Critical Process Research Articles

CircRNAs as a novel class of potential diagnostic biomarkers in bipolar disorders.

Bipolar disorder (BD) is a severe mood disorder with uncertain causes and debilitating signs and symptoms. Gene expression is crucial to the pathophysiology of BD and could be influenced by genetic or epigenetic factors, by either direct modification of mRNA templates or by regulation of post-transcriptional translation. Recent evidence has shown that several critical processes in psychiatric diseases, such as neuronal activity or plasticity, synaptic transmission, and neuronal depolarization, have all been linked to circular RNAs (circRNAs). The circRNA profile of neuronal cells, which may be easily ascertained by a liquid biopsy, may shed light on the molecular pathophysiology of psychiatric disorders, including BD. This approach could aid in future development in diagnosis and treatment. In this review, we provide an in-depth understanding of the roles of circRNAs in the pathophysiology of BD and offer new insight into their potential as emerging diagnostic tools and therapeutic targets.

A Review on the Progress of QbD Approach in Nanosystems Optimization: Current Updates and Strategic Applications

: Nanotechnology has made great strides in developing targeted drug delivery systems over the past few decades. These systems have garnered attention for their unique biological properties and ability to deliver drugs in a stable and sustainable manner. Despite these advances, there are still concerns about quality, efficacy, and safety. Many fabrication techniques still need to be refined to address the complex structures and non-standard manufacturing processes that can impact the quality of drug delivery systems. Recently, optimization techniques such as Quality by Design (QbD) have gained popularity in the pharmaceutical industry. QbD is a structured approach that addresses many technological and trait-related issues by providing a deep understanding of the product and its operations. This review examines the current state of QbD in the design of various nano-drug delivery systems, including lipid nanoparticles, lipid carriers, nano micelles, beaded drug delivery systems, nanospheres, cubosomes, and novel cosmeceuticals. Various mathematical models and statistical tests have been used to identify the parameters that influence the physical characteristics of these nanosystems. Critical process attributes such as particle size, yield, and drug entrapment have been studied to assess risk factors during development. However, critical process parameters are often identified through trial and error. This review highlights common material attributes and process parameters that affect the quality of nano-drug delivery systems. Hence, this survey has disclosed the various material attributes and process parameters, quality variables of different nano-drug systems. QbD designs such as Central drug composite, Design of experiment, D-optimal Design, Box-Benkhen Design, and Face center Design in optimizing the nanosystems have also been added. Conclusively, QbD optimization in nano drug delivery systems is expected to be a time-honored strategy in the forthcoming years.

Coaxial nozzle-assisted electrohydrodynamic printing for microscale 3D cell-laden constructs.

Cell printing has found wide applications in biomedical fields due to its unique capability in fabricating living tissue constructs with precise control over cell arrangements. However, it is still challenging to print cell-laden 3D structures simultaneously with high resolution and high cell viability. Here a coaxial nozzle-assisted electrohydrodynamic cell printing strategy was developed to fabricate living 3D cell-laden constructs. Critical process parameters such as feeding rate and stage moving speed were evaluated to achieve smaller hydrogel filaments. The effect of CaCl2 feeding rate on the printing of 3D alginate hydrogel constructs was also investigated. The results indicated that the presented strategy can print 3D hydrogel structures with relatively uniform filament dimension (about 80 μm) and cell distribution. The viability of the encapsulated cells was over 90%. We envision that the coaxial nozzle-assisted electrohydrodynamic printing will become a promising cell printing strategy to advance biomedical innovations.

Statistics as a Social Activity: Attitudes toward Amalgamating Evidence.

Amalgamation of evidence in statistics is conducted in several ways. Within a study, multiple observations are combined by averaging, or as factors in a likelihood or prediction algorithm. In multilevel modeling or Bayesian analysis, population or prior information is combined with data using the weighted averaging derived from probability modeling. In a scientific research project, inferences from data analysis are interpreted in light of mechanistic models and substantive theories. Within a scholarly or applied research community, data and conclusions from separate laboratories are amalgamated through a series of steps, including peer review, meta-analysis, review articles, and replication studies. These issues have been discussed for many years in the philosophy of science and statistics, gaining attention in recent decades first with the renewed popularity of Bayesian inference and then with concerns about the replication crisis in science. In this article, we review the amalgamation of statistical evidence from different perspectives, connecting the foundations of statistics to the social processes of validation, criticism, and consensus building.

Metabolomics Reveal the Chemical Characteristic of Cigar Tobacco Leaves During Air-Curing Process

Air-curing is a critical process in the production of cigar tobacco leaves. This process involves the vital degradation and transformation of macromolecular substances into micromolecular chemical compounds. In this study, three different varieties of tobacco leaves harvested from the Wuzhishan and Danzhou areas of Hainan, China were used as materials. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was employed to investigate the chemical changes before and after air-curing in the metabolites. Metabolic analysis revealed significant stepwise alterations in the tobacco leaf metabolome during the air-curing process. A total of 172 differential metabolites were identified as significant contributors in the biosynthesis pathways of flavonoids, flavone, flavonol, and other secondary metabolites, among the total analyzed 1841 metabolites obtained from the tobacco leaves. Notably, the contents of flavonoid metabolites, including kaempferol, rutin, chlorogenic acid, and quercetin, exhibited a significant decrease throughout the air-curing period. Correlation analysis indicated a strong association between the biosynthesis of flavonoids and the contents of chlorogenic acid and kaempferol, suggesting their role in regulation the formation of yellow and brown colors in tobacco leaves. These findings provide valuable insights for future investigations into the quality, commercial value, and chemical properties of cigar tobacco.

Thermal and Mechanical Investigation of Friction Stir Welding with Disparate Materials AA6061 and AA7075

Background:: The primary objective of this study is to assess the impact of welding conditions on the mechanical properties of friction stir-welded butt joints created from two distinct aluminium alloys, namely, AA6061 and AA7075. Friction stir welding (FSW), known for its innovation and low-energy solid-state bonding technique, was employed in this research. Methods:: FSW experiments were carried out on both AA6061 and AA7075 alloys using a computer numerical control (CNC) machine. The selection and design of the tool geometry were meticulous, with an emphasis on new pin profiles that are nearly flat at the weld contact point. Precisely, four distinct tool geometries were machined from HC-HCr (High carbon, high chromium steel): Circular, Square, Tapered third, and Triangular. Critical process variables that significantly influence weld quality include rotation speed (800 rpm-1400 rpm) and traverse speed (12 to 25 mm/min). These variables were carefully optimized to achieve flawless welds. During the friction stir welding process, the nugget zone undergoes significant deformation, leading to the formation of a new microstructure that substantially impacts the mechanical properties of the joint. Results:: This study comprehensively investigates the thermal and mechanical properties of friction stir welding using aluminium alloys AA6061 and AA7075, considering various tool shapes. Among the four tool shapes employed, two were found to yield higher hardness values (referred to as BH). Notably, the square-shaped tool produced the highest temperature, reaching up to 690ºC, as determined by thermocouple readings. Based on the findings, the optimal FSW parameters for enhancing hardness involve an axial feed and spindle speed of 800 rpm combined with a feed rate of 15 mm/min. These parameters were identified as crucial for achieving the desired mechanical properties in the friction stir-welded joints. Conclusion:: This study presents new developments in FSW technology, which may have patent implications.

Climate Displacement in Pakistan: (under) reported Frame in Media Discourse on Climate Change in Pakistan

Climate disruption has brought irreversible damage to the world. Since global communities are being widely affected by these changes, masses of the populations are facing climate induced displacement. Such displacements involve internal as well as cross border migration. Climate change is marginalized issue in the hand of media in Pakistan, making climate induced displacement doubly underreported. This study focuses on the media coverage of climate change displacement in Pakistani media discourse. The key research objectives include; to find frequency of climate related news during flood coverage in Pakistan, to assess the nature of themes covered while reporting flood related news during defined period of time and to find climate change induced displacement in media coverage of floods in Pakistan. This study uses content analysis as a research method. Time frame focused in the current study involves 15th June 2022 to 15th September 2022, when flood hit major part of Pakistan. Findings of the study reveal that during defined period of time climate change induced displacement remained under reported in Pakistani newspapers (Dawn, The Nation and The Express Tribune). Moreover, while covering irregular weather events pertaining to climate change, main focus remained on damage, government criticism and aid related process.

Quasi-static filling of a disordered nanoporous medium with a non-wetting liquid as a process of self-organized criticality

The paper shows that the quasi-static process of filling a nanoporous medium with a non-wetting liquid is a process of self-organized criticality. In the system, there are distributed stepwise noncorrelated avalanches, separated both in space and time. In addition, it is shown that in the vicinity of the critical point (the threshold pressure of the filling) and above it the “avalanche” clusters of all possible sizes arise. The revealed mechanism allows to pass further to consideration of anomalously slow relaxation as the SOC process and to understanding of uniform internal mechanisms underlying the observed phenomena in such systems.

Comparative analysis of musicality in the works of Kandinsky and Zhu Dequn

The object of the study is the works of the founder of abstract expressionism, Vasily Kandinsky, and the Chinese artist Zhu Dequn. The subject of the study is musicality in the works of Vasily Kandinsky and Zhu Dequn. The author conducts a detailed comparative analysis of abstract ideas and works of two artists in order to study their emotions and sources of inspiration. Special attention is paid to the influence of V. Kandinsky's works on Zhu Dequn's work, as a result of which the Chinese artist tried to combine Western music with the rhythms of Chinese calligraphy, giving his works a deeper meaning. In addition, from the standpoint of a sense of musicality, Zhu Dequn's works such as "The Rhythm of Rebirth" and "Pacification" are considered in the article. It is noted that Zhu Dequn's work is focused on reflecting the spirit, culture and emotions. To achieve the goal set in the research process, approaches and methods of modern philosophy, cultural studies, and art criticism were used. The main conclusions of the study are that under the influence of V. Kandinsky's theory, Zhu Dequn became more musical in his abstract paintings. Despite the fact that Zhu Dequn was not very experienced in music theory and performance, he could accurately capture the melody and rhythm of instruments such as traditional Chinese guzheng, as well as feel the light and color created by a musical composition. The author's main contribution to the study is to demonstrate the influence of V. Kandinsky on the works of Zhu Dequn, who managed to incorporate philosophical ideas and elements of local culture into his works under the influence of music, thereby creating a unique Chinese abstract art. The novelty of the study lies in the fact that the author described how Zhu Dequn's paintings reflected a combination of Chinese and Western painting, which is a valuable example for the exchange and development of Chinese and Western culture and art.

Brain serotonin and serotonin transporter expression in male and female postnatal rat offspring in response to perturbed early life dietary exposures.

Serotonin (5-HT) is critical for neurodevelopment and the serotonin transporter (SERT) modulates serotonin levels. Perturbed prenatal and postnatal dietary exposures affect the developing offspring predisposing to neurobehavioral disorders in the adult. We hypothesized that the postnatal brain 5-HT-SERT imbalance associated with gut dysbiosis forms the contributing gut-brain axis dependent mechanism responsible for such ultimate phenotypes. Employing maternal diet restricted (IUGR, n=8) and high fat+high fructose (HFhf, n=6) dietary modifications, rodent brain serotonin was assessed temporally by ELISA and SERT by quantitative Western blot analysis. Simultaneously, colonic microbiome studies were performed. At early postnatal (P) day 2 no changes in the IUGR, but a ~24% reduction in serotonin (p = 0.00005) in the HFhf group occurred, particularly in the males (p = 0.000007) revealing a male versus female difference (p = 0.006). No such changes in SERT concentrations emerged. At late P21 the IUGR group reared on HFhf (IUGR/HFhf, (n = 4) diet revealed increased serotonin by ~53% in males (p = 0.0001) and 36% in females (p = 0.023). While only females demonstrated a ~40% decrease in serotonin (p = 0.010), the males only trended lower without a significant change within the HFhf group (p = 0.146). SERT on the other hand was no different in HFhf or IUGR/RC, with only the female IUGR/HFhf revealing a 28% decrease (p = 0.036). In colonic microbiome studies, serotonin-producing Bacteriodes increased with decreased Lactobacillus at P2, while the serotonin-producing Streptococcus species increased in IUGR/HFhf at P21. Sex-specific changes emerged in association with brain serotonin or SERT in the case of Alistipase, Anaeroplasma, Blautia, Doria, Lactococcus, Proteus, and Roseburia genera. We conclude that an imbalanced 5-HT-SERT axis during postnatal brain development is sex-specific and induced by maternal dietary modifications related to postnatal gut dysbiosis. We speculate that these early changes albeit transient may permanently alter critical neural maturational processes affecting circuitry formation, thereby perturbing the neuropsychiatric equipoise.

Pre-analytical stability of the CEA, CYFRA 21.1, NSE, CA125 and HE4 tumor markers.

For lung cancer, circulating tumor markers (TM) are available to guide clinical treatment decisions. To ensure adequate accuracy, pre-analytical instabilities need to be known and addressed in the pre-analytical laboratory protocols. This study investigates the pre-analytical stability of CA125, CEA, CYFRA 21.1, HE4 and NSE for the following pre-analytical variables and procedures; i) whole blood stability, ii) serum freeze-thaw cycles, iii) electric vibration mixing and iv) serum storage at different temperatures. Left-over patient samples were used and for every investigated variable six patient samples were used and analysed in duplicate. Acceptance criteria were based on analytical performance specifications based on biological variation and significant differences with baseline. Whole blood was stable for at least 6 hours for all TM except for NSE. Two freeze-thaw cycles were acceptable for all TM except CYFRA 21.1. Electric vibration mixing was allowed for all TM except for CYFRA 21.1. Serum stability at 4°C was 7 days for CEA, CA125, CYFRA 21.1 and HE4 and 4 hours for NSE. Critical pre-analytical processing step conditions were identified that, if not taken into account, will result in reporting of erroneous TM results.

First trimester maternal tryptophan metabolism and embryonic and fetal growth: the Rotterdam Periconceptional Cohort (Predict Study).

What is the association between first trimester maternal tryptophan (TRP) metabolites and embryonic and fetal growth? Higher 5-hydroxytryptophan (5-HTP) concentrations are associated with reduced embryonic growth and fetal growth and with an increased risk of small-for-gestational age (SGA), while higher kynurenine (KYN) concentrations are associated with a reduced risk of SGA. The maternal TRP metabolism is involved in many critical processes for embryonic and fetal growth, including immune modulation and regulation of vascular tone. Disturbances in TRP metabolism are associated with adverse maternal and fetal outcomes. This study was embedded within the Rotterdam Periconceptional Cohort (Predict Study), an ongoing prospective observational cohort conducted at a tertiary hospital from November 2010 onwards. A total of 1115 women were included before 11 weeks of gestation between November 2010 and December 2020. Maternal serum samples were collected between 7 and 11 weeks of gestation, and TRP metabolites (TRP, KYN, 5-HTP, 5-hydroxytryptamine, and 5-hydroxyindoleacetic acid) were determined using a validated liquid chromatography (tandem) mass spectrometry method. Serial 3D ultrasound scans were performed at 7, 9, and 11 weeks of gestation to accurately assess features of embryonic growth, including crown-rump length (CRL) and embryonic volume (EV) offline using virtual reality systems. Fetal growth parameters were retrieved from medical records and standardized according to Dutch reference curves. Mixed models were used to assess associations between maternal TRP metabolites and CRL and EV trajectories. Linear and logistic regression models were utilized to investigate associations with estimated fetal weight (EFW) and birthweight, and with SGA, respectively. All analyses were adjusted for potential confounders. Maternal 5-HTP concentrations and the maternal 5-HTP/TRP ratio were inversely associated with embryonic growth (5-HTP, √CRL: β = -0.015, 95% CI = -0.028 to -0.001; 5-HTP 3√EV: β = -0.009, 95% CI = -0.016 to -0.003). An increased maternal 5-HTP/TRP ratio was also associated with lower EFW and birthweight, and with an increased risk of SGA (odds ratio (OR) = 1.006, 95% CI = 1.00-1.013). In contrast, higher maternal KYN concentrations were associated with a reduced risk of SGA in the unadjusted models (OR = 0.548, 95% CI = 0.320-0.921). Residual confounding cannot be ruled out because of the observational design of this study. Moreover, this study was conducted in a single tertiary hospital, which assures high internal validity but may limit external validity. The novel finding that maternal 5-HTP concentrations are associated with a smaller embryo and fetus implies that disturbances of the maternal serotonin pathway in the first trimester of pregnancy are potentially involved in the pathophysiology of fetal growth restriction. The association between higher maternal KYN concentrations and a reduced risk of SGA substantiate the evidence that the KYN pathway has an important role in fetal growth. More research is needed to delve deeper into the potential role of the maternal TRP metabolism during the periconception period and pregnancy outcome for mother and offspring. This study was funded by the Department of Obstetrics and Gynecology and the Department of Clinical Chemistry of the Erasmus MC, University Medical Center, Rotterdam, the Netherlands. The authors have no competing interests to disclose. N/A.

When will we have a clone? An industry perspective on the typical CLD timeline.

Cell line development (CLD) represents a complex but highly critical process during the development of a biological drug. To shed light on this crucial workflow, a team of BioPhorum members (authors) has developed and executed surveys focused on the activities and effort involved in a typical CLD campaign. An average of 27 members from different companies that participate in the BioPhorum CLD working group answered surveys covering three distinguishable stages of a standard CLD process: (1) Pre-transfection, including vector design and construction; (2) Transfection, spanning the initial introduction of vector into cells and subsequent selection and analysis of the pools; and (3) Single Cell Cloning and Lead Clone Selection, comprising methods of isolating single cells and confirming clonal origin, subsequent expansion and screening processes, and methods for identifying and banking lead clones. The surveys were very extensive, including a total of 341 questions split between antibody and complex molecule CLD processes. In this survey review, the authors interpret and highlight responses for antibody development and, where relevant, contrast complex molecule development challenges to provide a comprehensive industry perspective on the typical time and effort required to develop a CHO production cell line.

New Suns

New Suns

Synthesizing Recent Trends in Interventions and Key Ecosystem Services in Indonesian Peatland

This study conducted a systematic literature review focusing on peatlands studies in Southeast Asia, specifically in Jambi, South Sumatra, and the Riau province of Indonesia, covering the period from 2001 to 2023. To ensure the quality and rigor of the analyzed articles, a critical process and systematic review were employed. Journal articles were extracted using reputable resources, including Google Scholar and Scopus, to enhance the validity and reliability of the research results. We identified significant research topics based on region, province, and sector. Additionally, we synthesized the existing classification of ecosystem services, drawing on previous studies conducted in Indonesia. These services were categorized as provisioning, regulating, cultural, and supporting services. We also reviewed the classification of ecosystem service types based on peatland degradation and restoration. This study identified evidence of peatland intervention to evaluate ecosystem services in Indonesia. We found that large-scale cultivation and production of palm oil, local policies, and forest fires were the main intervening factors in Indonesian peatlands. Furthermore, Indonesian peatlands have undergone conversion to oil palm, timber, and crop plantations. It is imperative to substantiate the effectiveness of future peatland restoration plans and further refine the quantification of services provided by peatland ecosystems through cooperative projects.

Assessment of Benilde Associates towards a Dynamic Business Continuity Model in Disruptive Times

Business continuity and disaster recovery planning are comprised of activities that an organization must perform to resume operations in an unexpected event. The De La Salle College of Saint Benilde SMIT research study adopted the Business Continuity Model and identified gaps that will lead to the proposed Dynamic Business Continuity Model in disruptive times. From this problem, the research question is to what extent BCM factors impact levels (depth and breadth) of the Business Continuity Model during disruptive times. Data was collected by surveying Benilde associates in various academic positions and the Office Center. The data collected was treated using Exploratory Factor Analysis and Multiple Regression for causal relationships. DLS CSB SMIT identified gaps and challenges for faculty and staff in several dimensions in the existing Business Continuity Model factors and the ff are: a. Systems/Data Recovery b. Organizational Support c. Employee Engagement d. Forecasting Capacity and among the identified gaps- “Organizational Support ‘ is considered the most significant among the 4 factors. This consists of Student Support Services, Hunan Resource, and Registrar’s Office. To prevent the interruption in DLS CSB SMIT operations and the survival of an organization during disruptive times, Transparency of the BCM Strategy cascaded to Benilde associates, the importance of creating awareness and being prepared for the unexpected. Business Continuity Model is an essential element that should form part of every critical business process within the College. It is imperative to understand the factors that were identified as gaps that the Institution could be faced with and also understand the steps that should be taken to address and manage these risks. This dissertation paper led to the development of a Dynamic Business Continuity Model which is essential to prepare DLS CSB for any disaster or crisis. The Model serves as a guide for all Centers on what they should do in such an event to ensure that the organizations will survive during the fortuitous event and how to restore the Business and Institution to normalcy. This Research study could provide a Business Continuity Model framework for leaders to assess their organization’s capacity from an unexpected disruption and be used by organizational leaders to assign contingency planning responsibilities or hire business continuity and disaster recovery planning professionals. Keywords: Business Continuity, Disruptive, Dynamic Business Model, Systems/Data Recovery, Organizational Support, Employee Engagement, Forecasting Capacity

The emerging significance of RNA 5-methylcytosine modification in human cancers

Abstract RNA methylation, the most prevalent type of RNA modification, encompasses more than 60 % of all known RNA modifications. With the advancement of methylation sequencing technologies, a diverse range of biological functions associated with RNA methylation in eukaryotes has been revealed. Specifically, 5-methylcytosine (m5C) modifications have been extensively observed in various RNA molecules, including messenger RNAs (mRNAs), ribosomal RNAs (rRNAs), transfer RNAs (tRNAs), and non-coding RNAs. These m5C modifications have been shown to impact critical RNA processes, such as mRNA translation, rRNA assembly, and tRNA stability. Notably, emerging evidence suggests that m5C modifications play significant roles in the initiation and progression of human cancers. However, a comprehensive understanding of the intricate m5C networks involved in human cancers is yet to be fully realized. In this comprehensive review, we provide an up-to-date summary of the roles and potential mechanisms underlying m5C modification in human cancers.

Health management risks in developing countries: Case of a sterilization unit in a Moroccan public hospital

BackgroundThe sterilization of instruments is a critical process to perform the activity in the operating rooms and ensure the safety of patients. The best approach to improve the performance of the sterilization is the risk management. Nevertheless, improving the performance in sterilization units in developing countries presents a real challenge. PurposeDesigning an effective risk management project in health sector, specifically in sterilization units, adaptable to the context of developing countries (DC). MethodsThis study uses Failure Modes and Effects Analysis (FMEA) to manage risks in healthcare in DC. Modifications to the implementation of the recommended tool mode. A case study of the sterilization unit of a tertiary Moroccan national reference university hospital was conducted to verify the effectiveness of the model. FindingsThe present study demonstrated that the proposed model is adaptable to the DC context. This model was implemented with minimal resources and was highly efficient. ConclusionsThe increase of risks in healthcare is a challenge for managers in DC. This study highlights the first successful experience of risk management in Moroccan healthcare sector.

Seed Train Optimization in Microcarrier-Based Cell Culture Post In Situ Cell Detachment through Scale-Down Hybrid Modeling.

Microcarrier-based cell culture is a commonly used method to facilitate the growth of anchorage-dependent cells like MA 104 for antigen manufacturing. However, conventionally, static cell culture is employed for cell propagation before seeding the production bioreactor with microcarriers (MCs). This study demonstrates the effective replacement of the conventional method by serial subculturing on MCs with in situ cell detachment under optimal conditions in closed culture units. This study proves that MA 104 can be subcultured at least five times on Cytodex 1 MC without the need for separating cells and MC after cell harvest. Process parameters impacting cell growth were studied post in situ cell detachment in a scaled-down model. Optimization, using augmented Design of Experiments (DoE) combined with hybrid modeling, facilitated rapid screening of the design space for critical process parameters (CPPs). Optimized conditions included an inoculation density of >16 cells/bead, 3.5-4.5 g/L of Cytodex 1, and a controlled agitation speed, starting at Njs (minimum agitation speed) for the first day with a maximum increase of 25% thereafter. With these design spaces for CPPs, a cell density of 2.6 ± 0.5 × 106 cells/mL was achieved after five days. This refined bioprocess methodology offers a reliable and efficient approach for seed training in stirred tank reactors, which is particularly beneficial for viral vaccine production.

Arresting the bad seed: HDAC3 regulates proliferation of different microglia after ischemic stroke.

The accumulation of self-renewed polarized microglia in the penumbra is a critical neuroinflammatory process after ischemic stroke, leading to secondary demyelination and neuronal loss. Although known to regulate tumor cell proliferation and neuroinflammation, HDAC3's role in microgliosis and microglial polarization remains unclear. We demonstrated that microglial HDAC3 knockout (HDAC3-miKO) ameliorated poststroke long-term functional and histological outcomes. RNA-seq analysis revealed mitosis as the primary process affected in HDAC3-deficent microglia following stroke. Notably, HDAC3-miKO specifically inhibited proliferation of proinflammatory microglia without affecting anti-inflammatory microglia, preventing microglial transition to a proinflammatory state. Moreover, ATAC-seq showed that HDAC3-miKO induced closing of accessible regions enriched with PU.1 motifs. Overexpressing microglial PU.1 via an AAV approach reversed HDAC3-miKO-induced proliferation inhibition and protective effects on ischemic stroke, indicating PU.1 as a downstream molecule that mediates HDAC3's effects on stroke. These findings uncovered that HDAC3/PU.1 axis, which mediated differential proliferation-related reprogramming in different microglia populations, drove poststroke inflammatory state transition, and contributed to pathophysiology of ischemic stroke.