Review: Neuronal Differentiation Protocols of Mesenchymal Stem Cells
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Abstract
Mesenchymal stem cells (MSCs) are self-renewing cells found in almost all
postnatal organs and tissues in the perivascular region. These cells present
multiple characteristics that make them candidates to be applied in cell therapy for neurodegenerative diseases, such as their secretory action, migration
to the lesion area, and immunomodulatory potential. These cells have a high
capacity for mesodermal differentiation; however, numerous studies have
shown that MSCs can also differentiate into neurons. However, despite positive results in multiple trials in which undifferentiated MSCs transplanted
into animal models of neurodegenerative diseases, some studies suggest that
the therapeutic effects obtained are enhanced by the use of MSCs differentiated towards the neuronal lineage before transplant. In this sense, there are
several methods to induce in vitro reprogramming of MSCs towards the neuronal lineage, including chemical substances, growth factors, cocultures with
neural lineage cells, transfection of genes, miRNAs, etc., and small molecules
stand out. Therefore, this article compares multiple experimental tests in which
these inducers promote neuronal differentiation of MSCs and identify those
methods that originate an optimal neuronal differentiation. The analysis includes the percentage of differentiation, maturation, expression of neuronal
markers, functionality, and cell survival considering the intrinsic characteristics of the MSCs used as the tissue of origin and the species from which they
were isolated.
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Miguel A. Jiménez Acosta,
Lory Jhenifer Rochín Hernández,
Mayte Lizeth Padilla Cristerna,
Marco A. Meraz Ríos,
José Tapia Ramírez,
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Effects of Heavy Metal Ions (Fe3+, Cu2+, Zn2+ and Cr3+) on the Productivity of Biogas and Biomethane Production
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Abstract
It was shown that the concentration of heavy metal ions Fe3+, Cu2+, Cr3+ and Zn2+, which are contained in the fermentation medium, affects the process of biogas production. The aim of this paper was to establish the concentration of ions, which makes it possible to increase the yield of biogas and the methane content in it. The total yield of biogas per unit of dry organic matter, methane and the kinetics of changes in the yield of biogas during fermentation of cattle manure in the presence of different concentrations of metal ions in the anae robic environment. It was established that the content of Fe3+, Cu2+ is up to 80 mg/dm3, Cr3+ and Zn2+ is up to 50 mg/dm3 in the medium lead to increased methane production by anaerobic association of microorganisms due to the effect on the activity of enzyme systems in comparison with the sample without the addition of metal ions. It was found that the rational concentra tions to increase the methane yield in the biogas obtained by fermentation of cattle manure are: Fe3+—20 - 40 mg/dm3, Cu2+—40 - 60 mg/dm3 , Cr3+—10 mg/dm3. The increase in the concentration of metal ions above rational values leads to a decrease in the methane content in biogas. It was shown that zinc ions have a positive effect on methane production, but reduce the total biogas yield and, accordingly, the degree of conversion of organic raw materials. Therefore, the rational concentration of Zn2+ in the fermentation medium is 10 mg/dm3.
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N. B. Golub,
A. V. Shynkarchuk,
O. A. Kozlovets,
M. V. Kozlovets,
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Effect of Indigenous Phytase-Producing Yeast Cultures on Growth Performance, Digestion and Health of Rabbits (Oryctolagus cuniculus)
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Abstract
This study investigated the effects of dietary supplementation of indigenous phytase producing live yeast cultures on growth performance, phytate degradation and blood biochemical parameters of weanling rabbits. Fifty-six rabbits (28 males and 28 females) were allotted randomly into four groups containing a control and three groups administrated each with the yeasts Saccharomyces cerevisiae ADR1B1, Hanseniaspora jakobsenii ADR3E1, Hanseniaspora guilliermondii RD31 during 8 weeks. The results showed that the administration of the three yeast strains accelerated the weight gain and improved rabbits growth performance in comparison to the control group, but did not affect the serum biochemical and hematological parameters. The highest weight gain (1715.95 ± 236 g), and phytate degradation rate (73.82%± 0.92%) and the lowest biochemical parameters (triglycerides, cholesterol, HDL and LDL) were observed on rabbits fed with the yeast Hanseniaspora jakobsenii ADR3E1. This indigenous yeast species, which is used for the first time in animal feeding provided the best beneficial effects in rabbit breeding. Therefore, based on this finding, Hanseniaspora jakobsenii ADR3E1 can be recommended to supplement rabbit diets for growth performance and profitability enhancement.
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N’sa Koky Marc Celaire,
Ake Moussan Désirée Francine,
Dje Koffi Marcellin,
Djeni N’Dede Theodore,
Kimse Moussa,
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2022 |
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Evaluation of Red Blood Cell Contribution to Platelet Activation in the Bulk Applying Red Blood Cell—Platelet Thrombus as a Point Source Model
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Abstract
In this study, a point source mathematical model is proposed to describe the diffusion of adenosine di-phosphate (ADP) from either damaged red blood cell (RBC) or activated platelet. The convective diffusion equation is reduced
to describe the suggested problem. The final differential equation is solved using Laplace transforms and ADP concentration profiles around the source are obtained. Thrombi of 5 to 20 µm3 containing platelets and a range of red
blood cells (RBCs) (0%, 25%, 50%, 75%, 100%) concentrations are used to apply the model. Reported ADP concentrations in the literature are used and its dynamic release from the point source is calculated. Results suggest that RBC chemical contribution to platelet aggregation in the bulk is much less than that of platelet (almost) negligible. However, the physical effect of RBCs is dominant in the bulk through augmentation of released ADP and platelets
diffusivities. Moreover, the chemical contribution reported in previous studies is suggested to be as a result of interaction of RBC with the surface under the influence of shear stresses in the boundary region.
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Taha M. Alkhamis,
Bahieh M. Almaatah,
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2022 |
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Assessments of Antibacterial and Antioxidant Properties in the Methanolic and Aqueous Leaf Extracts of Pistacia lentiscus against Different Antibiotic Resistance Pathogenic Bacteria
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Abstract
The current study was carried out to determine the bioactivity of P. lentiscus leaf extracts as potential antibacterial and antioxidant properties. The plant extracts were examined for antibacterial activity against antibiotic-resistant
Staphylococcus aureus, Staphylococcus haemolyticus, Pseudomonas aeruginosa, and Proteus mirabilis using the agar well method (according to the guidelines of Clinical and Laboratory Standard Institute). The antioxidant
potential of 3 plant leaf extracts was determined by their ability to convert Fe3+ to Fe2+ and scavenge the DPPH free radical. At all concentrations studied, the methanolic leaf extract had higher total phenolic and flavonoid content, as well as stronger antioxidant and antibacterial inhibitory activity compared to aqueous extract. Our findings with P. aeruginosa were especially interesting, because this bacterium was inhibited by methanol extract than that
of the reference antibiotics. The results also demonstrated a link between DPPH radical scavenging ability, reducing power, and total phenolic and flavonoid content of plant extracts (r > 0.97, R2 > 0.95, P = 0.01). As a result, the
methanolic leaf extract of the chosen plant might be employed as an effective antioxidant and antibacterial agent for the treatment of a variety of morbidities.
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Aml O. Alhadad,
Galal S. Salem,
Suliman M. Hussein,
Sarah M. Elshareef,
Maraia F. Elmhdwi,
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2022 |
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Identification Transgenic DNA of Transformed Arabidopsis Using PCR and Southern Blot
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Abstract
For identification of transgenic DNA of Arabidopsis plant material, the followed methods are transformation and growing of seeds after bleaching them for sterilization should be performed then using polymerase chain reaction
(PCR) for amplification the gene before making gel electrophoresis to be ready for southern blot analysis, this method is possible because the length of the given primers, there are slightly modifications in the methods as ligating PCR
technique developed for amplification of the unknown DNA after extracting it to determine its transgenicity. The methods were specific and reproducible for many different plants from transgenic and non-transgenic cell lines. Furthermore, the number of products of PCR result can be considered as a good estimation of transgenic DNA, during comparison to southern blot analysis, the results obtained by the PCR give information about DNA of Arabidopsis,
applications of the approach to Arabidopsis plants after transformation with an Agrobacterium-mediated transformation will be described.
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Al-Baraa Akram,
Glen McCann,
Triaziana Sgamma,
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2022 |
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Engineering Microbial Biofilms for Improved Productivity of Biochemicals Important in Restoration of Degraded Ecosystems
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Abstract
Biofilms are being engineered in-vitro to produce numerous commodities like biofertilizers, pharmaceuticals, biofuels and electricity, the efficacies of which rely on the biochemicals secreted by the biofilms i.e. extracellular polymeric
substances (EPS). It has been shown that once EPS-biochemicals of developed biofilms are applied to an ecosystem, they can restore degraded complex ecosystem networks for improved ecosystem functioning and sustainability.
Identification of the EPS biochemicals and understanding their contributions to the network interactions in particular, are at initial stage. In the present study, using Aspergillus niger, Nostoc sp., and gram (−) Stenotrophomonas
maltophilia & gram (+) Bacillus subtilis as test fungal (F), cyanobacterial (C), and bacterial (B) counterparts, respectively we analyzed morphology and biochemical parameters of fungal-bacterial (FBBs), fungal-cyanobacterial (FCBs), cyanobacterial-bacterial (CBBs), and fungal-cyanobacterial-bacterial biofilms (FCBBs). Results revealed that the FCBBs produced the highest concentrations of lipids, proteins, and polysaccharides whereas FBBs generated the
highest diversity of biochemicals. Bacterial type (i.e. gram + or −) and microbial composition in the biofilm affected the biochemical production. Ecologically and industrially important diverse biochemicals which are used individually as medicines, bioremediating agents and industrial chemicals in human society with certain adverse and beneficial effects were detected in the biofilm-EPS. However, in the nature, simultaneous action of those diverse biochemicals applied as biofertilizers has already shown a huge potential to restore the entire agroecosystems degraded due to farmers’ detrimental practices. This striking difference in utilization of the biochemicals and their enhanced effect when they act simultaneously needs further investigations for their better applications.
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Mahesh Premarathna,
Thilini Rathnathilaka,
Gamini Seneviratne,
Sumedha Madawala,
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2022 |
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Simulating Microbial Functional Diversity Dynamics in Agricultural Soils: An Individual Based Modeling Approach
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Abstract
New agricultural soil model approaches based on the microbiome dynamics must be considered as they can contribute to understand microbiological soil processes directly linked to substrate metabolism and the influence of these
processes on plant growth. The present work presents an approach to the modelling of the interactions of the soil microbial functional diversity with the plant in terms of functions associated to specific processes of organic
Carbon and Nitrogen metabolism. The substrates transformations arising in the organic matter that enters as a part of an agricultural scheme are the base for define this metabolism. As result, it has been possible to simulate a rhizospheric soil based on the concept of complex system dynamics and Individual Based Modeling known too as Agent-Based Modeling in an agricultural management context. The explicit definition of the microbiome functional
diversity and the processing of the structural elements Carbon and Nitrogen, allowed representing the functional dynamics of this complex system composed by microorganisms, Carbon, Nitrogen and the plant. The variables that
reflect the biology and the adaptation to the rhizospheric environment characterized the microorganisms and the assemblage community patterns in time. The main simulations output are system glucose and nitrate levels and
an approach to plant growth, all resultant from the metabolic process of the considered Carbon and Nitrogen consortia. The results indicate that the microorganism’s diversity assemblages and its functional expression have a fundamental role in terms of plant growth.
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Diego Rubio Fernandez,
Emiliano Barreto Hernández,
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2022 |
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Spectrophotometric Assay for the Quantification of Plasma Ethanol Levels in Mice through Chromium-Ethanol Oxidation-Reduction Reaction
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Abstract
The quantification of blood/plasma ethanol concentration (BEC/PEC) is of great importance in experiments involving basic research, clinical studies, and bioethanol production. Traditional methods commonly used to measure
BEC can be expensive and require high-cost equipment and qualified labor. The aim of this study was to develop a low-cost method that can be performed with simple infrastructure commonly available in research laboratories. For this, we developed a protocol to quantify PEC in mice, using the method of reduction of potassium dichromate by ethanol. However, this oxidation-reduction (redox) reaction is not specific to ethanol. Thus, the PEC was measured following a sequence of chemical reactions to eliminate the reductive interfering substances presented in the samples. Firstly, we evaluated the sensitivity of the dichromate reactive to ethanol and to different reducing substances found in the plasma, in order to determine which the main interfering substances are. Next, once the main interfering substances were determined in the dichromate reduction, plasma was assayed for PEC. First, mice received intraperitoneally (i.p.) saline (basal reading, 0% ethanol) or ethanol injections (0.5, 1, 2, 3, and 4 g/kg) and had their plasma collected. After plasma deproteinization and plasma glucose oxidation, it was mixed with the dichromate/acetic acid reactive, and then the products of the redox reaction were determined by the spectrophotometric method. Then, we determined the PEC with the same plasma samples using a commercial ethanol assay kit as a positive control. We found an excellent correlation between the administered ethanol doses and PECs in both the methods analyzed. The values of PEC found in the dichromate reaction method were similar to those obtained in the literature with the same ethanol doses, and to the commercial enzyme activity assay. Therefore, despite the need for a background reading, this method can be successfully applied to determine PEC using low-cost chemical reagents.
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Marcos Brandão Contó,
Rosana Camarini,
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2022 |
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Anti-Angiogenic and Anti-Metastatic Effects of Biogenic Silver Nanoparticles Synthesized Using Azadirachta indica
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Abstract
Nanotechnology symbolizes a broad discipline with enormous potential in cancer treatment bridging one of the bottlenecks of traditional approaches in cancer therapy which is an inability to deliver adequate quantities of anti-cancer drugs to the tumor area. Studies on nanoparticles indicate their importance in cancer angiogenesis and metastasis. Aim: The present study assessed anti-angiogenesis and anti-metastatic effects of biogenic silver
nanoparticles (AgNPs) synthesized from neem plant (Azadirachta indica). Methods: Chicken chorioallantoic membrane (CAM) and two-dimensional (2D) wound healing assays were used to study anti-angiogenic and anti-metastatic effects of the AgNPs respectively. Twenty-four fertilized eggs were divided into four groups: two biogenic AgNPs treatments at 100 µg/ml and 200 µg/ml; negative control (1% DMSO) and positive control (cyclophosphamide). On day 8 of incubation, filter discs impregnated with different concentration levels of the treatments were placed on the CAM. On day 12 of incubation, the CAMs were imaged using a stereomicroscope, scaled using ImageJ, and different morphometric and spatial parameters computed using AngioTool software. Vessel area, vessel percent area, total number of junctions, total vessel length, average vessel length, mean lacunarity, and junction density were measured. The crown-rump length (CRL) and fetal weight were also recorded on day 16 of incubation. In order to determine relative gene expression profiles of iNOS and VEGF, total RNA was extracted from the CAM, and qRT-PCR was performed with β-Actin as a reference gene. For the 2D wound healing assay, DU145 human prostate cells were grown in Dulbecco’s Modified Eagle’s Medium supplemented with 10% Fetal Bovine Serum. Results:Biogenic AgNPs demonstrated anti-angiogenic effects in a dose-dependent manner in the parameters generated from the CAM images. Also, qRT-PCR revealed down-regulation of iNOS and VEGF genes. The 2-dimensional wound healing assay showed inhibition of migration and motility of the DU145 cells for the 72-hours of assessment. Conclusion: The present study postulates that the biogenic AgNPs can prevent angiogenesis by inactivation of VEGF-NO and VEGF/VEGF-R pathways while inhibiting cell migration and metastasis.
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Ahmed A. Abdille,
Judith Sokei,
Shedrack Reuben Kitimu,
Peter Kirira,
Dominic Ochwang,
Peter Mwitari,
Naomi Maina,
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2022 |
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Expression of miR-155 and miR-146a in Skin Tissue, Peripheral Blood Mononuclear Cells and Serum of Patients with Psoriasis and Its Clinical Significance
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Abstract
Objective: To explore the expression and clinical significance of miR-155 and miR-146a in psoriasis patients from different sources. Methods: The expression levels of miR-155 and miR-146a in psoriasis vulgaris tissues, peripheral
blood mononuclear cells, and serum were detected by the QRT-PCR method in 30 patients with psoriasis vulgaris and 30 healthy subjects as the normal control group, and the correlation between the expression levels of miR-155
and miR-146a in psoriasis samples from different sources was analyzed. And the expression levels of miR-155 and miR-146a in peripheral blood mono nuclear cells, and serum after treatment. Results: The expression of miR-155
and miR-146a in skin tissue, peripheral blood mononuclear cells and serum of patients with psoriasis was significantly higher than that of normal controls (P < 0.01). miR-155 and miR-146a were highly expressed in skin tissue, peripheral blood mononuclear cells and serum of the psoriasis experimental group (P < 0.01). After treatment, the expression of miR-155 and miR-146a in peripheral blood mononuclear cells and serum decreased (P < 0.05). Conclusions: The significantly increased levels of miR-155 and miR-146a in psoriasis samples from different sources may mediate the immune mechanism involved in the activity process of psoriasis.
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Dongcheng Gu,
Jurui Feng,
Maojie Cheng,
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2022 |
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