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This study evaluated an alternative heating-stirring system in the determination of in vitro ruminal digestibility of dry matter (IVRDMD) of forages using the Tilley and Terry (TT) method. For this purpose, the IVRDMD of three forage species (Marandu, Tifton 85, and Mombasa) was determined by incubating 500 mg of each dried and ground (1 mm) forage in 50-mL rumen inoculum during 48h, followed by quantification of the incubation residue. Two heating-stirring systems were used: i) heating in a water bath at 39°C with manual stirring every two hours (i.e., traditional system); and ii) heating in an oven with controlled temperature at 39°C and automatic agitation (44 rpm; alternative system); there was no effect of the interaction between the heating-stirring system and the type of forage (p = 0.829) on the IVRDMD of forages. The type of heating-stirring system (p = 0.422) did not affect the IVRDMD of forages. Nevertheless, the IVRDMD values of Marandu grass (system i = 598.7 g kg-1 vs system ii = 599.4 g kg-1) were greater (p < 0.001) than Tifton 85 (system i = 392.1 vs g kg-1 vs system ii = 370.7 g kg-1) and Mombasa (system i = 397.4 g kg-1; system ii = 369.7 g kg-1) grasses. In conclusion, the obtained data indicate that the alternative heating-stirring system produces similar results to those obtained using the traditionally heating-stirring system during the determination of the IVRDMD of forages.

Information from the brain travels back and forth along peripheral nerves in the form of electrical impulses generated by neurons and these impulses have repetitive patterns. Schwann cells in peripheral nerves receive molecular signals from axons to coordinate the process of myelination. There is evidence, however, that non-molecular signals play an important role in myelination in the form of patterned electrical impulses generated by neuronal activity. The role of patterned electrical impulses has been investigated in the literature using co-cultures of neurons and myelinating cells. The co-culturing method, however, prevents the uncoupling of the direct effect of patterned electrical impulses on myelinating cells from the indirect effect mediated by neurons. To uncouple these effects and focus on the direct response of Schwann cells, we developed an in vitro model where an electroconductive carbon fiber acts as an artificial axon. The fiber provides only the biophysical characteristics of an axon but does not contribute any molecular signaling. In our \"suspended wire model\", the carbon fiber is suspended in a liquid media supported by a 3D printed scaffold. Patterned electrical impulses are generated by an Arduino 101 microcontroller. In this study, we describe the technology needed to set-up and eventually replicate this model. We also report on our initial in vitro tests where we were able to document the adherence and ensheath of human Schwann cells to the carbon fiber in the presence of patterned electrical impulses (hSCs were purchased from ScienCell Research Laboratories, Carlsbad, CA, USA; ScienCell fulfills the ethic requirements, including donor's consent). This technology will likely make feasible to investigate the response of Schwann cells to patterned electrical impulses in the future.

Respiratory infections, including tuberculosis, constitute a major global health challenge. Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains one of the leading causes of mortality worldwide. The disease’s complexity is attributed to Mtb’s capacity to persist in latent states, evade host immune defenses, and develop resistance to antimicrobial treatments, posing significant challenges for diagnosis and therapy. Traditional models, such as animal studies and two-dimensional (2D) in vitro systems, often fail to accurately recapitulate human-specific immune processes, particularly the formation of granulomas—a defining feature of tubercular infection. These limitations underscore the need for more physiologically relevant models to study TB pathogenesis. Emerging three-dimensional (3D) in vitro systems, including organoids and lung-on-chip platforms, offer innovative approaches to mimic the structural and functional complexity of the human lung. These models enable the recreation of key aspects of the tubercular granulomas, such as cellular interactions, oxygen gradients, and nutrient limitations, thereby providing deeper insights into Mtb pathogenesis. This review aims to elucidate the advantages of 3D in vitro systems in bridging the translational gap between traditional experimental approaches and clinical applications. Particular emphasis is placed on their potential to address challenges related to genetic variability in both the host and pathogen, thereby advancing tubercular research and therapeutic development.

Background Exposure to fine and ultra-fine ambient particles is still a problem of concern in many industrialised parts of the world and the intensified use of nanotechnology may further increase exposure to small particles. Complex in vitro coculture systems may be valuable tools to study particle-induced processes and to extrapolate effects of particles on the lung. A system consisting of four different human cell lines which mimics the cell response of the alveolar surface in vitro was developed to study native aerosol exposure (Vitrocell™ chamber). The system is composed of an alveolar type-II cell line (A549), differentiated macrophage-like cells (THP-1), mast cells (HMC-1) and endothelial cells (EA.hy 926), seeded in a 3D-orientation on a microporous membrane. Results The spatial distribution of the cells in the tetraculture was analysed by confocal laser scanning microscopy (CLSM), showing a confluent layer of endothelial and epithelial cells on both sides of the transwell. Macrophage-like cells and mast cells can be found on top of the epithelial cells. The cells formed colonies under submerged conditions, which disappeared at the ALI. To evaluate the response to oxidative stress, the dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used together with 2,2’-azobis-2-methyl-propanimidamide-dihydrochloride (AAPH) as inducer of oxidative stress. The tetraculture showed less induction of reactive oxygen species (ROS) production after being treated with a positive control compared to the monocultures of EA.hy 926, THP-1 and HMC-1. Submerged cultures showed elevated ROS and IL-8 levels compared to ALI cultures. The Vitrocell™ aerosol exposure system was not significantly influencing the viability. Using this system, cells were exposed to an aerosol of 50 nm SiO 2 -Rhodamine NPs in PBS. The distribution of the NPs in the tetraculture after exposure was evaluated by CLSM. Fluorescence from internalized particles was detected in CD11b-positive THP-1 cells only. Conclusion The system can be used in conjunction with a native aerosol exposure system and may finally lead to a more realistic judgement regarding the hazard of new compounds and/or new nano-scaled materials in the future. The results for the ROS production and IL-8 secretion suggest that submerged exposure may lead to an overestimation of observed effects.

Medicinal plants are used by many of the global population because they are safe and effective alternative to other treatments. Over the last few decades, using plant cells to produce natural or recombinant chemicals of economic interest has become a great intention. Secondary metabolites are recognized to have a significant role in plant adaptation to their environment and are also a valuable source of medication. The increasing economic importance of secondary metabolites has recently sparked a lot of interest in biotransformation, particularly in using tissue culture techniques to modify the production of bioactive plant metabolites. Plants of the family Amaryllidaceae have been used to extract several different alkaloids, each of which has the potential to be involved in a variety of pharmacological processes. Due to its multiple biological functions and divergent structure, lycorine has received significant interest in the medicinal field. Lycorine and other alkaloids from the Amaryllidaceae family have limited bioavailability by nature. In vitro culture provides an alternate method for producing lycorine sustainably due to the pharmaceutical industries dramatically increasing demand for it and the insufficient availability of natural resources. Many medicinal plants have been reported to produce lycorine in vitro in plant cell suspension cultures, and bioreactors play an effective role in their commercial production. This article focuses on the production of lycorine in in vitro systems from plants and its potential in the treatment of cancer. This study  also aims  to provide different biotechnological strategies  for the  production of this important alkaloid using in vitro system.

Tissue engineering is a multidisciplinary approach focused on the development of innovative bioartificial substitutes for damaged organs and tissues. For skeletal muscle, the measurement of contractile capability represents a crucial aspect for tissue replacement, drug screening and personalized medicine. To date, the measurement of engineered muscle tissues is rather invasive and not continuous. In this context, we proposed an innovative sensor for the continuous monitoring of engineered-muscle-tissue contractility through an embedded technique. The sensor is based on the calibrated deflection of one of the engineered tissue’s supporting pins, whose movements are measured using a noninvasive optical method. The sensor was calibrated to return force values through the use of a step linear motor and a micro-force transducer. Experimental results showed that the embedded sensor did not alter the correct maturation of the engineered muscle tissue. Finally, as proof of concept, we demonstrated the ability of the sensor to capture alterations in the force contractility of the engineered muscle tissues subjected to serum deprivation.

Respiratory sensitization as a consequence of exposure to chemical products has increased over the last decades, leading to an increase of morbidity. The increased use of synthetic compounds resulted in an exponential growth of substances to which we are potentially exposed on a daily basis. Some of them are known to induce respiratory sensitization, meaning that they can trigger the development of allergies. In the past, animal studies provided useful results for the understanding of mechanisms involved in the development of respiratory allergies. However, the mechanistic understanding of the involved cellular effects is still limited. Currently, no in vitro or in vivo models are validated to identify chemical respiratory sensitizers. Nonetheless, chemical respiratory sensitizers elicit a positive response in validated assays for skin sensitization. In this review, we will discuss how these assays could be used for respiratory sensitization and if necessary, what can be learnt from these assays to develop a model to assess the respiratory sensitizing potential of chemicals. In the last decades, much work has been done to study the respiratory toxicity of inhaled compounds especially in developing in vitro assays grown at the air–liquid interface. We will discuss how possibly the tests currently used to investigate general particle toxicity could be transformed to investigate respiratory sensitization. In the present review, we describe the most known mechanism involved in the sensitization process and the experimental in vivo and alternative in vitro models, which are currently available and how to adapt and improve existing models to study respiratory sensitization.

Anemia is a condition in which red blood cells or the hemoglobin in the blood is lower than in healthy people. Red blood cells transport and supply oxygen needed to various organs in the human body. Anemia is caused by hypoxemia due to the lack of red blood cells and causes other serious health problems, such as heart problems, pregnancy complications, severe fatigue, or death. There are many causes of anemia, and it can be diagnosed by measuring hematocrit or hemoglobin levels in the blood. Even though there are various diagnostic devices on the market, these devices are inconvenient because their systems are bulky, heavy, expensive, or inaccurate. This study proposed a new anemia diagnostic system based on the impedance measurement of red blood cells. The proposed system consists of a test strip that collects a blood sample from the finger and a hemoglobin meter that measures the impedance of the blood and converts it into the concentration of hemoglobin. The proposed test strip that does not contain enzymes or reagents was designed in accordance with class 1 approval by the Food and Drug Administration (FDA). The hemoglobin meter was designed to include a hardware block, an algorithm block and a calibration block through empirical work. We also compared it to reference impedance to prove the accuracy of the hemoglobin meter. The experimental results with human blood indicated the superiority of the anemia diagnostic system. As a result, the overall standard deviation of impedance measurements was less than 1%, and the coefficient of variance of the proposed system was 1.7%, which was better than that of other commercial systems.

Medicinal plants are important for improving human health and represent an essential pool for the identification of novel pharmacological leads. Plant-derived biomolecules have historically proven their value as a source of therapeutic drugs and hold an important potential for the identification and characterization of novel drug leads. Many different alkaloids possessing a broad range of pharmacological activities have been isolated from plants belonging to the Amaryllidaceae family. Galanthamine (GAL) is a selective, reversible and an Amaryllidaceae-derived acetylcholinesterase inhibitor used for the treatment of Alzheimer’s disease (AD) and other neurological diseases. Naturally, the bioavailability of Amaryllidaceae alkaloids including GAL is low. Due to the significantly increased demand of GAL by the pharmaceutical industries and the inadequate availability of natural resources, in vitro culture offers an alternative approach for its sustainable production. Thus, different biotechnological tools can optimize the in vitro GAL biosynthesis for treating AD, such as manipulation of plant growth regulators, photoperiod, elicitors, and bioreactors systems, besides being an environmentally sustainable approach, which protects the native biodiversity in a circular bioeconomy context. In the present review, we highlight the biosynthesis of GAL by plant in vitro systems including its mode of action. This article should also provide a starting point in the scaling-up of the biotechnological production of this valuable alkaloid.Key messageGalanthamine biosynthesis by plant in vitro systems and its mode of action are reviewed to provide a starting point for scaling-up of the biotechnological production of this valuable medicinal alkaloid.