Comparative metabarcoding and biodiversity of gut-associated fungal assemblages of Dendroctonus species (Curculionidae: Scolytinae).

The genus Dendroctonus is a Holarctic taxon composed of 21 nominal species; some of these species are well known in the world as disturbance agents of forest ecosystems. Under the bark of the host tree, these insects are involved in complex and dynamic associations with phoretic ectosymbiotic and endosymbiotic communities. Unlike filamentous fungi and bacteria, the ecological role of yeasts in the bark beetle holobiont is poorly understood, though yeasts were the first group to be recorded as microbial symbionts of these beetles. Our aim was characterize and compare the gut fungal assemblages associated to 14 species of Dendroctonus using the internal transcribed spacer 2 (ITS2) region. A total of 615,542 sequences were recovered yielding 248 fungal amplicon sequence variants (ASVs). The fungal diversity was represented by 4 phyla, 16 classes, 34 orders, 54 families, and 71 genera with different relative abundances among Dendroctonus species. The α-diversity consisted of 32 genera of yeasts and 39 genera of filamentous fungi. An analysis of ß-diversity indicated differences in the composition of the gut fungal assemblages among bark beetle species, with differences in species and phylogenetic diversity. A common core mycobiome was recognized at the genus level, integrated mainly by Candida present in all bark beetles, Nakazawaea, Cladosporium, Ogataea, and Yamadazyma. The bipartite networks confirmed that these fungal genera showed a strong association between beetle species and dominant fungi, which are key to maintaining the structure and stability of the fungal community. The functional variation in the trophic structure was identified among libraries and species, with pathotroph-saprotroph-symbiotroph represented at the highest frequency, followed by saprotroph-symbiotroph, and saprotroph only. The overall network suggested that yeast and fungal ASVs in the gut of these beetles showed positive and negative associations among them. This study outlines a mycobiome associated with Dendroctonus nutrition and provides a starting point for future in vitro and omics approaches addressing potential ecological functions and interactions among fungal assemblages and beetle hosts.

Tuning plasmid DNA amounts for cost-effective transfections of mammalian cells: when less is more.

Transient gene expression (TGE) in mammalian cells is a well-known approach to the fast expression of recombinant proteins. The human cell line HEK (human embryonic kidney) 293F is widely used in this field, due to its adaptability to grow in suspension to high cell densities in serum-free media, amenability to transfection, and production of recombinant proteins in satisfactory quantities for functional and structural analysis. Amounts of plasmid DNA (pDNA) required in transfections for TGE remain high (usually 1 µg pDNA/mL, or even higher), representing a noticeable proportion of the overall cost. Thus, there is an economic need to reduce amounts of coding pDNA in TGE processes. In this work, amounts of both pDNA and transfecting agent used for TGE in HEK 293F cells have been explored in order to reduce them without compromising (or even improving) the productivity of the process in terms of protein yield. In our hands, minimal polyethyleneimine (PEI) cytotoxicity and optimum protein yields were obtained when transfecting at 0.5 µg pDNA/mL (equal to 0.5 µg pDNA/million cells) and a DNA-to-PEI ratio of 1:3, a trend confirmed for several unrelated recombinant proteins. Thus, carefully tuning pDNA and transfecting agent amounts not only reduces the economic costs but also results in higher recombinant protein yields. These results surely have a direct application and interest for the biopharmaceutical industry, always concerned in increasing productivity while decreasing economic costs. KEY POINTS: • Mammalian cells are widely used to produce recombinant proteins in short times. • Tuning DNA and transfecting agent are of great interest to optimize economic costs. • Reducing DNA and transfecting agent amounts result in higher protein yields.

Conocimiento, creencias y aceptabilidad de la vacuna contra el Virus del Papiloma Humano en padres de Chihuahua, México / Knowledge, beliefs, and acceptability of human papillomavirus vaccination among parents in Chihuahua, Mexico

Introducción: La cobertura de vacunación contra el virus del papiloma humano no se ha realizado en la totalidad de la población, existen factores que interfieren en que los padres de las adolescentes acepten su aplicación. Objetivo: Relacionar el conocimiento sobre el virus del papiloma humano, el conocimiento sobre la vacuna contra el virus, las creencias sobre la vacuna con la aceptabilidad de la vacuna por los padres de las adolescentes de 9-12 años escolarizadas en Chihuahua, México.Materiales y Métodos: Estudio de tipo descriptivo, correlacional y transversal, la muestra fue de tipo censal, se conformó por 145 padres de niñas entre 9 a 12 años inscritas en tres primarias públicas ubicadas en una zona urbana de Chihuahua, México. Resultados: El conocimiento sobre el virus del papiloma humano se relacionó con la aceptabilidad de la vacuna (p < 0,009), de igual manera con el conocimiento acerca de la vacuna del virus del papiloma humano (p < 0,030) mientras que las creencias sobre el VPH y la vacuna no se relacionaron (p < 0, 747). Discusión: Los resultados coinciden con literatura previa en que el conocimiento sobre el virus y su vacuna es bajo, sin embargo, en este estudio las puntuaciones fueron más bajas. Mientras que la aceptabilidad de la vacuna contra el VPH tiende a ser alta al igual que estudios previos. Conclusiones: El conocimiento sobre el virus del papiloma humano y la vacuna se relacionaron con la aceptabilidad de los padres para aplicar la vacuna a sus hijas.

Introduction: Human papillomavirus vaccination coverage has not been achieved in the general population. There are factors that interfere with the acceptance of the vaccine by the parents of adolescent girls. Objective: To correlate knowledge of human papillomavirus, knowledge of the vaccine against the virus, and beliefs about the vaccine with vaccine acceptance among parents of adolescent girls aged 9-12 years in Chihuahua, Mexico. Materials and Methods: A descriptive, correlational, and cross-sectional study was conducted with a census sample of 145 parents of girls between the ages of 9 and 12 enrolled in three public elementary schools in an urban area of Chihuahua, Mexico. Results: Knowledge of human papillomavirus was related to vaccine acceptance (p < 0.009), as was knowledge of the human papillomavirus vaccine (p < 0.030). In contrast, beliefs about HPV and the vaccine were not related (p < 0.747). Discussion: The results are consistent with previous literature in that knowledge of the virus and its vaccine is low, but the scores were lower in this study. In contrast, HPV vaccine acceptance tends to be high, as in previous studies. Conclusions: Knowledge about human papillomavirus and the vaccine was associated with parental acceptance of giving it to their daughters.

Introdução: A cobertura vacinal contra o papilomavírus humano não tem sido realizada em toda a população, existem fatores que interferem na aceitação da sua aplicação pelos pais de meninas adolescentes. Objetivo: Relacionar o conhecimento sobre o papilomavírus humano, o conhecimento sobre a vacina contra o vírus, as crenças sobre a vacina com a aceitabilidade da vacina pelos pais de meninas adolescentes de 9 a 12 anos que frequentam a escola em Chihuahua, México. Materiais e Métodos: Estudo descritivo, correlacional e transversal, a amostra foi do tipo censitária, composta por 145 pais de meninas entre 9 e 12 anos matriculadas em três escolas primárias públicas localizadas em uma área urbana de Chihuahua, México. Resultados: O conhecimento sobre o papilomavírus humano esteve relacionado com a aceitabilidade da vacina (p < 0,009), da mesma forma com o conhecimento sobre a vacina contra o papilomavírus humano (p < 0,030), enquanto as crenças sobre o HPV e a vacina não foram relacionadas (p < 0,747). Discussão: Os resultados coincidem com a literatura anterior na medida em que o conhecimento sobre o vírus e sua vacina é baixo, porém, neste estudo as pontuações foram inferiores. Embora a aceitabilidade da vacina contra o HPV tenda a ser elevada como em estudos anteriores. Conclusões: O conhecimento sobre o papilomavírus humano e a vacina esteve relacionado à aceitabilidade dos pais em aplicar a vacina em suas filhas.

Probing the Biosafety of Implantable Artificial Secretory Granules for the Sustained Release of Bioactive Proteins.

Among bio-inspired protein materials, secretory protein microparticles are of clinical interest as self-contained, slow protein delivery platforms that mimic secretory granules of the human endocrine system, in which the protein is both the drug and the scaffold. Upon subcutaneous injection, their progressive disintegration results in the sustained release of the building block polypeptides, which reach the bloodstream for systemic distribution and subsequent biological effects. Such entities are easily fabricated in vitro by Zn-assisted cross-molecular coordination of histidine residues. Using cationic Zn for the assembly of selected pure protein species and in the absence of any heterologous holding material, these granules are expected to be nontoxic and therefore adequate for different clinical uses. However, such presumed biosafety has not been so far confirmed and the potential protein dosage threshold not probed yet. By selecting the receptor binding domain (RBD) from the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein as a model protein and using a mouse lab model, we have explored the toxicity of RBD-made secretory granules at increasing doses up to ∼100 mg/kg of animal weight. By monitoring body weight and biochemical blood markers and through the histological scrutiny of main tissues and organs, we have not observed systemic toxicity. Otherwise, the bioavailability of the material was demonstrated by the induction of specific antibody responses. The presented data confirm the intrinsic biosafety of artificial secretory granules made by recombinant proteins and prompt their further clinical development as self-contained and dynamic protein reservoirs.

Varroa Destructor Classification Using Legendre-Fourier Moments with Different Color Spaces.

Bees play a critical role in pollination and food production, so their preservation is essential, particularly highlighting the importance of detecting diseases in bees early. The Varroa destructor mite is the primary factor contributing to increased viral infections that can lead to hive mortality. This study presents an innovative method for identifying Varroa destructors in honey bees using multichannel Legendre-Fourier moments. The descriptors derived from this approach possess distinctive characteristics, such as rotation and scale invariance, and noise resistance, allowing the representation of digital images with minimal descriptors. This characteristic is advantageous when analyzing images of living organisms that are not in a static posture. The proposal evaluates the algorithm's efficiency using different color models, and to enhance its capacity, a subdivision of the VarroaDataset is used. This enhancement allows the algorithm to process additional information about the color and shape of the bee's legs, wings, eyes, and mouth. To demonstrate the advantages of our approach, we compare it with other deep learning methods, in semantic segmentation techniques, such as DeepLabV3, and object detection techniques, such as YOLOv5. The results suggest that our proposal offers a promising means for the early detection of the Varroa destructor mite, which could be an essential pillar in the preservation of bees and, therefore, in food production.

Conceptual DFT, machine learning and molecular docking as tools for predicting LD50 toxicity of organothiophosphates.

CONTEXT: Several descriptors from conceptual density functional theory (cDFT) and the quantum theory of atoms in molecules (QTAIM) were utilized in Random Forest (RF), LASSO, Ridge, Elastic Net (EN), and Support Vector Machines (SVM) methods to predict the toxicity (LD50) of sixty-two organothiophosphate compounds. The A-RF-G1 and A-RF-G2 models were obtained using the RF method, yielding statistically significant parameters with good performance, as indicated by R2 values for the training set (R2Train) and R2 values for the test set (R2Test), around 0.90. METHODS: The molecular structure of all organothiophosphates was optimized via the range-separated hybrid functional ωB97XD with the 6-311 + + G** basis set. Seven hundred and eighty-seven descriptors have been processed using a variety of machine learning algorithms: RF LASSO, Ridge, EN and SVM to generate a predictive model. The properties were obtained with Multiwfn, AIMALL and VMD programs. Docking simulations were performed by using AutoDock 4.2 and LigPlot + programs. All the calculations in this work are carried out in Gaussian 16 program package.

A polyphasic taxonomy analysis reveals the presence of an ecotype of Rahnella contaminans associated with the gut of Dendroctonus-bark beetles.

Species belonging to the genus Rahnella are dominant members of the core gut bacteriome of Dendroctonus-bark beetles, a group of insects that includes the most destructive agents of pine forest in North and Central America, and Eurasia. From 300 isolates recovered from the gut of these beetles, 10 were selected to describe an ecotype of Rahnella contaminans. The polyphasic approach conducted with these isolates included phenotypic characteristics, fatty acid analysis, 16S rRNA gene, multilocus sequence analyses (gyrB, rpoB, infB, and atpD genes), and complete genome sequencing of two isolates, ChDrAdgB13 and JaDmexAd06, representative of the studied set. Phenotypic characterization, chemotaxonomic analysis, phylogenetic analyses of the 16S rRNA gene, and multilocus sequence analysis showed that these isolates belonged to Rahnella contaminans. The G + C content of the genome of ChDrAdgB13 (52.8%) and JaDmexAd06 (52.9%) was similar to those from other Rahnella species. The ANI between ChdrAdgB13 and JaDmexAd06 and Rahnella species including R. contaminans, varied from 84.02 to 99.18%. The phylogenomic analysis showed that both strains integrated a consistent and well-defined cluster, together with R. contaminans. A noteworthy observation is the presence of peritrichous flagella and fimbriae in the strains ChDrAdgB13 and JaDmexAd06. The in silico analysis of genes encoding the flagellar system of these strains and Rahnella species showed the presence of flag-1 primary system encoding peritrichous flagella, as well as fimbriae genes from the families type 1, α, ß and σ mainly encoding chaperone/usher fimbriae and other uncharacterized families. All this evidence indicates that isolates from the gut of Dendroctonus-bark beetles are an ecotype of R. contaminans, which is dominant and persistent in all developmental stages of these bark beetles and one of the main members of their core gut bacteriome.

Diabetic neuropathy: Past, present, and future.

Background: A sedentary lifestyle and an unhealthy diet have considerably increased the incidence of diabetes mellitus worldwide in recent decades, which has generated a high rate of associated chronic complications. Methods: A narrative review was performed in MEDLINE, EMBASES and SciELO databases, including 162 articles. Results: Diabetic neuropathy (DN) is the most common of these complications, mainly producing two types of involvement: sensorimotor neuropathy, whose most common form is symmetric distal polyneuropathy, and autonomic neuropathies, affecting the cardiovascular, gastrointestinal, and urogenital system. Although hyperglycemia is the main metabolic alteration involved in its genesis, the presents of obesity, dyslipidemia, arterial hypertension, and smoking, play an additional role in its appearance. In the pathophysiology, three main phenomena stand out: oxidative stress, the formation of advanced glycosylation end-products, and microvasculature damage. Diagnosis is clinical, and it is recommended to use a 10 g monofilament and a 128 Hz tuning fork as screening tools. Glycemic control and non-pharmacological interventions constitute the mainstay of DN treatment, although there are currently investigations in antioxidant therapies, in addition to pain management. Conclusions: Diabetes mellitus causes damage to peripheral nerves, being the most common form of this, distal symmetric polyneuropathy. Control of glycemia and comorbidities contribute to prevent, postpone, and reduce its severity. Pharmacological interventions are intended to relieve pain.

Evaluation of the Hybrid Membrane of ZnO Particles Supported in Cellulose Acetate for the Removal of Lead.

Water polluted by discarded heavy metals such as lead is creating a global pollution problem. In this work, adsorption of Pb(II) was realized in batch studies by a hybrid membrane of cellulose acetate with ZnO particles. First, ZnO particles were prepared by precipitation and immobilized on the membrane. The hybrid membrane was elaborated by interfacial polymerization. The structure and surface were characterized based on Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). Batch experiments were carried out under different conditions where the number of particles of ZnO present in the membrane and the pH of the aqueous solution were varied. The Langmuir and Freundlich isotherm models were evaluated in the best adsorption conditions. Data fitted well with a Langmuir model with a maximum adsorption capacity of 15.55 mg·g-1, which was similar for this type of materials. Thermodynamic parameters such as Gibbs free energy, enthalpy, and entropy showed that the process was spontaneous and favorable. The hybrid membrane was evaluated in simulated wastewater of the battery industry with a superior efficiency of up to 97%; without the medium, it did not generate interference. These results suggest that Pb(II) removal by hybrid membrane is possible.

Escherichia coli as a New Platform for the Fast Production of Vault-like Nanoparticles: An Optimized Protocol.

Vaults are protein nanoparticles that are found in almost all eukaryotic cells but are absent in prokaryotic ones. Due to their properties (nanometric size, biodegradability, biocompatibility, and lack of immunogenicity), vaults show enormous potential as a bio-inspired, self-assembled drug-delivery system (DDS). Vault architecture is directed by self-assembly of the "major vault protein" (MVP), the main component of this nanoparticle. Recombinant expression (in different eukaryotic systems) of the MVP resulted in the formation of nanoparticles that were indistinguishable from native vaults. Nowadays, recombinant vaults for different applications are routinely produced in insect cells and purified by successive ultracentrifugations, which are both tedious and time-consuming strategies. To offer cost-efficient and faster protocols for nanoparticle production, we propose the production of vault-like nanoparticles in Escherichia coli cells, which are still one of the most widely used prokaryotic cell factories for recombinant protein production. The strategy proposed allowed for the spontaneous encapsulation of the engineered cargo protein within the self-assembled vault-like nanoparticles by simply mixing the clarified lysates of the producing cells. Combined with well-established affinity chromatography purification methods, our approach contains faster, cost-efficient procedures for biofabrication in a well-known microbial cell factory and the purification of "ready-to-use" loaded protein nanoparticles, thereby opening the way to faster and easier engineering and production of vault-based DDSs.

Metabarcoding of mycetangia from the Dendroctonus frontalis species complex (Curculionidae: Scolytinae) reveals diverse and functionally redundant fungal assemblages.

Dendroctonus-bark beetles are associated with microbes that can detoxify terpenes, degrade complex molecules, supplement and recycle nutrients, fix nitrogen, produce semiochemicals, and regulate ecological interactions between microbes. Females of some Dendroctonus species harbor microbes in specialized organs called mycetangia; yet little is known about the microbial diversity contained in these structures. Here, we use metabarcoding to characterize mycetangial fungi from beetle species in the Dendroctonus frontalis complex, and analyze variation in biodiversity of microbial assemblages between beetle species. Overall fungal diversity was represented by 4 phyla, 13 classes, 25 orders, 39 families, and 48 genera, including 33 filamentous fungi, and 15 yeasts. The most abundant genera were Entomocorticium, Candida, Ophiostoma-Sporothrix, Ogataea, Nakazawaea, Yamadazyma, Ceratocystiopsis, Grosmannia-Leptographium, Absidia, and Cyberlindnera. Analysis of α-diversity indicated that fungal assemblages of D. vitei showed the highest richness and diversity, whereas those associated with D. brevicomis and D. barberi had the lowest richness and diversity, respectively. Analysis of ß-diversity showed clear differentiation in the assemblages associated with D. adjunctus, D. barberi, and D. brevicomis, but not between closely related species, including D. frontalis and D. mesoamericanus and D. mexicanus and D. vitei. A core mycobiome was not statistically identified; however, the genus Ceratocystiopsis was shared among seven beetle species. Interpretation of a tanglegram suggests evolutionary congruence between fungal assemblages and species of the D. frontalis complex. The presence of different amplicon sequence variants (ASVs) of the same genus in assemblages from species of the D. frontalis complex outlines the complexity of molecular networks, with the most complex assemblages identified from D. vitei, D. mesoamericanus, D. adjunctus, and D. frontalis. Analysis of functional variation of fungal assemblages indicated multiple trophic groupings, symbiotroph/saprotroph guilds represented with the highest frequency (∼31% of identified genera). These findings improve our knowledge about the diversity of mycetangial communities in species of the D. frontalis complex and suggest that minimal apparently specific assemblages are maintained and regulated within mycetangia.

QSTR Modeling to Find Relevant DFT Descriptors Related to the Toxicity of Carbamates.

Compounds containing carbamate moieties and their derivatives can generate serious public health threats and environmental problems due their high potential toxicity. In this study, a quantitative structure-toxicity relationship (QSTR) model has been developed by using one hundred seventy-eight carbamate derivatives whose toxicities in rats (oral administration) have been evaluated. The QSRT model was rigorously validated by using either tested or untested compounds falling within the applicability domain of the model. A structure-based evaluation by docking from a series of carbamates with acetylcholinesterase (AChE) was carried out. The toxicity of carbamates was predicted using physicochemical, structural, and quantum molecular descriptors employing a DFT approach. A statistical treatment was developed; the QSRT model showed a determination coefficient (R2) and a leave-one-out coefficient (Q2LOO) of 0.6584 and 0.6289, respectively.

New QSPR model for prediction of corrosion inhibition using conceptual density functional theory.

The relationship between structure and corrosion inhibition of a series of twenty-eight quinoline and pyridine derivatives has been established through the investigation of quantum descriptors calculated with PBE/6-311 + + G** method. A quantitative structure-property relationship (QSPR) model was obtained by examining these descriptors using a genetic algorithm approximation method based on a multiple linear regression analysis. The results indicate that the efficiency of corrosion inhibitors is strongly associated with hardness (η), minimal electrostatic potential (ESPmin), and volume (V) descriptors. Furthermore, the validity of the proposed model is corroborated by an adsorption study on an iron surface Fe(110).

Rahnella sp., a Dominant Symbiont of the Core Gut Bacteriome of Dendroctonus Species, Has Metabolic Capacity to Degrade Xylan by Bifunctional Xylanase-Ferulic Acid Esterase.

Rahnella sp. ChDrAdgB13 is a dominant member of the gut bacterial core of species of the genus Dendroctonus, which is one of the most destructive pine forest bark beetles. The objectives of this study were identified in Rahnella sp. ChDrAdgB13 genome the glycosyl hydrolase families involved in carbohydrate metabolism and specifically, the genes that participate in xylan hydrolysis, to determine the functionality of a putative endo-1,4-ß-D-xylanase, which results to be bifunctional xylanase-ferulic acid esterase called R13 Fae and characterize it biochemically. The carbohydrate-active enzyme prediction revealed 25 glycoside hydrolases, 20 glycosyl transferases, carbohydrate esterases, two auxiliary activities, one polysaccharide lyase, and one carbohydrate-binding module (CBM). The R13 Fae predicted showed high identity to the putative esterases and glycosyl hydrolases from Rahnella species and some members of the Yersiniaceae family. The r13 fae gene encodes 393 amino acids (43.5 kDa), containing a signal peptide, esterase catalytic domain, and CBM48. The R13 Fae modeling showed a higher binding affinity to ferulic acid, α-naphthyl acetate, and arabinoxylan, and a low affinity to starch. The R13 Fae recombinant protein showed activity on α-naphthyl acetate and xylan, but not on starch. This enzyme showed mesophilic characteristics, displaying its optimal activity at pH 6.0 and 25°C. The enzyme was stable at pH from 4.5 to 9.0, retaining nearly 66-71% of its original activity. The half-life of the enzyme was 23 days at 25°C. The enzyme was stable in the presence of metallic ions, except for Hg2+. The products of R13 Fae mediated hydrolysis of beechwood xylan were xylobiose and xylose, manifesting an exo-activity. The results suggest that Rahnella sp. ChDrAdgB13 hydrolyze xylan and its products could be assimilated by its host and other gut microbes as a nutritional source, demonstrating their functional role in the bacterial-insect interaction contributing to their fitness, development, and survival.

All-in-one biofabrication and loading of recombinant vaults in human cells.

One of the most promising approaches in the drug delivery field is the use of naturally occurring self-assembling protein nanoparticles, such as virus-like particles, bacterial microcompartments or vault ribonucleoprotein particles as drug delivery systems (DDSs). Among them, eukaryotic vaults show a promising future due to their structural features,in vitrostability and non-immunogenicity. Recombinant vaults are routinely produced in insect cells and purified through several ultracentrifugations, both tedious and time-consuming processes. As an alternative, this work proposes a new approach and protocols for the production of recombinant vaults in human cells by transient gene expression of a His-tagged version of the major vault protein (MVP-H6), the development of new affinity-based purification processes for such recombinant vaults, and the all-in-one biofabrication and encapsulation of a cargo recombinant protein within such vaults by their co-expression in human cells. Protocols proposed here allow the easy and straightforward biofabrication and purification of engineered vaults loaded with virtually any INT-tagged cargo protein, in very short times, paving the way to faster and easier engineering and production of better and more efficient DDS.

Eukaryotic Aggresomes: Protocols and Tips for Their Production, Purification , and Handling.

Aggresomes are insoluble protein aggregates found in eukaryotic cells when the intracellular machinery is overtitered by, for example, the overexpression of a recombinant protein. These protein nanoparticles have become excellent models in studies devoted to elucidate protein aggregation processes in eukaryotic cells, like those involved in "conformational disorders" linked to neurodegenerative diseases. Since the presence of such protein aggregates is a hallmark of these conditions, they constitute an excellent target for new therapeutic approaches for such devastating pathologies. Moreover, and following the pathway opened a few years ago by bacterial inclusion bodies, eukaryotic aggresomes have been proposed as a new type of carrier-free, self-immobilized biocatalysts for use in biotechnology and biomedicine. Altogether, unraveling the characteristics and putative applications of naturally occurring protein aggregates has received an increasing interest during the last years. For that, availability of protocols allowing the production and purification of aggresomes constitute a valuable tool to boost research in the abovementioned fields. In this chapter, we describe both upstream and downstream protocols to obtain aggresomes produced in human cells, using as a model the recombinant human enzyme alpha-galactosidase A (GLA), together with technical tips and advices when working and analyzing eukaryotic aggresomes.

Revealing traces of depression through personal statements analysis in social media.

Depression is a common and very important health issue with serious effects in the daily life of people. Recently, several researchers have explored the analysis of user-generated data in social media to detect and diagnose signs of this mental disorder in individuals. In this regard, we tackled the depression detection task in social media considering the idea that terms located in phrases exposing personal statements (i.e., phrases characterized by the use of singular first person pronouns) have a special value for revealing signs of depression. First, we assessed the value of the personal statements for depression detection in social media. Second, we adapted an automatic approach that emphasizes the personal statements by means of a feature selection method and a term weighting scheme. Finally, we addressed the task in hand as an early detection problem, where the aim is to detect traces of depression with as much anticipation as possible. For evaluating these ideas, benchmark Reddit data for depression detection was used. The obtained results indicate that the personal statements have high relevance for revealing traces of depression. Furthermore, the results on early scenarios demonstrated that the proposed approach achieves high competitiveness compared with state-of-the-art methods, while maintaining its simplicity and interpretability.

Tolerability to non-endosomal, micron-scale cell penetration probed with magnetic particles.

The capability of HeLa cells to internalize large spherical microparticles has been evaluated by using inorganic, magnetic microparticles of 1 and 2.8 µm of diameter. In both absence but especially under the action of a magnet, both types of particles were uptaken, in absence of cytotoxicity, by a significant percentage of cells, in a non-endosomal process clearly favored by the magnetic field. The engulfed particles efficiently drive inside the cells chemically associated proteins such as GFP and human alpha-galactosidase A, without any apparent loss of protein functionalities. While 1 µm particles are completely engulfed, at least a fraction of 2.8 µm particles remain embedded into the cell membrane, with only a fraction of their surface in cytoplasmic contact. The detected tolerance to endosomal-independent cell penetration of microscale objects is not then restricted to organic, soft materials (such as bacterial inclusion bodies) as previously described, but it is a more general phenomenon also applicable to inorganic materials. In this scenario, the use of magnetic particles in combination with external magnetic fields can represent a significant improvement in the internalization efficiency of such agents optimized as drug carriers. This fact offers a wide potential in the design and engineering of novel particulate vehicles for therapeutic, diagnostic and theragnostic applications.

COVID-19 in Pregnancy: A Current Review of Global Cases.

IMPORTANCE: There is great concern about the impact of COVID-19 in pregnancy due to the high morbidity and mortality associated with prior coronavirus infections. OBJECTIVE: The objective of this review is to summarize the current literature on the impact of COVID-19 on pregnant women and their newborns. EVIDENCE ACQUISITION: The search terms COVID-19 and pregnancy were used in Medline and Clinical Key databases. Only articles written in English with outcome data on both mothers and their newborns were incorporated. RESULTS: Pregnant women generally experience COVID-19 as a mild-moderate illness. However, approximately 5% become critically ill. Women with underlying comorbidities seem more likely to experience severe morbidity. Newborns also generally have a favorable course. Vertical transmission in the intrauterine period is possible but rare. Infection control measures need to be taken to prevent transmission during the peripartum period. There is a paucity of data on infections in the first and second trimesters, but research from those infected in the third trimester indicates a possible link with preterm birth. There is a significant percentage of asymptomatic cases. Racial disparities are also being noted with disproportionate numbers of racial/ethnic minorities being affected. CONCLUSIONS: COVID-19 is generally experienced by pregnant women and their newborns as a mild to moderate illness, although a minority become critically ill and mortality does occur. This is more likely among those with underlying comorbidities, as in the general population. Asymptomatic cases heighten the need for increased testing and infection control measures. Racial disparities highlight underlying vulnerabilities and the need for increased research and policy changes.

Human α-Galactosidase A Mutants: Priceless Tools to Develop Novel Therapies for Fabry Disease.

Fabry disease (FD) is a lysosomal storage disease caused by mutations in the gene for the α-galactosidase A (GLA) enzyme. The absence of the enzyme or its activity results in the accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb3), in different tissues, leading to a wide range of clinical manifestations. More than 1000 natural variants have been described in the GLA gene, most of them affecting proper protein folding and enzymatic activity. Currently, FD is treated by enzyme replacement therapy (ERT) or pharmacological chaperone therapy (PCT). However, as both approaches show specific drawbacks, new strategies (such as new forms of ERT, organ/cell transplant, substrate reduction therapy, or gene therapy) are under extensive study. In this review, we summarize GLA mutants described so far and discuss their putative application for the development of novel drugs for the treatment of FD. Unfavorable mutants with lower activities and stabilities than wild-type enzymes could serve as tools for the development of new pharmacological chaperones. On the other hand, GLA mutants showing improved enzymatic activity have been identified and produced in vitro. Such mutants could overcome several complications associated with current ERT, as lower-dose infusions of these mutants could achieve a therapeutic effect equivalent to that of the wild-type enzyme.