Microinstrumentation for Brain Organoids.

Brain organoids are three-dimensional aggregates of self-organized differentiated stem cells that mimic the structure and function of human brain regions. Organoids bridge the gaps between conventional drug screening models such as planar mammalian cell culture, animal studies, and clinical trials. They can revolutionize the fields of developmental biology, neuroscience, toxicology, and computer engineering. Conventional microinstrumentation for conventional cellular engineering, such as planar microfluidic chips; microelectrode arrays (MEAs); and optical, magnetic, and acoustic techniques, has limitations when applied to three-dimensional (3D) organoids, primarily due to their limits with inherently two-dimensional geometry and interfacing. Hence, there is an urgent need to develop new instrumentation compatible with live cell culture techniques and with scalable 3D formats relevant to organoids. This review discusses conventional planar approaches and emerging 3D microinstrumentation necessary for advanced organoid-machine interfaces. Specifically, this article surveys recently developed microinstrumentation, including 3D printed and curved microfluidics, 3D and fast-scan optical techniques, buckling and self-folding MEAs, 3D interfaces for electrochemical measurements, and 3D spatially controllable magnetic and acoustic technologies relevant to two-way information transfer with brain organoids. This article highlights key challenges that must be addressed for robust organoid culture and reliable 3D spatiotemporal information transfer.

A novel approach to increase glial cell populations in brain microphysiological systems.

Brain microphysiological systems (bMPS), which recapitulate human brain cellular architecture and functionality more closely than traditional monolayer cultures, have become a practical, non-invasive, and increasingly relevant platform for the study of neurological function in health and disease. These models include 3D spheroids and organoids as well as organ-on-chip models. Currently, however, existing 3D brain models vary in reflecting the relative populations of the different cell types present in the human brain. Most of the models consist mainly of neurons, while glial cells represent a smaller portion of the cell populations. Here, by means of a chemically defined glial-enriched medium (GEM), we present an improved method to expand the population of astrocytes and oligodendrocytes without compromising neuronal differentiation in bMPS. An important finding is that astrocytes not only increased in number but also changed in morphology when cultured in GEM, more closely recapitulating primary culture astrocytes. We demonstrate oligodendrocyte and astrocyte enrichment in GEM bMPS using a variety of complementary methods. We found that GEM bMPS are electro-chemically active and showed different patterns of Ca +2 staining and flux. Synaptic vesicles and terminals observed by electron microscopy were also present. No significant changes in neuronal differentiation were observed by gene expression, however, GEM enhanced neurite outgrowth and cell migration, and differentially modulated neuronal maturation in two different iPSC lines. Our results have the potential to significantly improve in vivo-like functionality of bMPS for the study of neurological diseases and drug discovery, contributing to the unmet need for safe human models.

Farming Sparus aurata (Teleostei: Sparidae) in marsh ponds: trophic characterization and trace metal accumulation.

Considering the overexploitation of fishing on most of the world coasts, the ingestion of fish and shellfish will depend mostly on aquaculture production. Since intensive mariculture usually involves environmental impact, developing sustainable cultures is a priority. In this sense, salt marshes can provide ecosystem services and incorporate both conservation and extensive aquaculture activities. In the present study we compared gilthead seabream Sparus aurata Linnaeus, 1758 cultured in extensive and semi-intensive marsh ponds with wild conspecifics from surrounding coastal areas, using trophic characterization (diet analysis and stable isotopes) and trace metal accumulation. Stomach content analysis revealed different feeding habits among gilthead seabream from different origin. Although wild specimens had the most diverse diet, results of stable isotopes showed that extensive diet had the wider isotopic niche and revealed the highest similarities between wild and extensively cultured gilthead seabream. A similar trace element signature was also measured in wild and extensive culture, whereas the semi-intensive culture showed different concentrations for several elements. Cr, Fe and Mn showed the highest concentrations in semi-intensive cultured fish, while As and Zn showed the lowest values in this group. In any case, average values measured in both extensive and semi-intensive culture were, in general terms, below the hazardous limits provided by the standards recommended for trace metals by national and international regulations. Therefore, marsh ponds provide a suitable environment where the cultured fish, especially extensive, should be promoted.

Single-cell transcriptomic reveals molecular diversity and developmental heterogeneity of human stem cell-derived oligodendrocyte lineage cells.

Injury and loss of oligodendrocytes can cause demyelinating diseases such as multiple sclerosis. To improve our understanding of human oligodendrocyte development, which could facilitate development of remyelination-based treatment strategies, here we describe time-course single-cell-transcriptomic analysis of developing human stem cell-derived oligodendrocyte-lineage-cells (hOLLCs). The study includes hOLLCs derived from both genome engineered embryonic stem cell (ESC) reporter cells containing an Identification-and-Purification tag driven by the endogenous PDGFRα promoter and from unmodified induced pluripotent (iPS) cells. Our analysis uncovers substantial transcriptional heterogeneity of PDGFRα-lineage hOLLCs. We discover sub-populations of human oligodendrocyte progenitor cells (hOPCs) including a potential cytokine-responsive hOPC subset, and identify candidate regulatory genes/networks that define the identity of these sub-populations. Pseudotime trajectory analysis defines developmental pathways of oligodendrocytes vs astrocytes from PDGFRα-expressing hOPCs and predicts differentially expressed genes between the two lineages. In addition, pathway enrichment analysis followed by pharmacological intervention of these pathways confirm that mTOR and cholesterol biosynthesis signaling pathways are involved in maturation of oligodendrocytes from hOPCs.

Cardiovascular risk factors associated with coronary ectasia and acute myocardial infarction.

INTRODUCTION: Coronary ectasia has a low prevalence in the general population. Its risk factors may differ from those of coronary artery disease. OBJECTIVE: To identify the prevalence of coronary ectasia in patients with acute myocardial infarction (AMI) and cardiovascular risk factors (CVRFs). METHODS: Retrospective, cross-sectional study. Out of 3,254 cardiac catheterizations for AMI during one year, 2,975 had no coronary ectasia. We included 558 patients with coronary ectasia on coronary angiography and, as controls, subjects with similar characteristics except for coronary ectasia, and CVRFs were recorded. Descriptive statistics, bivariate and multivariate analysis were used; odds ratio (OR) was calculated. RESULTS: 279 patients with and without coronary ectasia were studied. The prevalence of coronary ectasia was 8.5 %. The platelet/lymphocyte ratio (PLR) was higher in patients with ectasia than in those without ectasia (p = 0.003). In the bivariate analysis, associated CVRFs were overweight, obesity and diabetes, and in the multivariate analysis, hypercholesterolemia (OR: 3.90; p = 0.0001) and exposure to herbicides (OR: 6.82; p = 0.020). CONCLUSIONS: A high prevalence of coronary ectasia was found, with the main risk factors being a history of herbicide use and hypercholesterolemia. PLR was found to be elevated in these patients. Early detection is important due to its association with acute coronary events.

INTRODUCCIÓN: La ectasia coronaria tiene baja prevalencia en población general, los factores de riesgo pueden diferir de la enfermedad arterial coronaria. OBJETIVO: Identificar la prevalencia de ectasia coronaria en pacientes con infarto agudo de miocardio (IAM) y factores de riesgo cardiovascular (FRCV). MÉTODOS: Estudio retrospectivo, transversal. De 3,254 cateterismos cardiacos por IAM durante un año, 2,975 no presentaron ectasia coronaria. Se incluyeron 558 pacientes clasificados como portadores de ectasia coronaria en coronariografía y controles aquellos con características similares exceptuando la ectasia coronaria y se registraron los FRCV. Empleamos estadística descriptiva, análisis bivariante, multivariante y calculamos el odds ratio (OR). RESULTADOS: Se estudiaron 279 pacientes con y sin ectasia coronaria. La prevalencia de ectasia coronaria fue del 8.5%. El índice plaqueta/linfocito (IPL) se encontró más elevado en pacientes con ectasia que en aquellos sin ectasia (p = 0.003). En el análisis bivariante los FRCV asociados fueron sobrepeso, obesidad y diabetes, y en el multivariante la hipercolesterolemia (OR: 3.90; p = 0.0001) y exposición a herbicidas (OR: 6.82; p = 0.020). CONCLUSIONES: Encontramos alta prevalencia de ectasia coronaria, los principales factores de riesgo fueron el antecedente de uso de herbicidas e hipercolesterolemia. Identificamos el IPL elevado en estos pacientes. Es importante la detección oportuna debido a su asociación con eventos coronarios agudos.

Design of a Helical-Stabilized, Cyclic, and Nontoxic Analogue of the Peptide Cm-p5 with Improved Antifungal Activity.

Following the information obtained by a rational design study, a cyclic and helical-stabilized analogue of the peptide Cm-p5 was synthetized. The cyclic monomer showed an increased activity in vitro against Candida albicans and Candida parapsilosis, compared to Cm-p5. Initially, 14 mutants of Cm-p5 were synthesized following a rational design to improve the antifungal activity and pharmacological properties. Antimicrobial testing showed that the activity was lost in each of these 14 analogues, suggesting, as a main conclusion, that a Glu-His salt bridge could stabilize Cm-p5 helical conformation during the interaction with the plasma membrane. A derivative, obtained by substitution of Glu and His for Cys, was synthesized and oxidized with the generation of a cyclic monomer with improved antifungal activity. In addition, two dimers were generated during the oxidation procedure, a parallel and antiparallel one. The dimers showed a helical secondary structure in water, whereas the cyclic monomer only showed this conformation in SDS. Molecular dynamic simulations confirmed the helical stabilizations for all of them, therefore indicating the possible essential role of the Glu-His salt bridge. In addition, the antiparallel dimer showed a moderate activity against Pseudomonas aeruginosa and a significant activity against Listeria monocytogenes. Neither the cyclic monomer nor the dimers were toxic against macrophages or THP-1 human cells. Due to its increased capacity for fungal control compared to fluconazole, its low cytotoxicity, together with a stabilized α-helix and disulfide bridges, that may advance its metabolic stability, and in vivo activity, the new cyclic Cm-p5 monomer represents a potential systemic antifungal therapeutic candidate.

Presence of a predator image in potential breeding sites and oviposition responses of a dengue vector.

In dengue vector control, attempts to minimize or replace the use of pesticides have mostly involved use of predators, but success has been severely impeded by difficulties associated with financial and environmental costs, predator mass production, and persistence in target habitats. Visual deterrents have been used successfully to control animal pests, in some cases in an effort to replace pesticide use. Despite evidence that visual signals are crucial in site choice for egg deposition by dengue vectors, and that female mosquitoes respond to artificial predation, the role of predator intimidation as it affects the oviposition behavior of dengue vectors remains largely unexplored. Here, we examined the oviposition responses of Aedes aegypti exposed to various mosquito predator pictures. Gravid females were presented with equal opportunities to oviposit in two cups with predator images [Toxorhynchites splendens-TXI, Goldfish (Carassius auratus)-small (SFI) and large (LFI) and Tx. splendens+Goldfish-TXFI] and two others without pictures. Differences in egg deposition were examined between sites with and without these images. When given a chance to oviposit in cups with and without TXI, Ae. aegypti females were similarly attracted to both sites. When provided an opportunity to oviposit in cups displaying pictures of fish (SFI or LFI) and blank cups, egg deposition rates were much lower in the fish picture sites. Females showed a preference for blank cups over TXFI for egg deposition. They also equally avoided cups with pictures of fish, regardless of the size of the picture. Our results indicate that the presence of images of goldfish and their association with Tx. larvae significantly reduced egg deposition by Ae. aegypti, and this was not the case with the predatory larvae alone. The observations that the images of natural predators can repel gravid females of a dengue vector provide novel possibilities to develop effective and inexpensive alternative tools to harmful insecticides.

Biomechanics of human parietal pleura in uniaxial extension.

Tension pneumothorax, a major preventable cause of battlefield death, often arises from chest trauma and is treated by needle decompression to release trapped air from the pleural cavity. Surgical simulation mannequins are often employed to train medical personnel to perform this procedure properly. Accurate reproduction of the mechanical behavior of the parietal pleura, especially in response to needle penetration, is essential to maximize the fidelity of these surgical simulators. To date, however, the design of pleura-simulating material has been largely empirical and based on subjective practitioner feel rather than on the tissue properties, which have remained unknown. In this study, we performed uniaxial extension tests on samples of cadaveric human parietal pleura. We found that the pleura was highly nonlinear and mildly anisotropic, being roughly twice as stiff in the direction parallel to the ribs vs. perpendicular to the ribs (large-strain modulus = 20.44 vs. 11.49MPa). We also did not find significant correlations for most pleural properties with age or BMI, but it must be recognized that the age range (59 ± 9.5 yrs) and BMI range (31 ± 5.3) of the donors in our study was not what one might expect from combatants, and there could be differences for younger, lighter individuals. We found a significantly higher low-strain modulus in the diabetic donors (0.213 vs. 0.100MPa), consistent with the general tendency of tissue to stiffen in diabetes. The nonlinearity and tensile strength should be considered in material design and selection for future surgical simulators.

Microondas. Experimentación Pura y clínica / Microwave. Pura experimental and clinical

Las microondas constituyen una subfamilia de la gran gama de ondas electromagnéticas que existen en el universo. Utilizamos dichas microondas en una frecuenciade 2,450 MHz como sustancia imponderable, obtenida de un magnetrón de un horno de microondas convencional, para la preparación de un medicamentohomeopático en las escalas cincuentamilesimal y centesimal por medio del método clásico. Llevamos a cabo una patogenesia y luego su aplicación clínica paralograr una aproximación al conocimiento de los síntomas y signos involucrados en esta investigación. Participaron 20 sujetos, 16 en experimentación pura y 4 enexperimentación clínica, bajo el esquema propuesto por Samuel Hahnemann en el Organon de la medicina en su sexta edición, fungiendo como grupo de controlel 10% del total de los participantes con placebo. Como resultado se obtuvo una patogenesia en la cual los rubros de mente y extremidades se expresaron comolos más activos con 13% del total de los síntomas cada uno; fueron seguidos por el rubro estómago, con el 11% del total, posteriormente el rubro cabeza, con 8%, ydescendentemente el resto de rubros con menos del 7% del total de los síntomas de la investigación cada uno. (AU)

Microwaves are a subfamily of a long range of electromagnetic waves that exist in the universe. We used these microwaves on a 2,450 MHz frequency as an imponderable substance, obtained from a magnetron through a conventional microwave oven to prepare an Homeopathic medicine in the cincuentamilesimal and centesimal classic scale. We wanted to do this pure experimentation and then the clinical application to know on a better way the expressed signs and symptoms. There were 20 subjects, 16 as pure experimentation subjects and 4 as clinical research subjects using the classical method of pure experimentation developed by Samuel Hahnemann in the Organon of medicine, sixth edition. Our control placebo group was formed by the 10% of total research subjects. As a result we obtained a pathogenesia in which the more active items were mind and extremities with 13% of total symptoms each, then head item with 8% and the rest of the items by descending order with less of 7% of the total symptoms each. (AU)

Microondas: experimentación pura y clínica / Microwave: pura experimental and clinical

Las microondas constituyen una subfamilia de la gran gama de ondas electromagnéticas que existen en el universo. Utilizamos dichas microondas en una frecuenciade 2,450 MHz como sustancia imponderable, obtenida de un magnetrón de un horno de microondas convencional, para la preparación de un medicamentohomeopático en las escalas cincuentamilesimal y centesimal por medio del método clásico. Llevamos a cabo una patogenesia y luego su aplicación clínica paralograr una aproximación al conocimiento de los síntomas y signos involucrados en esta investigación. Participaron 20 sujetos, 16 en experimentación pura y 4 enexperimentación clínica, bajo el esquema propuesto por Samuel Hahnemann en el Organon de la medicina en su sexta edición, fungiendo como grupo de controlel 10% del total de los participantes con placebo. Como resultado se obtuvo una patogenesia en la cual los rubros de mente y extremidades se expresaron comolos más activos con 13% del total de los síntomas cada uno; fueron seguidos por el rubro estómago, con el 11% del total, posteriormente el rubro cabeza, con 8%, ydescendentemente el resto de rubros con menos del 7% del total de los síntomas de la investigación cada uno.

Microwaves are a subfamily of a long range of electromagnetic waves that exist in the universe. We used these microwaves on a 2,450 MHz frequency as an imponderable substance, obtained from a magnetron through a conventional microwave oven to prepare an Homeopathic medicine in the cincuentamilesimal and centesimal classic scale. We wanted to do this pure experimentation and then the clinical application to know on a better way the expressed signs and symptoms. There were 20 subjects, 16 as pure experimentation subjects and 4 as clinical research subjects using the classical method of pure experimentation developed by Samuel Hahnemann in the Organon of medicine, sixth edition. Our control placebo group was formed by the 10% of total research subjects. As a result we obtained a pathogenesia in which the more active items were mind and extremities with 13% of total symptoms each, then head item with 8% and the rest of the items by descending order with less of 7% of the total symptoms each.

Detection and characterization of enteric viruses in flood water from the 2011 thai flood.

Severe flooding, which is associated with numerous outbreaks of a wide range of infectious diseases, particularly those caused by enteric viruses, occurred in all areas of Thailand in 2011. To determine the prevalence of five human enteric viruses, namely enterovirus, rotavirus (RV), norovirus (NV), hepatitis A virus (HAV), and hepatitis E virus, in the flood water, 100 water samples were collected from flood-damaged areas in central Thailand. Viral RNA was extracted from concentrated samples and analyzed by RT-PCR and sequencing. NV was the most commonly detected pathogen in the tested samples (14%). RV and HAV were detected in 9% and 7% of samples, respectively. This study is the first to detect enteric viral genes in flood water in Thailand. Furthermore, it is the first to detect an NV gene in any type of environmental water in Thailand. These results provide useful information for estimating the risk of flood waterborne viral infection.