Cell Aggregation Research Articles

Histopathological Analysis of Lipopolysaccharide-Induced Liver Inflammation and Thrombus Formation in Mice: The Protective Effects of Aspirin

Hepatitis, a significant medical concern owing to its potential to cause acute and chronic liver disease, necessitates early intervention. In this study, we aimed to elucidate the histopathological features of lipopolysaccharide-induced hepatitis in mice, focusing on tissue alterations. The results demonstrated that hepatocytes exhibited decreased eosin staining, indicating cellular shrinkage, whereas sinusoids were swollen with blood cells. Detailed electron microscope analysis identified these blood cells as leukocytes and erythrocytes, which confirmed a thrombus formation within the liver. Pre-treatment with aspirin significantly attenuated these pathological changes, including reductions in inflammatory markers such as C-reactive protein, interleukin-1β, and tumor necrosis factor-alpha. These findings highlight aspirin’s anti-inflammatory and antiplatelet effects in mitigating liver inflammation and thrombus formation. In this study, we highlighted the potential of aspirin as a therapeutic agent for liver inflammation, in addition to providing insights into hepatocyte alterations and sinusoidal blood cell aggregation in liver inflammation. Aspirin, through the protection of endothelial cells and reduction of cytokine levels, may have broader applications in managing liver disease and other systemic inflammatory conditions. This emphasizes its value in prevention and therapy.

Histopathological Insights into Granulomatous Lesions: A Comprehensive Study

ABSTRACT Background: Granulomatous inflammation is a persistent inflammatory reaction marked by the presence of aggregations of active histiocytes, epithelioid cells, and multinucleate giant cells. This phenomenon is reported in several viral and non-infectious diseases and is classified as a type IV delayed hypersensitivity response. Research Materials and Procedures: The present cross-sectional study was conducted in the histopathology section of the Department of Pathology, MMIMSR, Mullana, from October 2022 to October 2023. A total of 100 cases presenting with granulomatous lesions were analyzed. Detailed clinical data, including age, sex, site of the lesion, and duration of disease, were recorded. Histopathological examination was performed using Hematoxylin and Eosin (H&E) stain, with additional special stains like Ziehl-Neelsen and Periodic Acid-Schiff (PAS) applied where necessary. Materials and Methods: This investigation was conducted at the histology division of the Department of Pathology, MMIMSR, Mullana, between October 2022 and October 2023. The analysis included a total of 100 patients that had granulomatous lesions. Precise clinical information, such as age, gender, location of the lesion, and length of illness, was documented. The histological analysis was conducted using hematoxylin and eosin (H and E) stain, along with supplementary specific stains, such as Ziehl-Neelsen and periodic acid-Schiff (PAS) when needed. Results: Lesions of granulomatous nature were more common in individuals in their third and fourth decades of life, with a little higher occurrence in females (male: female ratio = 1:1.43). The primary location of infection was the gastrointestinal system, accounting for 16% of cases, followed by the urogenital tract at 15% and soft tissues at 13%. Granulomatous pathology-not otherwise specified (GP-NOS) was the most prevalent histological category, representing 52% of cases. It was followed by granulomatous pathology-tubercular (GP-TB) at 15%, granulomatous pathology-leprosy (GP-L) at 3%, and one case of granulomatous pathology-fungal (GP-F). The etiology of 25% of cases was attributed to tuberculosis, whereas in 3% of instances, it was diagnosed as leprosy. The main kind of neoplasms seen were epithelioid cell granulomas, with caseous necrosis being the most prevalent form of necrosis. About 10% of patients had fibrosis, and Langhans giant cells were often documented. Conclusion: Histopathology, with the use of certain stains, continues to be the most reliable method for identifying and classifying granulomatous lesions. Despite the broad use of Bacillus Calmette-Guerin (BCG) vaccine, infections, including tuberculosis (TB), remain the primary cause of granulomatous lesions in our community.

TNBS colitis induces architectural changes and alpha-synuclein overexpression in mouse distal colon: A morphological study.

Alpha-synuclein (α-syn) is widely expressed in presynaptic neuron terminals, and its structural alterations play an important role in the pathogenesis of Parkinson's disease (PD). Aggregated α-syn has been found in brain, in the peripheral nerves of the enteric nervous system (ENS) and in the intestinal neuroendocrine cells during synucleinopathies and inflammatory bowel disorders. In the present study, we evaluated the histomorphological features of murine colon with 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis, a common model of colitis. Thereafter, we investigated the expression of α-syn, Toll-like receptor 4 (TLR4), choline acetyltransferase (ChAT), vasoactive intestinal peptide (VIP), tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP), and calcitonin-like receptor (CALCR). Finally, we investigated the presence of phosphorylated α-syn (pS129 α-syn) aggregates and their relationship with inflammatory cells. Colon from TNBS mice showed an increase in inflammatory cells infiltrate and significative changes in the architecture of the intestinal mucosa. α-Syn expression was significantly higher in inflamed colon. VIP was increased in both the mucosa and muscularis externa of TNBS mice, while TH, CGRP, and CALCR were significantly reduced in TNBS mice. Amyloid aggregates of pS129 α-syn were detectable in the ENS, as in the macrophages around the glands of the mucosa correlating with the markers of inflammation. This study describes - for the first time - the altered expression of α-syn and the occurrence of amyloid α-syn aggregates in the inflammatory cells under colitis, supporting the critical role of bowel inflammation in synucleinopathies and the involvement of α-syn in IBD.

Understanding the development of tuberculous granulomas: insights into host protection and pathogenesis, a review in humans and animals

Granulomas, organized aggregates of immune cells which form in response to Mycobacterium tuberculosis (Mtb), are characteristic but not exclusive of tuberculosis (TB). Despite existing investigations on TB granulomas, the determinants that differentiate host-protective granulomas from granulomas that contribute to TB pathogenesis are often disputed. Thus, the goal of this narrative review is to help clarify the existing literature on such determinants. We adopt the a priori view that TB granulomas are host-protective organelles and discuss the molecular and cellular determinants that induce protective granulomas and those that promote their failure. While reports about protective TB granulomas and their failure may initially seem contradictory, it is increasingly recognized that either deficiencies or excesses of the molecular and cellular components in TB granuloma formation may be detrimental to the host. More specifically, insufficient or excessive expression/representation of the following components have been reported to skew granulomas toward the less protective phenotype: (i) epithelioid macrophages; (ii) type 1 adaptive immune response; (iii) type 2 adaptive immune response; (iv) tumor necrosis factor; (v) interleukin-12; (vi) interleukin-17; (vii) matrix metalloproteinases; (viii) hypoxia in the TB granulomas; (ix) hypoxia inducible factor-1 alpha; (x) aerobic glycolysis; (xi) indoleamine 2,3-dioxygenase activity; (xii) heme oxygenase-1 activity; (xiii) immune checkpoint; (xiv) leukotriene A4 hydrolase activity; (xv) nuclear-factor-kappa B; and (xvi) transforming growth factor-beta. Rather, more precise and timely coordinated immune responses appear essential for eradication or containment of Mtb infection. Since there are several animal models of infection with Mtb, other species within the Mtb complex, and the surrogate Mycobacterium marinum – whether natural (cattle, elephants) or experimental (zebrafish, mouse, guinea pig, rabbit, mini pig, goat, non-human primate) infections – we also compared the TB granulomatous response and other pathologic lung lesions in various animals infected with one of these mycobacteria with that of human pulmonary TB. Identifying components that dictate the formation of host-protective granulomas and the circumstances that result in their failure can enhance our understanding of the macrocosm of human TB and facilitate the development of novel remedies – whether they be direct therapeutics or indirect interventions – to efficiently eliminate Mtb infection and prevent its pathologic sequelae.

In vitro culture of leukemic cells in collagen scaffolds and carboxymethyl cellulose-polyethylene glycol gel.

Chronic lymphocytic leukemia (CLL) is a common adult leukemia characterized by the accumulation of neoplastic mature B cells in blood, bone marrow, lymph nodes, and spleen. The disease biology remains unresolved in many aspects, including the processes underlying the disease progression and relapses. However, studying CLL in vitro poses a considerable challenge due to its complexity and dependency on the microenvironment. Several approaches are utilized to overcome this issue, such as co-culture of CLL cells with other cell types, supplementing culture media with growth factors, or setting up a three-dimensional (3D) culture. Previous studies have shown that 3D cultures, compared to conventional ones, can lead to enhanced cell survival and altered gene expression. 3D cultures can also give valuable information while testing treatment response in vitro since they mimic the cell spatial organization more accurately than conventional culture. In our study, we investigated the behavior of CLL cells in two types of material: (i) solid porous collagen scaffolds and (ii) gel composed of carboxymethyl cellulose and polyethylene glycol (CMC-PEG). We studied CLL cells' distribution, morphology, and viability in these materials by a transmitted-light and confocal microscopy. We also measured the metabolic activity of cultured cells. Additionally, the expression levels of MYC, VCAM1, MCL1, CXCR4, and CCL4 genes in CLL cells were studied by qPCR to observe whether our novel culture approaches lead to increased adhesion, lower apoptotic rates, or activation of cell signaling in relation to the enhanced contact with co-cultured cells. Both materials were biocompatible, translucent, and permeable, as assessed by metabolic assays, cell staining, and microscopy. While collagen scaffolds featured easy manipulation, washability, transferability, and biodegradability, CMC-PEG was advantageous for its easy preparation process and low variability in the number of accommodated cells. Both materials promoted cell-to-cell and cell-to-matrix interactions due to the scaffold structure and generation of cell aggregates. The metabolic activity of CLL cells cultured in CMC-PEG gel was similar to or higher than in conventional culture. Compared to the conventional culture, there was (i) a lower expression of VCAM1 in both materials, (ii) a higher expression of CCL4 in collagen scaffolds, and (iii) a lower expression of CXCR4 and MCL1 (transcript variant 2) in collagen scaffolds, while it was higher in a CMC-PEG gel. Hence, culture in the material can suppress the expression of a pro-apoptotic gene (MCL1 in collagen scaffolds) or replicate certain gene expression patterns attributed to CLL cells in lymphoid organs (low CXCR4, high CCL4 in collagen scaffolds) or blood (high CXCR4 in CMC-PEG).

Analysis of a model describing bacterial colony expansion in radial geometry driven by chemotaxis

Abstract We investigate a recent model proposed in the literature elucidating patterns driven by chemotaxis, similar to viscous fingering phenomena. Notably, this model incorporates a singular advection term arising from a modified formulation of Darcy’s law. It is noteworthy that this type of advection can also be well interpreted as a description of a radial fluid flow source surrounding an aggregation of cells. For the two-dimensional scenario, we establish a precise threshold delineating between blow-up and global solution existence. This threshold is contingent upon the pressure magnitude and the initial total mass of the aggregating cells.

Non-invasive and continuous monitoring of 3D stem cell culture in a bioreactor

Abstract The development and discovery of new drugs and therapy products in regenerative medicine are carried out through multiple stages which involve several strict quality protocols. The use of cell-based model systems is the most relevant alternative to deviate from animal testing. However, the underlying biomanufacturing process in a 3D environment can be prone to failures or unexpected behaviors such as cell growth anomalies or inconsistencies in cell aggregation, which can seriously affect the quality of the result. Therefore, it is necessary to establish a non-invasive and continuous monitoring mechanism that minimizes contamination risks, allows taking early decisions and enables large-scale production in the long term. The results of our embedded machine learning approach show that it is feasible to train machine learning models that can operate on a resourcelimited embedded board with acceptable prediction accuracy for the estimation of the average size of growing stem cell spheroids. Further experiments are needed to investigate the full information potential of the recorded process data.

TssL2 of T6SS2 is required for mobility, biofilm formation, wrinkly phenotype formation, and virulence of Vibrio parahaemolyticus SH112

Type VI secretion system 2 (T6SS2) of Vibrio parahaemolyticus is required for cell adhesion and autophagy in macrophages; however, other phenotypes conferred by this T6SS have not been thoroughly investigated. We deleted TssL2, a key component of T6SS2 assembly, to explore the role of the T6SS2 in environmental adaptation and virulence. TssL2 deletion reduced Hcp2 secretion, suggesting that TssL2 played an important role in activity of functional T6SS2. We found that TssL2 was necessary for cell aggregation, wrinkly phenotype formation, and participates in motility and biofilm formation by regulating related genes, suggesting that TssL2 was essential for V. parahaemolyticus to adapt changing environments. In addition, this study demonstrated TssL2 significantly affected adhesion, cytotoxicity, bacterial colonization ability, and mortality in mice, even the levels of the proinflammatory cytokines IL-6 and IL-8, suggesting that TssL2 was involved in bacterial virulence and immunity. Proteome analysis revealed that TssL2 significantly affected the expression of 163 proteins related to ABC transporter systems, flagellar assembly, biofilm formation, and multiple microbial metabolism pathways, some of which supported the effect of TssL2 on the different phenotypes of V. parahaemolyticus. Among them, the decreased expression of the T3SS1 and T2SS proteins was confirmed by the results of gene transcription, which may be the main reason for the decrease in cytotoxicity. Altogether, these findings further our understanding of T6SS2 components on environmental adaption and virulence during bacterial infection.Key points• The role of T6SS2 in V. parahaemolyticus was far from clear.• TssL2 participates in cell aggregation, wrinkly phenotype formation, motility, and biofilm formation.• TssL2 is essential for cell bacterial colonization, cytotoxicity, virulence, and proinflammatory cytokine production.

The role of the proteosurfaceome and exoproteome in bacterial coaggregation.

Bacterial coaggregation is a critical process in multispecies biofilm formation, driven by specific molecular interactions that facilitate the adhesion and aggregation of bacterial cells. These interactions are essential for the development and persistence of complex microbial communities. This review provides a comprehensive analysis of the roles of the proteosurfaceome and exoproteome in bacterial coaggregation. The proteosurfaceome, comprising surface-bound molecules such as adhesins, drives species-specific interactions crucial for partner recognition and adhesion. In parallel, the exoproteome, particularly extracellular polymeric substances (EPS), enhances aggregate stability by reinforcing structural integrity and facilitating intercellular communication, although its direct role in coaggregation remains to be fully clarified. By integrating these perspectives, this review aims to elucidate how the proteosurfaceome and exoproteome influence bacterial coaggregation, offering insights into their combined impact on microbial community structure and function. Furthermore, we highlight existing knowledge gaps and propose directions for future research.

Enhancing intrinsic TGF-β signaling via heparan sulfate glycosaminoglycan regulation to promote mesenchymal stem cell capabilities and chondrogenesis for cartilage repair

Osteoarthritis burdens patients due to the limited regenerative capacity of chondrocytes. Traditional cartilage repair often falls short, necessitating innovative approaches. Mesenchymal stem cells (MSCs) show promise for regeneration. Heparan sulfate glycosaminoglycans (HS-GAGs) regulate cellular functions, making them a target for cartilage repair. This study highlights how Heparinase III (HepIII) cleaves intact HS-GAGs in bone marrow-derived MSCs (BM-MSCs), enhancing their capabilities and specifically promoting chondrogenesis. HepIII-treated BM-MSCs cultured in a hanging drop device for three days, significantly increased cell number and aggregation into a cell sphere with early chondrogenesis. HepIII promoted BM-MSCs toward chondrogenesis, increasing type II collagen, intact HS-GAGs, and sulfated GAG content, while upregulating chondrogenic and heparan sulfate proteoglycan genes. Treatment with the TGF-β inhibitor (SB-431542) in HepIII-treated BM-MSCs demonstrated enhanced intrinsic transforming growth factor-β (TGF-β) signaling and fibronectin expression. This approach also boosted BM-MSC self-renewal, immunosuppressive potential, and modified acetylated histone signatures, offering a cost-effective strategy for cartilage repair by addressing inflammation, metabolic changes, and the high costs of traditional TGF-β methods. From the results, HepIII-treated BM-MSCs show potential for use in combination with other biopolymers as injectable gels to improve cartilage repair in osteoarthritis patients in the near future.

Ultrafast Self‐Gelling, Adhesive, Anti‐Bacterial Coacervate‐Based Powders for Enhanced Hemostasis and Wound Healing

AbstractUncontrolled hemorrhage, especially in non‐compressible and deep wounds, remains a critical issue in emergency and surgical care. Existing hemostatic powders often lack rapid gelation, mechanical robustness, and adequate adherence, increasing the risk of rebleeding under high‐pressure blood flow. To address these limitations, PQPP, a novel self‐gelling hemostatic material composed of polyacrylamide/quaternized chitosan coacervates and polydopamine nanoparticles is developed. PQPP can rapidly absorb blood within 2 s, undergoes in situ gelation, and forms a robust adhesive hydrogel to effectively seal wounds. Its efficacy stems from the electrostatic adsorption and catechol functional groups of polydopamine nanoparticles. Importantly, PQPP exhibits high burst pressure resistance, excellent blood cell aggregation capability, outstanding biocompatibility, and antibacterial properties. Comprehensive in vitro and in vivo studies, including cytotoxicity and blood compatibility tests, as well as trials in mouse liver, heart, and vascular injury models, demonstrate PQPP's superior hemostatic performance under high‐pressure conditions without causing inflammation. With its rapid gelation, robust adhesion, and mechanical integrity, PQPP represents a promising hemostatic material for immediate wound management in surgical and emergency applications.

The global regulator SpoVG is involved in biofilm formation and stress response in foodborne Staphylococcus aureus

Staphylococcus aureus (S. aureus) is a primary culprit of food poisoning. As a highly adaptable pathogen, S. aureus demonstrates formidable biofilm-forming and stress tolerance capabilities, inducing significant challenges to eradicate food contamination caused by this organism. SpoVG, a regulatory protein in S. aureus, controls the expression of numerous genes. However, its role in biofilm formation and stress response in foodborne S. aureus remains to be elucidated. In this study, we investigated the functions of SpoVG involved in food-related stress responses and biofilm formation in S. aureus RMSA50. The results demonstrated that SpoVG deletion enhanced biofilm formation and resistance to heat and desiccation, while decreased tolerance to oxidative stress. Further analysis revealed that cell aggregation and the accumulation of extracellular DNA (eDNA) may contribute to the enhanced biofilm formation. Real-time quantitative reverse transcription-PCR (RT-qPCR) revealed that the expression levels of nuc and sasC, which are related to cell aggregation and eDNA concentration, were significantly altered in the spoVG mutant. Electrophoretic mobility shift assays (EMSA) confirmed that SpoVG directly binds to the promoter region of nuc and sasC to regulate their expression. These findings suggest that SpoVG may serve as a target to decrease biofilm formation and control S. aureus contamination in the food industry.

Th1 cells are critical tissue organizers of myeloid-rich perivascular activation niches.

Perivascular niche (PVN): myeloid hubs in inflamed mouse and healthy human skinTh1 cells enter, get activated, and leave the PVN within first 24h of tissue entryAntigen-specific signals in the PVN promote the tissue organizing functions of Th1sTh1 cells assemble the PVN via CXCR2-dependent myeloid cell aggregation.

Testing the coordination hypothesis: incompatibilities in aggregative development of an experimentally evolved social amoeba

Abstract Multicellular organisms that form by aggregation of cells arguably do not achieve high levels of complexity. Conflict among the cells is a widely accepted explanation for this, but an alternative hypothesis is that mixing cells of different genotypes leads to failures of coordination, which we call the “coordination hypothesis.” We empirically tested the coordination hypothesis in the social amoeba Dictyostelium discoideum. We mixed D. discoideum clones that had evolved in isolation for generations and acquired mutations that have not been tested against each other by selection. To quantify the effect of incompatibilities, we measured performance in terms of the developmental traits of slug migration and spore production. Importantly, we mixed lines evolved from the same ancestor under conditions that would not select for the evolution of de novo kin recognition. Our results show no evidence of incompatibilities in four traits related to the coordinated movement of slugs toward light in the social amoeba. Spore production was higher than expected in mixtures, in apparent contradiction to the coordination hypothesis. However, we found support for coordination incompatibilities in an interaction between migration and spore production: in mixtures, fewer cells succeeded at both migrating and becoming spores.

Red Blood Cell Morphology Is Associated with Altered Hemorheological Properties and Fatigue in Patients with Long COVID.

SARS-CoV-2 infection adversely affects rheological parameters, particularly red blood cell (RBC) aggregation and deformability, but whether these changes persist in patients suffering from Long-COVID (LC) and whether these changes are related to RBC morphology remain unknown. Venous blood was collected from n = 30 diagnosed LC patients and n = 30 non-LC controls and RBC deformability, RBC aggregation, and hematological parameters were measured. In addition, RBCs were examined microscopically for morphological abnormalities. The mechanical sensitivity index (MS) was assessed in n = 15 LC and n = 15 non-LC samples. Hematological parameters did not differ between the groups. However, LC showed higher aggregation-related parameters. Although RBC deformability was higher in LC, MS, reflecting the functional capacity to deform, was limited in this group. RBCs from LC showed significantly more morphological abnormalities. The extent of morphological abnormalities correlated with MS and the FACIT-Fatigue score of the LC patients. RBCs from LC show a high degree of morphological abnormalities, which might limit the blood flow determining RBC properties and also be related to fatigue symptomatology in LC. Approaches are now needed to understand the underlying cause of these alterations and to ameliorate these permanent changes.

Extended Spherical Diffusion Theory: Electrochemiluminescence Imaging Analysis of Diffusive Molecules from Spherical Biosamples.

Spherical biosamples such as immunobeads, cells, and cell aggregates have been widely used in bioapplications. The bioactivity of individual spherical biosamples in highly sensitive assays and individual analyses must be evaluated in a high-throughput manner. Electrochemiluminescence (ECL) imaging was recently proposed for the high-throughput analysis of diffusive molecules from spherical biosamples. ECL imaging involves the placing of spherical biosamples on a flat electrode filled with a solution. The biosamples produce (or consume) biological/chemical molecules such as H2O2 and O2, which diffuse to form a concentration gradient at the electrode. The ECL signals from the molecules are then measured to obtain the concentration profile, which allows the flux to be estimated, from which their bioactivities can be successfully calculated. However, no studies on theoretical approaches for spherical biosamples on flat surfaces have been conducted using ECL imaging. Therefore, this paper presents a novel spherical diffusion theory for spherical biosamples on a flat surface, which is based on the common spherical diffusion theory and was designated as the extended spherical diffusion theory. First, the concepts behind this theory are discussed. The theory is then validated by comparison with a simulated analysis. The resulting equation successfully expresses the concentration profile for the entire area. The glucose oxidase activity in the hydrogel beads is subsequently visualized using ECL imaging, and the enzymatic product flux is calculated using the proof-of-concept theory. Finally, a time-dependent simulation is conducted to fill the gap between the theoretical and experimental data. This paper presents novel guidelines for this analysis.

The pathogenic APP N-terminal Val225Ala mutation alters tau protein liquid-liquid phase separation and exacerbates synaptic damage.

Amyloid precursor protein (APP) is predominantly located in synapses of neurons and its mutations have been well recognized as the most important genetic causal factor for the familial Alzheimer's disease (AD). While most disease-causal mutations of APP occur within the Aβ-coding region or immediately proximal, the pathological impacts of mutations in the N-terminus of APP protein, which remote from the Aβ sequence, on neuron and synapse are still largely unknown. It was recently reported a pathogenic APP N-terminal Val225Ala mutation (APPV225A) with clinically featuring progressive dementia and typical AD pathologies in brain. In our present study, we further found that APPV225A mutation alters the N-terminal structure of APP, which enhances its binding affinity to tau protein and significantly increases APP-mediated endocytosis. Consequently, APPV225A promotes the uptake of extracellular tau into SH-SY5Y cells, further linking the structural change in APP to intracellular tau accumulation. In addition, APPV225A also notably alters the liquid-liquid phase separation (LLPS)of intracellular tau and intensified tauphosphorylation and aggregation in SH-SY5Y cells. Moreover, APPV225A promote AD-like tau pathology and synaptic damages in human induced pluripotent stem cells (hiPSCs)-derived neural progenitor cells and neurons, as well as in hiPSCs-derived human brain organoids and mouse brain, which can be ameliorated by tau knockdown. Proximity labeling identified several key APPV225A-interacting proteins, including HS3ST3A1, which was shown to directly regulate tau LLPS and phosphorylation. These findings nicely build on our previous work on roles for APP in tau-related pathological phenotypes and further highlight the involvement of N-terminal APP as the key region for both amyloidopathy and tauopathy, two aspects of AD pathogenesis and progression. Our study may also provide a theoretical breakthrough for AD therapy and highlight the important hub roles of APP and making previously neglected N-terminal APP as a potential target for the discovery of novel disease-modifying therapeutic agents against AD, holding significant scientific values and clinical promise.

Spherical Shell Bioprinting to Produce Uniform Spheroids with Controlled Sizes.

Conventional cell spheroid production methods are largely manual, leading to variations in size and shape that compromise consistency and reliability for use in cell-based therapeutic applications. To enhance spheroid production, a spherical shell bioprinting system was implemented, enabling the high-throughput generation of uniform cell spheroids with precisely controlled sizes. The system encapsulates cells within thin alginate hydrogel shells formed through bioprinting and ion crosslinking reactions. Alginate-calcium ion crosslinking created alginate shells that contained gelatin-based bioinks with embedded cells, facilitating spontaneous cell aggregation within the shells and eliminating the need for plastic wells. By adjusting cell concentrations in the alginate-gelatin bioink, we achieved precise control over spheroid size, maintaining a sphericity above 0.94 and size deviations within ±10 µm. This method has been successfully applied to various cell types including cancer cells, fibroblasts, chondrocytes, and epithelial cells, demonstrating its versatility. This scalable approach enhances the reliability of cell therapy and drug screening, offering a robust platform for future biomedical applications.

Isolation and phylogenetic analysis of camel contagious ecthyma virus in Morocco

Camel contagious ecthyma is a highly infectious viral skin disease that affects camels and causes economic losses. This study reports the isolation and the phylogenetic analysis of contagious ecthyma virus among camels in Morocco. The disease was detected in four among fifteen camels with severe papules on the lips and nares. Samples of skin crusts were collected and pooled for virus isolation and titration, PCR testing, and histopathological examination. PCR was used to amplify the B2L gene and the resulting product was sequenced and analyzed genetically. The study's findings indicate the presence of characteristic microscopic changes of poxvirus infection in the examined tissues and the virus was isolated on two cells (lambs testis and Vero) and showed distinct growth patterns. The virus grew rapidly on TA cells and delayed growth on Vero cells.The third passage showed cytopathic effects characterized by cell aggregation. Sequence analysis of the B2L gene revealed 100 % similarity to the camel contagious Ecthyma virus isolated from Ethiopia. The camel virus isolates can be classified into two genetic clades according to the B2L gene sequence: the Asian lineage, which includes isolates from Saudi Arabia, Bahrain, and India, and the African lineage, which includes isolates from the Sudan.In conclusion, this is the first instance of the camel contagious ecthyma virus being identified in North Africa in a herd of camels following exposure to stress. Moreover, the progression of the disease was closely monitored from onset to recovery in a setting without the bacterial complication often observed in the field. The virus exhibited opportunistic behavior, exploiting the stress response. Further studies are warranted to evaluate the pathogenicity of the virus in camels and to genetically characterize the circulating virus from different regions. This will be highly beneficial in the development of an appropriate vaccine.

Perborate Activated Peroxymonosulfate Process for Improving the Coagulation Efficiency of Microcystis aeruginosa by Polymeric Aluminum Chloride.

In this study, the sodium perborate (SP)-activated peroxymonosulfate (PMS) process was used to enhance the coagulation efficiency of cyanobacteria with polymeric aluminum chloride (PAC), aiming to efficiently mitigate the impact of algal blooms on the safety of drinking water production. The optimal concentrations of SP, PMS, and PAC were determined by evaluating the removal rate of OD680 and zeta potential of the algae. Experimental results demonstrated that the proposed ternary PMS/SP/PAC process achieved a remarkable OD680 removal efficiency of 95.2%, significantly surpassing those obtained from individual treatments with PMS (19.5%), SP (5.2%), and PAC (42.1%), as well as combined treatments with PMS/PAC (55.7%) and PMS/SP (28%). The synergistic effect of PMS/SP/PAC led to the enhanced aggregation of cyanobacteria cells due to a substantial reduction in their zeta potential. Flow cytometry was performed to investigate cell integrity before and after treatment with PMS/SP/PAC. Disinfection by-products (DBPs) (sodium hypochlorite disinfection) of the algae-laden water subsequent to PMS/SP/PAC treatment declined by 57.1%. Moreover, microcystin-LR was completely degraded by PMS/SP/PAC. Electron paramagnetic resonance (EPR) analysis evidenced the continuous production of SO4•-, •OH, 1O2, and O2•-, contributing to both cell destruction and organic matter degradation. This study highlighted the significant potential offered by the PMS/SP/PAC process for treating algae-laden water.