Differentiation Medium Research Articles

Bactericidal versus bacteriostatic antibacterials: clinical significance, differences and synergistic potential in clinical practice

Abstract Antibacterial activity can be classified as either bactericidal or bacteriostatic, using methods such as the MBC/MIC ratio and time–kill curves. However, such categorization has proven challenging in clinical practice, as these definitions only apply under specific laboratory conditions, which may differ from clinical settings. Several factors, such as the specific bacteria or infectious medium, can affect the action of antibiotics, with many antibacterials exerting both activities. These definitions have also led to the belief that bactericidal antibacterials are superior to bacteriostatic, especially in more severe cases, such as endocarditis, neutropenia and bacteraemia. Additionally, current dogma dictates against the combination of bactericidal and bacteriostatic antibacterials in clinical practice, due to potential antagonism. This review aimed to assess the differences in antibacterial activity of bactericidal and bacteriostatic antibacterials based on in vitro and in vivo studies and examine their antagonistic or synergistic effects. Our findings show that specific bacteriostatic agents, such as linezolid and tigecycline, are clinically non-inferior to bactericidals in multiple infections, including pneumonia, intra-abdominal infections, and skin and soft tissue infections. Studies also support using several bacteriostatic agents as salvage therapies in severe infections, such as neutropenic fever and endocarditis. Additionally, not all combinations of bacteriostatic and bactericidal agents appear to be antagonistic, with many combinations, such as linezolid and rifampicin, already being used. The findings should be interpreted with caution, as most evidence is from observational studies and there is a need for randomized controlled trials to assess their effectiveness and combinations, especially within the context of rising antimicrobial resistance.

Using marketing strategies to promote maternal health: an international students’ collaboration

Abstract Purpose About 1 in 8 birth parents experience symptoms of perinatal mood and anxiety disorders (PMADs). The purpose of this collaborative project was to create a social media deliverable to lower the risk of postpartum depression in pregnant and postpartum parents in Georgia, USA. Methods Using the SM 10-step model, Master of Public Health Students from Georgia Southern University (GSU), Georgia in collaboration with Creative Business students from Breda University of Applied Science (BUas), the Netherlands, designed an evidence-based SM product. They were mentored by public health and media faculty from both institutions. Roles and responsibilities according to the skill sets were divided, but briefly: students 1) identified a target audience; 2) conducted tertiary data collection; 3) set goals and objectives; 4) developed 4 P’s (product, price, place, and promotion) 5) developed marketing strategies.; 6) and used the strategy to create a specific social media deliverable. Results Utilizing the 10-step model social marketing strategy used state-of-the-art marketing and digital media techniques and evidence-based strategies to create a digital electronic booklet that focused on reducing the stigma associated with help seeking behaviors with the ultimate goal to reduce the incidence and prevalence of PMADs. The digital interactive booklet will be pilot tested in selected OBGYN offices and health departments in Southeast Georgia in July 2024. Conclusions This collaborative project demonstrates effective collaboration between disciplines from different world regions utilizing strengths from both teams: public health and digital media skills to create an SM product that will be utilized to promote social engagement among postpartum parents. Key messages • Students utilized social marketing strategies to create an electronic booklet to address postpartum depression using state-of-the-art digital media techniques and evidence-based strategies. • This collaborative project demonstrates effective collaboration between disciplines from different world regions and utilizes strengths from both teams.

THE PREVALENCE OF (OMPA, CSUE) GENES AMONG BIOFILM PRODUCER ACINETOBACTER BAUMANNII ISOLATES

Acinetobacter baumannii is a significant public health problem because it is capable of forming biofilms that may be responsible for the survival of this pathogen in the hospital environment. The aim of this study is to determine the role of (OmpA, CsuE) genes in biofilm production among A. baumannii isolates. A total of 100 clinical isolates of bacteria were collected from a different sources. All isolates were identified by biochemical test, specific selective media (CHROMagar) and polymerase chain reaction by detection presence of (blaOXA51) gene. only 60 isolates were diagnosed as A. baumannii, and to determine the biofilm production capacity two different methods were used Congo red agar method(CRA) and Microtiter plate method (MTP). MTP method result showed 85% of isolates were able to form biofilms, on the other hand, 33.33% of isolates had a biofilm production ability by showing bright black colonies on CRA. Conventional PCR was used to detect the presence of (OmpA, CsuE) genes and the result showed the prevalence of both genes in all isolates were 97%(58/60). It also shows a strong correlation between the presence of these genes and biofilm formation.

Innovative 3D-Printed Titanium Specimens Favor a Modulation of Inflammation in Dental Pulp Stem Cells During Liposome-Triggered Mineralization.

developing customized titanium specimens, with innovative surfaces, is a suitable strategy to overcome implant failure. Additionally, a faster and efficient osteogenic commitment assists tissue regeneration. To investigate the interplay between inflammation and differentiation upon implantation, Dental Pulp Stem Cells (DPSCs) were cultured on 3D-printed titanium owning an internal open cell form, administering osteogenic factors by a liposomal formulation (LipoMix) compared to traditional delivery of differentiation medium (DM). osteogenic differentiation was evaluated by western blot, by measuring β1 integrin expression, and by and real-time RT-PCR, by measuring SP7 and Collagen I gene expression; while.angiogenesis was characterized by measuring VEGF secretion levels. Matrix mineralization was assessed by means of Alizarin Red Staining, cell adhesion and inflammation responses through western blot, enzymatic and ELISA assays evaluating Nrf2 expression, catalase activity and Prostaglandin E2 secretion, respectively. LipoMix enhances cell proliferation and adhesion, as revealed by increased integrin β1 expression. Mineralized matrix deposition, SP7 gene expression, Collagen I release and Alkaline Phosphatase activity appear increased in LipoMix condition. Additionally, the redox-sensitive transcription factor Nrf2 is overexpressed at the earliest experimental times, triggering the catalase activity. data reported confirm that internal topography and post-production treatments on titanium surfaces dynamically and positively condition the DPSC progress towards the osteogenic phenotype, moreover, the combination with LipoMix fastens the positive modulation of inflammation under osteogenic conditions. Therefore, the development of customized surfaces along with the administration of differentiating factors enclosed in a liposomal delivery system, could represent a promising and innovative tool in regenerative dentistry.

High-Density Fermentation of Lactobacillus plantarum P6: Enhancing Cell Viability via Sodium Alginate Enrichment

Lactobacillus plantarum exhibits a wide range of beneficial physiological functions, including maintaining intestinal microbiota balance, reducing serum cholesterol, and promoting digestive health. According to the specific nutrient requirements of Lactobacillus plantarum P6, we investigated the effects of various carbon sources, nitrogen sources, trace elements, growth-promoting substances, as well as the initial pH and inoculum size on the growth of Lactobacillus plantarum P6 under fermentation conditions. The optimal growth conditions for Lactobacillus plantarum P6 were identified to facilitate high-density fermentation in small-scale fermenter production, achieving a cell concentration of 1.03 × 1011 CFU/mL. This resulted in a 2.5-fold increase in bacterial wet weight, and fermentation time was reduced to 12 h when utilizing a specific medium enriched with 0.2% sodium alginate. It is hypothesized that sodium alginate forms a protective film around the bacterial cells, promoting cell aggregation and enhancing self-coalescence, potentially triggering a bacterial community effect. These results provide a basis for the industrial-scale high-density cultivation of Lactobacillus plantarum, offering potential for enhanced biotechnological applications.

Sodium danshensu modulates skeletal muscle fiber type formation and metabolism by inhibiting pyruvate kinase M1

Sodium Danshensu (SDSS) is extracted from Salvia miltiorrhiza and has many pharmacological effects. However, little is known about its effects on muscle fiber formation and metabolism. Here, we aimed to investigated the role and molecular mechanisms of SDSS in modulating the formation of skeletal muscle fiber. C2C12 cells were incubated in differentiation medium with or without SDSS for 4 days. C57BL/6 mice were orally administered SDSS by gavage once a day for 8 weeks. Grip strength, treadmill, muscle weight, western blotting, qPCR, immunofluorescence staining and H&E staining were performed. SDSS target proteins were searched through drug affinity responsive target stability (DARTS) and mass spectrometry analysis. Furthermore, molecular docking was carried out for Pyruvate kinase M1 (PKM1). The effect of PKM1 on myosin heavy chain (MyHCs) gene expression was verified by knockdown of PKM1 experiment. SDSS induced oxidative muscle fiber-related gene expression, and inhibited glycolytic fiber-related gene expression in C2C12 cells. Muscle mass, the percentage of slow oxidative fibers, succinic dehydrogenase activity, muscle endurance, glucose tolerance, and the expression of the MyHC1 and MyHC2a genes increased while MyHC2b expression, lactate dehydrogenase activity, and the percentage of glycolytic muscle fibers decreased in SDSS-treated mice. Mechanistically, SDSS bound to the pyruvate kinase PKM1 and significantly repressed its activity. PKM1 inhibited MyHC1 and MyHC2a expression but promoted MyHC2b expression. SDSS also significantly attenuated the effects of PKM1 on muscle fiber-related gene expression in C2C12 cells. Our findings indicate that SDSS promotes muscle fiber transformation from the glycolytic type to the oxidative type by inhibiting PKM1 activity, which provide a new idea for treating muscle atrophy, muscle metabolism diseases and improving animal meat production.

Israel in the Italian Media Before 7 October

Images broadcast on television, radio analyses, and long articles printed in newspapers and, today, the content of digital platforms have become an overwhelming part of our daily lives. The history of the State of Israel, which has been debated since its foundation, is very complex and not exempt from the reverberations that the policies of the opposing blocs have had over time on the conflicts that followed. Given the attention of the European public to international events concerning Israel and the role of the media in conveying information about them, the aim of this research was to investigate how Israel is talked about in the current media environment in Italy, particularly the digital one, within the Facebook platform between the years 2019 and 2021. This is the period before the outbreak of the war on 7 October 2023; this is to try to understand whether Israel has always been narrated according to specific media frames in Italy, such as those we see emerging with the current conflict against Hamas, or is it possible to highlight different aspects in previous years within the same context of crisis, but a global health crisis: before and after COVID-19.

The Austrian Political Advertisement Scandal: Patterns of “Journalism for Sale”

Mounting concern surrounds the influence of political actors on journalism, especially as media outlets face increasing financial pressures. These circumstances can give rise to instances of media capture, a mutually corrupting relationship between political actors and media organizations. However, empirical evidence substantiating such mechanisms and their consequences remains limited, particularly in the context of Western democracies. This chapter investigates a recent case in which a former Austrian chancellor allegedly colluded with a tabloid newspaper to receive better news coverage in exchange for increased ad placements by government institutions. We employ automated content analysis to investigate political news articles from seventeen prominent Austrian news outlets spanning 2012–2021 ( n = 222,659). Adopting a difference-in-differences approach, we find a substantial increase in media visibility of the former Austrian chancellor within the news outlet that is alleged to have received bribes, as well as a decrease in favorability for challenger candidates. Although this study does not aim to prove or disprove the involvement of specific political actors or media organizations in unethical or illegal activities, it introduces an innovative method for detecting unusual patterns in media reporting. Findings are discussed in the context of current threats to media independence and underscore the crucial need to protect journalistic integrity and ensure unbiased information for the public.

Diversity and Antifungal Susceptibilities of Yeasts from Mangroves in Hong Kong, China-A One Health Aspect.

While mangrove ecosystems are rich in biodiversity, they are increasingly impacted by climate change and urban pollutants. The current study provides first insights into the emergence of potentially pathogenic yeasts in Hong Kong's mangroves. Sediment and water samples were collected from ten urban and rural mangroves sites. Initial CHROMagarTM Candida Plus screening, representing the first application of this differential medium for water and soil samples collected from a non-clinical environment, enabled the rapid, preliminary phenotypic identification of yeast isolates from mangroves. Subsequent molecular profiling (ITS and/or 28S nrDNA sequencing) and antifungal drug susceptibility tests were conducted to further elucidate yeast diversity and drug resistance. A diversity of yeasts, including 45 isolates of 18 distinct species across 13 genera/clades, was isolated from sediments and waters from Hong Kong mangroves. Molecular profiling revealed a dominance of the Candida/Lodderomyces clade (44.4%), a group of notorious opportunistic pathogens. The findings also reveal a rich biodiversity of non-Candida/Lodderomyces yeasts in mangroves, including the first reported presence of Apiotrichum domesticum and Crinitomyces flavificans. A potentially novel Yamadazyma species was also discovered. Remarkably, 14.3% of the ubiquitous Candida parapsilosis isolates displayed resistance to multiple antifungal drugs, suggesting that mangroves may be reservoirs of multi-drug resistance. Wildlife, especially migratory birds, may disseminate these hidden threats. With significant knowledge gaps regarding the environmental origins, drug resistance, and public health impacts of pathogenic yeasts, urgent surveillance is needed from a One Health perspective. This study provides an early warning that unrestrained urbanization can unleash resistant pathogens from coastal ecosystems globally. It underscores the necessity for enhanced surveillance studies and interdisciplinary collaboration between clinicians, ornithologists, and environmental microbiologists to effectively monitor and manage this environmental health risk, ensuring the maintenance of 'One Health'.

Placental microphysiological systems: new advances on promising platforms that mimic the microenvironment of the human placenta.

One of the most complex human physiological processes to study is pregnancy. Standard animal models, as well as two-dimensional models, lack the complexity and biological relevance required to accurately study such a physiological process. Recent studies have focused on the development of three-dimensional models based on microfluidic systems, designated as placental microphysiological systems (PMPSs). PMPS devices provide a model of the placental barrier through culturing relevant cell types in specific arrangements and media to mimic the in vivo environment of the maternal-fetal circulation. Here, recent developments of PMPS models for embryo uterine implantation, preeclampsia evaluation, and toxicological screening are presented. Studies that use bioprinting techniques are also discussed. Lastly, recent developments in endometrium microphysiological systems are reviewed. All these presented models showed their superiority compared to standard models in recapitulating the biological environment seen in vivo. However, several limitations regarding the types of cells and materials used for these systems were also widely reported. Despite the need for further improvements, PMPS models contribute to a better understanding of the biological mechanisms surrounding pregnancy and the respective pathologies.

“journalists are just employees”: Examining Journalistic Autonomy from an Audience Perspective

ABSTRACT Autonomy is a fundamental requirement for the professional practice of journalism. However, while its role in ensuring well-informed audiences is widely acknowledged, the way in which audiences make sense of journalists’ autonomy remains unexplored. Similarly, the understanding of how audience perceptions of journalists’ autonomy, their attributed professionalism and authority, and the resulting impact on journalism practice remains incomplete. To address these gaps, this study conducts a qualitative analysis involving semi-structured interviews with 64 news users in Chile. Results show that journalists’ autonomy, from audiences’ view, is limited by labor forces and organizational influences. Journalists’ working conditions are perceived to be connected to high levels of editorial control, yielding a deficiency of autonomy that relegates journalists to a subservient role, compromising their professionalism, undermining their authority, and eroding the quality of the information they provide. Also, findings show that autonomy assumes a prominent role as a distinct form of authority when journalists diverge from conventional news norms, establishing a closer connection with their audience. Furthermore, independent journalists frequently garner recognition for their autonomy and credibility, attributed to their detachment from specific media outlets. The article delves into the implications of these findings for journalism studies and practice.

Fluid effect on elastic properties of carbonate and implication for CO2 geological sequestration monitoring

Laboratory measurement of the elastic properties of carbonates saturated with different fluids is crucial for characterizing reservoirs and monitoring CO2 sequestration. However, it is challenging to distinguish the effects of different fluids. We measured eight carbonate samples under varying pressure and fluids saturation conditions, and conducted quantitative analysis. The measurements showed that water saturation can increase P-wave velocity (Vp), while oil and CO2 saturation may lead to an increase, decrease or no change in Vp. Meanwhile, all fluids saturation reduces the S-wave velocity (Vs). The velocities of dolomite samples are higher and less pressure-sensitive than that of limestone samples. Four attributes are adopted to distinguish the fluids effect, including elastic moduli, impedance, Vp/Vs ratio, and a newly designed fluid discrimination factor F, which is proposed by multiplying P-wave modulus and square value of density change. Among the attributes, the F factor performed the best as it achieved evident and consistent results for all the samples. Rock physics models were used to simulate the elastic behaviors with the differential effective medium model predicting Vs the best. The Gassmann’s equation correctly indicated the positive or negative changes of rock velocity after fluid saturation even though its prediction did not exactly match the data. This study highlights the impact of various pore fluids on the carbonate elastic properties through laboratory measurement and theoretical modeling, and introduces a novel fluid identification factor, providing insights for reservoir seismic characterization and CO2 sequestration monitoring.

Genome sequencing analysis of the pncA, rpsA and panD genes responsible for pyrazinamide resistance of Mycobacterium tuberculosis from Indonesian isolates.

Developing the most suitable treatment against tuberculosis based on resistance profiles is imperative to effectively cure tuberculosis patients. Whole-genome sequencing is a molecular method that allows for the rapid and cost-effective detection of mutations in multiple genes associated with anti-tuberculosis drug resistance. This sequencing approach addresses the limitations of culture-based methods, which may not apply to certain anti-TB drugs, such as pyrazinamide, because of their specific culture medium requirements, potentially leading to biased resistance culture results. Thirty-four M. tuberculosis isolates were subcultured on a Lowenstein-Jensen medium. The genome of these bacteria was subsequently isolated using cetyltrimethylammonium bromide. Genome sequencing was performed with Novaseq Illumina 6000 (Illumina), and the data were analysed using the GenTB and Mykrobe applications. We also conducted a de novo analysis to compare the two methods and performed mutation analysis of other genes encoding pyrazinamide resistance, namely rpsA and panD. The results revealed mutations in the pncA gene, which were identified based on the databases accessed through GenTB and Mykrobe. Two discrepancies between the drug susceptibility testing and sequencing results may suggest potential instability in the drug susceptibility testing culture, specifically concerning PZA. Meanwhile, the results of the de novo analysis showed the same result of pncA mutation to the GenTB or Mykrobe; meanwhile, there were silent mutations in rpsA in several isolates and a point mutation; no mutations were found in the panD gene. However, the mutations in the genes encoding pyrazinamide require further and in-depth study to understand their relationship to the phenotypic profile. Compared to the conventional culture method, the whole-genome sequencing method has advantages in determining anti-tuberculosis resistance profiles, especially in reduced time and bias.

Peer-to-Peer Human Milk Sharing Through Social Media Groups: The Experiences of Women Donors and Recipients in Hong Kong.

To explore the lived experience of donors and recipients involved in peer-to-peer human milk donation. An exploratory descriptive qualitative study. Semi-structured interviews were conducted between November 2022 and April 2023. A total of 50 women consisting of 34 donors and 16 recipients were recruited using purposive sampling from specific social media groups in Hong Kong that focus on breast milk donation and sharing. Data were analysed based on thematic analysis. Five key themes were identified in this study: Motivation; Milk sharing as a social act; Not a Norm; Recommendation to formal milk banks; and Formal donation as the preferred route. The findings indicate informal milk sharing is seen as an inferior alternative to formal milk donation systems. The establishment of formal milk banks is viewed as a way to address the shortcomings of informal sharing. The findings have implications for the healthcare field, particularly in improving the experiences of those involved in informal milk sharing. The insights gained from the experiences of informal milk sharers can directly inform and enhance the services provided by formal milk banks. Given the growing prevalence of online informal milk sharing, healthcare professionals should enhance their understanding of this practice. However, the practice has remained understudied, particularly in Asian region. This study brings together the experiences of both milk donors and recipients, providing a comprehensive view on the phenomenon. The Standards for Reporting Qualitative Research (SRQR). No patient or public contribution.

Analisis Media Statement Kasus Lion Air Penerbangan Jt-610 Boeing 737 Max 8 Dengan Pendekatan Storytelling Michael L. Kent

The focus of this study is on how Lion Air managed crisis communication following the Boeing 737 MAX 8 crash of Flight JT-610. The research delves into the use of storytelling in media statements as a strategy for crisis communication during the aftermath of the crash. It outlines Lion Air's goals in sharing information and engaging with the public and media in the midst of this crisis. The study utilizes a qualitative text analysis to scrutinize three specific media statements released during the crisis. By applying Michael L. Kent's storytelling elements, the analysis of the media statements concentrates on factors like emplotment, narrative theory, identification, form, and masterplots. The selected statements cover the immediate aftermath of the crash on October 29, 2018; a subsequent statement on March 13, 2019, addressing broader safety concerns surrounding the Boeing 737 MAX 8; and a final statement on October 26, 2019, summarizing findings from the National Transportation Safety Committee's investigation. The research findings suggest that employing storytelling through straightforward text in media statements, incorporating credible character depiction, structured narrative, empathy, and compelling storytelling, can notably influence an organization's reputation recovery and improvement. Well-crafted and timely media releases are essential as primary information sources for the media, especially in crisis scenarios where prompt and precise information dissemination is critical. This method ensures that information, though disseminated quickly, maintains relevant and impactful storytelling aspects. The study concludes that storytelling, characterized by humanistic, empathetic, and coherent narratives across media statements, can be effectively utilized by Lion Air to uphold consistent and narrative-focused communication with the public through the media.

9194 Generation of a Human-Induced Pluripotent Stem Cell Line From a Patient With Hypopituitarism and a Novel Nonsense Variant in FOXA2

Abstract Disclosure: M.A. Camilletti: None. G.T. Chirino Felker: None. G. Amin: None. S. Castañeda: None. F. Zabalegui: None. C. Romano Florit: None. A. Waisman: None. M. Ciaccio: None. M.I. Di Palma: None. E. Vaiani: None. A. Belgorosky: None. S.G. Miriuka: None. L.N. Moro: None. M.I. Perez-Millan: None. Congenital Hypopituitarism (CH) is a genetically complex disease characterized by one or more pituitary hormone deficiencies. Forkhead Box A2 (FOXA2, 600288) is a pioneer transcription factor associated with development of multiple tissues in humans and mice. Heterozygous loss of function mutations in FOXA2 were recently shown to cause CH with hyperinsulinemic hypoglycemia, but little is known about the molecular mechanism of FOXA2 action during pituitary development. We identified a novel, heterozygous nonsense variant in FOXA2 (c.686C<A; p.S229X) in a patient diagnosed with GH deficiency at 1.7 years of age, with anterior lobe hypoplasia and absent pituitary stalk, asymptomatic hypoglycemia, and craniofacial malformations. To discover the mechanism of FOXA2 regulation of pituitary development we generated an induced pluripotent stem cell line (FOXA2mut-iPSC) from the patient’s polymorphonuclear blood cells (PBMCs) using an EF1a-hSTEMCCA-loxP vector containing the pluripotency genes OCT-4, SOX-2, c-MYC and KLF4. FOXA2mut-iPSC clone R1 had a normal karyotype and embryonic stem cell-like morphology. These cells express endogenous pluripotency-associated markers NANOG, SOX2, and OCT4, have alkaline phosphatase activity, and can differentiate into all three developmental layers, indicating that these cells are pluripotent. Short tandem repeat (STR) analysis confirmed that FOXA2mut-iPSC R1 clone profiles matched those of the donor PBMCs. Next, we compared pituitary differentiation of FOXA2mut-iPSC vs. control-iPSCs in vitro. Cells were cultured in a differentiation medium containing BMP4, the smoothened agonist SAG, and fibroblast growth factor (FGF), as described in Zimmer et al., 2016 (1). Gene expression analysis (by qRT-PCR) of samples collected on days 0, 4, 7, and 15 of the differentiation protocol showed an increased expression of genes associated with anterior pituitary development including OTX2, PITX1/2, and SIX1 in both cell lines (n=2). These preliminary data suggest that pituitary cell fate can be induced in the heterozygous FOXA2mut-iPSC clone. Ongoing studies will assess pituitary hormone-producing cell differentiation after longer periods of FOXA2mut-iPSC cell culture. In sum, patient iPSC differentiation is a promising tool for assessment of normal and variant FOXA2 function in human pituitary development, and for enlightening the mechanisms of FOXA2 action in human pituitary development. (1) Zimmer et al., Stem Cell Reports. 2016 Jun 14;6(6):858-872. Presentation: 6/1/2024

7151 Generation of a Human-Induced Pluripotent Stem Cell Line From a Patient With Hypopituitarism and a Novel Nonsense Variant in FOXA2

Abstract Disclosure: M.A. Camilletti: None. G.T. Chirino Felker: None. G. Amin: None. S. Castañeda: None. F. Zabalegui: None. C. Romano Florit: None. A. Waisman: None. M. Ciaccio: None. M.I. Di Palma: None. E. Vaiani: None. A. Belgorosky: None. S.G. Miriuka: None. L.N. Moro: None. M.I. Perez-Millan: None. Congenital Hypopituitarism (CH) is a genetically complex disease characterized by one or more pituitary hormone deficiencies. Forkhead Box A2 (FOXA2, 600288) is a pioneer transcription factor associated with development of multiple tissues in humans and mice. Heterozygous loss of function mutations in FOXA2 were recently shown to cause CH with hyperinsulinemic hypoglycemia, but little is known about the molecular mechanism of FOXA2 action during pituitary development. We identified a novel, heterozygous nonsense variant in FOXA2 (c.686C<A; p.S229X) in a patient diagnosed with GH deficiency at 1.7 years of age, with anterior lobe hypoplasia and absent pituitary stalk, asymptomatic hypoglycemia, and craniofacial malformations. To discover the mechanism of FOXA2 regulation of pituitary development we generated an induced pluripotent stem cell line (FOXA2mut-iPSC) from the patient’s polymorphonuclear blood cells (PBMCs) using an EF1a-hSTEMCCA-loxP vector containing the pluripotency genes OCT-4, SOX-2, c-MYC and KLF4. FOXA2mut-iPSC clone R1 had a normal karyotype and embryonic stem cell-like morphology. These cells express endogenous pluripotency-associated markers NANOG, SOX2, and OCT4, have alkaline phosphatase activity, and can differentiate into all three developmental layers, indicating that these cells are pluripotent. Short tandem repeat (STR) analysis confirmed that FOXA2mut-iPSC R1 clone profiles matched those of the donor PBMCs. Next, we compared pituitary differentiation of FOXA2mut-iPSC vs. control-iPSCs in vitro. Cells were cultured in a differentiation medium containing BMP4, the smoothened agonist SAG, and fibroblast growth factor (FGF), as described in Zimmer et al., 2016 (1). Gene expression analysis (by qRT-PCR) of samples collected on days 0, 4, 7, and 15 of the differentiation protocol showed an increased expression of genes associated with anterior pituitary development including OTX2, PITX1/2, and SIX1 in both cell lines (n=2). These preliminary data suggest that pituitary cell fate can be induced in the heterozygous FOXA2mut-iPSC clone. Ongoing studies will assess pituitary hormone-producing cell differentiation after longer periods of FOXA2mut-iPSC cell culture. In sum, patient iPSC differentiation is a promising tool for assessment of normal and variant FOXA2 function in human pituitary development, and for enlightening the mechanisms of FOXA2 action in human pituitary development. (1) Zimmer et al., Stem Cell Reports. 2016 Jun 14;6(6):858-872. Presentation: 6/1/2024

Zwitterionic MOF-embedded alginate beads with polydopamine surface functionalization for efficient doxycycline removal: Optimization and mechanistic study

The adsorptive removal of amphoteric antibiotics like doxycycline (DOX) is a difficult task because of the electrostatic repulsion between these amphoteric molecules and adsorbents. For this purpose, a zwitter adsorbent was fabricated by incorporating zwitter ZIF-67/MIL-88A binary MOF into the matrix of alginate (Alg); in addition, the surface of the beads was modified by polydopamine (PDA). The batch experiments implied the super-high adsorption efficacy of ZIF-67/MIL-88A@Alg@PDA toward DOX attained 384.61 ± 5.08 mg/g at a neutral pH medium, 25 °C, and using 0.02 g. The isotherm analysis implied the physisorption of DOX onto ZIF-67/MIL-88A@Alg@PDA, while the kinetic analysis denoted the chemisorption of DOX. The results of XPS, Zeta potential, and Lab experiments identified the types of physical and chemical interactions between ZIF-67/MIL-88A@Alg@PDA and DOX. The durability of the ZIF-67/MIL-88A@Alg@PDA beads was inspected by the recycling test, clarifying that the DOX adsorption aptitude declined by 12.22 mg/g. In addition, the measured leaching concentrations of cobalt and iron from the leaching test were 0.008 and 0.098 mg/L. The ionic strength of ZIF-67/MIL-88A@Alg@PDA, implying an enhancement in the DOX removal (%) from 83.51 to 93.50 % by raising the NaCl concentration from 0.2 to 1.0 mol/L. Therefore, our study could provide a simple procedure to overcome the electrostatic repulsion that retard the adsorption process of the amphoteric drugs onto charged adsorbents with positive or negative charges. Additionally, this procedure could also generate an electrostatic interaction between the zwitter adsorbents and the amphoteric drugs at specific pH media when they are in a zwitterionic nature.

Numerical investigation on the core thermal hydraulic behavior of pool-type sodium-cooled fast reactor (SFR)

A thorough understanding of the reactor core thermal hydraulic behavior is essential for the design and safety analysis of Sodium-cooled Fast Reactors (SFR). Due to the application of hexagonal subassembly, the core thermal hydraulic behavior is significantly affected by the flow field within the subassemblies, the inter-wrapper region and the hot pool. Analysis of the core thermal hydraulic behavior requires a model coupling the three regions mentioned above, which has been identified as one of the thermal hydraulic challenges in SFR. In the present study, a 3D model that covers the three regions was developed for the core of the China Experimental Fast Reactor (CEFR) with the Computational Fluid Dynamic (CFD) code, Fluent. The inter-wrapper region and the hot pool were modeled in detail, while the subassemblies were modeled with a special porous medium model. The core thermal hydraulics behavior under steady state was studied, more specifically, information for the flow field distribution at the core outlet, the inter-wrapper flow and the duct wall temperature distribution was obtained. Under steady state, liquid sodium in the inter-wrapper region is supplied by the inner region of the hot pool. And it enters the inter-wrapper region from the core outer region and returns back to the hot pool inner region from the core central region. The inter-wrapper flow is cooled by non-fuel subassemblies and heated up by fuel subassemblies. For non-fuel subassembly, the ratio of the total heat transfer rate between the inter-wrapper flow and the subassemblies to the heat generated within subassemblies could reaches 96%; for fuel subassemblies, the maximum ratio of the total heat transfer rate between the inter-wrapper flow and the subassemblies to the heat generated within subassemblies is 2.45%. Significant temperature gradients have been observed on the duct wall, with maximum values of 156.69 K/m in the vertical direction and 2,196.00 K/m in the circumferential direction. The largest temperature gradient appears on the duct of subassemblies adjacent to the transition region of fuel subassemblies and non-fuel subassemblies.

Development of a novel Japanese eel myoblast cell line for application in cultured meat production

The present study investigates the isolation, analysis, and characterization of primary cultured cells derived from the muscle tissue of Japanese eel (Anguilla japonica), culminating in establishing a spontaneously immortalized myoblast cell line, JEM1129. We isolated satellite cells from eel muscle tissue to establish a foundation for cultured eel meat production. While initial cell cultures contained myoblasts, continued passaging led to a decline in myoblast characteristics and an increase in fibroblast-like cells. RNA-Seq and RT-qPCR analyses showed significant downregulation of well-established markers for satellite cells and myoblasts, such as pax7a and myoD, over successive passages, highlighting a loss of myoblastic traits. Single-cell cloning was employed to overcome this challenge and maintain myoblast purity, leading to the successful creation of the JEM1129 cell line. These JEM1129 cells demonstrated enhanced expression of myoblast marker genes, exceeding the initial primary culture cell population. The cells showed strong myotube formation, particularly when cultured in a differentiation medium, indicating their robust potential for muscle development. The JEM1129 cell line represents a significant advancement in the cultivation of eel muscle cells, offering a promising avenue for cultured meat production. The findings contribute to a deeper understanding of muscle cell biology and provide valuable insights into using fish-derived myoblasts for cultured meat production.