Highly Sensitive Whole-Cell Mercury Biosensors for Environmental Monitoring.

Whole-cell biosensors could serve as eco-friendly and cost-effective alternatives for detecting potentially toxic bioavailable heavy metals in aquatic environments. However, they often fail to meet practical requirements due to an insufficient limit of detection (LOD) and high background noise. In this study, we designed a synthetic genetic circuit specifically tailored for detecting ionic mercury, which we applied to environmental samples collected from artisanal gold mining sites in Peru. We developed two distinct versions of the biosensor, each utilizing a different reporter protein: a fluorescent biosensor (Mer-RFP) and a colorimetric biosensor (Mer-Blue). Mer-RFP enabled real-time monitoring of the culture's response to mercury samples using a plate reader, whereas Mer-Blue was analysed for colour accumulation at the endpoint using a specially designed, low-cost camera setup for harvested cell pellets. Both biosensors exhibited negligible baseline expression of their respective reporter proteins and responded specifically to HgBr2 in pure water. Mer-RFP demonstrated a linear detection range from 1 nM to 1 µM, whereas Mer-Blue showed a linear range from 2 nM to 125 nM. Our biosensors successfully detected a high concentration of ionic mercury in the reaction bucket where artisanal miners produce a mercury-gold amalgam. However, they did not detect ionic mercury in the water from active mining ponds, indicating a concentration lower than 3.2 nM Hg2+-a result consistent with chemical analysis quantitation. Furthermore, we discuss the potential of Mer-Blue as a practical and affordable monitoring tool, highlighting its stability, reliance on simple visual colorimetry, and the possibility of sensitivity expansion to organic mercury.

Physioprophylaxis post-stress test in soccer players: effects blood lactate concentrations

Abstract Objective: To determine the effects of physioprophylaxis (PP) on blood lactate (BL) concentrations after maximal incremental stress test, considering that this is the application of techniques in sports physiotherapy to reduce signs of muscle fatigue that can trigger injuries due to overload. Materials and Methods: Quantitative study, experimental type, longitudinal section in 12 university players. The group is divided into one control group (CG) with recovery at rest without PP and another experimental group (EG) to which PP is applied at the end of the test. Blood lactate is recorded with Accutrend Plus at the beginning of the test, (1) five minutes after finishing the test (2) and after the PP (3) at two different moments for intra-subject analysis. Results: The following data were obtained regarding blood lactate clearance, Moment 1: Without Plan (WoP) 4.86±1.4 and With Plan (WP) 8.85±1.25 (p<0.05), moment 2: (WoP) 5.6±1.76 and (WP) 7.8±1.3 (p<0.05) in mmol/L, and intra-subject: (WoP): 5.25±1.58; (WP): 8.35±1.33 (p<0.05). Conclusions: The clearance of lactate in the blood at 30 minutes post stress test in the EG is bigger than the CG, because they recovered with the physioprophylactic plan.

Resumen Objetivo: Determinar los efectos de la fisioprofilaxis (PP) sobre las concentraciones de lactato en sangre (BL) tras la prueba de esfuerzo incremental máximo, considerando que se trata de la aplicación de técnicas en fisioterapia deportiva para reducir los signos de fatiga muscular que pueden desencadenar lesiones por sobrecarga. Materiales y métodos: Estudio cuantitativo, tipo experimental, sección longitudinal en 12 jugadores universitarios. El grupo se divide en un grupo control (GC) con recuperación en reposo sin PP y otro grupo experimental (GE) al que se aplica PP al final de la prueba. El lactato sanguíneo se registra con Accutrend Plus al inicio de la prueba, (1) cinco minutos después de finalizar la prueba (2) y después de la PP (3) en dos momentos diferentes para el análisis intra-sujeto. Resultados: Se obtuvieron los siguientes datos con respecto al aclaramiento de lactato en sangre, Momento 1: Sin Plan (WoP) 4.86 ± 1.4 y Con Plan (WP) 8.85 ± 1.25 (p <0.05), Momento 2: (WoP) 5.6 ± 1.76 y (WP) ) 7,8 ± 1,3 (p <0,05) en mmol / L, e intra-sujeto: (WoP): 5,25 ± 1,58; (WP): 8,35 ± 1,33 (p <0,05). Conclusiones: El aclaramiento de lactato en sangre a los 30 minutos post prueba de esfuerzo en el GE es mayor que en el GC, debido a que se recuperaron con el plan fisioprofiláctico.

SMARCE1 deficiency generates a targetable mSWI/SNF dependency in clear cell meningioma.

Mammalian SWI/SNF (mSWI/SNF) ATP-dependent chromatin remodeling complexes establish and maintain chromatin accessibility and gene expression, and are frequently perturbed in cancer. Clear cell meningioma (CCM), an aggressive tumor of the central nervous system, is uniformly driven by loss of SMARCE1, an integral subunit of the mSWI/SNF core. Here, we identify a structural role for SMARCE1 in selectively stabilizing the canonical BAF (cBAF) complex core-ATPase module interaction. In CCM, cBAF complexes fail to stabilize on chromatin, reducing enhancer accessibility, and residual core module components increase the formation of BRD9-containing non-canonical BAF (ncBAF) complexes. Combined attenuation of cBAF function and increased ncBAF complex activity generates the CCM-specific gene expression signature, which is distinct from that of NF2-mutated meningiomas. Importantly, SMARCE1-deficient cells exhibit heightened sensitivity to small-molecule inhibition of ncBAF complexes. These data inform the function of a previously elusive SWI/SNF subunit and suggest potential therapeutic approaches for intractable SMARCE1-deficient CCM tumors.

The FUS::DDIT3 fusion oncoprotein inhibits BAF complex targeting and activity in myxoid liposarcoma.

Mammalian SWI/SNF (mSWI/SNF or BAF) ATP-dependent chromatin remodeling complexes play critical roles in governing genomic architecture and gene expression and are frequently perturbed in human cancers. Transcription factors (TFs), including fusion oncoproteins, can bind to BAF complex surfaces to direct chromatin targeting and accessibility, often activating oncogenic gene loci. Here, we demonstrate that the FUS::DDIT3 fusion oncoprotein hallmark to myxoid liposarcoma (MLPS) inhibits BAF complex-mediated remodeling of adipogenic enhancer sites via sequestration of the adipogenic TF, CEBPB, from the genome. In mesenchymal stem cells, small-molecule inhibition of BAF complex ATPase activity attenuates adipogenesis via failure of BAF-mediated DNA accessibility and gene activation at CEBPB target sites. BAF chromatin occupancy and gene expression profiles of FUS::DDIT3-expressing cell lines and primary tumors exhibit similarity to SMARCB1-deficient tumor types. These data present a mechanism by which a fusion oncoprotein generates a BAF complex loss-of-function phenotype, independent of deleterious subunit mutations.

Biosupramolecular complexes of amphiphilic photosensitizers with human serum albumin and cucurbit[7]uril as carriers for photodynamic therapy.

In the present work, we evaluated the supramolecular interactions between three photosensitizers, namely toluidine blue O (TBO, positively charged) and two fatty acid conjugates of 6 and 14 carbon atoms chain lengths (TBOC6 and TBOC14), with human serum albumin (HSA) and the macrocycle cucurbit[7]uril (CB[7]), alone or in combination within a biosupramolecular system as potential carriers of photosensitizers for Photodynamic therapy (PDT). Binding studies were carried out using photophysical and calorimetric techniques and accompanied with molecular docking simulations. Amphiphilic photosensitizers, particularly TBOC14, showed stronger binding to HSA and (CB[7]). Comparing the different delivery systems, (CB[7]) had a marginal effect on cell uptake and phototoxicity in HeLa cells, while HSA showed enhanced cell uptake with phototoxicities that depended on the photosensitizer. Despite low cell uptake, the combination of both (CB[7]) and HSA was the most phototoxic, which illustrates the potential of combining these systems for PDT applications.

A review of shared decision-making: A call to arms for health care professionals.

OBJECTIVE: To illustrate the use of shared decision-making (SDM) and SDM tools and aids as the essential components in the care of asthma. DATA SOURCES: We reviewed individual randomized controlled studies conducted between 1998 and 2020 to compare SDM interventions and the use of SDM tools and aids for the care of asthma. All studies were published or translated in English. STUDY SELECTIONS: We excluded studies of interventions that involved multiple components other than the SDM intervention unless the control group also received these interventions. We evaluated the existing literature on both SDM tools and aids and the process of SDM to summarize in this review. RESULTS: Shared decision-making tools and aids most commonly clarify the diagnostics and options for a treatment. The 6 elements of SDM were clearly supported. We found no considerable association between the presence of these elements of SDM and asthma outcomes. CONCLUSION: We found that SDM for asthma and SDM tools and aids were often made to transfer information about asthma treatment options and their harms and benefits. The correlation between their support of SDM key elements and their impact on asthma outcomes is often difficult to ascertain but when present, there was positive correlation to improving risk communication, adherence, patient satisfaction, and possibly decreasing liability.

YejM Modulates Activity of the YciM/FtsH Protease Complex To Prevent Lethal Accumulation of Lipopolysaccharide.

Lipopolysaccharide (LPS) is an essential glycolipid present in the outer membrane (OM) of many Gram-negative bacteria. Balanced biosynthesis of LPS is critical for cell viability; too little LPS weakens the OM, while too much LPS is lethal. In Escherichia coli, this balance is maintained by the YciM/FtsH protease complex, which adjusts LPS levels by degrading the LPS biosynthesis enzyme LpxC. Here, we provide evidence that activity of the YciM/FtsH protease complex is inhibited by the essential protein YejM. Using strains in which LpxC activity is reduced, we show that yciM is epistatic to yejM, demonstrating that YejM acts upstream of YciM to prevent toxic overproduction of LPS. Previous studies have shown that this toxicity can be suppressed by deleting lpp, which codes for a highly abundant OM lipoprotein. It was assumed that deletion of lpp restores lipid balance by increasing the number of acyl chains available for glycerophospholipid biosynthesis. We show that this is not the case. Rather, our data suggest that preventing attachment of lpp to the peptidoglycan sacculus allows excess LPS to be shed in vesicles. We propose that this loss of OM material allows continued transport of LPS to the OM, thus preventing lethal accumulation of LPS within the inner membrane. Overall, our data justify the commitment of three essential inner membrane proteins to avoid toxic over- or underproduction of LPS.IMPORTANCE Gram-negative bacteria are encapsulated by an outer membrane (OM) that is impermeable to large and hydrophobic molecules. As such, these bacteria are intrinsically resistant to several clinically relevant antibiotics. To better understand how the OM is established or maintained, we sought to clarify the function of the essential protein YejM in Escherichia coli Here, we show that YejM inhibits activity of the YciM/FtsH protease complex, which regulates synthesis of the essential OM glycolipid lipopolysaccharide (LPS). Our data suggest that disrupting proper communication between LPS synthesis and transport to the OM leads to accumulation of LPS within the inner membrane (IM). The lethality associated with this event can be suppressed by increasing OM vesiculation. Our research has identified a completely novel signaling pathway that we propose coordinates LPS synthesis and transport.

Intestinal microbiota regulates tryptophan metabolism following oral infection with Toxoplasma gondii.

INTRODUCTION: The intestinal microbiota plays an important role in modulating host immune responses. Oral Toxoplasma gondii infection can promote intestinal inflammation in certain mice strains. The IDO-AhR axis may control tryptophan (Trp) metabolism constituting an important immune regulatory mechanism in inflammatory settings. AIMS: In the present study, we investigated the role of the intestinal microbiota on Trp metabolism during oral infection with T gondii. METHODS AND RESULTS: Mice were treated with antibiotics for four weeks and then infected with T gondii by gavage. Histopathology and immune responses were evaluated 8 days after infection. We found that depletion of intestinal microbiota by antibiotics contributed to resistance against T gondii infection and led to reduced expression of AhR on dendritic and Treg cells. Mice depleted of Gram-negative bacteria presented higher levels of systemic Trp, downregulation of AhR expression and increased resistance to infection whereas depletion of Gram-positive bacteria did not affect susceptibility or expression of AhR on immune cells. CONCLUSION: Our findings indicate that the intestinal microbiota can control Trp availability and provide a link between the AhR pathway and host-microbiota interaction in acute infection with T gondii.

Synthetic Approaches to Phosphasugars (2-oxo-1,2-oxaphosphacyclanes) Using the Anomeric Alkoxyl Radical ß-Fragmentation Reaction as the Key Step.

The anomeric alkoxyl radical ß-fragmentation (ARF) of carbohydrates possessing an electron-withdrawing group (EWG) at C2, promoted by PhI(OAc)2/I2, gives rise to an acyclic iodide through which a pentavalent atom of phosphorus can be introduced via the Arbuzov reaction. After selective hydrolysis and subsequent cyclization, the phosphonate or phosphinate intermediates can be converted into 2-deoxy-1-phosphahexopyranose and 2-deoxy-1-phosphapentopyranose sugars. The ARF of carbohydrates with an electron-donor group (EDG) at C2 proceeds by a radical-polar crossover mechanism, and the cyclization occurs by nucleophilic attack of a conveniently positioned phosphonate or phosphinate group to the transient oxocarbenium ion. This alternative methodology leads to 5-phosphasugars with a 4-deoxy-5-phosphapentopyranose framework. The structure and conformation of the 2-oxo-1,2-oxaphosphinane and 2-oxo-1,2-oxaphospholane ring systems in different carbohydrate models have been studied by NMR and X-ray crystallography.

Photochemical C-H Amination of Ethers and Geminal Difunctionalization Reactions in One Pot.

A mild, atom-economic, and metal-free α-C-H amination of ethers using relatively stable nonafluorobutanesulfonyl (nonaflyl, Nf) azide as the aminating reagent to give N-sulfonyl hemiaminals is reported. This enables unprecedented C(sp3 ) difunctionalization reactions, leading to diverse functionalized amino group containing compounds starting from simple ethers in one pot.

Pericytes Act as Key Players in Spinal Cord Injury.

Spinal cord injury results in locomotor impairment attributable to the formation of an inhibitory fibrous scar, which prevents axonal regeneration after trauma. The scarcity of knowledge about the molecular and cellular mechanisms involved in scar formation after spinal cord lesion impede the design of effective therapies. Recent studies, by using state-of-the-art technologies, including genetic tracking and blockage of pericytes in combination with optogenetics, reveal that pericyte blockage facilitates axonal regeneration and neuronal integration into the local neural circuitry. Strikingly, a pericyte subset is essential during scarring after spinal cord injury, and its arrest results in motor performance improvement. The arising knowledge from current research will contribute to novel approaches to develop therapies for spinal cord injury. We review novel advances in our understanding of pericyte biology in the spinal cord.

Individualised or Standardised Outcome Measures: A Co-habitation?

Mental health outcome measurement is conflicted between two different schools of thought which underlie the division between standardised (nomothetic) and individualised or patient-generated (idiographic) measures. The underpinning philosophies of both approaches have very different starting points in terms of how we understand the world. And yet the strengths of both may contribute something useful for patients and mental health services. We suggest a convergence of approaches with new thinking on options for co-habitation.

[Quality of life in chronic urticaria and its relationship with economic impact and disease control in patients attended to at the University Hospital of Monterrey, Mexico]. / Calidad de vida en urticaria crónica y su relación con el impacto económico y control de la enfermedad en pacientes atendidos en el Hospital Universitario de Monterrey, México.

BACKGROUND: Chronic urticaria (CU) manifests itself with hives and sometimes angioedema. Physical and social discomfort worsens patient quality of life. CU has an important impact on patients' economy. OBJECTIVE: To evaluate the relationship of quality of life (QoL) with economic burden and chronic urticaria control of in patients treated at our center. METHODS: Cross-sectional, descriptive, observational study. We included CU-diagnosed adult patients. The CU-Q2oL and UCT questionnaires in Spanish and the economic burden and comorbidities questionnaire developed by our center were applied. A sample size of 36 patients was calculated. RESULTS: 36 patients were included, out of which 58.3% were females. Mean age was 39.9 ± 15.6 years. Regarding QoL, 66.7% of patients reported being "A little" affected, 25% "Somewhat" and 8.3% "A lot", and its relationship with monthly income yielded a p-value of 0.017. 38.9% of patients reported having a monthly income of less than $ 5000 pesos. When disease control was compared with the CU-Q2oL, a significant difference was obtained for questions concerning itching. CONCLUSION: There was association between the quality of life of patients with chronic urticaria and monthly income, the lower the income, the more will the quality of life be affected. Furthermore, greater CU control was observed to reduce the negative effects on quality of life caused by itching.

Antecedentes: La urticaria crónica se manifiesta con erupciones y ocasionalmente con angioedema. La molestia física y social empeora la calidad de vida de los pacientes, quienes, además, enfrentan importantes gastos. Objetivo: Evaluar la relación de la calidad de vida con la carga económica y el control de la urticaria crónica. Métodos: Estudio transversal, descriptivo y observacional. Se incluyeron adultos con diagnóstico de urticaria crónica. Se aplicaron los cuestionarios CU-Q2oL y UCT en español y un cuestionario sobre carga económica y enfermedades coexistentes. Se calculó un tamaño de muestra de 36 pacientes. Resultados: Se incluyeron 36 pacientes, 58.3 % fue del sexo femenino. La edad fue de 39.9 años ± 15.6. Respecto a la calidad de vida, 66.7 % reportó poca afectación, 25 % bastante y 8.3 % mucha; la relación con el ingreso económico mensual obtuvo p = 0.017; 38.9 % refirió ingresos mensuales menores a $5000. Al comparar el control de la enfermedad con el CU-Q2oL se obtuvo diferencia significativa para las preguntas referentes a prurito. Conclusiones: Existió asociación entre calidad de vida de los pacientes con urticaria crónica e ingreso familiar mensual: a menor ingreso, más afectación de la calidad de vida. A mayor control de la enfermedad, menor afectación.

Role of Schwann cells in cutaneous wound healing.

Dermal wound healing is the process of repairing and remodeling skin following injury. Delayed or aberrant cutaneous healing poses a challenge for the health care system. The lack of detailed understanding of cellular and molecular mechanisms involved in this process hampers the development of effective targeted treatments. In a recent study, Parfejevs et al.-using state-of-the-art technologies, including in vivo sophisticated Cre/loxP techniques in combination with a mouse model of excisional cutaneous wounding-reveal that Schwann cells induce adult dermal wound healing. Strikingly, genetic ablation of Schwann cells delays wound contraction and closure, decreases myofibroblast formation, and impairs skin re-epithelization after injury. From a drug development perspective, Schwann cells are a new cellular candidate to be activated to accelerate skin healing. Here, we summarize and evaluate recent advances in the understanding of Schwann cells roles in the skin microenvironment.

Targeting glioblastoma-derived pericytes improves chemotherapeutic outcome.

Glioblastoma is the most common malignant brain cancer in adults, with poor prognosis. The blood-brain barrier limits the arrival of several promising anti-glioblastoma drugs, and restricts the design of efficient therapies. Recently, by using state-of-the-art technologies, including thymidine kinase targeting system in combination with glioblastoma xenograft mouse models, it was revealed that targeting glioblastoma-derived pericytes improves chemotherapy efficiency. Strikingly, ibrutinib treatment enhances chemotherapeutic effectiveness, by targeting pericytes, improving blood-brain barrier permeability, and prolonging survival. This study identifies glioblastoma-derived pericyte as a novel target in the brain tumor microenvironment during carcinogenesis. Here, we summarize and evaluate recent advances in the understanding of pericyte's role in the glioblastoma microenvironment.

Pericytes in the Premetastatic Niche.

The premetastatic niche formed by primary tumor-derived molecules contributes to fixation of cancer metastasis. The design of efficient therapies is limited by the current lack of knowledge about the details of cellular and molecular mechanisms involved in the premetastatic niche formation. Recently, the role of pericytes in the premetastatic niche formation and lung metastatic tropism was explored by using state-of-the-art techniques, including in vivo lineage-tracing and mice with pericyte-specific KLF4 deletion. Strikingly, genetic inactivation of KLF4 in pericytes inhibits pulmonary pericyte expansion and decreases metastasis in the lung. Here, we summarize and evaluate recent advances in the understanding of pericyte contribution to premetastatic niche formation. Cancer Res; 78(11); 2779-86. ©2018 AACR.

[Evaluation of vitamin D serum levels in asthmatic adults and their relationship with asthma control].

BACKGROUND: Decreased levels of vitamin D influence on the control and severity of asthma. OBJECTIVE: To evaluate the relationship between vitamin D serum levels and asthma control, as well as nutritional status, quality of life and patient comorbidities. METHODS: Cross-sectional, observational, descriptive study of 43 asthmatic patients older than 18 years of age. Multiple logistic and multiple linear regression multivariate analyses of variance were performed; a p-value ≤ 0.05 was considered to be statistically significant. RESULTS: Insufficient vitamin D levels were determined in 83.7 % of patients; 93 % had at least one asthma exacerbation in the previous year. There was no relationship between vitamin D serum levels and asthma control as measured by ACT and FEV1. There was a significant association between body mass index and vitamin D levels (p = 0.013). With a quality of life questionnaire for asthmatic adults, 76.7 % were recorded to have a poor quality of life. CONCLUSIONS: No relationship was observed between vitamin D serum levels and asthma control in the patients. Most patients had insufficient vitamin D serum levels, uncontrolled asthma, and poor quality of life. Overweight and grade I obesity were associated with vitamin D insufficient levels.

Antecedentes: Los niveles disminuidos de vitamina D influyen en el control y la gravedad del asma. Objetivo: Evaluar la relación entre niveles séricos de vitamina D y control del asma, así como estado nutricional, calidad de vida y comorbilidades del paciente. Métodos: Estudio transversal, observacional y descriptivo de 43 pacientes asmáticos mayores de 18 años. Se realizó análisis de varianza multivariados de regresión logística múltiple y lineal múltiples; se consideró estadísticamente significativa una p ≤ 0.05. Resultados: Se determinaron niveles insuficientes de vitamina D en 83.7 % de los pacientes; 93 % presentó al menos una exacerbación asmática en el último año. No se evidenció relación entre los niveles séricos de vitamina D y control del asma medido por el Asthma Control Test (ACT) y el volumen espiratorio forzado el primer segundo (FEV1). Se obtuvo asociación significativa entre el índice de masa corporal y niveles de vitamina D (p = 0.013). Con un cuestionario de calidad de vida en adultos asmáticos se registró que 76.7 % tenía mala calidad de vida. Conclusiones: No se observó relación entre los niveles séricos de vitamina D y el control del asma en los pacientes. La mayoría de los pacientes tenía niveles séricos de vitamina D insuficientes, asma descontrolada y mala calidad de vida. El sobrepeso y la obesidad grado I se asociaron con niveles insuficientes de vitamina D.

Removal of Aromatic Chlorinated Pesticides from Aqueous Solution Using ß-Cyclodextrin Polymers Decorated with Fe3O4 Nanoparticles.

This article describes the sorption properties of cyclodextrin polymers (nanosponges; NS) with the pesticides 4-chlorophenoxyacetic acid (4-CPA) and 2,3,4,6-tetrachlorophenol (TCF), including an evaluation of its efficiency and a comparison with other materials, such as granulated activated carbon (GAC). NS-pesticide complexes were characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray powder diffraction (XRPD), proton nuclear magnetic resonance (¹H-NMR), UV⁻VIS, and thermogravimetric analysis (TGA). This confirms the interactions of the guests with nanosponges and shows that the polymers have favorable sorption capacities for chlorinated aromatic guests. Our studies also show that the inclusion complex is predominantly favored for NS/CPA rather than those formed between TCF and NS due to the size of the adsorbate and steric effects. Sorption studies carried with repeated cycles demonstrate that NS polymers could be an improved technology for pollutant removal from aquatic environments, as they are very efficient and reusable materials. Our experiments and characterization by SEM, EDS, UV⁻VIS, and magnetization saturation (VSM) also show that NS is an optimal substrate for the deposition of magnetite nanoparticles, thus improving the usefulness and properties of the polymer, as the nanosponges could be retrieved from aqueous solution with a neodymium magnet without losing its efficiency as a pesticide sorbent.