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The application of geoconservation concepts and methodologies to the Azores archipelago led to the implementation of the Geopark Azores, recognized as such by the European and Global Geoparks Network. The current work re-evaluates and stresses the scientific and touristic value of the palaeontological sites of Santa Maria Island. Two new geosites (the Ponta do Castelo tempestite deposit and the Pedra-que-pica coquina) are proposed for classification as ‘Regional Natural Monuments’ by the Regional Government of the Azores, due to their international relevance. The tempestite deposit of Ponta do Castelo was overlain by a contemporary coastal lava delta, which enables the inference of the precise water depth of the geosite at the time of deposition, a very rare condition worldwide; and Pedra-que-pica is the most extensive multispecific fossiliferous coquina ever reported in the literature from the shelf of any of the ∼20,000 known volcanic oceanic islands in the world. Relevant geosites reported for this island are increased from 15 to 26. Additional palaeontological heritage contributions to the sustainable tourism of Santa Maria are suggested, with a focus on two recent projects: the ‘Fossil Trail’ and the future ‘PalaeoPark Santa Maria’.

In the field of traffic management and control systems, we are witnessing a symbiotic evolution, where intelligent infrastructure is progressively collaborating with smart vehicles to produce benefits for traffic monitoring and security, by rapidly identifying hazardous behaviours. This exponential growth is due to the rapid development of deep learning in recent years, as well as the improvements in computer vision models. These technologies allow for monitoring tasks without the need to install numerous sensors or stop the traffic, using the extensive camera network of surveillance cameras already present in worldwide roads. This study proposes a computer vision-based solution that allows for real-time processing of video streams through edge computing devices, eliminating the need for Internet connectivity or dedicated sensors. The proposed system employs deep learning algorithms and vision techniques that perform vehicle detection, classification, tracking, speed estimation, and vehicle geolocation.

The two-spotted spider mite (TSSM) Tetranychus urticae is a polyphagous pest which infests several wild and cultivated species of plants worldwide. To date, this mite is the most deleterious pest attacking the strawberry plant under a protected environment. Exogenously applied salicylic acid (SA) has been found to induce resistance to biotic and abiotic stresses. Morpho-anatomical changes in strawberry leaflets and TSSM preference tests were investigated in response to SA treatment. Plants grown in a greenhouse were foliar sprayed with different concentrations of SA (0 mg · L−1, 25 mg · L−1, 50 mg · L−1, 75 mg · L−1 and 100 mg · L−1). After the third application, certain parameters including the number of glandular and non-glandular trichomes, thickness of leaflet, abaxial cell wall, adaxial cell wall, palisade and lacunous parenchyma and mesophyll were measured using scanning electronic microscopy (SEM). Two-choice and non-choice assays were employed to verify the TSSM preference. Exogenous treatment with SA promoted morpho-anatomical changes in the following parameters, namely: thickness of the leaflets, mesophyll, lacunous and palisade parenchyma, cell wall (abaxial and adaxial) and the number of glandular and non-glandular trichomes in strawberry leaflets. In general, TSSM preferred less leaflets treated with SA compared with the control in a two-choice assay. A lesser number of TSSM eggs and live females were also recorded in leaflets treated with SA compared with the control plants in no-choice assays. The values of the number of eggs and live females correlated negatively with those obtained for the morpho-anatomical traits induced by exogenous SA.

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Copyright of Estudios Demográficos y Urbanos is the property of El Colegio de Mexico AC and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Business Process Modelling (BPM) is one of the most important phases of information system design. Business Process (BP) meta-models allow capturing informational and behavioural aspects of business processes. Unfortunately, standard BP meta-modelling approaches focus just on process description, providing different BP models. It is not possible to compare and identify related daily practices in order to improve BP models. This lack of information implies that further research in BP meta-models is needed to reflect the evolution/change in BP. Considering this limitation, this paper introduces a new BP meta-model designed by Business Process and Practice Alignment Meta-model (BPPAMeta-model). Our intention is to present a meta-model that addresses features related to the alignment between daily work practices and BP descriptions. Objectives: This paper intends to present a metamodel which is going to integrate daily work information into coherent and sound process definitions. Methods/Approach: The methodology employed in the research follows a design-science approach. Results: The results of the case study are related to the application of the proposed meta-model to align the specification of a BP model with work practices models. Conclusions: This meta-model can be used within the BPPAM methodology to specify or improve business processes models based on work practice descriptions.

Population studies of D. speciosa on fall / winter crops were conducted. Larvae were monitored on maize (Zea mays L.), wheat (Triticum aestivum L.) and black oats (Avena strigosa Schreb.) and beetles on soybeans (Glycine max (L.) Mill., maize, common beans (Phaseolus vulgaris L.), wheat and black oats from March 23, 2001 to August 24, 2001. Soybean, maize, common beans, wheat and oats were sown on December 28, 2000; February 9, 2001; March 2, 2001; April 26, 2001 and May 11, 2001, respectively. Maize and common beans were grown on latter growing season. Adult beetles of D. speciosa were collected throughout the sampling period. Greatest beetles population peak occurred on wheat in August 3, 2001 which coincided with flowering period. Population dynamics of males and females was similar on common beans and soybeans. Females on maize predominate mostly after the first 30 days after the plant emergence (dae) (before were not detected) until about 45 dae. Males appeared to predominate during the flowering period. Similar population dynamics of males and females were found on wheat and black oats. Greatest peak of larvae occurred on maize roots. The growing season corn farm system was recently introduced, what probably explains the reports of increasing populations of adults during almost the whole year. Probable applications of the results are discussed.

In recent years the wine sector has been characterized by an increased focus on the diversification of wine products. This change has been welcomed by consumers. In fact, today's consumers are willing to experiment with new products and unusual combinationswhich has stimulated research aimed at the discovery of native grape varieties suitable for for winemaking. In this context, particular attention are receiving vines for the production of dessert wines. Many regions have made efforts in the search for varieties and techniques able to better promote the drying process. At CREA-VE Turi-BA, there is an ongoing recovery and exploitation of indigenous grape varieties in the regions of Southern Italy and today more than 3,000 accessions of wine grapes are preserved. In the present work we have investigate the possibility of the use of certain accessions of Vitis vinifera varieties (collection CREA-VE Turi-BA) deemed attractive for the production of dessert wines. Accessions presenting a good sugar content and a good titratable acidity were selected; namely, Greco bianco b. (named Q19) and Mantonico bianco b. (named R8) were compared to the cv of Malvasia di Lipari b. Ampelographic and ampelometric analyses, followed by molecular analysis, were carried out for varietal assessment. A rate of the grapes were fermented immediately after harvest with standard procedure, in comparison to rates vinified after withering. The latter was made in a greenhouse type structure with parts of natural and unforced air, without any additional energy expense. At the end of the process, a loss in weight of the grapes equal to 30–35% was realized. The grapes are then stemmed, crushed, pressed and fermented by selected yeast under controlled temperature. Upon completion of the fermentation and subsequent stabilization, the wines were analyzed for the most important enological parameters and subjected to evaluation by a panel of expert tasters. Chemical analysis of wines showed that an alcohol content ranging to 11–12% volume was reached with in the standard wine-making; whilst, for the vinification of dried grapes the alcohol content was equal to 16% vol for Mantonico bianco b. and 18% vol. for and Greco bianco. Organoleptic analysis of the wines showed a greater appreciation for both of dessert wines. The best ratings were given for the olfactory characteristics (floral and fruity aromas) and taste characteristics. In particular, the Greco bianco b. had excellent scores slightly lower than Malvasia di Lipari b. Therefore, the tested varieties have shown a good potential for the production of dessert wines related to typicality of the grape and the region. Dessert wines produced, have a complex flavor profile and interesting for the high acidic component, which improved the taste perception of wine and attenuated the sensation of sweetness. That feature could allow a good wine aging, too.

Abstract Background Cell surface pili in Gram positive bacteria have been reported to orchestrate the colonization of host tissues, evasion of immunity and the development of biofilms. So far, little if any information is available on the presence of pilus-like structures in human gut commensals like bifidobacteria. Results and discussion In this report, Atomic Force Microscopy (AFM) of various bifidobacterial strains belonging to Bifidobacterium bifidum, Bifidobacterium longum subsp. longum, Bifidobacterium dentium, Bifidobacterium adolescentis and Bifidobacterium animalis subsp. lactis revealed the existence of appendages resembling pilus-like structures. Interestingly, these microorganisms harbour two to six predicted pilus gene clusters in their genome, with each organized in an operon encompassing the major pilin subunit-encoding gene (designated fimA or fimP) together with one or two minor pilin subunit-encoding genes (designated as fimB and/or fimQ), and a gene encoding a sortase enzyme (strA). Quantitative Real Time (qRT)-PCR analysis and RT-PCR experiments revealed a polycistronic mRNA, encompassing the fimA/P and fimB/Q genes, which are differentially expressed upon cultivation of bifidobacteria on various glycans.

Grapevine is one of the most important crop plants in the world. Recently there was great expansion of genomics resources about grapevine genome, thus providing increasing efforts for molecular breeding. Current cultivars display a great level of inter-specific differentiation that needs to be investigated to reach a comprehensive understanding of the genetic basis of phenotypic differences, and to find responsible genes selected by cross breeding programs. While there have been significant advances in resolving the pattern and nature of single nucleotide polymorphisms (SNPs) on plant genomes, few data are available on copy number variation (CNV). Furthermore association between structural variations and phenotypes has been described in only a few cases. We combined high throughput biotechnologies and bioinformatics tools, to reveal the first inter-varietal atlas of structural variation (SV) for the grapevine genome. We sequenced and compared four table grape cultivars with the Pinot noir inbred line PN40024 genome as the reference. We detected roughly 8% of the grapevine genome affected by genomic variations. Taken into account phenotypic differences existing among the studied varieties we performed comparison of SVs among them and the reference and next we performed an in-depth analysis of gene content of polymorphic regions. This allowed us to identify genes showing differences in copy number as putative functional candidates for important traits in grapevine cultivation.

Abstract Background Segmental duplications (SDs) are blocks of genomic sequence of 1-200 kb that map to different loci in a genome and share a sequence identity > 90%. SDs show at the sequence level the same characteristics as other regions of the human genome: they contain both high-copy repeats and gene sequences. SDs play an important role in genome plasticity by creating new genes and modeling genome structure. Although data is plentiful for mammals, not much was known about the representation of SDs in plant genomes. In this regard, we performed a genome-wide analysis of high-identity SDs on the sequenced grapevine (Vitis vinifera) genome (PN40024). Results We demonstrate that recent SDs (> 94% identity and >= 10 kb in size) are a relevant component of the grapevine genome (85 Mb, 17% of the genome sequence). We detected mitochondrial and plastid DNA and genes (10% of gene annotation) in segmentally duplicated regions of the nuclear genome. In particular, the nine highest copy number genes have a copy in either or both organelle genomes. Further we showed that several duplicated genes take part in the biosynthesis of compounds involved in plant response to environmental stress. Conclusions These data show the great influence of SDs and organelle DNA transfers in modeling the Vitis vinifera nuclear DNA structure as well as the impact of SDs in contributing to the adaptive capacity of grapevine and the nutritional content of grape products through genome variation. This study represents a step forward in the full characterization of duplicated genes important for grapevine cultural needs and human health.

Abstract Background Copy number variation (CNV) represents another important source of genetic variation complementary to single nucleotide polymorphism (SNP). High-density SNP array data have been routinely used to detect human CNVs, many of which have significant functional effects on gene expression and human diseases. In the dairy industry, a large quantity of SNP genotyping results are becoming available and can be used for CNV discovery to understand and accelerate genetic improvement for complex traits. Results We performed a systematic analysis of CNV using the Bovine HapMap SNP genotyping data, including 539 animals of 21 modern cattle breeds and 6 outgroups. After correcting genomic waves and considering the pedigree information, we identified 682 candidate CNV regions, which represent 139.8 megabases (~4.60%) of the genome. Selected CNVs were further experimentally validated and we found that copy number "gain" CNVs were predominantly clustered in tandem rather than existing as interspersed duplications. Many CNV regions (~56%) overlap with cattle genes (1,263), which are significantly enriched for immunity, lactation, reproduction and rumination. The overlap of this new dataset and other published CNV studies was less than 40%; however, our discovery of large, high frequency (> 5% of animals surveyed) CNV regions showed 90% agreement with other studies. These results highlight the differences and commonalities between technical platforms. Conclusions We present a comprehensive genomic analysis of cattle CNVs derived from SNP data which will be a valuable genomic variation resource. Combined with SNP detection assays, gene-containing CNV regions may help identify genes undergoing artificial selection in domesticated animals.

Abstract Background Duplicated sequences are an important source of gene innovation and structural variation within mammalian genomes. We performed the first systematic and genome-wide analysis of segmental duplications in the modern domesticated cattle (Bos taurus). Using two distinct computational analyses, we estimated that 3.1% (94.4 Mb) of the bovine genome consists of recently duplicated sequences (≥ 1 kb in length, ≥ 90% sequence identity). Similar to other mammalian draft assemblies, almost half (47% of 94.4 Mb) of these sequences have not been assigned to cattle chromosomes. Results In this study, we provide the first experimental validation large duplications and briefly compared their distribution on two independent bovine genome assemblies using fluorescent in situ hybridization (FISH). Our analyses suggest that the (75-90%) of segmental duplications are organized into local tandem duplication clusters. Along with rodents and carnivores, these results now confidently establish tandem duplications as the most likely mammalian archetypical organization, in contrast to humans and great ape species which show a preponderance of interspersed duplications. A cross-species survey of duplicated genes and gene families indicated that duplication, positive selection and gene conversion have shaped primates, rodents, carnivores and ruminants to different degrees for their speciation and adaptation. We identified that bovine segmental duplications corresponding to genes are significantly enriched for specific biological functions such as immunity, digestion, lactation and reproduction. Conclusion Our results suggest that in most mammalian lineages segmental duplications are organized in a tandem configuration. Segmental duplications remain problematic for genome and assembly and we highlight genic regions that require higher quality sequence characterization. This study provides insights into mammalian genome evolution and generates a valuable resource for cattle genomics research.

Abstract Background Previously published studies have reported that up to 43% of patients with disorders of consciousness are erroneously assigned a diagnosis of vegetative state (VS). However, no recent studies have investigated the accuracy of this grave clinical diagnosis. In this study, we compared consensus-based diagnoses of VS and MCS to those based on a well-established standardized neurobehavioral rating scale, the JFK Coma Recovery Scale-Revised (CRS-R). Methods We prospectively followed 103 patients (55 ± 19 years) with mixed etiologies and compared the clinical consensus diagnosis provided by the physician on the basis of the medical staff's daily observations to diagnoses derived from CRS-R assessments performed by research staff. All patients were assigned a diagnosis of 'VS', 'MCS' or 'uncertain diagnosis.' Results Of the 44 patients diagnosed with VS based on the clinical consensus of the medical team, 18 (41%) were found to be in MCS following standardized assessment with the CRS-R. In the 41 patients with a consensus diagnosis of MCS, 4 (10%) had emerged from MCS, according to the CRS-R. We also found that the majority of patients assigned an uncertain diagnosis by clinical consensus (89%) were in MCS based on CRS-R findings. Conclusion Despite the importance of diagnostic accuracy, the rate of misdiagnosis of VS has not substantially changed in the past 15 years. Standardized neurobehavioral assessment is a more sensitive means of establishing differential diagnosis in patients with disorders of consciousness when compared to diagnoses determined by clinical consensus.

Abstract Background Previous magnetoencephalography (MEG) studies have demonstrated speaking-induced suppression (SIS) in the auditory cortex during vocalization tasks wherein the M100 response to a subject's own speaking is reduced compared to the response when they hear playback of their speech. Results The present MEG study investigated the effects of utterance rapidity and complexity on SIS: The greatest difference between speak and listen M100 amplitudes (i.e., most SIS) was found in the simple speech task. As the utterances became more rapid and complex, SIS was significantly reduced (p = 0.0003). Conclusion These findings are highly consistent with our model of how auditory feedback is processed during speaking, where incoming feedback is compared with an efference-copy derived prediction of expected feedback. Thus, the results provide further insights about how speech motor output is controlled, as well as the computational role of auditory cortex in transforming auditory feedback.

Los modelos media-varianza de selección de cartera, y en general los basados en el binomio rentabilidad-riesgo, parten de un capital inicial cierto. La consideración de la aversión al riesgo en los mismos es indirecta y siempre referida al valor final de la cartera, así que por lo general lo que se determina en los mismos es la proporción de la inversión total o riqueza inicial que se destina a cada activo considerado. Sin embargo, en muchas situaciones reales la aversión al riesgo debe considerarse ya en el momento de invertir, por lo que la determinación de la cuantía a invertir, coste o dotación inicial forma parte del problema, considerando que será mayor, dentro de unas cotas, si existe la probabilidad de obtener mayor rentabilidad aún soportando mayor riesgo. El problema se presenta mediante un modelo simple, pero general en su planteamiento, en el que además de la composición de la cartera queda determinada la cantidad a invertir. La idea es que el inversor establece dos capitales, uno como mínimo y otro como máximo, que está dispuesto a invertir y se utiliza la optimización por escenarios para decidir qué cantidad, entre las dos anteriores, le conviene invertir según sus expectativas.

El objetivo de este trabajo consiste en analizar las características fundamentales de algunos de los paquetes de software comercial que resuelven problemas de programación lineal en el ámbito de la economía. Se han elegido aquéllos más frecuentes en la práctica docente y que resultan fácilmente accesibles para los estudiantes. Se estudia, entre otros, aspectos relacionados con la familiaridad del estudiante con el entorno de trabajo, tipo usual de hardware disponible, documentación y bibliografía disponible, y la aproximación algorítmica utilizada, usando como tests problemas de modelos económicos relevantes.

Durante los últimos años se ha utilizado una nueva metodología en la enseñanza de la investigación operativa en los estudios de Economía y Gestión Empresarial. Los autores imparten docencia de cursos universitarios para estudiantes de dichas disciplinas y creen que el uso del método del caso en las clases prácticas es una forma excelente de satisfacer el deseo de los alumnos de entender el mundo real. En este trabajo se analiza lo que los estudiantes piensan sobre dicha metodología, y en general sobre las clases prácticas, mediante los resultados de una encuesta.

El presente estudio pretende ser una contribución al conocimiento de la cerámica campaniense en el ámbito de la Península Ibérica. En base al material reunido, procedente de diversos yacimientos de la provincia de Sevilla, el planteamiento del trabajo ha girado en torno a la documentación efectiva en dicho ámbito provincial tanto de la cerámica campaniense de tipo C como de una serie de cerámicas caracterizadas básicamente por su pasta gris y que en ocasiones, por sus características técnicas, formales o decorativas apuntan a una inspiración más o menos directa en las producciones propiamente campanienses.

Introduction The US–Mexico border region has 15 million residents and 300,000 births annually. Reproductive health concerns have been identified on both sides of the border, but comparable information about reproductive health is not available. The objective of this study was to compare reproductive health indicators among populations in this region. Methods We used 2009 US Hispanic and Mexican birth certificate data to compare births inside the border region, elsewhere within the border states, and in the United States and Mexico overall. We examined trends in total fertility and birth rates using birth data from 2000 through 2009 and intercensal population estimates. Results Among women in the border region, US women had more lifetime births than Mexican women in 2009 (2.69 births vs 2.15 births) and throughout the decade. Birth rates in the group aged 15 to 19 years were high in both the US (73.8/1,000) and Mexican (86.7/1,000) border regions. Late or no prenatal care was nearly twice as prevalent in the border regions as in the nonborder regions of border states. Low birth weight and preterm and early-term birth were more prevalent in the US border than in the Mexican border region; US border rates were higher and Mexican rates were lower than their corresponding nonborder and national rates. We found some variations within border states. Conclusion These findings constitute the first population-based information on the reproductive health of the entire Hispanic US–Mexico border population. Evidence of disparities warrants exploration at state and local levels. Teen pregnancy and inadequate prenatal care are shared problems in US–Mexico border communities and suggest an area for binational cooperation.

Assessment of forest ecosystems and their services is seen as a key action for the advancement of biodiversity objectives, and to inform the development and implementation of related policies and planning. The Azorean forest is important for timber production, the protection of soil and water resources, and for its recreational and aesthetic value. However, its role in carbon accumulation has not been fully addressed. We assessed plant diversity, forest structure and carbon stocks in a gradient of three forest types (Natural Forest-NF; Exotic Woodland-EW and Production Forest-PF) in three of the Azores islands. We used biodiversity indices and found that NF harbored the highest plant diversity levels and PF the lowest. Diversity levels were lower for structural than for taxonomic data, particularly for PF. The highest tree carbon stock was found at EW in one of the islands, while PF consistently exhibited relatively high tree carbon stocks in the three islands. The largest soil carbon stocks were found at EW, while leaf litter carbon stocks were higher at PF. We concluded that NF play a fundamental role as plant diversity hotspots but have lower relevance as carbon stocks what might be associated with montane environmental conditions. PFs provide economic assets and act as carbon sinks, while EWs play a major role as carbon sinks in soil, but also at tree level in the oldest forests., (© 2022. The Author(s).)

Background: Saliva is a complex secretion produced daily by the salivary glands. Saliva consists mainly of water, enzymes, ions and amino acids and performs several important functions in oral health., Objective: The aim of this study was to investigate the flow rate and concentrations of amylase and total proteins in the saliva of hospitalized patients due to AIDS complications., Methods: Ninety-three men and women (20-64 years of age) were divided into two groups (46 HIV-infected patients and 47 controls) and had salivary flow rate and levels of amylase enzyme and total proteins evaluated., Result: The mean salivary flow rate was lower in individuals with HIV when compared to controls (P < 0.05). No significant difference between amylase enzyme levels and total proteins were observed in the saliva of patients with HIV infection when compared to controls., Conclusion: Individuals with HIV / AIDS infection (in hospital treatment) suffer no interference in levels of amylase and total salivary proteins, but they have significantly reduced salivary flow., (© 2020 Carina de Oliveira N et al.)

The signaling pathway of G protein-coupled receptors is strongly linked to their trafficking profile. Little is known about the molecular mechanisms involved in the vasopressin receptor V 1b subtype (V 1b R) trafficking and its impact on receptor signaling and regulation. For this purpose, we investigated the role of β-arrestins in receptor desensitization, internalization and recycling and attempted to dissect the V 1b R-mediated MAP kinase pathway. Using MEF cells Knocked-out for β-arrestins 1 and 2, we demonstrated that both β-arrestins 1 and 2 play a fundamental role in internalization and recycling of V 1b R with a rapid and transient V 1b R-β-arrestin interaction in contrast to a slow and long-lasting β-arrestin recruitment of the V 2 vasopressin receptor subtype (V 2 R). Using V 1b R-V 2 R chimeras and V 1b R C-terminus truncations, we demonstrated the critical role of the V 1b R C-terminus in its interaction with β-arrestins thereby regulating the receptor internalization and recycling kinetics in a phosphorylation-independent manner. In parallel, V 1b R MAP kinase activation was dependent on arrestins and Src-kinase but independent on G proteins. Interestingly, Src interacted with hV 1b R at basal state and dissociated when receptor internalization occurred. Altogether, our data describe for the first time the trafficking profile and MAP kinase pathway of V 1b R involving both arrestins and Src kinase family., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Voriconazole (VCZ) exhibits great inter- and intrapatient variability. The latter variation cannot exclusively be explained by concomitant medications, liver disease or dysfunction, and genetic polymorphisms in cytochrome P450 2C19 (CYP2C19). We hypothesized that inflammatory response in patients under VCZ medication might also influence this fluctuation in concentrations. In this study, we explored the association between inflammation, reflected by the C-reactive protein (CRP) concentration, and VCZ trough concentrations over time. A retrospective analysis of data was performed for patients with more than one steady-state VCZ trough concentration and a CRP concentration measured on the same day. A longitudinal analysis was used for series of observations obtained from many study participants over time. The approach involved inclusion of random effects and autocorrelation in linear models to reflect within-person cross-time correlation. A total of 50 patients were eligible for the study, resulting in 139 observations (paired VCZ and CRP concentrations) for the analysis, ranging from 2 to 6 observations per study participant. Inflammation, marked by the CRP concentration, had a significant association with VCZ trough concentrations (P < 0.001). Covariates such as age and interacting comedication ([es]omeprazole), also showed a significant correlation between VCZ and CRP concentrations (P < 0.05). The intrapatient variation of trough concentrations of VCZ was 1.401 (confidence interval [CI], 0.881 to 2.567), and the interpatient variation was 1.756 (CI, 0.934 to 4.440). The autocorrelation between VCZ trough concentrations at two sequential time points was calculated at 0.71 (CI, 0.51 to 0.92). The inflammatory response appears to play a significant role in the largely unpredictable pharmacokinetics of VCZ, especially in patients with high inflammatory response, as reflected by high CRP concentrations., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Voriconazole pharmacokinetics shows a large inter- and intrapatient variability. Inflammation is associated with changes in the expression of CYP isoenzymes. Here, we evaluated the influence of inflammation, marked by C-reactive protein (CRP) levels in blood, on the metabolism of voriconazole. Observational data showed an association between CRP level and the ratio of voriconazole N-oxide to voriconazole., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Objective: To assess the technique employed by the autoanalyser Architect® i1000sr to determine digoxin in serum samples, compared with the assay developed for AsSYM® using microparticle enzyme immunoassay (DigoxinII)., Method: A prospective analysis of the samples from 100 requests to monitor patients being treated with digoxin. The samples were processed in AxSYM® and Architect®. The techniques were assessed using the linear regression coefficient, determination coefficient, mean absolute error, mean squared prediction error and the Bland-Altman method., Results: The serum levels showed a correlation coefficient of 0.93. There was nearly a 40% difference for the concentrations between 0.8 and 2 ng/ml and nearly 20% in the rest of the samples analysed., Conclusions: The Architect® system is precise; however, from a clinical monitoring point of view, it is unacceptably inaccurate when compared with the AxSYM®., (Copyright © 2010 SEFH. Published by Elsevier Espana. All rights reserved.)

Recently, we synthesized and characterized the first selective V(1b) vasopressin (VP)/oxytocin receptor agonist, d[Cha(4)]arginine vasopressin. However, this agonist was only selective for the human receptors. We thus decided to design a selective V(1b) agonist for the rodent species. We started from previous observations showing that modifying [deamino(1),Arg(8)]VP in positions 4 and 8 altered the rat VP/oxytocin receptor selectivity. We synthesized a series of 13 [deamino(1),Arg(8)]VP analogs modified in positions 4 and 8. Among them, one seemed very promising, d[Leu(4), Lys(8)]VP. In this paper, we describe its pharmacological and physiological properties. This analog exhibited a nanomolar affinity for the rat, human, and mouse V(1b) VP receptors and a strong V(1b) selectivity for the rat species. On AtT20 cells stably transfected with the rat V(1b) receptor, d[Leu(4), Lys(8)]VP behaved as a full agonist on both phospholipase C and MAPK assays. Additional experiments revealed its ability to induce the internalization of enhanced green fluorescent protein-tagged human and mouse V(1b) receptors as expected for a full agonist. Additional physiological experiments were performed to further confirm the selectivity of this peptide. Its antidiuretic, vasopressor, and in vitro oxytocic activities were weak compared with those of VP. In contrast, used at low doses, its efficiency to stimulate adrenocorticotropin or insulin release from mouse pituitary or perfused rat pancreas, respectively, was similar to that obtained with VP. In conclusion, d[Leu(4), Lys(8)]VP is the first selective agonist available for the rat V(1b) VP receptor. It will allow a better understanding of V(1b) receptor-mediated effects in rodents.

[(3)H]SSR-149415 is the first tritiated nonpeptide vasopressin V(1b) receptor (V(1b)R) antagonist ligand. It was used for studying rodent (mouse, rat, hamster) and human V(1b)R from native or recombinant origin. Moreover, a close comparison between the human and the mouse V(1b)R was performed using SSR-149415/[(3)H]SSR-149415 in binding and functional studies in vitro. [(3)H]SSR-149415 binding was time-dependent, reversible, and saturable. Scatchard plot analysis gave a single class of high-affinity binding sites with apparent equilibrium dissociation constant (K(d)) approximately 1 nM and maximum binding density (B(max)) values from 7,000 to 300,000 sites/cell according to the cell line. In competition experiments, [(3)H]SSR-149415 binding was stereospecific and dose-dependently displaced by reference peptide and nonpeptide arginine vasopressin (AVP)/OT ligands following a V(1b) rank order of affinity: SSR-149415 = AVP > dCha > dPen > dPal > dDavp > SSR-126768A > SR-49059 > SSR-149424 > OT > SR-121463B. Species differences between human, rat, mouse, and hamster V(1b)R were observed. Autoradiography studies with [(3)H]SSR-149415 on rat and human pituitary showed intense specific labeling confined to corticotroph cells and absence of labeling in the other tissues examined. SSR-149415 potently and stereospecifically antagonized the AVP-induced inositol phosphate production and intracellular Ca(2+) increase (EC(50) from 1.83 to 3.05 nM) in recombinant cell lines expressing either the mouse or the human V(1b)R. AVP (10(-7) M) exposure of AtT20 cells expressing mouse or human EGFP-tagged V(1b)R induced their rapid internalization. Preincubation with 10(-6) M SSR-149415 counteracted the internalization process. Moreover, recycling of internalized receptors was observed upon 10(-6) M SSR-149415 treatment. Thus SSR-149415/[(3)H]SSR-149415 are unique tools for studying animal and human V(1b)R.

Cell-surface expression and biological functions of several intracellular-retained G protein-coupled receptors are restored by membrane-permeable ligands called pharmacological chaperones. We have previously demonstrated that a mutation of the hydrophobic motif 341FNX2LLX3L350 in the C terminus of the human pituitary vasopressin V3 receptor (MUT V3R) led to it being retained in the endoplasmic reticulum (ER). Here, we establish the precise role of this motif and investigate whether SSR149415, a non-peptide V3R antagonist, behaves as a pharmacological chaperone for the ER-retained MUT V3R. The absence of the mutated receptor in the plasma membrane is linked to its prolonged association with the molecular chaperone calnexin in the ER and to its intensive degradation by the ubiquitin-proteasomal machinery. However, this is not because of a lack of oligomerization, as demonstrated by the presence of MUT V3R homodimers in the ER. Treatment with SSR149415 restores expression of the mutated receptor on the cell surface and its correct maturation, resulting into the functional recovery of its signaling properties. SSR149415 acts by stabilizing a native-like conformation of the V3R, reducing its association with calnexin and, thus, favoring a secretory pathway rather than the proteasomal degradation pathway. In conclusion, the FN(X)2LL(X)3L sequence is an important motif for the V3R conformation, and the misfolding resulting from its mutation alters the receptor export but can be reverted by SSR149415.

Little is known about endoplasmic reticulum (ER) export signals, particularly those of members of the G-protein-coupled receptor family. We investigated the structural motifs involved in membrane export of the human pituitary vasopressin V1b/V3 receptor. A series of V3 receptors carrying deletions and point mutations were expressed in AtT20 corticotroph cells. We analyzed the export of these receptors by monitoring radioligand binding and by analysis of a V3 receptor tagged with both green fluorescent protein and Myc epitopes by a novel flow cytometry-based method. This novel method allowed us to quantify total and membrane-bound receptor expression. Receptors lacking the C terminus were not expressed at the cell surface, suggesting the presence of an export motif in this domain. The distal C terminus contains two di-acidic (DXE) ER export motifs; however, mutating both these motifs had no effect on the V3 receptor export. The proximal C terminus contains a di-leucine (345)LL(346) motif surrounded by the hydrophobic residues Phe(341), Asn(342), and Leu(350). The mutation of one or more of these five residues abolished up to 100% of the receptor export. In addition, these mutants colocalized with calnexin, demonstrating that they were retained in the ER. Finally, this motif was sufficient to confer export properties on a CD8alpha glycoprotein-V3 receptor chimera. In conclusion, we have identified a novel export motif, FN(X)(2)LL(X)(3)L, in the C terminus of the V3 receptor.

In the pituitary, vasopressin triggers ACTH release through a specific receptor subtype, termed V3 or V1b. We cloned the V3 cDNA and showed that its expression was almost exclusive to pituitary corticotrophs and some corticotroph tumors. To study the determinants of this tissue specificity, we have now cloned the gene for the human (h) V3 receptor and characterized its structure. It is composed of two exons, spanning 10kb, with the coding region interrupted between transmembrane domains 6 and 7. We established that the transcription initiation site is located 498 nucleotides upstream of the initiator codon and showed that two polyadenylation sites may be used, while the most frequent is the most downstream. Sequence analysis of the promoter region showed no TATA box but identified consensus binding motifs for Sp1, CREB, and half sites of the estrogen receptor binding site. However comparison with another corticotroph-specific gene, proopiomelanocortin, did not identify common regulatory elements in the two promoters except for a short GC-rich region. Unexpectedly, hV3 gene analysis revealed that a formerly cloned 'artifactual' hV3 cDNA indeed corresponded to a spliced antisense transcript, overlapping the 5' part of the coding sequence in exon 1 and the promoter region. This transcript, hV3rev, was detected in normal pituitary and in many corticotroph tumors expressing hV3 sense mRNA and may therefore play a role in hV3 gene expression.

Ten patients with high-grade non-Hodgkin's lymphoma (HG-NHL) entered a subcutaneous (s.c.) recombinant interleukin 2 (rIL2) trial within 2 months of undergoing autologous bone marrow transplantation (ABMT). Immunological studies, consisting in T- and natural killer (NK)-cell subset assessment, together with functional assays, such as NK activity and CD16-mediated redirected killing assay, were performed before therapy, after 2 weeks, and then monthly. Phenotypic analysis showed a significant increase (p = 0.01) of CD16 and CD56 NK cells, from 12% to 28% and from 17% to 37%, respectively. In particular, the CD56bright NK cell population showed a tenfold increase, while CD56dim NK cells remained unmodified compared with pretreatment values. The expression of IL2 receptors was also studied and a significant increase (p = 0.01) of CD122 (p75)-positive cells from 8% to 30% was found, while no significant increase was observed in CD25 (p55)-positive cells. Furthermore, rIL2 administration led to an increase of NK activity even at the lowest effectors:target ratio and to an increase of CD16-mediated redirected killing assay. These phenotypic and functional modifications lasted throughout the duration of rIL2 therapy and remained after completion of therapy. In addition, none of the ten patients relapsed, and two of them who started IL2 treatment while still showing residual disease experienced a complete disappearance of the disease after 10 and 7 months of therapy, respectively. Our data suggest that infusion of rIL2 s.c. after ABMT is safe, can selectively increase NK cell number and function, and may have a beneficial effect on the minimal residual disease.

The immunophenotype of 110 adult patients with diagnosis of acute myeloblastic leukemia (AML) was analyzed using a wide panel of monoclonal antibodies (mAbs). Leukemic blasts were tested by applying direct immunofluorescence analysis and dual-fluorescence staining, and two groups of patients were identified: 56/110 (51%) expressing only myeloid antigens (My/AML) and 54/110 (49%) expressing both myeloid and lymphoid antigens (Ly/AML). CD13 and CD33 were expressed in almost all FAB subtypes, whereas CD14, frequently expressed in M4 and M5 subtypes (70%), was rarely expressed in M0 + M1 cases (9%). On the contrary, CD34, expressed in 77% of M0 + M1 cases, was practically absent in M3 and M5 subtypes (6% and 7%, respectively). CD2 and CD7 antigens were found in 34% and 42% of patients respectively, whereas B cell-associated antigens, such as CD10 and CD19, were found in 31% and 18% of patients. Cytogenetic abnormalities characteristically present in AML patients were also analyzed and, except for t(8;21) which was found in both groups of patients, the other abnormalities were frequently found in cases coexpressing lymphoid-associated antigens. Finally, the complete remission (CR) rate, survival and event-free survival were analyzed according to the presence of lymphoid markers and also of some specific antigens such as CD7 and CD34. The only prognostic difference was represented by CD34+ patients who showed a reduction in the CR rate compared with CD34- patients (65% versus 82%) (p = 0.05) which became more evident when the mean intensity of fluorescence was considered.(ABSTRACT TRUNCATED AT 250 WORDS)