Experimental Model Research Articles

Creating Fuzzy Models from Limited Data

The design of experiments is a methodological approach in which measurement experiments are carefully planned to obtain highly informative data. This paper addresses the challenge of constructing mathematical models for complex nonlinear processes when the available measurement data have low information content. This problem often arises when data are collected without the guidance of an experimental modeling expert. We examine two practical examples to illustrate this issue: a textile wastewater decolorization process and atmospheric corrosion of structural metal materials. In both cases, the measured data were insufficient to construct highly accurate models. It is, therefore, necessary to make a trade-off between model complexity and accuracy by adapting modeling techniques to work effectively with the limited data available. The main aim of the paper is, therefore, to focus on simple but effective techniques that allow as much information as possible to be extracted from low-quality measurements and to maximize the usefulness of the model for its intended purpose.

Sex similarities and divergences in systemic and muscle iron metabolism adaptations to extreme physical inactivity in rats.

Previous data in humans suggest that extreme physical inactivity (EPI) affects iron metabolism differently between sexes. Our objective was to deepen the underlying mechanisms by studying rats of both sexes exposed to hindlimb unloading (HU), the reference experimental model mimicking EPI. Eight-week-old male and female Wistar rats were assigned to control (CTL) or hindlimb unloading (HU) conditions (n=12/group). After 7days of HU, serum, liver, spleen, and soleus muscle were removed. Iron parameters were measured in serum samples, and ICP-MS was used to quantify iron in tissues. Iron metabolism genes and proteins were analysed by RT-qPCR and Western blot. Compared with control males, control females exhibited higher iron concentrations in serum (+43.3%, p<0.001), liver (LIC; +198%, P<0.001), spleen (SIC; +76.1%, P<0.001), and transferrin saturation (TS) in serum (+53.3%, P<0.001), contrasting with previous observations in humans. HU rat males, but not females, exhibited an increase of LIC (+54% P<0.001) and SIC (+30.1%, P=0.023), along with a rise of H-ferritin protein levels (+60.9% and +134%, respectively, in liver and spleen; P<0.05) and a decrease of TFRC protein levels (-36%; -50%, respectively, P<0.05). HU males also exhibited an increase of splenic HO-1 and NRF2 mRNA levels, (p<0.001), as well as HU females (P<0.001). Concomitantly to muscle atrophy observed in HU animals, the iron concentration increased in soleus in females (+26.7, P=0.004) while only a trend is observed in males (+17.5%, P=0.088). In addition, the H-ferritin and myoglobin protein levels in soleus were increased in males (+748%, P<0.001, +22%, P=0.011, respectively) and in females (+369%, P<0.001, +21.9%, P=0.007, respectively), whereas TFRC and ferroportin (FPN) protein levels were reduced in males (-68.9%, P<0.001, -76.8%, P<0.001, respectively) and females (-75.9%, P<0.001, -62.9%, P<0.001, respectively). Interestingly, in both sexes, heme exporter FLVCR1 mRNA increased in soleus, while protein levels decreased (-39.9% for males P=0.010 and -49.1% for females P<0.001). Taken together, these data support that, in rats (1) extreme physical inactivity differently impacts the distribution of iron in both sexes, (2) splenic erythrophagocytosis could play a role in this iron misdistribution. The higher iron concentrations in atrophied soleus from both sexes are associated with a decoupling between the increase in iron storage proteins (i.e., ferritin and myoglobin) and the decrease in levels of iron export proteins (i.e., FPN and FLVCR1), thus supporting an iron sequestration in skeletal muscle under extreme physical inactivity.

Interaction between Intestinal Parasites and the Gut Microbiota: Implications for the Intestinal Immune Response and Host Defence.

Intestinal parasites, including helminths and protozoa, account for a significant portion of the global health burden. The gastrointestinal (GI) tract not only serves as the stage for these parasitic infections but also as the residence for millions of microbes. As the intricacies of the GI microbial milieu continue to unfold, it is becoming increasingly apparent that the interactions between host, parasite, and resident microbes help dictate parasite survival and, ultimately, disease outcomes. Across both clinical and experimental models, intestinal parasites have been shown to impact microbial composition and diversity. Reciprocally, microbes can directly influence parasitic survival, colonization and expulsion. The gut microbiota can also indirectly impact parasites through the influence and manipulation of the host. Studying this host-parasite-microbiota axis may help bring about novel therapeutic strategies for intestinal parasitic infection as well as conditions such as inflammatory bowel disease (IBD). In this review, we explore the relationship between intestinal parasites, with a particular focus on common protozoa and helminths, and the gut microbiota, and how these interactions can influence the host defence and intestinal immune response. We will also explore the impact of this tripartite relationship in a clinical setting and its broader implications for human health.

Cardioprotective role of long-term kefir and omega-3 fatty acid supplementation on myocardial apoptosis via oxidative stress-mediated lysosomal cathepsin release in isoproterenol-induced myocardial infarction rat model

Objective: The antiapoptotic and antioxidative role of long-term kefir and omega-3 fatty acids and their relationship with cysteine proteases on isoproterenol (ISO) induced myocardial infarction (MI) experimental model was investigated in our study. Material and Methods: Fifty male Sprague-Dawley rats were evenly divided into five distinct groups (n=10): Control, MI, kefir +MI, omega-3+MI, and kefir+omega 3+MI groups. Kefir 10% (with drinking water) and omega-3 fatty acid (30 mg/day per 100g body weight into the standard chow) were administrated during 30 days. ISO was subcutaneously injected into the rats (100 mg/ kg b.w.) on the 29th and 30th days. Myocardial tissue and blood samples were taken 12 hours after the last ISO dose. Creatine kinase MB (CK-MB) activities were measured in serum samples. Caspase-3, superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), DNA fragmentation, cathepsin B and L levels, were measured in myocardial tissue. Results: Serum CK-MB (p&lt;0.05) and cardiac tissue MDA (p&gt;0.05), NO (p&lt;0.01), caspase 3 (p&lt;0.01), DNA fragmentation (p&lt;0.001), cathepsin B (p&lt;0.05) and L (p&lt;0.05) activities were increased and SOD (p&lt;0.001) activities were decreased in MI group compared to control group. The preventive effects of long-term therapy with kefir and omega-3 fatty acids have been demonstrated on apoptosis, oxidative stress markers, and cysteine protease enzymes. Conclusion: Our results showed that long-term administration of kefir and omega-3 fatty acids might be effective in reducing myocardial apoptosis through oxidative stress-mediated release of cysteine proteases in myocardial infarction, especially in the kefir and combined therapy groups.

Human coronavirus OC43 infection remodels connexin 43-mediated gap junction intercellular communication in vitro.

β-coronaviruses cause acute infection in the upper respiratory tract, resulting in various symptoms and clinical manifestations. OC43 is a human β-coronavirus that induces mild clinical symptoms and can be safely studied in the BSL2 laboratory. Due to its low risk, OC43 can be a valuable and accessible model for understanding β-coronavirus pathogenesis. One potential target for limiting virus infectivity could be gap junction-mediated communication. This study aims to unveil the status of cell-to-cell communications through gap junctions in human β-coronavirus infection. Infection with OC43 leads to reduced expression of Cx43 in A549, a lung epithelial carcinoma cell line. Infection with this virus also shows a significant ER and oxidative stress increase. Internal localization of Cx43 is observed post-OC43 infection in the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) region, which impairs the gap junction communication between two adjacent cells, confirmed by Lucifer yellow dye transfer assay. It also affects hemichannel formation, as depicted by the EtBr uptake assay. Impairment of Cx43 trafficking and the ability to form hemichannels and functional GJIC are hampered by virus-induced Golgi apparatus disruption. Altogether, these results suggest that several physiological changes accompany OC43 infection in A549 cells and can be considered an appropriate model system for understanding the differences in gap junction communication post-viral infections. This model system can provide valuable insights for developing therapies against human β-coronavirus infections.IMPORTANCEThe enduring impact of the recent SARS-CoV-2 pandemic underscores the importance of studying human β-coronaviruses, advancing our preparedness for future coronavirus infections. As SARS-CoV-2 is highly infectious, another human β-coronavirus OC43 can be considered an experimental model. One of the crucial pathways that can be considered is gap junction communication, as it is vital for cellular homeostasis. Our study seeks to understand the changes in Cx43-mediated cell-to-cell communication during human β-coronavirus OC43 infection. In vitro studies showed downregulation of the gap junction protein Cx43 and upregulation of the endoplasmic reticulum and oxidative stress markers post-OC43 infection. Furthermore, HCoV-OC43 infection causes reduced Cx43 trafficking, causing impairment of functional hemichannel and GJIC formation by virus-mediated Golgi apparatus disruption. Overall, this study infers that OC43 infection reshapes intercellular communication, suggesting that this pathway may be a promising target for designing highly effective therapeutics against human coronaviruses by regulating Cx43 expression.

Experimental and prediction model of axial compressive responses of corroded RC cylinders strengthened with CFRP

Currently, the capacity of corroded RC cylinders has been proven to significantly improve after being strengthened with carbon fiber reinforced polymer (CFRP). However, existing experimental data remains insufficient, resulting in few models available for predicting the post-strengthening capacity of corroded cylinder. The current understanding of the mechanism by which corrosion affects the mechanical properties of CFRP strengthened cylinders is also not yet clear. Against this background, this study conducted monotonic axial compression tests on CFRP-strengthened corroded RC cylinders. The experimental results indicate that while the CFRP layer predominantly controls the specimen's load-bearing capacity, the reduction in CFRP rupture strain due to increased corrosion rates should not be overlooked. When the corrosion rate reaches 35 %, the experimentally measured average rupture strain was only 65 % of the theoretical value, marking a 30 % decrease compared to non-corroded specimens. Subsequently, a predictive model for peak stress and strain of CFRP-confined corroded RC cylinders was proposed, which considers the confinement mechanisms of stirrups and CFRP as well as the degradation of CFRP rupture strain. Leveraging the existing test data, a dataset comprising 80 CFRP-strengthened corroded RC cylinder specimens was established to calibrate the key parameters in the proposed prediction model. Comparative analysis with existing models indicates that the peak stress and strain prediction model proposed in this study demonstrated the highest forecasting accuracy (the average errors for peak stress and strain were 14 % and 32.6 %, respectively), while existing other predictive models based on limited datasets suffer from a lack of universality. The experimental data and research conclusions of this article can provide reference for engineering practice and the establishment of models in subsequent research.

Exploring the Therapeutic Potential of Green-Synthesized Gold Nanoparticles and Ericaria selaginoides Extract for Inflammatory Bowel Disease.

Addressing disease remission and treatment adherence in inflammatory bowel diseases (IBDs), such as Crohn's disease, poses significant challenges due to underlying oxidative and inflammatory processes. Nanotechnology emerges as a promising avenue for enhancing therapeutic outcomes in IBD by optimizing drug bioactivity, reducing toxicity, and extending circulation time. Gold nanoparticles, known for their resistance to gastrointestinal pH and possessing antioxidant and anti-inflammatory properties, offer particular promise. They can be produced by green synthesis with seaweed Ericaria selaginoides (ES), itself associated with gastroprotective and anti-inflammatory activities. In a murine model of Crohn's disease induced with 8% acetic acid, pretreatment with dexamethasone (0.2 mL/30 g) or Au@ES (25 and 50 mg/kg) effectively mitigated inflammatory features. Notably, ES (50 mg/kg) and Au@ES (50 mg/kg) administration resulted in significant reductions in both macroscopic and microscopic inflammation scores compared to the disease control group. Furthermore, these treatments normalized inflammatory cytokine expression while safeguarding myenteric plexus glial cells. They also impeded neutrophil activation, leading to reduced myeloperoxidase activity and lipid peroxidation, coupled with increased glutathione levels. In conclusion, ES and Au@ES exhibit potent efficacy in counteracting inflammation and oxidation processes in an experimental Crohn's disease model, suggesting their potential as alternative therapeutic strategies for IBD.

Electrolytic synthesis of γ-Al2O3 nanoparticle from aluminum scrap for enhanced methylene blue adsorption: experimental and RSM modeling

The presence of methylene blue (MB) dye in wastewater has raised concern about human health and environmental ecology due to potential carcinogenic, and mutagenic effects. Therefore, this work aims to remove MB dye from wastewater using γ-Al2O3 nanoparticles synthesized from aluminum scrap via simple electrolytic method. The successful synthesis of the adsorbent was confirmed by a range of spectroscopy and microscopy techniques, including XRD, SEM, FTIR, and BET. The central composite design (CCD) of the response surface methodology (RSM) method was used to optimize the processing parameters such as solution pH, contact time, initial MB concentration, and adsorbent dose. The ANOVA results clearly shows that the quadratic model (p < 0.0001) was sufficient to the best predicting of the removal performance of MB dye (R2 = 0.9862). The optimum condition for the maximum MB dye removal (98.91%) was achieved at solution pH of 8.298, initial MB concentration of 31.657 mg/L, adsorbent dose of 0.387 g/L, and contact time of 46.728 min. Nano-γ-Al2O3 was shown to have a good surface area of 59 mg2/g by BET analysis. The adsorption kinetics follows the pseudo-second-order model (R2 = 0.997). With a maximum adsorption capacity of 137.17 mg/g, the Langmuir isotherm model (R2 = 984) provides the best fit to the adsorption isotherm data, indicating a monolayer adsorption process. Furthermore, thermodynamic analysis demonstrated that the adsorption of MB dye was an endothermic and spontaneous process. The reusability study showed that γ-Al2O3 nano-adsorbent retained 85.08% of its original removal efficiency after five cycles. According to the findings of the study, MB dye molecules were taken up by γ-Al2O3 nano-adsorbent via hydrogen bond formation, Van der Waals interaction, and electrostatic attraction. Therefore, γ-Al2O3 nanoparticles can be used as a potentially eco-friendly and low-cost adsorbent for the removal of MB dye from aqueous solutions.

Comprehensive Experimental Database and Model Fitting for Electric Arc Behavior in Aircraft Environments

The More Electric Aircraft (MEA) paradigm advocates weight reduction by transitioning to electrical components, resulting in increased system voltage and higher arc fault risk. This paper presents a methodology for creating a database of nearly one thousand tests simulating parallel arc faults, using Andrea's arc model for accurate representation. Objectives include the creation of a robust experimental database and the use of a simplified model to analyze arcing behavior. The setup follows the UL 746A standard and considers various pressure and electrode separation conditions. The database includes 960 experiments with PVC and PTFE materials, and model fitting ensures accurate representation of arc currents and voltages. Error analysis shows consistent model performance across materials and highlights sensitivity to electrode spacing, pressure, and source voltage. The paper concludes with a clear 30% error threshold for model validity, enhancing the understanding of the applicability of Andrea's model under various experimental scenarios in aircraft environments.

Dietary supplementation of black cumin (Nigella sativa) meal in the formulation of protein‐enriched cookies, further in vivo evaluation of protein quality with physicochemical and organoleptic characterization

AbstractNigella sativa has solid historical importance as a medicinal herb and is widely used in the bakery industry. The study revealed that the defatted black cumin meal (DBCM) possesses a high nutritious composition and amino acid profile, such as Leu, Lys, Arg, Phe, and Glu. The supplementation effect of wheat flour with defatted black cumin meal (DBCM) at levels of 5%–25% was explored on the cookies' nutritional, functional, and organoleptic attributes. Moreover, the present work was conducted to assess the in vivo protein quality of defatted black cumin meal (DBCM)‐supplemented cookies using the albino rat's experimental modeling for 10 days. Rheological characterization suggested a solid elastic behavior for all cookies dough with elevation in G' (138,652–230,926 Pa), G" (34201–45,092 Pa), and G* values due to protein addition. The functional characteristics of the flour blends showed that DBCM addition significantly (p ≤ .05) improved oil (70.56%–165%) and water (84.83%–232.67%) absorption capacities, emulsion, and foaming stability. The physicochemical and organoleptic assessment of the cookies exposed that a 20% level of substitution of DBCM produced acceptable cookies. The net protein utilization (NPU), protein efficiency ratio (PER), true digestibility (TD), and biological value (BV) significantly differ from each other among all the supplementation diet groups with 0%–25% DBCM.

Efficacy and Safety Assessment of Topical Omega Fatty Acid Product in Experimental Model of Irritant Contact Dermatitis: Randomized Controlled Trial

Contact dermatitis is a common inflammatory skin disease that often requires prescription therapy and is associated with adverse reactions. Omega fatty acids have been recognized for their anti-inflammatory effect and could serve as a safer option in contact dermatitis treatment. Therefore, the aim of this randomized controlled study, conducted at the University of Split School of Medicine, was to evaluate the efficacy and safety of omega fatty acids containing topical products in an experimental model of irritant contact dermatitis. This study was registered with ClinicalTrials (NCT06189144) and is closed. The primary outcomes were levels of transepidermal water loss, skin hydration, and skin erythema, all measured using an MPA6 device in 25 healthy participants. A significant difference was observed between the hydration values of the intervention (45.7 ± 12.4) and control groups (31.6 ± 12.3) (p &lt; 0.05) on final measurements (day 10). Moreover, higher erythema levels were observed in participants who were smokers, compared to non-smokers. No adverse drug reactions were observed during the study period. In conclusion, omega fatty acids topical product use shows promise in the treatment of irritant contact dermatitis, and further studies are needed to evaluate efficacy in a larger sample of patients.

Gingivo-Periosteal Expansion of Edentulous Jaw Crest with An Osmotic Self-Inflatable Expander: A Preclinical in Vivo Study.

This study examined the biocompatibility and expansion volume of tissue expanders utilizing rabbits and beagles as experimental models. The self-inflatable expander was provided using a Tissue balloon® (Neobiotech Co., Seoul, Korea). In 9 adult rabbits, a Tissue balloon® was placed under the lateral mandibular periosteum via an extraoral approach. After 2, 3, and 4 weeks (n=3), the expanders were removed, and soft tissue and bone samples were collected for analysis. Histomorphometric analysis and measurements of expander characteristics were performed. In 5 Beagles, all premolars were extracted. Three months after extraction, tissue expanders were placed in the maxilla and mandible, adjacent to dental extraction sites, and removed after 3 weeks. Gingival parameters were measured before and after expansion. Blood perfusion in the oral mucosa was assessed using a laser Doppler flowmeter at different time points. After three weeks, dogs were euthanized, and tissue samples were collected for histologic analysis, focusing on inflammatory response, bone formation, and gingival thickness changes. In the rabbit study, measurements of size, weight, and volume showed significant increases over 4 weeks. Swelling peaked at 2 weeks postimplantation and then gradually stabilized. Histologic examination revealed no signs of inflammation, and expanders were covered by collagen-rich capsules. Some bone resorption was noted due to pressure from the expanders, but the resorption was not significant. In the beagle study, twenty tissue expanders were implanted; 1 expander was lost, and 9 wounds dehisced during soft tissue healing. However, no signs of infection were noted. Histologic evaluation showed bone atrophy, attributed to swelling pressure, at the recipient site. Gingival thickness was not decreased. Perfusion measurements indicated that expansion did not affect microvascular circulation adversely. Overall, both studies suggest that tissue expanders demonstrate biocompatibility and successful expansion volume in vivo, with minimal adverse effects on surrounding tissues and microvascular circulation.

Studies on the thermal sensitivity of lung cancer cells exposed to an alternating magnetic field and magnesium-doped maghemite nanoparticles

BackgroundMagnetic fluid hyperthermia (MFH) represents a promising therapeutic strategy in cancer utilizing the heating capabilities of magnetic nanoparticles when exposed to an alternating magnetic field (AMF). Because the efficacy and safety of MFH treatments depends on numerous intrinsic and extrinsic factors, therefore, the proper MFH setups should focus on thermal energy dosed into the cancer cells.MethodsIn this study, we performed MFH experiments using human lung cancer A549 cells (in vitro) and NUDE Balb/c mice bearing human lung (A549) cancer (in vivo). In these two experimental models, the heat was induced by magnesium-doped iron(III) oxide nanoparticles coated with mPEG-silane (Mg0.1-γ-Fe2O3(mPEG-silane)0.5) when exposed to an AMF.ResultsWe observed that the lung cancer cells treated with Mg0.1-γ-Fe2O3(mPEG-silane)0.5 (0.25 mg·mL−1) and magnetized for 30 min at 14.4 kA·m−1 yielded a satisfactory outcome in reducing the cell viability up to ca. 21% (in vitro). The activation energy calculated for this field strength was estimated for 349 kJ·mol−1. Both volumetric measurements and tumor mass assessments confirmed by magnetic resonance imaging (MRI) showed a superior thermal effect in mice bearing human lung cancer injected intratumorally with Mg0.1-γ-Fe2O3(mPEG-silane)0.5 nanoparticles (3 mg·mL−1) and subjected to an AMF (18.3 kA·m−1) for 30 min four times at weekly intervals. Research demonstrated that mice undergoing MFH exhibited a marked suppression of tumor growth (V = 169 ± 94 mm3; p < 0.05) in comparison to the control group of untreated mice. The CEM43 (cumulative number of equivalent minutes at 43 °C) value for these treatments were estimated for ca. 9.6 min with the specific absorption rate (SAR) level ranging from 100 to 150 W·g−1.ConclusionsThe as-obtained results, both cytotoxic and those related to energy calculations and SAR, may contribute to the advancement of thermal therapies, concurrently indicating that the proposed magnetic fluid hyperthermia holds a great potential for further testing in the context of medical applications.Graphical

Female Mice Exposed to Pyriproxyfen Since Prepuberty Showed Reproductive Impairment During Sexual Maturity and Increased Fetal Death in Their Offspring.

Pyriproxyfen (PPF) is an insecticide used in agriculture, which is approved for use in drinking water tanks for human consumption. However, some studies indicate that it may act as an endocrine disruptor and affect nontarget organisms. This study aimed to evaluate the effects of PPF on reproduction and general health status in female mice exposed from pre-puberty to adulthood. In the first experiment, females were treated by gavage from postnatal day (PND) 23 to (PND) 75 and were distributed into three experimental groups: control (vehicle), PPF 0.1 mg/kg, and PPF 1 mg/kg. Female mice were assessed for the age of puberty onset, body mass, water and food consumption, and the estrous cycle. On PDN 75, a subgroup was euthanized, when vital and reproductive organs were collected and weighed. The thyroid, ovary, and uterus were evaluated for histomorphometry. The other subgroup was assessed in relation to reproductive performance and fetal parameters. In a second experiment, the uterotrophic assay was performed with juvenile females (PND 18) using doses of 0.01, 0.1, or 1 mg/kg of PPF. PPF treatment reduced thyroid mass and increased liver mass. Furthermore, there was an increase in ovarian interstitial tissue and, in the uterus, a decrease in the thickness of the endometrial stroma with reduced content of collagen fibers. There was also a reduction of 30% in pregnancy rate in the treated groups and an increase in the frequency of fetal death. This study suggests that, based on this experimental model, the insecticide may pose a reproductive risk for females chronically exposed to the substance from the pre-pubertal period until adulthood. These results raise concerns about prolonged exposure of women to the same compound.

Multiple Administration of Dexamethasone Possesses a Deferred Long-Term Effect to Glycosylated Components of Mouse Brain.

Glucocorticoids are used during glioblastoma treatment to prevent the cerebral edema effect surrounding normal brain tissue. The aim of our study was to investigate the long-term effects of multiple administrations of glucocorticoids onto the glycosylated components (proteoglycans and glycosaminoglycans) of normal brain extracellular matrix and the glucocorticoid receptor (GR, Nr3c1) in an experimental model in vivo. Two-month-old male C57Bl/6 mice (n = 90) were injected intraperitoneally with various doses of dexamethasone (DXM) (1; 2.5 mg/kg) for 10 days. The mRNA levels of the GR, proteoglycans core proteins, and heparan sulfate metabolism-involved genes were determined at the 15th, 30th, 60th, and 90th days by a real-time RT-PCR. The glycosaminoglycans content was studied using dot blot and staining with Alcian blue. A DXM treatment increased total GAG content (2-fold), whereas the content of highly sulfated glycosaminoglycans decreased (1.5-2-fold). The mRNA level of the heparan sulfate metabolism-involved gene Hs3St2 increased 5-fold, the mRNA level of Hs6St2 increased6-7-fold, and the mRNA level of proteoglycan aggrecan increased 2-fold. A correlation analysis revealed an association between the mRNA level of the GR and the mRNA level of 8 of the 14 proteoglycans-coding and 4 of the 13 heparan sulfate metabolism-involved genes supporting GR involvement in the DXM regulation of the expression of these genes. In summary, multiple DXM administrations led to an increase in the total GAG content and reorganized the brain extracellular matrix in terms of its glycosylation pattern.

Investigation of Convective Heat Transfer and Stability on a Rotating Disk: A Novel Experimental Method and Thermal Modeling

Experimental and numerical investigations are conducted on a rotating disk from the perspective of convective heat transfer to understand the effect of heating on the stability of flow. A non-invasive approach with a thermal camera is employed to determine local Nusselt numbers for different rotational rates and perturbation parameters, i.e., the strength of the heat transfer. A novel transient temperature data extraction over the disk radius and an evaluation method are developed and applied for the first time for the air on a rotating disk. The evaluation method utilizes the lumped capacitance approach with a constant heat flux input. Nusselt number distributions from this experimental study show that there is a good agreement with the previous experimental correlations and linear stability analysis on the subject. A significant result of this approach is that by using the experimental setup and developed approach, it is possible to qualitatively show that instability in the flow starts earlier, i.e., an earlier departure from laminar behavior is observed at lower rotational Reynolds numbers with an increasing perturbation parameter, which is due to the strength of heating. Two experimental setups are modeled and simulated using a validated in-house Python code, featuring a three-dimensional thermal model of the disk. The thermal code was developed for the rotating disks and brake disks with a simplified geometry. Experimentally evaluated heat transfer coefficients are implemented and used as convective boundary conditions in the thermal code. Radial temperature distributions are compared with the experimental data, and there is good agreement between the experiment and the model. The model was used to evaluate the effect of radial conduction, which is neglected when using the lumped capacitance approach to determine heat transfer coefficients. It was observed that the radial conduction has a slight effect. The methodology and approach used in this experimental study, combined with the numerical model, can be used for further investigations on the subject.

Biological and Procedural Predictors of Outcome in the Stroke Preclinical Assessment Network (SPAN) Trial.

The SPAN trial (Stroke Preclinical Assessment Network) is the largest preclinical study testing acute stroke interventions in experimental focal cerebral ischemia using endovascular filament middle cerebral artery occlusion (MCAo). Besides testing interventions against controls, the prospective design captured numerous biological and procedural variables, highlighting the enormous heterogeneity introduced by the multicenter structure that might influence stroke outcomes. Here, we leveraged the unprecedented sample size achieved by the SPAN trial and the prospective design to identify the biological and procedural variables that affect experimental stroke outcomes in transient endovascular filament MCAo. The study cohort included all mice enrolled and randomized in the SPAN trial (N=1789). Mice were subjected to 60-minute MCAo and followed for a month. Thirteen biological and procedural independent variables and 4 functional (weight loss and 4-point neuroscore on days 1 and 2, corner test on days 7 and 28, and mortality) and 3 tissue (day 2, magnetic resonance imaging infarct volumes and swelling; day 30, magnetic resonance imaging tissue loss) outcome variables were prospectively captured. Multivariable regression with stepwise elimination was used to identify the predictors and their effect sizes. Older age, active circadian stage at MCAo, and thinner and longer filament silicone tips predicted higher mortality. Older age, larger body weight, longer anesthesia duration, and longer filament tips predicted worse neuroscores, while high-fat diet and blood flow monitoring predicted milder neuroscores. Older age and a high-fat diet predicted worse corner test performance. While shorter filament tips predicted more ipsiversive turning, longer filament tips appeared to predict contraversive turning. Age, sex, and weight interacted when predicting the infarct volume. Older age was associated with smaller infarcts on day 2 magnetic resonance imaging, especially in animals with larger body weights; this association was most conspicuous in females. High-fat diet also predicted smaller infarcts. In contrast, the use of cerebral blood flow monitoring and more severe cerebral blood flow drop during MCAo, longer anesthesia, and longer filament tips all predicted larger infarcts. Bivariate analyses among the dependent variables highlighted a disconnect between tissue and functional outcomes. Our analyses identified variables affecting endovascular filament MCAo outcome, an experimental stroke model used worldwide. Multiple regression refuted some commonly reported predictors and revealed previously unrecognized associations. Given the multicenter prospective design that represents a sampling of real-world conditions, the degree of heterogeneity mimicking clinical trials, the large number of predictors adjusted for in the multivariable model, and the large sample size, we think this is the most definitive analysis of the predictors of preclinical stroke outcome to date. Future multicenter experimental stroke trials should standardize or at least ensure a balanced representation of the biological and procedural variables identified herein as potential confounders.

Impact of ionic strength on goethite colloids co-transported with arsenite (As3+) through a saturated sand column under anoxic condition: Experiment and mathematical modeling

The knowledge about co-transport of goethite and As3+ to investigate the effect of goethite colloids on As3+ transport under various degrees of seawater intrusion, particular extremely conditions, in groundwater environment is still limited. The main objective is to investigate the influence of seawater intrusion on the sorption, migration, and reaction of As3+and goethite colloids into sand aquifer media under anoxic conditions by using the bench-scale and reactive geochemical modeling. The research consisted of two parts as follows: 1) column transport experiments consisting of 8 columns, which were packed by using synthesis groundwater at IS of 0.5, 50, 200, and 400 mM referring to the saline of seawater system in the study area, and 2) reactive transport modeling, the mathematical model (HYDRUS-1D) was applied to describe the co-transport of As3+ and goethite. Finally, to explain the interaction of goethite and As3+, the Derjaguin-Landau-Verwey-Overbeek (DLVO) calculation was considered to support the experimental results and HYDRUS-1D model. The results of column experiments showed goethite colloids can significantly inhibit the mobility of As3+ under high IS conditions (>200 mM). The Rf of As3+ bound to goethite grows to higher sizes (47.5 and 65.0 μm for 200 and 400 mM, respectively) of goethite colloid, inhibiting As3+ migration through the sand columns. In contrast, based on Rf value, goethite colloids transport As3+ more rapidly than a solution with a lower IS (0.5 and 50 mM). The knowledge gained from this study would help to better understand the mechanisms of As3+ contamination in urbanized coastal groundwater aquifers and to assess the transport of As3+ in groundwater, which is useful for groundwater management, including the optimum pumping rate and long-term monitoring of groundwater quality.

The emerging roles of necroptosis in skeletal muscle health and disease.

Necroptosis is a regulated form of cell death with implications in various physiological and pathological processes in multiple tissues. However, the relevant findings from post-mitotic tissues, such as skeletal muscle, are scarce. This review summarizes the potential contributions of necroptosis to skeletal muscle health and diseases. It first discusses the physiological roles of necroptosis in muscle regeneration and development. It then summarizes the contributions of necroptosis to the pathogenesis of multiple muscle diseases, including muscular dystrophies, inflammatory myopathies, cachexia, and neuromuscular disorders. Lastly, it unravels the gaps in our understanding and therapeutic challenges of inhibiting necroptosis as a potential intervention for muscle diseases. Specifically, the findings from the transgenic animal models and the use of pharmacological inhibitors of necroptosis are discussed with relevance to improving the structure and/or function of skeletal muscle in various diseases. Recent developments from experimental animal models and clinical data are presented to discuss the roles of necroptosis in skeletal muscle health and diseases.

A preclinical mice model of multiple sclerosis based on the toxin-induced double-site demyelination of callosal and cerebellar fibers

BackgroundMultiple sclerosis (MS) is an irreversible progressive CNS pathology characterized by the loss of myelin (i.e. demyelination). The lack of myelin is followed by a progressive neurodegeneration triggering symptoms as diverse as fatigue, motor, locomotor and sensory impairments and/or bladder, cardiac and respiratory dysfunction. Even though there are more than fourteen approved treatments for reducing MS progression, there are still no cure for the disease. Thus, MS research is a very active field and therefore we count with different experimental animal models for studying mechanisms of demyelination and myelin repair, however, we still lack a preclinical MS model assembling demyelination mechanisms with relevant clinical-like signs.ResultsHere, by inducing the simultaneous demyelination of both callosal and cerebellar white matter fibers by the double-site injection of lysolecithin (LPC), we were able to reproduce CNS demyelination, astrocyte recruitment and increases levels of proinflammatory cytokines levels along with motor, locomotor and urinary impairment, as well as cardiac and respiratory dysfunction, in the same animal model. Single site LPC-injections either in corpus callosum or cerebellum only, fails in to reproduce such a complete range of MS-like signs.ConclusionWe here report that the double-site LPC injections treatment evoke a complex MS-like mice model. We hope that this experimental approach will help to deepen our knowledge about the mechanisms of demyelinated diseases such as MS.