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Objectives. The aim was to demonstrate a reliable ex vivo method to test the function of the whole heart. Design. Pigs of varying sizes (44–80 kg) were exposed to dose response of adrenaline. Blood pressures and cardiac output were measured. The explanted hearts were tested in a novel ex vivo system to see if we could replicate the in vivo values at maximal adrenaline stimulation. The perfusion solution was STEEN Solution™ with erythrocytes and continuous infusion of essential drugs. In contrast to normal body circulation which is sequential, the heart evaluation system is divided into left and right heart circuits which are operating in parallel, making it possible to test the right and left heart individually or as a whole. The system provides coronary flow measurements. The nonlinear dynamic resistances are constructed to stabilize systolic and diastolic pressures in a broad range and independently from cardiac output. It is important for the functional evaluation to avoid pumping help for the heart; therefore, atrial vortexes are constructed to minimize pump flow directionality and energy from entering atria. Results. Ex vivo evaluation was able to match the maximal in vivo effect of adrenaline on cardiac output and blood pressures. After 2 h of evaluation, the blood gases and lactate were normal and free haemoglobin was zero. Autopsy of the hearts showed no macroscopic pathology. Conclusions. The system is able to give a reliable functional evaluation of the heart ex vivo. [ABSTRACT FROM AUTHOR]

Mucopolysaccharidosis IVA (MPS IVA) is an autosomal recessive disease caused by a mutation in the N-acetylgalactosamine-6-sulfate-sulfatase (GALNS) gene resulting in progressive systemic skeletal dysplasia. There is currently no effective treatment available for this skeletal condition. Thus, the development of a new therapy stands as an unmet challenge in reversing or alleviating the progression of the disease. Our research, which could be a game-changer, hypothesizes that ex vivo lentiviral (LV) gene therapy (GT) could produce the supraphysiological level of active GALNS enzyme by hematopoietic stem cells (HSCs) transduced with LVs carrying the native GALNS gene under two different promoters (CBh and COL2A1), impacting bone and cartilage abnormalities in MPS IVA. We conditioned newborn knock-out (Galns−/−) MPS IVA mice with busulfan and intravenously transplanted LV-modified HSCs isolated from the bone marrow of Galns−/− donor mice. Transplanted mice were autopsied at 16 weeks, and tissues were collected to assess the therapeutic efficacy of modified HSCs in MPS IVA mice. Although HSC-LV-CBh-hGALNS provided a higher GALNS enzyme activity in plasma, HSC-LV-COL2A1-hGALNS stably corrected heart and bone abnormalities better under a low level of GALNS enzyme. Our findings suggest that ex vivo LV-GT may potentially treat MPS IVA. [ABSTRACT FROM AUTHOR]

Background: Infections caused by biofilm-forming bacteria have significantly linked to dental plaque and caries. The aim of this study is to assess efficacy of some natural compounds in inhibition and eradication of biofilm formed by bacterial isolates from dental infections. Results: Bacterial isolates were recovered from dental plaque/caries and identified using standard microbiological tests and 16S rDNA sequencing. The isolated bacterial strains include Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Klebsiella pneumoniae, and Escherichia coli. The antibiotic susceptibility was determined by disk diffusion method and revealed that the majority of isolates showed high antibiotic resistance, and 61% of isolates were found to be multidrug resistant. The biofilm formation capacity of isolates was investigated using microtiter plate assay. Among the 77 bacterial isolates, seventeen showed moderate biofilm formation capacity, twenty-two showed near-moderate, thirty-four had weak biofilm-forming capacity, and four were non-biofilm producers. The antibiofilm activity of tested compounds (rose and jasmine oils, propolis, vanillin, and vinegar) was evaluated against isolates with highest biofilm-forming capacity. The in vitro antibiofilm ability of tested substances were investigated alone or in combination with each other to evaluate their ability to prevent biofilm formation or destroy preformed single-/multispecies biofilms. Finally, antibiofilm ability of tested combination was evaluated ex vivo on natural teeth. Our results showed that vanillin in combination with rose or jasmine oils showed promising biofilm inhibition and biofilm eradication activities in both the in vitro and ex vivo models. Conclusions: Dental plaque and caries can be successfully prevented using combination of vanillin with rose or jasmine oils, and these compounds can be incorporated in new anticaries dental formulations. [ABSTRACT FROM AUTHOR]

Abstract Purpose Mutations in ARID1A frequently occur in colorectal cancer (CRC) cells. However, there are currently no clinical treatment options specifically addressing this aberration. The preliminary in vitro experiments revealed a synthetic lethal interaction between ARID1A and Aurora kinase A (AURKA) in colorectal cancer (CRC) cells. Methods We collected samples from 80 CRC patients and evaluated the efficacy of AURKA inhibitor (AURKAi) using the ATP-tumor chemosensitivity assay (ATP-TCA) on untreated ARID1A-proficient (ARID1A +) and ARID1A-deficient (ARID1A-) CRC patient samples. In addition, we validated this result by a clonogenic assay. Additionally, we examined the effects of AURKA inhibitors on cell cycle progression and apoptosis in ARID1A + and ARID1A- CRC patient samples using flow cytometry. Results The results showed that AURKAi selectively inhibited the growth of ARID1A- CRC cells. Furthermore, AURKA inhibitors significantly increased G2/M arrest and induced apoptosis in ARID1A- cells. Conclusion We believe that AURKAi hold promise as potential therapeutics for ARID1A mutation colorectal cancer patients.

Purpose: To compare the myelin water fraction (MWF) measurements between 3 T and 7 T and between in vivo and ex vivo human brains, and to investigate the relationship between multi‐echo gradient‐echo (mGRE)‐based 3D MWF and myelin content using histological staining, which has not been validated in the human brain. Methods: In this study, we performed 3D mGRE‐based MWF measurements on five ex vivo human brain hemispheres and five healthy volunteers at 3 T and 7 T with 1 mm isotropic resolution. The data were fitted with the T2*$$ {\mathrm{T}}_2^{\ast } $$ based on a three compartment complex‐valued model to estimate MWF. We obtained myelin basic protein (MBP) staining from two tissue blocks and co‐registered the MWF map and histology image for voxel‐wise correlation between the two. Results: The MWF values measured from 7 T were overall higher than 7 T, but data between the two field strength demonstrated high correlations both in vivo (r = 0.88) and ex vivo (r = 0.83) across 19 white matter regions. Moreover, the MWF measurements showed a good agreement between in vivo and ex vivo assessments at 3 T (r = 0.61) and 7 T (r = 0.54). Based on MBP staining, the MWF values exhibited strong positive correlations with myelin content on both 3 T (r = 0.68 and r = 0.78 for the two tissue blocks) and 7 T (r = 0.64 and r = 0.82 for the two tissue blocks). Conclusion: The findings demonstrated that the mGRE‐based MWF mapping can be used to quantify myelin content in the human brain, despite the field‐strength dependency of the measurements. [ABSTRACT FROM AUTHOR]

In this study, we observed the effect of the newly isolated probiotic strain Limosilactobacillus reuteri B1/1 on the relative gene expression of selected cytokines (interleukin-15, transforming growth factor-β4), tight junction proteins (E-cadherin, occludin), biomarker active intestinal stem cells - LGR5 (leucine-rich repeat containing G protein-coupled receptor), markers of mucosal intestinal immunity (mucin-2, immunoglobulin A), as well as the creation of a new biomarker of inflammation in the intestine - calprotectin on an ex vivo model of chicken ileal explant in the prevention of Salmonella Enteritidis PT4 infection. The ability of L. reuteri B1/1 to effectively modulate the mucosal immune response under pretreatment conditions in S. Enteritidis PT4 infection in a chicken ileal explant model was confirmed. In addition, our obtained results point to the fact that the new chicken ileum explant model could be a suitable model to investigate or test the influence of natural substances such as probiotic bacteria in the interaction with the intestine as well as pathogenic microorganisms. In addition, the results of our study may contribute to a deeper understanding of the action of newly isolated probiotic bacteria at the intestinal level using ex vivo models such as chicken ileum explant, which are able to mimic in vivo conditions sufficiently. [ABSTRACT FROM AUTHOR]

Objective: To evaluate anti-psoriatic activity of Coleus forskohlii in rats with imiquimod-induced psoriasis. Methods: Imiquimod was used to induce psoriasis in rats. Body weight, skin thickness, erythema, scaling, spleen weight, and histological alternations were measured to assess the effect of Coleus forskohlii. Furthermore, an emulgel formulation containing Coleus forskohlii 10% was prepared and characterized along with its ex vivo permeation studies. Results: The emulgel formulation containing Coleus forskohlii 10% had a pH of 5.40±0.36, with optimum spreadability of (31.67±2.08) g/(cms) and viscosity of (15 966.67±1 274.10) cps, and enhanced both the rate and the extent of drug permeation through psoriatic skin. In an ex vivo study, the quantity of drug permeated (19.18%), deposited (52.38%), and drug remaining in the donor compartments (28.31%) was satisfactory. Coleus forskohlii significantly alleviated imiquimod-induced psoriasis by increasing glutathione and superoxide dismutase activity, decreasing malondialdehyde and nitric oxide levels, and alleviating histological alternations in rat skin. Conclusions: Coleus forskohlii can alleviate imiquimod-induced psoriasis, which may be used as a therapeutic candidate for the treatment of psoriasis.

In this paper, we report on the use of a purpose-built hybrid solid-state microwave and radiofrequency generator operating at frequencies of 2.45 GHz and/or 480 kHz for cancer ablation in various tissues. The hybrid generator was tested ex vivo on chicken breast and bovine liver and has demonstrated that the high accuracy of the power delivered to the sample can be achieved by controlling the emitted power versus the temperature profile of the treated sample. In particular, the hybrid generator incorporates control systems based on impedance or reflected power measurements that allow controlled ablation without causing unwanted carbonization and without including areas where tissue damage is not desired. The results of the ex vivo tests showed that radiofrequency ablation (RFA) could be effective for performing controlled ablations with minimally invasive probes, such as cardiac pathologies, small lesions, and tissues with particular composition, while microwave ablation (MWA) could be optimal for performing large ablations in highly vascularized tissues, such as liver cancer, where it is necessary to achieve higher temperatures. [ABSTRACT FROM AUTHOR]

Objective: To evaluate anti-psoriatic activity of Coleus forskohlii in rats with imiquimod-induced psoriasis. Methods: Imiquimod was used to induce psoriasis in rats. Body weight, skin thickness, erythema, scaling, spleen weight, and histological alternations were measured to assess the effect of Coleus forskohlii. Furthermore, an emulgel formulation containing Coleus forskohlii 10% was prepared and characterized along with its ex vivo permeation studies. Results: The emulgel formulation containing Coleus forskohlii 10% had a pH of 5.40±0.36, with optimum spreadability of (31.67±2.08) g/(cms) and viscosity of (15 966.67±1 274.10) cps, and enhanced both the rate and the extent of drug permeation through psoriatic skin. In an ex vivo study, the quantity of drug permeated (19.18%), deposited (52.38%), and drug remaining in the donor compartments (28.31%) was satisfactory. Coleus forskohlii significantly alleviated imiquimod-induced psoriasis by increasing glutathione and superoxide dismutase activity, decreasing malondialdehyde and nitric oxide levels, and alleviating histological alternations in rat skin. Conclusions: Coleus forskohlii can alleviate imiquimod-induced psoriasis, which may be used as a therapeutic candidate for the treatment of psoriasis. [ABSTRACT FROM AUTHOR]

Acanthamoeba is a ubiquitous genus of amoebae that can trigger a severe and progressive ocular disease known as Acanthamoeba Keratitis (AK). Furthermore, current treatment protocols are based on the combination of different compounds that are not fully effective. Therefore, an urgent need to find new compounds to treat Acanthamoeba infections is clear. In the present study, we evaluated staurosporine as a potential treatment for Acanthamoeba keratitis using mouse cornea as an ex vivo model, and a comparative proteomic analysis was conducted to elucidate a mechanism of action. The obtained results indicate that staurosporine altered the conformation of actin and tubulin in treated trophozoites of A. castellanii. In addition, proteomic analysis of treated trophozoites revealed that this molecule induced overexpression and a downregulation of proteins related to key functions for Acanthamoeba infection pathways. Additionally, the ex vivo assay used validated this model for the study of the pathogenesis and therapies of AK. Finally, staurosporine eliminated the entire amoebic population and prevented the adhesion and infection of amoebae to the epithelium of treated mouse corneas. [ABSTRACT FROM AUTHOR]

Human precision-cut lung slices (hPCLS) prepared from fibrotic lungs recapitulate the pathophysiological hallmarks of fibrosis. These hallmark features can also be induced by treating non-fibrotic hPCLS with a fibrotic cocktail (FC). As a result, the fibrotic and fibrosis-induced hPCLS are rapidly emerging as preferred models for disease modeling and drug discovery. However, current hPCLS models are limited by tissue viability in culture, as they are usually only viable for one week after harvesting. Here, we demonstrate that the fibrotic hPCLS can be cryopreserved, stored for months, and then thawed on demand without loss of hPCLS viability or protein content for 14 days post-thawing. Cryopreservation also preserves the pro-fibrotic potential of non-fibrotic hPCLS. Specifically, when we treated the thawed non-fibrotic hPCLS with an FC, we observed significant pro-fibrotic cytokine secretion and elevated tissue stiffness. These pro-fibrotic changes were inhibited by the small-molecule tyrosine kinase inhibitor, Nintedanib. Taken together, our work indicates that a feasible solution to prolong the pre-clinical utility of fibrotic and fibrosis-induced hPCLS is cryopreservation. We anticipate that cryopreserved hPCLS will serve as an advantageous predictive model for the evaluation of pro-fibrotic pathways during acute and chronic toxicity testing. [ABSTRACT FROM AUTHOR]

The goal of the present study was to prepare an in situ carmustine gel to evaluate the viability of brain targeting via nasal administration. Initial formulations (CG1–CG5) were made, and their varied medicinal properties were assessed. Polymers that were employed as mucoadhesive with thermoreversible materials were carbopol 934 and poloxamer 407 respectively. The gel formulations were made using a cold process. Using a visual technique, the phase transition temperature was determined. The in vitro release, ex vivo carmustine permeability via goat nasal mucosal membrane, pH, mucoadhesive strength, and in vivo pharmacokinetic studies of the gels were assessed. The purpose of the histological analysis of the nasal mucosal membrane was to look for signs of harmful effects caused by carmustine permeability. It was discovered that the 18.00 percentage w/v poloxamer mixture was exhibiting a phase change at physiological temperatures (34.00 to 35.00 °C). The gelling temperature was found to drop when the carbopol 934 proportion was raised as of 0.1– 0.5 percentage weight/volume. The mucoadhesive strength of every preparation was more than 3500 dynes/cm2. A study on carmustine permeation has shown that employing carbopol 934 at concentrations higher than 0.3% w/v can accelerate the rate of carmustine permeation. No indication of injury has been found in the histological investigation of the nasal mucosa following the carmustine permeation trial. Compared to commercial intravenous (IV) injections of carmustine, in vivo pharmacokinetic analysis of carmustine in situ gel revealed increased Cmax, AUC0→t, and AUC0→∞ and lower Tmax (1 h) data in the brain. Hence, in situ thermoreversible mucoadhesive intranasal gel of carmustine might be an impactful therapy in the management of glioblastoma (GBM). [ABSTRACT FROM AUTHOR]

Leishmania parasites are transmitted to mammalian hosts through the bite of sandflies. These parasites can infect phagocytic cells (macrophages, dendritic cells, and neutrophils) and non-phagocytic cells (B cells and fibroblasts). In mice models, the disease development or resolution is linked to T cell responses involving inflammatory cytokines and the activation of macrophages with the M1/M2 phenotype. However, this mechanism does not apply to human infection where a more complex immunological response occurs. The understanding of interactions between immune cells during Leishmania infection in humans is still limited, as current infection models focus on individual cell types or late infection using controlled human infection models (CHIMs). This study investigated the early parasite infection in freshly isolated peripheral blood-derived (PBD) leukocytes over 24 h. Flow cytometer analysis is used in immunophenotyping to identify different subpopulations. The study found that among the L. aethiopicaGFP-associated leukocytes, most cells were neutrophils (55.87% ± 0.09 at 4 h) and monocytes (23.50% ± 0.05% at 24 h). B cells were 12.43% ± 0.10% at 24 h. Additionally, 10–20% of GFP+ leukocytes did not belong to the aforementioned cell types, and further investigation revealed their identity as CD4+ T cells. Data not only confirm previous findings of Leishmania infection with PBD leukocytes and association with B cells but also suggest that CD4+ T cells might influence the early-stage of infection. [ABSTRACT FROM AUTHOR]

Background Methods Results Conclusions Ex vivo machine perfusion (EVMP) has been established to extend viability of donor organs. However, EVMP protocols are inconsistent. We hypothesize that there is a significant relationship between specific parameters during EVMP and perfusion outcomes.A meta‐analysis of literature was conducted in accordance with the Preferred Reporting Items for Systematic Review and Meta‐Analysis (PRISMA) Statement. The search encompassed articles published before July 25, 2023. PubMed, Embase, and CENTRAL databases were screened using search terms “ex‐vivo,” “ex‐situ,” “machine,” and “perfusion.” Weight gain, an indicator of organ viability, was chosen to compare outcomes. Extracted variables included perfused organ, warm and cold ischemia time before perfusion, perfusion duration, perfusate flow, pressure, temperature, perfusate composition (presence of cellular or acellular oxygen carrier, colloids, and other supplements) and percent weight change. Data were analyzed using SPSS statistical software.Overall, 44 articles were included. Red blood cell‐based perfusates resulted in significantly lower weight gain compared to acellular perfusates without oxygen carriers (11.3% vs. 27.0%, p < 0.001). Hemoglobin‐based oxygen carriers resulted in significantly lower weight gain compared to acellular perfusates (16.5% vs. 27%, p = 0.006). Normothermic perfusion led to the least weight gain (14.6%), significantly different from hypothermic (24.3%) and subnormothermic (25.0%) conditions (p < 0.001), with no significant difference between hypothermic and subnormothermic groups (24.3% vs. 25.0%, p = 0.952). There was a positive correlation between flow rate and weight gain (ß = 13.1, R = 0.390, p < 0.001).Oxygen carriers, low flow rates, and normothermic perfusate temperature appear to improve outcomes in EVMP. These findings offer opportunities for improving organ transplantation outcomes. [ABSTRACT FROM AUTHOR]

Next‐generation bioelectronic implants require miniaturization, durability, and long‐term functionality. Thin film encapsulations, prepared with inorganic or hybrid organic/inorganic designs, are essential for ensuring protection, low water permeation, adaptability, and structural strength. It is equally important to precisely measure their barrier performance, especially for in vivo use, to ensure the manufacture of reliable bioelectronics. Current monitoring solutions are not adequate: they are bulky, lack sensitivity, and are incompatible with microfabricated devices. Here, a comprehensive method is introduced to quantify the permeability of thin‐film encapsulation for bioelectronic implants both in situ and in real time. This method relies on monitoring the electrical resistance of the Mg film, which experiences corrosion due to water permeation, leading to Mg hydrolysis. An analytical model is proposed that predicts and quantifies this permeation, and is adaptable for various types of encapsulations, including hybrid multilayers. An unprecedented ultra‐low detection limit of 3 × 10−8 g m−2 d−1 at room temperature is demonstrated and the monitoring approach is validated in vivo using polyimide and poly(dimethylsiloxane)‐coated bioelectronics. [ABSTRACT FROM AUTHOR]

Berberine is an isoquinolone alkaloid present in a wide variety of plants. Other than Type 2 diabetes and hypertension it is also found to have an effect in lowering blood cholesterol levels. However, because of its poor oral bioavailability and substantial first pass hepatic metabolism it is consumed in higher dosage leading to side effects. To reduce dosage and avoid adverse effects, transdermal delivery is investigated using transethosomal gel as a carrier. Transethosomes are ethanolic nanocarriers that are specifically designed for the dermal or transdermal delivery route as it has enhanced permeation of the vesicles. In our study we have successfully investigated the potential use of berberine loaded transethosomal gel (BBR-TE) as a carrier for transdermal delivery. Three-level three-factorial Box Behnken Design was used to optimize BBR-TE (Opt-BBR-TE). The optimized formulation was further evaluated and found to have a particle size of 165.70 nm with entrapment efficiency of 79.60±4.07%. The in vitro drug release study suggested the non-Fickian transport mechanism. The ex vivo skin permeation was found to be 94.35±3.64% when compared to berberine conventional formulation. Berberine has also shown promising antioxidant activity when compared to the standard solution and was entrapped inside TE vesicles in non-crystalline form. The Opt-BBR-TE was converted into gel form. Various parameters of the gel such as pH, Drug Content, Spreadability and extrudability were found to have promising results. Present study data revealed that the developed transethosomal gel formulation was found to be a potentially useful drug carrier for transdermal delivery of berberine. [ABSTRACT FROM AUTHOR]

Fatty acid amide hydrolase (FAAH) is a conserved hydrolase in eukaryotes with promiscuous activity toward a range of acylamide substrates. The native substrate repertoire for FAAH has just begun to be explored in plant systems outside the model Arabidopsis thaliana. Here, we used ex vivo lipidomics to identify potential endogenous substrates for Medicago truncatula FAAH1 (MtFAAH1). We incubated recombinant MtFAAH1 with lipid mixtures extracted from M. truncatula and resolved their profiles via gas chromatography–mass spectrometry (GC–MS). Data revealed that besides N‐acylethanolamines (NAEs), sn‐1 or sn‐2 isomers of monoacylglycerols (MAGs) were substrates for MtFAAH1. Combined with in vitro and computational approaches, our data support both amidase and esterase activities for MtFAAH1. MAG‐mediated hydrolysis via MtFAAH1 may be linked to biological roles that are yet to be discovered. [ABSTRACT FROM AUTHOR]

Objectives: To determine the imaging details and diagnostic information of the treatment response to neoadjuvant chemoradiotherapy (nCRT) of rectal adenocarcinoma at 9.4T magnetic resonance imaging (MRI) by ex vivo. Methods: Fifteen cases with locally advanced rectal cancer (LARC) followed by radical surgery after nCRT between September 2022 and February 2023 were recruited. Resected specimens were fixed in a perfluoropolyether‐filled test tube and scanned with a 3.0T and 9.4T MRI system ex vivo. The residual tumor depth and MRI‐based tumor regression grade (TRG) were subjectively assessed and then compared with the pathological findings. Results: The ex vivo 9.4T T2WI without fat suppression clearly differentiated tumor tissue, fibrosis and normal rectal wall, which clearly corresponded to the pathologic tissues of the rectal specimens. The TRG could be accurately assessed on ex vivo 9.4T images in 13/15 specimens (86.7%), while in 11/15 specimens (73.3%) on ex vivo 3.0T images. Conclusion: Ex vivo 9.4T MR imaging clearly displayed the components of rectal wall and proved excellent diagnostic performance for evaluating the treatment response to nCRT, which allow radiologists to understand and then assess more accurately the TRG of LARC after nCRT. [ABSTRACT FROM AUTHOR]

The role of vertical ex vivo dermoscopy relevant to clinical diagnosis has not been investigated yet. Study objectives were defining, describing, and determining the importance of the structures visible using vertical ex vivo dermoscopy in the diagnosis of malignant skin lesions, as well as determining their accuracy in the assessment of tumor margins. A prospective, descriptive study was conducted in two University centers. Digital images of completely excised skin lesions, fixed in formalin, before histopathological diagnosis were used for analysis. BCCs had the most diverse dermoscopic presentation on the vertical section, while SCCs showed a similar presentation in most cases. Vertical dermoscopy of thin melanomas was almost identical, unlike nodular melanomas. Thickness accuracy assessed by dermatologist was 0.753 for BCC, 0.810 for SCC, and 0.800 for melanomas, whereas assessment by pathologist was 0.654, 0.752, and 0.833, respectively. The accuracy of tumor width assessment was 0.819 for BCCs, 0.867 for SCCs and 1.000 for melanoma as estimated by a Dermatologist. Interobserver agreement was 0.71 for BCC, 0.799 for SCC and 0.832 for melanomas. Vertical ex vivo dermoscopy may contribute to the distinction between BCCs, SCCs, and melanomas. Moreover, regardless of the doctor's specialty, it enables a good assessment of the tumor's margins. [ABSTRACT FROM AUTHOR]

Background: This study evaluated electrical conductivity in human liver tissue in the 3–1000 kHz frequency range to compare normal versus tumor tissues under in vivo versus ex vivo conditions. Methods: Previous informed consent was obtained from twenty patients undergoing liver resection in whom liver electrical conductivity was measured during surgery and after resection. Result: We found higher electrical conductivity values in tumor tissues than in normal tissue in both in vivo (0.41 ± 0.10 vs. 0.13 ± 0.06 S/m) and ex vivo (0.27 ± 0.09 vs. 0.12 ± 0.07 S/m) conditions (at 3 kHz). The electric properties also showed a promising potential for distinguishing between different tissue types including metastasis, cholangiocarcinoma (CCA), hepatocellular carcinoma (HCC), hepatic cirrhosis, and normal liver (both in vivo and ex vivo). At 3 kHz, in vivo electrical conductivity for cholangiocarcinoma, HCC, and metastasis were 0.35, 0.42 ± 0.13, and 0.41 ± 0.08 S/m, respectively, which differed significantly from each other (p < 0.05). Conclusions: These findings could potentially improve liver disease diagnostics through electrical conductivity measurements and treatment techniques involving electric fields. Future research should focus on expanding the sample size to refine the categorization and comparison processes across diverse human liver tissue types. [ABSTRACT FROM AUTHOR]

Lung diseases such as cancer substantially alter the mechanical properties of the organ with direct impact on the development, progression, diagnosis, and treatment response of diseases. Despite significant interest in the lung's material properties, measuring the stiffness of intact lungs at sub-alveolar resolution has not been possible. Recently, we developed the crystal ribcage to image functioning lungs at optical resolution while controlling physiological parameters such as air pressure. Here, we introduce a data-driven, multiscale network model that takes images of the lung at different distending pressures, acquired via the crystal ribcage, and produces corresponding absolute stiffness maps. Following validation, we report absolute stiffness maps of the functioning lung at microscale resolution in health and disease. For representative images of a healthy lung and a lung with primary cancer, we find that while the lung exhibits significant stiffness heterogeneity at the microscale, primary tumors introduce even greater heterogeneity into the lung's microenvironment. Additionally, we observe that while the healthy alveoli exhibit strain-stiffening of ~1.75 times, the tumor's stiffness increases by a factor of six across the range of measured transpulmonary pressures. While the tumor stiffness is 1.4 times the lung stiffness at a transpulmonary pressure of three cmH2O, the tumor's mean stiffness is nearly five times greater than that of the surrounding tissue at a transpulmonary pressure of 18 cmH2O. Finally, we report that the variance in both strain and stiffness increases with transpulmonary pressure in both the healthy and cancerous lungs. Our new method allows quantitative assessment of disease-induced stiffness changes in the alveoli with implications for mechanotransduction. [ABSTRACT FROM AUTHOR]

Adipose tissue and skeletal muscle dysfunction play a central role in cardiometabolic morbidity. Ashwagandha and Andrographis are purported to have anti-inflammatory and antioxidant activity, but this is based on exposure of cells to the parent compounds ignoring phytochemical absorption and metabolism. We explored the anti-inflammatory/antioxidant effects of ashwagandha and Andrographis in ex vivo human models of skeletal muscle and adipose tissue. Healthy participants supplemented with 2000 mg/day Andrographis (n = 10) or 1100 mg/day ashwagandha (n = 10) for 28 days. Sera collected pre (D0) and post (D28) supplementation were pooled by timepoint and added to adipose explant (AT) and primary human myotube (SKMC) culture media (15% v/v) for treatment. A Taqman panel of 56 genes was used to quantify these. In AT, treatment with ashwagandha sera decreased the expression of genes involved in antioxidant defence and inflammatory response (CCL5, CD36, IL6, IL10, ADIPOQ, NFEL2, UCP2, GPX3, GPX4; geometric 95% CI for fold change > 1) and altered the expression of genes involved in fatty acid metabolism. In SKMC, ashwagandha sera altered FOXO1 and SREBF1 expression. Andrographis sera decreased IL18 and SERPINEA3 expression in AT. This physiologically relevant in vitro screening characterises the effects of ashwagandha in AT to guide future clinical trials. [ABSTRACT FROM AUTHOR]

In this study, scalp tissues from Korean adults between 20 and 80 without skin disease were used. Scalp tissues were processed, and hair follicles were isolated and cultured with different treatments (including Bioscalp, Ultra Exo Booster, and Ultra S Line Plus) from Ultra V company. Over 12 days, observations and measurements of hair follicle characteristics were recorded at intervals (Days 0, 3, 6, 9, and 12). The study assessed the impact of these substances on hair follicle growth and morphology. Bioscalp, combined with Ultra Exo Booster and Ultra S Line Plus, showed significant hair elongation in ex vivo. Preservation of hair bulb diameter was observed, indicating potential for sustained hair growth by exosome‐based products. The hair growth cycle analysis suggested a lower transition to the catagen stage in test products from Ultra V compared to non‐treated groups. The research findings indicated that the tested formulations, especially the combination of Bioscalp, Ultra Exo Booster, and Ultra S Line Plus, demonstrated significant effectiveness in promoting hair growth, maintaining the integrity of the hair bulb, and reducing the transition to the catagen stage. The study suggests promising alternative treatments for hair loss, illustrating results that were as good as those of the conventional testing product groups. [ABSTRACT FROM AUTHOR]

Deafness-causing deficiencies in otoferlin (OTOF) have been addressed preclinically using dual adeno-associated virus (AAV)-based approaches. However, timing of transduction, recombination of mRNA, and protein expression with dual hybrid AAV methods methods have not previously been characterized. Here, we have established an ex vivo assay to determine the kinetics of dual-AAV mediated expression of OTOF in hair cells of the mouse utricle. We utilized two different recombinant vectors that comprise DB-OTO, one containing the 5′ portion of OTOF under the control of the hair cellspecific Myo15 promoter, and the other the 3′ portion of OTOF. We explored specificity of the Myo15 promoter in hair cells of the mouse utricle, established dose response characteristics of DB-OTO ex vivo in an OTOF-deficient mouse model, and demonstrated tolerability of AAV1 in utricular hair cells. Furthermore, we established deviations from a one-to-one ratio of 5′ to 3′ vectors with little impact on recombined OTOF. Finally, we established a plateau in quantity of recombined OTOF mRNA and protein expression by 14 to 21  days ex vivo with comparable recovery timing to that in vivo model. These findings demonstrate the utility of an ex vivo model system for exploring expression kinetics and establish in vivo and ex vivo recovery timing of dual AAV-mediated OTOF expression. [ABSTRACT FROM AUTHOR]

High resolution 3D printing emerges as an alternative to microfabrication due to its fine resolution along with one-step manufacturing. Thus, it is broadly used in many fields, such as biological and chemical applications. We introduce such a technique to the design of the optofluidic probe by integrating optics and microfluidics as an ex vivo liquid core fiber photometry. We build the optofluidic probes with various T-shapes and conduct the transmission measurements and the ray tracing simulations, where the results are comparable. Through the transmission and fluorescence measurements, we obtain optimized curl T-shape dimensions of 524 µm wide, ∼50 µm thick, and 350 µm long with longitudinal spaces between them of 260 um. Furthermore, a heightened level of complexity in structure, characterized by a feature size of 25 µm, is attained through the improvement process. We conclude the feasibility of this optofluidic system with two applications: the in vivo-like setting consisting of thyroid biopsy training phantom and human plasma and the ex vivo-like setting consisting of the mice brain slices stained with wheat germ agglutinin (WGA). This prototype is an important step of establishing a 3D printing optofluidic applications for various in vivo research.

Ex vivo tissue culture of the human corpus cavernosum (CC) can be used to explore the tissue structural changes and complex signaling networks. At present, artificial CC-like tissues based on acellular or three-dimensional (3D)-printed scaffolds are used to solve the scarcity of primary penis tissue samples. However, inconvenience and high costs limit the wide application of such methods. Here, we describe a simple, fast, and economical method of constructing artificial CC-like tissue. Human CC fibroblasts (FBs), endothelial cells (ECs), and smooth muscle cells (SMCs) were expanded in vitro and mixed with Matrigel in specific proportions. A large number of bubbles were formed in the mixture by vortexing combined with pipette blowing, creating a porous, spongy, and spatial structure. The CC FBs produced a variety of signaling factors, showed multidirectional differentiation potential, and grew in a 3D grid in Matrigel, which is necessary for CC-like tissue to maintain a porous structure as a cell scaffold. Within the CC-like tissue, ECs covered the surface of the lumen, and SMCs were located inside the trabeculae, similar to the structure of the primary CC. Various cell components remained stable for 3 days in vitro, but the EC content decreased on the 7th day. Wingless/integrated (WNT) signaling activation led to lumen atrophy and increased tissue fibrosis in CC-like tissue, inducing the same changes in characteristics as in the primary CC. This study describes a preparation method for human artificial CC-like tissue that may provide an improved experimental platform for exploring the function and structure of the CC and conducting drug screening for erectile dysfunction therapy.

This study investigates the mechanical behavior of femur bones under loading conditions, focusing on the transition from elastic to plastic deformation and eventual fracture. The force–displacement curves reveal distinct phases of deformation, with an initial linear relationship indicating elastic behavior, followed by deviation from linearity marking the onset of plastic deformation. Fracture occurs beyond a critical load, leading to a sharp drop in the force–displacement curve. The maximum fracture force varies among specimens and is influenced by bone geometry, size, cross-sectional area, and cortical thickness. Post-failure analysis highlights additional insights into fracture mechanics and bone material toughness. Reinforcing bones with screws enhances their strength, which is evident in the higher fracture forces observed in force–displacement diagrams. Fixation procedures following fractures further increase bone strength. Comparing specimens with and without strengthening underscores the effectiveness of reinforcement methods in improving bone mechanical properties. After analyzing the results, it is evident that femur bones with reinforcement can withstand greater loads, and they can also absorb higher impact energies while remaining in the elastic deformation range and without suffering permanent plastic damage. This study provides valuable insights into bone biomechanics and the efficacy of reinforcement techniques in enhancing bone strength and fracture resistance. [ABSTRACT FROM AUTHOR]

To investigate the feasibility of using neutron tomography to gain new knowledge of human articular cartilage degeneration in osteoarthritis (OA). Different sample preparation techniques were evaluated to identify maximum intra-tissue contrast. Human articular cartilage samples from 14 deceased donors (18–75 years, 9 males, 5 females) and 4 patients undergoing total knee replacement due to known OA (all female, 61–75 years) were prepared using different techniques: control in saline, treated with heavy water saline, fixed and treated in heavy water saline, and fixed and dehydrated with ethanol. Neutron tomographic imaging (isotropic voxel sizes from 7.5 to 13.5 µm) was performed at two large scale facilities. The 3D images were evaluated for gradients in hydrogen attenuation as well as compared to images from absorption X-ray tomography, magnetic resonance imaging, and histology. Cartilage was distinguishable from background and other tissues in neutron tomographs. Intra-tissue contrast was highest in heavy water-treated samples, which showed a clear gradient from the cartilage surface to the bone interface. Increased neutron flux or exposure time improved image quality but did not affect the ability to detect gradients. Samples from older donors showed high variation in gradient profile, especially from donors with known OA. Neutron tomography is a viable technique for specialized studies of cartilage, particularly for quantifying properties relating to the hydrogen density of the tissue matrix or water movement in the tissue. [ABSTRACT FROM AUTHOR]

Hydatidosis causes a serious health hazard to humans and animals leading to significant economic and veterinary and public health concern worldwide. The present study aimed to evaluate the in vitro and ex vivo protoscolicidal effects of synthesized poly(amidoamine), PAMAM, nanoemulsion. In this study, PAMAM was characterized through dynamic light scattering technique to investigate the particle size and zeta potential of nanoemulsified polymer. For the in vitro and ex vivo assays, we used eosin dye exclusion test and scanning electron microscope (SEM) to evaluate the effects of the prepared and characterized PAMAM nanoemulsion against protoscoleces from Echinococcus granulosus sensu lato G6 (GenBank: OQ443068.1) isolated from livers of naturally infected camels. Various concentrations (0.5, 1, 1.5 and 2 mg/mL) of PAMAM nanoemulsion at different exposure times (5, 10, 20 and 30 min) were tested against protoscolices. Our findings showed that PAMAM nanoemulsion had considerable concentration- and time-dependent protoscolicidal effect at both in vitro and ex vivo experiments. Regarding in vitro assay, PAMAM nanoemulsion had a potent protoscolicidal effect when compared with the control group with a highest protoscolicidal activity observed at the concentration of 2 mg/mL at all exposure times, such that 100% of protoscolices were killed after 20 min of exposure. Also, the mortality of protoscolices was 100% after 30 min of exposure to 1 and 1.5 mg/mL of PAMAM nanoemulsion, in vitro. Concerning ex vivo assay PAMAM nanoemulsion recorded the highest mortality rates at the concentration of 2 mg/mL (55, 99.4 and 100% at 10, 20, 30 min, respectively). Ultrastructure examination of examined protoscolices after 20 min of exposure to PAMAM nanoemulsion showed a complete loss of rostellar hooks, disruption of suckers with disorganization of hooks with partial or complete loss of them, and damage of protoscolices tegument with loss of their integrity in the form of holes and contraction of the soma region were observed in 1.5 and 2 mg/mL of PAMAM, in vitro and ex vivo, showing more damage in the in vitro conditions. It can be concluded that PAMAM nanoemulsion is a promising protoscolicidal agent offering a high protoscolicidal effect at a short exposure time. Further in vivo studies and preclinical animal trials are required to evaluate its efficacy and clinical applications against hydatid cysts. [ABSTRACT FROM AUTHOR]

The economic impact of necrotizing enteritis (NE) resulting from Clostridium perfringens infection has been significant within the broiler industry. This study primarily investigated the antibacterial efficacy of hexahydrocolupulone against C. perfringens, and its pharmacokinetics within the ileal contents of broiler chickens. Additionally, a dosing regimen was developed based on the pharmacokinetic/pharmacodynamic (PK/PD) model specific to broiler chickens. Results of the study indicated that the minimum inhibitory concentration (MIC) of hexahydrocolupulone against C. perfringens ranged from 2mg/L to 16mg/L in MH broth. However, in ileal content, the MIC ranged from 8mg/L to 64mg/L. The mutation prevention concentration (MPC) in the culture medium was found to be 128mg/L. After oral administration of hexahydrocolupulone at a single dosage of 10-40mg/kg bodyweight, the peak concentration (Cmax), maximum concentration time (Tmax), and area under the concentration-time curve (AUC) in ileal content of broiler chickens were 291.42-3519.50 µg/g, 1-1.5h, and 478.99-3121.41 µgh/g, respectively. By integrating the in vivo PK and ex vivo PD data, the AUC0-24h/MIC values required for achieving bacteriostatic, bactericidal, and bacterial eradication effects were determined to be 36.79, 52.67, and 62.71h, respectively. A dosage regimen of 32.9mg/kg at 24h intervals for a duration of 3days would yield therapeutic efficacy in broiler chickens against C. perfringens, provided that the MIC below 4mg/L. [ABSTRACT FROM AUTHOR]

Simple Summary: SCFAs are considered functional ingredients as they promote the growth and health of fish. This work aimed to assess the effect of short-chain fatty acids (SCFAs) on the immune status of the European sea bass gut through the ex vivo use of intestinal explants. The ex vivo technique significantly reduces the number of animals to be used, according to the guidelines of the 3Rs. The present results are important to understand how European sea bass respond to a bacterial infection after the administration of SCFAs and for the evaluation of functional ingredients used in aquaculture. This study aimed to evaluate the intestinal interactions between three short-chain fatty acids (SCFA), namely, acetate, propionate, and butyrate, and pathogenic bacteria (Vibrio anguillarum) in intestinal explants of European sea bass (Dicentrarchus labrax) juveniles. The anterior intestine of 12 fish with an average weight of 100 g (killed by excess anesthesia with 2-phenoxyethanol) were sampled and placed in 24-well plates. The experimental treatments consisted of a control medium and a control plus 1 mM or 10 mM of sodium acetate (SA), sodium butyrate (SB), and sodium propionate (SP). After 2 h of incubation, the explants were challenged with Vibrio anguillarum at 1 × 107 CFU/mL for 2 h. After the bacterial challenge, and regardless of the SCFA treatment, the oxidative stress-related genus catalase (cat) and superoxide dismutase (sod) were down-regulated and glutathione peroxidase (gpx) was up-regulated. Furthermore, the immune-related genes, i.e., the tumor necrosis factor (TNF-α), interleukin 8 (IL-8), transforming growth factor (TGF-β), and nuclear factor (NF-Kβ) were also up-regulated, and interleukin 10 (IL-10) was down-regulated. During the pre-challenge, sodium propionate and sodium butyrate seemed to bind the G-protein coupled receptor (grp40L), increasing its expression. During the challenge, citrate synthase (cs) was down-regulated, indicating that the SCFAs were used as an energy source to increase the immune and oxidative responses. Overall, our results suggest that sodium propionate and sodium butyrate may boost European sea bass immune response at the intestine level. [ABSTRACT FROM AUTHOR]

The genus Senna contains globally distributed plant species of which the leaves, roots, and seeds have multiple traditional medicinal and nutritional uses. Notable chemical compounds derived from Senna spp. include sennosides and emodin which have been tested for antimicrobial effects in addition to their known laxative functions. However, studies of the effects of the combined chemical components on intact human gut microbiome communities are lacking. This study evaluated the effects of Juemingzi (Senna sp.) extract on the human gut microbiome using SIFR® (Systemic Intestinal Fermentation Research) technology. After a 48-hour human fecal incubation, we measured total bacterial cell density and fermentation products including pH, gas production and concentrations of short chain fatty acids (SCFAs). The initial and postincubation microbial community structure and functional potential were characterized using shotgun metagenomic sequencing. Juemingzi (Senna seed) extracts displayed strong, taxon-specific anti-microbial effects as indicated by significant reductions in cell density (40%) and intra-sample community diversity. Members of the Bacteroidota were nearly eliminated over the 48-hour incubation. While generally part of a healthy gut microbiome, specific species of Bacteroides can be pathogenic. The active persistence of the members of the Enterobacteriaceae and selected Actinomycetota despite the reduction in overall cell numbers was demonstrated by increased fermentative outputs including high concentrations of gas and acetate with correspondingly reduced pH. These large-scale shifts in microbial community structure indicate the need for further evaluation of dosages and potential administration with prebiotic or synbiotic supplements. Overall, the very specific effects of these extracts may offer the potential for targeted antimicrobial uses or as a tool in the targeted remodeling of the gut microbiome. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a neurodegenerative disease that affects a wide range of populations and is the primary cause of death in various countries. The treatment of AD is still restricted to oral conventional medicines that act only superficially. Fabrication of intranasal solid lipid nanoparticulate system for the uptake of therapeutic agents will act as a convincing approach with limited off-site toxicity and increased pharmacological activity. The objective of this study was to formulate, optimize, and evaluate the efficiency of rivastigmine tartrate (RT)-loaded intranasal solid lipid nanoparticles (SLNs) employing the solvent-evaporation diffusion method. To optimize the formulation parameters, the central composite design (CCD) was used. Lipid concentration (X1) and surfactant concentration (X2) were considered to be independent variables, while particle size (Y1), percentage entrapment efficiency (Y2), and percentage drug release (Y3) were considered as responses. The solid lipid was glyceryl monostearate, while the surfactant was polysorbate 80. The optimized formulation has a particle size of 110.2 nm, % entrapment efficiency of 82.56%, and % drug release of 94.86%. The incompatibility of drug excipients was established by differential scanning calorimetry (DSC) and Fourier-transform infrared spectroscopy (FTIR). Nasal histopathology tests on sheep mucosa revealed that the developed SLNs were safe to utilize for intranasal delivery with no toxicity. Ex vivo permeation investigations revealed that the flux and diffusion coefficients for RT solid lipid nanoparticles and RT solution were 3.378 g/cm² /h and 0.310-3 cm² /h, respectively. Stability studies demonstrated that the developed SLNs were stable when stored under various storage conditions. The viability and vitality of adopting a lipid particle delivery system for improved bioavailability via the intranasal route were also established in the in vivo pharmacokinetic investigations. According to the histopathological and pharmacokinetic investigations, the developed formulations were safe, non-lethal, efficient, and robust. These results suggest the potentiality provided by rivastigmine tartrate-loaded solid lipid nanoparticles for nasal delivery. [ABSTRACT FROM AUTHOR]

Highly pathogenic avian influenza (HPAI) H5-viruses are circulating in wild birds and are repeatedly introduced to poultry causing outbreaks in the Netherlands since 2014. The largest epizootic ever recorded in Europe was caused by HPAI H5N1 clade 2.3.4.4b viruses in the period 2021–2022. The recent H5-clade 2.3.4.4 viruses were found to differ in their virulence for chickens and ducks. Viruses causing only mild disease may remain undetected, increasing the risk of virus spread to other farms, wild birds and mammals. We developed in ovo models to determine the virulence of HPAI viruses for chickens and ducks, which are fast and have low costs. The virulence of five contemporary H5-viruses was compared studying replication rate, average time to death and virus spread in the embryo. Remarkable differences in virulence were observed between H5-viruses and between poultry species. The H5N1-2021 virus was found to have a fast replication rate in both the chicken and duck in ovo models, but a slower systemic virus dissemination compared to three other H5-clade 2.3.4.4b viruses. The results show the potential of in ovo models to quickly determine the virulence of novel HPAI viruses, and study potential virulence factors which can help to better guide the surveillance in poultry. [ABSTRACT FROM AUTHOR]

Ex-vivo lung perfusion (EVLP) has extended the number of transplantable lungs by reconditioning marginal organs. However, EVLP is performed at 37°C without homeostatic regulation leading to metabolic wastes' accumulation in the perfusate and, as a corrective measure, the costly perfusate is repeatedly replaced during the standard of care procedure. As an interesting alternative, a hemodialyzer could be placed on the EVLP circuit, which was previously shown to rebalance the perfusate composition and to maintain lung function and viability without appearing to impact the global gene expression in the lung. Here, we assessed the biological effects of a hemodialyzer during EVLP by performing biochemical and refined functional genomic analyses over a 12h procedure in a pig model. We found that dialysis stabilized electrolytic and metabolic parameters of the perfusate but enhanced the gene expression and protein accumulation of several inflammatory cytokines and promoted a genomic profile predicting higher endothelial activation already at 6h and higher immune cytokine signaling at 12h. Therefore, epuration of EVLP with a dialyzer, while correcting features of the perfusate composition and maintaining the respiratory function, promotes inflammatory responses in the tissue. This finding suggests that modifying the metabolite composition of the perfusate by dialysis during EVLP can have detrimental effects on the tissue response and that this strategy should not be transferred as such to the clinic. [ABSTRACT FROM AUTHOR]

For recent decades, cardiac diseases have been the leading cause of death and morbidity worldwide. Despite significant achievements in their management, profound understanding of disease progression is limited. The lack of biologically relevant and robust preclinical disease models that truly grasp the molecular underpinnings of cardiac disease and its pathophysiology attributes to this stagnation, as well as the insufficiency of platforms that effectively explore novel therapeutic avenues. The area of fundamental and translational cardiac research has therefore gained wide interest of scientists in the clinical field, while the landscape has rapidly evolved towards an elaborate array of research modalities, characterized by diverse and distinctive traits. As a consequence, current literature lacks an intelligible and complete overview aimed at clinical scientists that focuses on selecting the optimal platform for translational research questions. In this review, we present an elaborate overview of current in vitro, ex vivo, in vivo and in silico platforms that model cardiac health and disease, delineating their main benefits and drawbacks, innovative prospects, and foremost fields of application in the scope of clinical research incentives. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis (Mtb) is the pathogen that causes tuberculosis and develops resistance to many of the existing drugs. The sole licensed TB vaccine, BCG, is unable to provide a comprehensive defense. So, it is crucial to maintain the immunological response to eliminate tuberculosis. Our previous in silico study reported five uncharacterized proteins as potential vaccine antigens. In this article, we considered the uncharacterized Mtb H37Rv regions of difference (RD-2) Rv1987 protein as a promising vaccine candidate. The vaccine quality of the protein was analyzed using reverse vaccinology and immunoinformatics-based quality-checking parameters followed by an ex vivo preliminary investigation. In silico analysis of Rv1987 protein predicted it as surface localized, secretory, single helix, antigenic, non-allergenic, and non-homologous to the host protein. Immunoinformatics analysis of Rv1987 by CD4 + and CD8 + T-cells via MHC-I and MHC-II binding affinity and presence of B-cell epitope predicted its immunogenicity. The docked complex analysis of the 3D model structure of the protein with immune cell receptor TLR-4 revealed the protein's capability for potential interaction. Furthermore, the target protein-encoded gene Rv1987 was cloned, over-expressed, purified, and analyzed by mass spectrometry (MS) to report the target peptides. The qRT-PCR gene expression analysis shows that it is capable of activating macrophages and significantly increasing the production of a number of key cytokines (TNF-α, IL-1β, and IL-10). Our in-silico analysis and ex vivo preliminary investigations revealed the immunogenic potential of the target protein. These findings suggest that the Rv1987 be undertaken as a potent subunit vaccine antigen and that further animal model immuno-modulation studies would boost the novel TB vaccine discovery and/or BCG vaccine supplement pipeline. [ABSTRACT FROM AUTHOR]

Sustainable livestock production requires reducing competition for food and feed resources and increasing the utilization of food by-products in livestock feed. This study describes the establishment of an anaerobic batch culture model to simulate pig microbiota and evaluate the effects of a food by-product, wakame seaweed stalks, on ex vivo microbial communities. We selected one of the nine media to support the growth of a bacterial community most similar in composition and diversity to that observed in pig donor feces. Supplementation with wakame altered the microbial profile and short-chain fatty acid composition in the ex vivo model, and a similar trajectory was observed in the in vivo pig experimental validation. Notably, the presence of wakame increased the abundance of Lactobacillus species, which may have been due to cross-feeding with Bacteroides. These results suggest the potential of wakame as a livestock feed capable of modulating the pig microbiome. Collectively, this study highlights the ability to estimate the microbiome changes that occur when pigs are fed a specific feed using an ex vivo culture model. [ABSTRACT FROM AUTHOR]

Accurate lymph node (LN) retrieval during colorectal carcinoma resection is pivotal for precise N-staging and the determination of adjuvant therapy. Current guidelines recommend the examination of at least 12 mesocolic or mesorectal lymph nodes for accurate staging. Traditional histological processing techniques, reliant on visual inspection and palpation, are time-consuming and heavily dependent on the examiner’s expertise and availability. Various methods have been documented to enhance LN retrieval from colorectal specimens, including intra-arterial ex vivo methylene blue injection. Recent studies have explored the utility of indocyanine green (ICG) fluorescence imaging for visualizing pericolic lymph nodes and identifying sentinel lymph nodes in colorectal malignancies. This study included 10 patients who underwent colon resection for malignant tumors. During surgery, intravenous ICG dye and an endoscopic camera were employed to assess intestinal perfusion. Post-resection, ex vivo intra-arterial administration of ICG dye was performed on the specimens, followed by routine histological processing and an ICG-assisted lymph node dissection. The objective was to evaluate whether ICG imaging could identify additional lymph nodes compared to routine manual dissection and to assess the clinical relevance of these findings. For each patient, a minimum of 12 lymph nodes (median = 25.5, interquartile range = 12.25, maximum = 33) were examined. ICG imaging facilitated the detection of a median of three additional lymph nodes not identified during routine processing. Metastatic lymph nodes were found in four patients however no additional metastatic nodes were detected with ICG assistance. Our findings suggest that ex vivo intra-arterial administration of indocyanine green dye can augment lymph node dissection, particularly in cases where the number of lymph nodes retrieved is below the recommended threshold of 12.

The development of drug therapy for the pathological calcification of the aortic valve is still an open issue due to the lack of effective treatment strategies. Currently, the only option for treating this condition is surgical correction and symptom management. The search for models to study the safety and efficacy of anti-calcifying drugs requires them to not only be as close as possible to in vivo conditions, but also to be flexible with regard to the molecular studies that can be applied to them. The ex vivo model has several advantages, including the ability to study the effect of a drug on human cells while preserving the original structure of the valve. This allows for a better understanding of how different cell types interact within the valve, including non-dividing cells. The aim of this study was to develop a reproducible ex vivo calcification model based on valves from patients with calcific aortic stenosis. We aimed to induce spontaneous calcification in valve tissue fragments under osteogenic conditions, and to demonstrate the possibility of significantly suppressing it using a calcification inhibitor. To validate the model, we tested a Notch inhibitor Crenigacestat (LY3039478), which has been previously shown to have an anti-calcifying effect on interstitial cell of the aortic valve. We demonstrate here an approach to testing calcification inhibitors using an ex vivo model of cultured human aortic valve tissue fragments. Thus, we propose that ex vivo models may warrant further investigation for their utility in studying aortic valve disease and performing pre-clinical assessment of drug efficacy.

Abstract Objectives To determine the imaging details and diagnostic information of the treatment response to neoadjuvant chemoradiotherapy (nCRT) of rectal adenocarcinoma at 9.4T magnetic resonance imaging (MRI) by ex vivo. Methods Fifteen cases with locally advanced rectal cancer (LARC) followed by radical surgery after nCRT between September 2022 and February 2023 were recruited. Resected specimens were fixed in a perfluoropolyether‐filled test tube and scanned with a 3.0T and 9.4T MRI system ex vivo. The residual tumor depth and MRI‐based tumor regression grade (TRG) were subjectively assessed and then compared with the pathological findings. Results The ex vivo 9.4T T2WI without fat suppression clearly differentiated tumor tissue, fibrosis and normal rectal wall, which clearly corresponded to the pathologic tissues of the rectal specimens. The TRG could be accurately assessed on ex vivo 9.4T images in 13/15 specimens (86.7%), while in 11/15 specimens (73.3%) on ex vivo 3.0T images. Conclusion Ex vivo 9.4T MR imaging clearly displayed the components of rectal wall and proved excellent diagnostic performance for evaluating the treatment response to nCRT, which allow radiologists to understand and then assess more accurately the TRG of LARC after nCRT.

Introduction: The development of biomaterials and medical devices for burn wound treatment necessitates thorough investigation through in vitro/ex vivo models before transitioning to animal studies. Establishing a standardized and high-throughput burn wound model in ex vivo skin presents a considerable challenge. Our objective was to address this challenge by developing a practical and cost-effective 3D-printed burn wound tool capable of uniformly inducing burns in 12 skin samples simultaneously. Material and methods: Utilizing Autodesk Inventor software, we designed a 3D model comprising a plate-base component (PBC) and a rod-base component (RBC). The design was exported as a Standard Triangulation Language (STL) file, processed through ''Slicer'' software to generate a G-code file tailored for 3D printing. Results: The Rod-Base component underwent iterative design modifications to optimize weight, airflow, and material consumption, resulting in a final design featuring a unique star shape for enhanced airflow. Simultaneously, the Plate-Base component design evolved to enable easy and secure plate placement, demonstrating compatibility with 12-well plates. The average production time for the model was 14.5 h, with a production cost of approximately $20 (USD), covering printing material and steel rods. Conclusion: In conclusion, this study provides valuable insights into the required equipment and software, empowering researchers to efficiently produce their accurate and cost-effective 3D-printed tool for controlled and reproducible burn wound creation in ex vivo viable skin tissues.

Lung diseases such as cancer substantially alter the mechanical properties of the organ with direct impact on the development, progression, diagnosis, and treatment response of diseases. Despite significant interest in the lung’s material properties, measuring the stiffness of intact lungs at sub-alveolar resolution has not been possible. Recently, we developed the crystal ribcage to image functioning lungs at optical resolution while controlling physiological parameters such as air pressure. Here, we introduce a data-driven, multiscale network model that takes images of the lung at different distending pressures, acquired via the crystal ribcage, and produces corresponding absolute stiffness maps. Following validation, we report absolute stiffness maps of the functioning lung at microscale resolution in health and disease. For representative images of a healthy lung and a lung with primary cancer, we find that while the lung exhibits significant stiffness heterogeneity at the microscale, primary tumors introduce even greater heterogeneity into the lung’s microenvironment. Additionally, we observe that while the healthy alveoli exhibit strain-stiffening of ∼1.75 times, the tumor’s stiffness increases by a factor of six across the range of measured transpulmonary pressures. While the tumor stiffness is 1.4 times the lung stiffness at a transpulmonary pressure of three cmH2O, the tumor’s mean stiffness is nearly five times greater than that of the surrounding tissue at a transpulmonary pressure of 18 cmH2O. Finally, we report that the variance in both strain and stiffness increases with transpulmonary pressure in both the healthy and cancerous lungs. Our new method allows quantitative assessment of disease-induced stiffness changes in the alveoli with implications for mechanotransduction.

Recently, the bioactive potential of several functional ingredients and biomolecules has been evaluated regarding human and animal nutrition. The digestive process from food intake to absorption and metabolism are important events that induce changes in ingredients, which affect their bioactivity. Consequently, there is a need to assess the bioavailability and bioaccessibility of these compounds. The methodology for the simulation of the human gastrointestinal tract has been standardized (INFOGEST protocol), while a gastrointestinal protocol for other animals (e.g., ruminants or broilers) has yet to be established. However, INFOGEST allows us only to predict bioaccessibility, leaving a gap regarding a methodology able to assess bioavailability by mimicking intestinal permeability and absorption. Several approaches—including in vitro, ex vivo, in situ and in vivo methods—can be found in the literature, aiming to tackle transepithelial routes, but leading to different results concerning the bioefficiency of the compounds studied. Therefore, this review aims to assess the current state-of-the-art regarding monogastric intestinal dynamics, absorption, and permeability events. Moreover, it compiled methodologies for simulating intestinal absorption in several biological systems, while reasoning their advantages, disadvantages, applications in ingredient development and the existing gaps.

Atrial fibrillation (AF) is the most common arrhythmia in the United States, affecting approximately 1 in 10 adults, and its prevalence is expected to rise as the population ages. Treatment options for AF are limited; moreover, the development of new treatments is hindered by limited (1) knowledge regarding human atrial electrophysiological endpoints (e.g., conduction velocity [CV]) and (2) accurate experimental models. Here, we measured the CV and refractory period, and subsequently calculated the conduction wavelength, in vivo (four subjects with AF and four controls), and ex vivo (atrial slices from human hearts). Then, we created an in vitro model of human atrial conduction using induced pluripotent stem (iPS) cells. This model consisted of iPS-derived human atrial cardiomyocytes plated onto a micropatterned linear 1D spiral design of Matrigel. The CV (34-41 cm/s) of the in vitro model was nearly five times faster than 2D controls (7-9 cm/s) and similar to in vivo (40-64 cm/s) and ex vivo (28-51 cm/s) measurements. Our iPS-derived in vitro model recapitulates key features of in vivo atrial conduction and may be a useful methodology to enhance our understanding of AF and model patient-specific disease.

Early gene therapy studies held great promise for the cure of heritable diseases, but the occurrence of various genotoxic events led to a pause in clinical trials and a more guarded approach to progress. Recent advances in genetic engineering technologies have reignited interest, leading to the approval of the first gene therapy product targeting genetic mutations in 2017. Gene therapy (GT) can be delivered either in vivo or ex vivo. An ex vivo approach to gene therapy is advantageous, as it allows for the characterization of the gene-modified cells and the selection of desired properties before patient administration. Autologous cells can also be used during this process which eliminates the possibility of immune rejection. This review highlights the various stages of ex vivo gene therapy, current research developments that have increased the efficiency and safety of this process, and a comprehensive summary of Human Immunodeficiency Virus (HIV) gene therapy studies, the majority of which have employed the ex vivo approach. [ABSTRACT FROM AUTHOR]

Currently, there are no ex vivo systems that can model the motion of peripheral arteries and allow for the evaluation of pharmacokinetics (PK) of endovascular devices. The objective of this study was to develop a novel peripheral simulating bioreactor system to evaluate drug pharmacokinetics of stents. We utilized 3D-printed and off-the-shelf components to construct a peripheral-simulating bioreactor system capable of mimicking the motion of peripheral arteries. Servo motors were primarily used to shorten/elongate, twist, and bend explanted porcine carotid arteries. To evaluate the pharmacokinetics in the bioreactor, drug-eluting stents were deployed within explanted arteries and subjected to vascular motion along with pulsatile flow conditions. Following 30 min and 24 h, the arteries were removed, and paclitaxel levels were measured. Scanning electron microscopy was also performed to evaluate the stent surface. Arterial paclitaxel levels of the stent-treated arteries were found to be higher at 30 min than at 24 h following pulsatile and no vascular motion and even higher at 24 h following pulsatile flow and vascular motion. The residual drug on the stent significantly decreased from 30 min to 24 h. Scanning electron microscopy confirmed the loss of paclitaxel coating at 24 h and greater disturbance in stents under peripheral motion versus pulsatile only. This system represents the first ex vivo system to determine the PK of drug-eluting stents under physiological flow and vascular motion conditions. This work provides a novel system for a quick and inexpensive preclinical tool to study acute drug tissue concentration kinetics of drug-releasing interventional vascular devices designed for peripheral applications. [ABSTRACT FROM AUTHOR]