Histological Assessment Research Articles

Application of stem cells alone and in combination with low-level laser therapy for sciatic nerve repair in rats

This study evaluated the efficacy of tubular constructs containing stem cells and Type I collagen, both independently and in conjunction with low-level laser therapy (LLLT), in repairing the sciatic nerve in a rat model. In this animal study, the right sciatic nerve of 30 male Wistar rats, each weighing 250–300 g, was surgically excised to a length of 8 mm. The rats were then randomly allocated to three groups (n = 10 per group). In Group 1, the excised nerve segment was utilized as an autograft and sutured at the defect site. In Group 2, a tubular construct containing stem cells and Type I collagen was used to bridge the proximal and distal ends of the nerve. Group 3 received the same intervention as group 2, supplemented with 5 weeks of LLLT. After 5 and 12 weeks, rats underwent histological, behavioral, and electrophysiological assessments. Data were statistically analyzed using analysis of variance (ANOVA), Bonferroni post-hoc test, and Kruskal–Wallis test. At both 5 and 12 weeks, axonal count and nerve repair scores showed no significant differences among the three groups (P > 0.05). Notably, the Sciatic Functional Index (SFI) was the most favorable (lowest) in the autograft group, whereas the stem cell-only group exhibited the least favorable (highest) SFI at 5 weeks (P < 0.001). Additionally, distal latency was highest in the stem cell group and lowest in the stem cell combined with LLLT group at 5 weeks (P < 0.001). A significant difference was observed between the autograft and stem cell plus LLLT groups (P < 0.05). In conclusion, the application of stem cell-laden tubular constructs in conjunction with LLLT demonstrated efficacy for sciatic nerve repair in rats.

Serology and histology in Sjögren's syndrome diagnosis: a retrospective accuracy study.

Sjögren's syndrome (SS) presents complex diagnostic challenges due to its multi-organ involvement, often leading to misdiagnosis, which can result in unnecessary treatments, elevated healthcare costs, and significant impacts on patient quality of life. Accurate diagnosis is therefore critical, utilising ACR/EULAR criteria that include both labial minor salivary gland (LMSG) biopsy and anti-SSA antibodies. This retrospective study analysed medical records of 87 adults suspected of primary SS, who underwent both anti-SSA serology and LMSG biopsy. We evaluated the diagnostic accuracy of these tests under existing ACR/EULAR criteria and a newly proposed 'modified ACR/EULAR criteria-ClinDx'. Statistical analysis included Pearson's chi-square test for the association between test results and disease status and receiver operating characteristic (ROC) curves to assess the sensitivity and specificity of the diagnostic models. Utilising ACR/EULAR criteria, 40 patients were diagnosed with SS, while 47 were categorised as non-diseased. The ClinDx criteria application resulted in 32 SS diagnoses and 55 non-diseased classifications, highlighting discrepancies in patients with low anti-SSA titers (< 200 MFU). Statistical analysis confirmed a significant association (p < 0.001) between test results and disease status, indicating the robustness of the modified criteria in enhancing diagnostic accuracy. This study underscores the utility of integrating serological tests and histological biopsies in diagnosing SS. While anti-SSA antibodies provide a good preliminary screening tool, the specificity of LMSG biopsies is indispensable. Refining both serological and histological assessments per ClinDx criteria can improve diagnostic accuracy, aiding in better management of SS and reducing healthcare burdens.

Comparative Histological Assessment of Zinc Oxide Nanoparticles and Low-Power Laser Treatment at 810nm Wavelength on the Recovery of Second-Degree Burn Wounds in Rat Models.

Background: The utilization of zinc oxide nanoparticles is thought to augment wound healing because of their antibacterial characteristics and capacity to stimulate cellular regeneration, especially in instances of minor burn injuries. On the other hand, it has been shown that tissue regeneration is aided by low-power laser therapy via photobiomodulation. Zinc oxide nanoparticles and low-power laser therapy are the two therapeutic modalities that will be compared in this study in order to assess how well they promote healing after burn injury and provide important new information on improved wound care techniques. Methods: For this investigation, thirty male Wistar rats weighing 230 ± 25 grams each were split into three groups. Every rat received general anesthesia before the experiment. A stainless-steel rod was put to the rats' skin after being heated for 20 min in a boiling water bath to cause superficial second-degree burns. The control group functioned as a reference point for comparison and did not receive any treatment intervention. Over the course of a week, zinc oxide nanoparticles were applied topically to the second group. For one week, the third group received daily therapy with a diode laser at a dosage of 10 J/cm.1 Histological and clinical exams were performed after the therapy period to evaluate the impact of the therapies. Results: The experimental groups that received low-power laser therapy (third group) and zinc oxide nanoparticles (second group) showed a substantial increase in wound contraction in relation to the control group, based on macroscopic observations. One rat from the second group showed notable indications of full wound healing on day 21. The treated rats showed the highest rate of lesion contraction, indicating that wound treatment happened at least 7 days faster in these rats than in the other groups. After 21 days, the third group's epidermis fully epithelized and formed a layer of keratinization. Furthermore, there was enhanced angiogenesis and significant fibroblast proliferation; large-scale fibrosis was also commonly seen. Conclusion: Zinc oxide nanoparticles promoted wound healing and accelerated connective tissue regeneration faster than other groups when applied to second-degree superficial burns. This research implies that the use of zinc oxide nanoparticles may be a therapeutic approach that shows promise for treating burn injuries and improving patient outcomes.

Exploring Azithromycin’s Neuroprotective Role in Traumatic Brain Injury: Insights into Cognitive and Motor Recovery and Neuroinflammatory Modulation

Background: Traumatic brain injury (TBI) is a leading cause of mortality worldwide and often results in substantial cognitive, motor, and psychological impairments, triggering oxidative stress, neuroinflammation, and neurodegeneration. This study examined the neuroprotective effects of azithromycin (AZI) in TBI. Methods: TBI was induced in rats using the weight-drop method. Subsequently, rats received a daily intraperitoneal (I.P.) dose of AZI (150 mg/kg) for 28 days. Behavioral tests (Morris water maze, rotarod, and open field tests) were performed to assess cognitive and motor functions. Neurochemical analyses included oxidative stress markers (GSH, SOD, MDA, catalase), inflammatory cytokines (TNF-α, IL-1β), apoptotic markers (caspase-3, Bax, Bcl-2), mitochondrial complexes (complex I, II, III, IV, and V), and the transforming growth factor- beta (TGF-β) as a neurofilament marker. Histological evaluations focused on neuronal integrity in the cortex, hippocampus, and striatum. Results: Treatment with AZI significantly facilitated motor and cognitive function recovery in TBI-affected rats. At the molecular level, AZI effectively reduced oxidative stress markers, ameliorated neuroinflammation by decreasing TNF-α, IL-1β, and neuronal apoptosis, and differentially modulated mitochondrial complexes. Histological assessments revealed enhanced neuronal integrity and fewer pathological changes in AZI-treated rats compared to untreated TBI controls. Conclusion: AZI was shown to interfere with several pathways involved in TBI’s pathophysiology. While preclinical results are promising, further studies are necessary to establish the long-term safety and efficacy of AZI in a clinical setting. This research supports the potential re-purposing of AZI as a novel treatment strategy for TBI and related neurodegenerative disorders.

Parameter-Tuned Pulsed Wave Photobiomodulation Enhances Stem Cells From Apical Papilla Differentiation: Evidence From Gene and Protein Analyses.

This study examines the effects of pulsed wave photobiomodulation (pwPBM) on the osteogenic differentiation of stem cells from the apical papilla (SCAP). Using 810 nm near-infrared (NIR) light with 300 Hz pulses and a 30% duty cycle, pwPBM was applied at a total energy density of 750 mJ/cm2. Osteogenesis was evaluated through both invitro and invivo analyses. Invitro experiments demonstrated significant enhancement of alkaline phosphatase (ALP) activity, along with upregulation of key osteogenesis-related genes and proteins, as confirmed by real-time polymerase chain reaction (PCR) and Western blot analyses. Invivo, histological assessments following SCAP transplantation revealed increased bone tissue formation, further corroborated by osteocalcin staining. These findings underscore the potential of pwPBM as an innovative and effective tool for dental tissue regeneration and engineering.

A Bioabsorbable Implant Seeded with Adipose-Derived Stem Cells for Adipose Regeneration.

Adipose tissue engineering requires effective strategies for regenerating adipose tissue, with adipose-derived stem cells (ASCs) being favored due to their robust self-renewal capacity and multipotent differentiation potential. In this study, the efficacy of poly-L-lactic acid (PLLA) mesh containing collagen sponge (CS), seeded with ASCs to promote adipose tissue formation, was investigated. PLLA-CS implants seeded with GFP-positive ASCs were inserted at high concentration (1 × 106 cells/implant, H-ASC) and low concentration (1 × 105 cells/implant, L-ASC), as were unseeded controls. Adipogenesis was evaluated at 3, 6, and 12 months using a rat inguinal model. At 3 months, the weight and volume of newly formed tissues in the H-ASC group were significantly higher than those in the control group. Histological assessment revealed that the area of all newly formed tissue, including the adipose tissue inside the implants in the H-ASC group, was larger at 6 and 12 months compared with that of the control and L-ASC groups, with the adipose percentage at 12 months being higher in the H-ASC group than in the control group. GFP-positive ASCs in both the L-ASC and H-ASC groups adhered to the CS scaffolds and survived for up to 12 months postimplantation, with spontaneous differentiation into adipocytes observed exclusively in the H-ASC group. Double immunofluorescence confirmed the presence of GFP-positive adipocytes. In summary, this study demonstrated that ASCs coimplanted with PLLA-CS implants could enhance adipose tissue formation within the implants. Uninduced ASCs were capable of spontaneously differentiating into adipocytes within the PLLA-CS implants, with differentiation correlating with the number of implanted cells.

Histology Assessment of Chitosan–Polyvinyl Alcohol Scaffolds Incorporated with CaO Nanoparticles

Scaffolds for regenerative therapy can be made from natural or synthetic polymers, each offering distinct benefits. Natural biopolymers like chitosan (CS) are biocompatible and biodegradable, supporting cell interactions, but lack mechanical strength. Synthetic polymers like polyvinyl alcohol (PVA) provide superior mechanical strength and cost efficiency but are not biodegradable or supportive of cell adhesion. Combining these polymers optimizes their advantages while adding metal oxide nanoparticles like calcium oxide (CaO NPs) enhances antimicrobial properties by damaging bacterial membranes. In this study, we obtained the formation of CaO NPs by calcinating eggshells, which were mixed in a polymeric network of CS and PVA to obtain four different membrane formulations for subdermal tissue regeneration. The spherical nanoparticles measured 13.43 ± 0.46 nm in size. Their incorporation into the membranes broadened the hydroxyl bands in the Fourier transform infrared (FTIR) analysis at 3331 cm⁻1. X-ray diffraction (XRD) analysis showed changes in the crystalline structure, with new diffraction peaks at 2θ values of 7.2° for formulations F2, F3, and F4, likely due to the increased amorphous nature and concentration of CaO NPs. Additionally, higher CaO NPs concentrations led to a reduction in thermal properties and crystallinity. Scanning electron microscopy (SEM) revealed a heterogeneous morphology with needle-like structures on the surface, resulting from the uniform dispersion of CaO NPs among the polymer chains and the solvent evaporation process. A histological examination of the implanted membranes after 60 days indicated their biocompatibility and biodegradability, facilitated by incorporating CaO NPs. During the degradation process, the material fragmented and was absorbed by inflammatory cells, which promoted the proliferation of collagen fibers and blood vessels. These findings highlight the potential of incorporating CaO NPs in soft tissue regeneration scaffolds.

The association between skin allergy testing and oral squamous cell carcinoma in oral lichen planus: a retrospective cohort study.

There is a reported association between oral contact allergy and oral lichen planus (OLP). Likewise oral squamous cell carcinoma (oSCC) is associated with OLP. It is hypothesized that chronic inflammation may contribute to oSCC risk. Ostensibly, we hypothesized that allergy testing positivity may increase inflammation and thus may be associated with oSCC in OLP. As a secondary objective, we assessed oSCC prevalence in OLP, allergen prevalence in OLP, and associations between allergy testing and OLP phenotype. To do this, we performed a retrospective cohort evaluation of OLP patients seen at the University of Utah from 2015 to 2022. Odds of oSCC occurrence by allergy testing status, clinical/demographic factors and asssociations between allergy testing and OLP phenotype were assessed using univariable and multivariable logistic regression. The prevalence of oSCC and allergy testing (patch + scratch testing) results were summarized descriptively. OLP diagnoses were confirmed by our OLP-specialized clinicians, and/or histologic assessment. Allergy testing was performed by two specialized dermatologists. oSCC diagnosis was identified via chart review and histologic data. Among 587 OLP patients identified, 133 were allergy tested and 77.4% were positive (52.4% metals, 55.3% flavorings/fragrances, 47.6% preservatives). Of those with a positive allergy test, 10 (25.6%) developed oSCC compared with 2 (5.1%) of patients with a negative allergy test. There was insufficient evidence to support an association between allergy positivity and oSCC (OR = 1.55, 95% CI 0.76-3.15). A higher percent of symptomatic visits was the only other clinical and/or demographic factor associated with oSCC occurrence (median 87.5% vs. 66.7%, p = 0.03). Of the OLP disease characteristics assessed (mouth area involved, extent of disease involvement, erosive subtype, etc.), only OLP involving the gingiva was associated with positive allergy testing (OR = 2.54,1.11-5.81). These data suggest that allergy test positivity may not be associated with oSCC. However, it was associated with more pervasive symptomatology, suggesting a possible association with more severe or recalcitrant disease.

Defining an NK Cell-enriched Rejection-like Phenotype in Liver Transplant Biopsies From the INTERLIVER Study.

Initial analysis of liver transplant biopsies in the INTERLIVER study (ClinicalTrials.gov; unique identifier NCT03193151) using rejection-associated transcripts failed to find an antibody-mediated rejection state (ie, rich in natural killer [NK] cells and with interferon-gamma effects). We recently developed an optimization strategy in lung transplants that isolated an NK cell-enriched rejection-like (NKRL) state that was molecularly distinct from T cell-mediated rejection (TCMR). Here we apply the same strategy to a liver transplant biopsy population. We used this strategy to search for a molecular NKRL state in 765 consented liver transplant biopsies collected at participating international centers for gold-standard histology and molecular assessment by genome-wide microarrays. Validation through a training set-test set approach of an optimized selection of variables as inputs into unsupervised rejection classification identified an NKRL state in livers. The full model classified 765 biopsies into the following molecular phenotypes, characterized by their gene expression: no-rejection 54%, TCMR 16%, NKRL 13%, and injury 16%. Top TCMR transcripts were expressed in effector T cells; top NKRL transcripts were almost exclusively expressed in NK cells; and both had increased interferon-γ-inducible transcripts, which were more pronounced in TCMR. Most TCMR biopsies had significant parenchymal injury, molecular fibrosis, and abnormal biochemistry. NKRL biopsies had no excess of injury, fibrosis, or biochemistry abnormalities. Optimized rejection algorithms indicate that some liver transplants manifest an NKRL state that is well tolerated in the short term postbiopsy and with minimal injury and relatively normal biochemistry, while also underscoring the potential of TCMR to produce extensive parenchymal injury.

Efficacy and safety of resmetirom in MASLD and MASH: network meta-analysis of randomized clinical trials.

Metabolic-Associated Steatohepatitis-Related Liver Disease (MASLD) and, its progressive form, Metabolic-Associated Steatohepatitis (MASH) pose significant global health challenges. Current therapeutic strategies targeting metabolic abnormalities have shown promise but lack specificity for the liver. Thyroid hormones, particularly thyroid hormone receptor beta (THR-β) agonists like resmetirom, offer a targeted approach to liver-related pathways. A network meta-analysis (NMA) comparing different doses of resmetirom to placebo for MASLD and MASH was conducted. PubMed, Scopus, Cochrane, and Webof Science were searched for relevant randomized controlled trials (RCTs). Efficacy outcomes included histological, radiological, and biochemical parameters, while safety outcomes comprised adverse events and treatment discontinuation. Resmetirom demonstrated dose-dependent efficacy in histological and radiological assessments, with the 100 mg dose showing superior MASH resolution and hepatic fat reduction. Biochemical markers indicated improved liver function with resmetirom treatment. However, adverse events, particularly diarrhea and nausea, were more prevalent in the resmetirom group, leading to higher treatment discontinuation rates. Resmetirom shows promise as a therapeutic option for MASLD and MASH management, with significant improvements in liver health parameters. However, safety concerns warrant careful monitoring in clinical practice. Further research is needed to optimize its long-term safety and efficacy.

SFAs facilitates ceramide's de novo synthesis via TLR4 and intensifies hepatocyte lipotoxicity.

Non-alcoholic steatohepatitis (NASH), an advanced manifestation of non-alcoholic fatty liver disease (NAFLD), is characterized by hepatocyte injury, inflammation, and fibrosis. Saturated fatty acids (SFAs) have emerged as key contributors to hepatocyte lipotoxicity and disease progression. Toll-like receptor 4 (TLR4) acts as a sentinel for diverse ligands, including lipopolysaccharide (LPS) and endogenous molecules like palmitic acid (PA)-induced ceramide (CER) accumulation, promoting hepatocyte demise. However, the intricate mechanisms underlying TLR4's modulation of ceramide metabolism and their concerted effect on SFA-mediated hepatotoxicity remain elusive. A NASH mouse model with liver-specific TLR4 knockdown was established through palm oil feeding and AAV2/8 tail vein injection. Histological and biochemical assessments were conducted to evaluate the mice's condition and liver damage extent. Liquid chromatography-mass spectrometry (LC-MS) was employed to quantify ceramide levels in liver tissues, offering insights into NASH mechanisms. The PO-fed model exhibited elevated serum ALT, AST, and liver TG levels, enhancing lipid accumulation and hepatocellular damage. TLR4 knock-down reduced liver mass and the liver-to-body weight ratio, signifying a decreased hepatic burden. Histopathological evaluations revealed substantial improvement in hepatic steatosis in TLR4-silenced PO-fed mice, with diminished lipid droplets and inflammatory infiltrates. LC-MS analysis showed a marked decrease in long-chain ceramides (C14, C16, C20) in TLR4-knockdown PO-fed mice. Furthermore, expression of MyD88, SPTLC1, SPTLC2, and inflammatory markers IL-1β, IL-6, TNF-α were significantly attenuated. SFAs activate the TLR4 signaling pathway via MyD88, fostering ceramide de novo synthesis, which exacerbates hepatocyte lipotoxicity and accelerates NASH progression.

Antimicrobial Potentials of Haliclona fibulata Derived Compounds for Combatting Drug-Resistant Bacterial Infections in Zebrafish Model.

The rising threat of antimicrobial resistance among pathogens highlights the critical need for novel antimicrobial agents. This study explores the potential of natural products by investigating hexane extracts from the marine sponge Haliclona fibulata (HF) for their antibacterial efficacy. The well diffusion method of HF extract showed significant antibacterial activity against P. aeruginosa, E. coli and S. aureus with inhibition zones of 20 ± 0.66 mm, 27 ± 0.58 mm, and 22 ± 0.33 mm, respectively. Scanning electron microscopy investigations confirmed the disruptive impact of the extracts on bacterial cell morphology. Further investigations using P. aeruginosa-infected zebrafish model revealed that HF treatment improved survival rates in a dose-dependent manner. Additionally, oxidative stress and apoptosis were significantly reduced in the 100 μg/mL (HF100) treatment group, alongside down-regulation of inflammatory markers such as il-1β, nf-kβ, tnf-α, and inos. Histological assessments showed reduced intestinal damage and fewer circulating macrophages in HF100-treated zebrafish larvae. Identifying potent bioactive compounds within the extracts suggests that HFcould be a valuable source of natural antimicrobials, warranting further exploration for developing treatments against pathogenic infections.

Protaetia brevitarsis Hydrolysate Mitigates Muscle Dysfunction and Ectopic Fat Deposition Triggered by a High-Fat Diet in Mice

Background/Objectives: Obesity is a key factor in metabolic syndrome (MetS) development. Consumption of a high-fat diet (HFD) accelerates the onset of obesity and associated metabolic complications. Protaetia brevitarsis (PB) has been traditionally utilized in Korean medicine for its antioxidant, anti-diabetic, anticancer, and hepatoprotective effects. However, specific effects of PB hydrolysate on skeletal muscles have not been fully elucidated. Therefore, this study sought to assess the influence of PB on HFD-induced MetS, focusing on the lipid metabolism and inflammatory responses mediated by AMP-activated protein kinase activation. Methods: To induce obesity, 6-week-old C57BL/6J mice were maintained on an HFD for 8 weeks, after which PB hydrolysate was orally administered for 16 weeks while the HFD regimen was sustained. A glucose tolerance test was conducted orally to evaluate glucose regulation, and forelimb grip strength was assessed upon completion of the experimental period. Histological assessments, serum biochemical analysis, lipid extraction, Western blot analysis, and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) were performed following euthanasia. Results: PB significantly reduced ectopic lipid deposition in skeletal muscles, enhanced muscle strength, and improved insulin sensitivity by increasing fatty acid oxidation via AMP-activated protein kinase/carnitine palmitoyltransferase 1 activation and inhibiting lipogenesis via stearoyl-CoA desaturase 1 gene downregulation. Furthermore, PB alleviated HFD-induced low-grade chronic inflammation by decreasing systemic monocyte chemoattractant protein 1 levels, thereby reducing ectopic fat deposition. Conclusions: This study highlights the potential of PB as a nutraceutical to mitigate MetS in HFD-fed mice.

Comparison of cellular-based therapies following a long-segmental peripheral nerve defect in a rat model.

Peripheral nerve injury (PNI) is characterized by a loss of cellular and axonal integrity, often leading to limited functional recovery and pain. Many PNIs are not amenable to repair with traditional techniques; however, cell therapies, particularly Schwann cells (SCs), offer the promise of neural tissue replacement and functional improvement. Exosomes, which carry cellular signaling molecules, can be secreted by SCs and have shown promise in PNI. Our laboratory has had success using SCs in preclinical and clinical treatment settings. Transplanted cells have several known limitations, which exosomes mitigate. To that end, the current study investigated if implanted SC-derived exosomes in conduits, conduits with SCs, reverse autograft, or empty conduits comparably improve axonal regeneration and pain outcomes 16-weeks after repair of a long gap PNI in adult rats. Results show that there were no differences between groups in the von Frey filament testing or in the Hargreaves test. Electrophysiological testing showed a significant difference between the injured (ipsilateral) and uninjured (contralateral) limbs while histological assessment showed a significant difference between axonal counts in different areas of the conduit. Based on the results of the current study, more research is needed to understand the therapeutic role of exosomes in PNI.

A Quantitative Method for Assessing Treatment-Related Changes within the Airway Mucosa in Patients with Chronic Bronchitis

No standardized method has yet been established for evaluating airway mucosal aberrancies associated with chronic obstructive pulmonary disease (COPD) or chronic bronchitis (CB). While goblet cell hyperplasia (GCH) is an established pathognomonic hallmark of the CB disease process, no standardized method exists for acquiring mucosal biopsies and assessing morphologic airway mucosa alterations. Additionally, the impacts from interventions targeting the airway mucosa are not well defined. In this context, a reliable and robust measure for assessing airway mucosa at baseline and subsequent to an intervention is critical for characterizing treatment-related changes. Can standardizing airway biopsy tissue collection and histopathological assessment methods generate a robust and repeatable measure to assess airway mucosa tissue characteristics in the setting of COPD/CB? Initial tissue collection and histological assessment methods were designed by integrating various aspects from previously published evaluations, applied to an initial tissue sample cohort, and then iteratively refined by independent pathologists. A standardized metric for histologic airway mucosa assessments was developed that specified tissue collection methods, including re-sampling airways at multiple time points to enable evaluation of treatment-related effects by incorporating scores for GCH, eosinophilia, and chronic inflammation, and the degree of GCH heterogeneity present within each sample. This multi-center study generated a robust, reproducible approach for assessing airway mucosa aberrancies in the setting of COPD/CB. The iterative approach established consistent tissue specimen recovery and a granular scoring matrix that enabled quantitative scoring of the various tissue findings with substantial histopathologic interrater reliability. ClinicalTrials.gov; NCT03107494, NCT04677465; URL: www. gov. Australian New Zealand Clinical Trials Registry (ANZCTR); ACTRN12617000330347; URL: anzctr.org.au.

Heat acclimation mediates cellular protection via HSP70 stabilization of HIF-1α protein in extreme environments.

Heat acclimation (HA) is an evolutionarily conserved trait that enhances tolerance to novel stressors by inducing heat shock proteins (HSPs). However, the molecular mechanisms underlying this phenomenon remain elusive. In this study, we established a HA mouse model through intermittent heat stimulation. Subsequently, this model was evaluated using an array of physiological and histological assessments. In vitro, HA cell model with mouse brain microvascular endothelial cells (bEnd.3) was established and analyzed for cell viability and apoptosis markers. We investigated HA-mediated heat and hypoxia tolerance mechanisms using HIF-1α and HSP70 inhibitors and siRNA. Our results demonstrated that HA enhances the tolerance of bEnd.3 cells and mice to both heat and hypoxia, Mechanistically, HA upregulated the expression of HIF-1α and HSP70. However, inhibition of HIF-1α or HSP70 partially attenuated HA-induced tolerance to heat and hypoxia. Additionally, HA significantly decreased the ubiquitination levels of HIF-1α, whereas inhibition of HSP70 increased its ubiquitination. HA also substantially enhanced the interaction between HIF-1α and HSP70. In conclusion, our findings indicate that HA enhances tolerance to heat and hypoxia by stabilizing HIF-1α through increased interaction with HSP70. This discovery elucidates a novel mechanism of cellular protection conferred by HA and provides new strategies and potential targets for human adaptation to extreme environments.

Canagliflozin attenuates neurodegeneration and ameliorates dyskinesia through targeting the NLRP3/Nurr1/GSK-3β/SIRT3 pathway and autophagy modulation in rotenone-lesioned rats.

This study investigated for the first time the neuroprotective effect of canagliflozin against PD and LID in rotenone-intoxicated rats, emphasizing the crosstalk among the NLRP3/caspase-1 cascade, PGC-1α/SIRT3 pathway, mammalian target of rapamycin (mTOR)/beclin-1, and Nurr1/β-catenin/GSK-3β pathways as possible treatment strategies in PD and LID. Also, correlating NLRP3 expression with all evaluated parameters. The PD rat model was induced via eleven rotenone (1.5mg/kg) subcutaneous injections day after day. Canagliflozin (20mg/kg) and/or L-dopa/carbidopa (100/25mg/kg) were orally administered daily from the beginning until the end of the experiment. Canagliflozin significantly improved neurobehavioral and histological assessments, whereas dyskinesia scores declined. The improvement was confirmed through tyrosine hydroxylase and β-catenin upregulation in contrast to NLRP3 and caspase-1 in substantia nigra pars compacta, as revealed immunohistochemically. In addition, canagliflozin induced a prominent elevation in dopamine, Nurr1, PGC-1α, SIRT3, and beclin-1, whereas mTOR and GSK-3β expressions were downregulated. Our results revealed the aspiring canagliflozin neuroprotective properties against PD and LID in rotenone-lesioned rats via the assumed anti-inflammatory activity and implication of NLRP3/caspase-1, Nurr1/GSK-3β/β-catenin, PGC-1α/SIRT3, and beclin-1/mTOR pathways.

Multi-omics profiling reveals altered mitochondrial metabolism in adipose tissue from patients with metabolic dysfunction-associated steatohepatitis.

Metabolic dysfunction-associated steatotic liver disease (MASLD) and its more severe form steatohepatitis (MASH) contribute to rising morbidity and mortality rates. The storage of fat in humans is closely associated with these diseases' progression. Thus, adipose tissue metabolic homeostasis could be key in both the onset and progression of MASH. We conducted a case-control observational research using a systems biology-based approach to analyse liver, abdominal subcutaneous adipose tissue (SAT), omental visceral adipose tissue (VAT), and blood of n=100 patients undergoing bariatric surgery (NCT05554224). MASH was diagnosed through histologic assessment. Whole-slide image analysis, lipidomics, proteomics, and transcriptomics were performed on tissue samples. Lipidomics and proteomics profiles were determined on plasma samples. Liver transcriptomics, proteomics, and lipidomics revealed interconnected pathways associated with inflammation, mitochondrial dysfunction, and lipotoxicity in MASH. Paired adipose tissue biopsies had larger adipocyte areas in both fat depots in MASH. Enrichment analyses of proteomics and lipidomics data confirmed the association of liver lesions with mitochondrial dysfunction in VAT. Plasma lipidomics identified candidates with high diagnostic accuracy (AUC=0.919, 95% CI 0.840-0.979) for screening MASH. Mitochondrial dysfunction is also present in VAT in patients with obesity-associated MASH. This may cause a disruption in the metabolic equilibrium of lipid processing and storage, which impacts the liver and accelerates detrimental adaptative responses. The project leading to these results has received funding from 'la Caixa' Foundation (HR21-00430), and from the Instituto de Salud Carlos III (ISCIII) (PI21/00510) and co-funded by the European Union.

Bacillus coagulans alleviates hepatic injury caused by Klebsiella pneumoniae in rabbits.

As an opportunistic bacterial pathogen, Klebsiella pneumoniae (KP) is prone to causing a spectrum of diseases in rabbits when their immune system is compromised, which poses a threat to rabbit breeding industry. Bacillus coagulans (BC), recognized as an effective probiotic, confers a variety of benefits including anti-inflammatory and antioxidant properties. The purpose of this study was to investigate whether dietary BC can effectively alleviate hepatic injury caused by KP. In this study, the rabbits were initially pretreated with varying doses of BC (1×106, 5×106, and 1×107 CFU/g), followed by a challenge with KP at a concentration of 1011 CFU/mL. Liver tissues were harvested and processed for histological assessment using H&E and VG stains to assess structural alterations. Biochemical assays were employed to quantify the enzymatic activities of T-SOD and GSH-Px, as well as the MDA content. Furthermore, ELISA was utilized to detect the levels of inflammatory cytokine (IL-10, IL-6, IL-1β and TNF-α) and apoptotic-related gene (Bcl-2, Bax). Morphological observation indicated that BC can effectively mitigate KP-induced hepatic sinusoidal dilatation and congestion, as well as ameliorate the degree of hepatic fibrosis. Further analysis showed that BC significantly lowered MDA level in KP-treated rabbits, while enhanced the activities of T-SOD and GSH-Px. Additionally, ELISA result showed that BC pretreatment significantly reduced the levels of pro-inflammatory cytokines TNF-a, IL-6, IL-1β and pro-apoptotic gene Bax, while increasing the levels of anti-inflammatory cytokine IL-10 and anti-apoptotic gene Bcl-2 in KP-treated rabbits. Above data indicate that BC supplementation effectively attenuated oxidative stress and inflammatory response induced by KP through augmenting the activities of antioxidant enzymes and diminishing the levels of pro-inflammatory factors. Furthermore, it reduced the Bax/Bcl-2 ratio in the liver, thereby inhibiting KP-induced apoptosis. The treatment group receiving 5x106 CFU/g BC benefitted most from the protective effect.

Usefulness of serial in vivo imaging to directly assess the role of inflammation in thrombus resolution and organization.

Deep vein thrombosis (DVT) remains a significant health problem. Although animal models have provided significant insights into the DVT pathophysiology, time-course assessment in a same animal is technically limited. Recently, we reported a novel murine saphenous DVT model for in vivo visualization of spatiotemporal dynamics of inflammatory cells. This study further shed a light on the resolution and organization process of DVT using serial in vivo imaging technique. Similar with ferric chloride-induced thrombus model, our saphenous DVT model allowed serial in vivo imaging with fluorescence microscopy. However, unlike ferric chloride-induced thrombus model, we observed a significant decrease of DVT burden. Red blood cells area gradually decreased followed by fibrin and collagen deposition over time, although ferric chloride model induced platelet-rich arterial thrombus. Histological assessment revealed that neutrophils influx peaked 3h after DVT induction, followed by macrophages' migration at 120h' post-induction, indicating similar organization process with traditional stasis-induced DVT model. Ly6G/Ly6C positive cells at 3h predicted the reduction of DVT burden (r>0.8; P<0.01), suggesting that inflammatory response at acute phase plays pivotal role in DVT resolution. MMP-9 expression was observed and colocalized with neutrophils at early timepoints in both traditional stasis-induced DVT model and our femoral imaging models. Taken together, our in vivo imaging model might allow better understanding of the resolution and organization processes in DVT.