Clearing-enabled light sheet microscopy as a novel method for three-dimensional mapping of the sensory innervation of the mouse knee.

A major barrier that hampers our understanding of the precise anatomic distribution of pain sensing nerves in and around the joint is the limited view obtained from traditional two dimensional (D) histological approaches. Therefore, our objective was to develop a workflow that allows examination of the innervation of the intact mouse knee joint in 3D by employing clearing-enabled light sheet microscopy. We first surveyed existing clearing protocols (SUMIC, PEGASOS, and DISCO) to determine their ability to clear the whole mouse knee joint, and discovered that a DISCO protocol provided the most optimal transparency for light sheet microscopy imaging. We then modified the DISCO protocol to enhance binding and penetration of antibodies used for labeling nerves. Using the pan-neuronal PGP9.5 antibody, our protocol allowed 3D visualization of innervation in and around the mouse knee joint. We then implemented the workflow in mice intra-articularly injected with nerve growth factor (NGF) to determine whether changes in the nerve density can be observed. Both 3D and 2D analytical approaches of the light sheet microscopy images demonstrated quantifiable changes in midjoint nerve density following 4 weeks of NGF injection in the medial but not in the lateral joint compartment. We provide, for the first time, a comprehensive workflow that allows detailed and quantifiable examination of mouse knee joint innervation in 3D.

A standardized approach to evaluation and reporting of synovial histopathology in two surgically induced murine models of osteoarthritis.

OBJECTIVE: Synovial pathology has been linked to osteoarthritis (OA) pain in patients. Microscopic grading systems for synovial changes in human OA have been described, but a standardized approach for murine models of OA is needed. We sought to develop a reproducible approach and set of minimum recommendations for reporting of synovial histopathology in mouse models of OA. METHODS: Coronal and sagittal sections from male mouse knee joints subjected to destabilization of medial meniscus (DMM) or partial meniscectomy (PMX) were collected as part of other studies. Stains included Hematoxylin and Eosin (H&E), Toluidine Blue (T-Blue), and Safranin O/Fast Green (Saf-O). Four blinded readers graded pathological features (hyperplasia, cellularity, and fibrosis) at specific anatomic locations. Inter-reader agreement of each feature score was determined. RESULTS: There was acceptable to very good agreement when using 3-4 individual readers. After DMM and PMX, expected medial predominant changes in hyperplasia and cellularity were observed, with fibrosis noted at 12 weeks post-PMX. Synovial changes were consistent from section to section in the mid-joint area. When comparing stains, H&E and T-blue resulted in better agreement compared to Saf-O stain. CONCLUSIONS: To account for the pathologic and anatomic variability in synovial pathology and allow for a more standardized evaluation that can be compared across studies, we recommend evaluating a minimum set of 3 pathological features at standardized anatomic areas. Further, we suggest reporting individual feature scores separately before relying on a single summed "synovitis" score. H&E or T-blue are preferred, inter-reader agreement for each feature should be considered.

Post-transcriptional (re)programming of B lymphocyte development: From bench to bedside?

Hematopoiesis, a process which generates blood and immune cells, changes significantly during mammalian development. Definitive hematopoiesis is marked by the emergence of long-term hematopoietic stem cells (HSCs). Here, we will focus on the post-transcriptional differences between fetal liver (FL) and adult bone marrow (ABM) HSCs. It remains unclear how or why exactly FL HSCs transition to ABM HSCs, but we aim to leverage their differences to revive an old idea: in utero HSC transplantation. Unexpectedly, the expression of certain RNA-binding proteins (RBPs) play an important role in HSC specification, and can be employed to convert or reprogram adult HSCs back to a fetal-like state. Among other features, FL HSCs have a broad differentiation capacity that includes the ability to regenerate both conventional B and T cells, as well as innate-like or unconventional lymphocytes such as B-1a and marginal zone B (MzB) cells. This chapter will focus on RNA binding proteins, namely LIN28B and IGF2BP3, that are expressed during fetal life and how they promote B-1a cell development. Furthermore, this chapter considers a potential clinical application of synthetic co-expression of LIN28B and IGF2BP3 in HSCs.

Retinal Pigment Epithelial Adenoma: Initial Treatment Outcomes following Episcleral Brachytherapy.

Introduction: We aim to explore the safety and efficacy of episcleral brachytherapy as a primary management option for eyes with retinal pigment epithelial (RPE) adenoma. Methods: Retrospective chart review of the demographic, clinical, ancillary, and postoperative outcome data of patients with RPE adenoma in 2 tertiary referral centers. Tumor regression, final visual acuity, and complications were assessed. Results: Five patients (3 females and 2 males) were included. Four of the 5 eyes had peripheral and mid-peripheral lesions, while one tumor was juxtapapillary. Three eyes were treated with ruthenium-106 (100 Gray), and 2 received iodine-125 episcleral plaques (85 Gray). All eyes showed clinical and imaging-based evidence of regression. Four eyes had stable or improved visual acuity, while 1 eye exhibited one line loss of visual acuity due to radiation retinopathy. Local recurrence was not observed in any eye over a median follow-up of 24 (range 6-112) months. Conclusions: Episcleral brachytherapy is an effective management option for select cases of RPE adenoma that is capable of achieving tumor regression while maintaining favorable visual acuity. The initial safety profile of brachytherapy is good without significant vision-compromising complications.

Herpesvirus Entry Mediator as an Immune Checkpoint Target and a Potential Prognostic Biomarker in Myeloid and Lymphoid Leukemia.

Herpesvirus entry mediator (HVEM) is a molecular switch that can modulate immune responses against cancer. The significance of HVEM as an immune checkpoint target and a potential prognostic biomarker in malignancies is still controversial. This study aims to determine whether HVEM is an immune checkpoint target with inhibitory effects on anti-tumor CD4+ T cell responses in vitro and whether HVEM gene expression is dysregulated in patients with acute lymphocytic leukemia (ALL). HVEM gene expression in tumor cell lines and peripheral blood mononuclear cells (PBMCs) from ALL patients and healthy controls was measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Tumor cells were left untreated (control) or were treated with an HVEM blocker before co-culturing with CD4+ T cells in vitro in a carboxyfluorescein succinimidyl ester (CFSE)-dependent proliferation assay. HVEM expression was upregulated in the chronic myelogenous leukemia cell line (K562) (FC = 376.3, p = 0.086) compared with normal embryonic kidney cells (Hek293). CD4+ T cell proliferation was significantly increased in the HVEM blocker-treated K562 cells (p = 0.0033). Significant HVEM differences were detected in ALL PBMCs compared with the controls, and these were associated with newly diagnosed ALL (p = 0.0011) and relapsed/refractory (p = 0.0051) B cell ALL (p = 0.0039) patients. A significant differentiation between malignant ALL and the controls was observed in a receiver operating characteristic (ROC) curve analysis with AUC = 0.78 ± 0.092 (p = 0.014). These results indicate that HVEM is an inhibitory molecule that may serve as a target for immunotherapy and a potential ALL biomarker.

The sensitivities and adaptive capacity of public lands visitors.

Lands and waters administered by governing entities for public use (i.e., "public lands") are subject to changing social and ecological conditions (e.g., overcrowding, drought). Public lands managers are often tasked with addressing these changes while balancing conservation goals and public use mandates, and their decisions can significantly and inequitably impact visitor sensitivities to different types of exposures. To gain insights into visitor sensitivities and their adaptive capacity to mitigate the impacts of exposures, we draw upon a comprehensive monitoring effort conducted in collaboration with the U.S. Fish and Wildlife Service (Service) to understand visitor experiences on national wildlife refuges (refuges). We collected data from 10,556 visitors to 68 refuges during 2018-2019, then segmented respondents into unique visitor types based on their frequency of visiting "this refuge" where they were contacted, their participation across different activities at that refuge, and visits to other public lands for purposes of their primary activity, all during the 12 months prior to being contacted. We then explored differences among the resulting visitor types in their (a) purpose of visit, (b) satisfaction with opportunities during their visit, and (c) demographic characteristics. Finally, we used external data sources to explore the sensitivities and adaptive capacity of visitors' home communities. Our approach identified eight types of visitors with distinct sensitivities and adaptive capacities. For example, the type categorized as "most sensitive" due to activity specialization and site dependency was more likely to engage in activities (e.g., fishing, hunting) that may be subsistence uses of public lands and more often lived in communities with reduced adaptive capacity. Our assessment supports public lands decision-making by helping to understand and address social inequities that may arise or be exacerbated by rapidly changing conditions.

The Impact of Pulmonary Rehabilitation on Mental Health and Quality of Life in Patients With Chronic Obstructive Pulmonary Disease (COPD): A Narrative Review.

Chronic obstructive pulmonary disease (COPD) is a complex, prevalent, debilitating, and degenerative disease that affects a large population, and the treatment options for the patients are limited. Although progress has been made in COPD pathogenesis, etiology, and management, there is still an unmet need to develop novel therapies. COPD management has recently seen a focus on a multidisciplinary pulmonary rehabilitation approach to help patients manage the disease better. This review primarily focuses on the role of pulmonary rehabilitation as a novel therapeutic strategy for treating and managing COPD, which is known to decrease patients' quality of life. Disease management and the beneficial effects of pulmonary rehabilitation in COPD are discussed. Subsequently, different methods that are employed in pulmonary rehabilitation are examined, including oxygen therapy, exercise, meditation, and education, emphasizing how they can help patients better manage COPD. Pathophysiology and the effect of pulmonary rehabilitation on the cellular level, such as the release of perforins and Th1 and Th17 cytokines, are also explored. The link between exercise and meditation during pulmonary rehabilitation therapy, which promotes repairing affected organs, is emphasized. Future perspectives on personalized medicine and its use in conjunction with pulmonary rehabilitation are also outlined. In conclusion, pulmonary rehabilitation holds significant promise for the management of COPD by addressing the present limitations of treatment. However, further research is essential to overcome and optimize treatment strategies for COPD patients.

Dendritic Cell-Based Immunity: Screening of Dendritic Cell Subsets in Breast Cancer-Bearing Mice.

Background: Breast cancer (BC) is the most devastating disease, particularly the lethal invasive form. It is the most underlying cause of death among women worldwide. The expansion of BC is controlled by a variety of alterations in the tumor cells themselves, in addition to the state of the immune system, which has a direct influence on the tumor microenvironment. Numerous receptors expressed by T-cells interact with ligands on antigen-presenting cells to provide activation signals results in mounting effector anti-tumor T-cell responses. On the other hand, there is a dearth of information about the actual interactions and reactions of T-cells and dendritic cells (DCs) all through the progression of tumor development. Aim: Immune system response against BC was investigated through tumor induction in mice. The size and volume of the tumor were calculated. Moreover, the phenotypical profile of T-cells and DCs from lymph nodes (LN) and spleens of BC-bearing mice was investigated. In addition, the levels of Transforming growth factor-ß, Interferon-gamma (IFN-γ), Interleukin IL-2, IL-10, IL-4, IL-12, and tumor necrosis factor (TNF)-α were determined. Materials and Methods: MDA231 cells were utilized to induce BC in 30 white BALB/C mice, whereas the other 30 mice acted as healthy controls and were not treated with any cancer-causing agents. The impact of malignancy was evaluated using flow cytometry based on the marking surface molecules, as well as the titer of specific cytokines of the mice's LN culture using the ELISA method. These cytokines included transforming growth factor-ß (TGF-ß), IFN-γ, IL-2, IL -10, IL -4, IL -12, and TNF-α. Results: The findings showed that the maturation of DCs was inhibited, followed by an accumulation of immature DCs. These immature DCs increase the release of TGF-ß and cytokines like IL-10 and inhibit the release of IFN-γ and IL-12 in the culture supernatant of nodal lymph and spleen suspension of BC-bearing mice compared to control. In addition, there was a low expression of CD80 and CD86 on DCs, which indicates a low maturation process. Conclusion: According to the findings, the tumor microenvironment may have been responsible for preventing the maturation of DCs. This, in turn, weakened the immune response and facilitated the ability of the tumor to proliferate. Furthermore, the tumor microenvironment increased the number of immature DCs by inhibiting their stimulation by overexpression of TGF-ß-produced by regulatory T lymphocytes and stimulation of tumor cells. In addition, the tumor microenvironment stimulated the secretion of cytokines such as IL-10, and CD4 and decreased the secretion of IFN-γ-and IL-12 in tumor-induced mice cultured LN and spleen.

Recommendations For a Standardized Approach to Histopathologic Evaluation of Synovial Membrane in Murine Models of Experimental Osteoarthritis.

Background: Synovial pathology has been linked to osteoarthritis (OA) pain in patients. Microscopic grading systems for synovial changes in human OA have been described, but a standardized approach for murine models of OA is needed. We sought to develop a reproducible approach and set of minimum recommendations for synovial histopathology in mouse models of OA. Methods: Coronal and sagittal sections from male mouse knee joints subjected to destabilization of medial meniscus (DMM) or partial meniscectomy (PMX) were collected as part of other studies. Stains included Hematoxylin and Eosin (H&E), Toluidine Blue (T-Blue) and Safranin O/Fast Green (Saf-O). Four blinded readers graded pathological features (hyperplasia, cellularity, and fibrosis) at specific anatomic locations in the medial and lateral compartments. Inter-reader reliability of each feature was determined. Results: There was acceptable to very good agreement between raters. After DMM, increased hyperplasia and cellularity and a trend towards increased fibrosis were observed 6 weeks after DMM in the medial locations, and persisted up to 16 weeks. In the PMX model, cellularity and hyperplasia were evident in both medial and lateral compartments while fibrotic changes were largely seen on the medial side. Synovial changes were consistent from section to section in the mid-joint area mice. H&E, T-blue, and Saf-O stains resulted in comparable reliability. Conclusions: To allow for a standard evaluation that can be implemented and compared across labs and studies, we recommend using 3 readers to evaluate a minimum set of 3 pathological features at standardized anatomic areas. Pre-defining areas to be scored, and reliability for each pathologic feature should be considered.

Orbital blood vessels changes on color duplex imaging in diabetics with and without diabetic retinopathy.

To compare changes in ophthalmic artery (OA) and its branches in diabetics with and without diabetic retinopathy (DR) using color duplex imaging (CDI), and to correlate these changes with the disease variables. 60 eyes of 60 diabetic patients were enrolled, divided into 3 groups: without DR (Group A), with Non-Proliferative DR (Group B) and with Proliferative DR (PDR) (Group C). Laboratory testing including HbA1c was done. Patients underwent CDI, by which OA, Central Retinal Artery (CRA) and Ciliary Arteries were identified; for each of them we measured Peak systolic velocity (PSV), End Diastolic velocity (EDV) and Resistivity Index (RI). Results were compared to clinical, laboratory and fundus examination. OA EDV was significantly lower and OA RI was found to be significantly higher in Group C (p = 0.027 and 0.025 respectively). CRA PSV and EDV were significantly lower in Group C (p = 0.017 and 0.001 respectively). PCA RI was significantly higher in Group C (p = 0.008). HbA1c was negatively correlated with CRA PSV (p = 0.041), also it was negatively correlated with CRA EDV (p = 0.0001), as well as with PCA EDV (p = 0.002). There was direct significant correlation between HbA1c and PCA RI (p = 0.012). Duration since diagnosis was negatively correlated with CRA EDV (p = 0.004). Multivariate linear regression showed that DR is an independent predictor for low OA EDV, high OA RI, low CRA EDV and high PCA RI. DR is an independent risk factor for orbital and ocular vessels flow alteration, thus can be used as a prognostic tool in diabetic patients. CDI can be reliably used in diabetics to predict early changes or progression of DR.

Sexual dimorphism of the synovial transcriptome underpins greater PTOA disease severity in male mice following joint injury.

OBJECTIVE: Osteoarthritis (OA) is a disease with sex-dependent prevalence and severity in both human and animal models. We sought to elucidate sex differences in synovitis, mechanical sensitization, structural damage, bone remodeling, and the synovial transcriptome in the anterior cruciate ligament rupture (ACLR) mouse model of post-traumatic OA (PTOA). DESIGN: Male and female 12-week-old C57/BL6J mice were randomized to Sham or noninvasive ACLR with harvests at 7d or 28d post-ACLR (n = 9 per sex in each group - Sham, 7d ACLR, 28d ACLR). Knee hyperalgesia, mechanical allodynia, and intra-articular matrix metalloproteinase (MMP) activity (via intravital imaging) were measured longitudinally. Trabecular and subchondral bone (SCB) remodeling and osteophyte formation were assessed by µCT. Histological scoring of PTOA, synovitis, and anti-MMP13 immunostaining were performed. NaV1.8-Cre;tdTomato mice were used to document localization and sprouting of nociceptors. Bulk RNA-seq of synovium in Sham, 7d, and 28d post-ACLR, and contralateral joints (n = 6 per group per sex) assessed injury-induced and sex-dependent gene expression. RESULTS: Male mice exhibited more severe joint damage at 7d and 28d and more severe synovitis at 28d, accompanied by 19% greater MMP activity, 8% lower knee hyperalgesia threshold, and 43% lower hindpaw withdrawal threshold in injured limbs compared to female injured limbs. Females had injury-induced catabolic responses in trabecular and SCB, whereas males exhibited 133% greater normalized osteophyte volume relative to females and sclerotic remodeling of trabecular and SCB. NaV1.8+ nociceptor sprouting in SCB and medial synovium was induced by injury and comparable between sexes. RNA-seq of synovium demonstrated similar injury-induced transcriptomic programs between the sexes at 7d, but only female mice exhibited a transcriptomic signature indicative of synovial inflammatory resolution by 28d, whereas males had persistent pro-inflammatory, pro-fibrotic, pro-neurogenic, and pro-angiogenic gene expression. CONCLUSION: Male mice exhibited more severe overall joint damage and pain behavior after ACLR, which was associated with persistent activation of synovial inflammatory, fibrotic, and neuroangiogenic processes, implicating persistent synovitis in driving sex differences in murine PTOA.

Apoptosis induction in human hepatoma cell line HepG2 cells by trans- Anethole via activation of mitochondria-mediated apoptotic pathways.

trans-Anethole a valuable compound derived from star anise widely used by ethnic tribals to manage numerous human diseases. In this study antiproliferative activities of trans-Anethole towards human liver cancer (HepG2), cervical cancer (HeLa) and breast cancer (MCF-7) cells were explored. trans-Anethole showed free radical scavenging potential as assessed by DNA nicking assay. trans-Anethole exhibited strong antiproliferative potential towards HepG2 cells compared to other cell lines. trans-Anethole strongly induced apoptosis in HepG2 cells by significantly upregulating the protein expressions of p53, Caspase-3 and Caspase-9 were assessed by western blotting analysis which highlighted apoptosis-inducing capacity of trans-Anethole against HepG2 cells. Rt-qPCR analysis revealed that trans- Anethole upregulated p53, caspase - 3 and - 9 in comparison to untreated HepG2 cancer cells. Moreover, trans-Anethole provoked the generation of ROS and disruption of MMP. Our research suggests that trans-Anethole may have a significant anticancer therapeutic potential for treating liver cancer.

Mathematical analysis of radius and length of CNTs on flow of nanofluid over surface with variable viscosity and joule heating.

The transfer of heat is a phenomenon that is significant in a variety of contexts due to the different ways in which it may be utilized in industrial settings. To increase the rate at which heat is transferred, carbon nanotubes (CNTs), which can either be single-wall or multi-walled, are suspended in base fluids, and the resulting mixture is referred to as a "nanofluid. This study looks at how heat transfers through nanofluids that are suspended in carbon nanotubes with different lengths and radii over a stretching surface. It also looks at how changing viscosity and joule heating affect motion. Water is taken as base fluid. This study looks at both carbon nanotubes with one wall and those with more than one. The flow is governed by a series of partial differential equations, which, to control the flow, are transformed into a series of nonlinear ordinary differential equations. Similarity transformation is used to convert the obtained nonlinear ordinary differential equations and accompanying boundary conditions into a form that is dimensionless. To numerically solve the transformed equation, RK-4 with shooting method is used. Graphs and in-depth discussions are used to look at how velocity and temperature profiles are affected by the leading variables. The expression for skin friction and local Nusselt number are written down and graphs show how these two numbers change for different parameter values. The temperature profile goes down when the viscosity parameter goes down, but the velocity profile goes up. When the magnetic parameter goes up, the velocity profile f'(η), goes down, but the velocity profile g(η) and temperature θ(η) both go up at the same time. The rate of heat transfer increases with the addition of φ and S. When the suction parameter (S = 2.1) with 1% of φ is used, it is reported that rate of heat transfer increases by 1.135% for Single walled and 1.275% for Multi Walled carbon nanotubes. To determine whether or not the proposed numerical model is legitimate, a comparison is made between the current results and those that have previously been published.

Analysis of Monkeypox viral infection with human to animal transmission via a fractional and Fractal-fractional operators with power law kernel.

Monkeypox (MPX) is a global public health concern. This infectious disease affects people all over the world, not just those in West and Central Africa. Various approaches have been used to study epidemiology, the source of infection, and patterns of transmission of MPX. In this article, we analyze the dynamics of MPX using a fractional mathematical model with a power law kernel. The human-to-animal transmission is considered in the model formulation. The fractional model is further reformulated via a generalized fractal-fractional differential operator in the Caputo sense. The basic mathematical including the existence and uniqueness of both fractional and fractal-fractional problems are provided using fixed points theorems. A numerical scheme for the proposed model is obtained using an efficient iterative method. Moreover, detailed simulation results are shown for different fractional orders in the first stage. Finally, a number of graphical results of fractal-fractional MPX transmission models are presented showing the combined effect of fractal and fractional orders on model dynamics. The resulting simulations conclude that the new fractal-fractional operator added more biological insight into the dynamics of illness.

Age-Associated Changes in Knee Osteoarthritis, Pain-Related Behaviors, and Dorsal Root Ganglia Immunophenotyping of Male and Female Mice.

OBJECTIVE: Osteoarthritis (OA) is a leading cause of chronic pain, yet OA pain management remains poor. Age is the strongest predictor of OA development, and mechanisms driving OA pain are unclear. We undertook this study to characterize age-associated changes in knee OA, pain-related behaviors, and dorsal root ganglion (DRG) molecular phenotypes in mice of both sexes. METHODS: Male or female C57BL/6 mice 6 or 20 months of age were evaluated for histopathologic knee OA, pain-related behaviors, and L3-L5 DRG immune characterization via flow cytometry. DRG gene expression in older mice and humans was also examined. RESULTS: Male mice at 20 months of age had worse cartilage degeneration than 6-month-old mice. Older female mouse knees showed increased cartilage degeneration but to a lesser degree than those of male mice. Older mice of both sexes had worse mechanical allodynia, knee hyperalgesia, and grip strength compared to younger mice. For both sexes, DRGs from older mice showed decreased CD45+ cells and a significant increase in F4/80+ macrophages and CD11c+ dendritic cells. Older male mouse DRGs showed increased expression of Ccl2 and Ccl5, and older female mouse DRGs showed increased Cxcr4 and Ccl3 expression compared to 6-month-old mouse DRGs, among other differentially expressed genes. Human DRG analysis from 6 individuals >80 years of age revealed elevated CCL2 in men compared to women, whereas CCL3 was higher in DRGs from women. CONCLUSION: We found that aging in male and female mice is accompanied by mild knee OA, mechanical sensitization, and changes to immune cell populations in the DRG, suggesting novel avenues for development of OA therapies.

Piezo2 expressing nociceptors mediate mechanical sensitization in experimental osteoarthritis.

Non-opioid targets are needed for addressing osteoarthritis pain, which is mechanical in nature and associated with daily activities such as walking and climbing stairs. Piezo2 has been implicated in the development of mechanical pain, but the mechanisms by which this occurs remain poorly understood, including the role of nociceptors. Here we show that nociceptor-specific Piezo2 conditional knock-out mice were protected from mechanical sensitization associated with inflammatory joint pain in female mice, joint pain associated with osteoarthritis in male mice, as well as both knee swelling and joint pain associated with repeated intra-articular injection of nerve growth factor in male mice. Single cell RNA sequencing of mouse lumbar dorsal root ganglia and in situ hybridization of mouse and human lumbar dorsal root ganglia revealed that a subset of nociceptors co-express Piezo2 and Ntrk1 (the gene that encodes the nerve growth factor receptor TrkA). These results suggest that nerve growth factor-mediated sensitization of joint nociceptors, which is critical for osteoarthritic pain, is also dependent on Piezo2, and targeting Piezo2 may represent a therapeutic option for osteoarthritis pain control.

Growth, physiological, and molecular responses of three phaeophyte extracts on salt-stressed pea (Pisum sativum L.) seedlings.

BACKGROUND: Seaweeds are a viable bioresource for suffering plants against salt stress, as they abundant in nutrients, hormones, vitamins, secondary metabolites, and many other phytochemicals that sustain plants' growth under both typical and stressful situations. The alleviating capacity of extracts from three brown algae (Sargassum vulgare, Colpomenia sinuosa, and Pandia pavonica) in pea (Pisum sativum L.) was investigated in this study. METHODS: Pea seeds were primed for 2 h either with seaweed extracts (SWEs) or distilled water. Seeds were then subjected to salinity levels of 0.0, 50, 100, and 150 mM NaCl. On the 21st day, seedlings were harvested for growth, physiological and molecular investigations. RESULTS: SWEs helped reduce the adverse effects of salinity on pea, with S. vulgare extract being the most effective. Furthermore, SWEs diminished the effect of NaCl-salinity on germination, growth rate, and pigment content and raised the osmolytes proline and glycine betaine levels. On the molecular level, two low-molecular-weight proteins were newly synthesized by the NaCl treatments and three by priming pea seeds with SWEs. The number of inter-simple sequence repeats (ISSR) markers increased from 20 in the control to 36 in 150 mM NaCl-treated seedlings, including four unique markers. Priming with SWEs triggered more markers than the control, however about ten of the salinity-induced markers were not detected following seed priming before NaCl treatments. By priming with SWEs, seven unique markers were elicited. CONCLUSION: All in all, priming with SWEs alleviated salinity stress on pea seedlings. Salinity-responsive proteins and ISSR markers are produced in response to salt stress and priming with SWEs.

Factors Influencing the Length of Hospital Stay Among Pediatric COVID-19 Patients at Queen Rania Al Abdullah Hospital for Children: A Cross-Sectional Study.

Background COVID-19 caused by SARS-CoV-2 is a worldwide epidemic. Children are less commonly infected and have less severe symptoms than adults. However, they are at risk for COVID-19-associated severe sickness and hospitalization. The duration of stay is a major driver of effective health treatment during hospitalization; thus, it is only logical to attempt to comprehend the factors influencing the length of stay (LOS) for these patients, particularly in light of the ongoing pandemic caused by the new SARS-CoV-2 virus. As predictors of hospital LOS, several variables, including age, gender, disease severity, hospital mortality, insurance type, and hospital location, have been discovered. In our study, we focused on the severity of the patient's condition, the presence of comorbidities, and the necessary therapeutic regimen to predict the duration of stay. This study aimed to answer the following questions: If a patient has comorbidity and has COVID-19 requiring hospital treatment, will the patient's comorbidity elongate the duration of stay at the hospital for further management in the pediatric age group? What are the risk factors that play a significant role in the hospital stay duration in pediatrics? Methodology We gathered data from 100 hospitalized children aged up to 14 years who tested positive for COVID-19, which was not specific to variants of SARS-CoV-2, over 24 months (February 2020-February 2022) at Queen Rania Al Abdullah Hospital for Children, one of the Health Care Accreditation Council accredited facilities. Clinical symptoms, signs, oxygen demand, imaging study results, laboratory data, and usage of corticosteroid and antiviral medication were all taken from patients' medical records. There were no limitations in taking the sample of patients. All patients in the duration mentioned were included. Results Clinical data of 100 COVID-19-positive pediatric patients were analyzed; 52% of the patients had associated chronic illnesses, while 48% were medically free. The longest duration of LOS was 28 days, the shortest was one day, the median was eight days, and five days was the most frequent among patients owing to 21% of patients, using mean descriptive statistics. We compared LOS to having or not having comorbidities. The mean LOS of patients with the comorbid disease was 6.15 days, with a maximum of 28 days, while for patients without chronic illnesses, the mean was 4.81 days with a maximum of 14 days. The significance was 0.07. Our results also showed a significant correlation between using steroids and LOS, as it had an advantageous effect by decreasing it with a significance value of 0.04. Having abnormal findings on chest computed tomography (CT) scan was also associated with increased LOS with a significant value of 0.00. Conclusions According to our research, there was no direct association between comorbidity and hospital LOS, which is counterintuitive, as it was influenced by multiplayers of variables such as using steroids, which decreased the LOS, and abnormal findings on chest CT, which resulted in lengthening of the hospital stay. Our findings cannot be proven without further research and a larger patient sample.

Golgi signalling proteins GOLPH3, MYO18A, PITPNC1 and RAB1B: implications in prognosis and survival outcomes of AML patients.

BACKGROUND: The role of different Golgi signalling proteins remains unexplored in the progression and spread of acute myeloid leukaemia (AML), whom all interact together in a way that facilitates proliferation and differentiation of myeloid lineage cells. OBJECTIVE: Since Golgi apparatus acts as master brain in membrane trafficking and signalling events that affect cell polarity necessary for migration, division, or differentiation; this study aims to explore the association between signalling proteins and the diagnosis, prognosis, and survival of AML patients. MATERIAL AND METHODS: This study comprised 70 newly diagnosed AML patients and 20 healthy controls to investigate the serum levels of signalling proteins; Golgi Phosphoprotein 3 (GOLPH3), Myosin 18A (MYO18A), Cytoplasmic Phosphatidylinositol Transfer Protein 1 (PITPNC1) and Ras-Associated Binding Protein 1B (RAB1B). RESULTS: AML patients showed higher serum levels of GOLPH3, MYO18A, PITPNC1 and RAB1B when compared to control (p < 0.001). A significant negative correlation was found between the patients' overall survival and GOLPH3 (p = 0.001), MYO18A (p = 0.011), PITPNC1 (p = 0.001) and RAB1B (p = 0.042). Results were confirmed by Kaplen-Meier survival analysis showing lower survival estimates in patients with higher GOLPH3 (p = 0.014), MYO18A (p = 0.047), PITPNC1 (p = 0.008) and RAB1B (p = 0.033) serum levels. CONCLUSION: GOLPH3, MYO18A, PITPNC1 and RAB1B maybe promising diagnostic and prognostic biomarkers in AML patients.