Anthocyanin-Loaded Polymers as Promising Nature-Based, Responsive, and Bioactive Materials.

Anthocyanins are a specific group of molecules found in nature that have recently received increasing attention due to their interesting biological and colorimetric properties that have been successfully applied in several fields such as food preservation and biomedicine. Consequently, reviews devoted to a general overview of these flavonoids have proliferated in recent years. Meanwhile, the incorporation of anthocyanins into polymeric systems has become an interesting strategy to widen the applicability of these molecules and develop new smart and functional polymers in the above cited areas. However, anthocyanin-based polymers have been scarcely reviewed in the literature. Accordingly, this review aims to be a systematic summary of the most recent approaches for the incorporation of anthocyanins into macro-, micro-, or nanostructured polymers. Moreover, this work describes the fundamentals of the applicability of smart anthocyanin-based polymers and offers an updated review of their most interesting applications as sensors, biological regulators, and active materials.

Air pollution and surrounding greenness in relation to ischemic stroke: A population-based cohort study.

BACKGROUND: Evidence for the association between environmental exposures and ischemic stroke (IS) is limited and inconsistent. We aimed to assess the relationship between exposure to air pollutants, residential surrounding greenness, and incident IS, and to identify population subgroups particularly sensitive to these exposures. METHODS: We used data from administrative health registries of the public healthcare system in Catalonia, Spain to construct a cohort of individuals aged 18 years and older without a previous stroke diagnosis at 1st January 2016 (n = 3 521 274). We collected data on sociodemographic characteristics and cerebrovascular risk factors, and derived exposure at the participant's residence to ambient levels of fine particulate matter (PM2.5), black carbon (BC), nitrogen dioxide (NO2), and Normalized Difference Vegetation Index (NDVI) in a 300 m buffer as an indicator of greenness. The primary outcome was IS diagnosis at any point during the follow-up. We used Cox proportional hazards models to estimate associations between environmental exposures and incident IS and stratified analyses to investigate effect modification. RESULTS: Between 1st January 2016 and 31st December 2017, 10 865 individuals were admitted to public hospitals with an IS diagnosis. Median exposure levels were: 17 µg/m3 PM2.5, 35 µg/m3 NO2, 2.28 µg/m3 BC and 0.27 NDVI. Individuals with higher residential exposure to air pollution were at greater risk of IS: HR 1·04 (95% CI:0·99-1·10) per 5 µg/m3 of PM2.5; HR 1.05 (95% CI:1·00-1·10) per 1 µg/m3 of BC; HR 1·04 (95% CI:1·03-1·06) per 10 µg/m3 of NO2. Conversely, individuals with higher residential surrounding green space, had lower risk of IS (HR 0·84; CI 95%:0·7-1.0). There was no evidence of effect modification by individual characteristics. CONCLUSIONS: Higher incidence of IS was observed in relation to long-term exposures to air pollution, particularly NO2, in a region that meets European health-based air quality standards. Residential surrounding greenness was associated with lower incidence of IS.

The Mexican consensus on the diagnosis, treatment, and prevention of NSAID-induced gastropathy and enteropathy. / Consenso mexicano sobre diagnóstico, prevención y tratamiento de la gastropatía y enteropatía por antiinflamatorios no esteroideos.

More than 30 million persons worldwide take nonsteroidal anti-inflammatory drugs (NSAIDs) on a daily basis, and annual consumption is increasing. In addition to their analgesic and anti-inflammatory properties, NSAIDs also produce well-known gastrointestinal adverse events. There is no consensus in Mexico on the diagnosis, treatment, and prevention of NSAID-induced gastropathy and enteropathy, and so the Asociación Mexicana de Gastroenterología brought together a group of experts to establish useful recommendations for the medical community. Thirty-three recommendations were formulated in the present consensus, highlighting the fact that the risk for NSAID-induced gastrointestinal toxicity varies according to the drug employed and its pharmacokinetics, which should be taken into account at the time of prescription. The risk factors for gastroduodenal complications due to NSAIDs are: a history of peptic ulcer, age above 65 years, high doses of NSAIDs, Helicobacter pylori infection, and the presence of severe comorbidities. The symptoms and gastroduodenal damage induced by NSAIDs vary, ranging from an asymptomatic course to the presentation of iron-deficiency anemia, bleeding, stricture, and perforation. Capsule endoscopy and enteroscopy are direct diagnostic methods in NSAID enteropathy. Regarding prevention, the minimum dose of an NSAID needed to achieve the desired effect, administered for the shortest period of time, is the recommendation. Finally, proton pump inhibitors are the gold standard for the prophylaxis and treatment of gastroduodenal effects, but they are not useful in enteropathy.

Oxidative stress modulation induced by chitosan-glutathione nanoparticles in chondrocytes.

The use of nanometric systems to deliver biologically active substances is a successful tool in different fields. In this study, we investigated nanometric systems with antioxidant capacity to modulate events associated with the redox state in human chondrocytes. We used nanoparticles (NPs) prepared with chitosan and glutathione (GSH) and an in vitro model: primary cultures of human chondrocytes were extracted from hyaline cartilage. The cells were exposed to CdCl2 in the presence or absence of NPs. CdCl2 is a widely known oxidizing agent. Fluorescence and confocal microscopy showed the location of the NPs within the cells. The results obtained showed that the NPs did not significantly affect cell viability. We studied the antioxidant capacity of the NPs by estimating the GSH, TBARs, and Cell Rox content and the enzymatic activity of glutathione peroxidase (GPx). In vitro assays showed that GSH levels, GPx activity and reactive oxygen species (Cell Rox) levels were modified with both concentrations of NPs, while lipoperoxidation (TBARs) decreased when cells exposed to CdCl2 were in contact with the NPs. All these results suggest the ability of NPs to modulate the cell redox state in a dose-dependent manner.

Brain regions related to fear extinction in obsessive-compulsive disorder and its relation to exposure therapy outcome: a morphometric study.

BACKGROUND: The size of particular sub-regions within the ventromedial prefrontal cortex (vmPFC) has been associated with fear extinction in humans. Exposure therapy is a form of extinction learning widely used in the treatment of obsessive-compulsive disorder (OCD). Here we investigated the relationship between morphometric measurements of different sub-regions of the vmPFC and exposure therapy outcome in OCD. METHOD: A total of 74 OCD patients and 86 healthy controls underwent magnetic resonance imaging (MRI). Cortical thickness and volumetric measurements were obtained for the rostral anterior cingulate cortex (rACC), the medial orbital frontal cortex and the subcallosal cortex. After MRI acquisition, patients were enrolled in an exposure therapy protocol, and we assessed the relationship between MRI-derived measurements and treatment outcome. Baseline between-group differences for such measurements were also assessed. RESULTS: Compared with healthy controls, OCD patients showed a thinner left rACC (p = 0.008). Also, left rACC thickness was inversely associated with exposure therapy outcome (r - 0.32, p = 0.008), and this region was significantly thinner in OCD patients who responded to exposure therapy than in those who did not (p = 0.006). Analyses based on regional volumetry did not yield any significant results. CONCLUSIONS: OCD patients showed cortical thickness reductions in the left rACC, and these alterations were related to exposure therapy outcome. The precise characterization of neuroimaging predictors of treatment response derived from the study of the brain areas involved in fear extinction may optimize exposure therapy planning in OCD and other anxiety disorders.

Trophic transfer of pyrene metabolites between aquatic invertebrates.

The trophic transfer of pyrene metabolites was studied using Gammarus setosus as a predator and the invertebrates Lumbriculus variegatus and Chironomus riparius as prey. The results obtained by liquid scintillation counting confirmed that the pyrene metabolites produced by the aquatic invertebrates L. variegatus and C. riparius were transferred to G. setosus through the diet. More detailed analyses by liquid chromatography discovered that two of the metabolites produced by C. riparius appeared in the chromatograms of G. setosus tissue extracts, proving their trophic transfer. These metabolites were not present in chromatograms of G. setosus exclusively exposed to pyrene. The present study supports the trophic transfer of PAH metabolites between benthic macroinvertebrates and common species of an arctic amphipod. As some PAH metabolites are more toxic than the parent compounds, the present study raises concerns about the consequences of their trophic transfer and the fate and effects of PAHs in natural environments.

Efecto nutracéutico del Anacardium occidentale en dietas de pollitas ponedoras de reemplazo / Nutraceutical effect of Anacardium occidentale in diets of replacement laying pullets

Objetivo. Evaluar el efecto nutracéutico del polvo de hojas y retoños de Anacardium occidentale (AO) en dietas de pollitas ponedoras de remplazo. Materiales y métodos. Se utilizaron 240 pollitas White Leghorn (L-33) de un día de edad, que se ubicaron durante 35 días, según diseño completamente aleatorizado, con niveles de adición de 0, 0.5, 1.5 y 2.5% de polvo de hojas y retoños de Anacardium occidentale en las dietas. Se determinaron en las pollitas, los indicadores productivos, peso absoluto y relativo de los órganos inmunes, vísceras, accesorios e intestinos, la hipersensibilidad intestinal y la glucosa sérica. Resultados. El peso vivo final, consumo de alimento, peso del timo, bolsa de Fabricio y colon + recto en las aves con el tracto gastrointestinal vacío y lleno, fue favorable con la adición de 0.5% de polvo AO, con diferencias significativas (p≤0.05). El consumo acumulado, el consumo de polvo AO y taninos se incrementaron en las aves con la adición de 1.5 y 2.5% de polvo AO con respecto al control; no obstante los indicadores productivos para estos animales se deprimieron. La adición del polvo de AO, no deterioró el peso relativo de las vísceras (corazón, hígado y riñón) en las aves, además redujo la hipersensibilidad intestinal y la glucosa sérica. Conclusiones. La adición de 0.5% de polvo de hojas y retoños de AO como nutracéutico en las dietas de pollitas ponedoras de remplazo, mejoró los indicadores productivos y el peso de los órganos inmunes; además, la adición del polvo AO en las dietas disminuyó la hipersensibilidad intestinal y la glucosa sérica.

Objective. To assess the nutraceutical effect of powder from leaves and shoots of Anacardium occidentale (AO) in the diets of replacement laying pullets. Materials and methods. 240 day-old White Leghorn chicks (L-33), were placed for 35 days, according to a completely randomized design with addition levels of 0, 0.5, 1.5 and 2.5% of leaves and shoots powder of AO, in their diets. In the case of chicks, production indicators, absolute and relative weight of immune organs, viscera, innards and intestines, intestinal hypersensitivity and serum glucose were determined. Results. The final body weight, the cumulative feed intake, the weight of thymus, bursa of Fabricio, and colon + rectum with empty and full gastrointestinal tract in the birds, were favorable with the addition of 0.5 % of AO powder with significant difference between (p≤0.05). Cumulative intake, consumption of AO powder and tannins were increased in birds with 1.5 and 2.5% of AO powder compared to the control group; however, the production indicators for these animals were reduced. The addition of AO powder did not impair the relative weight of the viscera (heart, liver and kidney) in birds; also, the intestinal hypersensitivity and serum glucose were reduced. Conclusions. The addition of 0.5% of powder from leaves and shoots of AO as nutraceutical in diets for replacement pullets and laying hens, improved the productive indicators and weight of immune organs, plus the addition of three levels of dust in the diets, decreased intestinal hypersensitivity and serum glucose.

Involvement of AMPA/kainate-excitotoxicity in MK801-induced neuronal death in the retrosplenial cortex.

MK801 is a prototypical non-competitive NMDA receptor-antagonist that induces behavioural changes and reversible toxicity at low doses, while at higher doses triggers neuronal death that mainly affects the retrosplenial cortex (RSC) and to a lesser extent other structures such as the posterolateral cortical amygdaloid nucleus (PLCo). The mechanism of MK801-induced neurodegeneration remains poorly understood. In this study we analysed the participation of GABA-ergic and glutamatergic neurotransmission in MK801-induced neuronal death. We used a single i.p. injection of MK801 (2.5 mg/kg) that induced moderate neuronal death in the RSC and PLCo of female rats, and combined this treatment with the i.p., i.c.v., or intra-RSC infusion of drugs that are selective agonists or antagonists of the GABA-ergic or glutamatergic neurotransmission. We found that neuronal death in the RSC, but not the PLCo, was significantly reduced by the i.p. injection of thiopental, and the i.c.v. application of muscimol, both GABA-A agonists. MK801-toxicity in RSC was abrogated by intra-RSC infusion of muscimol, but the GABA antagonist picrotoxin had no effect. HPLC-analysis showed that levels of glutamate, but not GABA, in the RSC decreased after i.p. treatment with MK801. Intra-RSC infusion of MK801 did not enhance toxicity triggered by the i.p. injection of MK801, indicating that toxicity is not due to direct blockade of NMDA receptors in RSC neurons. MK801-toxicity in the RSC was abrogated by i.c.v. and intra-RSC infusions of the AMPA/kainate antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX). Interestingly, i.c.v. application of neither muscimol or DNQX inhibited MK801-toxicity in the PLCo, suggesting that the mechanism of neuronal death in the RSC and the PLCo might be different. 1-naphthylacetyl spermine trihydrochloride (NASPM), which blocks Ca2+ permeable AMPA/kainate receptors, also reduced MK801-induced toxicity in the RSC. Intra-RSC infusion of AMPA or kainic acid alone promoted death of RSC neurons and was reminiscent of the degeneration induced by the i.p. treatment with MK801. Collectively, these experiments provide evidence for an AMPA/kainate-dependent mechanism of excitotoxicity in the death of RSC neurons after i.p. treatment with MK801.

Clinical and Radiological Differences in Alzheimer's Disease with and without a History of Mild Cognitive Impairment.

We assessed the influence of a history of amnesic mild cognitive impairment (MCI) in patients with Alzheimer's disease (AD) at presentation from a clinical and radiological point of view. A consecutive sample of patients fulfilling the criteria of probable Alzheimer's disease according to the NINCDS-ADRDA work group not previously diagnosed nor treated underwent neuropsychological assessment including mini-mental test, Blessed dementia rating scale (BDRS), ADAS-Cog, neuropsychiatric inventory (NPI), and a daily living activities scale (DAD). Radiological evaluation consisted of magnetic resonance imaging, left hippocampal volumetry and magnetic resonance spectroscopy (MRS) of the brain. We divided the patients into two groups according to positive/negative history of MCI for a period longer than one year. The AD patients with a history of MCI were more likely to have a history of depression (OR: 5.5; 95% CI: 1.12-26) and have more depressive symptoms at presentation in the NPI than those without a history of MCI. They also had a history of hypertension more frequently than the remainder. The presence/absence of ApoE4 alleles did not have influence in the clinical course. With regard to radiological findings the patients with previous MCI showed lower values of N-acetyl-aspartate (NAA) in parietal (p=0.0001) temporal (p=0.08) and occipital (p=0.00001) lobes than the other group, as well as a smaller left hippocampus although the difference was not statistically significant. A history of MCI in AD patients represents a form of the disease with slower progression from clinical and radiological viewpoints. These patients present with more depressive symptoms and a history of depression than the remainder. The lower NAA levels on MRS are compatible with a longer disease duration when AD is preceded by amnesic MCI.

Neurodegeneration and prolonged immediate early gene expression throughout cortical areas of the rat brain following acute administration of dizocilpine.

N-methyl-d-aspartate receptor antagonist drugs (NMDA-A), such as dizocilpine (MK801), induce long-lasting behavioral disturbances reminiscent to psychotic disorders in humans. To identify cortical structures affected by NMDA-A, we used a single dose of MK801 (10 mg/kg) that caused low and high neurodegeneration in intact and orchiectomized male rats, respectively. Degenerating somas (neuronal death) and axonal/synaptic endings (terminal degeneration) were depicted by a silver technique, and functionally affected cortical neuronal subpopulations by Egr-1, c-Fos, and FosB/DeltaFosB-immunolabeling. In intact males, MK801 triggered a c-Fos induction that remained high for more than 24 h in selected layers of the retrosplenial, somatosensory and entorhinal cortices. MK801-induced neurodegeneration reached its peak at 72 h. Degenerating somas were restricted to layer IV of the granular subdivision of the retrosplenial cortex, and were accompanied by suppression of Egr-1 immunolabeling. Terminal degeneration extended to selected layers of the retrosplenial, somatosensory and parahippocampal cortices, which are target areas of retrosplenial cortex. Induction of FosB/DeltaFosB by MK801 also extended to the same cortical layers affected by terminal degeneration, likely reflecting the damage of synaptic connectivity. In orchiectomized males, the neurodegenerative and functional effects of MK801 were exacerbated. Degenerative somas in layer IV of the retrosplenial cortex significantly increased, with a parallel enhancement of terminal degeneration and FosB/DeltaFosB-expression in the mentioned cortical structures, but no additional areas were affected. These observations reveal that synaptic dysfunction/degeneration in the retrosplenial, somatosensory and parahippocampal cortices might underlie the long-lasting impairments induced by NMDA-A.

Increased rewarding properties of morphine in perinatally protein-malnourished rats.

In the current research, we assessed the influence of a protein malnutrition schedule from the 14th day of gestation up to 40 days of age (D-rats) on the rewarding properties of morphine in adult rats by means of the conditioned place preference paradigm. Well-nourished animals (C-rats) administered with different doses of morphine (0.75, 1.5, 3, 6, 12 or 24 mg/kg i.p.) exhibited a conditioning place preference with doses of 3 and 6 mg/kg, whereas in D-rats such a conditioning effect was observed with doses of 1.5 and 3 mg/kg. No adverse effects were observed in either C- or D-rats for the higher doses of morphine. In addition, when animals of both groups were pretreated twice a day for 3 days with increasing doses of morphine (5, 10 and 20 mg/kg s.c.), only D-rats elicited sensitization to the conditioning effect with the lowest dose of morphine (0.75 mg/kg i.p.). Furthermore, sensitized D-rats showed a selective and significant increase in FosB expression in the nucleus accumbens (core and shell), basolateral amygdala and medial prefrontal cortex, brain areas that are functionally related to the rewarding neural circuit. These results demonstrate that a deficient nutritional status during the perinatal period results in adult subjects having neural alterations, leading to an increased responsiveness to morphine and/or enhanced reinforcement effects, which correlates with an overexpression of FosB in selective brain areas related to the rewarding network.

Effects of mid-summer transport duration on pre- and post-slaughter performance and pork quality in Mexico.

Seven hundred and fourteen pigs were monitored from transport to slaughter in July in three treatments: 8, 16 and 24 transport hours; lairage time for the three groups was of 8h. Transport duration significantly (P<0.05) affected live-weight gain during the rest period. Weight gain percentages at lairage were 0.05%, 0.78% and 1.15% for treatments 1, 2 and 3, respectively. Transport to slaughter loss percentage was 2.7%, 4.3% and 6.8% for each of the treatments. Short transport periods significantly increased carcass pH below normal values. Animals transported under acute stress (8h) showed pale carcasses (high possibilities of transforming into PSE meat). On the contrary, pigs transported for 24h had more dark red carcasses. Transport from farm to the slaughterhouse should take no more than 16h in order to improve carcass quality and animals' welfare.

Consultas a un Centro de Información de Medicamentos sobre Atención Farmacéutica / No disponible

Introducción: Un CIM está destinado a proporcionar información para promover el uso racional de medicamentos. Su principal actividad es resolver consultas. Los usuarios de los CIM colegiales son fundamentalmente farmacéuticos comunitarios. Éstos deben dirigir su práctica a la Atención Farmacéutica: dispensación, consulta farmacéutica, seguimiento farmacoterapéutico..., actividades orientadas al paciente y relacionadas con el uso racional del medicamento. Hay, pues, una gran coincidencia entre el propósito de los CIM y el de estas actividades. Luego, cuando un CIM es empleado de manera racional por sus usuarios, la mayoría de sus consultas deberían relacionarse con la Atención Farmacéutica. Objetivo: Establecer si las consultas efectuadas al CIM de Bizkaia están relacionadas con la Atención Farmacéutica. Métodos: Se ha elaborado una tabla que relaciona la temática de las consultas con la AF y se han analizado con esta herramienta las consultas efectuadas en 2001, procesando los datos con Microsoft Excel®. Resultados: De las 10.929 consultas recibidas, 8.866 (81,12%) estaban relacionadas con la AF, en 558 (5,11%) no pudo determinarse si existía relación, debido a la metodología del estudio empleada, y 1.505 (13,77%) versaron sobre otros temas, las más numerosas sobre nutrición y dietética (16,50%). Conclusión: Dada la coincidencia de los objetivos de un CIM con los de la Atención Farmacéutica y la relación existente entre las consultas que se reciben en el CIM de Bizkaia y la Atención Farmacéutica, puede deducirse que un CIM puede ser un recurso importante para el desarrollo de esta modalidad de práctica en la farmacia (AU)

Introduction: The goal of a Drug Information Center (DIC) is to supply information intended to foster a rational use of drugs. Its main activity is to solve queries about drugs. The users of the Pharmacists’ Asociations DICs are basically community pharmacists. The latter should focus their daily practice towards the pharmaceutical care, that is, dispensing, pharmaceutical consultation, pharmacotherapy follow-up… All of these activities are patient focused and closely related to the rational use of drugs. Therefore, the goal of the DICs and those of the fomer activities come together in a great extent. Hence, when DIC users make a rational use of this service, most of their information queries should be related to pharmaceutical care issues. Objective: To establish if the drug information queries addressed to the Biscaian DIC are related to pharmaceutical care. Methods: A table was built where the subject of the queries were linked with pharmaceutical care. This instrument was used to evaluate all the queries addressed to the DIC throughout the 2001 year and data were analized with Microsoft Excel®. Results: During this year, the Biscaian DIC received 10929 drug information queries. Overall, 8866 (81,12%) were related to pharmaceutical care and additionally there was not posible to establish a direct relationship with pharmaceutical care in another 558 queries (5,11%) due to problems with the methodology used.1505 (13,77%) queries were related to other issues, mainly about nutrition and dietetics (16,50% of the latter). Conclusion: As the objectives of a DIC collate with those of pharmaceutical care, along with the fact that the queries received in the Biscaian DIC are related with pharmaceutical care, we can conclude that a DIC can be a core resource for the development of this kind of pharmacy practice (AU)

Coronary artery disease diagnosis based on exercise electrocardiogram indexes from repolarisation, depolarisation and heart rate variability.

Several indexes have been reported to improve the accuracy of exercise test electrocardiogram (ECG) analysis in the diagnosis of coronary artery disease (CAD), compared with the classical ST depression criterion. Some of them combine repolarisation measurements with heart rate (HR) information (such as the so-called ST/HR hysteresis); others are obtained from the depolarisation period (such as the Athens QRS score); finally, there are heart rate variability (HRV) indexes that account for the nervous system activity. The aim of this study was to identify the best exercise ECG indexes for CAD diagnosis. First, a method to automatically estimate repolarisation and depolarisation indexes in the presence of noise during a stress test was developed. The method is divided into three stages: first, a preprocessing step, where QRS detection, filtering and baseline beat rejection are applied to the raw ECG, prior to a weighted averaging; secondly, a post-processing step in which potentially noisy averaged beats are identified and discarded based on their noise variance; finally, the measurement step, in which ECG indexes are computed from the averaged beats. Then, a multivariate discriminant analysis was applied to classify patients referred for the exercise test into two groups: ischaemic (positive coronary angiography) and low-risk (Framingham risk index < 5%). HR-corrected repolarisation indexes improved the sensitivity (SE) and specificity (SP) of the classical exercise test (SE = 90%, SP = 79% against SE = 65%, SP = 66%). Depolarisation indexes also achieved an improvement over ST depression measurements (SE = 78%, SP = 81%). HRV indexes obtained the best classification results in our study population (SE = 94%, SP = 92%) by means of the very high-frequency power (VHF) (0.4-1 Hz) at stress peak.

Precise mapping of a locus affecting grain protein content in durum wheat.

Grain protein content (GPC) is an important factor in pasta and breadmaking quality, and in human nutrition. It is also an important trait for wheat growers because premium prices are frequently paid for wheat with high GPC. A promising source for alleles to increase GPC was detected on chromosome 6B of Triticum turgidum var. dicoccoides accession FA-15-3 (DIC). Two previous quantitative trait locus (QTL) studies found that the positive effect of DIC-6B was associated to a single locus located between the centromere and the Nor-B2 locus on the short arm of chromosome 6B. Microsatellite markers Xgwm508 and Xgwm193 flanking the QTL region were used in this study to develop 20 new homozygous recombinant substitution lines (RSLs) with crossovers between these markers. These 20 RSLs, plus nine RSLs developed in previous studies were characterized with four new RFLP markers located within this chromosome segment. Grain protein content was determined in three field experiments organized as randomized complete block designs with ten replications each. The QTL peaks for protein content were located in the central region of a 2.7-cM interval between RFLP markers Xcdo365 and Xucw67 in the three experiments. Statistical analyses showed that almost all lines could be classified unequivocally within low- and high- protein groups, facilitating the mapping of this trait as a single Mendelian locus designated Gpc-6B1. The Gpc-6B1 locus was mapped 1.5-cM proximal to Xcdo365 and 1.2-cM distal to Xucw67. These new markers can be used to reduce the size of the DIC chromosome segment selected in marker-assisted selection programs. Markers Nor-B2 and Xucw66 flanking the previous two markers can be used to select against the DIC segment and reduce the linkage drag during the transfer of Gpc-6B1 into commercial bread and pasta wheat varieties. The precise mapping of the high GPC gene, the high frequency of recombinants recovered in the targeted region, and the recent development of a tetraploid BAC library including the Gpc-6B1 DIC allele are the first steps towards the map-based cloning of this gene.

Compartmentalisation between gut and lung mucosae in a model of secondary immunodeficiency: effect of thymomodulin.

UNLABELLED: Compartmentalisation of mucosal immune response seems to be the result mainly of the preferential migration of activated cells back to their inductive sites. The aim of this report was to demonstrate, in a model of secondary immunodeficiency in Wistar rats (severely protein deprived at weaning and refed with casein 20%; group R21), that the oral administration of Thymomodulin (group:R21TmB) has different effects on gut and BALT (Bronchus-associated lymphoid tissue). Tissue sections (5 mu) were studied by immunohistochemistry 1). The oral administration of Thymomodulin restores only in gut Lamina propria (LP) the IgA B and CD4 T cell populations to control levels. The CD8a and CD25 subpopulations do not vary in gut as they return to control levels when refed with 20% casein diet. All the populations mentioned above remained decreased even after receiving Thymomodulin by the oral route. However, the same behaviour was observed for the TCR delta T cells that were decreased and return to normal levels in both mucosae by the effect of the immunomodulator; 2) when studying the iIEL (intestinal intraepithelial lymphocytes) CD8 alpha, CD25 and TCR gamma delta T cells, that were increased in R21, return to control levels in R21TmB. In BALT intraepithelium CD8 alpha and CD25 T cells remained decreased, while only TCR gamma delta T cells (increased in R21) return to control values. CONCLUSIONS: 1) there exists a compartmentalisation between both mucosae, as T CD4+ and IgA B+ cells are restored by TmB only in gut; 2) only those iIEL involved in inflammation (CD8 alpha+/CD25+ and TCR gamma delta+/CD25+) are normalised by means of the Thymomodulin 3) however, in BALT,only TCR gamma delta+ T cells are restored 4) the oral administration of the present immunomodulator may be useful as a therapeutic agent, although the preferential survival in the tissue of initial stimulation is the major factor in the preferential distribution of activated cells.

Effect of retinal ablation on the expression of calbindin D28k and GABA in the chick optic tectum.

The effect of retinal ablation on qualitative and quantitative changes of calbindin D28k and GABA expression in the contralateral optic tectum was studied in young chicks. Fifteen days old chicks had unilateral retinal ablation and after 7 or 15 days, calbindin expression was analyzed by Western blot and immunocytochemistry. Neuronal degeneration was followed by the amino-cupric silver technique. After 15 days, retinal lesions produced a significant decrease in calbindin immunostaining in the neuropil of layers 5-6 and in the somata of neurons from the layers 8 and 10 of the contralateral tectum, being this effect less marked at 7 days post-lesion. Double staining revealed that 50-60% of cells in the layers 8 and 10 were calbindin and GABA positive, 30-45% were only calbindin positive and 5-10% were only GABAergic neurons. Retinal ablation also produced a decrease in the GABA expression at either 7 or 15 days after surgery. At 7 days, dense silver staining was observed in the layers 5-6 from the optic tectum contralateral to the retinal ablation, which mainly represented neuropil that would come from processes of retinal ganglion cells. Tectal neuronal bodies were not stained with silver, although some neurons were surrounded by coarse granular silver deposits. In conclusion, most of calbindin molecules are present in neurons of the tectal GABAergic inhibitory circuitry, whose functioning apparently depends on the integrity of the visual input. A possible role of calbindin in the control of intracellular Ca2+ in neurons of this circuit when the visual transmission arrives to the optic tectum remains to be studied.

Cytogenetic studies in bone marrow cells from Wistar rats in protein malnutrition.

The effect of severe protein deficiency at weaning has been studied in bone marrow, which is a primary lymphoid organ. Our experimental model of secondary immunodeficiency in Wistar rats has shown: (1) a decreased number of viable bone marrow cells (P <.0001); (2) diminished percentage of mitosis (P <.01); and (3) severe alteration in the percentage of chromosome pairs 3, 11, and 12 bearing nucleolar organizer regions (NORs) (P <.05). This last finding indicates a poor ribosomal gene activity. These alterations were reverted after the oral administration of a 20% casein diet during 5 to 9 days. However, there were no karyotype variations between the experimental groups. We conclude from these results that severe protein deficiency at weaning alters several aspects of bone marrow cell proliferation and ribosomal gene activity as determined by the number of silver stained nucleolus organizer regions.

Metastasis to the breast from extramammary malignancies: a report of four cases and a review of literature.

Breast metastases from extramammary tumours are rare with few cases reported. Four cases of metastasis to the breast are presented and the diagnostic problems of this condition are reviewed. Correlation between the histology of primary tumour and the cytology of breast metastatic tumour can avoid the surgical breast biopsy and unnecessary mastectomy. Metastasis to the breast has poor prognosis.

Flow cytometric analysis of intestinal intraepithelial lymphocytes in a model of immunodeficiency in Wistar rats.

BACKGROUND: We have shown, in a rat model of immunodeficiency, permanent alterations in the thymus and in the gut-associated lymphoid tissues. We observed by immunohistochemistry an increase in the number of gamma/delta+ T cells in the gut lamina propria and in the number of CD8alpha/alpha+, CD25+, gamma/delta+ subpopulations of intestinal intraepithelial lymphocytes (iIEL). The aim of the present study was to analyze the isolated rat iIEL by flow cytometry. Materials and Methods Cells from mesenteric lymph nodes were examined in parallel with isolated iIEL. After staining with different antibodies, samples were run on a FACScan flow cytometer. Background staining was evaluated using isotype controls. Data analysis was performed using Lysys II software (Becton Dickinson) and WinMDI 2.3 software. RESULTS: 1) CD8alpha/beta populations do not express TCRgamma/delta, 2) CD8alpha/alpha+ populations express TCRgamma/delta, and its percentage is significantly increased in R21, 3) CD8alpha/beta and CD8alpha/alpha iIEL express TCRalpha/beta, being the percentage of CD8alpha/alpha+ TCRalpha/beta+ iIEL increased and the percentage of CD8alpha/beta+ TCRalpha/beta+ iIEL decreased in R21, and 4) CD8alpha/alpha as well as CD8alpha/beta iIEL do express CD25 only in R21. CONCLUSIONS: Considering the above results, we conclude that there exists an "in situ" origin and extrathymic maturation of the CD8alpha/alpha+ iIEL in the intestinal epithelium. The increase of TCRgamma/delta+ T cells may be triggered by the carbohydrate dextrin, to provide immune protection and control of inflammation at the intestinal level.