Culture Medium Research Articles

Trichomoniasis in Men: A Neglected Factor in Male Infertility?

This study is aimed to detect the frequency of trichomoniasis, a sexually transmitted infection caused by an anaerobic protozoan Trichomonas vaginalis, in men referred to the Fertility and Infertility Research Center Hamadan University of Medical Sciences. In this cross-sectional study, a group of 197 male volunteers who sought medical attention for issues related to infertility participated. The urine and semen samples were collected in sterile conditions. Both urine and semen sediment were promptly examined under a microscopy to detect the presence of motile trophozoites. Subsequently, 50µl of urine sediment were inoculated into the Dorset culture medium, whereas 50µl of semen sediment were inoculated into the Diamond culture (TYI-S-33) and incubated at 35.5oC. Finally, the processed urine samples were used for molecular analysis. The investigated subjects had an average age ranging from 36 to 40 years. There were 181 volunteers with fertility issues, outnumbering the 16 individuals with normal fertility. Spermogram analysis showed that 48% of participants had non-motile or progressive sperms, and 48% had abnormalities in sperm morphology. T. vaginalis was not detected through microscopic assessment, but PCR and sequence analysis revealed one case in a 33-year-old infertile individual, who had only 0.3% normal sperm with 19% motility. The isolated T.vaginalis was found to have the G genotype based on the sequencing analysis. Trichomoniasis in males was considered unimportant, with the belief that it would improve on its own. This study suggests that parasites can be considered as one of the male infertility factors, however, the impact is not fully understood. This investigation confirmed the effectiveness of molecular techniques in detecting trichomoniasis in males.

A xenogenic-free culture medium for cell micro-patterning systems as cell-instructive biomaterials for potential clinical applications

Cell micro-patterning controls cell fate and function and has potential for generating therapeutically usable mesenchymal stromal cell (MSC) populations with precise functions. However, to date, the micro-patterning of human cells in a translational context has been impossible because only ruminant media supplements, e.g. fetal bovine serum (FBS), are established for use with micro-patterns (MPs). Thus, there are currently no good manufacturing practice (GMP)-compliant media available for MPs. This study tested a xenogenic-free human plasma and platelet lysate (hP + PL) medium supplement to determine its compatibility with MPs. Unfiltered hP + PL medium resulted in significant protein deposition, creating a 'carpet-like' layer that rendered MPs ineffective. Filtration (3×/5×) eliminated this effect. Importantly, quantitative comparison using droplet digital PCR revealed that human MSCs in all media types exhibited similar profiles with strong myogenic Calponin 1/Transgelin 2 (TAGLN2) and weaker osteogenic alkaline phosphatase/Runt-related transcription factor 2 marker expression, and much weaker adipogenic (lipoprotein lipase/peroxisome proliferator-activated receptor gamma) and chondrogenic (collagen type II/aggrecan) expression, with profiles being dominated by myogenic markers. Within these similar profiles, an even stronger induction of the myogenic marker TAGLN2 by all hP + PL- compared to FBS-containing media. Overall, this suggested that FBS can be replaced with hP + PL without altering differentiation profiles. However, assessing individual MSC responses to various MP types with defined categories revealed that unfiltered hP + PL medium was unusable. Importantly, FBS- and 3× filtered hP + PL media were comparable in each differentiation category. Summarized, this study recommends 3× filtered hP + PL as a xenogenic-free and potentially GMP-compliant alternative to FBS as a culture medium supplement for micro-patterning cell populations in both basic and translational research that will ensure consistent and reliable MSC micro-patterning for therapeutic use.

High-Throughput Chemotherapeutic Drug Screening System for Gastric Cancer (Cure-GA).

Three dimensional (3D) cell cultures can be effectively used for drug discovery and development but there are still challenges in their general application to high-throughput screening. In this study, we developed a novel high-throughput chemotherapeutic 3D drug screening system for gastric cancer, named 'Cure-GA', to discover clinically applicable anticancer drugs and predict therapeutic responses. Primary cancer cells were isolated from 143 fresh surgical specimens by enzymatic treatment. Cell-Matrigel mixtures were automatically printed onto the micropillar surface then stabilized in an optimal culture medium for 3 days to form tumoroids. These tumoroids were exposed in the drug-containing media for 7 days. Cell viability was measured by fluorescence imaging and adenosine triphosphate assays. On average, 0.31±0.23g of fresh tumor tissue yielded 4.05×106 ± 4.38×106 viable cells per sample. Drug response results were successfully acquired from 103 gastric cancer tissues (success rate = 72%) within 13±2 days, averaging 6.4±2.7 results per sample. Pearson correlation analysis showed viable cell numbers significantly impacted drug data acquisition (p<0.00001). Tumoroids retained immunohistochemical characteristics, mutation signatures, and gene expression consistent with primary tumors. Drug reactivity data enabled prediction of synergistic drug correlations. Additionally, a multiparameter index-based prognosis model for patients undergoing gastrectomy followed by adjuvant XELOX was developed, showing significant differences in 1-year recurrence-free survival rates between drug responders and non-responders (p<0.0001). The Cure-GA platform enables rapid evaluation of chemotherapeutic responses using patient-derived tumoroids, providing clinicians with crucial insights for personalized treatment strategies and improving therapeutic outcomes.

Bisphenol A-Induced Cancer-Associated Adipocytes Promotes Breast Carcinogenesis Via CXCL12/AKT Signaling.

Bisphenol A (BPA), a commonly used plastic additive, is believed to cause obesity. As an environmental endocrine disruptor, BPA is closely associated with the onset and progression of BC. However, the molecular mechanisms underlying the promotion of breast cancer by BPA remain unclear. As obesity is a significant risk factor for breast cancer, this study aimed at exploring whether BPA facilitates the progression of breast cancer by inducing obesity. Using the National Health and Nutrition Examination Survey data, a positive correlation was observed between BPA exposure and the risk of sex-specific cancers among US adults with body mass index ≥30, suggesting that obesity may be influenced by urinary BPA. 3T3-L1 cells differentiated into mature adipocytes following treatment with 10-8 M BPA, and subsequent treatment with 4T1 culture medium acquired properties associated with cancer-associated adipocytes (CAAs). Network pharmacology suggested that CXCL12 may serve as a key target gene in breast cancer progression. Follow-up PCR analysis revealed high CXCL12 expression in BPA-induced CAAs. Overexpression of CXCL12 promoted epithelial-mesenchymal transition (EMT) and 4T1 cell migration by activating the AKT pathway. In vivo, BPA-induced CAAs accelerated tumor growth compared to a controls xenografted with only 4T1 cells. In tissues from the BPA-CAAs group, the expression of CXCL12, markers associated with CAAs, phosphorylated AKT, N-cadherin, and vimentin was markedly elevated, whereas the expression of E-cadherin was reduced. In conclusion, BPA may induce adipose cells to differentiate into CAA-like cells and subsequently advance breast cancer EMT through the CXCL12/AKT pathway.

Synergistic combination of orally available safe-in-man pleconaril, AG7404, and mindeudesivir inhibits enterovirus infections in human cell and organoid cultures

Enteroviruses can infect various human organs, causing diseases such as meningitis, the common cold, hand-foot-and-mouth disease, myocarditis, pancreatitis, hepatitis, poliomyelitis, sepsis, and type 1 diabetes. Currently, there are no approved treatments for enterovirus infections. In this study, we identified a synergistic combination of orally available, safe-in-man pleconaril, AG7404, and mindeudesivir, that at non-toxic concentrations effectively inhibited enterovirus replication in human cell and organoid cultures. Importantly, the cocktail did not alter glucose and insulin levels in the culture medium of pancreatic β-cells and preserved the contraction rhythm of infected heart organoids. These findings highlight a promising drug cocktail for further preclinical studies and clinical trials targeting a broad range of enterovirus-mediated diseases.

Hepatic Abnormal Secretion of Apolipoprotein C3 Promotes Inflammation in Aortic Dissection.

Apolipoprotein C3 (apo C3) is primarily secreted by the liver and is involved in promoting sterile inflammation and organ damage under pathological conditions. Previous studies have shown that apo C3 is abundant in the plasma exosomes of patients with aortic dissection (AD), but its specific role in AD remains unclear. In vivo, adeno-associated virus was used to knock down hepatic apo C3 expression in an AD mouse model to assess the impact of liver-derived apo C3 on the development of AD. Invitro, recombinant apo C3 protein was added to the culture medium of J774A.1 macrophages to evaluate its effect on macrophage polarization and to identify the underlying mechanisms. Additionally, the effect of apo C3 on the function of aortic endothelial and smooth muscle cells was explored. Apo C3 in the aortas of AD mice was found to originate from abnormal hepatic secretion, which enters the bloodstream and subsequently deposits in the aorta. Adeno-associated virus-mediated hepatic apo C3 knockdown significantly reduced AD incidence (P=0.0036), macrophage infiltration (P=0.0004), and collagen deposition in the aorta (P=0.0016). Similarly, inhibiting the apo C3 receptor Toll-like receptor 2 significantly lowered AD incidence (P=0.0352). Invitro, recombinant apo C3 protein promoted M1 polarization and matrix metalloproteinase secretion in macrophages by activating the Toll-like receptor 2/NLR family pyrin domain containing 3 pathway. Additionally, apo C3 increased adhesion molecule expression in endothelial cells and induced inflammation, chemotaxis, and apoptosis in vascular smooth muscle cells. Our findings highlight the role of abnormally secreted hepatic apo C3 in promoting aortic inflammation.

Nutrient control of splice site selection contributes to methionine addiction of cancer.

Many cancer cells depend on exogenous methionine for proliferation, whereas non-tumorigenic cells can divide in media supplemented with the metabolic precursor homocysteine. This phenomenon is known as methionine dependence of cancer or methionine addiction. The underlying mechanisms driving this cancer-specific metabolic addiction are poorly understood. Here we find that methionine dependence is associated with severe dysregulation of pre-mRNA splicing. We used triple-negative breast cancer cells and their methionine-independent derivatives R8 to compare RNA expression profiles in methionine and homocysteine growth media. The data set was also analyzed for alternative splicing. When tumorigenic cells were cultured in homocysteine medium, cancer cells failed to efficiently methylate the spliceosomal snRNP component SmD1, which resulted in reduced binding to the Survival-of-Motor-Neuron protein SMN leading to aberrant splicing. These effects were specific for cancer cells as neither Sm protein methylation nor splicing fidelity was affected when non-tumorigenic cells were cultured in homocysteine medium. Sm protein methylation is catalyzed by Protein Arginine Methyl Transferase 5 (Prmt5). Reducing methionine concentrations in the culture medium sensitized cancer cells to Prmt5 inhibition supporting a mechanistic link between methionine dependence of cancer and splicing. Our results link nutritional demands to splicing changes and thereby provide a link between the cancer-specific metabolic phenomenon, described as methionine addiction over 40 years ago, with a defined cellular pathway that contributes to cancer cell proliferation.

In vitro regeneration and optimization of physical and chemical mutagenesis protocol in tuberose (Agave amica (Medik.) Thiede & Govaerts) cv. ‘Arka Vaibhav’

Purpose Tuberose (Agave amica [Medik.]) is a vegetatively propagated commercial flower crop with limited genetic variability. Crossing barriers prevailing in tuberose necessitates modern breeding techniques like in vitro mutagenesis to generate variability. Hence, this study aimed to establish an efficient in vitro regeneration protocol for the rapid multiplication of tuberose and optimize the method for in vitro mutagenesis using the terminal stem scale as the explant. Results MS medium supplemented with 17.74 µM benzyl aminopurine) (BAP) and 0.57 µM indole-3-acetic acid (IAA) showed the maximum number of multiple shoots (5.0), with optimal shoot length (6.77 cm) and number of leaves (6.07). The shoots formed maximum rooting (99.44%) in MS medium supplemented with 4.90 µM indole-3-butyric acid, with an average of 26.89 roots per shoot. In vitro mutagenesis attempted physically via gamma irradiation led to an LD25, 50, 75 values of 13.21, 20.81, 32.79 Gy, respectively. The incorporation of ethyl methane sulfonate (EMS) into the culture media at a concentration of 0.08%, 0.13%, and 0.21% effectively resulted in LD25, 50, 75, respectively. Pretreating explants with 0.13% EMS for 15 min, 0.18% EMS for 30 min, 0.14% EMS for 45, and 0.11% EMS for 60 min were optimal for achieving 50% survival and plant regeneration using the regeneration protocol described above. Conclusion The regeneration protocol and optimized mutagen dose for in vitro mutagenesis developed in this study can be utilized for rapid multiplication of the cultivar and as a tool in genetic improvement programs aimed at inducing variability for commercially significant traits in tuberose.

Correlation of precisely fabricated geometric characteristics of DNA-origami nanostructures with their cellular entry in human lens epithelial cells

Human lens epithelial cells (hLECs) are critical for lens transparency, and their aberrant metabolic activity and gene expression can lead to cataract. Intracellular delivery to hLECs, especially to sub-cellular organelles (e.g., mitochondrion and nucleus), is a key step in engineering cells for cell- and gene- based therapies. Despite a broad variety of nano- and microparticles can enter cells, their spatial characteristics relevant to cellular uptake and localization remains elusive. To investigate cellular internalization of nanostructures in hLECs, herein, DNA nanotechnology was exploited to precisely fabricate four distinct, mass-controlled DNA-origami nanostructures (DONs) through computer-aided design. Ensembled DONs included the rods, ring, triangle, and octahedron with defined geometric parameters of accessible surface area, effective volume, compactness, aspect ratio, size and vertex number. Atomic force microscopy and agarose gel electrophoresis showed that four DONs self-assembled within 3.5h with up to 59% yield and exhibited structural intactness in cell culture medium for 4 h. Flow cytometry analysis of four Cy5-labelled DONs in hLECs HLE-B3 found time-dependent cellular uptake over 2 h, among which the octahedron and triangle had higher cellular accumulation than the rod and ring. More importantly, the vertex number among other geometric parameters was positively correlated with cellular entry. Confocal images further revealed that four DONs had preferential localization at mitochondria to nucleus at 2 h in HLE-B3 cells, and the degree of their biodistribution varied among DONs as evidenced by Manders’ correlation coefficient. This study demonstrates the DONs dependent cellular uptake and intracellular compartment localization in hLECs, heralding the future design of structure-modulating delivery of nanomedicine for ocular therapy.Graphical abstract

One carbon metabolism supplementation in maturation medium but not embryo culture medium improves the yield of blastocysts from bovine oocytes

Optimizing oocyte maturation and embryo culture media could enhance in vitro embryo production. The purpose of the present study was to investigate the role of supplementing one carbon metabolism (OCM) substrates and its cofactors (Cystine, Zinc, Betaine, B2, B3, B6, B12 and 5-methyltetrahydrofolate) in maturation and/or embryo culture media on the rate of blastocyst formation and pregnancy outcomes following the transfer of the resulting blastocysts in bovines. In the first experiment, 2537 bovine oocytes were recovered from slaughterhouse ovaries and then matured either in conventional maturation medium (IVM) or IVM supplemented with OCM substrates (Sup-IVM). After in vitro fertilization, the putative zygotes from each treatment (IVM or Sup-IVM) were cultured in the media either without (IVM/IVC or Sup-IVM/IVC) or with (IVM/Sup-IVC or Sup-IVM/Sup-IVC) OCM supplementation. The blastocyst rate, assessed on day 8, was significantly increased in Sup-IVM/IVC group (34.90 ± 2.52) as compared to IVM/IVC (17.06 ± 1.69; P = 0.0001) and Sup-IVM/Sup-IVC (20.29 ± 2.75; P = 0.004) and non-significantly as compared to IVM/Sup-IVC (24.86 ± 5.37). In the second experiment, non-matured bovine oocytes were collected by transvaginal ovum pick up after FSH stimulation, randomly allocated into IVM/IVC (n = 275) and Sup-IVM/IVC (n = 260) and the blastocysts achieved at day 7 were transferred in recipient cattle. The blastocyst rate was significantly higher in Sup-IVM/IVC group (38.85%) as compared to the IVM/IVC group (23.64%; P < 0.0001). After single embryo transfer, the supplemented blastocysts were at least as competent as non-supplemented ones with a non-significantly higher (20% vs. 14%) pregnancy rate and the advantage of several good quality blastocysts available for future use. In conclusion, optimizing the maturation medium with OCM substrates and its cofactors could enhance the formation of viable blastocysts with the potential to increase the cumulative birth rate in cattle.

Unlocking a Decade of Research on Embryo-Derived Extracellular Vesicles: Discoveries Made and Paths Ahead.

Over the past decade, research on embryo-derived extracellular vesicles (EVs) has unveiled their critical roles in embryonic development and intercellular communication. EVs secreted by embryos are nanoscale lipid bilayer vesicles that carry bioactive cargo, including proteins, lipids, RNAs, and DNAs, reflecting the physiological state of the source cells. These vesicles facilitate paracrine and autocrine signaling, influencing key processes such as cell differentiation, embryo viability, and endometrial receptivity. Studies reveal that EVs can traverse the zona pellucida, transferring molecular signals that enhance blastocyst formation and support embryo-maternal crosstalk. EVs have emerged as non-invasive biomarkers for embryo quality, with their cargo providing insights into genetic integrity and developmental competence. Advances in isolation and characterization techniques have identified specific microRNA (miRNAs) and transcription factors within EVs, offering potential for use in preimplantation genetic screening (PGS) and sex determination. Moreover, EV-mediated interactions with the maternal environment are critical for successful implantation, as they modulate gene expression and immune responses in endometrial and oviductal cells. Despite these advancements, challenges persist, including the standardization of EV isolation methods and the low yield of EVs DNA from spent culture media. Future research should aim to refine analytical techniques, explore EV-miRNA profiling, and investigate the mechanisms underlying EV-mediated signaling. By addressing these gaps, EVs could revolutionize embryo selection and reproductive technologies, offering new strategies to improve outcomes in assisted reproduction and animal breeding.

Anti-skin aging effects of Gosori liquor lees extract by regulating interactions between senescent fibroblasts and adipose-derived stem cells.

When cellular ageing is accelerated by various extrinsic/endogenous stimuli, regenerative function deteriorates, and enriched secretomes, such as the senescence-associated secretory phenotype (SASP), contribute to chronic inflammation and cause matrix degeneration. SASPs from senescent fibroblasts exacerbate cellular senescence via autocrine signalling and also accelerate skin ageing through the induction of neighbouring cell senescence via paracrine signalling. The interaction between dermis fibroblasts and their neighbours, adipose-derived stem cells (ADSCs) in the hypodermis, which lies deep in the dermis, is a potential target for skin ageing. In this study, we observed that an extract of the lees of Gosori liquor (GLE), a traditional Korean liquor made by fermenting millet and rice, suppressed the senescence of fibroblasts, including SASP production, in a replicative senescent model. We further examined whether the anti-ageing effects of GLE on fibroblasts affected the cellular senescence of their surrounding cells, ADSCs. The results showed that senescence factors in ADSCs were suppressed by culture medium from senescent fibroblasts (SF-CM) treated with GLE compared to the SF-CM-only treated group. Furthermore, the regenerative ability of ADSCs was promoted in the GLE-treated SF-CM group. ADSC migration was stimulated by upregulating the levels of α-smooth muscle actin, collagen type I alpha 2, and vascular endothelial growth factor expression through the PI3K/AKT pathway. Those results indicate that GLE can exert regenerative ability by regulating fibroblasts, and adipocyte interactions, improving cellular senescence. We conducted a clinical trial of subjects over 45 years of age to confirm the anti-ageing effects of GLE invivo and observed changes in ageing parameters, such as skin wrinkles and volume on the face (IRB No. DM-IRB-2023-809-01-T1). The results showed that GLE-containing cream was more effective in improving skin wrinkles, elasticity, density, thickness, and volume around sunken eyes after 4 weeks of use than placebo. In conclusion, GLE plays an important role in inhibiting the ageing transition to ADSCs by inhibiting the senescence of fibroblasts and can be a promising anti-ageing strategy.

Enhancement of the production of terpenoid and flavonoid secondary metabolites in the ground and aerial parts of licorice composite plant in a hydroponic system.

Hairy roots mediated by Agrobacterium rhizogenes can be obtained from the composite plants (plants with hairy roots and untransformed aerial parts) by ex vitro method. Composite plants can produce higher amounts of secondary metabolites by merging hydroponic systems. This provides a stable condition for composite plants, in which various metabolites are produced in different parts. In the present study, composite plants of Glycyrrhiza glabra were produced under ex vitro conditions and transferred into a hydroponic culture medium to produce and extract secondary metabolites. The results showed a 4.8- and 1.8-fold enhancement in the expressions of SQS1 and bAS genes in the roots of the composite plants compared to the control group, respectively. The levels of glycyrrhizin (1.7) and glabridin (3.5) were higher in the leaves of the composite plants compared to the controls. Moreover, higher amounts of glabridin (1.7) were observed in the roots of the composite plants compared to the control group. Investigation of the oxidative enzymes in the composite plants and control group revealed that the plants used more secondary metabolites through Agrobacterium inoculation. The plants needed more antioxidant enzymes to counter the release of oxygen-free radicals in control conditions, but composite plants used secondary metabolites as scavengers. Data revealed that composite plants managed to produce high amounts of various secondary metabolites in a hydroponic system.

Saponins enhance the stability and cost-efficiency of human embryonic stem cell culture

The cultivation and differentiation of human embryonic stem cells (hESCs) into organoids are crucial for advancing of new drug development and personalized cell therapies. Despite establishing of chemically defined hESC culture media over the past decade, these media's reliance on growth factors, which are costly and prone to degradation, poses a challenge for sustained and stable cell culture. Here, we introduce an hESC culture system(E6Bs) that facilitates the long-term, genetically stable expansion of hESCs, enabling cells to consistently sustain high levels of pluripotency markers, including NANOG, SOX2, TRA-1–60, and SSEA4, across extended periods. Moreover, organoids derived from hESCs using this medium were successfully established and expanded for at least one month, exhibiting differentiation into cortical organoids, GABAergic precursor organoids and heart-forming organoids. This innovative system offers a robust tool for preserving hESC homeostasis and modeling the nervous system in vitro.

Competing for Cultural Authority: Journalism Studies Must Account for the Right

ABSTRACT Recent technological and political developments have caused researchers to reevaluate journalists’ relevance vis a vis other forms of media production, with some calling for a “decentering” of journalism within broader media cultures. While there has been sizeable growth in research concerning the impacts of new digital technologies on journalism, we call for a renewed focus on partisan media. We contend that the future of journalism studies necessarily involves a reckoning with right-wing news cultures and note four areas where further research is especially necessary: including theorizing how conservative news influences and its epistemological assumptions, as well as right-wing media’s increasing salience across a diverse array of communities in the U.S. and across the globe. We intend this essay as a provocation to journalism studies scholars, with the goal of effecting a robust and reflexive subfield devoted to the study of right-wing news.

Optimization of Culture Medium Ingredients and Culture Conditions for Bacteriocin Production in Lactococcus lactis NCU036019.

Bacteriocin lactococcin036019 was identified and characterized from Lactococcus lactis NCU036019, which displayed significant antibacterial activity toward foodborne pathogenic bacteria Staphylococcus aureus under various conditions. However, the in situ low-level expression of lactococcin036019 severely limited its wide application in food industry. In this study, we optimized the medium ingredients and culture conditions of L. lactis NCU036019 for maximum production of lactococcin036019. The effects of different carbon sources, nitrogen sources, inorganic salts, growth factors, surfactants, and buffer salts on the production of bacteriocin were studied using antibacterial titer and diameter of inhibitory zone as evaluation indexes. Through single-factor experiments, Plackett-Burman (PB) experiment, steepest ascent experiment and response surface methodology, yeast extract, zinc sulfate, sodium acetate, mannitol, Tween-80, and dipotassium hydrogen phosphate were identified to display significant influence on the production of bacteriocin. By optimizing Man Rogosa and Sharpe (MRS) culture medium ingredients, the antibacterial activity of lactococcin036019 in the cell-free supernatant raised from 46.19 to 300.14Au/mL, namely, 6.5 times increased. Furthermore, the culture conditions, such as inoculation amount, culture time, and culture temperature, were optimized, and this further increased the antibacterial activity to 409Au/mL, namely, 8.8 times increased. This study investigated the effects of culture media and conditions on the production of lactococcin036019, and they were optimized for a maximum harvest of bacteriocin, and the significant increase of bacteriocin production in L. lactis NCU036019 facilitates the application of the antibacterial substance in future work.

High glucose induces pro-inflammatory polarization of macrophages by inhibiting immune-responsive gene 1 expression.

To investigate the effect of high glucose on macrophage polarization and the role of immune-responsive gene 1 (IRG1) in mediating its effect. RAW264.7 cells were transfected with IRG1-overexpressing plasmid or IRG1 siRNA via electroporation and cultured in either normal or high glucose for 72 h to observe the changes in cell viability and morphology using CCK-8 assay and phase contrast microscopy. The protein levels of IRG1, iNOS, Arg-1, IL-1β and IL-10 in the treated cells were detected with Western blotting, and the fluorescence intensities of iNOS and Arg-1 were detected using immunofluorescence assay. The protein levels of IL-1β and IL-10 in the culture medium were determined with ELISA. High glucose exposure significantly reduced IRG1 and Arg-1 expressions, increased iNOS and IL-1β expressions and IL-1β secretion, and decreased IL-10 level in RAW264.7 cells. Transfection with the IRG1-overexpressing plasmid provided the cells with obvious resistance to high glucose-induced changes in iNOS, Arg-1, IL-1β and IL-10, whereas IRG1 knockdown further enhanced the effects of high glucose exposure on Arg-1 expression and the expression and secretion of IL-10. High glucose promotes M1 polarization of the macrophages possibly through a mechanism to inhibit the expression of IRG1 protein, thus leading to chronic inflammatory response.

BMP6 participates in the molecular mechanisms involved in APAP hepatotoxicity.

Given the lack of accurate diagnostic methods of acetaminophen (APAP)-induced acute liver failure (ALF), the search for new biomarkers for its diagnosis is an urgent need. The aim of this study was to evaluate the role of bone morphogenetic protein 6 (BMP6) in APAP-induced ALF progression and its potential value as a biomarker of ALF. Hepatic and circulating BMP6 expression was assessed in APAP-treated mice and in serum samples from patients with APAP overdose. In addition, BMP6 expression and release was evaluated in hepatocytes after APAP exposure. BMP6 gene was silenced in Huh7 cells prior to APAP treatment and the culture medium (CM) was added to THP1 cells to evaluate the paracrine effects of hepatocyte BMP6 on APAP toxicity. Hepatic and serum BMP6 levels were increased in mice after APAP-induced ALF. In addition, a positive correlation was observed between circulating BMP6 and ALT activity in patients exposed to APAP overdose. Moreover, hepatocytes expressed and released BMP6 to the CM after APAP treatment. Indeed, the CM from APAP-treated Huh7 cells upregulated M1 and M2 markers in THP1 monocytes. The CM from BMP6-silenced Huh7, which was depleted of BMP6, reduced the expression of M2 markers in THP1 cells. In fact, expression of M2 markers was increased in THP1 cells exposed to BMP6. This study reveals that hepatic BMP6 expression is increased in APAP-induced acute liver injury, positioning it as a potential new biomarker of liver damage severity. Moreover, our data indicate that BMP6 might play a role in the hepatocyte-macrophage crosstalk during APAP-induced hepatotoxicity.

Label-free 3D cell imaging using hydrogels functionalized with switchable iridium complexes.

The use of fluorescent labels is the most common tool to visualize cells. However, the internalization of dye molecules often modifies the cell behavior. In this paper we demonstrate that it is possible to transiently label cells using a 3D scaffold, a hydrogel, covalently functionalized with luminescent cyclometalated iridium(III) complexes. The unique feature of our design is that the complexes are emissive only when they interact with the cell membrane while their emission is quenched in water. We exploited this feature to perform real-time and staining-free cell visualization and imaging. The hydrogels immerged in culture media are non-luminescent, however, when cells are added to it, they interact with the iridium complexes, covalently linked to the gel, and their lipophilic membrane "switches on" the luminescence enabling a clear and dynamic, real-time 3D visualization of cell proliferation. A complete photophysical and biological study of the materials is presented which demonstrates the potential of our methodology for 3D-realtime cell tracking.

Contribution to the Identification and Study of the Pathogenicity of Bacteria Responsible for Cassava Root Rot in Côte d'Ivoire Production Zones

Background and Objective: Cassava is an important crop for the Ivorian population. However, yields are relatively low. This low productivity is due to cassava root rots, which are sometimes caused by bacteria. The general objective was to identify the main bacterial diseases responsible for cassava tuberous root rot in production areas in Côte d'Ivoire. Specifically, to identify pathogenic bacteria and then to assess the pathogenicity of the bacterial strains isolated. Material and Methods: The plant material consisted of tuberous cassava roots affected by rot. Decay-causing bacteria were isolated on YPGA (Yeast extract Peptone Glucose Agar) culture medium. Strain identification was carried out using the API 20 E and 20NE gallery. Tuberous cassava roots were perforated with a punch and inoculated with bacterial inoculum calibrated to an optical density of 0.2 at a wavelength of 600 nm, corresponding to 108. Results: 174 strains were isolated. The highest proportions of strains were obtained in the localities of Ferké and Man, with 19 and 26% respectively. The bacterial species identified were Roualtella planticola, Serratia liquefaciens, Serratia marcescens, Burkholderia cepacia, Chromobacterium violaceum, Erwinia spp, Enterobacter cloacae, Klebseilla pneumoniae spp ozaenae and Raoutellia ornithinolytica. The Erwinia species was the most frequent in five localities, with proportions of 100 and 71.4% respectively in Agboville and Yamoussoukro. Results showed that 91.5% of bacterial strains tested caused rot (soft or dry). On the other hand, 52.4% of bacterial strains induced dry rot and around 39% induced soft rot. Conclusion: Various bacterial species were identified in this study. In addition, these species vary in their germinative capacity.