Bioengineering

Hepatic cancer is widely regarded as the leading cause of cancer-related mortality worldwide. Despite recent advances in treatment options, the prognosis of liver cancer remains poor. Therefore, there is an urgent need to develop more representative in vitro models of liver cancer for pathophysiology and drug screening studies. Fortunately, an exciting new development for generating liver models in recent years has been the advent of organoid technology. Organoid models hold huge potential as an in vitro research tool because they can recapitulate the spatial architecture of primary liver cancers and maintain the molecular and functional variations of the native tissue counterparts during long-term culture in vitro. This review provides a comprehensive overview and discussion of the establishment and application of liver organoid models in vitro. Bioengineering strategies used to construct organoid models are also discussed. In addition, the clinical potential and other relevant applications of liver organoid models in different functional states are explored. In the end, this review discusses current limitations and future prospects to encourage further development.

Los alimentos, la medicina o el cuidado del medio ambiente son elementos basicos de la vida diaria de los cuales depende nuestro bienestar. La Nanotecnologia contribuye brindando seguridad y comodidad. El desarrollo de los sensores biologicos o biosensores son una herramienta que se ofrece para ser usada en areas tan diversas como Biomedicina y diagnostico clinico, jardines, analisis veterinario, analisis y control de la fermentacion, analisis y produccion de bebidas y alimentos, analisis de virus y bacterias, analisis farmaceutico, control de los residuos industriales, control y monitoreo de la contaminacion, mineria, gases toxicos e industriales, y hasta aplicaciones militares. Hasta hace unos pocos anos se consideraba que un biosensor era cualquier sonda analizadora que introducida en un medio biologico diera una senal cuantificable. Recientemente, la Nanotecnologia ha descubierto la resonancia plasmonica superficial (SPR), un fenomeno que puede ser la base de un biosensor muy po...

Neuronal entropy changes are observed in the basal ganglia circuit in Parkinson's disease (PD). These changes are observed in both single unit recordings from globus pallidus (GP) neurons and in local field potential (LFP) recordings from the subthalamic nucleus (STN). These changes are hypothesized as representing changes in the information coding capacity of the network, with PD resulting in a reduction in the coding capacity of the basal ganglia network. Entropy changes in the LFP and in single unit recordings are investigated in a detailed physiological model of the cortico-basal ganglia network during STN deep brain stimulation (DBS). The model incorporates extracellular stimulation of STN afferent fibers, with both orthodromic and antidromic activation, and simulation of the LFP detected at a differential recording electrode. LFP sample entropy and beta-band oscillation power were found to be altered following the application of DBS. The firing pattern entropy of GP neurons in the network were observed to decrease during high frequency stimulation and increase during low frequency stimulation. Simulation results were consistent with experimentally reported changes in neuronal entropy during DBS.

Neuronal entropy changes are observed in the basal ganglia circuit in Parkinson's disease (PD). These changes are observed in both single unit recordings from globus pallidus (GP) neurons and in local field potential (LFP) recordings from the subthalamic nucleus (STN). These changes are hypothesized as representing changes in the information coding capacity of the network, with PD resulting in a reduction in the coding capacity of the basal ganglia network. Entropy changes in the LFP and in single unit recordings are investigated in a detailed physiological model of the cortico-basal ganglia network during STN deep brain stimulation (DBS). The model incorporates extracellular stimulation of STN afferent fibers, with both orthodromic and antidromic activation, and simulation of the LFP detected at a differential recording electrode. LFP sample entropy and beta-band oscillation power were found to be altered following the application of DBS. The firing pattern entropy of GP neurons in the network were observed to decrease during high frequency stimulation and increase during low frequency stimulation. Simulation results were consistent with experimentally reported changes in neuronal entropy during DBS.

This study presents a modelling framework in which information on muscle fiber direction and orientation during contraction is derived from diffusion tensor imaging (DTI) and incorporated in a computational model of the surface electromyographic (EMG) signal. The proposed model makes use of the principle of reciprocity to simultaneously calculate the electric potentials produced at the recording electrode by charges distributed along an arbitrary number of muscle fibers within the muscle, allowing for a computationally efficient evaluation of extracellular motor unit action potentials. The approach is applied to the complex architecture of the first dorsal interosseous (FDI) muscle of the hand to simulate EMG during index finger flexion and abduction. Using diffusion tensor imaging methods, the results show how muscle fiber orientation and curvature in this intrinsic hand muscle change during flexion and abduction. Incorporation of anatomically accurate muscle architecture and other hand tissue morphologies enables the model to capture variations in extracellular action potential waveform shape across the motor unit population and to predict experimentally observed differences in EMG signal features when switching from index finger abduction to flexion. The simulation results illustrate how structural and electrical properties of the tissues comprising the volume conductor, in combination with fiber direction and curvature, shape the detected action potentials. Using the model, the relative contribution of motor units of different sizes located throughout the muscle under both conditions is examined, yielding a prediction of the detection profile of the surface EMG electrode array over the muscle crosssection.

Fungi are among the most degradative organisms inducing biodeterioration of paper-based items of cultural heritage. Appropriate conservation measures and restoration treatments to deal with fungal infections include mechanical, chemical, and biological methods, which entail effects on the paper itself and health hazards for humans. Three different conservation treatments, namely freeze-drying, gamma rays, and ethylene oxide fumigation, were compared and monitored to assess their short-(one month, T1) and long-term (one year, T2) effectiveness to inhibit fungal growth. After the inoculation with fungi possessing cellulose hydrolysis ability d Chaetomium globosum, Trichoderma viride, and Cladosporium cladosporioides d as single strains or as a mixture, different quality paper samples were treated and screened for fungal viability by culture-dependent and -independent techniques. Results derived from both strategies were contradictory. Both gamma irradiation and EtO fumigation showed full efficacy as disinfecting agents when evaluated with cultivation techniques. However, when using molecular analyses, the application of gamma rays showed a short-term reduction in DNA recovery and DNA fragmentation; the latter phenomenon was also observed in a minor degree in samples treated with freeze-drying. When RNA was used as an indicator of long-term fungal viability, differences in the RNA recovery from samples treated with freeze-drying or gamma rays could be observed in samples inoculated with the mixed culture. Only the treatment with ethylene oxide proved negative for both DNA and RNA recovery. Therefore, DNA fragmentation after an ethylene oxide treatment can hamper future paleogenetic and archaeological molecular studies on the objects.

Silybum marianum (SM) has attracted substantial attention because it is been used as a gold medicinal herb in traditional folk medicine for treatment of liver diseases and it possesses other wondrous health benefits. To support its functional attributes, various investigations have been carried out to find out its antioxidant, anti-inflammatory, anti-antiviral, anti-diabetic, anti-amnesia, cardio-protection, hepatic protective activities, the efficacy in the treatment of obsessive-compulsive disorder, sepsis and burn prevent and application on veterinary under publications of certain types of articles including "original research and short communications". Some publications either reiterate previously reports or create slight mistakes in performing methods, the problem associated quality performance to claim exactly biological activities found in the literature. Furthermore, correct expressions and original citations of the relevant models are provided to interpret and clarify. The authors hope that this work will be helpful to give insight knowledge for readers, researchers, reviewers, and editors who are interested in the related field of the SM studies.

Phosphate acquisition by plants is an essential process that is directly implicated in the optimization of crop yields. Purple acid phosphatases (PAPs) are ubiquitous metalloenzymes, which catalyze the hydrolysis of a wide range of phosphate esters and anhydrides. While some plant PAPs display a preference for ATP as the substrate, others are efficient in hydrolyzing phytate or 2-phosphoenolpyruvate (PEP). PAP from red kidney bean (rkbPAP) is an efficient ATP-and ADPase, but has no activity towards phytate. The crystal structure of this enzyme in complex with an ATP analogue (to 2.20 Å resolution) provides insight into the amino acid residues that play an essential role in binding this substrate. Homology modelling was used to generate three-dimensional structures for the active sites of PAPs from tobacco (NtPAP) and Arabidopsis thaliana (AtPAP12 and AtPAP26) that are efficient in hydrolyzing phytate and PEP as substrates, respectively. In combination with substrate docking simulations and a phylogenetic analysis of 49 plant PAP sequences (including the first PAP sequences reported from Eucalyptus), several active site residues were identified that are important in defining the substrate specificities of plant PAPs. These results may inform bioengineering studies aimed at identifying and incorporating suitable plant PAP genes into crops to improve phosphorus use efficiency. Organic phosphorus sources increasingly supplement or replace inorganic fertilizer, and efficient phosphorus use of crops will lower the environmental footprint of agriculture while enhancing food production.

Cronobacter sakazakii (Enterobacter sakazakii) is an emerging pathogen that causes necrotizing enterocolitis, sepsis, and meningitis. This bacterium is opportunistic pathogen and is linked with life-threatening infections in neonates and elderly persons. It is considered as ubiquitous organism and can be found in a broad range of foods & food ingredients and in water, in a variety of areas, including hospitals and houses though outbreaks most commonly associated with the ingestion of contaminated powdered infant formula (PIF). International Commission on Microbiological Specifications for Foods has ranked C. sakazakii a "severe hazard for restricted populations." It poses high tolerance to environmental stresses such as osmotic stress, elevated temperature etc. and decontamination processes and has broad antibiotic resistance and resistance to bile salts and disinfectants. Cronobacter sakazakii may survive in macrophage cells and efficiently attach to and invade epithelial cell lines, produce exopolysaccharide, form biofilm and has active efflux pumps. Controlling the organism in the production environment, thereby reducing dissemination, necessitates the provision of suitable diagnostic tools. Appropriate measures by parents, food and infant formula manufacturers, and health care providers, as well as understanding of the pathogenesis, are important in the prevention of C. sakazakii-related infections.

University Linear programming models are widely used for farm-level investment decisions. The particular advantage of using this spatial decision support system is its ability to include region-wide competitive forces and local, national and international market constraints. The most apparent advantages of the optimisation technique can be summarised as follows: .:. The technique integrated resource potential and economic determinants in predicting land-use patterns. This interactive capability determined the relative profitability and competitive advantage of each of the selected crops vis-a-vis the resource units.

The biomass supply chain constitutes a system that is highly dynamic and stochastic. The developed and proposed systems architectures for the management of the supply chains of typical industrial products do not directly apply to the case of the biomass supply chain. Nevertheless, it is imperative that a dedicated framework is needed according to the industrial standards. In this paper, a framework for the biomass Supply Chain Event Management (SCEM) supporting the efficient management of inter-organizational events within the examined supply chain is proposed and analyzed. The conceptual model includes the identification and specification of events and the preparation of necessary notification based on information needs.

A reverse transcriptase-polymerase chain reaction was used to isolate a gene for βglucosidase from fungi Aspergillus terreus SUK-1. The DNA consisted of 2586 nucleotides coding sequence encoding 861 amino acids for BGat1 protein. Both nucleotide and amino acid sequences possessed high similarity match with reported β-glucosidase genes and proteins such as of Aspergillus spp. and Neosartorya fischeri. The presence of a 19 amino acids leader sequence suggested that BGat1 is an extracellular protein and very conserve with 5 other β-glucosidases tested. Two specific domains, namely GHF3 N terminal domain and GHF3 C terminal domain, were detected in BGat1 and these domains were separated by a spacer of 99 amino acids. This suggests that BGat1 is a member of glycosyl hydrolase superfamily 3 proteins. Expression analysis showed that BGat1 gene was induced as early as 7 hours after fungal incubation and the level of gene expression maintained at almost the same until day 7. BGat1 expression was also detected when A. terreus SUK-1 was grown in the presence of glucose suggesting that the gene was not inhibited by the product of enzymatic reaction.

This work aims to engineer a new stable injectable Mn-based methacrylated gellan gum (Mn/GG-MA) hydrogel for real-time monitored cell delivery into the central nervous system. To enable the hydrogel visualization under Magnetic Resonance Imaging (MRI), GG-MA solutions were supplemented with paramagnetic Mn2+ ions before its ionic crosslink with artificial cerebrospinal fluid (aCSF). The resulting formulations were stable, detectable by T1-weighted MRI scans and also injectable. Cell-laden hydrogels were prepared using the Mn/GG-MA formulations, extruded into aCSF for crosslink, and after 7 days of culture, the encapsulated human adipose-derived stem cells remained viable, as assessed by Live/Dead assay. In vivo tests, using double mutant MBPshi/shi/rag2 immunocompromised mice, showed that the injection of Mn/GG-MA solutions resulted in a continuous and traceable hydrogel, visible on MRI scans. Summing up, the developed formulations are suitable for both non-invasive cell delivery te...

Microemulsions and nanoemulsions are lipid-based pharmaceutical systems with a high potential to increase the permeation of drugs through the skin. Although being isotropic dispersions of two nonmiscible liquids (oil and water), significant differences are encountered between microemulsions and nanoemulsions. Microemulsions are thermodynamically stable o/w emulsions of mean droplet size approximately 100–400 nm, whereas nanoemulsions are thermodynamically unstable o/w emulsions of mean droplet size approximately 1 to 100 nm. Their inner oil phase allows the solubilization of lipophilic drugs, achieving high encapsulation rates, which are instrumental for drug delivery. In this review, the importance of these systems, the key differences regarding their composition and production processes are discussed. While most of the micro/nanoemulsions on the market are held by the cosmetic industry to enhance the activity of drugs used in skincare products, the development of novel pharmaceuti...

Surfactants are amphiphilic compounds having hydrophilic and hydrophobic moieties in their structure. They can be of synthetic or of microbial origin, obtained respectively from chemical synthesis or from microorganisms’ activity. A new generation of ecofriendly surfactant molecules or biobased surfactants is increasingly growing, attributed to their versatility of applications. Surfactants can be used as drug delivery systems for a range of molecules given their capacity to create micelles which can promote the encapsulation of bioactives of pharmaceutical interest; besides, these assemblies can also show antimicrobial properties. The advantages of biosurfactants include their high biodegradability profile, low risk of toxicity, production from renewable sources, functionality under extreme pH and temperature conditions, and long-term physicochemical stability. The application potential of these types of polymers is related to their properties enabling them to be processed by emuls...

Introduction Tissue engineered skin replacements have opened new options to treat skin loss. Although increased healing rates of burn and/or chronic wounds can be observed with those engineered constructs, several intrinsic shortcomings still limit their use, including inadequate adhesiveness and mechanical properties, and sub-optimal manufacturing processes (duration, costs). In this context, bioreactors could be the key to accomplish reliable, traceable and cost-effective tissue engineered processes, with minimal risks of infection. Moreover, stretching proved to be a key stimulus for cell growth, differentiation and migration, tissue remodeling and gene expression [1,2]. The aim of the present study is to investigate the role of mechanical conditioning in keratinocyte monolayer cultures using our previously developed bioreactor (Figure 1). Cell viability and morphology, and matrix metalloproteinases (MMP) 2, 9 and 28 – shown to be implicated in cell migration, proliferation and t...

The HIV/AIDS pandemic and its impact on women prompt the investigation of prevention strategies to interrupt sexual transmission of HIV. Long-acting drug delivery systems that simultaneously protect womenfrom sexual transmission of HIV and unwanted pregnancy could be important tools in combating the pandemic. We describe the design, in silico, in vitro and in vivo evaluation of a dual-reservoir intravaginal ring that delivers the HIV-1 reverse transcriptase inhibitor tenofovir and the contraceptive levonorgestrel for 90 days. Two polyether urethanes with two different hard segment volume fractions were used to make coaxial extruded reservoir segments with a 100 mm thick rate controlling membrane and a diameter of 5.5 mm that contain 1.3 wt% levonorgestrel. A new mechanistic diffusion model accurately described the levonorgestrel burst release in early time points and pseudo-steady state behavior at later time points. As previously described, tenofovir was formulated as a glycerol paste and filled into a hydrophilic polyurethane, hollow tube reservoir that was melt-sealed by induction welding. These tenofovir-eluting segments and 2 cm long coaxially extruded levonorgestrel eluting segments were joined by induction welding to form rings that released an average of 7.5 mg tenofovir and 21 mg levonorgestrel per day in vitro for 90 days. Levonorgestrel segments placed intravaginally in rabbits resulted in sustained, dose-dependent levels of levonorgestrel in plasma and cervical tissue for 90 days. Polyurethane caps placed between segments successfully prevented diffusion of levonorgestrel into the tenofovir-releasing segment during storage.Hydrated rings endured between 152 N and 354 N tensile load before failure during uniaxial extension testing. In summary, this system represents a significant advance in vaginal drug delivery technology, and is the first in a new class of long-acting multipurpose prevention drug delivery systems.

The field of dental medicine is constantly evolving and advancing toward minimally invasive techniques. Several studies have demonstrated that bonding to the tooth structure, particularly enamel, yields the most predictable results. In some instances, however, significant tooth loss, pulpal necrosis, or irreversible pulpitis may limit the options available to the restorative dentist. In these cases, placement of a post and core followed by a crown is the preferred treatment option, provided all requirements are met. This literature review provides an overview of the historical development of dental FRC post systems as well as a comprehensive examination of the currently available posts and their bonding requirements. In addition, it offers valuable insights for dental professionals seeking to understand the current state of the field and the prospects of dental FRC post systems.

Este artículo presenta un modelo circuital que puede ser empleado para evaluar las curvas obtenidas en la prueba de respuesta en frecuencia del transformador. El modelo es sensible a cambios en la curvas, por lo que puede ser de gran utilidad para determinar las posibles condiciones del transformador que sean las causas de las variaciones. Se hace la descripción del modelo y se muestran algunos casos de su aplicación.

Autologous bone replacement remains the preferred treatment for segmental defects of the mandible; however, it cannot replicate complex facial geometry and causes donor site morbidity. Bone tissue engineering has the potential to overcome these limitations. Various commercially available calcium phosphate-based bone substitutes (Novabone®, BioOss®, and Zengro®) are commonly used in dentistry for small bone defects around teeth and implants. However, their role in ectopic bone formation, which can later be applied as vascularized graft in a bone defect, is yet to be explored. Here, we compare the above-mentioned bone substitutes with autologous bone with the aim of selecting one for future studies of segmental mandibular repair. Six female sheep, aged 7–8 years, were implanted with 40 mm long four-chambered polyether ether ketone (PEEK) bioreactors prepared using additive manufacturing followed by plasma immersion ion implantation (PIII) to improve hydrophilicity and bioactivity. Eac...

Transplantation of human neural stem cells (hNSCs) is emerging as a viable treatment for stroke related brain injury. However, intraparenchymal grafts do not regenerate lost tissue, but rather integrate into the host parenchyma without significantly affecting the lesion cavity. Providing a structural support for the delivered cells appears important for cell based therapeutic approaches. The non-invasive monitoring of therapeutic methods would provide valuable information regarding therapeutic strategies but remains a challenge. Labeling transplanted cells with metalbased 1 H-magnetic resonance imaging (MRI) contrast agents affects the visualization of the lesion cavity. Herein, we demonstrate that a 19 F-MRI contrast agent can adequately monitor the

We report here the construction of a vector derived from pET3-His and pRSET plasmids for the expression and purification of recombinant proteins in Escherichia coli based on T7 phage RNA polymerase. The resulting pAE plasmid combined the advantages of both vectors: small size (pRSET), expression of a short 6XHis tag at N-terminus (pET3-His) and a high copy number of plasmid (pRSET). The small size of the vector (2.8 kb) and the high copy number/cell (200-250 copies) facilitate the subcloning and sequencing procedures when compared to the pET system (pET3-His, 4.6 kb and 40-50 copies) and also result in high level expression of recombinant proteins (20 mg purified protein/liter of culture). In addition, the vector pAE enables the expression of a fusion protein with a minimal amino-terminal hexahistidine affinity tag (a tag of 9 amino acids using XhoI restriction enzyme for the 5'cloning site) as in the case of pET3-His plasmid and in contrast to proteins expressed by pRSET plasmids (a tag of 36 amino acids using BamHI restriction enzyme for the 5'cloning site). Thus, although proteins expressed by pRSET plasmids also have a hexa-histidine tag, the fusion peptide is much longer and may represent a problem for some recombinant proteins.

Electrospinning is a promising technique for the production of scaffolds aimed at the regeneration of soft tissues. The aim of this work was to develop electrospun bundles mimicking the architecture and mechanical properties of the fascicles of the human Achille tendon. Two different blends of poly(L-lactic acid) (PLLA) and collagen (Coll) were tested, PLLA/Coll-75/25 and PLLA/Coll-50/50, and compared with bundles of pure PLLA. First, a complete physico-chemical characterization was performed on non-woven mats made of randomly arranged fibers. The presence of collagen in the fibers was assessed by thermogravimetric analysis, differential scanning calorimetry and water contact angle measurements. The collagen release in phosphate buffer solution (PBS) was evaluated for 14 days: results showed that collagen loss was about 50% for PLLA/Coll-75/25 and 70% for PLLA/Coll-50/50. In the bundles, the individual fibers had a diameter of 0.48± 0.14 µm (PLLA), 0.31±0.09 µm (PLLA/Coll-75/25), 0.33±0.08 µm (PLLA/Coll-50/50), whereas bundle diameter was in the range 300-500 µm for all samples. Monotonic tensile tests were performed to measure the mechanical properties of PLLA bundles (as-spun) and of PLLA/Coll-75/25 and PLLA/Coll-50/50 bundles (as-spun, and after 48 hour, 7 days and 14 days in PBS). The most promising material was the PLLA/Coll-75/25 blend with a Young modulus of 98.6±12.4 MPa (as-spun) and 205.1±73.0 MPa (after 14 days in PBS). Its failure stress was 14.2±0.7 MPa (as-spun) and 6.8±0.6 MPa (after 14 days in PBS). Pure PLLA withstood slightly lower stress than the PLLA/Coll-75/25 while PLLA/Coll-50/50 had a brittle behavior. Humanderived tenocytes were used for cellular tests. A good cell adhesion and viability after 14 day culture was observed. This study has therefore demonstrated the feasibility of fabricating electrospun bundles with multiscale structure and mechanical properties similar to the human tendon.

Articular cartilage is a tissue of fundamental importance for the mechanics of joints, since it provides a smooth and lubricated surface for the proper transfer of loads. From a mechanical point of view, this tissue is an anisotropic poroviscoelastic material: its characteristics at the macroscopic level depend on the complex microscopic architecture. With the ability to probe the local microscopic features, dynamic nanoindentation test is a powerful tool to investigate cartilage mechanics. In this work we focus on a length scale where the time dependent behaviour is regulated by poroelasticity more than viscoelasticity and we aim to understand the effect of the anisotropic permeability on the mechanics of the superficial layer of the articular cartilage. In a previous work, a finite element model for the dynamic nanoindentation test has been presented. In this work, we improve the model by considering the presence of an anisotropic permeability tensor that depends on the collagen fibers distribution. Our sensitivity analysis highlights that the permeability decreases with increasing indentation, thus making the tissue stiffer than the case of isotropic permeability, when solicited at the same frequency. With this improved model, a revised identification of the mechanical and physical parameters for articular cartilage is provided. To this purpose the model was used to simulate experimental data from tests performed on bovine tissue, giving a better estimation of the anisotropy in the elastic properties. A relation between the identified macroscopic anisotropic permeability properties and the microscopic rearrangement of the fiber/matrix structure during indentation is also provided.

The larvae of scarab beetles, known as “white grubs” and belonging to the genera Phyllophaga and Anomala (Coleoptera: Scarabaeidae), are regarded as soil-dwelling pests in Mexico. During a survey conducted to find pathogenic bacteria with the potential to control scarab larvae, a native Serratia sp. (strain Mor4.1) was isolated from a dead third-instar Phyllophaga blanchardi larva collected from a cornfield in Tres Marías, Morelos, Mexico. Oral bioassays using healthy P. blanchardi larvae fed with the Mor4.1 isolate showed that this strain was able to cause an antifeeding effect and a significant loss of weight. Mortality was observed for P. blanchardi , P. trichodes , and P. obsoleta in a multidose experiment. The Mor4.1 isolate also caused 100% mortality 24 h after intracoelomic inoculation of the larvae of P. blanchardi , P. ravida , Anomala donovani and the lepidopteran insect Manduca sexta . Oral and injection bioassays were performed with concentrated culture broths of the Mo...

Atactic poly(N-isopropyl acrylamide) (PNIPAM) is a thermosensitive polymer in aqueous solutions. By modifying its structure with isotactic (i) rich blocks it has been shown that the thermal and structural properties of the solutions can be modified. For example, a triblock PNIPAM with isotactic endblocks forms hydrogels at temperatures close to room temperature. These gels phase separate at higher temperatures and dissolve when lowering the temperature. The rheological properties of PNIPAM polymers in water-acetone mixtures were investigated to see how the acetone content affects solution behaviour. Different arrangements of the atactic and isotactic block result in different complex viscosity as a function of temperature profiles and the block arrangement have an effect on the strength of the formed gel. It was also found that the acetone content influences the gelling behaviour of the PNIPAM systems to a large extent.

Detection and identification of bovine tuberculosis and its differentiation from micobacteriosis is fundamental during diagnoses. That is why mycobacteria laboratories improvement becomes essential in public health and veterinary medicine services. The objective of the present research is to differentiate Mycobacterium bovis and nontuberculous mycobacterias in isolates cultured from domestic and wildlife species from seven Argentinean provinces during [2004][2005][2006][2007][2008]. Differentiation was based on biochemical tests, phenotypic characteristics and M. bovis spolygotyping. Biochemical and phenotypic identification resulted in 20 M. bovis strains, 18 of them were confirmed by spolygotyping, and 34 nontuberculous mycobacteria strains. Thirteen species were characterized and all of them were grouped considering biological risk and pathogenic potential reported in humans and/or animals. Here we have reached advances in tuberculosis and micobacteriosis diagnoses in veterinary medicine. In this area diagnoses are often based on micro and macroscopic observation of the tubercles and skin test results. These advances are not minor as zoonotic tuberculosis is still a public health problem in Latin America.

This research is engaged to explore biological peristaltic transport under the action of an externally applied magnetic field passing through an asymmetric channel which is saturated with porous media. The set of governing partial differential equations for the present peristaltic flow are solved in the absence of a low Reynolds number and long wavelength assumptions. The governing equations are to be solved completely, so that inertial effects can be studied. The numerical simulations and results are obtained by the help of a finite element method based on quadratic six-noded triangular elements equipped with a Galerkin residual procedure. The inertial effects and effects of other pertinent parameters are discussed by plotting graphs based on a finite element (FEM) solution. Trapped bolus is discussed using the graphs of streamlines. The obtained results are also compared with the results given in the literature which are highly convergent. It is concluded that velocity and the num...

The application of nanoparticles has raised concern over the safety of these materials to human health and the ecosystem. After release into an aquatic environment, nanoparticles are likely to experience heteroaggregation with biocolloids, geocolloids, natural organic matter (NOM) and other types of nanoparticles. Heteroaggregation is of vital importance for determining the fate and transport of nanoparticles in aqueous phase and sediments. In this article, we review the typical cases of heteroaggregation between nanoparticles and biocolloids and/or geocolloids, mechanisms, modeling, and important indicators used to determine heteroaggregation in aqueous phase. The major mechanisms of heteroaggregation include electric force, bridging, hydrogen bonding, and chemical bonding. The modeling of heteroaggregation typically considers DLVO, X-DLVO, and fractal dimension. The major indicators for studying heteroaggregation of nanoparticles include surface charge measurements, size measureme...

• Iron-based coagulants (PFS and FeCl 3 ) are effective to remove TiO 2 nanoparticles. • Water chemistry influences TiO 2 nanoparticles removal. • The effective pH range for PFS is broader than for FeCl 3 . • Humic acid stabilizes TiO 2 nanopar- ticles and reduces removal by steric hindrance.

The main purpose of this research is to improve the production of crude triterpenoid and extracellular polysaccharide of Lignosus rhinocerus by liquid fermentation to satisfy market demand. As we all known, the low yield of the two bioactive metabolites is a difficult problem for further research. In this paper, eleven aqueous herbal extracts were added to the fermentation of L. rhinocerus to increase the final concentration of the two target products. According to the ‘one-factor-at-a-time’ method, Radix polygoni multiflora and Chamomile extract were identified to be effective elicitors for the improvement of mycelial biomass, cEPS and crude triterpenoid production. The peak values of mycelial biomass (i.e., 23.97 ± 0.31 g/L) and cEPS (i.e., 6.97 ± 0.56 g/L) were obtained under the concentration of 15 and 5 g/L from Radix polygoni multiflora extract, respectively. Moreover, the maximum crude triterpenoid production reached 8.72 ± 0.20 mg/g with the addition of Chamomile extract und...

The increasing commercial and biotechnological interest in yeast cell wall components has brought about the search of low cost sources. One potential source may be the yeast cultures obtained from fermentation processes and particularly, those that are usually discarded afterwards. In this contribution, we isolated five yeast strains from the agave must and found, next to Saccharomyces cerevisiae (CTGM, CTSA) and Meyerozyma guilliermondii (CT15, CT25, CT35), which has not been reported to be involved in the tequila production before. All strains were grown non shaken in agave must with 30 gL-1 of sugars and 5 gL-1 of (NH4)2SO4. It was found that the cell wall composition was more variable among the strains than between the species and highly depended on the growth phases. In the stationary phase, only two strains (CT15 and CT25) had a high b-glucan content in the cell wall (~30 mg g-1) while the others yielded only 3 to 18 mg g-1, also the control bakery yeast strain. This indicates...

Taxus wallichiana Zucc., is a medium sized temperate forest tree species of Asia ranging from Afghanistan through the Himalayas to the Philippines. It has been heavily exploited for its leaves and bark which are used to produce the anti-cancer drug Taxol. Due to its long seed dormancy period, its natural regeneration from seeds is very low. In the present study, biotechnological method is applied to grow the plant In vitro through plant tissue culture techniques. Different plant growth regulators (2, 4-D, NAA, IBA, BAP, Kin) were used to regenerate it through direct or indirect route of organogenesis in the In vitro conditions. Callus was induced successfully both in stem and leaf (needle) explant on MS media supplemented with 1 mg/L, 1.5 mg/L, 2 mg/L, 2.5 mg/L and 3.0 mg/L, 2,4-D. The best callus was induced on MS media supplemented with 2 mg/L 2,4-D and 5 mg/L Activated Charcoal (AC) within 2 weeks of culture in stem explant, but we did not succeeded in the regenerations of shoot in both type of callus culture. The shoot tip meristem was elongated on MS media supplemented with 2 mg/L BAP and MS media supplemented with 1mg/L IBA up to 10-14 cm after 3 to 4 subculture. Roots were induced in the elongated shoot tips in 60-80 days on MS media supplemented with 3.5 mg/L IBA and on half strength MS media supplemented with 8 mg/L IBA. It is concluded from the present study that callus culture from stem and needle explant is not suitable for organogenesis in Taxus wallichiana, however shoot elongation and root induction in shoot tip culture is feasible and suitable for the multiplication of Taxus wallichiana through tissue culture.

In order efficiently to target therapies intending to stop or reverse degenerative processes of articular cartilage, it would be crucial to diagnose osteoarthritis (OA) earlier and more sensitively than is possible with the existing clinical methods. Unfortunately, current clinical methods for OA diagnostics are insensitive for detecting the early degenerative changes, e.g., arising from collagen network damage or proteoglycan depletion. We have recently investigated several novel quantitative biophysical methods, including ultrasound indentation, quantitative ultrasound techniques and magnetic resonance imaging, for diagnosing the degenerative changes of articular cartilage, typical for OA. In this study, the combined results of these novel diagnostic methods were compared with histological (Mankin score, MS), compositional (proteoglycan, collagen and water content) and mechanical (dynamic and equilibrium moduli) reference measurements of the same bovine cartilage samples. Receiver operating characteristics (ROC) analysis was conducted to judge the diagnostic performance of each technique. Indentation and ultrasound techniques provided the most sensitive measures to differentiate samples of intact appearance (MS= 0) from early (1<MS<3) or more advanced (MS>3) degeneration. Furthermore, these techniques were good predictors of tissue composition and mechanical properties. The specificity and sensitivity analyses revealed that the mechano-acoustic methods, when further developed for in vivo use, may provide more sensitive probes for OA diagnostics than the prevailing qualitative X-ray and arthroscopic techniques. Noninvasive quantitative MRI measurements showed slightly lower diagnostic performance than mechano-acoustic techniques. The compared methods could possibly also be used for the quantitative monitoring of success of cartilage repair.

Acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum has been extensively studied in recent years because the organism is recognized as an excellent butanol producer. A parallel bioreactor system with 48 stirred-tank bioreactors on a 12 mL scale was evaluated for batch cultivations of the strictly anaerobic, butanol-producing C. acetobutylicum ATCC 824. Continuous gassing with nitrogen gas was applied to control anaerobic conditions. Process performances of ABE batch fermentations on a milliliter scale were identical to the liter-scale stirred-tank reactor if reaction conditions were identical on the different scales (e.g., initial medium, pH, temperature, specific evaporation rates, specific power input by the stirrers). The effects of varying initial ammonia concentrations (0.1 -4.4 g L -1 ) were studied in parallel with respect to glucose consumption and butanol production of C. acetogbutylicum ATCC 824 as a first application example. The highest butanol yield of 33 % (mol mol -1 ) was observed at initial ammonia concentrations of 0.5 and 1.1 g L -1 . This is the first report on successful application of a 48 parallel stirred-tank bioreactor system for reaction engineering studies of strictly anaerobic microorganisms at milliliter scale.

Targeting the spleen with nanoparticles could increase the efficacy of vaccines and cancer immunotherapy, and have the potential to treat intracellular infections including leishmaniasis, trypanosome, splenic TB, AIDS, malaria, and hematological disorders. Although, nanoparticle capture in both the liver and spleen has been well documented, there are only a few examples of specific capture in the spleen alone. It is proposed that the larger the nanoparticle size (> 400 nm) the greater the specificity and capture within the spleen. Here, we synthesized five nanostructures with different shapes (ranging from spheres, worms, rods, nanorattles and toroids) and poly(N-isopropylacrylamide), PNIPAM, surface coating using the temperature-directed morphology transformation (TDMT) method. Globular PNIPAM (i.e. water insoluble) surface coatings have been shown to significantly increase cell uptake and enhanced enzyme activity. We incorporated a globular component of PNIPAM on the nanostructure surface, and examined the in vivo biodistribution of these nanostructures and accumulation in various tissues and organs in a mouse model. The in vivo biodistribution as a function of time was influenced by the shape and PNIPAM surface composition, in which organ capture and retention was the highest in the spleen. The rods (~150 nm in length and 15 nm in width) showed the highest capture and retention of greater than 35% to the initial injection amount compared to all other nanostructures. It was found that the rods specifically targeted the cells in the red pulp region of the spleen due to the shape and PNIPAM coating of the rod. This remarkable accumulation and selectively into the spleen represents new nanoparticle design parameters to develop new splenotropic effects for vaccines and other therapeutics.

Modulation of the immune response is an important step in the induction of protective humoral and cellular immunity against pathogens. In this study, we investigated the possibility of using a nanomaterial conjugated with the toll-like receptor (TLR) ligand CpG to modulate the immune response towards the preferred polarity. MgAl-layered double hydroxide (LDH) nanomaterial has a very similar chemical composition to Alum, an FDA approved adjuvant for human vaccination. We used a model antigen, ovalbumin (OVA) to demonstrate that MgAl-LDH had comparable adjuvant activity to Alum, but much weaker inflammation. Conjugation of TLR9 ligand CpG to LDH nanoparticles significantly enhanced the antibody response and promoted a switch from Th2 toward Th1 response, demonstrated by a change in the IgG2a:IgG1 ratio. Moreover, immunization of mice with CpG-OVA-conjugated LDH before challenge with OVA-expressing B16/F10 tumor cells retarded tumor growth. Together, these data indicate that LDH nanomaterial can be used as an immune adjuvant to promote Th1 or Th2 dominant immune responses suitable for vaccination purposes.

Melanoma, the most life-threatening skin cancer, requires more effective therapies. Methodology: A new folic acid (FA) receptor-targeted lipid-coated calcium carbonate/phosphate (LCCP) nanoparticle was synthesized, incorporating two often-used therapeutics, cell death siRNA and α-tocopheryl succinate. Results: The nanoparticles were spherical, with an average size of 40 nm. The nanoparticles exhibited a high gene/drug loading efficiency (60%), with folic acid-enhanced cellular uptake. The nanoparticles with both therapeutics enhanced inhibition of B16F0 melanoma cell growth, showing a moderate synergistic effect. The mechanism of the inhibition is associated with induction of cell apoptosis and cell cycle arrest at G1 phase. Conclusion: Our data indicate that lipid-coated calcium carbonate/phosphate nanoparticles are a potential platform for targeted therapy for melanoma.

The treatment of cartilage defects poses a clinical challenge owing to the lack of intrinsic regenerative capacity of cartilage. The use of tissue engineering techniques to bioengineer articular cartilage is promising and may hold the key to the successful regeneration of cartilage tissue. Natural and synthetic biomaterials have been used to recreate the microarchitecture of articular cartilage through multilayered biomimetic scaffolds. Acellular scaffolds preserve the microarchitecture of articular cartilage through a process of decellularization of biological tissue. Although promising, this technique often results in poor biomechanical strength of the graft. However, biomechanical strength could be improved if biomaterials could be incorporated back into the decellularized tissue to overcome this limitation.

The seasonal outbreaks of human rotavirus (RV) infection occur every winter. Most patients are diagnosed clinically by a rapid latex agglutination detection kit or polymerase chain reaction assays for RV from stool samples, but some problems have been reported on the specificity and sensitivity of such rapid detection assays. To ratify these issues, a sensitive, specific, simple, and rapid nucleic acid based diagnostic method is expected to be introduced and the reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect the RV in human stool samples by incubation at 60 °C for 1 h and amplification was confirmed by electrophoretic laddering, restriction enzyme digestion, and hydroxynapthol blue discoloration. The assay established in this study was found to detect only the RVs and no cross-reaction with other viruses, demonstrating its high specificity. By using serial samples dilution as template, the detection limit of LAMP was 10 times more than...

Movement of skeletal-muscle fibers is generated by the coordinated action of several cells taking part within the locomotion circuit (motoneurons, sensory-neurons, Schwann cells, astrocytes, microglia, and muscle-cells). Failures in any part of this circuit could impede or hinder coordinated muscle movement and cause a neuromuscular disease (NMD) or determine its severity. Studying fragments of the circuit cannot provide a comprehensive and complete view of the pathological process. We trace the historic developments of studies focused on in-vitro modeling of the spinal-locomotion circuit and how bioengineered innovative technologies show advantages for an accurate mimicking of physiological conditions of spinal-locomotion circuit. New developments on compartmentalized microfluidic culture systems (cµFCS), the use of human induced pluripotent stem cells (hiPSCs) and 3D cell-cultures are analyzed. We finally address limitations of current study models and three main challenges on neuromuscular studies: (i) mimic the whole spinal-locomotion circuit including all cell-types involved and the evaluation of independent and interdependent roles of each one; (ii) mimic the neurodegenerative response of mature neurons in-vitro as it occurs in-vivo; and (iii) develop, tune, implement, and combine cµFCS, hiPSC, and 3D-culture technologies to ultimately create patient-specific complete, translational, and reliable NMD in-vitro model. Overcoming these challenges would significantly facilitate understanding the events taking place in NMDs and accelerate the process of finding new therapies.

Epidemiological and genetic studies provide strong evidence supporting an association between myopia and glaucoma. The accurate detection of glaucoma in myopic eyes, especially those with high myopia, remains clinically challenging due to characteristic morphologic features of the myopic optic nerve in addition to limitations of current optic nerve imaging modalities. Distinguishing glaucoma from myopia is further complicated by overlapping perimetric findings. Therefore, longitudinal follow-up is essential to differentiate progressive structural and functional abnormalities indicative of glaucoma from defects that may result from myopia alone. Highly myopic eyes are at increased risk of complications from traditional incisional glaucoma surgery and may benefit from newer microinvasive glaucoma surgeries in select cases.

Regenerative medicine represents a rapidly evolving field that harnesses the body's biological mechanisms to repair, replace, or regenerate cells, tissues, and organs. This review focuses on its clinical applications in musculoskeletal disorders, with specific attention to four primary therapeutic modalities: viscosupplementation, prolotherapy, platelet-rich plasma (PRP), and mesenchymal stem cell (MSC) therapy. These therapies offer non-surgical options for conditions such as osteoarthritis (OA), tendinopathy, ligament injuries, and spinal pathologies. Viscosupplementation, through intra-articular injection of hyaluronic acid, improves joint lubrication and reduces pain in OA. Prolotherapy utilizes hypertonic dextrose to stimulate local inflammation and tissue regeneration. PRP therapy delivers concentrated platelets to injury sites, releasing growth factors that enhance healing. MSCs, obtained from bone marrow, adipose tissue, or umbilical sources, are multipotent cells capable of differentiating into musculoskeletal tissues. This article explores the techniques, indications, contraindications, clinical outcomes, and complications of these therapies, while emphasizing the role of interprofessional collaboration in optimizing treatment efficacy and patient safety. Despite promising outcomes, variability in preparation protocols and a lack of standardized guidelines necessitate further high-quality research to validate long-term benefits and establish clinical standards.