Group Culture Research Articles

Medical Grade Honey as a Promising Treatment to Improve Ovarian Tissue Transplantation.

Ovarian tissue cryopreservation is a female fertility preservation technique that presents major challenges for the maintenance of follicular viability after transplantation. The aim of this study was to evaluate and compare the application of L-Mesitran Soft®, a product containing 40% medical grade honey (MGH), with other strategies to improve ovarian grafts’ viability. For this purpose, bovine ovarian tissue was vitrified, warmed and randomly assigned to culture groups: (1) control, (2) MGH 0.2% in vitro, (3) MGH in vivo (direct application in the xenotransplantation), (4) vascular endothelial growth factor (VEGF 50 ng/mL) and (5) vitamin D (100 Nm), during a 48 h period. A sixth group (6) of fragments was thawed on transplantation day and was not cultured. The tissue was xenotransplanted into immunodeficient (Rowett nude homozygous) ovariectomized rats. Grafts were analyzed 48 h after culture, and 7 and 28 days after transplantation. The tissue was subjected to histological and immunohistochemical analysis. Treatments using MGH showed the highest angiogenic and cell proliferation stimulation, with cellular apoptosis, within a healthy cellular turnover pathway. In conclusion, MGH should be considered as a potentially effective and less expensive strategy to improve ovarian tissue transplantation.

Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses

BackgroundTendon healing is clinically challenging largely due to its inferior regenerative capacity. We have previously prepared a soluble, DNA-free, urea-extracted bovine tendon-derived extracellular matrix (tECM) that exhibits strong pro-tenogenic bioactivity on human adipose-derived stem cells (hASCs). In this study, we aimed to elucidate the mechanism of tECM bioactivity via characterization of tECM protein composition and comparison of transcriptomic profiles of hASC cultures treated with tECM versus collagen type I (Col1) as a control ECM component.MethodsThe protein composition of tECM was characterized by SDS-PAGE, hydroxyproline assay, and proteomics analysis. To investigate tECM pro-tenogenic bioactivity and mechanism of action, differentiation of tECM-treated hASC cultures was compared to serum control medium or Col1-treated groups, as assessed via immunofluorescence for tenogenic markers and RNA Sequencing (RNA-Seq).ResultsUrea-extracted tECM yielded consistent protein composition, including collagens (20% w/w) and at least 17 non-collagenous proteins (< 100 kDa) based on MS analysis. Compared to current literature, tECM included key tendon ECM components that are functionally involved in tendon regeneration, as well as those that are involved in similar principal Gene Ontology (GO) functions (ECM-receptor interaction and collagen formation) and signaling pathways (ECM-receptor interaction and focal adhesion). When used as a cell culture supplement, tECM enhanced hASC proliferation and tenogenic differentiation compared to the Col1 and FBS treatment groups based on immunostaining of tenogenesis-associated markers. Furthermore, RNA-Seq analysis revealed a total of 584 genes differentially expressed among the three culture groups. Specifically, Col1-treated hASCs predominantly exhibited expression of genes and pathways related to ECM-associated processes, while tECM-treated hASCs expressed a mixture of ECM- and cell activity-associated processes, which may explain in part the enhanced proliferation and tenogenic differentiation of tECM-treated hASCs.ConclusionsOur findings showed that urea-extracted tECM contained 20% w/w collagens and is significantly enriched with other non-collagenous tendon ECM components. Compared to Col1 treatment, tECM supplementation enhanced hASC proliferation and tenogenic differentiation as well as induced distinct gene expression profiles. These findings provide insights into the potential mechanism of the pro-tenogenic bioactivity of tECM and support the development of future tECM-based approaches for tendon repair.

Hypoxic lung cancer cell-derived exosomal miR-21 mediates macrophage M2 polarization and promotes cancer cell proliferation through targeting IRF1

BackgroundHypoxia is the hallmark of the tumor microenvironment (TME) and plays a critical role during the progress of tumor development. A variety of microRNAs (miRNAs) transmitted by tumor-derived exosomes were involved in intercellular communication. We aimed to elucidate the precise mechanism by which tumor cell-derived exosomes promote lung cancer development by affecting macrophage polarization under hypoxic conditions.MethodsCD163 signal in tumor tissue from lung cancer patients was detected by immunohistochemical (IHC). The M2 polarization-related markers were assessed by flow cytometry and western blot. Exosomes were isolated from normoxic and hypoxic lung cancer cell culture and characterized by transmission electron microscope (TEM), dynamic light scattering (DLS), and western blot. RNA sequencing was performed to show the abnormally expressed miRNAs in exosomes from normoxic and hypoxic lung cancer cell culture. In addition, CCK-8 and clone formation assays were used to assess cell proliferation. Dual luciferase reporter assay was used to evaluate the relationship between miR-21 and IRF1. For in vivo experiment, the male nude mice were injected with H1299 cells with exosomes and miR-21 mimic treatment.ResultsFirstly, we found a strong CD163 signal in tumor tissue from lung cancer patients by IHC. Subsequently, we co-cultured lung cancer cell line H1299 with M0 macrophage THP-1 and found that H1299 in a hypoxic environment promoted THP-1 M2 polarization. PKH67 fluorescence staining experiments confirmed that exosomes of H1299 origin were able to enter THP-1 and induced M2 polarization. RNA sequencing of exosomes showed that miR-21 level was significantly higher in the hypoxic culture group compared to the normoxic group. Subsequent cellular assays showed that miR-21 inhibited the expression of IRF1 by targeting it. In addition, the overexpression of IRF1 reversed the role of miR-21 on macrophage M2 polarization. Finally, we have confirmed through animal experiments that either hypoxic environment or high miR-21 level promoted tumor progression.ConclusionsHigh miR-21 level in hypoxic environments promoted macrophage M2 polarization and induced lung cancer progression through targeting IRF1.

Comparison of pregnancy and live birth rates between fresh day 5 morula transfer and fresh day 6 blastocyst transfer following extended culture for slow growing embryos.

The aim of this study is to evaluate and compare the outcomes of fresh day 5 morula transfers and fresh day 6 blastocyst transfers after extended culture, in women with a whole cohort of slow growing embryos. Fresh embryo transfer cycles of patients that underwent intracytoplasmic sperm injection between 2013 and 2020 with a whole cohort of slow developing embryos on 5thday of fertilization were evaluated for this retrospective cohort study. Outcomes of day 5 morula transfers and day 6 blastocyst transfers after extended culture were compared. Out of 479 patients, day 5 morula transfers were performed to 194 and embryo culturing was extended to day 6 in 285 women. Blastocyst formation was observed in 129 of 285 patients in extended culture group. Implantation rate of day 6 blastocyst transfer group was significantly higher than day 5 morula transfer group (27% vs. 12%, p < 0.001). However, clinical pregnancy (17% vs 15.4%) and live birth rates (14.4% vs 13%) were found similar in day 5 morula transfer and extended culture group per initiated cycle. Although implantation capacity of day 6 blastocysts seems higher comparing to day 5 morulas, clinical pregnancy and live birth rates are similar among intention to treat population in fresh day 5 morula transfers and day 6 blastocyst transfers after extended culture for women that cohorts consist of only slow developing embryos.

Identification of morphometric and intensity of marine leech Zeylanicobdella infestation on cantang grouper from Brondong district, Lamongan and Mandangin Island, Madura

Cantang grouper (Epinephelus fuscoguttatus lanceolatus) is the result of hybridization between female tiger grouper and male grouper fish which is widely cultivated because it has fast growth, easy to breed, and lower cannibalism. Cultivation of cantang grouper can be carried out in semi-intensive ponds and floating net cages (FNC) which have the potential for grouper nursery and rearing activities. One of the problems in the cultivation of cantang grouper is the presence of disease attacks caused by marine leeches Zeylanicobdella with a prevalence of up to 100% in grouper culture in floating net cages. This study aims to determine differences in species and intensity of marine leech Zeylanicobdella infestation in cantang grouper (Epinephelus fuscoguttatus lanceolatus) from semi-intensive ponds in Brondong District, Lamongan and floating net cages (FNC) on Mandangin Island, Madura. The method used in this research is a survey and sampling is done by purposive sampling method. Cantang groupers measuring 15-20 cm from semi-intensive ponds and 20-25 cm from floating net cages each were taken as many as 35 individuals. Identification of Zeylanicobdella marine leeches using a binocular microscope and making sketches of ectoparasites with the help of a binocular microscope equipped with a lucida camera. Marine leech species found to infest the cantang grouper (Epinephelus fuscoguttatus lanceolatus) is Zeylanicobdella arugamensis which was found on the body surface, fins, gills, and eyes. The results of statistical analysis using the Mann-Whitney U test showed that there was no significant difference (p > 0,05) between species and intensity of Zeylanicobdella arugamensis in cantang grouper reared in semi-intensive ponds and floating net cages. The intensity of marine leeches Zeylanicobdella arugamensis in semi-intensive ponds was in the medium category (14,74 individuals/head) and in floating net cages was in the medium category (27,74 individuals/head). The highest intensity of infestation marine leeches Zeylanicobdella arugamensis in floating net cages of 27,74 individuals/head. There was no significant difference in the intensity of Zeylanicobdella arugamensis marine leech infestation in cantang grouper both from semi-intensive ponds and floating net cages (FNC) which were included in the medium category.

Redesigning of 3-Dimensional Vascular-Muscle Structure Using ADSCs/HUVECs Co-Culture and VEGF on Engineered Skeletal Muscle ECM.

Objective The main objective of this study is to determine the myogenic effects of skeletal muscle extracellular matrix, vascular endothelial growth factor and human umbilical vein endothelial cells on adipose-derived stem cells to achieve a 3-dimensional engineered vascular-muscle structure. Materials and MethodsThe present experimental research was designed based on two main groups, i.e. monoculture of adipose tissue-derived stem cells (ADSCs) and co-culture of ADSCs and human umbilical vein endothelial cells (HUVECs) in a ratio of 1:1. Skeletal muscle tissue was isolated, decellularized and its surface was electrospun using polycaprolactone/gelatin parallel nanofibers and then matrix topography was evaluated through H&E, trichrome staining and SEM. The expression of MyHC2 gene and tropomyosin protein were examined through real-time reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence, respectively. Finally, the morphology of mesenchymal and endothelial cells and their relationship with each other and with the engineered scaffold were examined by scanning electron microscopy (SEM).Results According to H&E and Masson’s Trichrome staining, muscle tissue was completely decellularized. SEM showed parallel Polycaprolactone (PCL)/gelatin nanofibers with an average diameter of about 300 nm. The immunofluorescence proved that tropomyosin was positive in the ADSCs monoculture and the ADSCs/HUVECs co- culture in horse serum (HS) and HS/VEGF groups. There was a significant difference in the expression of the MyHC2 gene between the ADSCs and ADSCs/HUVECs culture groups (P<0.05) and between the 2D and 3D models in HS/ VEGF differentiation groups (P<001). Moreover, a significant increase existed between the HS/VEGF group and other groups in terms of endothelial cells growth and proliferation as well as their relationship with differentiated myoblasts (P<0.05). ConclusionCo-culture of ADSCs/HUVECs on the engineered cell-free muscle scaffold and the dual effects of VEGF can lead to formation of a favorable engineered vascular-muscular tissue. These engineered structures can be used as an acceptable tool for tissue implantation in muscle injuries and regeneration, especially in challenging injuries such as volumetric muscle loss, which also require vascular repair.

Eco-Cultural Niche Breadth and Overlap Within the Cucuteni–Trypillia Culture Groups During the Eneolithic

One of the most applied tools for documenting cultural variability and for tracing cultural trajectories within the environmental context is eco-cultural niche modeling and its associated methodology. The niche breadth characterization quantitatively evaluates the links between a given adaptive system and ecological constraints, which provides valuable information for archeology. For this purpose, in this study, 10 independent climatic and topographic environmental variables were interpolated, and eco-cultural niche modeling techniques were used to determine whether these differences in geographic distributions and niche breadth are consequences of differences in five Cucuteni–Trypillia groups that flourished in Eastern Europe during the Eneolithic (cal. 5,400/5,300–2,800/2,700 BCE). Our results showed that the eco-cultural niches of Cucuteni–Trypillia groups are significantly overlapping, and the expansion trend of the last two cultural groups (Late Eneolithic–cal. 4,100/4,000–2,800/2,700 BCE) into the northeastern steppe regions was not due to ecological niche differences but rather a result of other cultural factors. Furthermore, we highlighted that the first three Cucuteni–Trypillia groups (Early-to-Middle Eneolithic—cal. 5,400/5,300–4,100/4,000 BCE) had slightly more constrained ecological niches in the mid-Holocene ecosystems than the Late Eneolithic groups. The results have significant implications for understanding the geographical range dynamics and distribution of the last great Chalcolithic society of Old Europe and contribute to the characterization of ecological niches they have exploited during the cultural evolutionary process.

P-135 Clinical outcomes depending on post-warming culture duration of vitrified-warmed single blastocyst transfer cycles

Abstract Study question What is the impact of post-warming blastocysts after very short-term culture (&amp;lt;2 hours) or short-term culture (2–4 hours) on the clinical outcomes? Summary answer The clinical outcomes of very short-term culture and short-term culture were not significantly different in vitrified-warmed blastocyst transfer. What is known already In vitrified-warmed blastocyst transfer (VBT), complete blastomere survival and mitotic resumption during warming are generally considered to be the most important factors affecting pregnancy outcomes. It has been assumed that a sufficient warming time may be required for resumption of cell proliferation and development. Currently there is evidence that suggests a minimum mean time of 2.70 ± 1.20 necessary to complete the re-expansion of the embryo that allows evaluating its adequate development. However, the effect of culture duration on pregnancy outcomes in VBT remains controversial. Study design, size, duration A retrospective observational study was carried out, with all the patients who had frozen-thawed embryos transferred between January and December 202, in Clínica Alemana, Santiago, Chile. The average standard time for embryo thawing in our center is 2 hours. However, on certain occasions, due to its insufficient development and the pressing schedule of surgery rooms, a second embryo must be thawed, usually in less than 2 hours. Participants/materials, setting, methods Embryos were divided into two study groups depending on whether their post-warming culture period was very short-term (&amp;lt;2 hours) or short-term culture (2–4 hours). Only transfers of 1 embryo on day 5-6 were included. Embryo morphology was analyzed with a Garner microscope score to estimate the quality of the embryos in both groups. Main results and the role of chance During the study period, a total of 127 single VBT cycles were included: 48 in the very short-term group and 79 in the short-term group. Patients’ demographic characteristics did not significantly differ between the groups. (Age 34.9±3.6 vs 35±3.4, p=0.89; Infertility duration (yr) 2.75±1.1 vs 2.5 ±1.1, p=0.24; AMH 2.1 [0.2-5.4] vs 2.1 [0.2-5], p=0.84) An analysis of previous fresh in vitro fertilization cycles and subsequent VBT cycle parameters revealed no differences in the number of retrieved oocytes (11.7 vs 12, p=0.74), the number of fertilized oocytes (8.6 vs 8.5, p=0.86), the number of vitrified blastocysts (3.4 vs 3.6, p=0.87) and in both groups. The clinical pregnancy rate per embryo transferred were similar in the very short-term culture group and the short-term culture group (66.7% vs. 68.4%, p=0.44). There was no significant difference in the ongoing pregnancy rate per embryo transferred between the two groups (18,8% vs. 16.5.%, p=0.74). The miscarriage rate in both groups was comparable (p=0.77). Limitations, reasons for caution The small number of women recruited ant the retrospective design of the study, should be recognized as possible sources of bias. Wider implications of the findings This study showed that the culture time after warming was irrelevant for blastocyst implantation potential, and that the speed of re-expansion was not correlated with clinical outcomes. Embryo transfer timing after warming may be determined by optimizing each laboratory’s work flow. Trial registration number 0

Bacillus subtilis Produces Amino Acids to Stimulate Protein Synthesis in Ruminal Tissue Explants via the Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Beta-Serine/Threonine Kinase-Mammalian Target of Rapamycin Complex 1 Pathway.

BackgroundBacillus subtilis is a probiotic strain that is widely used as a feed supplement for ruminants. In this study, one B. subtilis strain isolated from the ruminal fluid of Holstein dairy cows was used for an ex vivo study with ruminal tissue explants. The main goal was to assess the potential endosymbiotic links between B. subtilis and the ruminal epithelium using molecular analyses and amino acid profiling. The explant culture protocol was first optimized to determine the ideal conditions in terms of tissue viability before performing the actual experiments involving active and inactive bacteria with or without protein synthesis inhibitors, such as LY294002 (phosphatidylinositol 3-kinase inhibitor) or rapamycin [mammalian target of rapamycin (mTOR) inhibitor].ResultsThe mRNA levels of phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta (PIK3CB), serine/threonine kinase (AKT), mTOR, P70S6K1, and eukaryotic translation initiation factor 4E binding protein 1 were the highest (p < 0.01), while those of programmed cell death 4 were the lowest when the tissue was incubated with 107 of B. subtilis. Compared with the inactivated bacteria, the expression levels of PIK3CB and AKT, and overall changes in mTOR and P70S6K1 were greater in rumen explants with living bacteria (p < 0.05). With an increase in B. subtilis concentration, the trends of protein and corresponding gene changes were consistent. There were differences in the concentrations of individual amino acids in the supernatants of living and inactivated bacterial culture groups, with most amino acids enriched in pathways, such as aminoacyl tRNA biosynthesis, cyanoamino acid metabolism, monobactam biosynthesis, or glycine, serine, and threonine metabolism. The addition of psilocybin upregulated the expression levels of PIK3CB and AKT. A significant decrease (p < 0.05) in PIK3CB and mTOR protein expression levels was detected after the addition of LY294002 and rapamycin. In addition, These responses were associated with the downregulation (p < 0.05) of AKT and P70S6K protein expression levels.ConclusionsWe confirmed that the in vivo ruminal tissue culture system is a suitable model for studying probiotic-induced alterations in tissue function. As such, this study provides a means for future mechanistic studies related to microbial regulation and the dietary supply of proteins. In addition, living and inactivated B. subtilis can promote protein synthesis in ruminal tissue explants by altering the expression levels of related factors in the PIK3CB–AKT–mTORC1 pathway, which could further aid in optimizing the feed efficiency and increasing the use of inactivated bacteria as additives in dairy cow farming.

Spermatogenic differentiation of spermatogonial stem cells on three-dimensional silk nanofiber scaffold

BackgroundNano-fibrous scaffolds provide a three-dimensional matrix that guides sufficient orientation of seeded cells similar to a natural niche. In this research, we designed a silk scaffold to improve the differention of mouse spermatogonial stem cells to spermatogenic cell lines. Spermatogonial stem cells were collected from neonatal mouse (2–6 days) testes (n=60) using a two steps mechanical and enzymatic method. Cells were seeded on a silk scaffold and were cultured in Dulbecco’s modified Eagle’s medium, supplemented with 15 % fetal bovine serum and 1000 units/ml leukemia inhibitory factor, and incubated at 32°C in a humidified atmosphere of 5% CO2 in air. SEM technique was done for confirmation of seeding cells.In this study two major groups (i.e., 2D and 3D culture groups) of 30 mice each. Isolated testicular cells from each group were cultured in the absence of silk scaffold or the presence of silk scaffold.For induction of differentiation, seeded cells on a scaffold were exposed to 1 μM and 50 ng/ml BMP-4. The specific spermatogenic genes, e.g.; VASA, DAZL, PLZF, and Piwil2, were assessed via real-time PCR and immunocytochemistry techniques. P values less than 0.05 were assumed significant. All experiments were performed at least three times.ResultsSEM analysis confirmed the homogeneity of fabricated silk scaffold and average diameter of 450 nm for nanofibers fibers. Silk scaffold induces attachment of SSCs in comparison to the monolayer group. Spermatogonia stem cell colonies were observed gradually after 1 week of culture. Electrospun scaffold supports the differentiation of SSCs to spermatogenic lines. Dates of real-time PCR showed that the expression of meiotic markers, VASA, DAZL, and Piwil2 as related to specific spermatogenic genes, had a significant upregulation in cell-seeded silk scaffold compared to the control group (P < 0.05).Immunocytochemistry founding approved the expression of specific spermatogenic markers; DAZL and PLZF were higher in the experiment group compared to the control (P < 0.05).ConclusionIt is concluded silk scaffold induces spermatogenic differentiation of mouse spermatogonial stem cells in vitro.

AACR Cancer Disparities Progress Report 2022.

AACR Cancer Disparities Progress Report 2022.

The different irradiation parameters of carbon dioxide laser effects on periodontal ligament cells

Background/purpose: Photobiostimulation (PBS) can affect cellular functions. The objective of the present study was to evaluate the cellular changes in periodontal ligament (PDL) cells that received different carbon dioxide (CO2) laser irradiation parameters under negative pressure culture. Materials and methodsThe negative pressure-cultured PDL cells on normal medium and differentiation medium were subjected to continuous irradiation with a CO2 laser at an energy density of 5 J/cm2 or 10 J/cm2. The irradiated PDL cells were harvested at Days 1, 5 and 7, and their viability was analyzed by the Presto Blue assay and the biologic markers alkaline phosphatase (ALP), bone sialopoietin (BSP), osteopontin (OPN), osteocalcin (OC), matrix metalloproteinase-3 (MMP-3) collagen I (Col I) and cyclooxygenase-2 (COX-2) expression by reverse transcription-polymerase chain reaction (RT–PCR). ResultsThe PDL cell viability showed that the differentiation medium groups were higher than the normal culture groups. The cell morphologies were all expressed as spindle type. The inflammatory markers in the laser-irradiated groups were higher on the first day and decreased on the seventh day (P < 0.05). Osteogenesis markers were highly expressed at different time periods (P < 0.05). The Col I and OPN genes were highly expressed on the first day, and the Col I high expression lasted until the seventh day. The OC gene was highly expressed on the seventh day. The effects of PDL cultured in differential medium and normal medium were the same in the present study. ConclusionA low-dose CO2 laser continuously irradiating cultured PDL cells can induce osteogenesis and reduce cell inflammatory expression.

Effect of berberine on cancer cell motility in glioma, lung cancer, and prostate cancer cell cultures.

e15079 Background: Initially, berberine was used as an antimicrobial agent in traditional medicine, but its anticancer properties were later discovered. In addition to its cytotoxic effect, berberine also has the ability to inhibit cell motility, which has been demonstrated in some permanent cancer cell lines. The objective of this study was to assess berberine anti-migratory activity in permanent cell cultures compared to primary cell cultures which are generally thought to better reflect tumor characteristics. Methods: H1299 lung cancer, PC3 prostate cancer, and T98G glioma cells, as well as primary cell cultures of the corresponding cancer obtained in our laboratory, were planted in an amount of 15*104 cells per well of a 24-well plate (Biofil, China) in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA). After cell adhesion berberine was added in concentration 5 µM, and a wound healing assay was performed according to the standard procedure. Cell plates were continuously incubated and photographed in Lionheart FX imager (BioTek, USA) at 37°C and 5.0% CO2. The extent of cell migration was measured as the percentage reduction in wound area after 48 hours of incubation relative to baseline. Data are presented as Mean ± 95% confidence interval (n = 12). Results: The use of berberine at a concentration of 5 µM led to a significant decrease in cell motility in permanent cultures of lung cancer, glioma and prostate cancer. Namely, the reduction in the wound area after 48 hours of incubation with bereberine was 74.52±12.3% (compared with 94.56±6.2% in the control) in H1299 cell culture, 38.22±10.6% (compared with 83.89±15.5% in the control) in T98G cell culture, and 48.6±7.5% (compared with 69.56±8.1% in the control) in PC3 cell culture. The resulting difference between the control and experimental groups in permanent cell cultures was statistically significant at a significance level of 5% (df = 22). At the same time, the values of wound area reduction for primary cultures of the same cancers did not differ significantly in the control and under the influence of 5 μM berberine at the accepted level of significance. Namely, the reduction in the wound area after 48 hours of incubation with bereberine was 84.79±11.2% (compared with 81.47±15.3% in the control) in lung cancer primary cell culture, 94.64±5.1% (compared with 91.73±6.8% in the control) in glioma primary culture, and 62.63±5.8% (compared with 61.1±8.9% in the control) in prostate cancer primary cell culture. Conclusions: Permanent cell lines are more sensitive to berberine anti-migratory activity than primary cancer cell lines of the same localization.

Transactional Communication of Garut People in Dealing with Potential Natural Disaster

Villages in West Java are some of the most hydrometeorology disaster-prone areas. Giri Awas in Garut is one of the villages in West Java whose area is located on the hills of mountain slopes. Some areas have undergone transfer from the forest into the plantation, which caused a higher risk of flood and landslides. This research aimed to identify how the communication is carried out by villagers to mitigate the risk of disaster. This research uses the concept of a transactional communication model with a descriptive qualitative method by collecting data through interviews and field observation. The researchers found the transactional communication model as the effort for disaster risk mitigation has been conducted by the village apparatus toward society. However, some aspects of transactional communication have not been optimally done. It has not been conducted continually, and messages related to disaster experiences in the past have not been communicated to the whole society. There are some unused potentials of the people to support the sustainability of transactional communication by utilizing local culture and other social groups. Optimizing society’s potential can support the negotiation of meaning between local government and society.

Scaffold-free 3D culturing enhance pluripotency, immunomodulatory factors, and differentiation potential of Wharton’s jelly-mesenchymal stem cells

Mesenchymal stem cells (MSCs) show a decline in pluripotency and differentiation with increased cell culture passages in 2D cultures. The 2D monolayer culture fails to correctly imitate the architecture and microenvironments of in-vivo cell models. Alternatively, 3D culture may improve the simulations of in-vivo cell microenvironments with wide applications in cell culture and drug discovery. In the present study, we compared various 3D culturing techniques such as 3D micro-well (3D-S), hanging drop (HD), and ultra-low attachment (ULA) plate-based spheroid culture to study their effect on morphology, viability, pluripotency, cell surface markers, immunomodulatory factors, and differentiation capabilities of Wharton’s jelly-mesenchymal stem cells (WJ-MSCs). The cell morphology, viability, and senescence of 3D cultured WJ-MSCs were comparable to cells in 2D culture. The expression of pluripotency markers (OCT4, SOX2, and NANOG) was enhanced upto 2–8 fold in 3D cultured WJ-MSCs when compared to 2D culture. Moreover, the immunomodulatory factors (IDO, IL-10, LIF, ANG1, and VEGF) were significantly elevated in ULA based 3D cultured WJ-MSCs. Furthermore, significant enhancement in the differentiation potential of WJ-MSCs towards adipocyte (ADP and C/EBP-α), osteocyte (OPN and RUNX2), and definitive endodermal (SOX17, FOXA2, and CXCR4) lineages in 3D culture conditions were observed. Additionally, the osteogenic and adipogenic differentiation potential of WJ-MSCs over the time points 7 days, 14 days, and 28 days was also significantly increased in 3D culture groups. Our study demonstrates that stemness properties of WJ-MSCs were significantly enhanced in 3D cultures and ULA-based culture outperformed other methods with high pluripotency gene expression and enhanced differentiation potential. This study indicates the efficacy of 3D cultures to bridge the gap between 2D cell culture and animal models in regenerative medicine.

The microenvironment of silk/gelatin nanofibrous scaffold improves proliferation and differentiation of Wharton’s jelly-derived mesenchymal cells into islet-like cells

The use of umbilical cord-derived mesenchymal stem cells along with three-dimensional (3D) scaffolds in pancreatic tissue engineering can be considered as a treatment for diabetes. This study aimed to investigate the differentiation of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) into pancreatic islet-insulin producing cells (IPCs) on silk/gelatin nanofibers as a 3D scaffold. Mesenchymal markers were evaluated at the mesenchymal stem cells (MSCs) level by flow cytometry. WJ-MSCs were then cultured on 3D scaffolds and treated with a differential medium. Immunocytochemical assays showed efficient differentiation of WJ-MSCs into IPCs. Also, Real-time PCR results showed a significant increase in the expression of pancreatic genes in the 3D culture group compared to the two-dimensional (2D) culture group. Despite these cases, the secretion of insulin and C-peptide in response to different concentrations of glucose in the 3D group was significantly higher than in the 2D culture. The results of our study showed that silk/gelatin scaffold with WJ-MSCs could be a good option in the production of IPCs in regenerative medicine and pancreatic tissue engineering.

Technological Traditions in Pottery of the Kulay Cultural-Historical Community on Barsova Gora (Based on the Materials of the Sites Barsov Gorodok I/5 and I/7)

Purpose. Barsova Gora is a unique archaeological and landscape site located in the Tyumen Region of the Russian Federation. The archaeological sites Barsov gorodok I/5 and I/7 are located on the edge of the bank of Utoplaya River. The aim of the study is to reconstruct the content of the production steps in pottery of the surgut variant of the kulai culture in the sites Barsov gorodok I/5 and I/7. Results. A technical and technological analysis was carried out for 33 vessels from the site Barsov gorodok I/5 and for 5 vessels from the site Barsov gorodok I/7. Nine recipes of clay paste have been found at the site Barsov gorodok I/5. The main ones are unmixed recipes Clay + Broken stone, Clay + Chamotte. Recipes with the addition of sand, introduced in a ratio of 1 : 1, stand out. Three recipes have been found on the site Barsov gorodok I/7. Vessels were molded with the help of flaps. Cases of using tapes with an overlay have been recorded. Various combinations of tools for surface treatment. 22 variants have been found on the site Barsov gorodok I/5. On the site Barsov gorodok I/5, 4 variants were found for 5 vessels. On both sites, a specific method of smoothing down with a serrated tool was recorded on the inside of the transition point from the shoulder to the body. Ware could be fired in a reducing and reducingoxidizing environment. Conclusion. The pottery skills of the population of both settlements are similar to the general pottery traditions of the kulay culture on Barsova Gora. This is manifested in the similarity of the clays used, a close range of artificial impurities and recipes for clay paste, variability in surface treatment, and the presence of specific technological skills. The difference is manifested in the skills of constructing a hollow body and in the use of sand as an artificial impurity. The use of patchwork molding along with the band molding reflects the coexistence of two groups of potters with different skills in vessel construction at these sites. This may be due to the interaction within these settlements of populations with different technological traditions or to the influence of earlier pottery traditions of previous cultures. The addition of sand may be related to the interaction of this population with the more eastern groups of the kulay culture, for which this admixture is dominant, or be the result of mixing with an earlier pottery tradition.

Yeast culture (Saccharomyces cerevisiae) and its active metabolites affect the cecal microbiome of broilers

Yeast cultures (YCs) are defined as promising feed additives that maintain the health of birds and improve growth performance by modulating gut microbiota. YCs contain effective metabolites such as glycine, fructose, inositol, galactose, and sucrose. This study investigated the effects of YCs and their effective metabolites on carcass traits and cecal microflora in broilers. A total of 280 one-day-old mixed-sex Arbor Acres broilers were randomly allocated to seven groups. The basal diet (control DZ) was supplemented with various proportions of glycine, fructose, inositol, galactose, and sucrose (Groups A, B, and C), 24-hour grown Saccharomyces cerevisiae cultures (Group D) (YC), and a commercial yeast culture product (SZ) at concentrations of 0.1% and 1% (Groups E and F). Bodyweight of broilers was correlated positively with proportions of Proteobacteria in Group C and Lactobacillus and Roseburia in Group B (P &lt;0.05). Broilers fed diets supplemented with YC or its active metabolites had the highest proportions of bacteria involved in nucleotide metabolism, and amino acid and carbohydrate metabolism. These results suggested that the dietary addition of YC could alter the proliferation of beneficial bacteria in broilers.

MP05-10 MIXING IT UP: ARE PATIENTS WITH MIXED URINE CULTURES AT HIGHER RISK OF POSTOPERATIVE SEPSIS AFTER URETEROSCOPY?

You have accessJournal of UrologyCME1 May 2022MP05-10 MIXING IT UP: ARE PATIENTS WITH MIXED URINE CULTURES AT HIGHER RISK OF POSTOPERATIVE SEPSIS AFTER URETEROSCOPY? Young Son, Julia Scali, Ian Madison, Benjamin Fink, Katelyn Klimowich, Dayna DeVincentz, Paul Chialastri, and Thomas Mueller Young SonYoung Son More articles by this author , Julia ScaliJulia Scali More articles by this author , Ian MadisonIan Madison More articles by this author , Benjamin FinkBenjamin Fink More articles by this author , Katelyn KlimowichKatelyn Klimowich More articles by this author , Dayna DeVincentzDayna DeVincentz More articles by this author , Paul ChialastriPaul Chialastri More articles by this author , and Thomas MuellerThomas Mueller More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002522.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Ureteroscopy with laser lithotripsy has been used for the treatment of ureteral calculi since the 1980s. Life-threatening urosepsis is one of the most severe complications of this procedure. The American Urological Association (AUA) recommends single-dose antibiotic prophylaxis prior to ureteroscopic stone removal. The significance of colonization in an asymptomatic patient is unknown at this time. Our objective is to determine if preoperative urine cultures with mixed flora on microbiology report, have a higher rate of postoperative sepsis than patients with negative cultures for uncomplicated elective ureteroscopy with laser lithotripsy. METHODS: A retrospective multi-institutional chart review was performed on 306 patients for which procedure code included elective ureteroscopy with lithotripsy from April 2018 until December 2020. Patients were excluded if the mixed urine cultures were treated with antibiotic therapy, inpatient setting, or had percutaneous nephrostomy or nephroureterostomy tubes. Mixed cultures were defined as mixed culture >10,000 CFU (colony forming unit) and positive cultures were defined as bacteria with >100,000 CFU. RESULTS: The overall sepsis rates were 3% with negative culture preoperatively, 10.9% with a mixed culture, and 19.2% with positive culture. There was a significant difference in carbonate apatite composition between mixed culture group and negative culture group (p=0.02) and positive culture group and negative culture group (p=0.01) (Table 1). There was an increased stent duration 38.4 days compared to 16.6 days in septic vs. non-sepsis group (p<0.002). There were discordinate amount of carbonate apatite composition with 37.7% in the sepsis group compared to 14.2% in the non-sepsis group (p=0.003) (Table 2). CONCLUSIONS: We conclude that there is 3.6 times more likely that postoperative sepsis patients had mixed cultures compared to negative cultures. Also, there was 2.7 times more carbonate apatite stone composition in postoperative sepsis group compared to non-sepsis group. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e71 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Young Son More articles by this author Julia Scali More articles by this author Ian Madison More articles by this author Benjamin Fink More articles by this author Katelyn Klimowich More articles by this author Dayna DeVincentz More articles by this author Paul Chialastri More articles by this author Thomas Mueller More articles by this author Expand All Advertisement PDF DownloadLoading ...

Stimulation of autophagy and synaptic maintenance are commonalities induced by an exercise‐mimetic and diet supplement to avoid initiators of age‐related cognitive decline

Brain aging causes synaptic and cellular vulnerabilities, leading to cognitive impairment and an increased risk for dementia. Removal of old and damaged proteins becomes less efficient with age, causing protein accumulation stress and corresponding synaptopathy as protein clearance systems fail. Interestingly, physical exercise and a healthy diet are well known to promote cognitive health, perhaps by delaying or reducing age‐related changes that cause synaptic compromise. It is vital to understand the mechanism by which exercise and diet positively influence brain health since they are the only avenues implicated in reducing dementia risk. Accordingly, we tested if an exercise‐mimetic and a dietary supplement have a common influence on the protein clearing autophagy‐lysosomal pathway, and if such a mechanistic link promotes synaptic maintenance and resilience. Our study used long‐term brain explants that exhibit multi‐neural cell types and subfields, as well as native neuronal circuitries and synaptic density as compared to the intact adult brain. The exercise‐mimetic compound β‐guanidinopropionic acid (β‐GPA), a creatine analog, and a specific extract of American ginseng (P. quinquefolius; USANA Health Sci), were applied daily to the matured, 3‐dimensional cultures of hippocampal tissue for 3‐6 days. The gently harvested explants were then assessed by immunoblot for several markers including the cathepsin B (CatB) component of the autophagy‐lysosomal pathway, a protease found to improve clearance of pathogenic proteins and preserve synaptic and cognitive measures in models of AD, α‐synucleinopathy, and mild cognitive impairment (Mueller‐Steiner et al. 2006 ‐ Neuron 51:703; Butler et al. 2011 ‐ PLoS ONE 6:e20501; Hwang et al. 2019 ‐ Inter J Mol Sci 20:4432). Both β‐GPA and the American ginseng extract improved the levels of the active CatB isoform and increased autophagic flux. In addition, they both had a positive influence on synaptic markers. Next, β‐GPA and American ginseng were tested in groups of explant cultures that were exposed to a lysosomal inhibitor (chloroquine) as a model for the age‐related compromise of lysosomal protein clearance capacity. Again, both β‐GPA‐ and American ginseng‐treated hippocampal explants displayed protection of pre‐ and postsynaptic proteins, similar to the protected levels produced through pharmacological enhancement of CatB using small‐molecule modulators with drug‐like properties (Viswanathan et al. 2012 – ACS Med Chem Lett 3:920). The significant results from the chloroquine model indicate that preventing age‐related lysosomal stress can slow or delay the associated synaptopathy by different strategies that may be additive. Together, the different experiments support the idea that exercise, exercise‐mimetics, and certain natural products help to limit age‐related synaptic deterioration that is known to be the best correlate of cognitive failure during aging and dementia. The common protective influences mediated by the exercise‐mimetic and the dietary supplement point to protein clearance as having a mechanistic role for healthy cognitive aging.