Central Regulator Research Articles

Identifying and costing common gaps in Central and West Africa pharmaceutical regulation.

Regulatory systems strengthening is crucial for catalyzing access to safe and effective medical products and health technologies (MPHT) for all. Identifying and addressing common regulatory gaps through regional approaches could be instrumental for the newly incepted African Medicine Agency. This original study sheds light on common gaps among 10 national regulatory authorities (NRAs) and ways to address them regionally. The study used NRA self-assessment outcomes to identify common gaps in four critical regulatory pillars and estimate the cost of addressing them from regional perspectives that aimed at raising the maturity level of regulatory institutions. A cross-sectional study, using the WHO Global Benchmarking Tool (GBT), was conducted between 2020 and 2021 with five NRAs from ECCAS and ECOWAS member states that use French and Spanish as lingua franca. The 10 NRAs operated in a non-formal-to-reactive approach (ML1-2), which hinders their ability to ensure the quality of MPHT and respond appropriately to public health emergencies. Common gaps were identified in four critical regulatory pillars-good regulatory practices, preparedness for public health emergencies, quality management systems, and substandard and falsified medical products-with overall cost to address gaps estimated at US$3.3 million. We elaborated a reproducible method to strengthen regulatory systems at a regional level to improve equitable access to assured-quality MPHT. Our bottom-up approach could be utilized by RECs to address common gaps through common efforts.

Synchronous Regulation of D-Band Centers in Zn Substrates and Weakening Pauli Repulsion of Zn Ions Using the Ascorbic Acid Additive for Reversible Zinc Anodes.

The advanced aqueous zinc-ion batteries (AZIBs) are still challenging due to the harmful reactions including hydrogen evolution and corrosion. Here, a natural small molecule acid vitamin C (Vc) as an aqueous electrolyte additive has been selectively identified. The small molecule Vc can adjust the d band center of Zn substrate which fixes the active H+ so that the hydrogen evolution reaction (HER) is restrained. Simultaneously, it could also fine-tune the solvation structure of Zn ions due to the enhanced electrostatics and reduced Pauli repulsion verified by energy decomposition analysis (EDA). Hence, the cell retains an ultra-long cycle performance of over 1300 cycles and a superior Coulombic efficiency (CE) of 99.5 %. The prepared full cells display increased rate capability, cycle lifetime, and self-discharge suppression. Our results shed light on the mechanistic principle of electrolyte additives on the performance improvement of ZIBs, which is anticipated to render a new round of studies.

Navigating the digital frontier: a comparative examination of Central Bank Digital Currency (CBDC) and the Quantum Financial System (QFS)

In the global financial landscape, a transformational shift towards digitization is known, and a comparative analysis of two key components is of particular relevance: central bank digital currency (CBDC) and quantum financial system (QFS). This article explores the dynamic relationship between traditional monetary instruments and new digital currencies, considering the transformative impact and regional nuances of CBDC. The article provides a historical overview of the evolution of the development and implementation of CBDC in the world, analyzes the key factors affecting its implementation in different regions, identifies key technological advances that are the disruptive force of QFS, as well as their impact on the transformation of traditional financial transactions. In the context of the development of CBDC and QFS, the legal frameworks that regulate their work, facilitate or hinder their broad integration into existing financial systems, their impact on traditional monetary policy and, accordingly, on economic stability, inflation and interest rates, and the format of the technological infrastructure are determined, which must be ensured for their successful implementation and operation, as well as the main challenges related to this and ways to solve security and privacy problems in their application. The paper combines qualitative and quantitative analysis to provide a comprehensive and validated dataset for a thorough understanding of digital financial paradigms. Qualitative research was conducted based on structured interviews with experts in the development and implementation of CBDC and QFS, as well as reporting and analytical information on central banks, regulators, and other financial institutions. The empirical study is based on the built OLS model, which describes the dependence coefficient using CBDC (the share of the population that uses it) on the traditional interest rate set by the central bank (taking into account the corrections for the possibility of variation of its changes and unobservable factors). The Python programming language with R libraries and the Stata software package are used for modeling. Tests confirmed the model’s statistical significance and high R-squared value (0.756 ) to begin its ability to explain a significant portion of the variability in CBDC adoption rates. The results showed that, on average, per skin unit increase in the traditional interest rate, post-CBDC rate growth is expected to be 1.74%; when the conventional interest rate is zero, the settlement rate after CBDC is 11.14%. The results of this study can be used to predict the dynamics of public perception of digital currencies. In addition, central banks can adjust interest rates to stimulate or moderate the adoption of CBDCs based on broader strategic macroeconomic measures: 1) during economic downturns, higher interest rates can be used to stimulate the adoption of digital currencies as an alternative financial instrument; 2) during periods of economic growth, interest rates may be lowered to mitigate the intended overheating of CBDC adoption.

SPRING is a Dedicated Licensing Factor for SREBP-Specific Activation by S1P.

SREBP transcription factors are central regulators of lipid metabolism. Their proteolytic activation requires ER to the Golgi translocation and subsequent cleavage by site-1-protease (S1P). Produced as a proprotein, S1P undergoes autocatalytic cleavage from its precursor S1PA to mature S1PC form. Here, we report that SPRING (previously C12ORF29) and S1P interact through their ectodomains, and that this facilitates the autocatalytic cleavage of S1PA into its mature S1PC form. Reciprocally, we identified a S1P recognition-motif in SPRING and demonstrate that S1P-mediated cleavage leads to secretion of the SPRING ectodomain in cells, and in liver-specific Spring knockout (LKO) mice transduced with AAV-mSpring. By reconstituting SPRING variants into SPRINGKO cells we show that the SPRING ectodomain supports proteolytic maturation of S1P and SREBP signaling, but that S1P-mediated SPRING cleavage is not essential for these processes. Absence of SPRING modestly diminishes proteolytic maturation of S1PA→C and trafficking of S1PC to the Golgi. However, despite reaching the Golgi in SPRINGKO cells, S1PC fails to rescue SREBP signaling. Remarkably, whereas SREBP signaling was severely attenuated in SPRINGKO cells and LKO mice, that of ATF6, another S1P substrate, was unaffected in these models. Collectively, our study positions SPRING as a dedicated licensing factor for SREBP-specific activation by S1P.

TRIB3 promotes the progression of renal cell carcinoma by upregulating the lipid droplet-associated protein PLIN2

Abnormal lipid metabolism and lipid accumulation are characteristic hallmarks of renal cell carcinoma (RCC). While there is prior evidence closely linking such lipid accumulation within RCC cells and consequent tumorigenesis, the mechanisms underlying this process remain incompletely understood. In this study, a series of bioinformatics analyses were initially performed by screening RCC databases and gene sets, ultimately leading to the identification of TRIB3 as an oncogene that functions as a central regulator of lipid metabolism. TRIB3 overexpression was observed in both RCC patient tumor tissues and cell lines, and this upregulation was correlated with a worse RCC patient prognosis. When TRIB3 was knocked down, this resulted in a reduction in lipid accumulation and the consequent induction of endoplasmic reticulum (ER) stress-related apoptotic cell death. At the molecular level, interactions between TRIB3 and PLIN2 were found to abrogate TEB4-mediated PLIN2 ubiquitination and consequent degradation, thus maintaining higher PLIN2 expression levels. This simultaneously helps facilitate the accumulation of lipids while preserving ER homeostasis, thus driving accelerated RCC tumor progression. This TRIB3-PLIN2 axis thus represents a promising new target for efforts to treat RCC.

A review of the potential involvement of small RNAs in transgenerational abiotic stress memory in plants.

Crop production is increasingly threatened by the escalating weather events and rising temperatures associated with global climate change. Plants have evolved adaptive mechanisms, including stress memory, to cope with abiotic stresses such as heat, drought, and salinity. Stress memory involves priming, where plants remember prior stress exposures, providing enhanced responses to subsequent stress events. Stress memory can manifest as somatic, intergenerational, or transgenerational memory, persisting for different durations. The chromatin, a central regulator of gene expression, undergoes modifications like DNA acetylation, methylation, and histone variations in response to abiotic stress. Histone modifications, such as H3K4me3 and acetylation, play crucial roles in regulating gene expression. Abiotic stresses like drought and salinity are significant challenges to crop production, leading to yield reductions. Plant responses to stress involve strategies like escape, avoidance, and tolerance, each influencing growth stages differently. Soil salinity affects plant growth by disrupting water potential, causing ion toxicity, and inhibiting nutrient uptake. Understanding plant responses to these stresses requires insights into histone-mediated modifications, chromatin remodeling, and the role of small RNAs in stress memory. Histone-mediated modifications, including acetylation and methylation, contribute to epigenetic stress memory, influencing plant adaptation to environmental stressors. Chromatin remodeling play a crucial role in abiotic stress responses, affecting the expression of stress-related genes. Small RNAs; miRNAs and siRNAs, participate in stress memory pathways by guiding DNA methylation and histone modifications. The interplay of these epigenetic mechanisms helps plants adapt to recurring stress events and enhance their resilience. In conclusion, unraveling the epigenetic mechanisms in plant responses to abiotic stresses provides valuable insights for developing resilient agricultural techniques. Understanding how plants utilize stress memory, histone modifications, chromatin remodeling, and small RNAs is crucial for designing strategies to mitigate the impact of climate change on crop production and global food security.

Impact of methimazole-induced hypothyroidism on postnatal swine.

Thyroid hormones regulate metabolic rate, nutrient utilization, growth, and development. Swine are susceptible to thyroid suppression in response to disease or environmental conditions, but the physiological impact of such disruption has not been established. The objective of this study was to evaluate the impact of hypothyroidism induced with the antithyroid medication methimazole (MMI). 10 mg/kg MMI significantly decreased circulating triiodothyronine (T3) for the duration of treatment but had only a transient effect on circulating thyroxine (T4). Thyroid tissue weight was significantly increased by more than 3.5-fold in response to MMI treatment. Histologically, the eosinophilic colloid was largely absent from the thyroid follicle which displayed a disorganized columnar epithelium consistent with goiter. MMI induced hypothyroidism has no effect on growth rate over 28 days. Hepatic expression of genes associated with thyroid metabolism (DIO1, DIO2, and DIO3), lipid utilization (CD36, FASN, and ACACA), apoptosis (TP53, PERP, SIVA1, and SFN) and proliferation (CDK1, CDK2, CDK4, and CDKN1A) were unaffected by treatment. Collectively these results demonstrate that MMI induces mild systemic hypothyroidism and pronounced goiter, indicating a strong homeostatic central regulation within the hypothalamic pituitary thyroid axis. This combined with limited peripheral effects, indicates resilience to hypothyroidism in modern swine.

Peran Pemerintah Dalam Mengatur Tataniaga Garam: Dominansi Political Will atau Economic Interest? Sebuah Tinjauan Actor-Network Theory

Conflicting interests among parties involved in the salt commodity trade create challenges. This study employs qualitative methods to investigate the interactions between human and non-human actors. The primary data used in this study was collected through in-depth interviews with relevant parties. Additionally, the triangulation method utilizes secondary data from statistical sources such as the Central Statistics Agency (BPS), related Ministries, and regulations issued by related Ministries. The results indicate that the government's efforts to regulate salt trading have primarily focused on regulations and policies related to production, distribution, and imports. However, the government has not been extensively involved in regulating market players, which include salt farmers, landowners, middlemen or trade intermediaries, consumers (including the salt processing industry and salt user industries), and salt importers. According to the perspective of actor-network theory, the government plays a significant role in organizing the interaction of actors in the salt trade. However, the existence of competing interest in the salt trade illustrates that the actors involved have systematically gathered in a network based on their economic interests.

The conserved GOLD domain in the Plasmodiophora brassicae effector Pb257 is required for triggering cell death and root swelling

Plasmodiophora brassicae is one of the biggest threats to cruciferous plants and one of the most economically significant diseases worldwide. However, our current understanding of its pathogenic mechanisms remains limited. In this study, we have identified an effector, Pb257, which plays a crucial role in the virulence of P. brassicae. The expression pattern analysis revealed continuous induction of Pb257 during both primary and secondary infections. Ectopic expression of Pb257 strongly triggered cell death in Nicotiana benthamiana and several other plants, which was dependent on SOBIR1 and the salicylic acid pathway but not BAK1, a central molecular regulator, to mediate classical defense pathways. Overexpression of Pb257 increased susceptibility of Brassica rapa to P. brassicae. Silencing of the Pb257-encoding gene led to reduce root swelling. Further investigation showed that the conserved GOLD domain within Pb257 was essential for inducing root enlargement, which is similar to the mild symptoms of clubroot, indicating that it is an important effector for promoting root cell enlargement. GOLD domain-containing family proteins were widely present in the kingdoms of protozoa, fungi, and animalia, and Pb257 was clustered into a clade of protozoa, suggesting the encoding gene as a potential marker gene for classification of P. brassicae. The GOLD domain-containing proteins are known regulators involved in human cell proliferation and immune system disruption, however, no previous reports have described a pathogen-derived GOLD domain protein with elicitor activity. Our findings demonstrate that Pb257 functions as a crucial effector contributing to the virulence of P. brassicae and could be used as a potential molecular target for controlling clubroot disease.

RpoS and the bacterial general stress response.

SUMMARYThe general stress response (GSR) is a widespread strategy developed by bacteria to adapt and respond to their changing environments. The GSR is induced by one or multiple simultaneous stresses, as well as during entry into stationary phase and leads to a global response that protects cells against multiple stresses. The alternative sigma factor RpoS is the central GSR regulator in E. coli and conserved in most γ-proteobacteria. In E. coli, RpoS is induced under conditions of nutrient deprivation and other stresses, primarily via the activation of RpoS translation and inhibition of RpoS proteolysis. This review includes recent advances in our understanding of how stresses lead to RpoS induction and a summary of the recent studies attempting to define RpoS-dependent genes and pathways.

Abstract 602: Targeting CDK9 with a novel potent and selective inhibitor, YX0798, in CAR-T-relapsed mantle cell lymphoma models

Abstract Introduction: Cyclin-dependent kinase (CDK) family members play critical roles in multiple cellular processes. CDK9 is a central regulator at several stages of PolII transcription, including initiation, elongation, and termination. CDK9 dysfunction resulted in transcriptomic reprogramming and promotion of the development of cancers, including lymphoma and leukemia. We and others reported that transcriptomic reprogramming is associated with disease progression and therapeutic resistance in mantle cell lymphoma (MCL). Targeting CDK9 with its inhibitors AZD4573 and enitociclib is safe and effective for treatment in preclinical MCL models, and they are currently under clinical investigation to assess their efficacy and safety. The data demonstrated that CDK9 is a promising therapeutic target. In this study, we developed a novel CDK9 inhibitor, YX0798, and assessed its potency and safety in preclinical MCL models. Method: Cell viability assays were conducted in a time- and dose-dependent manner to assess the in vitro efficacy of YX0798 as well as AZD4573. Apoptosis assays and western blots were used to determine the mechanism of action in MCL cells. Patient-derived xenograft models from primary MCL patient samples were established in immunodeficient mice and used to determine the therapeutic potential YX0798. Results: Next generation sequencing showed that CDK9 was upregulated and cancer hallmarks MYC-TARGETS-v1 and -v2 were highly enriched in CAR-T-relapsed MCL patients compared to CAR-T-naïve patients. We hypothesized that CDK9 overexpression and activity may greatly contribute to the enrichment of MYC-TARGETS-v1 and -v2 in patients with CAR-T resistance, so targeting CDK9 may overcome that resistance. YX0798 was developed to be a potent and selective CDK9 inhibitor. It markedly inhibited cell viability and induced robust apoptosis in MCL cell lines. As expected, it inhibited CDK9 kinase activity and suppressed the expression of short-lived proteins like c-MYC and MCL-1 that are critical for MCL cell survival and growth. YX0798 at 2 mg/kg for continuous 7 days via pump or at 5 mg/kg daily by oral administration potently inhibited the tumor growth of xenografts derived from a patient with therapeutic relapse to the CAR-T therapy in vivo (P<0.0001). This treatment efficacy was found to be similar at that of AZD4573 but at a higher dosage (15+15 mg/kg, 2 hours split, QW, IP). No adverse effects in mice were observed for any of treatments. Conclusion: These data show that our novel CDK9 inhibitor YX0798 is potent and efficacious in treating aggressive MCL models and overcame CAR-T resistance at a lower dosage than AZD4573. Citation Format: Vivian Jiang, Yu Xue, Hong Kim, Joseph McIntosh, Haiying Chen, Tianci Zhang, Yang Liu, Jia Zhou, Michael Wang. Targeting CDK9 with a novel potent and selective inhibitor, YX0798, in CAR-T-relapsed mantle cell lymphoma models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 602.

Abstract 1601: The role of complement in pancreatic cancer cachexia

Abstract Cancer cachexia is a multifactorial condition characterized by skeletal muscle atrophy and dysfunction that impairs longevity and quality of life for the vast majority of cancer patients. Recently, we discovered through unbiased proteomic analyses that the complement system is activated in the skeletal muscle of cachectic pancreatic cancer patients. Therefore, the purpose of this study was to investigate the role of complement in pancreatic cancer cachexia. To accomplish this, we obtained genetically modified, C57Bl6/J congenic mice lacking the central regulator of complement, C3 (C3−/− mice). Wild-type (C57Bl6/J, WT, n=12) and C3−/− mice (n=11) (male, 12 weeks old) received orthotopic injections of mouse pancreatic cancer cells (KPC) into the pancreas, and as non-cancer controls, WT (n=8) and C3−/− (n=4) mice received injections of PBS into the pancreas. Mice were monitored daily and euthanized at IACUC-mandated tumor endpoint (body condition score ≤ 2; range: 14-15 days). In-vivo diaphragm function was assessed immediately prior to euthanasia via M-mode ultrasound. Skeletal muscles (soleus, SOL; tibialis anterior, TA; diaphragm, DIA) were harvested and ex-vivo contractile function was assessed on strips of costal diaphragm. Diaphragmatic excursion and respiratory rate were reduced with KPC tumor burden in WT mice, but preserved in C3−/− mice. Deletion of C3 did not influence the KPC-induced reduction of body mass or KPC tumor growth. Atrophy of limb muscle mass (SOL, TA) and fiber size (TA) during KPC tumor burden was attenuated in C3−/− mice. Maximal specific force of DIA was greater in C3−/− versus WT KPC-bearing mice. KPC tumor burden increased collagen content in the DIA of WT mice, but not C3−/− mice. Diaphragm fiber size was reduced in WT KPC-bearing mice, but preserved in C3−/− mice. KPC tumor burden induced significant increases in immune cell (CD45+) infiltration and fibroadipogenic progenitor cell (PDGFRa+) abundance in the DIA of WT tumor-bearing mice, but to a lesser extent in C3−/− mice. These findings suggest complement activation is causative in pancreatic cancer-induced skeletal muscle wasting and dysfunction. Inhibition of complement during cancer cachexia may possess therapeutic potential for the preservation of skeletal muscle mass and function. Citation Format: Andrew C. D'Lugos, Chandler Callaway, Sarah M. Judge, Andrew R. Judge. The role of complement in pancreatic cancer cachexia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1601.

Abstract 6680: High-fiber diet induces favorable gut microbiota, reduces immunosuppression, and inhibits lung cancer development

Abstract Emerging evidence suggests that resident microbial communities act as a central regulator of host homeostasis. The dynamic structure, composition, and function of gut microbiota can be influenced by numerous factors, among which nutritional profile act as most crucial determinant. Microbial consortia inhabiting the gut have been implicated in the etiology and pathogenesis of various cancers and are known to influence cancer immunosurveillance and response to immunotherapy. Despite these known associations, the underlying mechanisms by which dietary interventions such as those rich in fiber (high-fiber diet) regulate gut-immune axis in prevention of lung cancer are poorly defined. Here, we employed a human-relevant mouse model in which knockout of G-protein coupled receptor 5A (Gprc5a−/−) and exposure to tobacco lead to development of preneoplasia and lung adenocarcinoma (LUAD). Mice were randomly grouped based on dietary treatment: no fiber (control), 10% cellulose, 10% pectin-inulin following weaning (early life) or after transplant of MDA-F471 NNK-induced LUAD cells (dietary intervention). Tumor growth and volumes were most significantly reduced in mice fed a high-fiber pectin-inulin diet. Notably, anti-tumor effects of the high-fiber diet were observed in both early life and later dietary intervention settings. 16S rRNA-Seq analysis of fecal pellets revealed significant differences in gut alpha as well as beta diversity, the latter indicating clear separation of community structure between fiber-deprived versus high-fiber diet groups. Evidently, gut microbiota of pectin-inulin fed mice displayed significantly elevated relative abundance of Bacteroides sartorii, Bacteroides caecimuris, Akkermansia muciniphila and Bifidobacterium pseudolongum, taxa that have been reported to be strongly associated with favorable responses to immunotherapy. High-fiber fed mice also showed significantly reduced fractions of tumor-promoting genera such as Odoribacter and Alistipes. These effects were accompanied by widespread changes in the tumor immune microenvironment between fiber-rich and fiber-poor groups. Pectin-inulin fed mice displayed enhanced antitumor immune responses evident by remarkably increased fractions of B lineage cells, cDC1s and M1-likemacrophages and, conversely, reduced levels of activated cDC2s, M2 macrophages and polymorphonuclear myeloid-derived suppressor cells. Our findings inform on novel mechanisms by which dietary fiber robustly impacts gut consortia and immune milieu in the inhibition of LUAD. Our findings also suggest that the incorporation of dietary fiber in cancer treatment (e.g., immunotherapy) could improve outcomes for patients with LUAD. Citation Format: Manvi Sharma, Yuejiang Liu, Hannah Moreno, Inti C. Reischle, Mohamed A. Jamal, Tieling Zhou, Zahraa Rahal, Chidera O. Chukwuocha, Ke Liang, Robert R. Jenq, Matthew C. Ross, Kristi L. Hoffman, Humam Kadara. High-fiber diet induces favorable gut microbiota, reduces immunosuppression, and inhibits lung cancer development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6680.

P53 regulates diverse tissue-specific outcomes to endogenous DNA damage in mice

DNA repair deficiency can lead to segmental phenotypes in humans and mice, in which certain tissues lose homeostasis while others remain seemingly unaffected. This may be due to different tissues facing varying levels of damage or having different reliance on specific DNA repair pathways. However, we find that the cellular response to DNA damage determines different tissue-specific outcomes. Here, we use a mouse model of the human XPF-ERCC1 progeroid syndrome (XFE) caused by loss of DNA repair. We find that p53, a central regulator of the cellular response to DNA damage, regulates tissue dysfunction in Ercc1-/- mice in different ways. We show that ablation of p53 rescues the loss of hematopoietic stem cells, and has no effect on kidney, germ cell or brain dysfunction, but exacerbates liver pathology and polyploidisation. Mechanistically, we find that p53 ablation led to the loss of cell-cycle regulation in the liver, with reduced p21 expression. Eventually, p16/Cdkn2a expression is induced, serving as a fail-safe brake to proliferation in the absence of the p53-p21 axis. Taken together, our data show that distinct and tissue-specific functions of p53, in response to DNA damage, play a crucial role in regulating tissue-specific phenotypes.

CONSTANS, a HUB for all seasons: How photoperiod pervades plant physiology regulatory circuits.

How does a plant detect the changing seasons and make important developmental decisions accordingly? How do they incorporate daylength information into their routine physiological processes? Photoperiodism, or the capacity to measure the daylength, is a crucial aspect of plant development that helps plants determine the best time of the year to make vital decisions, such as flowering. The protein CONSTANS (CO) constitutes the central regulator of this sensing mechanism, not only activating florigen production in the leaves but also participating in many physiological aspects in which seasonality is important. Recent discoveries place CO in the center of a gene network that can determine the length of the day and confer seasonal input to aspects of plant development and physiology as important as senescence, seed size, or circadian rhythms. In this review, we discuss the importance of CO protein structure, function, and evolutionary mechanisms that embryophytes have developed to incorporate annual information into their physiology.

Systemic and strict regulation of the glutathione redox state in mitochondria and cytosol is needed for zebrafish ontogeny

BackgroundRedox control seems to be indispensable for proper embryonic development. The ratio between glutathione (GSH) and its oxidized disulfide (GSSG) is the most abundant cellular redox circuit. MethodsWe used zebrafish harboring the glutaredoxin 1-redox sensitive green fluorescent protein (Grx1-roGFP) probe either in mitochondria or cytosol to test the hypothesis that the GSH:GSSG ratio is strictly regulated through zebrafish embryogenesis to sustain the different developmental processes of the embryo. ResultsFollowing the GSSG:GSH ratio as a proxy for the GSH-dependent reduction potential (EhGSH) revealed increasing mitochondrial and cytosolic EhGSH during cleavage and gastrulation. During organogenesis, cytosolic EhGSH decreased, while that of mitochondria remained high. The similarity between EhGSH in brain and muscle suggests a central regulation. Modulation of GSH metabolism had only modest effects on the GSSG:GSH ratios of newly hatched larvae. However, inhibition of GSH reductase directly after fertilization led to dead embryos already 10 h later. Exposure to the emerging environmental pollutant Perfluorooctane Sulfonate (PFOS) disturbed the apparent regulated EhGSH as well. ConclusionsMitochondrial and cytosolic GSSG:GSH ratios are almost identical in different organs during zebrafish development indicating that the EhGSH might follow H2O2 levels and rather indirectly affect specific enzymatic activities needed for proper embryogenesis. General significanceOur data confirm that vertebrate embryogenesis depends on strictly regulated redox homeostasis. Disturbance of the GSSG:GSH circuit, e.g. induced by environmental pollution, leads to malformation and death.

The AMPK-related kinase NUAK1 controls cortical axons branching by locally modulating mitochondrial metabolic functions

The cellular mechanisms underlying axonal morphogenesis are essential to the formation of functional neuronal networks. We previously identified the autism-linked kinase NUAK1 as a central regulator of axon branching through the control of mitochondria trafficking. However, (1) the relationship between mitochondrial position, function and axon branching and (2) the downstream effectors whereby NUAK1 regulates axon branching remain unknown. Here, we report that mitochondria recruitment to synaptic boutons supports collateral branches stabilization rather than formation in mouse cortical neurons. NUAK1 deficiency significantly impairs mitochondrial metabolism and axonal ATP concentration, and upregulation of mitochondrial function is sufficient to rescue axonal branching in NUAK1 null neurons in vitro and in vivo. Finally, we found that NUAK1 regulates axon branching through the mitochondria-targeted microprotein BRAWNIN. Our results demonstrate that NUAK1 exerts a dual function during axon branching through its ability to control mitochondrial distribution and metabolic activity.

Strategies of Regional Levies of the Marine Affairs and Fisheries Office of Central Java Province of Indonesia

There is still a need to properly increase the role of regional levies in generating revenue for Central Java Province. This is because each region has been granted the authority to manage its affairs, including regional finance, with the expectation that regional governments will become financially independent from the central government. In reality, many regions in Indonesia and other countries still significantly rely on the central government for financial support. Therefore, regional governments must focus on increasing revenue by improving the management of regional levies at the Regional Apparatus Organizations. This is achieved through the development of strategies for the regional levies sector based on the SWOT analysis results and the projected regional levies revenue of the Marine Affairs and Fisheries Office of Central Java Province. This research aimed to identify strategies, create strategies for regional levies, and analyze the projected revenue from these levies. This is expected to support efforts to increase regional levies revenue. This research used primary and secondary data. The primary data were obtained through interview results with key informants to assess the strengths, weaknesses, opportunities, and threats faced by the Marine Affairs and Fisheries Office in collecting regional levies. Meanwhile, secondary data comprised information on regional levies and relevant references on the Regional Apparatus Organization. The data collected were analyzed using the SWOT and multiple linear regression analyses as analytical techniques. The results showed the strengths, weaknesses, opportunities, and threats used to collect regional levies and also the strategies. Based on the projected revenue, the levies generated by this Regional Apparatus Organization are projected to increase from 2022 to 2027 steadily. The implications include recommendations for activities carried out by the Marine Affairs and Fisheries Office, which aligned with the potential and capabilities of the regional and central government regulations governing regional levies.

Dietary Iron Overload Triggers Hepatic Metabolic Disorders and Inflammation in Laying Hen.

Iron, an essential trace element, is involved in various physiological processes; however, consumption of excessive iron possesses detrimental effects. In practical feed production, the iron content added to feeds often far exceeds the actual demand, resulting in an excess of iron in the body. The liver as a central regulator of iron homeostasis is susceptible to damage caused by disorders in iron metabolism. A model of hepatic iron overload in laying hens was developed in this study by incorporating iron into their diet, and the specific mechanisms underlying iron overload-induced hepatic injury were investigated. Firstly, this study revealed that a high-iron diet resulted in hepatic iron overload, accompanied by impaired liver function. Next, assessment of oxidative stress markers indicated a decrease in activities of T-SOD and CAT, coupled with an increase in MDA content, pointing to the iron-overloaded liver oxidative stress. Thirdly, the impact of iron overload on hepatic glycolipid and bile acid metabolism-related gene expressions were explored, including PPAR-α, GLUT2, and CYP7A1, highlighting disruptions in hepatic metabolism. Subsequently, analyses of inflammation-related genes such as iNOS and IL-1β at both protein and mRNA levels demonstrated the presence of inflammation in the liver under conditions of dietary iron overload. Overall, this study provided comprehensive evidence that dietary iron overload contributed to disorders in glycolipid and bile acid metabolism, accompanied by inflammatory responses in laying hens. Further detailing the specific pathways involved and the implications of these findings could offer valuable insights for future research and practical applications in poultry nutrition.

The NADase CD38 is a central regulator in gouty inflammation and a novel druggable therapeutic target

ObjectivesCellular NAD+ declines in inflammatory states associated with increased activity of the leukocyte-expressed NADase CD38. In this study, we tested the potential role of therapeutically targeting CD38 and NAD+ in gout.MethodsWe studied cultured mouse wild type and CD38 knockout (KO) murine bone marrow derived macrophages (BMDMs) stimulated by monosodium urate (MSU) crystals and used the air pouch gouty inflammation model.ResultsMSU crystals induced CD38 in BMDMs in vitro, associated with NAD+ depletion, and IL-1β and CXCL1 release, effects reversed by pharmacologic CD38 inhibitors (apigenin, 78c). Mouse air pouch inflammatory responses to MSU crystals were blunted by CD38 KO and apigenin. Pharmacologic CD38 inhibition suppressed MSU crystal-induced NLRP3 inflammasome activation and increased anti-inflammatory SIRT3–SOD2 activity in macrophages. BMDM RNA-seq analysis of differentially expressed genes (DEGs) revealed CD38 to control multiple MSU crystal-modulated inflammation pathways. Top DEGs included the circadian rhythm modulator GRP176, and the metalloreductase STEAP4 that mediates iron homeostasis, and promotes oxidative stress and NF-κB activation when it is overexpressed.ConclusionsCD38 and NAD+ depletion are druggable targets controlling the MSU crystal- induced inflammation program. Targeting CD38 and NAD+ are potentially novel selective molecular approaches to limit gouty arthritis.