Synthetic Methods for Azaheterocyclic Phosphonates and Their Biological Activity: An Update 2004-2024.

The increasing importance of azaheterocyclic phosphonates in the agrochemical, synthetic, and medicinal field has provoked an intense search in the development of synthetic routes for obtaining novel members of this family of compounds. This updated review covers methodologies established since 2004, focusing on the synthesis of azaheterocyclic phosphonates, of which the phosphonate moiety is directly substituted onto to the azaheterocyclic structure. Emphasizing recent advances, this review classifies newly developed synthetic approaches according to the ring size and providing information on biological activities whenever available. Furthermore, this review summarizes information on various methods for the formation of C-P bonds, examining sustainable approaches such as the Michaelis-Arbuzov reaction, the Michaelis-Becker reaction, the Pudovik reaction, the Hirao coupling, and the Kabachnik-Fields reaction. After analyzing the biological activities and applications of azaheterocyclic phosphonates investigated in recent years, a predominant focus on the evaluation of these compounds as anticancer agents is evident. Furthermore, emerging applications underline the versatility and potential of these compounds, highlighting the need for continued research on synthetic methods to expand this interesting family.

Facilitadores y barreras para el uso de la práctica basada en evidencia / Facilitators and barriers to the use of evidence-based practice / Facilitadores e barreiras ao uso de práticas baseadas em evidências

Objetivo: Conocer los facilitadores y barreras para el uso de la práctica basada en evidencia (PBE) en profesionales de enfermería que laboran en el área clínica. Método: Estudio cuali-tativo de tipo exploratorio descriptivo, con análisis de discurso, se realizaron 6 grupos focales conformados por 3 a 7 participantes para un total de 32 personas, se utilizó un muestreo inten-cional; en promedio fueron grabados entre 60 a 90 minutos por grupo focal. Para el análisis se empleó la fragmentación, codificación y categorización de las entrevistas obteniendo categorías temáticas, subcategorías y códigos a partir del uso del programa Atlas Ti versión 8. Resultados: El 69 % de los entrevistados respondió que el pregrado o posgrado habían recibido algún tipo de formación en PBE, la principal fuente de información cuando tienen alguna duda en la práctica clínica son los protocolos institucionales o los compañeros con más experiencia. Los facilitado-res identificados en este estudio fueron: tiempo para investigar, formación en PBE, iniciativa, grupos de investigación y compañeros; las barreras fueron: falta de apoyo institucional, poca formación en PBE, falta de tiempo, idioma, desmotivación profesional y el rechazo frente al cambio. Conclusiones: Se describen los facilitadores y barreras que presentan los profesionales de enfermería que laboran en el área clínica. Sin embargo se requieren investigaciones adicion-ales que permitan reconocer el fenómeno desde otras perspectivas en el contexto Colombiano y latinoamericano

Objective: To find out the facilitators and barriers to the use of evidence-based practice (EBP) in nursing professionals working in the clinical area. Method: Qualitative descrip-tive exploratory study, with discourse analysis, 6 focus groups were carried out with 3 to 7 participants for a total of 32, a purposive sampling was used; an average of 60 to 90 minutes were recorded per focus group. For the analysis we used the fragmentation, coding and cate-gorisation of the interviews obtaining thematic categories, subcategories and codes from the use of Atlas Ti version 8. Results: 69 % of the interviewees responded that the undergraduate or postgraduate had received some kind of training in EBP, the main source of information when they have any doubt in clinical practice are the institutional protocols or colleagues with more experience. Facilitators identified in this study were: time to investigate, training in EBP, initiative, research groups and peers; barriers were: lack of institutional support, little training in EBP, lack of time, language, professional demotivation and rejection in the face of change. Conclusions: The facilitators and barriers presented by nursing professionals working in the clinical area are described. However, further research is needed to recognise the phenomenon from other perspectives in the Colombian and Latin American context.

Objetivo: Descobrir os facilitadores e as barreiras ao uso da prática baseada em evidências (PBE) em profissionais de enfermagem que atuam na área clínica. Método: Estudo exploratório descritivo qualitativo, com análise de discurso; foram realizados 6 grupos focais com 3 a 7 participantes, totalizando 32; foi usada uma amostragem intencional; foi registrada uma média de 60 a 90 minutos por grupo focal. Para a análise utilizou-se a fragmentação, codificação e categorização das entrevistas obtendo-se categorias temáticas, subcategorias e códigos a partir da utilização do Atlas Ti versão 8. Resultados: 69% dos entrevistados responderam que a grad-uação ou pós-graduação havia recebido algum tipo de treinamento em PBE, a principal fonte de informação quando têm alguma dúvida na prática clínica são os protocolos institucionais ou colegas com mais experiência. Os facilitadores identificados neste estudo foram: tempo para in-vestigar, treinamento em PBE, iniciativa, grupos de pesquisa e colegas; as barreiras foram: falta de apoio institucional, pouco treinamento em PBE, falta de tempo, idioma, desmotivação profis-sional e rejeição diante de mudanças. Conclusões: São descritos os facilitadores e as barreiras apresentados pelos profissionais de enfermagem que trabalham na área clínica. No entanto, são necessárias mais pesquisas para reconhecer o fenômeno de outras perspectivas no contexto co-lombiano e latino-americano

Exploring COX-2 inhibitors in tuberculosis: A whole-blood model approach for immune response and adjunt therapy evaluation.

Pulmonary tuberculosis (TB) inflammation is an underestimated disease complication which anti-inflammatory drugs may alleviate. This study explored the potential use of the COX-2 inhibitors acetylsalicylic acid (ASA) and celecoxib in 12 TB patients and 12 healthy controls using a whole-blood ex vivo model where TNFα, PGE2, and LTB4 plasma levels were quantitated by ELISA; we also measured COX-2, 5-LOX, 12-LOX, and 15-LOX gene expression. We observed a significant TNFα production in response to stimulation with LPS or M. tuberculosis (Mtb). Celecoxib, but not ASA, reduced TNFα and PGE2 production, while increasing LTB4 in patients after infection with Mtb. Gene expression of COX-2 and 5-LOX was higher in controls, while 12-LOX was significantly higher in patients. 15-LOX expression was similar in both groups. We concluded that COX-2 inhibitors downregulate inflammation after Mtb infection, and our methodology offers a straightforward time-efficient approach for evaluating different drugs in this context. Further research is warranted to elucidate the underlying mechanisms and assess the potential clinical benefit.

Biomodulation of Induced Tooth Movement by Three Methods, Corticopuncture, Photobiomodulation, and Their Combination: An Animal Study.

Objective: The aim of this study was to assess the effects of accelerated tooth movement (ATM) methods: corticopuncture (CP), photobiomodulation (PBM), and their combination (CP + PBM) by evaluating tooth displacement, alveolar bone changes, and molecular and cellular response compared with conventional induced tooth movement. Materials and methods: Tooth movement and bone changes were evaluated on days 1, 3, and 7 (9 animals per time point) using microtomography, histological, and immunohistochemical evaluation, at compression and tension sites. CP groups received two perforations in the palate and one mesial to the molars. PBM was performed using GaAlAs diode laser applied every other day for 7 days (λ = 808 nm, 100 mW) in two points for 15 sec/point and total energy of 3 J. Results: Tooth movement was significantly increased in all three ATM groups after 7 days compared with the control group (mean 0.24 mm) by 27% PBM (0.31 mm), 45% CP (0.35 mm), and 57% CP + PBM (0.38 mm) (p < 0.05). At the compression side, all ATM groups showed significant decrease in bone density on day 3 (p < 0.05) and significant less bone volume on day 7 compared with Control (p < 0.05). At the tension side, PBM group showed a significant increase in bone density and volume on day 3 (p < 0.05). Immunohistochemistry analysis showed that at the compression side, tartrate-resistant acid phosphatase-positive cells, RANKL, and tumor necrosis factor-alpha expression were highly marked of the PBM and the combined method groups (p < 0.05). PBM and CP + PBM groups showed a significant increase in expression Runt-related transcription factor 2 and osterix (p < 0.05) at the tension side. Conclusions: All ATM groups showed increase on tooth displacement with CP + PBM group showing greatest tooth displacement. CP method appears to stimulate bone catabolism, PBM has more effect on bone formation, and the combined method showing a synergistic effect on bone remodeling.

Trastorno de depresión mayor e intento suicida en una paciente pediátrica víctima de abuso sexual / Major depressive disorder and suicidal attempt in a pediatric patient victim of sexual abuse

RESUMEN Introducción: el abuso sexual en la infancia se define como una experiencia traumática que puede llevar a la víctima a padecer complicaciones mentales, sexuales, físicas, interpersonales y comportamentales, nefastas para la salud. Además, constituye un factor de riesgo que desencadena el trastorno depresivo mayor en mujeres. Presentación del caso: se trata de una paciente femenina de 17 años de edad que sufrió abuso sexual a los siete años de edad. La paciente ingresa en una primera ocasión el 5 marzo de 2018 por intento suicida y trastorno depresivo al tener contacto con la persona que efectuó el acto. Pasados 17 días la paciente vuelve a ser ingresada por presentar un cuadro de trastorno depresivo mayor y egresa nueve días después por buena evolución. El 7 de agosto ingresa por tercera vez por trastorno depresivo mayor e intento suicida nuevamente. La paciente fue tratada con Imipramina de 25 mg dos veces al día y con tratamiento psicoterapéutico, al cual respondió adecuadamente. La paciente se encuentra psicológicamente estable. Conclusiones: el abuso sexual en la niñez constituye un factor de riesgo desencadenante del trastorno depresivo mayor. La atención especial mediante apoyo y estudios psicológicos de los pacientes, y el tratamiento de fármacos antidepresivos es de vital importancia para la recuperación de víctimas sometidas a acoso sexual con ideación e intento suicidas.

ABSTRACT Introduction: childhood sexual abuse is defined as a traumatic experience that can lead the victim to suffer mental, sexual, physical, interpersonal and behavioral complications harmful to health. It is also a risk factor for major depressive disorder in women. With this case, the authors aim to share experiences to the diagnosis; treatment and prevention of these health problems by disseminating them to the medical community. Case report: a 17-year-old female patient who was sexually abused at the age of 7 years. The patient is admitted on a first occasion on March 5, 2018 for suicide attempt and depressive disorder upon contact with the person who performed the act. After 17 days, the patient was admitted again for major depressive disorder and was discharged 9 days later due to good evolution. On August 7, she was admitted for a third time for major depressive disorder and another suicide attempt. The patient was treated with Imipramine 25 mg twice a day and psychotherapeutic treatment, responding satisfactorily. The patient is psychologically stable at present. Conclusions: sexual abuse in childhood constitutes a risk factor triggering major depressive disorder. Special attention through support and psychological studies of the patients and treatment with antidepressant drugs is of vital importance for the recovery of victims subjected to sexual harassment with suicidal ideation and attempt.

Sex-Dependent Differential Expression of Lipidic Mediators Associated with Inflammation Resolution in Patients with Pulmonary Tuberculosis.

There is a sex bias in tuberculosis's severity, prevalence, and pathogenesis, and the rates are higher in men. Immunological and physiological factors are fundamental contributors to the development of the disease, and sex-related factors could play an essential role in making women more resistant to severe forms of the disease. In this study, we evaluated sex-dependent differences in inflammatory markers. Serum samples were collected from 34 patients diagnosed with pulmonary TB (19 male and 15 female) and 27 healthy controls (18 male and 9 female). Cytokines IL2, IL4, IL6, IL8, IL10, IFNγ, TNFα, and GM-CSF, and eicosanoids PGE2, LTB4, RvD1, and Mar1 were measured using commercially available immunoassays. The MDA, a product of lipidic peroxidation, was measured by detecting thiobarbituric-acid-reactive substances (TBARS). Differential inflammation patterns between men and women were observed. Men had higher levels of IL6, IL8, and TNFα than women. PGE2 and LTB4 levels were higher in patients than healthy controls, but there were no differences for RvD1 and Mar1. Women had higher RvD1/PGE2 and RvD1/LTB4 ratios among patients. RvD1 plays a vital role in resolving the inflammatory process of TB in women. Men are the major contributors to the typical pro-inflammatory profile observed in the serum of tuberculosis patients.

High Vitamin D Concentrations Restore the Ability to Express LL37 by M. tuberculosis-Infected Human Macrophages.

Vitamin D has an immunomodulatory function and is involved in eliminating pathogens. Vitamin D deficiencies reported in Type 2 diabetes mellitus (T2DM) patients make them more susceptible to developing tuberculosis (TB). The macrophages are the immune cells that control intracellular pathogens by producing the antimicrobial peptide cathelicidin-LL37. This pathway involves TLR activation by pathogens, vitamin D receptor (VDR) ligation, and the enzyme 1α-hydroxylase Cytochrome P450 Family 27 Subfamily B Member 1 (CYP27B1). However, it is not clear whether the biological actions of vitamin D are affected by high glucose concentrations. This study aimed to evaluate the vitamin D contribution in the expression of VDR and CYP27B1, involved in the conversion of an inactive to an active form of vitamin D in the infected macrophages using M. tuberculosis as an infection model. The expression of LL37 and the nucleus translocation of VDR were evaluated as the readout of the response of vitamin D and determined if those processes are affected by glucose concentrations. Macrophages from healthy donors were cultured under glucose concentrations of 5.5, 15, or 30 mM, stimulated with vitamin D in inactive (25(OH)D3) or active (1,25(OH)2D3) forms, and infected with M. tuberculosis. The vitamin D-dependent induction of LL37 and the expression of VDR and CYP27B1 genes were analyzed by qPCR, and VDR translocation was analyzed in nuclear protein extracts by ELISA. M. tuberculosis downregulated the expression of LL37 regardless of the glucose concentration, whereas VDR and CYP27B1 upregulated it regardless of the glucose concentration. After evaluating two concentrations of vitamin D, 1 nM or 1 µM, the high concentration (1 µM) was necessary to restore the induction of LL37 expression in M. tuberculosis-infected macrophages. High concentrations of the inactive form of vitamin D restore the infected macrophages' ability to express LL37 regardless of the glucose concentration. This finding supports the idea that vitamin D administration in patients with T2DM could benefit TB control and prevention.

CD33 is downregulated by influenza virus H1N1pdm09 and induces ROS and the TNF-α, IL-1ß, and IL-6 cytokines in human mononuclear cells.

The influenza A virus (IAV) H1N1pdm09 induces exacerbated inflammation, contributing to disease complications. Inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), favor an inflammatory response that aids viral replication and survival. A pathway by which spontaneous TNF-α production occurs involves either the reduction of Siglec-3 (CD33) levels or the absence of its ligand, sialic acid. Influenza virus uses sialic acid to enter cells by reducing their expression; however, the role of CD33 in IAV H1N1pdm09 stimulation and its relationship with inflammation have not yet been studied. To evaluate the role of CD33 in proinflammatory cytokine production in IAV H1N1pdm09 stimulation, peripheral blood mononuclear cells from healthy subjects were incubated with IAV H1N1pdm09. We observed that the infection caused an increase in the mRNA expression of proinflammatory cytokines such as TNF-α, interleukin (IL)-1ß, and IL-6 and a significant reduction in CD33 expression by monocytes at an early stage of infection. Additionally, suppressor of cytokine signaling 3 (SOCS-3) mRNA expression was upregulated at 6 h, and reactive oxygen species (ROS) production increased at 1.5 h. Moreover, a significant reduction in CD33 expression on the cell surface of monocytes from influenza patients or of IAV H1N1pdm09-stimulated monocytes incubated in vitro was observed by flow cytometry. The results suggest that the decrease in CD33 and increase of SOCS-3 expression induced by IAV H1N1pdm09 triggered TNF-α secretion and ROS production, suggesting an additional way to exacerbate inflammation during viral infection.

Epidemiología de los pacientes hospitalizados con COVID-19 en un hospital de tercer nivel / Epidemiology of hospitalized patients with COVID-19 in a tertiary care hospital

Resumen Introducción: COVID-19, causada por el betacoronavirus SARS-CoV-2, ha saturado los sistemas de salud del mundo. Objetivo: Describir las características epidemiológicas de los pacientes atendidos en un hospital de tercer nivel. Métodos: Se realizó una cohorte retrospectiva de pacientes con diagnóstico o sospecha de COVID-19, del 23 de marzo al 31 de julio de 2020. Resultados: En el Hospital Central Militar se hospitalizaron 4401 pacientes, 35 % derechohabientes, 26 % civiles, 28 % militares en activo y solo 11 %, militares retirados. Predominó el sexo masculino, tanto en los pacientes hospitalizados como en los que fallecieron, el grupo O+ y la ausencia de comorbilidades; entre las comorbilidades que se observaron, las principales fueron el sobrepeso y la diabetes. La mediana de edad de los pacientes hospitalizados fue de 49 años, mientras que 62 años fue la edad de quienes fallecieron; las mujeres mayores de 51 años tuvieron mayor riesgo de fallecer. La tasa de letalidad ajustada fue de 18.5 %; 50 % falleció durante los primeros seis días. Conclusiones: En este estudio se lograron identificar las características epidemiológicas y se destacaron las principales comorbilidades en pacientes mexicanos con infección por SARS-CoV-2.

Abstract Introduction: COVID-19, caused by the betacoronavirus SARS-CoV-2, has overwhelmed the world's health systems. Objective: To describe the epidemiological characteristics of patients treated in a tertiary care hospital. Methods: A retrospective cohort study of patients diagnosed with or suspected of having COVID-19 from March 23 to July 31, 2020 was conducted. Results: 4,401 patients were hospitalized at the Central Military Hospital, out of which 35 % were beneficiaries, 26 % civilians, 28 % active military, and only 11%, retired military. Male gender predominated, both in hospitalized patients and in those who died, as well as the O+ group and absence of comorbidities; among the observed comorbidities, the main ones were overweight and diabetes. Hospitalized patients' median age was 49 years, while median age of those who died was 62 years; women older than 51 years had a higher risk of dying. Adjusted case fatality rate was 18.5 %; 50 % died within the first six days. Conclusions: In this study, the epidemiological characteristics and main comorbidities in Mexican patients with SARS-CoV-2 infection were identified.

Mycobacterium tuberculosis H37Rv Strain Increases the Frequency of CD3+TCR+ Macrophages and Affects Their Phenotype, but Not Their Migration Ability.

In mycobacterial infections, the number of cells from two newly discovered subpopulations of CD3+ myeloid cells are increased at the infection site; one type expresses the T cell receptor (CD3+TCRαß+) and the other does not (CD3+TCRαß-). The role of Mycobacterium tuberculosis (Mtb) virulence in generating these subpopulations and the ability of these cells to migrate remains unclear. In this study, monocyte-derived macrophages (MDMs) infected in vitro with either a virulent (H37Rv) or an avirulent (H37Ra) Mtb strain were phenotypically characterized based on three MDM phenotypes (CD3-, CD3+TCRαß+, and CD3+TCRαß-); then, their migration ability upon Mtb infection was evaluated. We found no differences in the frequency of CD3+ MDMs at 24 h of infection with either Mtb strain. However, H37Rv infection increased the frequency of CD3+TCRαß+ MDMs at a multiplicity of infection of 1 and altered the expression of CD1b, CD1c, and TNF on the surface of cells from both the CD3+ MDM subpopulations; it also modified the expression of CCR2, CXCR1, and CCR7, thus affecting CCL2 and IL-8 levels. Moreover, H37Rv infection decreased the migration ability of the CD3- MDMs, but not CD3+ MDMs. These results confirm that the CD3+ macrophage subpopulations express chemokine receptors that respond to chemoattractants, facilitating cell migration. Together, these data suggest that CD3+ MDMs are a functional subpopulation involved in the immune response against Mtb.

Introduction: Transition Metals and Sulfur.

The number of transition metal ions which are essential to life - also often called trace elements - increased steadily over the years. In parallel, the list of biological functions in which transition metals are involved, has grown, and is still growing tremendously. Significant progress has been made in understanding the chemistry operating at the biological sites where metal ions have been discovered. Early on, based on the application of physical, chemical, and biological techniques, it became likely that numerous of these metal centers carry sulfur ligands in their coordination sphere, such as sulfide (S2-), cysteine (RS-), or methionine (RSCH3). Notably, the structure and the reactivity of the metal active sites turned out to be quite different from anything previously observed in simple coordination complexes. Consequently, the prediction of active-site structures, based on known properties of transition metal ion complexes, turned out to be difficult and incorrect in many cases. Yet, biomimetic inorganic chemistry, via synthesis and detailed structural and electronic characterization of synthetic analogues, became an important factor and helped to understand the properties of the metal active sites. Striking advances came from molecular biology techniques and protein crystallography, as documented by the publication of the first high-resolution structures of iron-sulfur proteins and the blue copper protein plastocyanin approximately five decades ago. In this volume of METAL IONS IN LIFE SCIENCES the focus will be on some of the most intriguing, in our view, transition metal-sulfur sites discovered in living organisms. These include the type 1 Cu mononuclear center, the purple mixed-valent [Cu1.5+-(Cys)2-Cu1.5+] CuA, the tetranuclear copper-sulfide catalytic center of nitrous oxide reductase, the heme-thiolate site in cytochrome P450, the iron-sulfur proteins with bound inorganic (S2-) and organic (Cys-) sulfur, the pterin dithiolene cofactor (Moco) coordinated to either molybdenum or tungsten, the [8Fe-7S] P-cluster and the [Mo-7Fe-9S-C]-homocitrate catalytic site of nitrogenase, the siroheme-[4Fe-4S] center involved in the reduction of sulfite (SO32-) to hydrogen sulfide (H2S), the NiFeS sites of hydrogenases and CO dehydrogenase, and the zinc finger domains. We apologize to all researchers and their associates who have made tremendous contributions to our current knowledge of the steadily increasing transition metal sulfur sites in proteins and enzymes but are not mentioned here. These omissions are by no means intentional but merely the consequence of time and space. We are fully aware of the excellent books and authoritative reviews on various aspects of the subject, however, it is our motivation to cover in one single volume this exciting domain of bioinorganic chemistry.

Sulfur, the Versatile Non-metal.

The non-metallic chemical element sulfur, 3216S , referred to in Genesis as brimstone and identified as element by Lavoisier, is the tenth most abundant element in the universe and the fifth most common element on Earth. Important inorganic forms of sulfur in the biosphere are elemental sulfur (S8), sulfate (SO2-4), and sulfide (S2-), sulfite (SO2-3), thiosulfate, (S2O23), and polythionates (S3O62-; S4O62-). Because of its wide range of stable oxidation states, from +6to -2, sulfur plays important roles in central biochemistry as a structural and redoxactive element and is intimately related to life on Earth. Unusual reaction pathways involving sulfur compounds become possible by the specific properties of this element. Sulfur occurs in all the major classes of biomolecules, including enzymes, proteins, sugars, nucleic acids, vitamin cofactors, and metabolites. The flexibility of these biomolecules follows from its versatile chemistry. The best known sulfur mineral is perhaps pyrite (Fool's gold), with the chemical formula, FeS2. Sulfur radical anions, such as [S3].-, are responsible for the intense blue color of lapis lazuli, one of the most desired and expensive artists' materials. In the microbial world, inorganic sulfur compounds, e.g., elemental sulfur and sulfate, belong to the most important electron acceptors. Studies on microbial sulfur metabolism revealed many novel enzymes and pathways and advanced the understanding on metabolic processes used for energy conservation, not only of the microbes, but of biology in general. Transition metal sulfur complexes display intriguing catalytic activities, they provide surfaces and complex cavities in metalloenzymes that activate inert molecules such as H2, CO, N2 or N2O, and they catalyze the transformations of numerous organic molecules. Both thiamine diphosphate- (ThDP) and S-adenosyl- L-methionine- (SAM) dependent enzymes belong to Nature's most powerful catalysts with a remarkable spectrum of catalytic activities. In conclusion, given sulfur's diverse properties, evolution made an excellent choice in selecting sulfur as one the basic elements of life.

Cytochrome P450. The Dioxygen-Activating Heme Thiolate.

Cytochromes P450 (CYPs) are heme b-binding enzymes and belong to Nature's most versatile catalysts. They participate in countless essential life processes, and exist in all domains of life, Bacteria, Archaea, and Eukarya, and in viruses. CYPs attract the interest of researchers active in fields as diverse as biochemistry, chemistry, biophysics, molecular biology, pharmacology, and toxicology. CYPs fight chemicals such as drugs, poisonous compounds in plants, carcinogens formed during cooking, and environmental pollutants. They represent the first line of defense to detoxify and solubilize poisonous substances by modifying them with dioxygen. The heme iron is proximally coordinated by a thiolate residue, and this ligation state represents the active form of the enzyme. The Fe(III) center displays characteristic UV/Vis and EPR spectra (Soret maximum at 418 nm; g-values at 2.41, 2.26, 1.91). The Fe(II) state binds the inhibitor carbon monoxide (CO) to produce a Fe(II)-CO complex, with the major absorption maximum at 450 nm, hence, its name P450. CYPs are flexible proteins in order to allow a vast range of substrates to enter and products to leave. Two extreme forms exist: substrate-bound (closed) and substrate-free (open). CYPs share a sophisticated catalytic cycle that involves a series of consecutive transformations of the heme thiolate active site, with the strong oxidants compound I and II as key intermediates. Each of these high-valent Fe(IV) species has its characteristic features and chemical properties, crucial for the activation of dioxygen and cleavage of strong C-H bonds.

THP-1 cells increase TNF-α production upon LPS + soluble human IgG co-stimulation supporting evidence for TLR4 and Fcγ receptors crosstalk.

The lipopolysaccharide (LPS) of Gram-negative bacteria is recognized on human monocytes and macrophages by TLR4 and MD2 and induces the production of inflammatory cytokines; the LPS + IgG complexes co-stimulation increases the cytokine production, mediated by the Fc-γRIIa (CD32a). We stimulated human CD14 + monocytes or THP-1 cells with LPS or LPS + soluble human IgG (sIgG) and TNF-α transcription and production, assessed RT-qPCR, ELISA, or flow cytometry, was enhanced by 30% upon LPS + sIgG compared to LPS stimulation. LPS + sIgG co-stimulation affected the NF-κB pathway (p65 phosphorylation and nucleus translocation, and IkB- α degradation). The biochemical inhibition of IRAK 1/4 and Syk kinases suppressed the enhancer effect of LPS + sIgG on TNF- α production, suggesting the involvement of both MyD88 dependent and independent pathways. Our results suggest that during LPS activation, sIgG may participate in a TLR4 - Fc-γR crosstalk.

Expansin-like Exl1 from Pectobacterium is a virulence factor required for host infection, and induces a defence plant response involving ROS, and jasmonate, ethylene and salicylic acid signalling pathways in Arabidopsis thaliana.

Expansins are encoded by some phytopathogenic bacteria and evidence indicates that they act as virulence factors for host infection. Here we analysed the expression of exl1 by Pectobacterium brasiliense and Pectobacterium atrosepticum. In both, exl1 gene appears to be under quorum sensing control, and protein Exl1 can be observed in culture medium and during plant infection. Expression of exl1 correlates with pathogen virulence, where symptoms are reduced in a Δexl1 mutant strain of P. atrosepticum. As well as Δexl1 exhibiting less maceration of potato plants, fewer bacteria are observed at distance from the inoculation site. However, bacteria infiltrated into the plant tissue are as virulent as the wild type, suggesting that this is due to alterations in the initial invasion of the tissue. Additionally, swarming from colonies grown on MacConkey soft agar was delayed in the mutant in comparison to the wild type. We found that Exl1 acts on the plant tissue, probably by remodelling of a cell wall component or altering the barrier properties of the cell wall inducing a plant defence response, which results in the production of ROS and the induction of marker genes of the JA, ET and SA signalling pathways in Arabidopsis thaliana. Exl1 inactive mutants fail to trigger such responses. This defence response is protective against Pectobacterium brasiliense and Botrytis cinerea in more than one plant species.

Diagnósticos fonoaudiológicos y síntomas no motores en pacientes con Enfermedad de Parkinson / Phonoaudiological diagnoses and non-motor symptoms in patients with Parkinson's disease

Objetivo: Este estudio descriptivo se orientó a conocer el perfil clínico-epidemiológico de la Enfermedad de Parkinson (EP) y la coexistencia entre síntomas no motores (SNM) y diagnósticos fonoaudiológicos (DF). Método: La muestra estuvo conformada por 34 personas con Parkinson idiopático (26 hombres y 8 mujeres), cuyas historias clínicas fueron analizadas para describir la coexistencia de DF, como la hipofonía, la disprosodia, la disartria y la disfagia, con síntomas no motores, tales como: trastornos gastrointestinales, depresión, trastornos del sueño y deterioro cognitivo. Resultados: Los resultados señalan que las personas con Parkinson tenían edades entre los 25 a los 86 años. En cuanto a la fase, se clasificaron en: estadio I el 11,7%, II el 17,6%, III el 47%, IV el 14,7% y V el 8,8%. El 47% de los pacientes llegó al servicio de Fonoaudiología en una etapa avanzada de la EP. Los SNM más frecuentes fueron trastornos del sueño (67,6%), depresión (58,8%), alteraciones gastrointestinales (29,4%) y deterioro cognitivo (15%). Los DF se distribuyeron así: disprosodia (38%), hipofonía (33%), disartria (18%) y disfagia (11%). Discusión: se observa una alta frecuencia tanto de SNM (como la depresión y los trastornos del sueño), como de SF (especialmente disprosodia e hipofonía). Esta sintomatología provoca, por una parte, la reducción del deseo de relacionarse socialmente y por otro, dificultades para hacerse entender al presentar un volumen de voz reducido o prosodia (además de trastornos de la melodía, inflexiones, marcadores paralingüísticos) de la expresión oral del lenguaje. Conclusión: los trastornos del sueño y la depresión podrían tener un impacto negativo significativo en las funciones fonoaudiológico de las personas con Parkinson.

Objective: This descriptive study was aimed at understanding the clinical-epidemiological profile of Parkinson's disease (PD) and the coexistence between non-motor symptoms (NMS) and phonoaudiological diagnoses (PD). Methods: The sample comprised 34 people with idiopathic Parkinson's (26 men and 8 women). Their clinical histories were analysed to describe the coexistence of PD, such as hypophonia, dysprosody, dysarthria and dysphagia, with non-motor symptoms, such as gastrointestinal disorders, depression, sleep disorders and cognitive impairment. Results: The results indicate that people with Parkinson's are between the ages of 25 and 86. In terms of phase, they were classified as: stage I 11.7%, II 17.6%, III 47%, IV 14.7% and V 8.8%. 47% of patients reached the Speech Therapy service at an advanced stage of PD. The most frequent NMS were sleep disorders (67.6%), depression (58.8%), gastrointestinal disorders (29.4%) and cognitive impairment (15%). The PD were distributed as follows: dysprosody (38%), hypophonia (33%), dysarthria (18%) and dysphagia (11%). Discussion: a high frequency of both NMS (such as depression and sleep disorders) and PD (especially dysprosody and hypophonia) is observed. This symptomatology causes a reduction in the desire to relate socially, and difficulties in making oneself understood by presenting a reduced voice volume or prosody (in addition to melody of speech disorders, inflections, paralinguistic markers) of the oral language expression. Conclusion: sleep disorders and depression could have a significant negative impact on the speech and hearing functions of people with Parkinson's.

Expression of USP18 and IL2RA Is Increased in Individuals Receiving Latent Tuberculosis Treatment with Isoniazid.

BACKGROUND: The treatment of latent tuberculosis infection (LTBI) in individuals at risk of reactivation is essential for tuberculosis control. However, blood biomarkers associated with LTBI treatment have not been identified. METHODS: Blood samples from tuberculin skin test (TST) reactive individuals were collected before and after one and six months of isoniazid (INH) therapy. Peripheral mononuclear cells (PBMC) were isolated, and an in-house interferon-γ release assay (IGRA) was performed. Expression of chemokine ligand 4 (CCL4), chemokine ligand 10 (CXCL10), chemokine ligand 11 (CXCL11), interferon alpha (IFNA), radical S-adenosyl methionine domain-containing 2 (RSAD2), ubiquitin-specific peptidase 18 (USP18), interferon-induced protein 44 (IFI44), interferon-induced protein 44 like (IFI44L), interferon-induced protein tetratricopeptide repeats 1(IFIT1), and interleukin 2 receptor subunit alpha (IL2RA) mRNA levels were assessed by qPCR before, during, and after INH treatment. RESULTS: We observed significantly lower relative abundances of USP18, IFI44L, IFNA, and IL2RA transcripts in PBMC from IGRA-positive individuals compared to levels in IGRA-negative individuals before INH therapy. Also, relative abundance of CXCL11 was significantly lower in IGRA-positive than in IGRA-negative individuals before and after one month of INH therapy. However, the relative abundance of CCL4, CXCL10, and CXCL11 mRNA was significantly decreased and that of IL2RA and USP18 significantly increased after INH therapy, regardless of the IGRA result. Our results show that USP18, IFI44L, IFIT1, and IL2RA relative abundances increased significantly, meanwhile the relative abundance of CCL4, CXCL11, and IFNA decreased significantly after six months of INH therapy in TST-positive individuals. CONCLUSIONS: Changes in the profiles of USP18, IL2RA, IFNA, CCL4, and CXCL11 expressions during INH treatment in TST-positive individuals, regardless of IGRA status, are potential tools for monitoring latent tuberculosis treatment.

The Intracellular Growth of M. tuberculosis Is More Associated with High Glucose Levels Than with Impaired Responses of Monocytes from T2D Patients.

Diabetes mellitus, a metabolic disease characterized by hyperglycemia and poor glucose control, is a risk factor for Mycobacterium tuberculosis (M. tuberculosis) infection and the development of active tuberculosis. To evaluate whether M. tuberculosis infection susceptibility is associated with an intrinsic factor in monocytes from type 2 diabetes (T2D) patients or it is associated with hyperglycemia per se, we analyzed TLR-2 and TLR-4 expression by flow cytometry and the cytokines IL-1ß, IL-6, IL-8, IL-10, and TNF-α by cytometric bead array assays, either stimulated with TLR-2 and TLR-4 ligands or infected with M. tuberculosis in the whole blood from T2D patients (n = 43) and healthy subjects (n = 26) or in CD14+ monocytes from healthy subjects cultured in high glucose (HG) (30 mM). The intracellular growth of M. tuberculosis was evaluated by CFU counts at 0, 1, and 3 days in both monocytes from T2D patients and monocytes from healthy subjects cultured in HG. We did not find significant differences in TLR expression, cytokine production, or growth of M. tuberculosis in monocytes from T2D patients compared with those in monocytes from healthy subjects. Despite these results, in vitro assays of monocytes cultured with 30 mM glucose led to significantly increased TLR-2 and TLR-4 basal expression compared to those of monocytes cultured with 11 mM glucose (P < 0.05). Conversely, the production of IL-6 by TLR-2 ligand stimulation, of IL-1ß, IL-6, and IL-8 by TLR-4 ligand stimulation, and of IL-8 by M. tuberculosis infection significantly decreased in monocytes cultured in HG (P < 0.05). Additionally, the intracellular survival of M. tuberculosis increased in monocytes in HG after day 3 of culture (P < 0.05). In conclusion, HG decreased IL-8 production and the intracellular growth control of M. tuberculosis by monocytes, supporting the hypothesis that hyperglycemia plays an important role in the impaired immune responses to M. tuberculosis in patients with T2D.

Season and size of urban particulate matter differentially affect cytotoxicity and human immune responses to Mycobacterium tuberculosis.

Exposure to air pollution particulate matter (PM) and tuberculosis (TB) are two of the leading global public health challenges affecting low and middle income countries. An estimated 4.26 million premature deaths are attributable to household air pollution and an additional 4.1 million to outdoor air pollution annually. Mycobacterium tuberculosis (M.tb) infects a large proportion of the world's population with the risk for TB development increasing during immunosuppressing conditions. There is strong evidence that such immunosuppressive conditions develop during household air pollution exposure, which increases rates of TB development. Exposure to urban air pollution has been shown to alter the outcome of TB therapy. Here we examined whether in vitro exposure to urban air pollution PM alters human immune responses to M.tb. PM2.5 and PM10 (aerodynamic diameters <2.5µm, <10µm) were collected monthly from rainy, cold-dry and warm-dry seasons in Iztapalapa, a highly populated TB-endemic municipality of Mexico City with elevated outdoor air pollution levels. We evaluated the effects of seasonality and size of PM on cytotoxicity and antimycobacterial host immunity in human peripheral blood mononuclear cells (PBMC) from interferon gamma (IFN-γ) release assay (IGRA)+ and IGRA- healthy study subjects. PM10 from cold-dry and warm-dry seasons induced the highest cytotoxicity in PBMC. With the exception of PM2.5 from the cold-dry season, pre-exposure to all seasonal PM reduced M.tb phagocytosis by PBMC. Furthermore, M.tb-induced IFN-γ production was suppressed in PM2.5 and PM10-pre-exposed PBMC from IGRA+ subjects. This observation coincides with the reduced expression of M.tb-induced T-bet, a transcription factor regulating IFN-γ expression in T cells. Pre-exposure to PM10 compared to PM2.5 led to greater loss of M.tb growth control. Exposure to PM2.5 and PM10 collected in different seasons differentially impairs M.tb-induced human host immunity, suggesting biological mechanisms underlying altered M.tb infection and TB treatment outcomes during air pollution exposures.

Resolvin D1 (RvD1) and maresin 1 (Mar1) contribute to human macrophage control of M. tuberculosis infection while resolving inflammation.

Resolvins and protectins counter inflammation, enhance phagocytosis, induce bactericidal/permeability-increasing protein (BPI) expression, and restore inflamed tissue to homeostasis. Because modulating the inflammation/antiinflammation balance is important in Mycobacterium tuberculosis infection, we evaluated the effects of resolvins and protectins on human macrophages infected in vitro. Monocyte-derived macrophages were infected with M. tuberculosis H37Rv at a multiplicity of infection (MOI) of 5 and treated 1 h post-infection in vitro with 100 nM LXA4, RvD1, RvD2, PD1 or 150 nM Mar1. After 24 h, cytokine production was measured by Luminex, and BPI and cathelicidin LL37 expression was determined by real-time PCR. Macrophage bactericidal activity was assessed by colony-forming units (CFUs) 3 days posttreatment. Nuclear translocation of Nrf2 was assessed by ELISA, NFκB translocation was determined by imaging cytometry, and BPI production was determined by fluorescence microscopy. We found that all lipids reduced LPS-dependent and M. tuberculosis-induced TNF-α production. RvD1 and Mar1 also induced a significant reduction in M. tuberculosis intracellular growth. RvD1 and Mar1 elicited distinct immunomodulatory patterns. RvD1 induced upregulation of both antimicrobial effector genes (BPI and LL37) and cytokines (GM-CSF and IL-6). Mar1 induced only BPI overexpression. RvD1 and Mar1 induced NFκB nuclear translocation, but only Mar1 induced Nrf2 translocation. Inhibition of G protein-coupled receptor signaling in infected macrophages abrogated the regulatory effects of RvD1. In conclusion, RvD1 and Mar1 modulate the anti-inflammatory and antimicrobial properties of M. tuberculosis-infected human macrophages. Since both proresolving lipids are inducible and synthesized from dietary components, they have immunotherapeutic potential against tuberculosis when inflammation is uncontrolled.