Search Results (53)

Spinal muscular atrophy (SMA) is a neuromuscular disorder caused by loss of the SMN1 gene, reduced levels of SMN protein, and motor neuron degeneration. However, increasing evidence shows that SMA is a multisystemic disease with immune system involvement. We investigated how SMN deficiency affects lymphoid organ development and function using a severe SMA mouse model (SMNΔ7) and postmortem human fetal and postnatal tissues lacking SMN1 and carrying one or two SMN2 copies, consistent with type 0–I SMA. Histology, immunostaining, and flow cytometry were used to examine tissue architecture and immune cell composition. SMNΔ7 mice displayed thymus, spleen, and bone marrow abnormalities, including mislocalization of T- and B-cells and expansion of resident macrophages. Bone marrow analysis revealed impaired B-cell development, suggesting intrinsic hematopoietic defects rather than apoptosis. Early treatment with a nusinersen-like antisense oligonucleotide, administered intracerebroventricularly or subcutaneously, restored SMN2 splicing, improved survival, motor function, and prevented lymphoid pathology. Human SMA samples exhibited similar, though milder, splenic alterations compared to SMNΔ7 mice, while thymic organization remained largely preserved. These findings demonstrate that SMN deficiency disrupts lymphoid organ development through defective bone marrow output and impaired immune cell maturation. Early SMN restoration prevents these abnormalities, highlighting immune dysfunction as a key component of SMA pathology. Full article

Background: The positive identification of a source of tissue as human plays an important role in various contexts. It is particularly important for investigations concerning tissue and organ trafficking, since unequivocal confirmation is required for legal proceedings involving such cases. While deoxyribonucleic (DNA) methods are considered the gold standard for tissue identification, issues such as degraded DNA or the presence of chemical preservatives can hinder performance and positive identification using DNA techniques. Objectives: The aim of this study was to develop a simple method for presumptive identification of human tissue using standard bottom-up proteomics data. Methods: We identified proteins isolated from human kidney, lung and spleen tissues by bottom-up proteomics and database search using Proteome Discoverer and Sequest HT algorithms. The list of identified proteins was sorted based on liquid chromatography (LC)–mass spectrometry (MS) data metrics such as the number of unique peptides used to identify each protein and the % sequence coverage of an identified protein to determine if any parameter would cluster proteins annotated as human in a distinct category. We found that eliminating proteins identified with fewer than two unique peptides and those with less than 5% sequence coverage resulted in a final list where at least half of the remaining proteins are annotated as human. We applied this data filtration process to blinded LC–MS/MS data from 26 previous experiments to assess accuracy. Results: Using bottom-up proteomics data and the filtration rules established, we identified tissue samples (n = 10), including kidney, spleen, lung, formalin-fixed paraffin-embedded uterus, frozen breast tissue, dry blood and dry saliva as human, and tissue (n = 16) from rat, mouse, bovine, and sheep as non-human, resulting in 100% sensitivity and specificity. Conclusions: The results demonstrate that the list of identified proteins following a standard bottom-up proteomics experiment could be filtered and potentially used as a fast and simple method for presumptive human tissue identification. Full article

The use of phytobiotics in aquafeeds is a promising strategy to enhance performance and resilience to disease. This study evaluated the protective role of Morus alba (MA) extract against Vibrio cholerae, integrating in vivo responses in Dormitator latifrons (growth, biochemical and enzymatic responses, haemato-immunology and tissue histopathology) with in vitro assessment of V. cholerae growth, virulence-associated gene expression and cellular morphology. D. latifrons juveniles were fed five diets (0, 5, 10, 15 and 20 g/kg feed; three tanks per treatment, 15 fish per tank) for eight weeks, followed by a 7-day challenge with V. cholerae. MA increased growth and feed utilisation (p < 0.05); the 20 g/kg group reached 27.57 g final weight with a feed conversion ratio of 1.24, and whole-body protein and lipid contents rose at higher doses. MA modulated plasma biochemistry and key digestive (amylase, lipase), metabolic (ALT, AST) and antioxidant (SOD, CAT, GPx) enzymes, and improved haematological profiles. Histology of the intestine, liver and spleen showed preserved architecture and reinforced mucosal features in supplemented fish, particularly at 15–20 g/kg. Post-challenge, supplemented groups exhibited higher survival/relative protection than controls, alongside lower transaminases and stronger antioxidant responses. In vitro, MA extract inhibited V. cholerae growth, attenuated virulence-associated gene (toxR, ompU) expression and induced marked morphological damage in planktonic cells. Multivariate analyses (Z-score heatmaps and PCA) linked immune–enzymatic improvements with growth and protection. Overall, 15–20 g/kg MA optimised immunophysiological status and disease resistance, supporting MA as a functional feed additive for sustainable aquaculture of D. latifrons. Full article

In China, bites caused by the Naja atra and Deinagkistrodona acutus are the most common types of snakebites. While the functional characteristics of the two venom components have been well documented, their in vivo metabolic pathways, target organ distribution patterns, and dynamic pharmacokinetic profiles remain less explored. This study established a murine envenoming model through CY7-SE labeling of Naja atra and Deinagkistrodon acutus venoms. The real-time in vivo absorption and biodistribution of venoms were dynamically monitored via fluorescence imaging, with subsequent proteomic profiling to characterize organ-specific toxin targeting patterns. Gel filtration chromatography and HPLC analyses validated labeling efficiency at ratios of 0.1 mg CY7-SE per 1 mg Naja atra venom and 0.075 mg CY7-SE per 1 mg Deinagkistrodon acutus venom, with electrophoretic confirmation of protein integrity and preserved 740 nm fluorescence excitation. Acute toxicity assays demonstrated no significant difference in LD₅₀ lethality between labeled and native venoms (p > 0.05). The intoxication models revealed species-specific pathophenotypes, i.e., CY7-Naja atra venom induced systemic weakness, tachypnea, and inflammatory necrosis in lung, myocardium, and liver, whereas CY7-Deinagkistrodon acutus venom provoked hemorrhagic diathesis. Both models exhibited marked leukocytosis, transaminitis, and elevated creatinine levels (p < 0.05). Fluorescence tracing uncovered distinct biodistribution kinetics: Deinagkistrodon acutus venom achieved peak organ accumulation at 3 h with rapid dissemination (24 h injection-site retention: 12.61%), contrasting with Naja atra venom’s delayed 6 h peak and prolonged renal sequestration (24 h injection-site retention: 60.9%). Target organ proteomic profiling identified Deinagkistrodon acutus-enriched thrombin-like enzymes and metalloproteinases in lung/liver/spleen, while Naja atra venom predominantly accumulated renal acidic phospholipase A₂ and weakly neurotoxic NNAM2. Full article

Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by joint inflammation that leads to tissue damage and disability. RA affects approximately 0.5–1% of the global population and is driven by a complex interplay of genetic susceptibility, environmental factors, and immune dysregulation. While biologic and targeted synthetic DMARDs improved RA treatment, they have limitations in efficacy, safety, and accessibility. B-cell-targeting therapies, such as anti-CD20, have shown effectiveness, but only with broad immunosuppression, which can increase infection risk and compromise humoral immunity. Therefore, there is an unmet need for more selective therapeutic strategies that modulate pathogenic immune pathways while preserving protective immune functions. It has been suggested that targeting the BAFF pathway may offer a more favorable therapeutic approach compared to targeting CD20. Objectives: In this study, we evaluated the therapeutic potential of V3-46s mIgG2a, an anti-BAFF-R (BR3) antibody in a mouse RA model, hypothesizing that it would offer a more selective and effective strategy. Methods: We expressed and purified four antibody variants and assessed their binding and neutralizing activity in vitro. V3-46s mIgG2a was selected for in vivo evaluation in a collagen-induced arthritis (CIA) model. Results: Treatment with this antibody delayed disease onset and reduced arthritis severity, spleen index, and B-cell populations. Conclusions: These findings highlight the potential of BAFF-R-targeting antibodies as a therapeutic approach for RA treatment. This preclinical work lays the groundwork for future development of BAFF-R blockade as a complementary or alternative strategy to current biologic treatments. Full article

Breast milk is a major source of probiotics, particularly lactic acid bacteria (LAB), which are known to regulate the intestinal microbial community and exert antibacterial effects. However, little is known about the preventive effects of feline milk-derived LAB against Salmonella infection in vivo. In this study, a strain of Pediococcus acidilactici (M22) was isolated from feline milk and evaluated for its protective potential in C57BL/6 mice challenged with Salmonella Typhimurium SL1344 (VNP20009). Following oral administration of M22, mice were infected with S. Typhimurium, and protective efficacy was assessed through body weight changes, bacterial loads in tissues, histopathological examination of the colon, oxidative stress markers, cytokine profiles, and 16S rRNA gene sequencing of cecal microbiota. The results showed that pretreatment with M22 significantly reduced bacterial loads in the liver, spleen, and cecum compared with controls. M22 administration enhanced antioxidant capacity, alleviated infection-induced inflammation, and preserved intestinal barrier integrity by restoring villus morphology and upregulating tight junction proteins (ZO-1 and occludin). Microbiota analysis further revealed that M22 enriched short-chain fatty acid-producing beneficial taxa (e.g., lactic acid bacteria) while suppressing pro-inflammatory genera. Collectively, these findings provide scientific evidence that feline milk-derived P. acidilactici M22 is a safe and effective probiotic candidate. By enhancing gut health and host resistance to infection, M22 offers a promising strategy to improve companion animal health, reduce reliance on antibiotics, and mitigate zoonotic transmission of pathogens. Full article

Background: ErisNet, a novel AI model to reduce noise in Computed Tomography images. Methods: We trained ErisNet on 23 post-mortem whole-body CT scans. We assessed the objective performance with mean square error (MSE), peak signal-to-noise ratio (PSNR), structural similarity index (SSIM) measure, visual information fidelity (VIF), edge preservation index (EPI) and noise variance (NV). We assessed the qualitative performance by six radiologists. To support the visual assessment, we placed circular regions of interest (ROI) in the vitreous body, brain, liver and spleen parenchyma and paravertebral muscle. Results: ErisNet achieved MSE 64.07 ± 46.81, PSNR 31.32 ± 3.69 dB, SSIM 0.93 ± 0.06, VIF 0.49 ± 0.09, EPI 0.97 ± 0.01 and NV 64.69 ± 46.80. The ROI analysis showed a reduction in noise: the SD of the HU decreased by 8% in the vitreous body (from 17.6 to 16.2 HU), by 18% in the brain parenchyma (from 18.85 to 15.40 HU) and by 15–19% in the liver, spleen and paravertebral muscle. The six radiologists confirmed these results by assigning high scores (scale from one to five): overall quality 4.5 ± 0.6, noise suppression/detail preservation 4.7 ± 0.5 and diagnostic confidence 4.8 ± 0.4 (p < 0.01). Conclusions: ErisNet improves the quality of CT images and shows strong potential for processing low-dose scans. Full article

Background: Splenic arterial embolization (SAE) is a well-established technique in the non-operative management of splenic trauma and aneurysms. While its short-term safety and efficacy have been widely documented, medium- and long-term impacts on splenic volume and function remain under-investigated. This study aimed to evaluate volumetric changes and hematological parameters following SAE, with emphasis on its role in preserving splenic integrity and potential integration with AI-enhanced imaging technologies. Methods: We retrospectively analyzed 17 patients treated with SAE between January 2014 and December 2023. Volumetric measurements were performed using computed tomography (CT) with 3D reconstructions before and after SAE. Patients were divided into two groups based on indication: polytrauma (n = 8) and splenic artery aneurysm (n = 9). Hematological parameters including white blood cells (WBCs), red blood cells (RBCs), and hemoglobin (Hb) were evaluated in correlation with clinical outcomes. Statistical significance was assessed using Student’s t-test, and power analysis was conducted. Results: Among the trauma group, mean splenic volume decreased from 190.5 ± 51.2 cm³ to 147.8 ± 77.8 cm³ (p = 0.2158), while in the aneurysm group, volume decreased from 195.4 ± 78.9 cm³ to 143.7 ± 81.4 cm³ (p = 0.184). Though not statistically significant, these changes suggest post-procedural splenic remodeling. The technical success of SAE was 100%, with no cases of late follow-up infarction, abscess, immunological impairment, or secondary splenectomy required. Hematologic parameters remained within normal limits in follow-up assessments. Conclusions: SAE represents a safe and effective intervention for spleen preservation in both traumatic and aneurysmal conditions. Although a reduction in splenic volume has been observed, white blood cell counts, a reliable indicator of splenic function, have remained stable over time. This finding supports the preservation of splenic function following SAE. Full article

Background/Objectives: Visceral leishmaniasis remains a serious public health issue, with current antimonial therapies limited by systemic toxicity. Liposomal drug delivery systems offer a promising strategy to improve therapeutic outcomes. In this context, we previously demonstrated the antileishmanial efficacy of the liposomal formulation containing tartar emetic (Lip-TE) in reducing the parasite load in the spleen and liver of animals experimentally infected with Leishmania (Leishmania) infantum, this being an interesting alternative for the treatment of visceral leishmaniasis. In this study, we aimed to evaluate the physicochemical properties, biodistribution, pharmacokinetics, and cardiotoxicity of Lip-TE. Methods: Both Lip-Blank and Lip-TE exhibited monodisperse size distributions (PDI < 0.3), with Lip-TE having an increased particle size (205.1 ± 4.5 nm) compared to Lip-Blank (137.5 ± 0.8 nm). Both formulations demonstrated a positive zeta potential of about +17.0 mV. Biodistribution and cardiotoxicity studies were performed using female BALB/c mice, approximately 8 to 10 weeks old. Results: A significantly greater splenic, hepatic, and renal accumulation of Lip-TE was observed, particularly at 120 min post-injection, compared to Free TE treatment. The pharmacokinetic profile of the treatment with Lip-TE showed higher systemic exposure (AUC), extended half-life and mean residence time, and reduced clearance compared to Free TE treatment. Echocardiographic analysis revealed that treatment with Free TE significantly reduced the ejection fraction percentage and QRS complex prolongation after 10 days of treatment. These alterations persisted at 30 days, indicating progressive cardiotoxicity. In contrast, Lip-TE treatment preserved cardiac function, revealing the protective effect of the liposomal encapsulation. Conclusions: These findings highlight the potential of the Lip-TE treatment to enhance safety and efficacy. Full article

Hypothermia-related deaths present significant diagnostic challenges due to non-specific and often inconsistent autopsy findings. This study investigated the histological and immunohistochemical alterations associated with primary and secondary hypothermia in an experimental Rattus norvegicus model, focusing on the effects of benzodiazepine and alcohol ingestion. Twenty-one male rats were divided into three groups: control (K), benzodiazepine-treated (B), and alcohol-treated (A). After two weeks of substance administration, hypothermia was induced and multiple organ samples were analyzed. Histologically, renal tissue showed hydropic and vacuolar degeneration, congestion, and acute tubular injury across all groups, with no significant differences in E-cadherin expression. Lung samples revealed congestion, emphysema, and hemorrhage, with more pronounced vascular congestion in the alcohol and benzodiazepine groups. Cardiac tissue exhibited vacuolar degeneration and protein denaturation, particularly in substance-exposed animals. The spleen showed preserved architecture but increased erythrocyte infiltration and significantly elevated myeloperoxidase (MPO)-positive granulocytes in the intoxicated groups. Liver samples demonstrated congestion, focal necrosis, and subcapsular hemorrhage, especially in the alcohol group. Immunohistochemical analysis revealed statistically significant differences in MPO expression in both lung and spleen tissues, with the highest levels observed in the benzodiazepine group. Similarly, CK7 and CK20 expression in the gastroesophageal junction was significantly elevated in both alcohol- and benzodiazepine-treated animals compared to the controls. In contrast, E-cadherin expression in the kidney did not differ significantly among the groups. These findings suggest that specific histological and immunohistochemical patterns, particularly involving pulmonary, cardiac, hepatic, and splenic tissues, may help differentiate primary hypothermia from substance-related secondary hypothermia. The study underscores the value of integrating toxicological, histological, and molecular analyses to enhance the forensic assessment of hypothermia-related fatalities. Future research should aim to validate these markers in human autopsy series and explore additional molecular indicators to refine diagnostic accuracy in forensic pathology. Full article

Background: Proximal gastrectomy (PG) with double tract reconstruction (DTR) offers organ preservation for early gastric cancers, leading to reduced vitamin B12 deficiency, less weight loss, and improved quality of life. The JCOG1401 study confirmed excellent long-term outcomes for PG in stage I gastric cancer. However, in locally advanced proximal gastric cancer (LAPGC), preserving the gastric body and lymph node station 4d may compromise margin clearance and adequate lymphadenectomy. Methods: We propose a modified PG that removes the distal esophagus, gastroesophageal junction (GEJ), cardia, fundus, and gastric body, preserving only the antrum and performing DTR. Lymphadenectomy is also adapted, removing stations 1, 2, 3a, 4sa, 4sb, 4d, 7, 8, 9, 10 (spleen preserving), 11, and lower mediastinal nodes (stations 19, 20, and 110), while preserving stations 3b, 5, and 6. Indications for this procedure include GEJ (Siewert type II and III) and proximal gastric cancers with ≤2 cm distal esophageal involvement and ≤5 cm gastric involvement. Results: In our initial experience with 14 patients, we achieved R0 resection in all patients, adequate lymph node harvest (median 24 nodes, IQR 18–38), and no locoregional recurrences at a median follow-up of 18 months. We also found favorable postoperative weight loss, reflux, and anemia in the PG cohort. Conclusion: While larger studies and long-term data are still needed, our early results suggest that modified PG—despite sparing only the antrum—retains the key benefits of PG over total gastrectomy, including better weight maintenance and improved hemoglobin levels, while maintaining oncologic outcomes for LAPGC. Full article

Egg yolk immunoglobulin Y (IgY) possesses advantages such as low cost, easy availability, simple preparation, high antigen specificity, absence of drug residues, and compliance with animal welfare standards, making it an environmentally friendly and safe alternative to antibiotics. This research utilizes IgY antibody technology to develop a multivalent passive immune vaccine for major pathogenic bacteria in aquaculture. In this study, IgY antibodies against live Shewanella xiamenensis (LSX-IgY) and inactivated S. xiamenensis (ISX-IgY) were prepared by immunizing laying hens, and passive immunization protection experiments were conducted in Carassius auratus infected with S. xiamenensis and Aeromonas hydrophila. The passive immunization protection rates of LSX-IgY and ISX-IgY against S. xiamenensis were 63.64% and 72.73%, respectively, and the passive cross-protection rates against A. hydrophila were 50% and 71.43%, respectively. Further, C. auratus sera could specifically bind to S. xiamenensis or A. hydrophila in vitro, and the phagocytic activity of leukocytes was increased. LSX-IgY and ISX-IgY could reduce the bacterial load in the C. auratus kidneys. Meanwhile, they could significantly reduce the levels of antioxidant factors in serum and inhibit the mRNA expression of inflammation-related factors in the kidneys and spleens. Additionally, histopathology and immunofluorescence analysis showed that both IgY preparations preserved tissue integrity and reduced the expression of apoptosis factor (p53) and DNA damage factor (γH2A.X) of visceral organs, respectively. In summary, LSX-IgY and ISX-IgY can combat various bacterial infections, with no significant difference between the two. Additionally, inactivated bacterial immunization is more aligned with animal welfare standards for laying hens. Therefore, ISX-IgY is expected to serve as a multivalent vaccine against major aquaculture pathogens. Full article

Background/Objectives: Splenic torsion is a well-known and reported clinical problem. Splenic torsions after abdominal trauma represent a small group of cases that involve surgical management. They manifest primarily as abdominal pain, and the diagnosis is made based on imaging studies—ultrasound, CT, and MRI. Methods: This work aimed to analyze traumatic splenic torsions in terms of their clinical course, symptoms, timing, involvement of imaging techniques in the diagnosis, histopathological examination, and overall outcome. We searched databases using the desk research method under the keywords “splenic torsion”, “torsion”, and “spleen”, as well as in combination with “traumatic”, finding a total of eight cases, which we included in our analysis. Results: The eight cases were analyzed, comprising four females and four males, with an average age of 16.25 years (range 5–29 years). Traffic accidents were the most frequent cause of injury (five cases), while the circumstances were unclear in the remaining three. Immediate abdominal symptoms appeared in six patients. Splenic torsion was preoperatively diagnosed in five out of seven confirmed cases. A total of seven patients underwent laparotomy with splenectomy. In one case, laparoscopy converted to laparotomy with splenopexy preserved the spleen. Histopathology, performed in only two cases, confirmed splenic infarction in one patient; infarction status could not be determined in the remaining five due to missing data. Conclusions: Post-traumatic splenic torsions are a group of atypical injuries as the primary and immediate consequence of the trauma suffered is not anatomical–structural damage to the organ, such as a rupture. Mostly affecting young people, the cases described in the professional literature involve the main spleen, which was considered to be “wandering”, suggesting that this is a key predisposing factor for splenic torsion following blunt trauma and requiring diagnostic imaging for diagnosis. Full article

Background/Objectives: Challenges in normothermic machine perfusion (NMP) remain, particularly concerning the duration for which individual organs can be safely preserved. We hypothesize that optimal preservation can be achieved by perfusing organs together in a multivisceral block. Therefore, our aim was to establish a platform for ex situ multivisceral organ perfusion. Methods: Multivisceral grafts containing the liver, kidneys, pancreas, spleen, and intestine were obtained from Yorkshire pigs. Three generation (gen) set-ups were tested during the iterative design process, and minor changes were made throughout. Gen 1 (n = 4) used a custom-designed single perfusion circuit. Gen 2 (n = 3) employed a dual perfusion circuit. Gen 3 (n = 4) featured a single perfusion circuit with an optimized basin and reservoir. Grafts underwent NMP using an autologous blood-based perfusate, while hemostatic parameters and function were assessed. Results: Comparing Gen 1 versus Gen 3, the mean aortic flow improved (1.018 vs. 2.089 L), resistance decreased (0.224 vs. 0.038), urine output increased (51.90 vs. 271.3 mL), oxygen consumption rose (43.56 vs. 49.52 mL O₂/min), perfusate lactate levels dropped (10.44 vs. 3.10 mmol/L), and the pH became more physiological (7.27 vs. 7.30). Cellular injury trended lower in Gen 3. Histological evaluation demonstrated minimal differences in Gens 2 and 3. Conclusions: We demonstrate the feasibility of abdominal multivisceral NMP for up to 8 h. Adequate arterial flow, stable perfusate pH, and high oxygen consumption in setup 3 indicated organ viability. Multivisceral perfusion may serve as a plat-form for long-term NMP. Full article

Background/Objectives: The emergence of multidrug-resistant Acinetobacter baumannii (MDR A. baumannii) as a leading cause of fatal hospital-acquired infections underscores the urgent need for effective vaccines. While oral vaccines using live Bacillus subtilis spores expressing A. baumannii TonB-dependent receptor (TBDR) show promise, biosafety concerns regarding recombinant spore persistence necessitate alternative strategies. Here, we evaluated chemically inactivated B. subtilis spores displaying TBDR as a safer yet immunogenic vaccine candidate. Methods: Recombinant spores were inactivated using iron-ethanol sporicidal solution and administered to BALB/c mice (8–12 weeks old) to assess safety and immunogenicity. Toxicity was evaluated through clinical monitoring, serum biochemistry, and histopathology. Immune responses were characterized by T/B cell activation, IgG/IgA titers, and mucosal sIgA levels. Protective efficacy was determined by challenging immunized mice with MDR A. baumannii Ab35 and quantifying bacterial loads and examining tissue pathology. Results: The inactivated spores exhibited an excellent safety profile, with no adverse effects on clinical parameters, organ function, or tissue integrity. Immunization induced robust systemic and mucosal immunity, evidenced by elevated CD4+/CD8+ T cells, B cells, and antigen-specific IgG/IgA in serum and mucosal secretions. Following the challenge, vaccinated mice showed significantly reduced pulmonary bacterial burdens (>90% reduction), and preserved lung and spleen architecture compared to controls, which developed severe inflammation and tissue damage. Conclusions: These findings demonstrate that inactivated B. subtilis spores expressing TBDR are a safe, orally administrable vaccine platform that elicits protective immunity against MDR A. baumannii. By addressing biosafety concerns associated with live spores while maintaining efficacy, this approach represents a critical advance toward preventing high-risk nosocomial infections. Full article