Vehicle Group Research Articles

Fecal microbiota transplantation alleviates neuronal Apoptosis, necroptosis and reactive microglia activation after ischemic stroke

ObjectiveThis study aims to delve into the mechanisms underlying the improvement of neurological function in rats with ischemic stroke through fecal microbiota transplantation. MethodsA total of fifty male Sprague-Dawley rats were categorized into four groups: Sham, MCAO, MCAO+vehicle and FMT. We assessed behavioral and pathological alterations in the rats using modified neurological function scoring and TTC staining.Additionally, Western blot and immunofluorescence were used to detect the expression levels of Apoptotic and Necroptosis markers in neurons of ischemic brain tissue, and immunofluorescence was used to analyze the degree of activation of microglia. ResultsFMT group exhibited a decline in neurological function score compared to the MCAO and MCAO + vehicle group, accompanied by a reduction in infarct volume (P < 0.05). Relative to the SHAM group, the MCAO group displayed a significant increase in the expression levels of necroptosis-related proteins Phospho-RIP1, Phospho-RIP3, Phospho-MLKL, apoptotic proteins Bax and Cleaved caspase-3, and the iNOS positive microglia in ischemic brain tissue, while Bcl-2 expression was notably decreased (P < 0.05).Conversely, compared to the MCAO + vehicle group, the FMT group showed decreased expression levels of Phospho-RIP1, Phospho-RIP3, Phospho-MLKL, Bax, Cleaved caspase-3, and iNOS-positive microglia, while the expression of Bcl-2 was increased. ConclusionFecal microbiota transplantation offers a promising approach to improving neurological function in rats with ischemic stroke by inhibiting neuronal apoptosis, necroptosis, and the polarization of inflammatory microglial cells.

The Role of Probiotics in Modulating Myocardial Infarction and Depression-like Symptoms: A Study on Sex-Specific Responses.

This study explores the effects of two probiotics, Lactobacillus helveticus R0052 and Bifidobacterium longum R0175, on myocardial infarction (MI) and associated depression-like behaviours, with a focus on sex differences. MI was induced in adult male and female rats by occluding the left anterior coronary artery for 30 min, followed by 24 h of reperfusion. Probiotics were administered via drinking water for at least two weeks before ischemia. Infarct size, plasma C-reactive protein (CRP), estradiol levels, and intestinal permeability were then measured. Two weeks after, MI subgroups of rats were tested for depression-like behaviours. We found a significant interaction between sex and probiotics in relation to infarct size. Probiotics significantly reduced the infarct size compared to the vehicle group in female rats but not in males. Probiotics increased the plasma estradiol levels and reduced the CRP concentrations in females, suggesting anti-inflammatory and cardioprotective properties. Probiotics significantly increased intestinal resistance following MI in males only, suggesting sex-specific physiological responses to treatment. Probiotics enhanced social interaction in males with MI but not in females. Similarly, in the forced swim test, probiotics reduced immobility in males with MI but increased it in females, further underscoring the sex-dependent effects of probiotics. This study reports cardioprotective effects of probiotics upon MI in female rats, while benefits in male rats were rather at the behavioural level. These results highlight distinct physiological and behavioural responses between sexes, emphasizing the need to account for sex differences in future tests of probiotics as a prophylactic treatment for MI.

Tapinarof cream for the treatment of atopic dermatitis: Efficacy and safety results from two Japanese phase 3 trials.

Tapinarof is a nonsteroidal, topical, aryl hydrocarbon receptor agonist. We evaluated the efficacy and safety of tapinarof cream 1% in Japanese patients aged ≥12 years with atopic dermatitis (AD) in two phase 3 trials, ZBB4-1 and ZBB4-2. ZBB4-1 (N = 216) consisted of an 8-week, double-blind, vehicle-controlled treatment period (period 1) and a 16-week extension treatment period (period 2). Patients were randomized 2:1 to tapinarof or vehicle in period 1; subsequently, all patients who enrolled in period 2 received tapinarof. ZBB4-2 (N = 291) was a 52-week, open-label, uncontrolled trial in which all patients received tapinarof. In period 1 of ZBB4-1, the proportion of patients who achieved an Investigator's Global Assessment (IGA) score of 0 (clear) or 1 (almost clear) with ≥2-grade improvement from baseline at week 8 (IGA treatment success, the primary end point) was 20.24% in the tapinarof group and 2.24% in the vehicle group (p = 0.0007). The proportion of patients with ≥75% improvement from baseline in Eczema Area and Severity Index (EASI) score at week 8 (EASI-75 response, the key secondary end point) was 40.3% in the tapinarof group and 4.3% in the vehicle group (p < 0.0001). In ZBB4-2, IGA treatment success rate was 28.1% at week 16, 32.3% at week 24, and 41.3% at week 52, and EASI-75 response rate was 53.3% at week 16, 63.7% at week 24, and 76.6% at week 52, indicating that efficacy responses improved over time and were maintained over 52 weeks. Across the two trials, most adverse events (AEs) were mild or moderate; common AEs included folliculitis, acne, and headache. In summary, tapinarof cream 1% was effective and generally safe for up to 52 weeks of treatment in Japanese patients with AD.

In vitro killing effect of berberine and niclosamide on ocular Demodex folliculorum

PurposeTo explore the in vitro killing effect of water-soluble berberine and lipid-soluble niclosamide against ocular Demodex folliculorum. MethodsDemodex with good vigor were collected from patients’ eyelashes. These mites were randomly distributed into different groups with 20 mites in each group. Saline, Double Distilled Water (DDW), Polysorbate 80 (TWEEN 80), Polyethylene glycol 300 (PEG 300) and Castor Oil were used to screen solvents and cosolvents. 20 % Tea Tree Oil (TTO) and Anhydrous Ethanol (EtOH) were used as positive controls. 0.2 % Berberine, 0.25 % Niclosamide and 0.5 % Niclosamide, were designated as experimental groups. Following treatment, the analysis of Kaplan-Meier survival curves and survival time of mites and safety of drugs were then performed. ResultsThe survival of Demodex in vitro in Saline and DDW, was not significant different. Therefore, DDW, which was more conducive to the dissolution of berberine, was chosen as the solvent for berberine. 0.2 % Berberine significantly inhibited the survival distribution and survival time (P < 0.001) of Demodex in vitro compared with the DDW group. Through the evaluation of several cosolvents, PEG300 had milder effects on Demodex. Hence, the proportion of PEG300 in the niclosamide solvent group was increased to reduce the irritability of the vehicle. Furthermore, niclosamide could significantly inhibit the survival of Demodex compared with the vehicle group, and the effect of 0.5 % Niclosamide was more obvious (P < 0.001), and was better than 20 %TTO (P < 0.001). In addition, after niclosamide administration, Demodex bodies exhibited gradual distortion along with increased transparency and the presence of blurred dark particles compared to those in the vehicle group. Moreover, both drugs showed good subjective tolerability and safety in a mouse model. Conclusion0.2 % berberine and 0.5 % niclosamide effectively inhibited Demodex survival in vitro, with 0.5 % niclosamide superior to 20 % TTO. These two drugs, with anti-Demodex, anti-bacterial, and anti-inflammatory properties, may offer alternative treatment for Demodex blepharitis.

Mitigatory antioxidant effect of Lycopene against the polystyrene microplastics-induced neurotoxicity via modulation of mTOR/Beclin-1 activities in adult male Wistar rats

BackgroundPolystyrene microplastics (PS-MPs) pose a significant threat to aquatic ecosystems, causing neuronal toxicity in aquatic species, and Lycopene (LYC), a neuroprotective substance, could potentially mitigate this issue. ObjectivesThis study examined the neurotherapeutic benefit of lycopene on the neurochemical status of rats affected by PS-MPs. MethodsTwenty Wistar rats were divided into four groups of five rats each: a vehicle group, a PS-MPs treatment group, a PS-MPs + LYC co-treatment group, and an LYC supplementation group. The experiment lasted 56 days, and all substances were administered orally. Neuronal toxicity was evaluated using oxidative stress/neurochemical biomarkers. Lipid peroxidation (LPO), glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) were measured as indicators of oxidative stress. Nitric Oxide (NO), gamma-aminobutyric acid (GABA), hydroxyl radical (●OH), ammonia, and glutamate levels were assessed to evaluate neurotransmitter balance. Cathepsin marker enzyme activity was also measured to assess lysosomal function. ResultsPS-MPs-treated animals showed increased LPO, NO, ●OH, Bcl-2 interacting protein 1 (BECLIN1), ammonia, glutamate, and reduced GABA, Mammalian targets of rapamycin (mTOR), and antioxidant activity in their cerebral cortex. The combination of PS-MPs and Lycopene treatment improved GABA, cathepsin marker enzyme activity, and brain structure, restoring normal neuronal bodies, oval nuclei, and cerebral cortex. Lycopene ameliorated oxidative stress deficits, decreased LPO, NO, ●OH, ammonia, and glutamate levels, and restored antioxidant enzyme activity. ConclusionIn conclusion, the findings suggest a potential link between PS-MPs and cognitive dysfunction through neurochemical alterations, while the administration of lycopene mitigated the neurochemical changes brought on by PS-MPs.

Cannabidiol Modulates Neuroinflammatory Markers in a PTSD Model Conducted on Female Rats.

Post-traumatic stress disorder (PTSD) is a debilitating neuropsychiatric condition closely linked to neuroinflammation, with a higher prevalence in women. Cannabidiol (CBD), a non-psychoactive cannabinoid, has shown promise as a potential treatment for PTSD. In this study, we used a PTSD model in which female rats were subjected to a severe foot shock followed by contextual situational reminders (SRs). Testing was conducted one month after exposure. The rats received daily CBD injections for three weeks during the SRs, from days 7 to 28. Two days after the final SR, the rats underwent five extinction trials, followed by the forced swim test (FST). After a five-day rest period, the rats were sacrificed, and brain tissues from the medial prefrontal cortex (mPFC) and ventral subiculum (vSUB) were analyzed for inflammatory markers. Chronic CBD treatment reversed impairments in fear extinction caused by shock and SR. It also reduced learned helplessness in the FST and decreased the upregulation of mPFC-il1β induced by shock and SRs. Additionally, exposure to shock and SRs downregulated mPFC-il6 while upregulating vSUB-il6. CBD treatment further downregulated il6 expression in the vSUB compared to the vehicle groups. Our findings show that CBD effectively inhibited the development of PTSD-like behaviors and suppressed neuroinflammation in the mPFC.

Neuroprotective effect of Sigma-2 modulator CT2074 in a mouse model of ocular hypertension

Ocular neurodegenerative diseases, particularly glaucoma, represent a significant global cause of blindness, with current therapies inadequately addressing the degeneration of the retina and optic nerve. Recent research has identified the sigma-2 receptors as a potential druggable target to offer neuroprotection in managing ocular neurodegenerative disorders. This study investigates the neuroprotective potential of CT2074, a sigma-2 receptor modulator, in a mouse model of primary open-angle glaucoma.Male mice were subjected to unilateral magnetic bead-induced elevation of intraocular pressure (IOP) and received daily oral administration of CT2074, commencing three days prior to ocular hypertension (OHT) induction, and continuing for three weeks. Mice received bilateral intraocular injections of cholera toxin B-488 (CTB) to assess retinal ganglion cell (RGC) anterograde transport. Retina, optic nerve, and brain tissues were collected three weeks post OHT induction for quantification of RGC and axon number, with contralateral retinas and cerebelli preserved for assessment of drug exposure.CT2074 was observed in the retina at levels exceeding the 95% receptor occupancy concentration. RGC quantification showed a significant reduction in the Vehicle group compared to Naïve and CT2074 groups. Notably, the CT2074 treatment group exhibited significantly higher RGC density than the Vehicle (p < 0.0001) and was no different than Naïve. Analysis of RGC axons in optic nerve cross-sections revealed significant axonal loss in both the Vehicle and CT2074 groups compared to Naïve, though the CT2074-treated group had significantly higher axon number compared to the Vehicle. Anterograde transport in the Vehicle and CT2074 groups did not differ.This study underscores the potential of CT2074 administered orally to protect RGCs exposed to elevated IOP, as evidenced by substantial preservation of RGCs and their axons compared to Vehicle-treated mice. These findings signify a promising avenue for the development of sigma-2 receptor-targeted therapeutics in glaucoma and related neurodegenerative diseases, addressing a critical unmet need in the field of ocular neuroprotection.

Preclinical Multi-Omic Assessment of Pioglitazone in Skeletal Muscles of Mice Implanted with Human HER2/neu Overexpressing Breast Cancer Xenografts.

Background/Objectives: Breast cancer (BC) is the second most commonly diagnosed cancer worldwide and is accompanied by fatigue during both active disease and remission in the majority of cases. Our lab has measured fatigue in isolated muscles from treatment-naive BC patient-derived orthotopic xenograft (BC-PDOX) mice. Here, we conducted a preclinical trial of pioglitazone in BC-PDOX mice to determine its efficacy in ameliorating BC-induced muscle fatigue, as well as its effects on transcriptomic, metabolomic, and lipidomic profiles in skeletal muscle. Methods: The pioglitazone and vehicle groups were treated orally for 4 weeks upon reaching a tumor volume of 600 mm3. Whole-animal indirect calorimetry was used to evaluate systemic metabolic states. The transcriptome was profiled using short-read bulk RNA sequencing (RNA-seq). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to profile the metabolome and lipidome. Fast and slow skeletal muscle function were evaluated using isolated ex vivo testing. Results: Pioglitazone was associated with a 16.634% lower average O2 consumption (mL∙h-1, p = 0.035), 16.309% lower average CO2 production (mL∙h-1, p = 0.022), and 16.4% lower cumulative energy expenditure (EE) (kcal∙h-1, p = 0.035), with no changes in substrate utilization. RNA-seq supported the downstream effects of pioglitazone on target genes and displayed considerable upregulation of mitochondrial bioenergetic pathways. K-means cluster 5 showed enrichment of the PPAR signaling pathway (adj. p < 0.05, Log2FC = 2.58). Skeletal muscle metabolomic and lipidomic profiles exhibited dysregulation in response to BC, which was partially restored in pioglitazone-treated mice compared to vehicle-treated BC-PDOX mice. In particular, the overall abundance of total ceramide levels was significantly lower in the PioTx group (-46.327%, p = 0.048). Despite molecular support for pioglitazone's efficacy, isolated muscle function was not affected by pioglitazone treatment. No significant difference in the area under the fatigue curve (AUC) was found between the pioglitazone and vehicle groups (p = 0.596). Conclusions: BC induces multi-omic dysregulation in skeletal muscle, which pioglitazone partially ameliorates. Future research should focus on profiling systemic metabolic dysfunction, identifying molecular biomarkers of fatigue, and testing alternative pioglitazone treatment regimens.

Long-term administration of atorvastatin had no effect on the occurrence and progression of spontaneous hypertensive intracerebral hemorrhage in mice

Abstract Background Intracerebral hemorrhage (ICH) is a catastrophic disease with limited treatment options. Statins and low low-density lipoprotein cholesterol (LDL-C) levels have been reported to be associated with increased risk of ICH. Safety concerns of statins were raised as statins become the cornerstone of anti-atherosclerosis therapy. To date, the effect of statins on the incidence of ICH still remains a controversial topic. Purpose The aim of this study was to investigate the effect of atorvastatin on the occurrence and progression of hypertensive ICH in mice. Methods Male wild-type C57BL/6J mice, 13 months old, were randomly divided into atorvastatin (Ato, n=20) group and vehicle (Veh, n=20) group. The atorvastatin group and vehicle group were given atorvastatin (20 mg/kg) and same volume of 0.5% sodium carboxymethylcellulose by oral gavage for 8 weeks, separately. Angiotensin II (Ang II) and L-NAME were then used to induce hypertensive ICH. The time of stroke symptom onset was documented. Hemorrhage volumes were measured by spectrophotometric hemoglobin assay. Results After 8 weeks of atorvastatin or vehicle administration, serum LDL-C levels were significantly lower in the mice treated with atorvastatin than vehicle controls (0.705 vs. 1.008 mmol/L, P&amp;lt;0.001). The administration of atorvastatin did not show significant effects on the systolic blood pressure (SBP) levels when compared with the vehicle group (164 vs. 166 mmHg, P=0.497) after the induction of Ang II and L-NAME. Hemorrhage was observed in brain morphology, and was further verified by hematoxylin and eosin staining or MRI scans in mice that showed signs of stroke. During the 28-day experimental period, 75% (15/20) of mice in the atorvastatin group developed signs of ICH, and 70% (14/20) of mice in the vehicle group developed in the similar time window. The incidence of hypertensive ICH had no significant difference between atorvastatin and vehicle groups (Log-rank P=0.761). Hemorrhage volumes were also comparable between the two groups (1.462 vs. 1.392 uL, P=0.788). Conclusion Reducing serum LDL-C levels by long-term administration of atorvastatin had no effect on the occurrence and progression of spontaneous hypertensive ICH in middle-aged mice.

The role of cellular senescence in aortic dissection

Abstract Background Aortic dissection (AD) is a catastrophic disease with high mortality. Survived patients frequently suffer from serious complications due to progressive destruction of the aortic walls. Recently we have reported that cell proliferation precedes the inflammatory response which is important in AD development and progression. Since cell proliferation promotes cellular senescence that can induce inflammatory response through SASP (senescence-associated secretary phenotype), we hypothesized that cellular senescence plays a critical role in AD pathogenesis. Purpose To investigate if cellular senescence contributes to AD pathogenesis in mouse AD model. Methods and Results A mouse AD model was created by the continuous infusion of beta-aminopropionitrile and angiotensin II (BAPN+AngII) for 14days. BAPN+AngII infusion induced senescence markers and senescent cells in mouse AD model. Senescent cells, as demonstrated by the expression of senescence-associated beta-galactosidase, were evident in intimal endothelial cells, medial smooth muscle cells, adventitial macrophages, and fibroblasts. Rapamycin, an inhibitor of mTOR pathway, suppressed the expression of senescent cells on the aortic tissue and the senescent marker protein expression. To examine the role of cellular senescence in AD, we orally administrated ABT263 known as "senolytics" that eliminates senescent cells. ABT263 treatment prevented cellular senescence in mouse AD model. The AD mortality of ABT263 group was 35%, which was lower than that of vehicle group (66.7%, P &amp;lt; 0.05 by log-rank test). The severity of AD, as assessed by the lesion length in vehicle group was 33.2 ± 3.1mm, whereas that in ABT263 group was 24.6 ± 1.8mm (P &amp;lt; 0.05). Transcriptome analysis revealed that ABT263 treatment suppressed the immune and inflammatory response in the aorta before AD onset. Quantitative RT-PCR confirmed that ABT263 treatment prevented the induction of p21Cip1, interleukin-6, several chemokines and M1 macrophage marker CD80 and CD86. It is known that Jnk activation induces SASP and accelerates cellular senescence. Western blotting confirmed that ABT263 reduced the ratio of pJnk to total Jnk, which was induced by BAPN + AngⅡ. Moreover, aortic smooth muscle cells（SMCs）remained contractile phenotype in ABT263 group, whereas BAPN+AngII reduced that phenotype in vehicle group. Conclusions Cell proliferation via mTOR pathway causes cellular senescence in mouse AD model. Jnk activation was confirmed in mouse AD model. Cellular senescence contributes to AD progression and death associated with inflammatory responses including SASP, the differentiation to M1 macrophage, and the phenotype change of SMCs.Cellular senescence represents a potential predictor and a therapeutic target for AD.

A Study of Heavy Road Freight Transport in Poland in the Context of the Pursuit of Sustainable Road Transport

The efficiency of road freight transport determines—to a significant degree—the total environmental footprint and the amount of greenhouse gases and other pollutants released into the atmosphere by inland transport. The rate of empty or partially empty vehicles is one of the key metrics for improvement of the environmental performance of freight road transportation. This paper presents the characteristics of road freight transport in Poland on the basis of data collected by weigh-in-motion stations. Data aggregation for environmental analysis represents a novel aspect of the work. Indicators describing the degree of loading, the share of empty vehicles in traffic, and the share of vehicles of maximum permissible total weight in traffic were determined for a representative group of heavy goods vehicles. Based on the representative load factor value (LFA), a classification of the road section into four groups was proposed. The results obtained show a clear differentiation of the values of the indicators analyzed for individual groups and their variability in the three-year period covered by the analysis. The aggregation method presented can be used to identify the nature of the distribution of the weight categories of the heavy goods vehicles and provide input information for targeted analyses relating to sustainable road transport management and environmental protection. Finally, the grouped LFA values were linked with indicators of energy consumption (ECI) and on-road emissions (EI).

Understanding the impact of heart failure treatment on cancer growth

Abstract Background Heart failure (HF) is associated with systemic alterations that extend beyond the cardiovascular system. The dysregulation of physiological pathways induced by HF, encompassing inflammatory or neurohormonal pathways, may foster an environment conducive to cancer growth and the spead of metastases. It is unknown whether these alterations are attenuated when HF is treated, and whether certain therapies for HF may yield more significant effects than others. Purpose This study aims to investigate whether myocardial infarction (MI)-induced left ventricular (LV) dysfunction promotes breast cancer growth and triggers the development of lung metastases. Additionally, it compares the potential impact of different therapeutic strategies for LV dysfunction on cancer development. Methods Myocardial infarction (MI) was induced in 11-week-old female BALB/c mice by ligation of the left anterior descending (LAD) coronary artery, leading to LV dysfunction and significant dilation within four weeks. Starting 2 weeks after surgery, mice with ejection fraction (EF) &amp;lt; 40% received daily oral gavage of vehicle or one of three drugs: carvedilol (10 mg/kg), enalapril (20 mg/kg), or empagliflozin (15 mg/kg). Four weeks post-surgery, 2x105 4T1 metastatic breast cancer cells were injected into the 4th mammary fat pad. Tumor volume was measured every two days, starting eight days after injection. Weekly echocardiographic assessments monitored cardiac function until the end of the experiment. Tumors were excised on day 22 after injection. Mice were euthanised on day 28 for post-mortem analysis. MCM2 proliferation staining was performed on lung tissue to quantify metastases. Results Among the HF treatments, only empagliflozin (n=11) induced a significant decrease in both left ventricular end-diastolic and end-systolic volume (Fig. 1.A; p=0.0043; Fig. 1.B; p=0.0186) compared to the vehicle group (n=11). Tumor weight and volume (Fig. 1.C-D) were not different in the HF/vehicle group (n=15) and the sham-operated group (n=13). Carvedilol (n=14) led to a small reduction in tumor weight (Fig. 1.C; p=0.1264), while empagliflozin (n=11) exhibited a significant reduction in both tumor weight (Fig. 1.C; p=0.0306) and volume (Fig. 1.D; p=0.0268). In addition, carvedilol (n=13) significantly reduced lung metastases (Fig. 1.E; p=0.0347) compared to the HF/vehicle group (n=13). Enalapril (n=10) and empagliflozin (n=9) did not significantly impact the development of lung metastases. Conclusion Despite the absence of increased cancer growth post-MI, our findings indicate that specific treatments for HF—namely, carvedilol and empagliflozin—attenuate cancer growth post-MI. Furthermore, carvedilol demonstrated potential in reducing metastatic spread. Additional research is required to uncover the underlying mechanisms both in the presence and absence of HF, and to explore the molecular impact that HF treatments have on various stages of cancer progression.Figure 1

Complement factor H-related protein 5 alleviates joint inflammation and osteoclast differentiation by disrupting RANK-JNK signaling in collagen antibody-induced arthritis mouse model

BackgroundComplement Factor H-Related protein 5 (CFHR5) belongs to the factor H/CFHR family and regulates the complement system by modulating factor H’s inhibitory activity against C3b. Despite its known role, the impact of CFHR5 on autoimmune arthritis and its relationship to pathophysiological changes in arthritis and bone loss remain unclear. This study aimed to assess the effect of CFHR5 on aggressive osteoclast activity and arthritis using a murine model of collagen antibody-induced arthritis (CAIA). MethodsThe effect of recombinant CFHR5 protein (rCFHR5) on arthritis were evaluated in CAIA. The mice were divided into three group and intraperitoneally treated with rCFHR5, methotrexate (MTX) as positive control or PBS as negative control. In the CAIA mouse model, the rCFHR5-treated group significantly reduced the incidence and clinical arthritis equivalent to the MTX group. Clinical arthritis scores, incidence and body weight were measured, and histological analysis of ankle joints was performed by Hematoxylin and Eosin (H&E) and Safranin O − Fast green (SOFG), Tartrate-resistant acid phosphatase (TRAP) staining and Immunohistochemistry. Moreover, to investigate the rCFHR5 role, we isolated murine osteoclast precursor cells (OCPs) from each group, induced osteoclasts with M−CSF and RANKL, and performed TRAP and F-actin staining. To verify the mechanism, mRNA and protein analyses were performed in OCPs. ResultsHistological examination of ankle joints revealed substantial reductions in synovial hyperplasia, bone marrow inflammation, bone erosion, cartilage destruction and TRAP-positive cells in the rCFHR5 group compared to the vehicle group. The ankle joints of the rCFHR5 group showed markedly decreased expression of proinflammatory cytokines (TNF-α, IL-1β and IL-6). Mechanically, treatment with rCFHR5 inhibited RANKL-mediated osteoclast differentiation from OCPs and disrupted the RANK-JNK signaling. These findings demonstrate that treatment with rCFHR5 attenuates joint inflammation and reduces osteoclast differentiation, indicating its potential anti-inflammatory effect in autoimmune arthritis models.

Valproic acid alleviates total-body irradiation-induced small intestinal mucositis in mice

Purpose Gastrointestinal (GI) injury is one of the serious problems of total-body irradiation (TBI). However, no fundamental treatment for TBI and other radiation-induced GI injury has yet been established. Valproic acid (VPA) administration reduces mortality in mice subjected to total-body irradiation (TBI) with X-rays. This study aimed to evaluate the effects of VPA on GI injury induced by TBI in mice. Materials and methods Mice were subjected to TBI with X-rays to induce GI injury. Changes in survival and weight were observed after VPA administration. The small intestine was then sampled at 0, 1, 3, 7, and 10 d after irradiation for histological and immunohistological evaluation and measurement of myeloperoxidase (MPO) activity and inflammatory cytokine levels (IL-1β). Results VPA (200 and 600 mg/kg) increased survival rate and reduced weight loss in model mice. IL-1β expression 1 d after irradiation was significantly lower in the VPA group than that in the vehicle group. Furthermore, the increase in MPO activity at 3 and 7 d after irradiation was significantly suppressed by VPA administration. Histological examination (hematoxylin and eosin staining) revealed that 600 mg/kg VPA inhibited inflammatory cell infiltration. Immunostaining for the proliferating cell nuclear antigen involved in cell proliferation showed that VPA suppressed the irradiation-induced decrease in cell proliferative capacity. Conclusions Treatment with VPA in mice with GI injury caused by TBI suppressed inflammatory responses in small intestinal mucosal cells. These results suggest that VPA may be a useful therapeutic agent against TBI-induced small intestinal mucositis.

Dynamic clustering based risk aware congestion control technique for vehicular network

In order to improve road safety and offer extra services to cars and their users, vehicular ad hoc networks, or VANETs, are essential parts of intelligent transportation systems (ITS). The primary aim of this research is to suggest and assess a new approach for mitigating network congestion in VANET technology through the implementation of dynamic grouping of vehicles for safety (DGVS). To do this, virtual regions are created around cars using the DBSCAN and K Mean method. This allows vehicles to communicate directly with others within the same DGVS, eliminating the need to broadcast messages across the entire network.The ultimate goal is to drastically decrease traffic while preserving vital data transfer for VANET traffic control and safety. The suggested technique determines the optimal transmission rate in accordance with the existing channel circumstances, yielding a balanced performance concerning both packet delivery and channel congestion. With this novel method, cars may only talk to other vehicles that are in the same DGVS, thus there’s no need to broadcast messages to the whole network. With the use of this technique, the efficacy of DGVS in reducing VANET congestion and enhancing network performance will be thoroughly assessed. The study’s conclusions demonstrate how well the suggested dynamic grouping of vehicles for safety (DGVS) technique works to reduce VANET congestion. It was shown through simulation-based studies that, in comparison to current approaches, DGVS dramatically decrease network congestion, resulting in notable gains in network performance and decreased communication delay. Additionally, the study found a number of possible uses for DGVS in the transportation industry, such as emergency response, traffic management, and accident prevention.

Dopamine loss alters glutamate synapses and transporters in the medial prefrontal cortex and anxiety-related behaviour in a male MPTP rodent model of Parkinson's disease.

Anxiety is a prominent non-motor symptom of Parkinson's disease (PD). Changes in the B-spectrum recordings in PD patients of the prefrontal cortex correlate with increased anxiety. Using a rodent model of PD, we reported alterations in glutamate synapses in the striatum and substantia nigra following dopamine (DA) loss. We hypothesize that DA loss will result in increased anxiety-related behaviours and that this will be associated with alterations in glutamate synapses and transporters within the medial prefrontal cortex (mPFC). Following 4weeks of progressive 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration, there was an increase in anxiety-related behaviours and a 78% decrease in plasma corticosterone levels versus the vehicle (VEH)-treated mice. This was associated with a 30% decrease in the density of dendritic spines in Layers Il/Ill, and a 53% decrease in the density of glutamate immuno-gold labelling within vesicular glutamate transporter 1 (Vglut1)-labelled nerve terminals and spines, with no change within vesicular glutamate transporter 2 (Vglut2) positive terminals/spines in the MPTP versus VEH groups. Our prior work determined that a decrease in striatal glutamate terminal density was associated with an increase in extracellular glutamate levels. There was an increase in protein expression of Vglut1 (40%), Vglut2 (37%) and glutamate aspartate transporter (GLAST) (225%), and a decrease in glutamate transporter 1 (GLT-1) (50%) and excitatory amino acid carrier 1 (EAAC1) (51%), in the MPTP versus VEH groups within the mPFC. These data suggest that the decrease in dendritic spines within the mPFC following nigrostriatal DA loss may be due to increased extracellular glutamate levels (decrease in glutamate transporters), leading to an increase in anxiety-related behaviours.

Isoorientin Improves Excisional Skin Wound Healing in Mice

Background/Objectives: Wound healing relies on a coordinated process with the participation of different mediators. Natural products are a source of active compounds with healing potential. Isoorientin is a natural flavone recognized as having several pharmacological properties, such as anti-inflammatory effects, making it a potential treatment for wounds. We investigated the effect of isoorientin on the healing of excisional skin wounds. Methods: Male Swiss mice were subjected to the induction of excisional skin wounds (6 mm diameter) and treated with a vehicle (2% dimethyl sulfoxide in propylene glycol) or 2.5% isoorientin applied topically once a day for 14 days. The wound area was measured on days 0, 3, 7, and 14. Histopathological analyses were performed on the cicatricial tissue after 14 days. The myeloperoxidase activity and the interleukin-1β, tumoral necrosis factor (TNF)-α, and interleukin-6 concentrations were determined on the third day. Results: We observed that 3 days after the topical application of isoorientin, the lesion area was significantly smaller when compared to those of the vehicle (p &lt; 0.01) and control (p &lt; 0.05) groups. No difference was observed after 7 and 14 days of induction. Despite this, on day 14, histological analysis of cicatricial tissue from the animals treated with isoorientin showed reduced epidermal thickness (p &lt; 0.001) and increased collagen deposition (p &lt; 0.001). These effects were accompanied by decreased myeloperoxidase activity and interleukin-1β concentration on the third day of induction, without alteration in TNF-α and interleukin-6. Conclusions: The treatment with isoorientin promoted better tissue repair in excisional wounds in mice, which may be linked to the modulation of the early inflammatory response.

Inhibiting the soluble epoxide hydrolase increases the EpFAs and ERK1/2 expression in the hippocampus of LiCl-pilocarpine post-status epilepticus rat model

PurposeThis study aimed to investigate the enzyme activity of soluble epoxide hydrolase (sEH) and quantify its metabolic substrates, namely epoxygenated fatty acids (EpFAs), and products of sEH in the hippocampus after administering TPPU [1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea], an inhibitor of sEH. Furthermore, it explored whether the extracellular signal-activated protein kinase 1/2 (ERK1/2) is involved in the anti-seizure effects of TPPU in the lithium chloride (LiCl)-pilocarpine induced post-status epilepticus (SE) rat model. MethodsThe rats were intraperitoneally (I.P.) injected with LiCl and pilocarpine to induce SE and then spontaneous recurrent seizures (SRS) were observed. Rats were randomly assigned into SRS + TPPU group (intragastrically administering 0.1 mg/kg/d TPPU), SRS + Vehicle group (administering the vehicle instead), and Control group. Enzyme-linked immunosorbent assay, Western-blot analysis, and ultra-high-performance liquid chromatography/mass spectrometry (LC/MS) were performed to measure the enzyme activity of sEH, the protein level of sEH and ERK1/2, and the concentration of TPPU and polyunsaturated fatty acids (PUFAs) metabolisms in the hippocampus. ResultsThe frequency of SRS events of Racine stage 3 or higher ranged from 0 to 19 per week in the SRS + Vehicle group, compared to 0–5 per week in the SRS + TPPU group. sEH enzyme activity and protein levels were significantly elevated in the SRS + Vehicle group compared to the Control group. After TPPU administration, the hippocampal TPPU concentration reached 10.94 ± 4.37 nmol/kg. sEH enzyme activity was significantly reduced in the LiCl-pilocarpine-induced post-SE rat model, although sEH protein levels did not decrease significantly. The regioisomers 8,9-, 11,12-, and 14,15-EETs, total EETs, the EETs/DHETs ratio, other EpFAs including 16(17)-EpDPA, and the 19(20)-EpDPA/19,20-DiHDPA ratio in the hippocampus were significantly increased. Additionally, the p-ERK1/2 to ERK1/2 ratio in the hippocampus was significantly elevated following TPPU administration. ConclusionThis study demonstrates that inhibiting sEH with TPPU increases the levels of EETs, other EpFAs, and ERK1/2 expression in the hippocampus of a LiCl-pilocarpine-induced post-SE rat model. These findings suggest that the anti-seizure effect of TPPU may be mediated through the EETs-ERK1/2 pathway.

Striking long-term beneficial effects of single dose psilocybin and psychedelic mushroom extract in the SAPAP3 rodent model of OCD-like excessive self-grooming.

Obsessive compulsive disorder (OCD) is a highly prevalent disorder that causes serious disability. Available treatments leave 40% or more of people with OCD significantly symptomatic. There is an urgent need for novel therapeutic approaches. Mice that carry a homozygous deletion of the SAPAP3 gene (SAPAP3 KO) manifest a phenotype of excessive self-grooming, tic-like head-body twitches and anxiety. These behaviors closely resemble pathological self-grooming behaviors observed in humans in conditions that overlap with OCD. Following a preliminary report that the tryptaminergic psychedelic, psilocybin, may reduce symptoms in patients with OCD, we undertook a randomized controlled trial of psilocybin in 50 SAPAP3 KO mice (28 male, 22 female). Mice that fulfilled inclusion criteria were randomly assigned to a single intraperitoneal injection of psilocybin (4.4 mg/kg), psychedelic mushroom extract (encompassing the same psilocybin dose) or vehicle control and were evaluated after 2, 12, and 21 days by a rater blind to treatment allocation for grooming characteristics, head-body twitches, anxiety, and other behavioral features. Mice treated with vehicle (n = 18) manifested a 118.71 ± 95.96% increase in total self-grooming (the primary outcome measure) over the 21-day observation period. In contrast, total self-grooming decreased by 14.60 ± 17.90% in mice treated with psilocybin (n = 16) and by 19.20 ± 20.05% in mice treated with psychedelic mushroom extract (n = 16) (p = 0.001 for effect of time; p = 0.0001 for time × treatment interaction). Five mice were dropped from the vehicle group because they developed skin lesions; 4 from the psilocybin group and none from the psychedelic mushroom extract group. Secondary outcome measures such as head-body twitches and anxiety all showed a significant improvement over 21 days. Notably, in mice that responded to psilocybin (n = 12) and psychedelic mushroom extract (n = 13), the beneficial effect of a single treatment persisted up to 7 weeks. Mice initially treated with vehicle and non-responsive, showed a clear and lasting therapeutic response when treated with a single dose of psilocybin or psychedelic mushroom extract and followed for a further 3 weeks. While equivalent to psilocybin in overall effect on self-grooming, psychedelic mushroom extract showed superior effects in alleviating head-body twitches and anxiety. These findings strongly justify clinical trials of psilocybin in the treatment of OCD and further studies aimed at elucidating mechanisms that underlie the long-term effects to alleviate excessive self-grooming observed in this study. Prepared with BioRender ( https://www.biorender.com/ ).

Application of XBP1s Decoy Oligodeoxynucleotide Attenuates Cancerous Phenotype in Huh-7 Hepatocellular Carcinoma Cells.

The spliced form of X-box binding protein 1 (XBP1s) is a key transcription factor in the unfolded protein response (UPR), an adaptive mechanism for cell survival. Many studies demonstrated the induced expression of XBP1s in various cancers, including hepatocellular carcinoma (HCC). Such upregulated expression is linked to an enhancement of cell proliferation, migration, and improvement of the survival rate. In this study, we aimed to assess the therapeutic potential of targeting XBP1s, by specific decoy oligodeoxynucleotide (ODN) and evaluated the cancerous phenotypes in Huh-7 cells. In this experimental study, we transfected Huh-7 cells with XBP1s decoy oligonucleotide (ODN). Subsequently, we assess some cellular features, including viability, migration capacity, proliferation potential, and apoptosis. Therefore, various techniques included wound healing test, BrdU, and annexin/PI assays. Additionally, the colony formation capacity was evaluated. The mRNA expression levels of BAX, BCL-2, c-MYC, CCND1, MMP-9, CDH1, and CD133 were quantified by the reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Transfection of Huh-7 cells by XBP1s decoy ODN led to significant down-regulation of c-Myc, CCND1, MMP-9, BCL-2 and CD133 and up-regulation of CDH1 and BAX transcriptional expressions in comparison with the vehicle group. Our results also demonstrated that transfection of XBP1s-decoy reduced HCC cell viability, proliferation, migration capacity as well as colonization ability in comparison with the vehicle group. These findings proposed the potential application of XBP1s-decoy ODN to reduce cancerous phenotypes such as cell proliferation, cell migration and apoptosis induction in the Huh-7 cell line. More experiments on other cell lines and primary cells could validate our results.