Measurement of the specific and non-specific binding energies of Mg2+ to RNA.

Determining the non-specific and specific electrostatic contributions of magnesium binding to RNA is a challenging problem. We introduce a single-molecule method based on measuring the folding energy of a native RNA in magnesium and at its equivalent sodium concentration. The latter is defined so that the folding energy in sodium equals the non-specific electrostatic contribution in magnesium. The sodium equivalent can be estimated according to the empirical 100/1 rule (1 M NaCl is equivalent to 10 mM MgCl2), which is a good approximation for most RNAs. The method is applied to an RNA three-way junction (3WJ) that contains specific Mg2+ binding sites and misfolds into a double hairpin structure without binding sites. We mechanically pull the RNA with optical tweezers and use fluctuation theorems to determine the folding energies of the native and misfolded structures in magnesium (10 mM MgCl2) and at the equivalent sodium condition (1 M NaCl). While the free energies of the misfolded structure are equal in magnesium and sodium, they are not for the native structure, the difference being due to the specific binding energy of magnesium to the 3WJ, which equals ΔG≃ 10 kcal/mol. Besides stabilizing the 3WJ, Mg2+ also kinetically rescues it from the misfolded structure over timescales of tens of seconds in a force-dependent manner. The method should generally be applicable to determine the specific binding energies of divalent cations to other tertiary RNAs.

Are high-sensitivity troponins always reliable? Donor-recipient troponin transfusion in liver transplantation.

Blood troponins are used to diagnose perioperative myocardial injury and infarction. During liver transplantation, a passive donor-recipient troponin transfer with the graft may result in an increase of troponins in the transplant recipient questioning the diagnosis of myocardial injury. We present a case of liver transplantation with sudden elevation of recipient's serum troponin levels immediately after graft reperfusion and its subsequent normalization in which myocardial damage and other non-ischaemic potential causes were ruled out. Patient consent for publication was obtained prior to submission of the manuscript.

Circulating leptin and adiponectin, and breast density in premenopausal Mexican women: the Mexican Teachers' Cohort.

PURPOSE: Leptin and adiponectin are produced by the adipose tissue. Mammographic density (MD) is one of the strongest predictors of breast cancer (BC) and is highly influenced by adiposity. How the interplay between MD, obesity, and obesity-related biomarkers influences BC risk, however, is still unknown, especially in premenopausal women, where adiposity seems to be protective for BC. The aim of the present study was to explore the association between circulating leptin, adiponectin, and their ratio, with MD in Mexican premenopausal women who are part of the large Mexican Teachers' Cohort (MTC). METHODS: A subsample of 2,084 women from the MTC participated in a clinical evaluation. Of them, 574 premenopausal women were randomly selected, from four MD strata. Serum leptin and adiponectin concentrations were measured by immunoassays. Multivariate regression analyses were performed to compare means of MD by quartiles of adipokines and their ratio. RESULTS: High leptin and leptin/adiponectin ratio levels were significantly associated with lower percentage MD and higher absolute and non-absolute dense tissue areas. High adiponectin levels were significantly associated with lower absolute dense and non-dense tissue areas, but not with percentage MD. After adjustment for BMI, only the associations between percentage MD and absolute non-dense tissue area with leptin remained statistically significant. CONCLUSIONS: Leptin, adiponectin, and their ratio were associated with MD; however, only the positive association with leptin seemed to be independent from overall obesity.

[Impact of COVID-19 lockdown on patients with idiopathic intracranial hypertension in Spain]. / Impacto del confinamiento por COVID-19 en pacientes con hipertensión intracraneal idiopática en España.

INTRODUCTION: The nationwide lockdown implemented in Spain in March 2020 in response to the 2019 coronavirus disease pandemic (COVID-19) could have had an effect on the clinical situation, disease management and access to care in patients with idiopathic intracranial hypertension. PATIENTS AND METHODS: A cross-sectional observational study was conducted by means of an online survey. The frequency and impact of headache, visual symptoms, dizziness, cognitive symptoms, diplopia, anxiety and depression on patients' quality of life were recorded. Disease treatment and delays experienced in accessing care were recorded. Participants were grouped according to the time elapsed since diagnosis. RESULTS: A total of 112 patients participated in the study (103 women). The average time since disease onset was 6.25 years. Headache, anxiety and depression were responsible for considerable impact in the period prior to lockdown. The impact of headache was not aggravated during lockdown, but anxiety and depression did become significantly worse. Recently diagnosed participants reported higher rates of anxiety, depression, dizziness and diplopia; chronically diagnosed participants reported a higher frequency of cognitive symptoms. CONCLUSIONS: The clinical situation of the participants deteriorated during lockdown, although the differences were smaller than previously thought. This was probably because the baseline situation was more severe than expected. Further studies are needed to clarify the medium and long-term impact of the COVID-19 pandemic on patients with idiopathic intracranial hypertension.

TITLE: Impacto del confinamiento por COVID-19 en pacientes con hipertensión intracraneal idiopática en España.Introducción. El confinamiento nacional instaurado en España en marzo de 2020 como respuesta a la pandemia por la enfermedad por coronavirus 2019 (COVID-19) pudo tener un efecto en la situación clínica, el manejo de la enfermedad y el acceso a la atención médica en pacientes con hipertensión intracraneal idiopática. Pacientes y métodos. Se realizó un estudio observacional transversal por medio de una encuesta en línea. Se registraron la frecuencia y el impacto en la calidad de vida de la cefalea, los síntomas visuales, el mareo, los síntomas cognitivos, la diplopía, la ansiedad y la depresión. Se registró el tratamiento de la enfermedad y los retrasos experimentados en el acceso a la atención médica. Se agrupó a los participantes según el tiempo de evolución desde el diagnóstico. Resultados. Participaron 112 pacientes (103 mujeres). El tiempo medio de evolución de la enfermedad fue 6,25 años. La cefalea, la ansiedad y la depresión fueron responsables de un impacto considerable en el período previo al confinamiento. El impacto de la cefalea no empeoró durante el confinamiento, pero la ansiedad y la depresión empeoraron significativamente. Los participantes diagnosticados recientemente comunicaron mayores tasas de ansiedad, depresión, mareo y diplopía; los de evolución crónica, mayor frecuencia de síntomas cognitivos. Conclusiones. La situación clínica de los participantes se deterioró durante el confinamiento, aunque las diferencias fueron menores de lo que se pensaba. Esto se debió, probablemente, a que la situación basal era más grave de lo esperado. Son necesarios más estudios para aclarar el impacto a medio y largo plazo de la pandemia por COVID-19 en pacientes con hipertensión intracraneal idiopática.

Reliability and validity of the Mexican teachers' physical activity questionnaire (MTPAQ) in a subsample of female Mexican teachers.

BACKGROUND: Reliable and valid instruments are needed to estimate physical activity levels. The purposes of this study were to estimate the reliability and validity of the Physical Activity Questionnaire (MTPAQ) in a subsample of the Mexican Teachers Cohort study. METHODS: We completed telephone interviews and clinical examinations of 82 teachers. Two MTPAQ, five International Physical Activity Questionnaire (IPAQ)-long form, and two accelerometer (AC) measures were used to determine physical activity levels throughout 24 months. Moderate and walking physical activity (MWPA min/week), vigorous physical activity (VPA min/week), and moderate-to-vigorous physical activity minutes per week (MVPA min/week) were estimated for each instrument. Pearson, Intra-class correlations and deattenuated adjustments were used to determine the reliability and validity of MTPAQ. RESULTS: MWPA and MVPA min/week of MTPAQs were moderately correlated (r ≥ 0.54) to min/week of IPAQ-long form. MWPA and MVPA min/week average MTPAQ and MTPAQ1 and average AC, AC1 and AC2 were fairly correlated (r ≥ 0.20). A higher correlation was observed when correlation coefficients were attenuated (r ≥ 0.32). CONCLUSIONS: MTPAQ1 is a reliable and valid tool to measure physical activity levels.

Dynamics of cluster-forming hub-filament systems: The case of the high-mass star-forming complex Monoceros R2.

CONTEXT: High-mass stars and star clusters commonly form within hub-filament systems. Monoceros R2 (hereafter Mon R2), at a distance of 830 pc, harbors one of the closest such systems, making it an excellent target for case studies. AIMS: We investigate the morphology, stability and dynamical properties of the Mon R2 hub-filament system. METHODS: We employ observations of the 13CO and C18O 1→0 and 2→1 lines obtained with the IRAM-30m telescope. We also use H2 column density maps derived from Herschel dust emission observations. RESULTS: We identified the filamentary network in Mon R2 with the DisPerSE algorithm and characterized the individual filaments as either main (converging into the hub) or secondary (converging to a main filament) filaments. The main filaments have line masses of 30-100 M ⊙ pc-1 and show signs of fragmentation, while the secondary filaments have line masses of 12-60 M ⊙ pc-1 and show fragmentation only sporadically. In the context of Ostriker's hydrostatic filament model, the main filaments are thermally supercritical. If non-thermal motions are included, most of them are trans-critical. Most of the secondary filaments are roughly transcritical regardless of whether non-thermal motions are included or not. From the morphology and kinematics of the main filaments, we estimate a mass accretion rate of 10-4-10-3 M ⊙ yr-1 into the central hub. The secondary filaments accrete into the main filaments with a rate of 0.1-0.4×10-4 M ⊙ yr-1. The main filaments extend into the central hub. Their velocity gradients increase towards the hub, suggesting acceleration of the gas.We estimate that with the observed infall velocity, the mass-doubling time of the hub is ~ 2:5 Myr, ten times larger than the free-fall time, suggesting a dynamically old region. These timescales are comparable with the chemical age of the Hii region. Inside the hub, the main filaments show a ring- or a spiral-like morphology that exhibits rotation and infall motions. One possible explanation for the morphology is that gas is falling into the central cluster following a spiral-like pattern.

Impacto del confinamiento por COVID-19 en pacientes con hipertensión intracraneal idiopática en España / Impact of COVID-19 lockdown on patients with idiopathic intracranial hypertension in Spain

Introducción: El confinamiento nacional instaurado en España en marzo de 2020 como respuesta a la pandemia por la enfermedad por coronavirus 2019 (COVID-19) pudo tener un efecto en la situación clínica, el manejo de la enfermedad y el acceso a la atención médica en pacientes con hipertensión intracraneal idiopática. Pacientes y métodos: Se realizó un estudio observacional transversal por medio de una encuesta en línea. Se registraron la frecuencia y el impacto en la calidad de vida de la cefalea, los síntomas visuales, el mareo, los síntomas cognitivos, la diplopía, la ansiedad y la depresión. Se registró el tratamiento de la enfermedad y los retrasos experimentados en el acceso a la atención médica. Se agrupó a los participantes según el tiempo de evolución desde el diagnóstico. Resultados: Participaron 112 pacientes (103 mujeres). El tiempo medio de evolución de la enfermedad fue 6,25 años. La cefalea, la ansiedad y la depresión fueron responsables de un impacto considerable en el período previo al confinamiento. El impacto de la cefalea no empeoró durante el confinamiento, pero la ansiedad y la depresión empeoraron significativamente. Los participantes diagnosticados recientemente comunicaron mayores tasas de ansiedad, depresión, mareo y diplopía; los de evolución crónica, mayor frecuencia de síntomas cognitivos. Conclusiones: La situación clínica de los participantes se deterioró durante el confinamiento, aunque las diferencias fueron menores de lo que se pensaba. Esto se debió, probablemente, a que la situación basal era más grave de lo esperado. Son necesarios más estudios para aclarar el impacto a medio y largo plazo de la pandemia por COVID-19 en pacientes con hipertensión intracraneal idiopática.(AU)

Introduction: The nationwide lockdown implemented in Spain in March 2020 in response to the 2019 coronavirus disease pandemic (COVID-19) could have had an effect on the clinical situation, disease management and access to care in patients with idiopathic intracranial hypertension. Patients and methods: A cross-sectional observational study was conducted by means of an online survey. The frequency and impact of headache, visual symptoms, dizziness, cognitive symptoms, diplopia, anxiety and depression on patients’ quality of life were recorded. Disease treatment and delays experienced in accessing care were recorded. Participants were grouped according to the time elapsed since diagnosis. Results: A total of 112 patients participated in the study (103 women). The average time since disease onset was 6.25 years. Headache, anxiety and depression were responsible for considerable impact in the period prior to lockdown. The impact of headache was not aggravated during lockdown, but anxiety and depression did become significantly worse. Recently diagnosed participants reported higher rates of anxiety, depression, dizziness and diplopia; chronically diagnosed participants reported a higher frequency of cognitive symptoms. Conclusions: The clinical situation of the participants deteriorated during lockdown, although the differences were smaller than previously thought. This was probably because the baseline situation was more severe than expected. Further studies are needed to clarify the medium and long-term impact of the COVID-19 pandemic on patients with idiopathic intracranial hypertension.(AU)

[Transplantation of neural stem cells into the cochlea]. / Neurogene Stammzelltransplantation in die Kochlea.

BACKGROUND: Stem cell therapy is especially interesting for inner ear related diseases, since the hair cells are very sensitive and do not regenerate. Hair cell loss is therefore irreversible and is accompanied by hearing loss. In the last few years, different research groups have transplanted stem cells into the inner ear with promising results. In the presented study, our aim was to gain insight into how neuronal stem cells behave when they are transplanted, both in vitro and in vivo, into a damaged inner ear. METHODS: Neuronal stem cells from E9.5 day old mouse embryos were collected and infected with an adenoviral vector encoding green fluorescent protein (GFP). GFP+ cells were then transplanted into a damaged organ of Corti in vitro or into a damaged mouse inner ear in vivo. RESULTS: We were able to detect GFP+ cells close to the organ of Corti in vitro and in the organ of Corti in vivo. The GFP+ cells do not seem to be randomly distributed in either the in vitro or in vivo situation. Most interestingly, GFP+ cells could be detected close to places where hair cells had been lost in vivo. CONCLUSION: Neuronal stem cells are interesting candidates to replace lost hair cells. However, a great deal of research is still needed before they can enter clinical trials.

Aprendizaje expandido: gestión de una comunidad de práctica online para educadores físicos / Expanded Learning Process in Managing a PE Teachers’ On-Line Community of Practice

A través de un estudio de caso, se indaga en las tensiones detectadas en el diseño y gestión de una comunidad de práctica online de profesorado de Educación Física. El marco conceptual usado en la discusión es el “Aprendizaje Expandido” (Engreström). Se constata un modelo de cinco fases. En la fase 1, las tensiones se dan entre las ideas de los propios diseñadores; en la fase 2, entre las ideas y las posibilidades tecnológicas; en la fase 3, las tensiones surgen entre las ideas de los desarrolladores, las prácticas y las necesidades de los participantes; en la fase 4, continúa la tensión entre desarrolladores y prácticas, mediatizadas por la incorporación de nuevos gestores que provienen de la comunidad de práctica; y, finalmente, en la fase 5, los diseñadores y gestores van retirándose del primer plano de la plataforma y su ideas iniciales acaban disolviéndose entre las de la comunidad. (AU)

Through a case study, tensions detected in the design and management of an online community of practice for Physical Education teachers are investigated. The discussion is established within the conceptual framework of "Expanded Learning" (Engreström). A five-phase model was found. In phase 1, tensions arise between the ideas of the designers themselves; in phase 2, tensions are established between ideas and technological possibilities; in phase 3, tensions arise between developers' ideas, practices, and participants’ needs; in phase 4, tensions between developers and practices continue, mediated by the incorporation of new managers who come from the community of practice; and finally, in phase 5, designers and managers quit the forefront of the platform and their initial ideas end up dissolving among the ideas of the community. (AU)

(sub)Millimeter Emission Lines of Molecules in Born-again Stars⋆.

CONTEXT: Born-again stars offer us a unique possibility of studying the evolution of the circumstellar envelope of evolved stars in human timescales. To present, most of the observations of the circumstellar material in these stars have been limited to study the relatively hot gas and dust. In other evolved stars, the emission from rotational transitions of molecules, such as CO, is commonly used to study the cool component of their circumstellar envelopes. Thus, the detection and study of molecular gas in born-again stars would be of great importance to understand their composition and chemical evolution. In addition, the molecular emission would be an invaluable tool to explore the physical conditions, kinematics and formation of asymmetric structures in the circumstellar envelopes of these evolved stars. However, until now, all attempts to detect molecular emission from the cool material around born-again stars have failed. AIMS: We searched for emission from rotational transitions of molecules in the hydrogen-deficient circumstellar envelopes of born-again stars to explore the chemical composition, kinematics, and physical parameters of the relatively cool gas. METHODS: We carried out observations using the APEX and IRAM 30m telescopes to search for molecular emission toward four well studied born-again stars, V4334 Sgr, V605 Aql, A30 and A78, that are thought to represent an evolutionary sequence. RESULTS: We detected for the first time emission from HCN and H13CN molecules toward V4334 Sgr, and CO emission in V605 Aql. No molecular emission was detected above the noise level toward A30 and A78. The detected lines exhibit broad linewidths ≳150 km s-1, which indicates that the emission comes from gas ejected during the born-again event, rather than from the old planetary nebula. A first estimate of the H12CN/H13CN abundance ratio in the circumstellar environment of V4334 Sgr is ≈3, which is similar to the value of the 12C/13C ratio measured from other observations. We derived a rotational temperature of T rot=13±1 K, and a total column density of N HCN=1.6±0.1×1016 cm-2 for V4334 Sgr. This result sets a lower limit on the amount of hydrogen that was ejected into the wind during the born-again event of this source. For V605 Aql, we obtained a lower limit for the integrated line intensities I 12C/I 13C>4.

Far-infrared observations of a massive cluster forming in the Monoceros R2 filament hub⋆.

We present far-infrared observations of Monoceros R2 (a giant molecular cloud at approximately 830 pc distance, containing several sites of active star formation), as observed at 70 µm, 160 µm, 250 µm, 350 µm, and 500 µm by the Photodetector Array Camera and Spectrometer (PACS) and Spectral and Photometric Imaging Receiver (SPIRE) instruments on the Herschel Space Observatory as part of the Herschel imaging survey of OB young stellar objects (HOBYS) Key programme. The Herschel data are complemented by SCUBA-2 data in the submillimetre range, and WISE and Spitzer data in the mid-infrared. In addition, C18O data from the IRAM 30-m Telescope are presented, and used for kinematic information. Sources were extracted from the maps with getsources, and from the fluxes measured, spectral energy distributions were constructed, allowing measurements of source mass and dust temperature. Of 177 Herschel sources robustly detected in the region (a detection with high signal-to-noise and low axis ratio at multiple wavelengths), including protostars and starless cores, 29 are found in a filamentary hub at the centre of the region (a little over 1% of the observed area). These objects are on average smaller, more massive, and more luminous than those in the surrounding regions (which together suggest that they are at a later stage of evolution), a result that cannot be explained entirely by selection effects. These results suggest a picture in which the hub may have begun star formation at a point significantly earlier than the outer regions, possibly forming as a result of feedback from earlier star formation. Furthermore, the hub may be sustaining its star formation by accreting material from the surrounding filaments.

Design and evaluation of "3 + 1" mixed ligand oxorhenium and oxotechnetium complexes bearing a nitroaromatic group with potential application in nuclear medicine oncology.

The synthesis and evaluation of a series of oxotechnetium and oxorhenium complexes containing a nitroaromatic moiety as potential radiopharmaceuticals for targeting tumour hypoxia is presented. 99mTc labelling was performed in high yield (>85%) and radiochemical purity (>90%). Their structure was corroborated by means of the rhenium complexes. Reduction potentials were in the range for bioreducible compounds. 99mTc complexes III-VI were selected for "in vivo" experiments in view of the results of cytotoxicity studies. Biodistribution in normal animals was characterized by high initial blood, lung and liver uptake, fast blood and soft tissue depuration and preferential excretion via the hepatobiliary system. Initial tumour uptake was moderate but tumour/muscle ratios for complexes III and IV, were favourable at all time points. Although the results are encouraging further development is still necessary in order to achieve higher tumour uptake and lower gastrointestinal activity.

Bovine herpesvirus 4-associated postpartum metritis in a Spanish dairy herd.

In more than 10 Spanish dairy cows, a bovine herpesvirus 4 (BHV4) associated postpartum metritis was confirmed by virus isolation, BHV4-glycoprotein B (gB) PCR and/or serology. In this study, 12 cows with, and, at the time of sampling, 3 cows without clinical signs of acute postpartum metritis from one large dairy herd in Spain were examined for bacterial and viral infections. Blood, placenta/caruncles and uterine contents were collected between day 1 and day 20 post-calving, and examined for the presence of bacteria and for viruses by virus isolation, BHV4 DNA by BHV4-gB PCR and/or BHV4 antibody titres. Bovine herpesvirus 4 was detected in 83% of the cases with clinical signs of acute postpartum metritis by virus isolation and/or BHV4-gB PCR. An increase of BHV4 antibodies was detected in all examined postpartum metritis cows and in the 3 cows without clinical metritis. Two of these 3 cows developed severe metritis a few dayss after collecting the first blood sample. A concurrent infections of BHV4 and bacteria, mainly Arcanobacterium pyogenes and Streptococcus sp., were detected in 73% of the examined uterine contents collected from postpartum metritis affected cows. This case-report study showed a clear association between BHV4 infections and acute postpartum metritis in dairy cows. In addition, the BHV4-associated postpartum metritis appeared to be an emerging syndrome in this Spanish herd.

Phenazine 5,10-dioxide derivatives as hypoxic selective cytotoxins: Part II. Structure-activity relationship studies.

The synthesis and evaluation as hypoxic selective cytotoxins of new derivatives of 2-amino or 2-hydroxyphenazine 5,10-dioxide are described. The compounds were developed as structural analogs of other bioreductive compounds and its in vitro cytotoxicities on V79 cells under hypoxic and aerobic conditions were determined. To gain insight into its mechanism of action electrochemical behavior, interaction with DNA experiments and QSAR studies were performed.

The first CO+ image: I. Probing the HI/H2 layer around the ultracompact HII region Mon R2.

The CO+ reactive ion is thought to be a tracer of the boundary between a HII region and the hot molecular gas. In this study, we present the spatial distribution of the CO+ rotational emission toward the Mon R2 star-forming region. The CO+ emission presents a clumpy ring-like morphology, arising from a narrow dense layer around the HII region. We compare the CO+ distribution with other species present in photon-dominated regions (PDR), such as [CII] 158 µm, H2 S(3) rotational line at 9.3 µm, polycyclic aromatic hydrocarbons (PAHs) and HCO+. We find that the CO+ emission is spatially coincident with the PAHs and [CII] emission. This confirms that the CO+ emission arises from a narrow dense layer of the HI/H2 interface. We have determined the CO+ fractional abundance, relative to C+ toward three positions. The abundances range from 0.1 to 1.9 ×10-10 and are in good agreement with previous chemical model, which predicts that the production of CO+ in PDRs only occurs in dense regions with high UV fields. The CO+ linewidth is larger than those found in molecular gas tracers, and their central velocity are blue-shifted with respect to the molecular gas velocity. We interpret this as a hint that the CO+ is probing photo-evaporating clump surfaces.

Computer modeling of protein folding: conformational and energetic analysis of reduced and detailed protein models.

Recently we developed methods to generate low-resolution protein tertiary structures using a reduced model of the protein where secondary structure is specified and a simple potential based on a statistical analysis of the Protein Data Bank is employed. Here we present the results of an extensive analysis of a large number of detailed, all-atom structures generated from these reduced model structures. Following side-chain addition, minimization and simulated annealing simulations are carried out with a molecular mechanics potential including an approximate continuum solvent treatment. By combining reduced model simulations with molecular modeling calculations we generate energetically competitive, plausible misfolded structures which provide a more significant test of the potential function than current misfolded models based on superimposing the native sequence on the folded structures of completely different proteins. The various contributions to the total energy and their interdependence are analyzed in detail for many conformations of three proteins (myoglobin, the C-terminal fragment of the L7/L12 ribosomal protein, and the N-terminal domain of phage 434 repressor). Our analysis indicates that the all-atom potential performs reasonably well in distinguishing the native structure. It also reveals inadequacies in the reduced model potential, which suggests how this potential can be improved to yield greater accuracy. Preliminary results with an improved potential are presented.

First record of Porocephalus cf. clavatus (Pentastomida: Porocephalida) as a parasite on Bothrops asper (Squamata: Viperidae) in Costa Rica.

Pentastomids are parasites that infect respiratory cavities of vertebrates, they are pretty common but poorly known in wildlife veterinary. A Bothrops asper snake (Garman, 1884) was captured in the Caribbean region of Costa Rica and had its lung infested with pentastomids, identified as ca Porocephalus clavatus (Wyman, 1845). This represents the first record of Porocephalus (Humboldt, 1812) on B. asper as well as P. cf. clavatus in Costa Rica. Further studies are needed to clarify their taxonomic position, images and scanning electron microscopy photographs (SEM) of the specimens are given.

Pharmacodynamics of subcutaneous recombinant human interleukin-10 in healthy volunteers.

Interleukin-10 inhibits T-lymphocyte activation and proliferation and lipopolysaccharide-induced monocyte production of proinflammatory cytokines. Fifty-four healthy volunteers received single doses of recombinant human interleukin-10 (1.0, 2.5, 5.0, 10, 25, or 50 micrograms/kg) or placebo by subcutaneous injection (randomized double-blind assignment). Clinical adverse events were infrequent at doses below 50 micrograms/kg (five of six subjects had mild flu-like syndrome). Mean serum interleukin-10 concentrations were dose related. The mean terminal-phase half-life ranged from 2.7 to 4.5 hours, and the apparent volume of distribution ranged from 0.70 to 1.35 L/kg. Hematologic changes included transient mild to moderate increases of neutrophil counts, decreases of lymphocyte counts, and a delayed decrease of platelet counts. Recombinant human interleukin-10 significantly suppressed production of the proinflammatory cytokines interleukin-1 beta and tumor necrosis factor-alpha by whole blood stimulated ex vivo with Escherichia coli lipopolysaccharide.

Selective thromboxane synthetase inhibitors and antihypertensive agents. New derivatives of 4-hydrazino-5H-pyridazino[4,5-b]indole, 4-hydrazinopyridazino[4,5-a]indole, and related compounds.

A series of new derivatives of 4-hydrazino-5H-pyridazino[4,5-b]indole (5) and 4-hydrazinopyridazino[4,5-a]indole (12) have been synthesized to investigate their activities as selective thromboxane synthetase inhibitors as well as antihypertensive agents. Several of the prepared compounds were found to be selective thromboxane synthetase inhibitors, in concordance with the Gorman model. The most potent were 8-(benzyloxy)-3,4-dihydro-4-oxo-5H-pyridazino[4,5-b]indole (3c) and 8-methoxy-4-hydrazino-5H-pyridazino[4,5-b]indole (5). This last compound did not inhibit prostacyclin formation and showed an antihypertensive activity similar to that of hydralazine. The acute toxicity in mice for 5a . HCl is about 2.2 times less than that for hydralazine.

Hypoxia-selective agents derived from 2-quinoxalinecarbonitrile 1,4-di-N-oxides. 2.

Hypoxic cells are an important target for antitumor therapy because tumors are typically characterized by such cells. Virtually all tumors which are present as solid masses contain hypoxic cells, while normal cells generally have an adequate supply of oxygen. Accordingly, antitumor agents can be made selective for tumors by virtue of high activity under hypoxic conditions. The initial purpose of this work was to determine the influence of different groups in position 3. Thus, the synthesis of some 3-NH-substituted derivatives (2a, 3a, 4a) starting from 3-amino-2-quinoxalinecarbonitrile 1,4-di-N-oxide (1a) is described. Reductive deamination of compounds 1a-k provides the 2-quinoxalinecarbonitriles 5a-k, which are more potent, while selectivity is maintained or increased in some derivatives. The compound 7-(4-nitrophenyl)-2-quinoxalinecarbonitrile 1,4-di-N-oxide (5k) is 150-fold more potent than tirapazamine (3-amino-1,2,4- benzotriazine 1,4-di-N-oxide), which has been used as a standard. Three derivatives (5g,i,k) show a hypoxic cytotoxicity ratio (HCR) > or = 200, better than that of tirapazamine (HCR = 75) in V79 cells. Replacement of the 3-amino group by chlorine affords the potent but nonselective 3-chloro derivatives 6a-k showing similar toxicities under both aerobic and hypoxic conditions. These compounds were used as intermediates for the synthesis of a new series of water-soluble compounds derived from 3-[[(N,N- dialkylamino)alkyl[amino]-2-quinoxalinecarbonitrile 1,4-di-N-oxides 10a-i and 11a-i. The 7-chloro and the 7-trifluoromethyl derivatives 10b,f have demonstrated high potency (0.4 and 0.3 microM) and excellent selectivity (HCR = 250 and 340). Several 7-chloro analogues, 12b, 13b.1,b.2, and 14b, and the dimer 16b have been prepared and evaluated in order to determine the optimum lateral chain in position 3, which appears to be the [(N,N-dimethylamino)propyl]amino moiety.