Synergistic Experimental and Computational Investigation of the Bioorthogonal Reactivity of Substituted Aryltetrazines.

Tetrazines (Tz) have been applied as bioorthogonal agents for various biomedical applications, including pretargeted imaging approaches. In radioimmunoimaging, pretargeting increases the target-to-background ratio while simultaneously reducing the radiation burden. We have recently reported a strategy to directly 18F-label highly reactive tetrazines based on a 3-(3-fluorophenyl)-Tz core structure. Herein, we report a kinetic study on this versatile scaffold. A library of 40 different tetrazines was prepared, fully characterized, and investigated with an emphasis on second-order rate constants for the reaction with trans-cyclooctene (TCO). Our results reveal the effects of various substitution patterns and moreover demonstrate the importance of measuring reactivities in the solvent of interest, as click rates in different solvents do not necessarily correlate well. In particular, we report that tetrazines modified in the 2-position of the phenyl substituent show high intrinsic reactivity toward TCO, which is diminished in aqueous systems by unfavorable solvent effects. The obtained results enable the prediction of the bioorthogonal reactivity and thereby facilitate the development of the next generation of substituted aryltetrazines for in vivo applications.

Development of 18F-Labeled Bispyridyl Tetrazines for In Vivo Pretargeted PET Imaging.

Pretargeted PET imaging is an emerging and fast-developing method to monitor immuno-oncology strategies. Currently, tetrazine ligation is considered the most promising bioorthogonal reaction for pretargeting in vivo. Recently, we have developed a method to 18F-label ultrareactive tetrazines by copper-mediated fluorinations. However, bispyridyl tetrazines-one of the most promising structures for in vivo pretargeted applications-were inaccessible using this strategy. We believed that our successful efforts to 18F-label H-tetrazines using low basic labeling conditions could also be used to label bispyridyl tetrazines via aliphatic nucleophilic substitution. Here, we report the first direct 18F-labeling of bispyridyl tetrazines, their optimization for in vivo use, as well as their successful application in pretargeted PET imaging. This strategy resulted in the design of [18F]45, which could be labeled in a satisfactorily radiochemical yield (RCY = 16%), molar activity (Am = 57 GBq/µmol), and high radiochemical purity (RCP > 98%). The [18F]45 displayed a target-to-background ratio comparable to previously successfully applied tracers for pretargeted imaging. This study showed that bispyridyl tetrazines can be developed into pretargeted imaging agents. These structures allow an easy chemical modification of 18F-labeled tetrazines, paving the road toward highly functionalized pretargeting tools. Moreover, bispyridyl tetrazines led to near-instant drug release of iTCO-tetrazine-based 'click-to-release' reactions. Consequently, 18F-labeled bispyridyl tetrazines bear the possibility to quantify such release in vivo in the future.

Novel Mechanisms Targeted by Drug Trials in Pulmonary Arterial Hypertension.

Pulmonary arterial hypertension (PAH) is a rare disease associated with abnormally elevated pulmonary pressures and right heart failure resulting in high morbidity and mortality. Although the prognosis for patients with PAH has improved with the introduction of pulmonary vasodilators, disease progression remains a major problem. Given that available therapies are inadequate for preventing small-vessel loss and obstruction, there is active interest in identifying drugs capable of targeting angiogenesis and mechanisms involved in the regulation of cell growth and fibrosis. Among the mechanisms linked to PAH pathogenesis, preclinical studies have identified promising compounds that are currently being tested in clinical trials. These drugs target seven of the major mechanisms associated with PAH pathogenesis: bone morphogenetic protein signaling, tyrosine kinase receptors, estrogen metabolism, extracellular matrix, angiogenesis, epigenetics, and serotonin metabolism. In this review, we discuss the preclinical studies that led to prioritization of these mechanisms, and discuss completed and ongoing phase 2/3 trials using novel interventions such as sotatercept, anastrozole, rodatristat ethyl, tyrosine kinase inhibitors, and endothelial progenitor cells, among others. We anticipate that the next generation of compounds will build on the success of the current standard of care and improve clinical outcomes and quality of life for patients with PAH.

Prevalence and importance of non-tuberculous mycobacteria in adult patients with cystic fibrosis in a hospital in Madrid. / Prevalencia e importancia de micobacterias no tuberculosas en pacientes adultos con fibrosis quística en un hospital de Madrid.

INTRODUCTION: The role of non-tuberculous mycobacteria (NTM) among cystic fibrosis (CF) patients, on occasion, remains unknown. The aim of our study is to evaluate the prevalence and clinical/microbiological characteristics of CF adult patients colonized by NTM, highlighting Mycobacterium abscessus (M. abscessus). METHODS: A retrospective study was conducted with 92 CF adult patients: including a control group of 64 patients, not colonized by NTM, and a study group of 28 patients, colonized by NTM. We have analyzed variables such as age, F508del mutation, lung function, pancreatic involvement, auramine staining and co-colonizations between both groups. RESULTS: The prevalence of NTM found was 30.4%. The most prevalent was Mycobacterium avium complex followed by M. abscessus. For M. abscessus, in the comparative study with patients colonized by other NTM, significant results were obtained for variables age. DISCUSSION: We have found a high prevalence of NTM among adult patients with CF, and we associated the presence of M. asbcessus with ages less than 30 years and F508del. Due to the pathogenic role of NTM, especially M. asbcessus, multicenter studies are required within the population suffering from CF.

Atypical Association of Angelman Syndrome and Klinefelter Syndrome in a Boy with 47,XXY Karyotype and Deletion 15q11.2-q13.

Angelman syndrome (AS, OMIM 105830) is a neurogenetic disorder with firm clinical diagnostic guidelines, characterized by severe developmental delay and speech impairment, balanced and behavioral disturbance as well as microcephaly, seizures, and a characteristic electroencephalogram (EEG). The majority of AS cases (70%) are caused by a 15q11.2-q13 deletion on the maternally derived chromosome. The frequency of AS has been estimated to be between 1/10000 and 1/20000. Klinefelter syndrome (KS) occurs due to the presence of an extra X chromosome (karyotype 47,XXY). The main features in KS are small testes, hypergonadotropic hypogonadism, gynecomastia, learning difficulties, and infertility. We present what is, to our knowledge, the first case of a patient with both KS and AS due to a 15q11.2-q13 deletion on the maternally derived chromosome and an extra X chromosome of paternal origin. He showed dysmorphic features, axial hypotonia, and delayed acquisition of motor skills. Early diagnosis is essential for optimal treatment of AS children; this is one of the earliest diagnosed cases of AS probably due to the presence of two syndromes. Clinical findings in this patient here described may be helpful to identify any other cases and to evaluate recurrence risks in these families.

First report of a variant bovine papillomavirus type 2 (BPV-2) in cattle in the Iberian Peninsula.

Infections caused by bovine papillomavirus (BPV) have been described worldwide. Some types, like BPV-1 and BPV-2, have been reported in association with skin warts and fibropapillomas in cattle and sarcoids in equids. In this study we have investigated the presence of BPV in cutaneous warts isolated from a steer in Spain. Cutaneous fibropapillomatosis was confirmed by histopathological analysis. Complete genome was amplified by multiple-primed rolling circle and the L1, E5 and E6 genes were sequenced. The isolate was classified as a variant of BPV-2 on the basis of the L1 gene sequences. Genetic variability of L1, E5 and E6 genes was compared with BPV-2 isolates from different hosts in several continents. Some mutations involved non-synonymous substitutions when compared to the prototype strain. One of these non-conservative mutations was located in the jelly roll ß-barrel of the EF loop of the capsid protein (encoded by L1). This study presents the first report of a variant of BPV-2 infection in the Iberian Peninsula and contributes to extend the knowledge of the spreading and circulation of BPV.

Papel de la abuela en la lactancia materna / The Grandmother's Role in Breastfeeding / O papel da avó na lactância materna

Objetivo: investigar la influencia de la abuela sobre la madre que da lactancia materna y su relación con la incidencia y prevalencia de la misma. Método: se realizó una revisión integral consultando de las bases de datos MEDLINE OVID, DIALNET y COCHRANEC LIBRARY. Tras el análisis de los textos, se escogieron once estudios de investigación que iban dirigidos directamente a comprobar la influencia de las abuelas en la incidencia y prevalencia de la lactancia materna en diferentes contextos. Resultados: la abuela apareció como fuente de apoyo en población rural (41%). En población urbana la abuela aparece entre los tres factores principales (intención de la madre de lactar o no, decisión de la madre del patrón de alimentación de su hijo e información objetiva sobre pautas de amamantamiento), tanto en el papel de apoyo (94,7%), como en el de obstáculo (26,7%). En cuanto a la decisión de la madre sobre el patrón de alimentación de sus hijos, el 78% refiere haber tomado la decisión antes del embarazo o en el primer trimestre. La convivencia con la abuela tuvo asociación positiva con dar agua o infusiones (PR regresión de Cox con IC 95%: 1,51 [1,10-2,09]; p = 0,011). Conclusiones: a la luz de la revisión, la influencia detractora o no que ejerce la abuela sobre la lactancia materna la da sobre los tres factores principales. La influencia detractora ejercida por los consejos de la abuela parece estar relacionada con conocimientos erróneos adquiridos como "generación del biberón" más que con la intencionalidad, y es susceptible de modificarse con información adecuada, con resultados favorables sobre el patrón de alimentación.

Objective: to investigate the effect the grandmother has on the breastfeeding mother and how it relates to the incidence and prevalence of breastfeeding. Materials and methods: A comprehensive database review was conducted in MEDLINE OVID, DIALNET and the COCHRANE LIBRARY. After analyzing the texts, eleven studies aimed directly at verifying the influence of grandmothers on the incidence and prevalence of breastfeeding in different contexts were selected. Results: The grandmother emerges as a source of support among the rural population (41%). In the urban population, the grandmother appears among the three main factors (mother's intention to breastfeed or not, mother's decision on a feeding pattern for her child, and objective information on breastfeeding patterns), in a supprting role (94, 7%) and in a hindering role (26.7%). As for the mother's decision on a feeding pattern for her children, 78% reported having made the decision before pregnancy or in the first quarter. Living with the grandmother is positively associated with providing water or teas (PR in Cox regression with 95% CI: 1.51 [1.10 to 2.09], p = 0.011). Conclusions: In the light of the review, the detracting or non-detracting influence exerted by the grandmother with respect to breastfeeding affects the three main factors. The detracting influence exerted by the grandmother's advice seems to be related to incorrect knowledge acquired more as part of the "bottle fed generation" rather than intentionality, and is susceptible to being modified through appropriate information, with favorable results on the feeding pattern.

Objetivo: pesquisar a influência da avó sobre a mãe que dá lactancia materna e sua relação com a incidência e prevalência desta. Materiais e métodos: realizou-se uma revisão integral consultando as bases de dados MEDLINE OVID, DIALNET e COCHRANE LIBRARY. Após a análise dos textos, escolheram-se onze estudos de pesquisa que vão dirigidos diretamente a comprovar a influência das avós na incidência e prevalência da lactancia materna em diferentes contextos. Resultados: constatou-se que a avó apareceu como fonte de apoio na população rural (41%). Na população urbana, a avó aparece entre os três fatores principais (intenção da mãe de lactar ou não, decisão da mãe do padrão de alimentação de seu filho e informação objetiva sobre pautas de amamentação), tanto no papel de apoio (94,7%) quanto no de obstáculo (26,7%). Quanto à decisão da mãe sobre o padrão de alimentação de seus filhos, 78% referem ter tomado a decisão antes da gravidez ou no primeiro trimestre. A convivência com a avó teve associação positiva com dar água ou infusões (PR regressão de Cox com IC 95%: 1,51 [1,10-2,09]; p = 0,011). Conclusões: à luz da revisão, a influência detratora ou não que exerce a avó sobre a lactancia materna se dá sobre os três fatores principais. A influência detratora exercida pelos conselhos avó parece estar mais relacionada com conhecimentos errôneos adquiridos como "geração da mamadeira" do que com a intencionalidade e é suscetível de ser modificado com informação adequada, com resultados favoráveis sobre o padrão de alimentação.

Long-chain fatty alcohols from pomace olive oil modulate the release of proinflammatory mediators.

Pomace olive oil is a by-product of olive oil extraction that is traditionally produced and consumed in Spain. The nonglyceride matter of this oil is a good source of interesting minor compounds, like long-chain fatty alcohols, which are present free or as part of waxes. In the present study, long-chain fatty alcohols were isolated from the nonglyceride fraction of pomace olive oil, and the composition was identified and quantified. The major components of long-chain fatty alcohols were tetracosanol, hexacosanol and octacosanol. We investigated the ability of long-chain fatty alcohols from pomace olive oil to inhibit the release of different proinflammatory mediators in vitro by cells involved in inflammatory processes. Long-chain fatty alcohols significantly and dose-dependently decreased nitric oxide production by RAW 264.7 murine macrophages stimulated with lipopolysaccharide. Western blot analysis showed that nitric oxide reduction was a consequence of the inhibition of inducible nitric oxide synthetase expression. Long-chain fatty alcohols also reduced tumor necrosis factor-alpha and prostaglandin E(2) production, although the potency of inhibition for the latter was lower. On the other hand, long-chain fatty alcohols significantly reduced thromboxane A(2) production in rat peritoneal neutrophils stimulated with the calcium ionophore A-23187. The reduction of eicosanoid release was related to the inhibition of phospholipase A(2) enzyme activity by long-chain fatty alcohols, reaching an inhibitory concentration 50% value of 6.2 microg/ml. These results showed that long-chain fatty alcohols may have a protective effect on some mediators involved in the inflammatory damage development, suggesting its potential value as a putative functional component of pomace olive oil.

Supressive effect of maslinic acid from pomace olive oil on oxidative stress and cytokine production in stimulated murine macrophages.

The pentacyclic triterpene maslinic acid (MA) is a natural compound present in the non glyceride fraction of pomace olive oil, also called orujo olive oil. This compound has previously demonstrated antioxidant properties against lipid peroxidation in vitro, but its effects on reactive oxygen and nitrogen-derived species and pro-inflammatory cytokines generated by a cell system have not yet been investigated. In this study, we have tested the effect of MA upon oxidative stress and cytokine production using peritoneal murine macrophages. MA significantly inhibited the enhanced production of nitric oxide (NO) induced by lypopolysaccharide (LPS) when it was measured by the nitrite production with an inhibitory concentration 50% value (IC(50)) of 25.4 microM. This inhibiting effect seems to be consequence of an action at the level of the LPS-induction of the inducible nitric oxide synthethase (iNOS) gene enzyme expression rather than to a direct inhibitory action on enzyme activity. The secretion of the inflammatory cytokines interleukine-6 and TNF-a from LPS-stimulated murine macrophages was also significantly reduced (p < 0.05 and 0.01) by 50 and 100 microM of MA. In addition, reactive oxygen species were measured after stimulation with phorbol-12-myristate-13-acetate (PMA). Thus, pre-treatment with MA reduced the generation of hydrogen peroxide from stimulated macrophages in a dose-dependent manner (IC(50): 43.6 microM) as assayed by the oxidation of the peroxidase enzyme. However, no inhibitory effect on superoxide release, measured by the reduction of ferricytochrome c, was observed after the pretreatment with MA in the culture medium. These results suggest a potential biopharmaceutical use of this hydroxy-pentacyclic triterpene derivative, present in orujo olive oil, on preventing oxidative stress and pro-inflammatory cytokine generation.

Aspectos psicológicos del invidente / Psychological features in blindness

La ceguera es uno de lo problemas de salud más incapacitantes en el ser humano, siendo extraordinariamente difícil el obtener estadística respecto al número, características demográficas y causas de ceguera, y aún más el entender los componentes psicológicos que resultan de esta condición. Se estima que el número de invidentes en el mundo es de alrededor de 27 a 35 millones, de los cuales 500 mil corresponden a población mexicana. Existen características psicológicas que afectan no sólo al invidente, repercutiendo también en áreas familiar y social y dada la creciente incidencia del problema por aumento de la edad poblacional y espectativa de vida, se hace aún más necesario abordarlo en forma multicisciplinaria. el objetivo del presente trabajo es difundir el conocimiento actual respecto a los aspectos psicológicos del invidente y sentar bases para futuras investigaciones