Quantitative Shotgun Proteomic Analysis of Bacteria after Overexpression of Recombinant Spider Miniature Spidroin, MaSp1.

Spider silk has extraordinary mechanical properties, displaying high tensile strength, elasticity, and toughness. Given the high performance of natural fibers, one of the long-term goals of the silk community is to manufacture large-scale synthetic spider silk. This process requires vast quantities of recombinant proteins for wet-spinning applications. Attempts to synthesize large amounts of native size recombinant spidroins in diverse cell types have been unsuccessful. In these studies, we design and express recombinant miniature black widow MaSp1 spidroins in bacteria that incorporate the N-terminal and C-terminal domain (NTD and CTD), along with varying numbers of codon-optimized internal block repeats. Following spidroin overexpression, we perform quantitative analysis of the bacterial proteome to identify proteins associated with spidroin synthesis. Liquid chromatography with tandem mass spectrometry (LC MS/MS) reveals a list of molecular targets that are differentially expressed after enforced mini-spidroin production. This list included proteins involved in energy management, proteostasis, translation, cell wall biosynthesis, and oxidative stress. Taken together, the purpose of this study was to identify genes within the genome of Escherichia coli for molecular targeting to overcome bottlenecks that throttle spidroin overexpression in microorganisms.

Different Reaction Modes Operating in ansa-Half-Sandwich Magnesium Catalysts.

Magnesium-based catalysts are becoming popular for hydroelementation reactions specially using p-block reagents. Based on the seminal report from Schäfer's group (ChemCatChem 2022, 14, e202201007), our study demonstrates that the reaction mechanisms exhibit a far greater degree of complexity than originally presumed. Magnesium has a variety of coordination modes (and access to different hybridizations) which allows this electron-deficient centre to modulate its catalytic power depending on the σ-donor properties of the reagent. DFT calculations demonstrate several reaction channels closely operating in these versatile catalysts. In addition, variations in limiting energy barriers resulting from catalyst modifications were examined as a function of the Hammett constant, thereby predicting enhanced efficiency in reaction conversions.

Primary oral and sinonasal mucosal melanomas in Latin America: a systematic review.

Primary oral and sinonasal mucosal melanomas (POSNMMs) are aggressive neoplasms with limited therapeutic alternatives. The aim of this review was to characterize the demographic, clinical, immunohistochemical, and molecular information regarding these tumors in the Latin American population. Articles published in English, Spanish, or Portuguese (1990-2022) retrieved from the PubMed/MEDLINE, Scopus, CAS, Web of Science, EBSCO, and Google Academic databases were included. Thirty-three studies, with a total of 1212 cases, were identified. Clinicopathological data were available for 870 cases and immunohistochemical and/or molecular information for 342. Nineteen studies (57.6%) reported cases of oral melanoma, three (9.1%) sinonasal melanoma, and 11 (33.3%) oral and sinonasal melanoma. Fifteen studies (45.5%) provided only clinicopathological data, 12 (36.4%) reported only immunohistochemical data, two (6.1%) shared clinicopathological and immunohistochemical data, one (3.0%) offered clinicopathological, immunohistochemical, and molecular data, one (3.0%) provided immunohistochemical and molecular data, one (3.0%) clinicopathological and molecular data, and one (3.0%) only molecular data. The mean age of individuals with POSNMMs was 58 years, and slightly more were male (male 51.3%, female 48.7%). In Latin America, POSNMMs are a rare but aggressive malignancy with a poor prognosis and limited treatment options. Although molecular data and targeted therapy are still being researched, data from Latin America indicate the need for multicenter collaborative clinical trials to unite individual and isolated efforts.

Diversity of nitrogen-fixing and phosphorus-solubilizing bacteria associated with the rhizosphere of Andean maize in Ecuador.

A great diversity of microorganisms in the soil plays an important role in the sustainability of agricultural production systems. Among these microorganisms are bacteria that have the ability to fix atmospheric nitrogen or mineralize phosphorus, thus making it easily assimilable for plants. Maize is the main crop in the highlands of Ecuador (above 2000 meters) and it is predominantly traditional, using native seeds and very little or no agrochemicals. The National Institute of Agricultural Research (INIAP) has a collection of bacteria collected from the rhizosphere of maize in the highlands of Ecuador that has not been taxonomically identified. This research aimed to carry out a biochemical and genetic characterization to establish the identity of the collected nitrogen-fixing and phosphorus-solubilizing bacteria and to understand better the diversity of microorganisms present in the root biome of Andean maize. The hypothesis consisted of determining if there is a difference in the bacteria associated with the rhizosphere of maize in the Andean region of Ecuador compared with other regions. The bacteria underwent classical biochemical characterization based on catalase, oxidase, urease, sulfates, indole, sulfate-indole motility (SIM), and lactose, among others, and genetic identification by 16S rDNA ribosomal gene sequencing, PCR, and SANGER sequencing. A great diversity of microorganisms associated with the rhizosphere of the crop was found, including the genera Agrobacterium, Bacillus, Stenotrophomonas, Acinetobacter, Brevundimonas, Pseudomonas, and Pseudoxanthomonas. INIAP conserves these bacteria in a bank of microorganisms associated with crops of economic importance. They are useful for the development of biofertilizers that could contribute to a more sustainable agriculture in the region.

Short-pulsed Q-switched fiber laser using graphene oxide quantum dots based as saturable absorber.

In this work, we report the experimental study of a Q-switched optical fiber laser based on graphene oxide quantum dots (GOQDs) as saturable absorber (SA). GOQDs are fabricated by carbonization and exfoliation electrospun polyacrylonitrile (PAN) fibers. The results of Fourier Transform Infrared Spectroscopy (FTIR) showed bands caused by the CHs and C[bond, double bond]O groups associated with the GOQDs. The Raman spectrum showed the typical G and D bands of GOQDs. The size of the GOQDs, calculated by Transmission Electron Microscopy (TEM) was 6 nm; additionally, by high resolution TEM (HRTEM), an interplanar distance of 0.19 nm corresponding to the (002) direction of the graphene oxide was calculated. The SA was achieved using the photodeposition technique of the GOQDs onto the core of a single-mode optical fiber. The nonlinear characterization (NLC) of the GOQDs was carried out using the P-scan technique with a high-gain erbium-doped fiber amplifier (EDFA) at a wavelength of 1550 nm. The obtained results showed a saturable absorption behavior with a value of ß=-1.178x10-6(m/W) and a non-linear susceptibility of Im(χ(3))≈-1.573x10-7(esu). The experimental results of the SA, based on GOQDs as a switching device in a fiber laser, showed a typical behavior of a Q-switched laser by generating a pulsed emission at a wavelength of 1599 nm, a frequency from 2 to 16 kHz, and a maximum average output power of 1.3 mW.

Screening for chemicals with thyroid hormone-disrupting effects using zebrafish embryo.

Thyroid disruption is an increasingly recognized issue in the use and development of chemicals and new drugs, especially to help toxicologist to complement the reproductive and developmental toxicology information of chemicals. Still, adequate assessment methods are scarce and often suffer a trade-off between physiological relevance and labor- and cost-intensive assays. Here, we present a tiered approach for a medium-throughput screening of chemicals to identify their thyroid disrupting potential in zebrafish embryos as a New Approach Methodology (NAM). After identifying the maximum tolerated concentrations, we exposed zebrafish larvae to sub-adverse effect levels of the reference compounds benzophenone-2, bisphenol A, phenylthiourea, potassium perchlorate, propylthiouracil, and phloroglucinol to exclude any systemic toxicity. Applying the transgenic zebrafish line that carries a gene for the red fluorescence protein (Tg(tg:mCherry)) under the thyroglobulin promoter, we could identify the thyroid disrupting effects of the chemicals by a time and cost-effective image analysis measuring the fluorescence levels in the thyroid glands. Our observations could be confirmed by altered expression patterns of genes involved in the hypothalamus-pituitary-thyroid (HPT) axis. Finally, to anchor the observed thyroid disruption, we determined some changes in the Thyroid hormone levels of triiodothyronine (T3) and Thyroxine (T4) using a newly developed liquid chromatography mass spectrometric (LCMS) method. The presented approach carries the potential to extend the toolbox for legislative authorities and chemical producers for the assessment of thyroid-specific endocrine disruption and to overcome current challenges in the evaluation of endocrine disruptors.

Phase stabilization and optical response in the ultraviolet range, due to terbium concentration in hafnium oxide nanoparticles.

This work reports the influence of terbium trivalent ions on growing kinetics and the phase in hafnium oxide nanoparticles, as well as response to radiation in the range from 256 to 286. The nanoparticles were obtained by the hydrothermal route maintaining the temperature constant and varying the reaction time and the terbium concentration. The results show a gradual change in the phase with the concentration of terbium trivalent ions, going from the monoclinic phase, present at low concentrations, to the tetragonal phase, which appears from dopant concentrations greater than 5% at., increasing in amount with concentration of dopant. These phases appear as two perfectly defined morphologies. Furthermore, there is a significant change in radiative lifetimes when the tetragonal phase appears.

TL glow curve and kinetic analysis of Na2SiO3:Pr3+ under beta radiation effect.

Ionizing radiation dosimetry with thermoluminescence (TL) materials based on silicon or glass can be interesting in its potential use in radiation monitoring as the solution to the constant looking of development of new radiation detectors. In this work, TL characteristics of sodium silicate exposed to beta radiation effects were studied. TL response beta irradiated exhibited a glow curve with two peaks centered at 398 K and 473 K. Samples showed linearity from 0.55 to 13.2 Gy. TL readings after 10 times showed a repeatability with an error of less than 1%. Remain information showed significant losses during the first 24 h, but its information was almost constant after 72 h of storage. The Tmax-Tstop method exhibited three peaks which were mathematically analyzed with a general order deconvolution finding kinetic orders close to the second order for the first peak, meanwhile the kinetic order for the second peak and third peak are close to second order. Finally, the VHR method showed anomalous TL glow curve behavior with an increasing intensity TL as the heating rate increased.

Evaluation of the registry DIALYREG for the assessment of continuous renal replacement techniques in the critically ill patient.

Continuous renal replacement techniques (CRRT) can induce complications and monitoring is crucial to ensure patient safety. We designed a prospective multicenter observational and descriptive study using the DIALYREG registry, an online database located on a REDCap web-based platform that allows real-time data analysis. Our main objective was to identify CRRT-related complications in our intensive care units (ICUs) and implement security measures accordingly. From January 2019 to December 2020, we included 323 patients with admission diagnoses of medical illness (54%), sepsis (24%), postoperative care (20%), and trauma (2%). CRRT indications were homeostasis (42%), oliguria (26%), fluid overload (15%), and hemodynamic optimization (13%). The median initial therapy dose was 30 ml/kg/h (IQR 25-40), and dynamic adjustment was performed in 61% of the treatments. Sets were anticoagulated with heparin (40%), citrate (38%) or no anticoagulation (22%). Citrate anticoagulation had several advantages: more frequent dynamic CRRT dose adjustment (77% vs. 58% with heparin and 56% without anticoagulation, p < 0.05), longer duration of set (median of 55 h, IQR 24-72 vs. 23 h, IQR 12-48 with heparin and 12 h, IQR 12-31 without anticoagulation, p < 0.05), less clotting of the set (26% vs. 46.7% with heparin, p < 0.05), and lower incidence of hypophosphatemia (1% citrate vs. 6% with heparin and 5% without anticoagulation). It was also safe and effective in subgroup analysis of patients with liver disease or sepsis. The main global complications were hypothermia (16%), hypophosphatemia (13%) and metabolic acidosis (10%). Weaning of the therapy was achieved through early discontinuation (56%), nocturnal therapy transition (26%) and progressive SLED (18%). 52% of the patients were discharged from the hospital, while 43% died in the ICU and 5% died during hospitalization. We can conclude that the DIALYREG registry is a feasible tool for real-time control of CRRT in our ICU.

Posible influencia de factores no controlados en las concentraciones séricas de uracilo / Possible influence of uncontrolled factors on serum uracil concentrations

Antecedentes y objetivos: el déficit de dihidropirimidina deshidrogenasa (DPD) se ha asociado con un mayor riesgo de toxicidad tras exposición a fluoropirimidinas (FP). La determinación de las concentraciones plasmáticas de uracilo endógeno (U) es la prueba recomendada para identificar el déficit de DPD. Sin embargo, el valor de U puede verse afectado por diversos factores. El objetivo fue determinar la concentración sérica de U en una población candidata a recibir tratamiento con FP y comprobar si su distribución era compatible con la prevalencia del déficit parcial de DPD estimada en población caucásica. Material y métodos: estudio observacional prospectivo en el que se incluyeron pacientes oncológicos candidatos a tratamiento con FP. Para la determinación analítica se empleó un sistema Dionex Ultimate 3000 UHPLC, acoplado a un espectrómetro de masas cuadrupolo-orbitrap híbrido Q-exactive. Resultados: se incluyeron 77 pacientes con una edad media de 71 años. La media y la mediana de las concentraciones séricas de U fue 30,4 y 24,0 ng/ml, respectivamente. El rango fue de 7,1 a 139,7 ng/ml. Un 79,2% de los pacientes presentó un nivel de U comprendido entre 16 y 150 ng/ml, mostrando una diferencia estadísticamente significativa al compararlo con la prevalencia estimada en población caucásica (8%) (p-valor <0,0001). El método analítico empleado tiene un coeficiente de correlación R2 > 0,99 y un límite de detección <0,2 ng/ml. Conclusiones: es necesario llevar a cabo más estudios con un diseño dirigido a establecer las condiciones óptimas relativas al pretratamiento de las muestras a fin de evitar o minimizar la influencia de estos factores sobre los valores del analito. (AU)

Background and objective: dihydropyridine dehydrogenase (DPD) deficiency has been associated with an increased risk of toxicity after exposure to fluoropyrimidines (FP). Determination of endogenous uracil (U) plasma concentrations is the recommended test to identify DPD deficiency. However, the value of U can be affected by various factors. The objective was to determine the serum concentration of U in a population candidate to receive treatment with FP and to verify if its distribution was compatible with the prevalence of partial DPD deficiency estimated in the Caucasian population. Material and methods: prospective observational study in which cancer patients candidates for FP treatment were included. For the analytical determination, a Dionex Ultimate 3000 UHPLC system coupled to a Q-exactive hybrid quadrupole-orbitrap mass spectrometer was used. Results: 77 patients, with a mean age of 71 years, were included. The mean and median serum U concentrations were 30.4 and 24.0 ng/ml, respectively. The range was from 7.1 to 139.7 ng/ml. 79.2% of the patients presented a U level between 16 and 150ng/ml, showing a statistically significant difference when compared to the estimated prevalence in the Caucasian population (8%) (p-value <0.0001). The analytical method used has a correlation coefficient R2 > 0.99 and a detection limit <0.2 ng/ml. Conclusions: it is necessary to carry out more studies with a design aimed at establishing the optimal conditions related to the pretreatment of the samples in order to avoid or minimize the influence of these factors on the analyte values. (AU)

Thermally coupled energy levels of Eu 3+ within the BaHfO3 matrix, excited with UV radiation.

The present work reports the study of the thermally coupled energy levels of the trivalent europium ion in the barium hafnate matrix, in the temperature range of 289.7-323.8 K, through the application of ultraviolet radiation. Barium hafnate doped with 1, 3, and 5 atomic % trivalent europium ion was synthesized by the hydrothermal route, using chlorides as precursor elements. X-ray diffraction analysis indicates that the material crystallized in the perovskite cubic phase of barium hafnate, according to diffraction card file number #24-0102. Using high-resolution transmission electron microscopy, it was observed that the obtained crystals have an average size of 10 nm. From the photoluminescent analysis at room temperature, it was found that the sample doped with 3.0% atomic of trivalent europium displayed with the highest luminescent emission, while the photoluminescent analysis as a function of temperature showed that the bands associated with the 5D0 → 7FJ (J = 1,2,3,4) transitions have a strong dependence on temperature in the physical range.

Photocatalytic properties of Cu-containing ZnO nanoparticles and their antifungal activity against agriculture-pathogenic fungus.

In this work, nanoparticles (NPs) of ZnO, ZnO with Cu incorporated at 2 and 30 wt%, and CuO were prepared by the hydrothermal method. X-ray diffraction pattern (DRX) analysis showed that ZnO with high Cu incorporation (30 wt%) generates the formation of a composite oxide (ZnO/CuO), while X-ray photoelectron spectroscopy (XPS) of the Cu (2 wt%) sample indicated that Cu is incorporated as a dopant (ZnO/Cu2%). The samples with Cu incorporated had enhanced visible light absorption. Methyl orange (MO) dye was used to perform photocatalytic tests under UV radiation. The antifungal activity of the NPs was tested against four agricultural phytopathogenic fungi: Neofusicoccum arbuti, Alternaria alternata, Fusarium solani, and Colletotrichum gloeosporioides. The ZnO/Cu2% nanoparticles showed adequate photocatalytic and high antifungal activity in comparison to pure oxides and the composite sample.

GC-MS determination of malondialdehyde, acrolein, and 4-hydroxy-2-nonenal by ultrasound-assisted dispersive liquid-liquid microextraction in beverages.

A new, fast, simple, and effective ultrasound-assisted dispersive liquid-liquid microextraction procedure (UA-DLLME) for the gas chromatography-mass spectrometry (GC-MS) determination of malondialdehyde, acrolein, and 4-hydroxy-2-nonenal in beverages was successfully developed. 2,4-Dinitrophenylhydrazine derivatization was performed during extraction. An asymmetrical 3541//18 screening design and a central composite surface response design were used to investigate the influence of the most critical factors during the extraction process (ultrasound time and temperature, extraction and disperser solvents volumes, salt addition, and derivatization reagent concentration). According to FDA guidelines, the method was validated, achieving good linearities with r2 ≥ 0.9982, recoveries between 94.0 and 102.4%, and reproducibility with RSD lower than 4.5%. The method was applied to simultaneously determine the compounds in 60 different beverage samples, including beer, coffee, black tea, and fruit juices. The presence of secondary lipid oxidation products is demonstrated in beverages with a strong roasting process or oxidation.

Comportamiento de las colinesterasas tras condiciones de fatiga en corredores de fondo / Behaviour of cholinesterases after fatigue conditions in endurance runners

El objetivo del presente estudio fue evaluar el efecto de un entrenamiento intenso en atletas de resistencia sobre el comportamiento de las colinesterasas (ChE) tras condiciones de fatiga y su relación con otros marcadores de carga interna. Participaron 18 atletas de sexo masculino especialistas en pruebas de resistencia. Se evaluó las ChE y dos índices de variabilidad de la frecuencia cardiaca en tres momentos diferentes, previo al protocolo (BASAL), 15 minutos posterior al protocolo (FINAL) y 24 horas después del entrenamiento (24H). Un ANOVA de una vía con post-hoc de Tukey HSD se utilizó para comparar las medias. Se encontraron cambios significativos en las variables analizadas (p < .001) con tamaños de efecto muy grandes (d > 0.9) en los diferentes momentos y correlaciones moderadas entre variables (p < .001). El comportamiento de las ChE muestra un cambio significativo (p < .001) posterior al ejercicio y una relación con otros indicadores de carga interna. Nuestros resultados indican que las ChE tienen relación con la fatiga en el caso de los deportistas estudiados, pudiendo ser una medida para determinar la carga de entrenamiento. (AU)

The objective of the present study was to evaluate the effect of intense training in endurance athletes on the behaviour of cholinesterases (ChE) after fatigue conditions and its relationships with other internal load markers. 18 male athletes specializing in endurance events participated. ChEs and two index of heart rate variability were evaluated at three different moments, before the study protocol (BASAL), 15 minutes after (FINAL) and the day after finishing the training (24H). A one-way ANOVA with post-hoc Tukey HSD was used to compare means. Significant differences were found in the variables analysed (p < .001), with very large effect sizes (d > 0.9) between BASAL, FINAL and 24H and moderate correlations between ChE and LnRMSSD and SS (p < .001). The behaviour of the ChEs showed a significant change (p < .001) after exercise and relationship with other internal training load indicators. Our results indicate that ChEs are related with fatigue in the studied athletes and may be a measure for training load determination. (AU)

Seroprevalence of hepatitis B and C viruses in moderate and severe COVID-19 inpatients: A cross-sectional study at a referral center in Mexico.

INTRODUCTION AND OBJECTIVES: The emergence of SARS-CoV-2, which causes the coronavirus disease (COVID-19) has caused a great impact on healthcare systems worldwide, including hepatitis B and C viruses screening and elimination programs. The high number of COVID-19 hospitalizations represent a great opportunity to screen patients for hepatitis B virus (HBV) and hepatitis C virus (HCV), which was the aim of this study. MATERIAL AND METHODS: Cross-sectional, retrospective study performed between April 2020 and 20201 at a referral center in Mexico dedicated to the care of adults with severe/critical COVID-19. We retrieved clinical, demographic, and laboratory results from each patient´s medical records, including antibodies against HCV (anti-HCV), HBV surface antigen (HBsAg), antibodies against the HBV core antigen (anti-HBcAg), and antibodies against HBsAg (anti-HBsAg). RESULTS: Out of 3620 patients that were admitted to the hospital, 24 (0.66%), 4 (0.11%), and 72 (1.99%) tested positive for anti-HCV, HBsAg, and anti-HBcAg, respectively. Of all seronegative patients, 954 (27%) had undetectable anti-HBsAg and 401 (12%) had anti-HBsAg at protective levels. Blood transfusion was the most relevant risk factor. Only 9.7% of the anti-HBc positive, 25% of the HBsAg positive, and 52% of the anti-HCV positive were aware of their serological status. CONCLUSIONS: In this study we found a prevalence of anti-HCV of 0.66%, HBsAg in 0.11%, and isolated anti-HBcAg in 1.99%. We also found that HBV vaccination coverage has been suboptimal and needs to be reinforced. This study gave us a trustworthy insight of the actual seroprevalence in Mexico, which can help provide feedback to the Hepatitis National Elimination Plan.

Epicardial fat thickness in infants of diabetic mothers.

BACKGROUND: Epicardial fat thickness (EFT) in adults and children has been related to the metabolic syndrome and other cardiovascular risk factors; however, scarce studies have evaluated it in infants of diabetic mothers (IDM) in whom, alterations in the thickness of the interventricular septum have been reported. This study compares the EFT in IDM versus infants of non-diabetic mothers (INDM) and its association with others echocardiographic parameters. METHODS: We performed a cross sectional study in 93 infants (64 IDM and 29 INDM). To evaluate EFT dimensions, an echocardiogram was performed within the first 24h of extrauterine life in both groups. In diabetic mothers, HbA1c was also determined. RESULTS: There was no significant difference in birth weight between the groups although gestational age was lower in IDM. The EFT (3.6 vs. 2.5 mm, p < 0.0001), the interventricular septum thickness (IVST) (6.2 vs. 5.2 mm, p < 0.0001) and the IVST / left ventricle posterior wall (1.3 vs. 1.1, p = 0.001) were higher in the IDM; while the left ventricular expulsion fraction [LVFE] (71.1 vs. 77.8; p < 0.0001) was lower than in the INDM, respectively. We found a positive correlation between EFT with IVST (r = 0.577; p = 0.0001), LVPW (r = 0.262; p = 0.011), IVST/LVPW index (r = 0.353; p = 0.001), and mitral integral early velocity (r = 0.313; p = 0.002), while a negative correlation with LVFE was observed (r = -0.376; p = 0.0001). CONCLUSIONS: The EFT is higher in IDM than in INDM. It was positively related with echocardiographic parameters of left ventricular thickness and negatively with left ventricular ejection function.

3D trapping of microbubbles by the Marangoni force.

In this Letter, we show 3D steady-state trapping and manipulation of vapor bubbles in liquids employing a low-power continuous-wave laser using the Marangoni effect. Light absorption from photodeposited silver nanoparticles on the distal end of a multi-mode optical fiber is used to produce bubbles of different diameters. The thermal effects produced by either the nanoparticles on the fiber tip or the light bulk absorption modulate the surface tension of the bubble wall and creates both longitudinal and transversal forces just like optical forces, effectively creating a 3D potential well. Using numerical simulations, we obtain expressions for the temperature profiles and present analytical expressions for the Marangoni force. In addition, using an array of three fibers with photodeposited nanoparticles is used to demonstrate the transfer of bubbles from one fiber to another by sequentially switching on and off the lasers.