Ayurvedic herbal formulations Haridra Khanda and Manjisthadi Kwath (brihat) in the management of allergic rhinitis: A pharmacological study.

This study aims to pharmacologically validate Haridra Khanda (HK) and Manjishthadi Kwatham (brihat) (MMK) in allergy management using invivo and invitro studies to rationalize the prescription of these two ayurvedic polyherbal drug formulations, which are currently used in Indian government hospitals. Experimental animals received HK and MMK orally from day 0 to day 14 and histamine (1 mg/kg b.w/i.v) and 1 % evans blue (EB) (0.1 mL) via tail vein on day 14. The compound 48/80 (intracutaneous) challenged mice model followed the same technique. The former mimicked acute anaphylaxis and the latter mast cell degranulation. For both models, EB dye leakage was quantified spectrophotometrically to determine vascular permeability. Plasma histamine was measured in Compound 48/80-induced animals using LC-ESI-MS/MS. The guineapig received HK and MMK p.o. and 0.6 % histamine sprayed in a histamine chamber to simulate allergic rhinitis. Blood eosinophil count and sneeze rate were measured in histamine-challenged guineapigs. Goat R.B.C. membrane stability assay (mammalian cell membrane toxicity) and intracellular histamine-induced cytosolic Ca2+ release assay in Chinese hamster ovary (CHO) cells were performed in vitro. For both histamine and Compound 48/80 challenged animals, HK (22.81 % and 14.58 %) and MMK (19.71 % and 22.40 %) significantly reduced EB dye leakage (p < 0.05). Both formulations, HK and MMK considerably (p < 0.05) decreased plasma histamine (29.62 % and 25.37 % respectively) in mice and eosinophilic count (11.56 % and 9.94 % respectively) and sneeze rate (42.58 % and 29.03 % respectively) in guinea pigs. In membrane stability experiment, HK and MMK reduced RBC lysis. Both HK and MMK raw/dialysate blocked CHO cell cytosolic Ca2+ release. HK and MMK activities mimic mast cell stabilization with possible H1 receptor inactivation seen by decreased Ca2+ efflux and thus indicate potential for allergic rhinitis management. The combination of activities is usually related with curative and prophylactic therapy and might lead future clinical trials and therapies.

Child with Closed Head Injury and Persistent Vomiting.

Case Presentation: We present the case of a six-year-old child with autism who presented with persistent vomiting in the setting of a closed head injury (CHI). Computed tomography of the head was normal, but due to persistent vomiting a radiograph of the abdomen was done, which showed multiple, rare-earth magnets in the abdomen. There was no history of witnessed ingestion. These magnets had caused enteroenteric fistula formation leading to persistent vomiting. Discussion: In the setting of CHI, vomiting can be a sign of concussion or intracranial hemorrhage. In cases of CHI where intracranial pathology is ruled out and vomiting still persists, it is important to explore intra-abdominal causes of vomiting, especially in developmentally challenged children as they have higher incidence of unwitnessed foreign body ingestions.

Tran slation , cultural adap tation an d psycho metric anal ysis of U rd u ver sion of Vic torian; institu te of sp or ts ass es sme nt: Achill es quest i onnaire fo r athletes with achilles tendinopathy.

OBJECTIVE: To translate, culturally adapt and psychometrically analyse the Urdu version of the Victorian Institute of Spor t Assessment-Achilles questionnaire. METHODS: The cross-sec tional study was conducted at the Pak istan Sports Board, Lahore, Pakistan, from June 17, 2021, to February 15, 2022, and comprised patients with Achilles tendinopathy in group A and healthy controls in group B. Beaton's guidelines for cultural adaptation and validation for self-repor ted measures were followed to translate and validate the Victorian Institute of Sport Assessment-Achilles questionnaire in Urdu language. Data was analysed using SPSS 23. RESULTS: Of the 180 subjects with mean age 28.06±5.95 years, 125(69.6%) were males. There were 130(72.2%) patients in group A and 50(27.8%) controls in group B. The overall mean score of the Victorian Institute of Sport Assessment- Achille s questionnaire was 55.99±25.43; group A 41.14±9.54 and group B 94.60±4.22. The Urdu version exhibited excellent internal consistency with Cronbach's alpha values 0.95, and excellent test-retest reliability (p<0.001). Absolute reliability was expressed by standard error of measurement 5.317 and minimal detectable change (6.38). Convergent validity demonstrated strong correlation with the physical domain (r=0.81) of the Urdu version of the World Health Organisation Quality of Life Brief Version. CONCLUSIONS: The Victorian Institute of Sport Assessment-Achilles questionnaire could be utilised for assessing severity of Achilles tendinopathy among Urdu-speaking population for clinical as well as research purposes.

Revision Lumbar Spine Surgeries: An Early Career Neurosurgery Experience.

BACKGROUND: The aims and ambitions of a surgeon in the early years of his professional career are to make a good reputation by providing excellent patient outcomes and avoiding complex and difficult surgeries. Revision lumbar spine surgeries (RLSSs) pose a significant challenge in terms of surgical management, as the moribund anatomy increases the risk of complications, adding to an unlikely outcome. OBJECTIVE: We conducted this study to determine the clinical indications and outcomes of RLSSs performed by an early career neurosurgeon. MATERIALS AND METHODS: This cross-sectional study was conducted after approval from the hospital's ethical committee, and data was collected in late December of 2022 and early January 2023, from retrospective records for a single early career neurosurgeon. A form was filled with each patient's data, such as age, gender, time since surgery, indication for surgery, operative findings, types of surgery performed, etc. All variables were noted for the patient and were further categorized, based on the clinical records, into many sub-categories. RESULTS: Almost 400 lumbar spine surgeries were performed by the surgeon, and about 45 (11.25%) were revision surgeries, and the full record was available for 42 surgeries. These patients' ages ranged from 22 to 70 years, and the mean age was about 46.74±13.29 SD. The common symptoms leading to revision surgeries were numbness and pain in 17 (40.5%) patients each; common per-operative findings were recurrent disc in eight patients (19%), infection in nine patients (21.4%), and fibrosis/adhesions in 16 (38.1%); most common surgeries performed were diskectomy in 11 (26.2%) and diskectomy plus release of adhesions in 12 (28.6%); complications occurred in 14 (33%), and good to excellent outcomes was recorded in 29 (69%) cases.  Conclusion: RLSSs are difficult compared to first-time lumbar spine surgeries, and the moribund anatomy predisposes to complications, and better shall be dealt with great care and, at the minimum, shall be embarked upon as a team.

A Case Report and Literature Review of Pheochromocytoma Without Tachycardia.

This case report highlights a 47-year-old woman with an adrenal incidentaloma and a history of polysubstance abuse, finally diagnosed with pheochromocytoma. Characterized by episodic hypertension, headaches, and palpitations, pheochromocytoma is a rare condition with potential complications like uncontrolled hypertension and heart failure. Remarkably, during her 28-day hospitalization, continuous monitoring revealed no instances of tachycardia or arrhythmias despite multiple symptomatic episodes. This finding aligns with reports that while 50-70% of symptomatic pheochromocytoma patients experience palpitations, only about 20% exhibit detectable tachycardia or arrhythmias. This discrepancy suggests varied individual cardiovascular responses to catecholamine surges, possibly due to differences in catecholamine inactivation rates and receptor sensitivity. This case underscores the complexity of pheochromocytoma symptoms and highlights the need for personalized diagnostic and management strategies. Furthermore, it points to a significant gap in understanding the correlation between palpitations and arrhythmia in pheochromocytoma, indicating a critical area for future research.

Blockchain-enabled infrastructural security solution for serverless consortium fog and edge computing.

The robust development of the blockchain distributed ledger, the Internet of Things (IoT), and fog computing-enabled connected devices and nodes has changed our lifestyle nowadays. Due to this, the increased rate of device sales and utilization increases the demand for edge computing technology with collaborative procedures. However, there is a well-established paradigm designed to optimize various distinct quality-of-service requirements, including bandwidth, latency, transmission power, delay, duty cycle, throughput, response, and edge sense, and bring computation and data storage closer to the devices and edges, along with ledger security and privacy during transmission. In this article, we present a systematic review of blockchain Hyperledger enabling fog and edge computing, which integrates as an outsourcing computation over the serverless consortium network environment. The main objective of this article is to classify recently published articles and survey reports on the current status in the domain of edge distributed computing and outsourcing computation, such as fog and edge. In addition, we proposed a blockchain-Hyperledger Sawtooth-enabled serverless edge-based distributed outsourcing computation architecture. This theoretical architecture-based solution delivers robust data security in terms of integrity, transparency, provenance, and privacy-protected preservation in the immutable storage to store the outsourcing computational ledgers. This article also highlights the changes between the proposed taxonomy and the current system based on distinct parameters, such as system security and privacy. Finally, a few open research issues and limitations with promising future directions are listed for future research work.

Identification and characterization of PAL genes involved in the regulation of stem development in Saccharum spontaneum L.

BACKGROUND: Saccharum spontaneum L. is a closely related species of sugarcane and has become an important genetic component of modern sugarcane cultivars. Stem development is one of the important factors for affecting the yield, while the molecular mechanism of stem development remains poorly understanding in S. spontaneum. Phenylalanine ammonia-lyase (PAL) is a vital component of both primary and secondary metabolism, contributing significantly to plant growth, development and stress defense. However, the current knowledge about PAL genes in S. spontaneum is still limited. Thus, identification and characterization of the PAL genes by transcriptome analysis will provide a theoretical basis for further investigation of the function of PAL gene in sugarcane. RESULTS: In this study, 42 of PAL genes were identified, including 26 SsPAL genes from S. spontaneum, 8 ShPAL genes from sugarcane cultivar R570, and 8 SbPAL genes from sorghum. Phylogenetic analysis showed that SsPAL genes were divided into three groups, potentially influenced by long-term natural selection. Notably, 20 SsPAL genes were existed on chromosomes 4 and 5, indicating that they are highly conserved in S. spontaneum. This conservation is likely a result of the prevalence of whole-genome replications within this gene family. The upstream sequence of PAL genes were found to contain conserved cis-acting elements such as G-box and SP1, GT1-motif and CAT-box, which collectively regulate the growth and development of S. spontaneum. Furthermore, quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that SsPAL genes of stem had a significantly upregulated than that of leaves, suggesting that they may promote the stem growth and development, particularly in the + 6 stem (The sixth cane stalk from the top to down) during the growth stage. CONCLUSIONS: The results of this study revealed the molecular characteristics of SsPAL genes and indicated that they may play a vital role in stem growth and development of S. spontaneum. Altogether, our findings will promote the understanding of the molecular mechanism of S. spontaneum stem development, and also contribute to the sugarcane genetic improving.

Cellular therapies in older adults with hematological malignancies: A case-based, state-of-the-art review.

Cellular therapies, including autologous stem cell transplant (ASCT), allogeneic hematopoietic cell transplantation (alloHCT), and chimeric antigen receptor- (CAR-) T cell therapies are essential treatment modalities for many hematological malignancies. Although their use in older adults has substantially increased within the past decades, cellular therapies represent intensive treatment approaches that exclude a large percentage of older adults due to comorbidities and frailty. Under- and overtreatment in older adults with hematologic malignancy is a challenge and many treatment decisions are influenced by chronologic age. The advent of efficient and well-tolerated newer treatment approaches for multiple myeloma has challenged the role of ASCT. In the modern era, there are no randomized clinical trials of transplant versus non-transplant strategies for patients ≥65 years. Nonetheless, ASCT is feasible for selected older patients and does not result in long-term compromise in quality of life. AlloHCT is the only curative approach for acute myeloid leukemia of intermediate and unfavourable risk but carries a significant risk for non-relapse mortality depending on comorbidities, general fitness, and transplant-specific characteristics, such as intensity of conditioning and donor choice. However, alloHCT is feasible in appropriately-selected older adults. Early referral for evaluation is strongly encouraged as this is the most obvious barrier. CAR-T cell therapies have shown unprecedented clinical efficacy and durability in relapsed and refractory diffuse large B cell lymphoma. Its use is well tolerated in older adults, although evidence comes from limited case numbers. Whether patients who are deemed unfit for ASCT qualify for CAR-T cell therapy remains elusive, but the tolerability and efficacy of CAR-T cell therapy appears promising, especially for older patients. The evidence from randomized trials is strong in favor of using a comprehensive geriatric assessment (CGA) to reduce treatment-related toxicities and guide treatment intensity in the care for solid tumors; its use for evaluation of cellular therapies is less evidence-based. However, CGA can provide useful information on patients' fitness, resilient mechanisms, and reveal potential optimization strategies for compensating for vulnerabilities. In this narrative review, we will discuss key questions on cellular therapies in older adults based on illustrative patient cases.

Navigating the Diagnosis and Treatment of Astroblastoma: A Pediatric Case Report.

Astroblastoma, a rare glial tumor of the central nervous system, presents diagnostic and therapeutic challenges due to its low incidence and variable clinical presentations. In this case study, we present the case of an 11-year-old boy with high-grade astroblastoma, highlighting the complexities in diagnosis and treatment. The clinical presentation initially involved right-sided motor weakness, which, after undergoing a brain MRI, revealed a large solid cystic mass in the left parietal lobe. Histopathological examination after undergoing surgery confirmed an astroblastoma with high-grade features, characterized by increased cellularity and high mitotic activity. Immunostaining patterns supported the glial origin of the tumor. Gross total resection remains the primary approach for its treatment, but adjuvant therapies for high-grade astroblastomas are still evolving, offering potential life-changing possibilities for the future. Due to its rarity, collecting sufficient data to develop an effective treatment protocol for this uncommon tumor is very challenging. This case underscores the importance of combined efforts and ongoing research to effectively navigate the diagnosis and treatment of astroblastoma.

Generating human bone marrow organoids for disease modeling and drug discovery.

The bone marrow supports and regulates hematopoiesis, responding to physiological requirements for blood cell production over ontogeny and during pathological challenges. Interactions between hematopoietic cells and niche components are challenging to study mechanistically in the human context, but are important to delineate in order to explore the pathobiology of blood and bone marrow disorders. Organoids are proving transformative in many research settings, but an accurate human bone marrow model incorporating multiple hematopoietic and stromal elements has been lacking. This protocol describes a method to generate three-dimensional, multilineage bone marrow organoids from human induced pluripotent stem cells (hiPSCs), detailing the steps for the directed differentiation of hiPSCs using a series of cytokine cocktails and hydrogel embedding. Over 18 days of differentiation, hiPSCs yield the key lineages that are present in central myelopoietic bone marrow, organized in a well-vascularized architecture that resembles native hematopoietic tissues. This presents a robust, in vitro system that can model healthy and perturbed hematopoiesis in a scalable three-dimensional microenvironment. Bone marrow organoids also support the growth of immortalized cell lines and primary cells from healthy donors and patients with myeloid and lymphoid cancers, including cell types that are poorly viable in standard culture systems. Moreover, we discuss assays for the characterization of organoids, including interrogation of pathogenic remodeling using recombinant TGF-ß treatment, and methods for organoid engraftment with exogenous cells. This protocol can be readily adapted to specific experimental requirements, can be easily implemented by users with tissue culture experience and does not require access to specialist equipment.

Analysis of Photosynthetic Characteristics and Screening High Light-Efficiency Germplasm in Sugarcane.

Sugarcane is a globally significant crop for sugar and energy production, and developing high light-efficiency sugarcane varieties is crucial for enhancing yield and quality. However, limited research is available on the screening of sugarcane germplasm with high photosynthetic efficiency, especially with different leaf positions. The present study, conducted in Guangxi, China, aimed to analyze the photosynthetic characteristics of 258 sugarcane varieties at different leaf positions over three consecutive years in field experiments. The results showed significant differences in photosynthetic characteristics among genotypes, years, and leaf positions. Heritability estimates for various photosynthetic parameters ranged from 0.76 to 0.88. Principal component analysis revealed that the first three principal components accounted for over 99% of the cumulative variance. The first component represented photosynthetic efficiency and light utilization, the second focused on electron transfer and reaction center status, and the third was associated with chlorophyll content. Cluster and discriminant analysis classified sugarcane genotypes into three categories: high photosynthetic efficiency (HPE) with 86 genotypes, medium photosynthetic efficiency (MPE) with 60 genotypes, and low photosynthetic efficiency (LPE) with 112 genotypes. Multi-year trials confirmed that HPE sugarcane genotypes had higher single-stem weight and sucrose content. This study provides valuable insights into the photosynthetic physiological characteristics of different sugarcane varieties, which can contribute to further research regarding high yields and sugar breeding.

Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Scaphium Affine Seed Extract and Assessment of its Anti-Hemorrhoidal Efficacy.

BACKGROUND: Seeds of plant Scaphium Affine are traditionally used by the healers of "India" for the treatment of piles. OBJECTIVE: The primary objective of the study was to assess the anti-hemorrhoidal potential of the ethanolic seed extract of Scaphium affine. METHODS: After the soxhlet extraction method, the seed extract from Scaphium affine was first submitted to phytochemical standardization and then GC-MS analysis. Rats were given Croton oil and Jatropha oil to develop hemorrhoids, and Scaphium affine seed extract (ESA) was administered orally for 5 days and 3 days, respectively, at doses of 1000 and 500 mg/kg. The Rectoanal coefficient (RAC) was calculated as an inflammatory marker. The hemorrhoidal tissues were also subjected to cytokine profiling, biochemical estimation and histopathology. RESULTS: ESA demonstrated the presence of flavonoids, saponins, phytosterols, phenols, and tannins. GCMS analysis elucidated the presence of hexadecanoic acid 2 hydroxy -1,3 propane diyl ester,9 Octadecanoic acid ethyl ester, Cyclohexane 1,4 di methyl cis, Farnesol isomer,1, E-11, Z-13 octa decatriene, Stigmasterol, N-(5 ethyl -1,3,4-thiadiazol-yl) benzamide, N, N Dinitro 1,3,5,7 tetraza bicyclo 93,3,1) as major phytoconstituents. The results depicted more potent anti-hemorrhoidal activity of ESA at 1000 mg/kg, p.o., which was evident through a decrease in RAC. A significant decline in the levels of IL-1ß, IL-6, and TNF-α expression was observed, along with the restoration of altered antioxidants and enzymes. Histopathological analysis confirmed the tissue recovery as it revealed minimal inflammation and decreased dilated blood vessels in treated animals. CONCLUSION: Based on the results it can be concluded that seeds of Scaphium affine showed significant anti-hemorrhoid agents which may be attributed to their anti-inflammatory and anti-oxidant potential due to the presence of certain phytoconstituents in it. The study also supports the traditional use of seeds of Scaphium affine for the first time in the treatment of hemorrhoids.

Fungal community dynamics associated with the outbreaks of sugarcane root rot disease.

Sugarcane is a critical sugar and bioenergy crop in China. However, numerous factors, including root rot disease, hamper its yield. Root rot disease is a severe agricultural issue, reducing yield and threatening sustainable crop production. The current study aimed to explore the fungal community structure, identify and characterize the primary pathogen for sugarcane root rot in Guangzhou, China. Eighty-nine samples of sugarcane root, stalk, rhizosphere soil, and irrigation water were collected from five sites in Guangzhou, China. Subsequently, 276 fungal strains were isolated to identify the primary pathogens. The five most common genera identified were Penicillium, Fusarium, Gongronella, Trichoderma, and Cladosporium. Fusarium was more prevalent in the infected soil samples than in healthy ones. Pathogenic assays of the strains revealed that the strain GX4-46 caused 80% of the disease. The strain was confirmed as Fusarium commune through phylogenetic and genome sequence analysis. Rhizosphere soil samples from different regional crops were collected to better understand the fungal community structure and the primary pathogen. We observed a significant presence of Fusarium in irrigation water, indicating that the root rot disease could originate from the irrigation water and then spread as a soil-borne disease. This research is pioneering and one of the most comprehensive investigations on the occurrence and prevalence of sugarcane root rot disease. This study will serve as a reference for expanding the sugarcane industry and a foundation for further exploration and control of root rot.IMPORTANCESugarcane, a significant economic crop, faces challenges due to root rot pathogens that accumulate each year in plants and soil through ratoon planting. This disrupts soil microbial balance and greatly impedes sugarcane industry growth. Symptoms range from wilting and yellowing leaves to stunted growth and reduced seedling tillers. The rhizosphere microbiota plays an important role in plant development and soil health. Little is known about root rot fungal community structure, especially in sugarcane. Here, we focused on exploring the main causative pathogen of root rot in the area alongside a detailed survey of the rhizosphere soil of different severity sugarcane cultivars and rotation crops of the region. To validate the findings, we also investigated the irrigation water of the area. Our study revealed Fusarium commune as the causative pathogen of root rot in the area, primarily originating from water and later as soil-borne. Using Trichoderma can control the disease effectively.

ASMF: Ambient social media forensics chain of custody with an intelligent digital investigation process using federated learning.

Ambient Intelligence is a concept that relates to a new paradigm of pervasive computing and has the objective of automating responses from the system to humans without any human intervention. In social media forensics, gathering, analyzing, storing, and validating relevant evidence for investigation in a heterogeneous environment is still questionable. There is no hierarchy for automation, even though standardization and secure processes from data collection to validation have not yet been discussed. This poses serious issues for the current investigation procedures and future evidence chain of custody management. This paper contributes threefold. First, it proposes a framework using a blockchain network with a dual chain of data transmission for privacy protection, such as on-chain and off-chain. Second, a protocol is designed to detect and separate local and global cyber threats and undermine multiple federated principles to personalize search space broadly. Third, this study manages personalized updates by means of optimizing backtracking parameters and automating replacements, which directly affects the reduction of negative influence on the social networking environment in terms of imbalanced and distributed data issues. This proposed framework enhances stability in digital investigation. In addition, the simulation uses an extensive social media dataset in different cyberspaces with a variety of cyber threats to investigate. The proposed work outperformed as compared to traditional single-level personalized search and other state-of-the-art schemes.

Can my surgeon scope? Trends in endoscopy training volume and experience among general surgery residents in the United States: a nationwide analysis.

INTRODUCTION: Endoscopy is a major part of surgical training. Accreditation Council for Graduate Medical Education (ACGME) has set standards regarding the minimum volume of endoscopy cases required for graduation. However, there is paucity of high-quality data on the number of cases that most surgical graduates perform. METHODS: We conducted a retrospective analysis of operative case logs of all general surgery residents graduating from ACGME-accredited programs from 2010 to 2023. Data on mean number of endoscopy cases, including mean in each subcategory, were retrieved. Mann-Kendall trend test was used to investigate trends in endoscopy experience. RESULTS: Between 2010 and 2023, the mean overall endoscopy procedures per resident remained stable, with 129.5 in 2010 and 132.1 in 2023 (t = 0.429; p-value = 0.037). The majority of these cases were performed as surgeon junior (76.6% in 2010; 80.9% in 2023), while the remaining cases were logged as surgeon chief. The most substantial contribution to the overall volume was from flexible colonoscopy (mean: 64.1 in 2010 and 67.2 in 2023). The volume for colonoscopy remained fairly stable (t = 0.429; p-value = 0.036). This was followed by esophagogastroduodenoscopy (mean: 35.3 in 2010 and 35.5 in 2023), which saw a significant increase in volume (t = 0.890; p-value ≤ 0.001). There was a significant increase in the number of overall upper endoscopic procedures (t = 0.791; p-value ≤ 0.001), while lower endoscopic procedures did not change significantly (t = 0.319; p-value = 0.125). The procedural volume for endoscopic retrograde cholangiography, sigmoidoscopy, cystoscopy/ureteroscopy, laryngoscopy, and bronchoscopy decreased significantly (p-value < 0.05 for all). CONCLUSION: The overall endoscopy volume for general surgery residents has largely remained stable, with a minor increase in esophagogastroduodenoscopy and no change in colonoscopy. Future research should investigate whether simulation-based exercises can bridge the gap between procedural volume and learning curve requirements for endoscopy.

Comparing the influence of intellectual humility, religiosity, and political conservatism on vaccine attitudes in the United States, Canada, and the United Kingdom.

Three studies of US, Canada, and UK respondents examined pro-vaccine attitudes as predicted by intellectual humility, belief in science, religiosity, and political attitudes. Intellectual humility refers to the capacity to understand limits of one's own beliefs and showed strong relationship to pro-vaccine attitudes across samples. Pro-vaccine attitudes were correlated with intellectual humility and negatively correlated with political conservatism and religiosity. Regression models compared overlapping influences of belief predictors on vaccine attitudes. Across countries, intellectual humility was the most consistent predictor of pro-vaccine attitudes when controlling for other beliefs and thinking styles (political conservatism, belief in science, religiosity). In comparison, political conservatism was a significant predictor of vaccine attitudes in regression models on US and Canadian respondents, and religiosity only held as a predictor in regression models in the US sample. We conclude with a discussion of intellectual humility as a predictor of vaccine attitudes and implications for research and persuasion.

The bone marrow is the primary site of thrombopoiesis.

ABSTRACT: Megakaryocytes (MKs) generate thousands of platelets over their lifespan. The roles of platelets in infection and inflammation has guided an interest to the study of extramedullary thrombopoiesis and therefore MKs have been increasingly reported within the spleen and lung. However, the relative abundance of MKs in these organs compared to the bone marrow and the scale of their contribution to the platelet pool in a steady state remain controversial. We investigated the relative abundance of MKs in the adult murine bone marrow, spleen, and lung using whole-mount light-sheet and quantitative histological imaging, flow cytometry, intravital imaging, and an assessment of single-cell RNA sequencing (scRNA-seq) repositories. Flow cytometry revealed significantly higher numbers of hematopoietic stem and progenitor cells and MKs in the murine bone marrow than in spleens or perfused lungs. Two-photon intravital and light-sheet microscopy, as well as quantitative histological imaging, confirmed these findings. Moreover, ex vivo cultured MKs from the bone marrow subjected to static or microfluidic platelet production assays had a higher capacity for proplatelet formation than MKs from other organs. Analysis of previously published murine and human scRNA-seq data sets revealed that only a marginal fraction of MK-like cells can be found within the lung and most likely only marginally contribute to platelet production in the steady state.