Spleen Tissues Research Articles

Toxicity studies of compound spermatogenic pill:Acute toxicity and subacute toxicity

Ethnopharmacological relevanceSpermatogenic Pill (SP) is a commonly used clinical preparation in the Third Clinical Hospital of Changchun University of Traditional Chinese Medicine. Has accumulated a good reputation for more than a decade. However, because SP is a hospital clinical agent, it has received little extensive attention from researchers, which has led to a systematic lack of basic research on it. Therefore, it is impossible to determine whether there are safety hazards that may limit its widespread clinical application, and an in-depth toxicological evaluation of SP is essential and urgent. Aim of the studyThe aim of this study was to assess the safety of SP by conducting acute and subacute toxicity examinations. Materials and methodsIdentify active compounds contained in SP by LC-MS, and determination of inorganic elements in SP using ICP-MS. The in vivo acute toxicity of SP was assessed over a duration of 14 days following administration at doses of 7.5, 15, or 30 g/kg. To evaluate subacute toxicity, mice were administered daily doses of SP (7.5, 15, or 30 g/kg) for a period of 28 days. After the treatment period, the animals were euthanized, and standard blood tests, liver and kidney function tests, as well as tests related to glycolipid metabolism, were performed. The principal organs of the mice were collected to calculate organ coefficients and undergo hematoxylin-eosin (HE) staining. ResultsLC-MS analysis showed that the active components of SP include Quercetin, Kaempferol, Beta-sitosterol, Stigmasterol, Diosbulbin B, Schizandrin, Naringenin, 2,3-hydroxycinnamic acid, L-proline, Histidine and Pluviatolide. The total amount of detected inorganic elements accounted for 3.0919% of SP. During the SP acute toxicity experiment, the groups that received the drug exhibited no signs of adverse reactions or poisoning symptoms. In subacute toxicity experiments, drug-treated mice showed overall favorable status, but the effects of continuous administration of the 30 g/kg group on body weight and food intake were reduced. An increase in the white marrow of spleen tissue after long-term administration of the drug treatment was also observed, suggesting that the drug can increase the maturation process and the number of mature lymphocytes in the spleen, and improve the lymphocyte immunity and humoral immunity function of the organism. Suggests that possibly this should be taken into account in clinical application. The routine blood examinations, as well as the assessments of liver and kidney functions, and the tests for glucose and lipid metabolism, did not reveal any notable toxic effects. ConclusionSP contains more flavonoids, and terpenoid active ingredients, and is non-toxic in the body. This discovery not only strengthens the safety foundation of its clinical application, provides a solid scientific basis for the establishment of reasonable clinical dosage and the implementation of effective clinical toxicity monitoring, but also further lays a solid theoretical cornerstone for the subsequent clinical drug trials.

Identification, genome-wide sequencing and analysis of drug resistance genes in a novel Vibrio harveyi strain isolated from yellowfin seabream

Yellowfin seabream (Acanthopagrus latus) is a fast-growing, adaptable species crucial for fish farming in southern China. However, as aquaculture of this species expands, diseases caused by bacteria, particularly Vibrio harveyi, are causing substantial economic losses. In this study, we isolated and identified a Vibrio harveyi strain denoted VH-AQ-SCAU-GZ23 from the diseased fish. Clinical signs of VH-AQ-SCAU-GZ23 infection were slow swimming, loss of appetite, loss of scales, severe liver congestion, and swelling of the gill filaments. VH-AQ-SCAU-GZ23 also induced an inflammatory response in yellowfin snapper spleen, liver, and brain tissue. To examine its pathogenicity and antibiotic susceptibility, we analyzed the genome sequence of this strain with third-generation high-throughput technology. The VH-AQ-SCAU-GZ23 genome was found to comprise a circular chromosome and three plasmids. Virulence factor analysis indicated that VH-AQ-SCAU-GZ23 contains a TLH gene encoding a thermolysin that increases virulence. Additionally, several antibiotic resistance genes were identified, including those encoding β-lactams, tetracyclines, aminoglycosides, quinolones, macrolides, peptides, and other antibiotics. To assess the drug sensitivity of those antibiotic resistance genes, we cloned ten resistance genes and transfected them into Escherichia coli DH5α for drug susceptibility testing. These resistance genes enhanced the resistance of DH5α to streptomycin, ampicillin, and penicillin G, among other antibiotics. Our study provides crucial support and a theoretical basis for advancing understanding of Vibrio harveyi. The insights gained are expected to provide guidance for future endeavors aimed at preventing and managing marine bacterial infections.

Integration of proteomics and metabolomics analysis investigate mechanism of As-induced immune injury in rat spleen

Arsenic (As) is a widespread metalloid and human carcinogen found in the natural environment, and multiple toxic effects have been shown to be associated with As exposure. As can be accumulated in the spleen, the largest peripheral lymphatic organ, and long-term exposure to As can lead to splenic injury. In this study, a Sprague-Dawley (SD) rat model of As-poisoned was established, aiming to explore the molecular mechanism of As-induced immune injury through the combined analysis of proteomics and metabolomics of rats’ spleen. After feeding the rats with As diet (50 mg/kg) for 90 days, the spleen tissue of the rats in the As-poisoned group was damaged, the level of As was significantly higher than that of the control group (P < 0.001), and the level of inflammatory cytokine interleukin-6 (IL-6) was decreased (P < 0.01). Proteomics and metabolomics results showed that a total of 134 differentially expressed proteins (DEPs) (P < 0.05 and fold change > 1.2) and 182 differentially expressed metabolites (DEMs) (VIP >1 and P < 0.05) were identified in the spleens of the As poisoned group compared to the control group (As/Ctrl). The proteomic results highlight the role of hypoxia-inducible factors (HIF), natural killer cell mediated cytotoxicity, and ribosomes. The major pathways of metabolic disruption included arachidonic acid (AA) metabolism, glycerophospholipid metabolism and folate single-carbon pool. The integrated analysis of these two omics suggested that Hmox1, Stat3, arachidonic acid, phosphatidylcholine and leukotriene B4 may play key roles in the mechanism of immune injury to the spleen by As exposure. The results indicate that As exposure can cause spleen damage in rats. Through proteomic and metabolomic analysis, the key proteins and metabolites and their associated mechanisms were obtained, which provided a basis for further understanding of the molecular mechanism of spleen immune damage caused by As exposure.

Integrated transcriptomics and metabolomics reveal the mechanism of polystyrene nanoplastics toxicity to mice

Microplastic (MP) are an emerging environmental pollutant, which has toxic effects on organisms, and it has received extensive attention currently. Studying the transcriptomic and metabolic responses of mice to nanoplastic-contaminated water is critical for understanding molecular-level toxicity of nanoplastics (NPs), but there are few studies on this topic. To analyze the effects of different concentrations of polystyrene (PS) nanoplastic-contaminated water on mice at the transcriptome and metabolism of spleens to study the molecular toxicity. Here, testing of histopathology of spleen of female mice was performed after drinking water containing 0.1 μm PS-NPs (1 mg/mL and 50 mg/mL) at different concentrations for 49 days, respectively. The spleen tissue samples were subjected to metabolome and transcriptome sequencing. Four differentially expressed genes were randomly chosen for qRT-PCR to confirm the correctness of transcriptome sequencing. Common Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that a large number of differential genes and differential metabolites mainly focused on immune, inflammation, neurodegenerative disease, cardiovascular disease, nervous, etc. in the organism systems module; lipid, amino acid, taurine and hypotaurine metabolisms, etc. in the metabolism module; signaling translation, signaling molecules and interaction, and neuroactive ligand-receptor interaction, etc. in the environmental information processing. The results showed that pathway analysis at transcriptome and metabolome levels confirmed that the immune system of mice was affected after drinking water contaminated with polystyrene nanoplastics.

Identification of immune-associated genes with altered expression in the spleen of mice enriched with probiotic Lactobacillus species using rna-seq profiling.

Probiotics are living microorganisms that can provide health benefits when consumed. Here, we investigated the effects of probiotics on gene expression in the spleen of mice using RNA-sequencing analysis between negative control and probiotic groups (including 4 Lactobacillus strains: Lactobacillus fermentum, L. casei, L. plantarum, and L. brevis). Mice exposed with probiotic in 4 weeks by intragastric administration. Then, spleen tissues of the control and probiotics groups were collected on days 14 and 28 for RNA sequencing. In total, 665, 186, and 81 DEGs were significantly expressed on day 14 vs. control, day 28 vs. control groups, and probiotics day 28 vs. day 14 groups, respectively. On the other hand, 12 Toll-like receptor (TLR) genes underwent additional validation through qRT-PCR, affirming the increased alignment between qRT-PCR and RNA-Seq findings. In addition, the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses revealed that the DEGs were predominantly enriched in defense responses to pathogens, including inflammatory bowel diseases, malaria, leukaemia virus 1, and herpes virus, as well as immune processes related to immune response and signal transduction. This study represents the first investigation into mice's gene expression in the spleen exposed to probiotics using Lactobacillus spp. isolated from a field strain in Vietnam. Our results provide valuable insights into the impacts and functions of probiotics on mammalian development, offering crucial information for the potential therapeutic use of probiotics in defending against pathogens in Vietnam. The findings from this study highlight the potential of probiotics in modulating gene expression in the spleen, which may have implications for immune function and overall health in mice.

Peptidomic profiling of endogenous peptides in the spleens of mouse models of Graves' disease

BackgroundGraves' disease (GD) is a common autoimmune thyroid disorder. The pathogenesis of GD involves an autoimmune response to the A subunit of the human thyrotropin receptor (hTSHR), although the specific mechanisms are not fully elucidated. MethodsThis study established a GD model by immunizing BALB/c mice with a recombinant adenovirus expressing the hTSHR A subunit. Spleen tissues were collected and analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify differentially expressed peptides (DEPs). Gene Ontology (GO) analysis and KEGG pathway analysis were further utilized to annotate the functions of these DEPs. Additionally, peptide bioactivity prediction and molecular docking studies were conducted using Alphafold and Pymol software, respectively, to assess the binding affinity of specific peptides to the hTSHR A subunit. ResultsThe GD mouse model was successfully established, and 1,428 DEPs were identified in the spleen, with 368 upregulated and 1,060 downregulated. Functional analysis indicated that these DEPs are mainly involved in cellular endocytosis, regulation of gene expression, and nucleocytoplasmic transport. Notably, molecular docking studies revealed that the abnormally highly expressed peptide HG2A-24aa demonstrated potential bioactivity and strong binding affinity with hTSHR-289aa. ConclusionThe specific bioactive peptides may play key roles in the pathogenesis of GD, particularly HG2A-24aa, which may have a significant role in the MHC II antigen presentation pathway mediated by the hTSHR A subunit.

Effect of rock bream iridovirus (RBIV) contained tissue intake on rock bream (Oplegnathus fasciatus) mortality and blood cell distribution

Rock bream (Oplegnathus fasciatus) is one of the highly priced cultured marine fish in Korea. Rock bream iridovirus (RBIV) outbreaks in aquaculture farms may involve environmental factors, co-infection with other pathogenic microorganisms and grounded (raw) fish feed. This study evaluated the effects of RBIV-containing tissue intake on mortality and oral transmission in rock bream. Virus-containing tissues administered to rock bream [50 mg (1.53 × 108/major capsid protein, MCP gene copies) to 2400 mg (7.34 × 109)] held at 23 °C lead to 100 % mortality by 27 days post administration. Interestingly, the mortality rates were not viral dose- or concentration dependent. Further, high MCP gene copy numbers were observed in the gill, liver, intestine, stomach, spleen, heart, kidney, brain and muscle tissues (viral load range of 3.03 × 106 to 4.01 × 107/mg, average viral load 1.70 × 107/mg) of dead rock bream. Moreover, a high viral load was detected in the intestine and stomach, where the virus was directly administered. This indicated that the intake of RBIV-containing tissue feed weakens the intestinal mucosal immunity and increases viral load in the intestine. Moreover, the levels of complete blood cell count (CBC) indicators, such as red blood cell (RBC), hemoglobin (HGB) and hematocrit (HCT) significantly decreased from 15 dpi with red blood cell distribution width (RDW), and white blood cells (lymphocyte, monocyte and granulocyte) significantly increased from the initial to later stage of infection. These results highlight the significance of blood-mediated indicators against RBIV infection in rock bream. We demonstrate the existence of an oral transmission route for RBIV in rock bream. Our findings indicate that pathogen-containing feed is an important risk factor for disease outbreaks in rock bream.

Identification and phylogenetic analysis of bovine herpesvirus 4 isolated from aborted bovine foetuses in Türkiye.

Bovine herpesvirus 4 (BoHV-4) is responsible for reproductive disorders and abortions in cattle. The aim of this study was to investigate the potential role of BoHV-4 in abortions in dairy cows when the possible aetiological cause could not be determined in Türkiye. The study also provided the molecular characterization of BoHV-4 strains based on the gB gene. In this study, foetuses (n = 188) and serum (n = 50) were retrospectively used for PCR and indirect ELISA assays, respectively. In addition, foetuses with BoHV-4 were tested for the isolation of the BoHV-4 strain in MDBK cell culture. The BoHV-4 gB gene was detected in the brain, liver, lung, kidney, and spleen tissues of three foetuses out of 188 aborted foetuses. The BoHV-4 strain was isolated from the kidney of one foetus in MDBK cell culture. The three aborted foetuses were from different cows in the same herd in 2021. On this dairy farm, the BoHV-4 seroprevalence rate in cattle with a history of abortion was 96% (48/50). Phylogenetic analysis revealed that the detected strains belong to genotype II. The present study shows that the impact of BoHV-4 on abortions in cattle should not be ignored. It can therefore be informative to focus on BoHV-4 in routine laboratory diagnostics for abortions without a proven aetiology or in cattle herds that have previously been infected with BoHV-4.

Relationship between mean monthly February temperature and the level of oxidative processes in immunocompetent organs of bream of Rybinsk reservoir in different years

The intensity of oxidative processes in immunocompetent organs of immature bream Abramis brama L. with an average weight of 375±30 g and length of 290±20 cm was studied. The fish were caught with shuttle nets in the Volga Plec of the Rybinsk reservoir in different years in terms of average daily temperature conditions. After autopsy, samples of immunocompetent organs (kidney, spleen and liver) were taken from captured fish into sterile tubes. Samples were then frozen and stored in a freezer at minus 18–20 °C until processing time. In laboratory conditions, samples were thawed at room temperature immediately before analysis. For further studies, homogenate was prepared from kidney, spleen and liver tissues using a homogenizer with physiological solution in the ratio of 1:1. In the homogenates the content of lipid peroxidation products was studied by the content of malonic dialdehyde and the level of antioxidant defense by the index of inhibition of substrate oxidation. The interannual variability of the studied parameters was noted, associated with ecological and climatic conditions, in particular, with temperature. The most significant differences of the studied biochemical parameters were revealed in the year with the highest temperature conditions compared to the year with the lowest temperature. The lowest values of malonic dialdehyde and substrate oxidation constant were observed in fish in the year with the lowest average daily temperature. In warmer year intensification of lipid peroxidation processes and decrease of antioxidant defense level in the studied organs were observed. The results obtained can be used in monitoring the impact of climatic factors on the physiological and biochemical status of fish.

Casein phosphopeptide/antimicrobial peptide co-modified SrTiO3 nanotubes for prevention of bacterial infections and repair of bone defects

Endowing titanium surfaces with multifunctional properties can reduce implant-related infections and enhance osseointegration. In this study, titanium dioxide nanotubes with strontium doping (STN) were first created on the titanium surface using anodic oxidation and hydrothermal synthesis techniques. Next, casein phosphopeptide (CCP) and an antimicrobial peptide (HHC36) were loaded into the STN with the aid of vacuum physical adsorption (STN-CP-H), giving the titanium surface a dual function of "antimicrobial-osteogenic". The surface of STN-CP-H has a suitable roughness and good hydrophilicity, which is conducive to osteoblasts. STN-CP-H had a 99 % antibacterial rate against S. aureus and E. coli and effectively prevented the growth of bacterial biofilm. Meanwhile, the antibacterial mechanism of STN-CP-H was initially explored with the help of transcriptome sequencing technology. STN-CP-H could greatly increase osteoblast adhesion, proliferation, and expression of osteogenic markers (alkaline phosphatase, runt-related transcription) when CCP and Sr worked together synergistically. In vivo, the STN-CP-H coating could effectively promote new osteogenesis around titanium implant bone and had no toxic effects on heart, liver, spleen, lung and kidney tissues. A potential anti-infection bone healing material, STN-CP-H bifunctional coating developed in this work efficiently inhibited bacterial infection of titanium implants and encouraged early osseointegration.

A cross-sectional study to correlate nuclear fixative properties of formal saline and clarke’s fluid for histomorphology evaluation of human tissues

Abstract Background: Formal saline is an extensively used cheapest and easily available tissue fixative. However, formalin has been proven to have carcinogenic effects. Clarke’s fluid is an alternative to formal saline and is effective in maintaining the morphological and cellular architecture. Objectives: The objective of the study was to compare the nuclear fixative properties of Clarke’s fluid and formal saline. Materials and Methods: The fresh human tissues of thin skin, large-sized arteries, lymph nodes, spleen, and ileum were obtained from the fresh cadaver. The tissues were fixed in two different fixatives namely 10% formalin and Clarke’s fluid. Then, the tissues were histologically processed. The hematoxylin and eosin-stained serial sections of tissues were graded based on the ease of microtomy, cytoplasmic, and nuclear staining scores. Results: The nuclear properties of all the tissues immersed in Clarke’s fluid showed a median score of 3 and Grade 2 for formal saline; cytoplasmic properties for spleen and skin were median scores of Grade 3 for Clarke’s fluid. The artery was graded 3 for both the fixatives. The Kruskal–Wallis test when applied to nuclear properties, cytoplasmic adequacy, and tissue shrinkage for tissues stained with Clarke’s fluid was statistically significant (P &lt; 0.01). This inferred that Clarke’s fluid is a better nuclear fixative than formal saline. Conclusion: The imminent risks associated with formal saline paved us to explore substitute fixatives, which could provide better results and safety for laboratory workers. Clarke’s fluid is an excellent substitute for formal saline as a nuclear fixative.

Lentiviral vectors for precise expression to treat X-linked lymphoproliferative disease

X-linked lymphoproliferative disease (XLP1) results from SH2D1A gene mutations affecting the SLAM-associated protein (SAP). A regulated lentiviral vector (LV), XLP-SMART LV, designed to express SAP at therapeutic levels in T, NK, and NKT cells, is crucial for effective gene therapy. We experimentally identified 34 genomic regulatory elements of the SH2D1A gene and designed XLP-SMART LVs to emulate the lineage and stage-specific control of SAP. We screened them for their on-target enhancer activity in T, NK, and NKT cells and their off-target enhancer activity in B cell and myeloid populations. In combination, three enhancer elements increased SAP promoter expression up to 4-fold in on-target populations invitro. NSG-Tg(Hu-IL15) xenograft studies with XLP-SMART LVs demonstrated up to 7-fold greater expression in on-target cells over a control EFS-LV, with no off-target expression. The XLP-SMART LVs exhibited stage-specific T and NK cell expression in peripheral blood, bone marrow, spleen, and thymic tissues (mimicking expression patterns of SAP). Transduction of XLP1 patient CD8+ Tcells or BM CD34+ cells with XLP-SMART LVs restored restimulation-induced cell death and NK cytotoxicity to wild-type levels, respectively. These data demonstrate that it is feasible to create a lineage and stage-specific LV to restore the XLP1 phenotype by gene therapy.

Spleen Swabs for Sensitive and High-Throughput Detection of African Swine Fever Virus by Real-Time PCR.

African swine fever (ASF) continues to spread in Africa, Europe, Asia and the island of Hispaniola, increasing the need to develop more streamlined and highly efficient surveillance and diagnostic capabilities. One way to achieve this is by further optimization of already established standard operating procedures to remove bottlenecks for high-throughput screening. Real-time polymerase chain reaction (real-time PCR) is the most sensitive and specific assay available for the early detection of the ASF virus (ASFV) genome, but it requires high-quality nucleic acid extracted from the samples. Whole blood from live pigs and spleen tissue from dead pigs are the preferred samples for real-time PCR. Whole blood can be used as is in nucleic acid extractions, but spleen tissues require an additional homogenization step. In this study, we compared the homogenates and swabs prepared from 52 spleen samples collected from pigs experimentally inoculated with highly and moderately virulent ASF virus strains. The results show that not only are the spleen swabs more sensitive when executed with a low-cell-count nucleic acid extraction procedure followed by real-time PCR assays but they also increase the ability to isolate ASFV from positive spleen samples. Swabbing is a convenient, simpler and less time-consuming alternative to tissue homogenization. Hence, we recommend spleen swabs over tissue homogenates for high-throughput detection of ASFV by real-time PCR.

The Protective Effect of on The Reproductive System and Some Visceral Organs (Liver, Spleen) Tissues of Female Rats Exposed to High Dose of Zinc Sulphate in Drinking Water

The Protective Effect of on The Reproductive System and Some Visceral Organs (Liver, Spleen) Tissues of Female Rats Exposed to High Dose of Zinc Sulphate in Drinking Water

Very virulent infectious bursal disease virus infection triggered microscopic changes, apoptosis, and inflammatory cytokines imbalance in chicken spleen and thymus

ABSTRACT Infectious bursal disease virus (IBDV) can cause a highly contagious disease, resulting in severe damage to the immune system that causes immunosuppression in young chickens. Both spleen and thymus are important immune organs, which play a key role in eliciting protective immune responses. However, the effects of very virulent IBDV (vvIBDV) strain LJ-5 infection on chicken spleen and thymus are still unknown. In the present study, 3-week-old specific pathogen-free chickens were infected with vvIBDV for 1–5 days. The vvIBDV infection significantly increased the spleen index and decreased the thymus index. Microscopic analysis indicated necrosis, depletion of the lymphoid cells, and complete loss of structural integrity in spleen and thymus. Ultrastructural analysis displayed mitochondrial and nuclear damage, including mitochondrial cristae breaks, and deformation of nuclear membrane in vvIBDV-infected spleen and thymus tissues. Cytokine levels increased in the spleen and thymus after IBDV infection, promoting inflammation and causing an inflammatory imbalance. Moreover, the mRNA expression of apoptosis-related genes was significantly upregulated in the vvIBDV-infected group compared to the control group. Meanwhile, the mRNA expression of mitochondrial dynamics was altered in the spleen and thymus of vvIBDV-infected chickens. These results suggested that vvIBDV infection triggers an imbalance of inflammatory cytokines, and apoptosis in the spleen and thymus, resulting in immune injury in chickens. This study provides basic data for the further study of vvIBDV pathogenesis.

Prenatal exposure to bisphenol AF causes toxicities in liver, spleen, and kidney tissues of SD rats

As a replacement for bisphenol A (BPA), bisphenol AF (BPAF) showed stronger maternal transfer and higher fetal accumulation than BPA. Therefore, concerns should be raised about the health risks of maternal exposure to BPAF during gestation on the offspring. In this study, SD rats were exposed to BPAF (0, 50, and 100 mg/kg/day) during gestation to investigate the bioaccumulation and adverse effects in liver, spleen, and kidney tissues of the offspring at weaning period. Bioaccumulation of BPAF in these tissues with concentrations ranging from 1.56 ng/mg (in spleen of males) to 55.44 ng/mg (in liver of females) led to adverse effects at different biological levels, including increased relative weights of spleen and kidneys, histopathological damage in liver, spleen, and kidney, organ functional damage in liver, spleen, and kidney, upregulated expression of genes related to lipid metabolism (in liver), oxidative stress response (in kidney), immunity and inflammatory (in spleen). Furthermore, dysregulated metabolomics was identified in spleen, with 217 differential metabolites screened and 9 KEGG pathways significantly enriched. This study provides a comprehensive insight into the systemic toxicities of prenatal exposure to BPAF in SD rats. Given the broad applications and widespread occurrence of BPAF, its safety should be re-considered.

A comprehensive study on the multifunctional properties of galectin-4 in red-lip mullet (Planiliza haematocheilus): Insights into molecular interactions, antimicrobial defense, and cell proliferation

Galectin-4 belongs to the galactoside-binding protein family and is a type of tandem repeat galectin. Despite previous studies indicating its importance in fish immunology, a comprehensive investigation is necessary to fully understand its role in immunomodulatory functions and cellular dynamics. Therefore, this study aimed to explore the immunomodulatory functions of galectin-4 with a particular focus on its antimicrobial and cellular proliferative properties. The open reading frame of PhGal4 spans 1092 base pairs and encodes a soluble protein of 363 amino acids with a theoretical isoelectric point (IEP) of 6.39 and a molecular weight of 39.411 kDa. Spatial expression analysis under normal physiological conditions revealed ubiquitous expression of PhGal4 across all examined tissues, with the highest level observed in intestinal tissue. Upon stimulation with poly I:C, LPS, and L. garvieae, a significant increase (p < 0.05) in PhGal4 expression was observed in both blood and spleen tissues. Subsequent subcellular localization assay demonstrated that PhGal4 was predominantly localized in the cytoplasm. The recombinant PhGal4 (rPhGal4) exhibited specific binding capabilities to pathogen-associated molecular patterns (PAMPs), including LPS and peptidoglycan, but not poly I:C. The rPhGal4 negatively affected the bacterial growth kinetics. Additionally, rPhGal4 demonstrated complete hemagglutination of fish erythrocytes, which could be inhibited by the presence of D-galactose and α-lactose. The overexpression of PhGal4 in FHM epithelial cells demonstrated a significant suppression of viral replication during VHSV infection. Furthermore, the in vitro scratch assay and WST-1 assay demonstrated a wound healing effect of PhGal4 overexpression in FHM cells, potentially achieved through the promotion of cell proliferation by activating genes involved in cell cycle regulation. In conclusion, the responsive expression to immune stimuli, antimicrobial properties, and cell proliferation promotion of PhGal4 suggest that it plays a crucial role in immunomodulation and cellular dynamics of red-lip mullet. The findings in this study shed light on the multifunctional nature of galectin-4 in teleost fish.

Spontaneous cytokine production by the cells from peripheral blood, lymph nodes, spleen, thymus and skin in a murine model of acute psoriasis-like dermatitis

Imiquimod-induced model of psoriasis in mice is used for the studies in psoriasis. Imiquimod administered as skin application triggers a complex multi-stage process simulating psoriatic inflammation of the skin, accompanied by changes in production of cytokines. The aim of this work was a comprehensive study of their contents in cell supernatants isolated from skin, blood, from central and peripheral lymphoid organs and the effect of imiquimod on these parameters. Forty-six C57BL/6 mice were divided into two groups: (1) control (n = 22), and (2) experimental (n = 24). Development of experimental pathology was evaluated by van de Fits et al. The mice from experimental group were treated with a cream containing 5% imiquimod (62.5 mg/ cm2/ day/mouse) for 7 days. On the 7th day of experiment, the animals were subject to blood sampling, extraction of spleen, lymph nodes, thymus, and skin biopsies were also made. To obtain cell suspensions, the tissues of spleen, lymph nodes, and thymus were crushed in a homogenizer, and then passed through cell filters (50 mcm pore size). The method of spontaneous migration was used to isolate skin cells. The cultures of blood, spleen, lymph nodes, thymus, and skin cells were incubated for 24 hours in RPMI-1640 followed by assessment of spontaneous cytokine production in supernatants. The cytokine level (IFNγ, IL-1β, IL-5, IL-6, IL-10, IL- 17, TNFα, IL-10) was determined using the murine Th1/Th2/Th17 Panel test system. Our study revealed that the cells isolated from blood and lymphoid organs synthesize increased amounts of pro-inflammatory cytokines: IL-1 and IL-17. The most pronounced changes in the cytokine profile are observed in cell supernatants from blood, skin and spleen cultures.

Development of a peptide-generated antibody to rabbit hemorrhagic disease virus 2 VP60 and its immunohistochemical application in natural cases.

Rabbit hemorrhagic disease virus 2 (RHDV2) has spread across the United States infecting and causing death in domestic and wild rabbits. Immunohistochemistry (IHC) would be a useful tool for the detection of RHDV2 antigen in tissues as it is inexpensive and readily achievable in most diagnostic laboratories. However, there is no readily available antibody for this purpose. To fill this void, we generated an RHDV2 capsid protein VP60-specific antibody in chicken eggs and validated the antibody using formalin-fixed tissues from 5 domestic rabbits naturally infected with RHDV2. Viral antigen was detected immunohistochemically in various tissues, most prominently in hepatocytes and macrophages in liver, and in macrophages in spleen and cecal lymphoid tissue. Intravascular mononuclear cells in lung and renal tubular and biliary epithelium also were immunolabeled. Both nuclear and cytoplasmic immunolabeling were observed. This peptide-generated antibody is a potentially useful tool as an adjunct to reverse-transcription PCR or in situ hybridization for detection of RHDV2 in tissues.

Do microplastics accumulate in penguin internal organs? Evidence from Svenner island, Antarctica

The prevalence of microplastics (MPs, <5 mm) in natural environments presents a formidable global environmental threat MPs can be found from the Arctic to Antarctica, including glaciers. Despite their widespread distribution, studies on MP accumulation in apex predators inhabiting Polar Regions remain limited. The objective of this study was to conduct a comprehensive examination, for the first time, of MP bioaccumulation in various organs and tissue of Adélie penguins. This investigation comprehends the gastrointestinal tract (GIT), scat, internal organ (lung, trachea, spleen, and liver) and tissue (muscle) samples collected from Svenner Island, Antarctica during the 39th Indian expedition to Antarctica in 2019–2020. Our analyses revealed the presence of 34 MPs across the GIT, scat, lung, and trachea samples, with no MPs detected in muscle, spleen, or liver tissues. Blue-colored microfibers (>50 %) and MPs smaller than 1 mm (38 %) in size were prominently observed. Polymer characterization utilizing μ-FTIR spectroscopy identified low-density polyethylene (LDPE) (~63 %) as the predominant polymer type. The accumulation of MP fibers in the gastrointestinal tract and scat of Adélie penguins may originate from marine ambient media and prey organisms. Furthermore, the presence of LDPE fibers in the trachea and lungs likely occurred through inhalation and subsequent deposition of MPs originating from both local and long-range airborne sources. The identification of fibers ranging between 20 and 100 μm within the trachea suggests a plausible chance of cellular deposition of MPs. Overall our findings provide valuable insights into the organ-specific accumulation of MPs in apex predators. Adélie penguins emerge as promising environmental bio-monitoring species, offering insights into the potential trophic transfer of MPs within frigid environments.