Culture Period Research Articles

Effects of Cleaning Interval, Hapa Layers and Stocking Density on Growth Performance of Sandfish Holothuria scabra Juveniles in Ocean‐Based Nursery Production Systems

ABSTRACTThe mass production of release‐size H. scabra juveniles in the Philippines is primarily done in a floating hapa ocean‐based nursery systems. To optimize its production, understanding the influence of different culture conditions and their management is necessary. In this study, the effects of hapa net cleaning interval, hapa layers, and stocking density on growth and survival of sandfish juveniles reared in floating hapa (L × W × H: 1.7 m × 0.9 m × 1.2 m) nursery system were assessed in two 60‐day field studies. Results showed that the number of hapa net layers and cleaning interval had negligible effects on the growth performance of sandfish juveniles by day 30. However, sandfish juvenile survival was affected by hapa net layers with higher survival in triple‐layer mosquito net hapas (40.5 ± 0.97%) compared to double‐layer mosquito net hapas (17.8 ± 0.62%). By day 60, the growth performance of sandfish juveniles in double‐layer hapas was higher (average weight: 5.0 ± 0.97 g; AGR: 0.13 ± 0.03 g day−1) compared to that in triple‐layer mosquito hapa nets (average weight: 3.1 ± 0.23 g; AGR: 0.07 ± 0.01 g day−1). Survival of sandfish juveniles reared in double‐layer mosquito net hapas, cleaned at 5‐day intervals (85.9 ± 4.58%) was significantly lower compared to all other treatments. Conversely, initial juvenile stocking density did not significantly affect the growth performance and survival of sandfish juveniles in both rearing cycles. We therefore recommend the use of triple‐layer mosquito net hapas during the first 30‐day culture period and an initial stocking density of 1000 post‐settled sandfish juveniles to increase production output. After 30 days, when the juveniles are larger, the use of double‐layer mosquito net hapas and lower stocking density are recommended to reduce cost and enhance growth performance. Where cleaning is necessary, hapa nets may be cleaned at 10‐day intervals to improve survival. The results of this study have important implications for optimizing the production of release‐size sandfish in ocean‐based nursery systems in the Philippines.

Accurately Segmenting/Mapping Tobacco Seedlings Using UAV RGB Images Collected from Different Geomorphic Zones and Different Semantic Segmentation Models

The tobacco seedling stage is a crucial period for tobacco cultivation. Accurately extracting tobacco seedlings from satellite images can effectively assist farmers in replanting, precise fertilization, and subsequent yield estimation. However, in complex Karst mountainous areas, it is extremely challenging to accurately segment tobacco plants due to a variety of factors, such as the topography, the planting environment, and difficulties in obtaining high-resolution image data. Therefore, this study explores an accurate segmentation model for detecting tobacco seedlings from UAV RGB images across various geomorphic partitions, including dam and hilly areas. It explores a family of tobacco plant seedling segmentation networks, namely, U-Net, U-Net++, Linknet, PSPNet, MAnet, FPN, PAN, and DeepLabV3+, using the Hill Seedling Tobacco Dataset (HSTD), the Dam Area Seedling Tobacco Dataset (DASTD), and the Hilly Dam Area Seedling Tobacco Dataset (H-DASTD) for model training. To validate the performance of the semantic segmentation models for crop segmentation in the complex cropping environments of Karst mountainous areas, this study compares and analyzes the predicted results with the manually labeled true values. The results show that: (1) the accuracy of the models in segmenting tobacco seedling plants in the dam area is much higher than that in the hilly area, with the mean values of mIoU, PA, Precision, Recall, and the Kappa Coefficient reaching 87%, 97%, 91%, 85%, and 0.81 in the dam area and 81%, 97%, 72%, 73%, and 0.73 in the hilly area, respectively; (2) The segmentation accuracies of the models differ significantly across different geomorphological zones; the U-Net segmentation results are optimal for the dam area, with higher values of mIoU (93.83%), PA (98.83%), Precision (93.27%), Recall (96.24%), and the Kappa Coefficient (0.9440) than those of the other models; in the hilly area, the U-Net++ segmentation performance is better than that of the other models, with mIoU and PA of 84.17% and 98.56%, respectively; (3) The diversity of tobacco seedling samples affects the model segmentation accuracy, as shown by the Kappa Coefficient, with H-DASTD (0.901) > DASTD (0.885) > HSTD (0.726); (4) With regard to the factors affecting missed segregation, although the factors affecting the dam area and the hilly area are different, the main factors are small tobacco plants (STPs) and weeds for both areas. This study shows that the accurate segmentation of tobacco plant seedlings in dam and hilly areas based on UAV RGB images and semantic segmentation models can be achieved, thereby providing new ideas and technical support for accurate crop segmentation in Karst mountainous areas.

Economic Impact of Periphyton-Based Stunted Fingerling Production in Arunachal Pradesh, India

Non availability of right type quality seed at right time in required quantity is major bottleneck for the growth of aquaculture in Arunachal and major portion of seed are being import from other nearby state especially Assam in particular. To make availability of good quality seed at right time having a regional acclimation for a longer duration in far flung areas of the state, the present investigation was conducted on low cost Periphyton (grown on paddy straw) based stunted yearling production technology to improve the survival and health of stocked material and their economic impact on the fish farmers of district West Kameng during 2018-2021. The average values of pond water quality parameters were observed to vary between both the culture ponds with a significant seasonal differences at all the sites and there were found a greater difference in average survival rate of both the commodities i.e. table size fish as well as stunted yearling during whole culture period. The involved cost of cultivation was recorded higher Rs. 4,70,000 / ha in stunted yearling culture in comparison to Rs.424,940/ha in thetable size carp culture while average gross benefit was just opposite, recorded higher Rs. 6,90,000/ ha in stunted yearling culture in comparison to Rs.199,960/ ha in table size carp culture with the same trend in BCR i.e. 2.46 for stunted fingerling production and 1.47 in the production of table size fishes respectively.

Ethnic fermented foods of the world: an overview

AbstractThe origins of traditional fermented foods and the types and characteristics of fermented foods by region are reviewed. Fermented foods are classified into alcohol fermentation, acid fermentation, carbon dioxide (bread) fermentation, and amino acid/peptide fermentation, and related fermentation technologies for each region are introduced. The raw materials, microorganisms involved, and usage of ethnic fermented foods are reviewed and compared one another. The beginning of food fermentation technology is related to the invention and use of earthenware, and Northeast Asia is presumed to be one of the birth places. During the period of primitive pottery culture (8000–3000 BCE) on the Korean Peninsula and the coastal region of Korea Strait, boiling technology of food materials using pottery and salt manufacturing technology from seawater were developed, and at the same time, alcohol fermentation using grains and salt-fermentation of fish and vegetables emerged. In the West, where people were nomads, a roasting/grilling culture was continued for a long time, and technologies for fermenting fruit wine and milk products such as cheese and yogurt were developed. As a result, fermentation was mainly used for enhancing the taste of plant foods in the East, while for extending the shelf life of animal foods in the West. The production of salty and meaty flavors from soybeans and marine products by fermentation in East Asia is a technology that increases the value of low-quality proteins.

High-throughput proliferation and activation of NK-92MI cell spheroids via a homemade one-step closed bioreactor in pseudostatic cultures for immunocellular therapy.

The NK-92MI cell line has displayed significant promise in clinical trials for cancer treatment. However, challenges persist in obtaining sufficient cell quantities and achieving optimal cytotoxicity. The proliferation of natural killer (NK) cells involves the formation of cell aggregates, but excessively large aggregates can impede nutrient and waste transport, leading to reduced cell survival rates. In this study, a custom bioreactor was designed to mimic pseudostatic culture conditions by integrating brief mechanical rotation during a 6-h static culture period. This method aimed to achieve an optimal aggregate size while improving cell viability. The findings revealed a 144-fold expansion of 3D NK-92MI cell aggregates, reaching an ideal size of 80-150µm, significantly increasing both cell proliferation and survival rates. After 14days of culture, the NK-92MI cells maintained their phenotype during the subsequent phase of cell activation. Moreover, these cells presented elevated levels of IFN-γ expression after IL-18 activation, resulting in enhanced NK cell-mediated cytotoxicity against K562 cells. This innovative strategy, which uses a closed suspension-based culture system, presents a promising approach for improving cell expansion and activation techniques in immunocellular therapy.

Diachronic evolution in subsistence strategies of ancient humans from the Neolithic Age to the Bronze Age in the Western Liaohe River Basin and its influencing factors

The Western Liaohe River Basin, rich in prehistoric cultures, is one of the significant origins of Chinese civilization. However, what is the subsistence basis that supports the development of prehistoric cultures in this region? More importantly, which role did agriculture play in the development of various prehistoric cultures? Which factors drive the diachronic evolution in subsistence strategies? These issues are worthy of study. Based on this, we systematically collected zooarchaeological, archaeobotanical, and stable isotope data from various prehistoric archeological cultural sites spanning from the Neolithic Age to the Bronze Age in the Western Liaohe River Basin, attempting to explore their economic characteristics, evolution process as well as driving factor. The study found that in the Western Liaohe River Basin, the cultivation of millet crops may have begun as early as the Xiaohexi cultural period, and domestic pigs and dogs may have occurred in the Xinglongwa cultural period (Xionglongwa and Xionglonggou site respectively). However, during Hongshan cultural period, collection and hunting (cervids) remained the main subsistence strategy. In the Lower Xiajiadian cultural period, millet agriculture and domestic pig breeding replaced gathering and hunting, as the predominant subsistence strategy. This subsistence pattern was also adopted by subsequent Upper Xiajiadian cultural populations, and in some areas, animal husbandry developed during this period. We analyze that the evolution of the subsistence strategies of the prehistoric humans in this region may be related with Holocene climate change. Meanwhile, other factors such as improvements in production technology, population pressure and the environmental impacts they brought may also play a role. This study reveals the evolution process of subsistence strategies of prehistoric cultures in the Western Liaohe River Basin, and is of great significance in understanding the relationship between subsistence evolution of ancient populations and environmental changes in northern China.

Analyzing the impact of climatic conditions on rainfed wheat yield in northwest Iran: a parametric and nonparametric approach

ABSTRACT It is essential to comprehend the relationship between agricultural yields and climatic conditions, especially concerning food security and the possible threats to crop output. Wheat is a crucial agricultural crop that covers a significant amount of rainfed production regions in Iran. This study utilized parametric and nonparametric approaches to assess rainfed wheat yield. The study centered on the Tabriz area in northwest Iran, examining precipitation patterns concerning rainfed wheat cultivation. The study focused on analyzing seasonal and distinct rainfall patterns during the cultivation period, utilizing widely recognized drought metrics such as the standardized precipitation index and the standardized precipitation–evaporation index. The study findings indicate a notable upward trend in rainfed wheat output over the analyzed period. The Mann–Kendall test resulted in a p-value of 0.031, indicating statistical significance for the observed rising trend. We conducted trend removal and normalized rainfed wheat yield figures based on seasonal precipitation to study the data more thoroughly. The second phase of the growing season was particularly notable, spanning from the completion of germination to the beginning of blooming. Instead of analyzing precipitation for the full growing season, concentrating on autumn precipitation or the time from germination to blooming might improve yield forecasts and determinations.

Predicting Green Water Footprint of Sugarcane Crop Using Multi-Source Data-Based and Hybrid Machine Learning Algorithms in White Nile State, Sudan

Water scarcity and climate change present substantial obstacles for Sudan, resulting in extensive migration. This study seeks to evaluate the effectiveness of machine learning models in forecasting the green water footprint (GWFP) of sugarcane in the context of climate change. By analyzing various input variables such as climatic conditions, agricultural data, and remote sensing metrics, the research investigates their effects on the sugarcane cultivation period from 2001 to 2020. A total of seven models, including random forest (RF), extreme gradient boosting (XGBoost), and support vector regressor (SVR), in addition to hybrid combinations like RF-XGB, RF-SVR, XGB-SVR, and RF-XGB-SVR, were applied across five scenarios (Sc) which includes different combinations of variables used in the study. The most significant mean bias error (MBE) was recorded in RF with Sc3 (remote sensing parameters), at 5.14 m3 ton−1, followed closely by RF-SVR at 5.05 m3 ton−1, while the minimum MBE was 0.03 m3 ton−1 in RF-SVR with Sc1 (all parameters). SVR exhibited the highest R2 values throughout all scenarios. Notably, the R2 values for dual hybrid models surpassed those of triple hybrid models. The highest Nash–Sutcliffe efficiency (NSE) value of 0.98 was noted in Sc2 (climatic parameters) and XGB-SVR, whereas the lowest NSE of 0.09 was linked to SVR in Sc3. The root mean square error (RMSE) varied across different ML models and scenarios, with Sc3 displaying the weakest performance regarding remote sensing parameters (EVI, NDVI, SAVI, and NDWI). Effective precipitation exerted the most considerable influence on GWFP, contributing 81.67%, followed by relative humidity (RH) at 7.5% and Tmax at 5.24%. The study concludes that individual models were as proficient as, or occasionally surpassed, double and triple hybrid models in predicting GWFP for sugarcane. Moreover, remote sensing indices demonstrated minimal positive influence on GWFP prediction, with Sc3 producing the lowest statistical outcomes across all models. Consequently, the study advocates for the use of hybrid models to mitigate the error term in the prediction of sugarcane GWFP.

Study on the prehistoric decorated spindle whorls in Haidai region of China

Spindle whorls are a kind of spinning tools of crucially important in prehistoric period. Haidai region is one of the main historical and cultural areas of prehistoric China, and had a relatively developed textile handicraft production, where a large number of spindle whorls were unearthed, including many decorated ones. In order to make clear the uses of the decorated spindle whorls and the meanings of the patterns on them, the data of prehistoric decorated spindle whorls unearthed in archaeological sites of the period from Beixin Culture to Yueshi Culture in Haidai region of China were collected and analyzed, and the morphological data (dimension and shapes) of them were compared with that of undecorated ones. The patterns on the decorated spindle whorls were analyzed and classified, and the characteristics of the patterns in different cultural periods were studied, and based on this, the purpose of making decorated spindle whorls and the function and meanings of the patterns were discussed. The results showed that the range of morphological data of the decorated spindle whorls in Haidai region is consistent with that of undecorated ones belonging to the same period and unearthed in the same site, while compared with undecorated spindle whorls, decorated ones are more affected by regional culture and thus show different features from the ones unearthed in other archaeological sites belonging to the same cultural period. It is concluded that the most basic function of the decorated spindle whorls should be as practical tools for normal spinning production, and the patterns on them should have multiple meanings such as practical, decorative, distinctive and symbolic. The study of decorated spindle whorls can help us understand the prehistoric regional culture in Haidai region.

Exploring newly isolated thermotolerant, halotolerant and antimicrobial resistant Bacillus subtilis ProNTL1 from tannery waste and its alkaline protease production.

A hydrolytic enzyme called protease from microbial sources has expansive applications in leather as well as tannery industries and thus leaves no choice but to discover a potential candidate for efficient yield of protease with eccentric characteristics. In the current study, using skim milk agar medium, three bacterial strains (Pro SS14, Pro NTL1, Pro SM) were identified for protease production out of thirty tannery waste samples. The bacterial isolates were identified through morphological, biochemical and molecular basis and the highest protease producer (134U/mL) having the paramount protein content (8.75%) with the greatest biomass (absorbance 1.7 at 600nm) was further bioinformatically identified as Bacillus subtilis Pro NTL1. Moreover, the cultural conditions-pH, temperature, incubation period and salinity were tested to get the optimized condition for the three isolates where Bacillus subtilis Pro NTL1 gave best growth at pH 9 demonstrating alkaline protease producing capability, at 45°C for 72h proving its thermotolerance characteristics and at 3% NaCl concentration declaring its halotolerance property. Furthermore, following Kirby-Bauer agar disc diffusion technique, the antimicrobialsusceptibility pattern was investigated showing Bacillus subtilis Pro NTL1 was resistant to ampicillin, penicillin, sulfafurazole and sensitive to amoxicillin, gentamycin, cefotaxime, kanamycin, imipenem, tetracycline. Additionally, Bacillus subtilis Pro NTL1 showed antagonistic activity against three pathogens named Bacillus cereus, Bacillus megaterium and Escherichia coli. Thus, these findings suggest that Bacillus subtilis Pro NTL1 is a newly isolated strain with unique features which might help to contribute in several biotechnological purposes with significant industrial benefits.

Optimal incubation duration of liquid cultures for assessing culture negative conversion in patients with Mycobacterium avium complex and Mycobacterium abscessus pulmonary diseases.

Mycobacterial liquid culturing typically requires six weeks or longer, primarily because of the slow growth rate of Mycobacterium tuberculosis. This study aimed to evaluate the potential of shortening the duration of mycobacterial liquid culturing in healthcare settings with high prevalence rates of non-tuberculous mycobacteria. We retrospectively analyzed the relationship between mycobacterial species and time to positive testing of liquid cultures from sputum samples using the Mycobacteria Growth Indicator Tube system over a 3.5-year period beginning in July 2020 at a university hospital in Japan. We analyzed 15,147 sputum culture samples and found a 1.1% positivity rate for Mycobacterium tuberculosis complex, while the rates for Mycobacterium avium complex and Mycobacterium abscessus were 17.6% and 2.1%, respectively. The median time to positivity was 17 days for Mycobacterium tuberculosis complex, 9 days for Mycobacterium avium complex, and 4 days for Mycobacterium abscessus. Comparing a 4-week culture period with an eight-week period, the positivity rates for Mycobacterium avium complex and Mycobacterium abscessus were 97.0% and 99.4%, respectively. In settings with a high incidence of non-tuberculous mycobacteria, the basic liquid culturing period can be safely shortened to 4 weeks without significantly compromising detection sensitivity, except for the samples that are highly suspected to contain tuberculosis, extremely slow-growing mycobacteria, smear-positive, or nucleic acid amplification testing positive.

Compatibility of dry incubator on in vitro production of bovine embryos

Embryo culture is crucial to achieve successful outcomes in in vitro production-embryo transfer for cattle. This study explored the innovative use of dry incubators for bovine embryo culture, building on their advantages in human medicine, such as reduced contamination risk, stable temperature control, and lower gas consumption. In this study, we examined changes in osmotic pressure, the in vitro developmental potential of IVP embryos including the cleavage rate, blastocyst development rate, blastocyst diameter, and blastocyst cell number, morphokinetics, and the transcriptional profile of the blastocysts between humidified and dry incubators. Our research demonstrates the feasibility of this approach, showing that although the osmotic pressure gradually increases over the culture period (on day 8: 271.7 vs. 299.0, respectively; P < 0.09), it did not negatively affect the blastocyst formation rate (62.4% vs. 69.8%) and the morphological quality of blastocysts (diameter: 237.4 vs. 242.8, total cell number: 189.2 vs. 242.8). Embryos cultured in dry incubators exhibited morphokinetics comparable to those cultured in conventional humidified incubators. Furthermore, RNA-seq revealed that while a few genes showed changes, the transcriptomic profiles of blastocysts cultured in dry incubators were largely similar to those of blastocysts cultured in humidified incubators. These findings highlight the considerable potential of dry incubators for the in vitro production of bovine embryos.

Nutritional Values of Foxtail Millet (Setaria italica) and other Millets Used for Common Diseases and Management

Abstract: The ever-increasing population and malnutrition in tropical and sub-tropical countries upsurge the demand and security for poor people’s food and nutrition. In erratic climatic conditions, millets can survive as they require less water, short cultivation period and are somewhat tolerant to biotic (bacteria and fungi) and biotic stresses (drought, salinity, nutrient deficiency, etc). Millets possess several nutrient rich components and several health benefits. Despite having superior nutritional, health and agricultural advantages, millets have not received their due attention. Among the millets, foxtail millet is one of the major millets in terms of its second in global production and millets’ yield, quality, and quantity are affected by various diseases. This review provides an overview of the origin and distribution of eight millets, their morphological characteristics with chemical composition, potential health benefits and the fungal, bacterial and viral diseases affecting the growth and yield of millets and their effective management in the framework of ensuring food and nutritional securities in the tropical and sub-tropical countries.

Effects of functional microbial agents on the microbial community and fertility of reclaimed soil in a coal mining area

Low soil fertility is the main factor limiting ecological restoration in coal mining reclamation areas. To improve the nutrient status and microbial community structure of reclaimed soil in coal mining areas, we tested a management strategy combining functional microbial agents, NPK, organic fertilizer, and straw to amend reclaimed soil. We selected corn as the crop for our greenhouse pot experiment, with a cultivation period of 80 days. Greenhouse pot trials were conducted with six treatments: no microbial agent (NMA), including CK (NPK), M (organic fertilizer and NPK), and S (straw and NPK); microbial agent (MA), including B (microbial and NPK), BM (microbial agent, organic fertilizer and NPK fertilizer), and BS (microbial agent, straw and NPK fertilizer). The results showed that the adding microbial agents changed the community structure of bacteria and fungi, and significantly changed the indicator microbial composition. The MA treatment group had an increase in five bacterial microbial network modules related to C and N functionalities such as Proteobacteria and Nitrospirae. In comparison to the B treatment, the combination of microbial agents with organic materials (BM and BS) significantly enhanced soil nutrient contents, including soil organic carbon (SOC), total nitrogen (TN). Therefore, the addition of functional microbial agents (Sphingomonas leidyi, Paenibacillus amylolyticus, and Paenibacillus mucilaginosus), especially in combination with organic materials, can significantly improve the contents of carbon, nitrogen and phosphorus nutrients, bacterial functional activity, and beneficial bacterial abundance in reclaimed soil, which is essential for the rapid improvement of reclaimed soil fertility in coal mining areas.

Astragaloside IV Protects Against IL-1β-Induced Chondrocyte Damage via Activating Autophagy.

Osteoarthritis (OA) is a chronic inflammatory condition that affects the articular cartilage. Astragaloside IV (AS-IV) constitutes the primary active component of the Chinese herbal medicine Huangqi (Radix Astragali Mongolici). AS-IV demonstrates anti-inflammatory and anti-apoptotic attributes, exhibiting therapeutic potential across various inflammatory and apoptosis-related disorders. Nevertheless, its pharmaceutical effects in OA are yet to be fully defined. This study aimed to investigate the protective impact of AS-IV on rat chondrocytes treated with IL-1β and ascertain whether autophagy plays a role in this effect. Chondrocytes were isolated and cultivated from the knee joints of neonatal SD mice. The study included the blank control group, the model group, and the AS-IV concentration gradient group (50, 100, 200 μmol/L) to intervene with chondrocytes. The MTT assay was employed to assess cell viability at varying culture periods, enabling the determination of suitable concentration and duration. Subsequently, chondrocytes were treated with the optimal AS-IV concentration and divided into three groups: the model group replicated IL-1β-induced inflammatory chondrocyte injury, the AS-IV group received a co-culture of AS-IV and IL-1β, and a blank control group was established. Changes in cell morphology and structure were observed using ghost pen cyclic peptide staining. ELISA was used to measure TNF-α and GAG levels in cell supernatants. RT-qPCR assessed p62 and LC3 mRNA expression, while Western Blot evaluated p62 and LC3Ⅱ/Ⅰ protein expression. AS-IV promoted chondrocyte proliferation and concurrently inhibited cell apoptosis. An optimal AS-IV dose of 200 μmol/L and a suitable reaction time of 48 h were identified. Ghost pen cyclic peptide staining indicated that the model group's cytoskeleton exhibited fusiform changes with reduced immunofluorescence intensity, as opposed to the blank control group. The AS-IV group displayed more polygonal cytoskeletal morphology with increased immunofluorescence intensity. AS-IV reduced TNF-α levels and elevated GAG levels in the culture supernatant. Additionally, AS-IV lowered p62 mRNA and protein expression while increasing LC3 mRNA expression in cultured chondrocytes. Our findings suggest that AS-IV mitigates inflammatory chondrocyte injury, safeguarding chondrocytes through a potential autophagy suppression mechanism. These results imply that AS-IV could offer preventive advantages for OA.

Microfluidic device-assisted 3D cell spheroids isolation, staining and embedding

3D cell spheroids have emerged as indispensable tools in cell biology research. In this study, the efficacy of an open-close switchable microfluidic device for on-chip staining and agar-embedding of 3D spheroids was thoroughly validated, especially in preparation for immunohistochemistry (IHC) analysis. The on-chip staining procedure, employing Trypan blue and Calcein-AM/PI staining, was demonstrated to evaluate the cell viability of spheroids. During overextended culture periods, a discernible decline in cell viability was observed at the central region of the spheroids, offering valuable information on the temporal aspects of spheroid development. On-chip agar embedding stands out for its precise control over the location and orientation of three-dimensional spheroids, a key factor in exploring cellular architecture, especially in fused spheroids using IHC analysis. In comparison to tube-based staining, this method significantly cuts down on reagent use and analysis time. This not only boosts efficiency but also lowers the risk of spheroid damage or loss during staining, highlighting its potential for handling miniaturized tissue samples and biopsies in cell assays. The on-chip staining and agar-embedding techniques presented here not only provide valuable insights into spheroids’ behavior but also pave the way for further developments in miniaturized tissue analysis and cell-based assays.

중앙아시아 불교유적의 분포현황과 의의

Buddhist remains in Central Asia are found in five regions: Bactria-Toharistan region in southern Uzbekistan and in southwestern Tajikistan, Fergana region in eastern Uzbekistan and Sogdiana region in central Samarkand, Merv region in eastern Turkmenistan, and Semirechiye region in north-eastern Kyrgyzstan. They are dated from around the 1st century to the 9th and 10th centuries. Most of the Buddhist remains in the northern part of Bactria-Toharistan region are Buddhist temples, but there are only stupas remained without temple buildings such as the Jurmala Stupa and Airtam Stupa. The Karatepa Buddhist Temple, the Fayaztepa Buddhist Temple, and the Buddhist Temples inside and outside the capital city Dalverzintepa are built on the Kushan period in the 2nd and 4th centuries and stupas are found in all these temples. Semirechiye is the region where Buddhist remains are clustered following Bactria-Toharistan region and was influenced by Chinese Buddhism after the 7th century and Tibetan Buddhism later. Not stupas but a large Buddha statue and a large statue of nirvana are found at Akbeshim Buddhist Temple and Ajinatepa Buddhist Temple, respectively, that represents the change in the object of worship. The Buddhist remains of Central Asia are largely divided into two periods. The Bactria-Toharistan Buddhist culture from the 2nd to the 4th centuries was the first period of Buddhist culture in Central Asia, when Buddhism was first introduced and settled in Central Asia along with the expansion of the Kushan Empire. The late Buddhist culture of the Central Asia flourished in Bactria-Toharistan and Semirechiye regions in the 7th and 8th centuries was the second period, and the center of Buddhism moved to East Asia. It was confirmed that Buddhism in Central Asia was developed by exchanging influences with other religions, such as Zoroastrianism previously propagated through traces of rituals related to fire or murals in Buddha statue with a flame-shaped halo. In Buddhist architecture, the use of drying bricks and the construction of stone cave temple that dug into the sandstone layer are reflected in consideration of the material and climate characteristics of the site. As such, it can be seen that the Central Asia did not simply serve as a messenger of civilization, but rather accepted, developed, and spread it to other regions.

Analysis of population heterogeneity in CHO cells by genome-wide DNA methylation analysis and by multi-modal single-cell sequencing

CHO cells are major hosts for the industrial production of therapeutic proteins and their production stability is of considerable economic significance. It is widely known that CHO cells can rapidly acquire genetic alterations, which affects their genetic homogeneity over time. However, the role of non-genetic mechanisms, including epigenetic mechanisms such as DNA methylation, remains poorly understood. We have now used whole-genome bisulfite sequencing to establish single-base methylation maps of eight independent CHO cell lines. Our results identify CpG islands and low-methylated regions as conserved elements with dynamic DNA methylation. Interestingly, methylation patterns were found to cluster clearly along the three main branches of CHO evolution, with no directional changes over short culture periods. Furthermore, multi-ome single-cell sequencing of 9833 nuclei from three independent cultures revealed dynamic subpopulation structures characterized by robust expression differences in pathways related to protein production. Our findings thus provide novel insights into the epigenetic landscape and heterogeneity of CHO cells and support the development of epigenetic biomarkers that trace the emergence of subpopulations in CHO cultures.

Examination of the Effectiveness of Fruit Thinning in the Case of Idaho and Gala Must Early Apple Varieties

Abstract Apple (Malus domestica Borkh.) is the most important fruit in the EU. The total yield is more than 11 million tons.year-1, 15% of which is that of the early apple varieties. Several papers have already been published on the efficacy of crown and fruit thinning in the 20th century. Nowadays, hybrid varieties and intensive crown types are used in modernized systems. Our research was carried out in a Hungarian orchard during two cultivation periods. We studied two early apple varieties (Idaho, Gala Must) to determine how fruit thinning affected the quantity and quality of fruits. We found that the effectiveness of non-thinned technology is similar or slightly better compared to fruit thinning, after the second harvest. The average yield of Idaho apples per tree per harvest was 35.9 vs. 24.8 kg, while that of Gala Must was 50.8 vs. 59.2 kg (with thinning vs. without thinning). To assess quality, we measured water-soluble sugars, NO3 -, NO2 and vitamin C content. Immediately, after the harvest, a surprisingly high vitamin C content was found in the fruits (4.4 mg.100 g-1 on average), which, however, decomposed after 3 months of storage. We also determined the pathway of the decomposition of vitamin C.

ColMA‐based bioprinted 3D scaffold allowed to study tenogenic events in human tendon stem cells

AbstractThe advent of bioprinting has enabled the creation of precise three‐dimensional (3D) cell cultures suitable for biomimetic in vitro models. In this study, we developed a novel protocol for 3D printing methacrylated collagen (ColMa, or PhotoCol®) combined with tendon stem/progenitor cells (hTSPCs) derived from human tendon explants. Although pure ColMa has not previously been proposed as a printable hydrogel, this paper outlines a robust and highly reproducible pipeline for bioprinting this material. Indeed, we successfully fabricated a 3D bioengineered scaffold and cultured it for 21 days under perfusion conditions with medium supplemented with growth/differentiation factor‐5 (GDF‐5). This bioprinting pipeline and the culture conditions created an exceptionally favorable 3D environment, enabling the cells to proliferate, exhibit tenogenic behaviors, and produce a new collagen type I matrix, thereby remodeling the surrounding environment. Indeed, over the 21‐day culture period under perfusion condition, tenomodulin expression showed a significant upregulation on day 7, with a 2.3‐fold increase, compared to days 14 and 21. Collagen type I gene expression was upregulated nearly 10‐fold by day 14. This trend was further confirmed by western blot analysis, which revealed a statistically significant difference in tenomodulin expression between day 21 and both day 7 and day 14. For type I collagen, significant differences were observed between day 0 and day 21, as well as between day 0 and day 14, with a p‐value of 0.01. These results indicate a progressive over‐expression of type I collagen, reflecting cell differentiation towards a proper tenogenic phenotype. Cytokines, such as IL‐8 and IL‐6, levels peaked at 8566 and 7636 pg/mL, respectively, on day 7, before decreasing to 54 and 46 pg/mL by day 21. Overall, the data suggest that the novel ColMa bioprinting protocol effectively provided a conducive environment for the growth and proper differentiation of hTSPCs, showcasing its potential for studying cell behavior and tenogenic differentiation.