Culture Period Research Articles

Enhancement of Forskolin Production Using Aeroponic Cultivation of Coleus forskohlii and the Impact on the Plant Phytochemistry.

Accessing plant resources to extract compounds of interest can sometimes be challenging. To facilitate access and limit the environmental impact, innovative cultivation strategies can be developed. Forskolin is a molecule of high interest, mainly found in the roots of Coleus forskohlii. The aim of this study was to develop aeroponic cultivation methods to provide a local source of Coleus forskohlii and to study the impact of abiotic stress on forskolin and bioactive metabolite production. Three cultivation itineraries (LED lighting, biostimulant, and hydric stress) along with a control itinerary were established. The forskolin content in the plant roots was quantified using HPLC-ELSD, and the results showed that LED treatment proved to be the most promising, increasing root biomass and the total forskolin content recovered at the end of the cultivation period threefold (710.1 ± 21.3 mg vs. 229.9 ± 17.7 mg). Statistical analysis comparing the LED itinerary to the control itinerary identified stress-affected metabolites, showing that LEDs positively influence mainly the concentration of phenolic compounds in the roots and diterpenes in the aerial parts of Coleus forskohlii. Moreover, to better define the phytochemical composition of Coleus forskohlii cultivated in France using aeroponic cultivation, an untargeted metabolomic analysis was conducted using UHPLC-HRMS/MS analysis and molecular networks on both the root and aerial parts. This study demonstrates that aeroponic cultivation, especially with the application of an LED treatment, could be a very promising alternative for a local source of Coleus forskohlii leading to easy access to the roots and aerial parts rich in forskolin and other bioactive compounds.

Induction and Suspension Culture of Panax japonicus Callus Tissue for the Production of Secondary Metabolic Active Substances.

Using Panax japonicus as research material, callus induction and culture were carried out, and high-yielding cell lines were screened to establish a suspension culture system that promotes callus growth and the accumulation of the "total saponins" (total content of triterpenoid glycosides or ginsenosides). Using the root as an explant, the medium for callus induction and proliferation was optimized by adjusting culture conditions (initial inoculation amount, carbon source, shaking speed, hormone concentration, culture time) and a high-yielding cell line with efficient proliferation and high total saponins content was screened out. The conditions of suspension culture were refined to find out the most suitable conditions for the suspension culture of callus, and finally, the suspension culture system was established. We found that the lowest (5%) contamination rate was achieved by disinfecting the fresh roots with 75% alcohol for 60 s, followed by soaking in 10% NaClO for 15 min. The highest induction rate (88.17%) of callus was obtained using the medium MS + 16.11 μmol·L-1 NAA + 13.32 μmol·L-1 6-BA + 30.0 g·L-1 sucrose + 7.5 g·L-1 agar. The callus was loose when the callus subcultured on the proliferation medium (MS + 5.37 μmol·L-1 NAA + 13.32 μmol·L-1 6-BA + 30.0 g·L-1 sucrose + 3.8 g·L-1 gellan gum) for 21 days. The callus growth was cultured in a liquid growth medium (MS + 5.37 μmol·L-1 NAA + 13.32 μmol·L-1 6-BA + 30.0 g·L-1 sucrose) with an initial inoculation amount of 40 g·L-1, a shaking speed of 110 r/min and darkness. Cell growth was fastest with a culture period of 21 days. We replaced the growth medium with the production medium (MS + 5.37 μmol·L-1 NAA + 13.32 μmol·L-1 6-BA + 30.0 g·L-1 glucose) for maximum accumulation of total saponins. [Conclusion] A callus induction and suspension culture system for the root of P. japonicus was established. In this way, we can promote the accumulation of total saponins in callus cells and provide a basis for large-scale cell culture and industrial production of medicinal total saponins.

The roles of cell wall polysaccharides in response to waterlogging stress in Brassica napus L. root

BackgroundBrassica napus L. (B. napus) is susceptible to waterlogging stress during different cultivation periods. Therefore, it is crucial to enhance the resistance to waterlogging stress to achieve a high and stable yield of B. napus.ResultsHere we observed significant differences in the responses of two B. napus varieties in root under waterlogging stress. The sensitive variety (23651) exhibited a more pronounced and rapid reduction in cell wall thickness and root integrity compared with the tolerant variety (Santana) under waterlogging stress. By module clustering analysis based on transcriptome data, we identified that cell wall polysaccharide metabolism responded to waterlogging stress in root. It was found that pectin content was significantly reduced in the sensitive variety compared with the tolerant variety. Furthermore, transcriptome analysis revealed that the expression of two homologous genes encoding polygalacturonase-inhibiting protein 2 (PGIP2), involved in polysaccharide metabolic pathways, was highly upregulated in root of the tolerant variety under waterlogging stress. BnaPGIP2s probably confer waterlogging resistance by inhibiting the activity of polygalacturonases (PGs), which in turn reduces the degradation of the pectin backbone polygalacturonic acid.ConclusionsOur findings demonstrate that cell wall polysaccharides in root plays a vital role in response to the waterlogging stress and provide a theoretical foundation for breeding waterlogging resistance in B. napus varieties.

Variations in Root Characteristics and Cadmium Accumulation of Different Rice Varieties under Dry Cultivation Conditions.

Variations in the cadmium (Cd) accumulation and root characteristics of different genotypes of rice during three developmental periods of dry cultivation were investigated in pot experiments in which two levels of Cd were added to the soil (0 and 10 mg kg-1). The results show that the Cd concentration in each organ of the different rice genotypes decreased in both the order of roots > shoots > grains and during the three developmental periods in the order of the maturity stage > booting stage > tillering stage. The lowest bioaccumulation factor (BCF) and translocation factor (TF) were found in Yunjing37 (YJ37) under Cd stress. At maturity, Cd stress inhibited the root length of Dianheyou34 (DHY34) the most and that of Dianheyou 918 (DHY918) the least, also affecting the root volume of DHY34 and Dianheyou615 (DHY615) the most and that of YJ37 and Yiyou 673 (YY673) the least; the inhibition rates were 41.80, 5.09, 40.95, and 10.51%, respectively. The exodermis showed the greatest thickening in YY673 and the lowest thickening in DHY615, while the endodermis showed the opposite result. The rates of change were 16.48, 2.45, 5.10, and 8.49%, respectively. The stele diameter of DHY615 decreased the most, and that of YY673 decreased the least, while the secondary xylem area showed the opposite result; the rates of change were -21.50, -14.29, -5.86, and -26.35%, respectively. Under Cd stress treatment at maturity, iron plaque was extracted using the dithionite-citrate-bicarbonate (DCB) method. The concentration of iron (DCB-Fe) was highest in YJ37, and the concentration of cadmium (DCB-Cd) was lowest in DHY34. YJ37 was screened as a low Cd-accumulating variety. The concentration of available Cd in the rhizosphere soil, iron plaque, root morphology, and anatomy affect Cd accumulation in rice with genotypic differences. Our screening of Cd-accumulating rice varieties provides a basis for the dry cultivation of rice in areas with high background values of Cd in order to avoid the health risks of Cd intake.

Bone Marrow and Peripheral Blood Mononuclear Cell Phenotype Changes after Cultivation and Autologous Infusion in Patients with Primary Biliary Cholangitis.

Primary biliary cholangitis (PBC) is a condition affecting the liver and immune system. In this study, the impact of autologous bone marrow-derived mononuclear cell (BM-MNC) transplantation on PBC patients was investigated. Sixteen eligible PBC patients participated at the National Scientific Medical Center in Astana, Kazakhstan, between 2017 and 2022, and BM-MNCs were harvested from their anterior iliac crest. After isolating and cultivating the BM-MNCs, they were infused back into the patient's peripheral veins. Changes in BM-MNC and peripheral blood mononuclear cell (PB-MNC) phenotypes were assessed before and after a 24-hour cultivation period and 72 hours post-transplantation. We monitored liver function parameters over 6-month intervals and conducted flow cytometry analysis to assess CD markers on BM-MNCs before and after cultivation and PB-MNCs before and after transplantation. Statistical analysis included the Friedman test for liver parameters and the Wilcoxon signed-rank test for BM-MNC and PB-MNC comparisons. Our findings revealed significant reductions in liver function tests after multiple transplantations. Flow cytometry analysis before and after a 24-hour culture and autologous BM-MNC infusion revealed the expansion of specific cell populations, with significant increases in CD3+, CD4+, CD16+, CD20+, CD25+, CD34+, CD105+, CD73+, СD117+, and CD34+populations, while CD4+25+, CD34+105+, and CD4+FOXP3+ populations decreased. Interestingly, a contradictory finding was observed with a decrease in bone marrow CD34+105+ cell lines (P=0.03) alongside an increase in peripheral CD34+105+ population (P=0.03). In summary, our study shows that BM-MNC transplantation in PBC patients leads to changes in immune cell populations and liver function. These findings suggest potential therapeutic applications of BM-MNC transplantation in managing PBC and offer insights into the dynamics of immune cells associated with this treatment approach.

Effects of cell culture time and cytokines on migration of dental pulp stem cell-derived chondrogenic cells in collagen hydrogels.

The transplantation of collagen hydrogels encapsulating human dental pulp stem cell (DPSC)-derived chondrogenic cells is potentially a novel approach for the regeneration of degenerated nucleus pulposus (NP) and cartilage. Grafted cell migration allows cells to disperse in the hydrogels and the treated tissue from the grafted location. We previously reported the cell migration in type I and type II hydrogels. It is important to explore further how cell culture time affect the cell motility. In this study, we observed the decreased motility of DPSC-derived chondrogenic cells after culturing for 2 weeks compared with cells cultured for 2 days in these gels. The Alamarblue assay showed the cell proliferation during the two-week cell culture period. The findings suggest that the transitions of cell motility and proliferation during the longer culture time. The result indicates that the early culture stage is an optimal time for cell transplantation. In a degenerated disc, the expression of IL-1β and TNFα increased significantly compared with healthy tissue and therefore may affect grafted cell migration. The incorporation of IL-1β and TNFα into the collagen hydrogels decreased cell motility. The study indicates that the control of IL-1β and TNFα production may help to maintain cell motility after transplantation.

Effect of two different bonding agents on the microleakage used for fluoride releasing pit and fissure sealant.

The evaluation of 5th and 7th generation bonding agents on the extent of micro leakage from sealant such as fluoride in forty extracted human premolar teeth is of interest to dentists. 40 extracted teeth satisfying the inclusion and exclusion criteria were randomly divided into two groups. Group I utilized a bonding agent from the fifth generation, whereas Group II employed a bonding agent from the seventh generation, both treated with Helioseal F Plus sealant. Following a 24-hour period of cultivation at a temperature of 37°C and subjecting the sample to 100 cycles of temperature fluctuation varying in temperature from 5°C to 55°C, the samples were submerged in a 0.2% methylene blue dye mixture for duration of 24 hours and examined under a microscope. Result showed that fifth-generation agent mean microleakage was greater, but not statistically significant. The seventh-generation agent demonstrated minimal microleakage with simpler application.

Study on Enzyme Activity and Metabolomics during Culture of Liquid Spawn of Floccularia luteovirens.

To comprehensively investigate the physiological characteristics and metabolic processes of the mycelium of Floccularia luteovirens (F. luteovirens), a wild edible fungus unique to the plateau region, we conducted an in-depth analysis of the mycelium enzyme activity and metabolites during different culture periods. The activity of seven enzymes all followed a trend of initially increasing and then decreasing. The intra- and extracellular activity peaks of three hydrolases-amylase, protease, and cellulase-all occurred on the 20th day, except for the extracellular amylase, which peaked on the 15th day. In contrast, the peak activity of laccase occurred on the 10th day. Moreover, three types of oxidoreductases in the mycelium (catalase (CAT), superoxide dismutase (SOD), and 2,3,5-triphenyltetrazolium chloride (TTC)-dehydrogenase (TTC-DH)) also exhibited significant changes in activity. CAT and SOD activity reached their maximum on the 20th day, whereas TTC-DH showed high activity on both the 10th and 20th days. Through a comprehensive assessment of the evolving trends of these physiological parameters, we determined that the optimal cultivation cycle for F. luteovirens liquid spawn is 20 days. An untargeted metabolomic analysis revealed that 3569 metabolites were detected in the F. luteovirens mycelium, including a variety of secondary metabolites and functional components, with terpenoids being particularly abundant, accounting for 148 types. By comparing three different culture stages (10 days, 20 days, and 30 days), 299, 291, and 381 metabolites, respectively, showed different accumulation patterns in the comparison groups of 10d vs. 20d, 20d vs. 30d, and 10d vs. 30d. These differential metabolites were primarily concentrated in carboxylic acids and their derivatives, fatty acyl groups, organic oxygen compounds, and lipid compounds. In addition, there were several amino acids whose abundance continued to grow during culturing. The metabolism of amino acids greatly affects mycelium growth and development. This research delineates the interplay between mycelium growth and metabolism, offering empirical support for a cultivation strategy for liquid F. luteovirens, and an exploration of its metabolites for potential applications.

The Selection of Lettuce Seedlings for Transplanting in a Plant Factory by a Non-Destructive Estimation of Leaf Area and Fresh Weight

Selecting uniform and healthy seedlings is important to ensure that a certain level of production can be reliably achieved in a plant factory. The objectives of this study were to investigate the potential of non-destructive image analysis for predicting the leaf area and shoot fresh weight of lettuce and to determine the feasibility of using a simple image analysis to select robust seedlings that can produce a uniform and dependable yield of lettuce in a plant factory. To vary the range of the leaf area and shoot fresh weight of lettuce seedlings, we applied two- and three-day irrigation intervals during the period of seedling production and calculated the projected canopy size (PCS) from the top-view images of the lettuce seedlings, although there were no significant growth differences between the irrigation regimes. A high correlation was identified between the PCS and shoot fresh weight for the lettuce seedlings during the period of seedling production, with a coefficient of determination exceeding 0.8. Therefore, the lettuce seedlings were classified into four grades (A–D) based on their PCS values calculated at transplanting. In the early stages of cultivation after transplanting, there were differences in the lettuce growth among the four grades; however, at the harvest (28 days after transplanting), there was no significant difference in the lettuce yield between grades A–C, with the exception of grade D. The lettuce seedlings in grades A–C exhibited the anticipated yield (150 g/plant) at the harvest time. In the correlation between the PCS and leaf area or the shoot fresh weight of lettuce during the cultivation period after transplanting and the entire cultivation period, the R2 values were higher than 0.9, confirming that PCS can be used to predict lettuce growth with greater accuracy. In conclusion, we demonstrated that the PCS calculation from the top-view images, a straightforward image analysis technique, can be employed to non-destructively and accurately predict lettuce leaf area and shoot fresh weight, and the seedlings with the potential to yield above a certain level after transplanting can be objectively and accurately selected based on PCS.

Perturbations to common gardens of anaerobic co-digesters reveal relationships between functional resilience and microbial community composition.

We report the relationship between enrichment of adapted populations and enhancement of community functional resilience in methanogenic bioreactors. Although previous studies have shown the positive effects of acclimation, this work directly investigated the relationships between microbiome dynamics and performance of anaerobic co-digesting reactors in response to different levels of an environmental perturbation (loading of grease interceptor waste [GIW]). Using the methanogenic microbiome from a full-scale digester, we developed eight sets of microbial communities in triplicate using different feed sources. These substrate-specific microbiomes were then exposed to three independent disturbance events of low-, mid- and high-GIW loading rates. This approach allowed us to directly attribute differences in community responses to differences in community composition. Despite identical inocula, environment (digester operation, substrate loading rate, and feeding patterns) and general whole-community function (methane production and effluent quality) during the cultivation period, different substrates led to different microbial community assemblies. Lipid pre-acclimation led to enrichment of a pool of specialized populations, along with thriving of sub-dominant communities. The enrichment of these populations improved functional resilience and process performance when exposed to a low level of lipid-rich perturbation compared with less-acclimated communities. At higher levels of perturbation, the communities were not able to recover methanogenesis, indicating a loading limit to the resilience response. This study extends our current understanding of environmental perturbations, feed-specific adaptation, and functional resilience in methanogenic bioreactors.IMPORTANCEThis study demonstrates, for the first time for GIW co-digestion, how applying similar perturbations to different microbial communities was used to directly identify the causal relationships between microbial community, function, and environment in triplicate anaerobic microbiomes. We evaluated the impact of feed-specific adaptation on methanogenic microbiomes and demonstrated how microbiomes can be influenced to improve their functional (methanogenic) resilience to GIW inhibition. These findings demonstrate how an ecological framework can help improve a biological engineering application, and more specifically, increase the potential of anaerobic co-digestion for converting wastes to energy.

Transcriptional Profiling and Key Enzyme Activity of Epichloë sinensis Isolated from Festuca sinensis in Response to Na2SeO3

Fungal endophytes exhibit symbiotic relationships with their host plants and convert inorganic selenium to organoselenium and Se0. In order to elucidate how Epichloë sinensis from Festuca sinensis adapts to different concentrations of sodium selenate, the dynamic changes of mycelial enzyme activities and metabolic changes at the transcriptional level were documented over a period of 36 h. The activity of enzymes (superoxide dismutase, glutathione reductase, glutathione S-transferase, cysteine synthetase, and methionine synthesis) in mycelia increased in the presence of increased Se concentrations during the cultivation period. The strain with selenium enrichment showed differential changes in gene expression compared to the strain without selenium enrichment, with more changes observed at higher Se concentrations over time. Notably, genes related to ribosomes or ribosome biogenesis in eukaryotes showed significant expression differences among certain groups, with up-regulation of genes involved in oxidoreductase activity, superoxide dismutase, and siderophore biosynthetic processes, and down-regulation of genes involved in steroid biosynthesis. These findings contribute to a better understanding of the transcriptional response of Epichloë sinensis to selenium.

A serum-free culture medium production system by co-culture combining growth factor-secreting cells and l-lactate-assimilating cyanobacteria for sustainable cultured meat production

Large-scale production of cultured meat requires bulk culture medium containing growth-promoting proteins from animal serum. However, animal serum for mammalian cell culture is associated with high costs, ethical concerns, and contamination risks. Owing to its growth factor content, conditioned medium from rat liver epithelial RL34 cells can replace animal serum for myoblast proliferation. More seeded cells and longer culture periods are thought to yield higher growth factor levels, resulting in more effective muscle cell proliferation. However, RL34 cells can deplete nutrients and release harmful metabolites into the culture medium over time, potentially causing growth inhibition and apoptosis. This issue highlights the need for waste clearance during condition medium production. To address this issue, we introduced a lactate permease gene (lldP) and an l-lactate-to-pyruvate conversion enzyme gene (lldD) to generate a recombinant l-lactate-assimilating cyanobacterium Synechococcus sp. KC0110 strain. Transwell co-culture of this strain with RL34 cells exhibited a marked reduction in the levels of harmful metabolites, lactate and ammonium, while maintaining higher concentrations of glucose, pyruvate, and pyruvate-derived amino acids than those seen with RL34 cell monocultures. The co-culture medium supported myoblast proliferation without medium dilution or additional nutrients, which was attributed to the waste clearance and nutrient replenishment effects of the KC0110 strain. This culture system holds potential for the production of low-cost, and animal-free cultured meat.

Effects of Vibrio harveyi infection on the biochemistry, histology and transcriptome in the hepatopancreas of ivory shell (Babylonia areolata)

The ivory shell (Babylonia areolata) is one of the most promising high quality marine products. However, ivory shell is susceptible to Vibrio harveyi infection during the culture period. In this study, we investigated the biochemical indicators, histological changes and transcriptomic response in the hepatopancreas of ivory shells from the PBS control group (PC) and infection group (A3) with 1 × 109 CFU/mL V. harveyi after 24 h. Results showed that compared to the PC group, biochemical indicators, including malondialdehyde (MDA), reactive oxygen species (ROS), acid phosphatase (ACP), and Caspase 3 (Casp-3) were significantly increased (p < 0.05) in A3 group after V. harveyi infection for 24 h. Compared with the PC group, the hepatopancreas of A3 group were seriously damaged, the columnar epithelial cells of the tissue were enlarged, the space of digestive cells was increased, and vacuolar cavities appeared. A total of 95,581 unigenes were obtained and 2949 (1787 up-regulated and 1162 down-regulated) differential expressed genes (DEGs) were identified in the A3 group. GO and KEGG enrichment analysis showed that DEGs were mainly enriched in immune system process (GO:0002376), antioxidant activity (GO:0016209), lysosome (ko04142), toll and IMD signaling pathway (ko04624), and etc. These biological functions and pathways are associated with immune and inflammatory responses and apoptosis. 12 DEGs were randomly selected for real-time quantitative PCR (RT-qPCR) validation, and the expression profiles of these DEGs were consistent with the transcriptome data, confirming the accuracy and reliability of the transcriptome results. In summary, V. harveyi infection of ivory shells inducing oxidative stress, leading to severe hepatopancreatic damage, stimulating glutathione production to neutralize excessive ROS, and stimulating antimicrobial peptides production to counteract the deleterious effects of bacterial infection, which in turn modifying the immune and inflammatory response, ultimately resulting in apoptosis. This study provided valuable information to explore the immune regulation mechanism after V. harveyi infection and established molecular basis to support the prevention of V. harveyi infection.

Effects of Different Tillage Years on Soil Composition and Ground-Dwelling Arthropod Diversity in Gravel-Sand Mulching Watermelon Fields

Arthropods play a crucial role in ecological processes and agricultural productivity. Soil physicochemical properties, indicators of soil health, are closely linked to arthropod communities. Gravel-sand mulching, commonly employed in arid farming, initially enhances water retention and temperature regulation but may contribute to land degradation with prolonged use. This study investigated how varying tillage durations affected soil properties and arthropod diversity under gravel-sand mulching. The analysis employed multiple comparison methods, covariance analysis (ANCOVA), non-metric multidimensional scaling (NMDS), and redundancy analysis (RDA). The results indicated that while soil fertility was better preserved in cultivated fields compared to in the desert grassland, arthropod diversity significantly decreased with longer cultivation periods. A total of 1099 arthropods from 79 species were sampled, by Barber trap. The highest diversity was observed in native grassland (NG), with 305 arthropods from 39 species, while tillage 21 years (GPS-21Y) exhibited the lowest diversity, with only 103 arthropods from 6 species. Dominant species included the carnivores Labidura japonica and Cataglyphis aenes. The analysis revealed low similarity in arthropod communities between GPS-21Y and other fields and high similarity in soil physicochemical properties between NG and the transition zone (STZ). RDA showed available potassium (APP) was negatively correlated with arthropod species diversity and concentration, total Nitrogen (TN) was positively correlated with arthropod species diversity but negatively correlated with species concentration, total phosphorus (TP) was negatively correlated with arthropod species diversity and concentration. This study provides insights into the relationship between maintaining soil fertility and supporting arthropod diversity in grassland agriculture. While soil fertility and arthropod diversity were correlated, continuous cropping practices negatively impacted arthropod diversity, offering valuable information for pest management and sustainable agricultural practices.

Study on the Symbols "Sheng" and "Zu" in Hongshan Culture Rock Paintings

Fertility and ancestor worship are important expressions of social ecology and natural views in Hongshan culture. Based on archaeological findings, numerous archaeological remains related to fertility have been found in Hongshan culture sites. These artifacts reflect the worship, respect, and emphasis on female fertility by the people of Hongshan culture. Additionally, Hongshan culture demonstrates reverence for its ancestors. Archaeological studies have uncovered numerous sacrificial sites and offerings, indicating the reverence and sacrificial activities towards ancestors during the Hongshan culture period. Ancestor worship evolved from primitive totem worship in transitioning from matrilineal to patrilineal societies. Recent surveys and field investigations of rock paintings in the Chifeng region have identified 130 individual "Sheng" and "Zu" character rock paintings, meticulously documented through data recording. Comparative studies of similar "Sheng" and "Zu" character symbols in artifacts and rock paintings from the Hongshan culture period reveal the culture's emphasis on fertility, ancestor worship, and reverence for life. These beliefs and cultural traits were fully expressed in Hongshan culture and profoundly influenced subsequent cultural developments and people's values. This study enhances our understanding of the beliefs and values of the people of Hongshan culture, their regard for life, fertility, and ancestors, and the role of these symbols in cultural transmission.

Improper toy waste handling can harm human health via seafood consumption: A comprehensive health risk assessment of heavy metals

Toy production has been increasing over the last few decades to meet the growing demands for toys across the globe, which has inevitably worsened the problem of toy waste. Given the lack of modern waste disposal facilities, rural villagers in many developing countries often discard and incinerate toy waste in backyards or riverbanks, which may release the pollutants from toys (e.g., heavy metals), contaminate the surrounding areas, and eventually threaten the health of residents. As such, this study examined the impact of improper toy waste handling on the nearby aquaculture site by measuring the contamination level of heavy metals (As, Cd, Cu, Pb, and Zn) in the sediment and seafood (crabs, shrimps, and fish). The health risk of heavy metals via seafood consumption was assessed in different groups of people (males, females, teenagers, and seniors). Results showed that the sediment and seafood at the aquaculture site were generally not contaminated with heavy metals (contamination factor and bioaccumulation factor <1). However, consuming the seafood cultured at this site, especially for crabs, could pose a health risk to humans due to As and Cd (hazard quotient > 1), irrespective of their age and gender. This risk could not be lowered by cooking, except As by boiling. Overall, we revealed that improper toy waste handling did not cause severe heavy metal pollution in the surrounding environment, but the consumption of contaminated seafood could still threaten human health. To safeguard public health, we propose that toy waste should be appropriately treated by the authorities concerned. In addition, consuming seafood with long culture periods (e.g., crabs) should be reduced to minimize the dietary intake of heavy metals and their associated health risk.

Influence of O-GlcNAcylation on KGN cell function

O-GlcNAcylation is a unique form of post-translational glycosylation that affects a variety of cytoplasmic and nuclear proteins of cells. Aberrant O-GlcNAcylation is characteristic of many cancers, and impacts cell proliferation, tumorigenicity and metabolism. O-GlcNAcylation occurs in granulosa cells of ovarian follicles, its expression differs between small (3-5 mm) and large (>8.5 mm) antral follicles, and its manipulation in vitro alters granulosa cell proliferation and metabolism. Here, the aim was to assess whether O-GlcNAcylation similarly occurs in cells from a type of granulosa cell tumor, specifically KGN cells, knowing these cells share functional features of granulosa cells of mature, preovulatory follicles (e.g., FSH-responsiveness and estradiol production). The immortal KGN cell line was utilized to conduct cell culture experiments for the detection and manipulation of O-GlcNAcylation. The cells were grown to confluency in serum containing medium and then sub-cultured in serum-free conditions for immunodetection of O-GlcNAcylation (n = 8 expts.), for cell proliferation (n = 3 expts) and for metabolism assays (n = 12 expts.). The KGN cells were also treated without or with small molecule inhibitors to directly enhance or impair O-GlcNAcylation. Immunoblotting confirmed O-GlcNAc expression in KGN cells, as well as the efficacy of Thiamet-G and OSMI-1 to augment (P < 0.05) and inhibit O-GlcNAcylation (P < 0.05), respectively. Only the inhibition of O-GlcNAcylation compromised KGN cell proliferation (P < 0.05), resulting in a 25 % reduction in proliferation compared to control conditions over a 72 h culture period. Seahorse XFe96 analysis measured effects of O-GlcNAcylation on cellular respiration in the KGN cells. Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) provided indices of glycolysis and oxidative phosphorylation, respectively. During a glycolysis stress test, high glucose increased ECAR and decreased OCR (P < 0.05); oligomycin did not further affect ECAR (P > 0.05), but impaired OCR (P < 0.05); and 2-deoxy-d-glucose decreased ECAR (P < 0.05) without affecting OCR (P > 0.05). Comparatively, a mitochondrial stress test revealed oligomycin increased ECAR (P < 0.05) with a compensatory decrease in OCR (P < 0.05); FCCP increased both ECAR and OCR (P < 0.05); and rotenone + antimycin A decreased both ECAR and OCR (P < 0.05). Manipulation of O-GlcNAcylation in the KGN cells had no effect on ECAR (P > 0.05), but inhibited OCR (P < 0.05). Collectively, the results indicate O-GlcNAcylation occurs in KGN cells, its inhibition impairs cell proliferation, and while KGN cells rely upon both glycolysis and oxidative phosphorylation for cellular respiration, manipulation of O-GlcNAcylation acutely perturbs only oxidative phosphorylation, an effect observed previously with granulosa cells of large antral follicles.

Graft take outcomes of burn patients transplanted with CEA derived from a modified composite culture technique: A case series.

Low-cost and efficient culture environments comparable to standard techniques would undoubtedly improve burn outcomes in under-resourced settings. The aim of this case series was to report on the CEA graft-take using a modified composite culture technique. CEA transplants, following emergency ethical approval, occurred for burn patients (n=25) with low survival prognosis and/or exhausted donor sites. Keratinocytes were retrieved from skin biopsies, seeded centrally on routinely-used dressing gauze and incubated at 37 °C in pediatric incubators. Fresh autogenous plasma was applied daily and hydrogel every third or fourth day. After confluence, the CEA was transplanted onto debrided wound beds. Xenografts were used for temporary cover during the culture period. Final graft take assessment (21 days) was assessed and calculated as a graft take percentage for total CEA area transplanted. Central indices were described as mean (95% CI) and frequency (%) for age, total body surface area, abbreviated burn severity index scores, survival prognosis and graft take. Eleven patients survived with a mean age of 36.1 years (95% CI 25.8-46.4), 45.0 %TBSA burns (95% CI 35.1-54.9), 9.7 ABSI scores (95% CI 8.6-10.8) and 79.5% graft take (95% CI 62.9-96.0). Reduced graft take (61.2%) was observed in patients with perineum burns and increased graft take (97%) in uncomplicated burns. The average CEA graft take was 79.5% using a low-cost culture technique and comparable to the largest case series in literature. The survival of the major burn cases was highly favourable considering injury severity, expected outcomes without CEA and the observed challenges.

Characteristics Of Two Original Facades (Gable and Gunungan) Of Malabero Chinese Shop House In Bengkulu City

Building facades convey a period of culture at the time the building was built, as well as providing the identity of a cultural group or community. Building facades and anthropology became the impetus to find out the original characteristics of the two main facades in the row of Malabero Chinese shophouses on Jalan D. I. Panjaitan Bengkulu City. This research uses the field observation method to obtain data through photographs as well as information from the surrounding community, historians, and cultural experts in Bengkulu and Malabero Chinese Village, to get a link to the facade pattern with an emphasis on building facade elements (roof, walls and building openings) through variables of Chinese shophouse facade characteristics and shop house facade indicators. Malabero Bengkulu's Chinese shophouses are grouped within the Chinese settlement (Chinese Kamp) in front of Fort Marlborough, initially having two characteristics of Chinese shophouse building styles based on their facades. The alteration of the south side of the Malabero Chinese shophouses eliminated many of the original characteristics of the original facade of the Malabero Bengkulu Chinese shophouses (gable roof). As a result, many new characteristics of the shophouse facade elements have been replaced. The shape of the roof (gunungan), the removal of the balcony, and the shape of the building openings became the characteristics of the facade face of the south side Malabero Chinese shop house (gunungan roof). This change in the facade is in anticipation of the load being responsive to earthquakes.

Initiation of Cassava Callus Culture and Its Prospect for Starch Production: A Systematic Mapping Study

Mass propagation of cassava on several hectares of arable land due to increasing demand for its starch is not feasible due to land availability, pests and disease invasion, and long cultivation period. Plant cell culture technology is a promising solution despite the scarcity of cassava callus culture for starch production applications. Therefore, a systematic mapping study (SMS) was performed to identify the applications of cassava tissue culture and its prospects in starch production and investigate the important parameters for cassava callus culture initiation. The SMS began with formulating research questions (RQs), conducting searches on various databases, collecting and screening related articles, and extracting and mapping the selected articles. A total of 56 of 589 articles in the initial searching phase were chosen to be used as references to answer each RQ. The extracted data indicates that cassava tissue culture was mostly used for micropropagation, while starch production from its tissue culture is still limited. Basal medium and plant growth regulators influence cassava callus culture initiation most. The findings of the SMS offer a better understanding of cassava tissue culture and the prospects of producing cassava starch.