition, inflammatory responses and wound-healing trajectories in non-healing wounds

IntroductionChronic wounds affect 1%–2% of the global population, with rising incidence due to ageing and lifestyle-related diseases. Bacterial biofilms, found in 80% of chronic wounds, and scattered single-cell bacteria may...

Evaluation of the Fertilizer Potential of Black Soldier Fly Larval Frass Obtained by the Processing of Multiple Organic Wastes in Agriculture in Cameroon: Effects on Plant Growth Parameters and Maize Kernel Nutrient Composition

The growing utilization of inorganic fertilizers results in the depletion of a minimal quantity of essential elements, including phosphorus, nitrogen, and potassium. The residual fraction is discharged into the environment as organic waste, posing a threat to the ecosystem.The black soldier fly larvae (BSFL) have been acknowledged as a solution for the increasing problem of organic waste. BSFL provides numerous benefits, including their current use of waste material, known as frass, as an organic fertilizer. The aim of this study is to examine the fertilizing properties of frass and its effects on plant growth parameters and nutrient concentration in maize kernels when consumed. This will be compared to other cultivation methods including synthetic fertilizers, conventional compost, and no compost. Following a 90-day timeframe, a comprehensive analysis was performed to assess the growth characteristics and nutrient composition within the kennels. The results of this study indicate that frass positively affects plant growth and has a favorable blend of nutrients, as previously demonstrated by studies investigating the nutrient levels in maize. This study showcased the capacity of utilizing frass as a carrier to boost agricultural output and enhance the accessibility of particular nutrients in plants. Further investigation is required to substantiate these conclusions. The widespread use of black soldier fly larvae in the agricultural sector in poor countries, particularly in Sub-Saharan Africa, has the capacity to have a beneficial effect on integrated sustainable agriculture and the achievement of Sustainable Development Goals 2, 11, and 12.