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Rationale: Cigarette smoke is considered the chief leading cause of chronic obstructive pulmonary disease (COPD). Its impact on the progressive deterioration of airways has been extensively studied...

Staphylococcus aureus is the most common cause of surgical site infections and its treatment is challenging due to the emergence of multi-drug resistant strains such as methicillin-resistant S. aureus (MRSA). Natural berry-derived compounds have shown antimicrobial potential, e.g., ellagitannins such as sanguiin H-6 and lambertianin C, the main phenolic compounds in Rubus seeds, have shown antimicrobial activity. The aim of this study was to evaluate the effect of sanguiin H-6 and lambertianin C fractionated from cloudberry seeds, on the MRSA growth, and as treatment of a MRSA biofilm development in different growth media in vitro and in vivo by using a murine wound infection model where sanguiin H-6 and lambertianin C were used to prevent the MRSA infection. Sanguiin H-6 and lambertianin C inhibited the in vitro biofilm development and growth of MRSA. Furthermore, sanguiin H-6 showed significant anti-MRSA effect in the in vivo wound model. Our study shows the possible use of sanguiin H-6 as a preventive measure in surgical sites to avoid postoperative infections, whilst lambertianin C showed no anti-MRSA activity.

Smoking is considered the chief leading cause of chronic obstructive pulmonary disease (COPD). Its impact on the progressive deterioration of the airways has been extensively studied. However, its direct effects on pulmonary vasculature have not been studied in deep, in part due to the classic conception that vascular damage is a consequence of alveolar hypoxia and loss of capillary bed. We have used different molecular and cell biology approaches, as well as tissue studies from in vivo models and human samples to analyse how cigarette smoke extract (CSE) regulated pulmonary arterial cells phenotypic modulation, particularly on human fibroblasts and smooth muscle cells. Our results demonstrate that CSE exposure has direct effects on hPAFib and hPASMC, promoting a senescent phenotype that in turn contributes to increase the proliferative potential of non-exposed cells. Using traction force microscopy, wire myography and patch-clamp techniques, we demonstrate that CSE affects cell contractility and dysregulates the expression and activity of the voltage-gated K+ channel Kv7.4. This ultimately contributes to limit vascular responses impairing vasoconstriction and endothelium-dependent and independent relaxation. Additionally, the levels of this channel are diminished in the lungs of smoke-exposed mice and COPD patients. These findings greatly enhance knowledge about pulmonary diseases, in particular COPD, and open a new line of study related to the exploitation of these mechanisms as therapeutic targets.

Introduction Within the pathogenesis of the chronic obstructive pulmonary disease (COPD) there are interactions between different inflammatory mediators that are enhanced during an exacerbation. Arginase is present in bronchial epithelial cells, endothelial, fibroblasts and alveolar macrophages, which make it a probable key enzyme in the regulation of inflammation and remodelling. We aimed to find a potential relationship between arginase activity, inflammatory mediators in COPD patients in stable phase and during exacerbations. Methods We performed a prospective, observational study of cases and controls, with 4 study groups (healthy controls, stable COPD, COPD during an exacerbation and COPD 3 months after exacerbation). We measured arginase, inflammation markers (IL-6, IL-8, TNF-∝, IFN-γ and C reactive protein), and mediators of immunity: neutrophils, monocytes, total TCD3 + lymphocytes (CD3ζ), CD4 + T cells, CD8 + T cells, NK cells. Results A total of 49 subjects were recruited, average age of 69.73 years (59.18% male). Arginase activity is elevated during an exacerbation of COPD, and this rise is related to an increase in IL-6 production. The levels of IL-6 and IL-8 remained elevated in patients with COPD at 3 months after hospital exacerbation. We did not find a clear relationship between arginase activity, immunity or with the degree of obstruction in COPD patients. Conclusions Arginase activity is elevated during an exacerbation of COPD, and it could be related to an increase in the production of IL-6. Levels of IL-6, IL-8, and arginase activity remain elevated in patients with COPD at 3 months after hospital exacerbation. Arginase activity could contribute to the development of COPD.

Skeletal muscle dysfunction is a common complication and an important prognostic factor in patients with chronic obstructive pulmonary disease (COPD). It is associated with intrinsic muscular abnormalities of the lower extremities, but it is not known whether there is an easy way to predict its presence. Using a mouse model of chronic cigarette smoke exposure, we tested the hypothesis that magnetic resonance spectroscopy allows us to detect muscle bioenergetic deficit in early stages of lung disease. We employed this technique to evaluate the synthesis rate of adenosine triphosphate (ATP) and characterize concomitant mitochondrial dynamics patterns in the gastrocnemius muscle of emphysematous mice. The fibers type composition and citrate synthase (CtS) and cytochrome c oxidase subunit IV (COX4) enzymatic activities were evaluated. We found that the rate of ATP synthesis was reduced in the distal skeletal muscle of mice exposed to cigarette smoke. Emphysematous mice showed a significant reduction in body weight gain, in the cross-sectional area of the total fiber and in the COX4 to CtS activity ratio, due to a significant increase in CtS activity of the gastrocnemius muscle. Taken together, these data support the hypothesis that in the early stage of lung disease, we can detect a decrease in ATP synthesis in skeletal muscle, partly caused by high oxidative mitochondrial enzyme activity. These findings may be relevant to predict the presence of skeletal bioenergetic deficit in the early stage of lung disease besides placing the mitochondria as a potential therapeutic target for the treatment of COPD comorbidities.

Skeletal muscle dysfunction is a common feature in COPD which is associated with intrinsic muscular abnormalities, but it is not known whether there is an easy way of predicting their presence in early stage of disease. Using a mouse model of cigarette smoke exposure, we tested the hypothesis that 31-phosphorus magnetic resonance spectroscopy allows us to early detect skeletal muscle bioenergetic deficit. We employed this technique to assess the rate of ATP synthesis, and characterized the concomitant mitochondrial dynamics patterns in skeletal. We specifically explored whether experimental COPD model may alter mitochondrial respiratory chain complexes in peripheral muscles. We evaluated citrate synthase (CtS) and cytochrome c oxidase (COX) enzyme activities in gastrocnemius of mice. We found that rate of muscle ATP synthesis was reduced in skeletal muscle of mice exposed to cigarette smoke. Furthermore, ATP synthesis rate was directly related to mitochondrial enzyme activity. Emphysematous mice showed a significant reduction in body weight gain and decreased activity of CtS/COX. Taken together, these data support the hypothesis that in early stage of lung disease we are able to detect a decreased skeletal muscle ATP synthesis, in part caused by low mitochondrial enzyme activity. These findings may prove relevant to predict the skeletal bioenergetic deficit presence in the early stage of the lung disease and thus be able to act preventively in the evolution of the disease, besides placing mitochondria as potential therapeutic target for the treatment of COPD comorbidities.

The bile acid conjugate tauroursodeoxycholic acid (TUDCA) is a neuroprotective agent in various animal models of neuropathologies. We have previously shown the anti-inflammatory properties of TUDCA in an animal model of acute neuroinflammation. Here, we present a new anti-inflammatory mechanism of TUDCA through the regulation of transforming growth factor β (TGFβ) pathway. The bacterial lipopolysaccharide (LPS) was injected intravenously (iv) on TGFβ reporter mice (Smad-binding element (SBE)/Tk-Luc) to study in their brains the real-time activation profile of the TGFβ pathway in a non-invasive way. The activation of the TGFβ pathway in the brain of SBE/Tk-Luc mice increased 24 h after LPS injection, compared to control animals. This activation peak increased further in mice treated with both LPS and TUDCA than in mice treated with LPS only. The enhanced TGFβ activation in mice treated with LPS and TUDCA correlated with both an increase in TGFβ3 transcript in mouse brain and an increase in TGFβ3 immunoreactivity in microglia/macrophages, endothelial cells, and neurons. Inhibition of the TGFβ receptor with SB431542 drug reverted the effect of TUDCA on microglia/macrophages activation and on TGFβ3 immunoreactivity. Under inflammatory conditions, treatment with TUDCA enhanced further the activation of TGFβ pathway in mouse brain and increased the expression of TGFβ3. Therefore, the induction of TGFβ3 by TUDCA might act as a positive feedback, increasing the initial activation of the TGFβ pathway by the inflammatory stimulus. Our findings provide proof-of-concept that TGFβ contributes to the anti-inflammatory effect of TUDCA under neuroinflammatory conditions.

This study aims to develop an NMR-based metabonomic approach to diagnostic evaluation of Chronic Obstructive Pulmonary Disease (COPD) in a model of smoke exposure in mice. The current study compared the effects of acute and chronic smoke exposures performed on three strains of mice susceptible to develop COPD and two strains resistant to this disease. Intact lung tissue was examined using high-resolution magic angle spinning (HR-MAS). Six chemical shifts of the 1H NMR spectrum including taurine, glutathione, phosphoryl choline and glycero-phosphocholine, were selected to ensure a high discrimination between control and acute exposed mice in the susceptible strain and, simultaneously, a low discrimination in the resistant strains. Using these metabolites, a chronic predictive model was developed and validated. The multivariate analysis provides a 100% true classification for sick mice and 78% for control animals. This pioneering study demonstrates that the NMR-based metabonomic analysis of intact lung tissue is a potential technique for the detection of COPD biomarkers and for the diagnosis of COPD in a mouse model of COPD. It might boost the study of tobacco smoke effects on other non- or less-invasive samples.

Resumen Introduccion El liver growth factor (LGF, ‘factor de crecimiento de higado’) es un mitogeno con actividad regeneradora y antifibrotica con actividad incluso en localizaciones extrahepaticas. En este trabajo se administro LGF en un modelo de fibrosis pulmonar inducido con cloruro de cadmio (CdCl2) para estudiar su capacidad antifibrotica. Metodos Se instilaron 42 ratas Wistar macho con 0,5 ml/rata de CdCl2 al 0,025% (n=21) o de salino (n=21). Transcurridos 35 dias y una vez establecida la lesion se realizo el tratamiento con LGF y posteriormente el analisis de los parametros funcionales —capacidad inspiratoria (CI), complianza pulmonar (CL), capacidad vital forzada (CVF) y flujo espiratorio forzado al 75% (FEF75%)—, morfometria —area interna alveolar y distancia media entre paredes alveolares— y contenido en colageno y elastina. Resultados La fibrosis pulmonar originada mediante CdCl2 se caracterizo por un marcado descenso de la funcion pulmonar en comparacion con los controles. Se redujo un 28% la CI, un 38% la CL, un 31% la CVF y un 54% la FEF75%, descenso que se revirtio parcialmente tras la inyeccion de LGF —el 18% en CI, el 27% en CL, el 19% en CVF y el 35% en FEF75%—. Ademas, se observo un incremento en la cantidad de colageno y elastina del 165 y el 76%, respectivamente, en las ratas del grupo CdCl2 frente a un 110 y un 34% tras la inyeccion de LGF. Conclusiones Estos datos demuestran que el LGF mejora la funcion pulmonar y revierte parcialmente el incremento de las proteinas de matriz pulmonar producido por la instilacion con CdCl2.

Inflammation is one of the first immune system responses to any type of aggression. As with any type of aggression, the lesion produced by inhalation of tobacco smoke prompts an innate inflammatory response. Subsequently, this lesion is stimulated by the release of various chemical factors that enhance the inflammatory response and, finally--depending on the type of aggression--acquired immunity is activated, which, mediated by lymphocyte participation, serves to establish a physical barrier against the propagation of the lesion and to aid repair of the damaged pulmonary tissue. However, the balance between inflammation and repair is not always maintained, as is the case in chronic obstructive pulmonary disease (COPD), in which marked changes appear in the architecture of the airways, alveolar spaces and pulmonary arteries, forming the structural background of the functional changes characteristic of this disease. COPD is basically a pulmonary disease but data are available on the existence of associated systemic inflammation. The origins of this systemic inflammation are unclear: some information indicates that tobacco smoke is a direct origin common to local and systemic inflammation, while other data point to primary pulmonary inflammation that secondarily produces systemic involvement. The present review describes the main mechanisms involved in both pulmonary and systemic inflammation in COPD.

The aim of this study was to examine the differential regulation after acute ethanol administration on tyrosine hydroxylase, proenkephalin and cannabinoid CB1 receptor gene expressions in selected areas of the rat brain. Rats received an intragastric administration of 3 g/kg ethanol and were killed by decapitation at 1, 2, 4, 8 and 24 h. The results showed an activation of tyrosine hydroxylase gene expression in the ventral tegmental area and the substantia nigra, increased proenkephalin gene expression in the caudate-putamen, nucleus accumbens core and shell, central and medial amygdala, ventromedial hypothalamic nucleus and the paraventricular hypothalamic nucleus. In contrast, a significant decrease in the cannabinoid CB1 receptor gene expression was found in caudate-putamen, central amygdala and ventromedial hypothalamic nucleus. In conclusion, the results suggest that an acute dose of ethanol induces neuroplastic alterations in proenkephalin, tyrosine hydroxylase and cannabinoid CB1 receptor gene expressions that may contribute to trigger the rewarding effects of ethanol consumption. © 2007 Elsevier B.V. and ECNP., This research was supported by grants from the “Plan Nacional Sobre Drogas” (National Plan on Drugs) from the Ministerio del Interior and FIS 01/1438, FIS 03/216 and FIS 05/429 from the Spanish Ministry of Health to J. Manzanares.

Over the past few years, advances in the investigation of the neurochemical circuits involved in the development and treatment of alcohol dependence have identified peptides and receptors as potential key targets in the treatment of problems related to alcohol consumption. The endogenous opioid system is modified by alcohol intake in areas of the brain related to reward systems, and differential basal levels of opioid gene expression are found in rodents with a high preference for ethanol. This suggests a greater vulnerability to alcohol consumption in relation to differences in genetic background. Further evidence of the involvement of opioid peptides in alcohol dependence is the ability of the opioid antagonist naltrexone to reduce alcohol intake in animal models of dependence and in alcohol-dependent patients. Abundant evidence indicates that the activation of cannabinoid receptors stimulates the release of opioid peptides, therefore the cannabinoid receptor antagonists may presumably alter opioid peptide release, thus facilitating the reduction of ethanol consumption. However, little is known about the effects of ethanol on the endogenous cannabinoid system, the vulnerability of cannabinoid receptors to alcohol intake or their neurochemical implications in reducing consumption of alcohol. In this paper, we review the role of opioid and cannabinoid receptor systems, their vulnerability to alcohol intake and the development of dependence, and the targeting of these systems in the treatment of alcoholism.

Aim: To specify the functional activity of cannabinoid CB 1 receptor in alcohol-preferring Fawn Hooded and alcohol nonpreferring Wistar rats under naive conditions. Method: Cannabinoid CB1 (WIN-55,212)-stimulated ( 35 S)-GTPs binding autoradiography, and cannabinoid CB 1 receptor gene expression were measured in rats of both strains that received only water. Results: Cannabinoid CB1 receptor stimulated ( 35 S)- GTPs binding was significantly lower in cingulate cortex (Cg), caudate-putamen (CPu), nucleus accumbens (Acc), ventromedial hypothalamic nucleus (VMN), amygdaloid area (AMG), fields (CA1, CA3) of the hippocampus and dentate gyrus (DG) in Fawn Hooded than in Wistar rats, whereas no differences were found either in substantia n igra pars reticulata (SNr) nor CA2 field of the hippocampus. In addition, cannabinoid CB 1 receptor gene expression was lower in Cg, CPu, VMN and CA3 field of the hippocampus in Fawn Hooded than in Wistar rats. Conclusions: We speculate that lower cannabinoid function appears to be related to greater vulnerability to alcohol consumption. Cannabinoid CB 1 receptor may represent a key target in the treatment of alcohol dependence.

Resumen Objetivo El proceso de dependencia al alcohol implica modificaciones funcionales en la neurotransmision cerebral alterando de esta manera el equilibrio homeostatico entre neurotransmisores inhibidores y excitadores en diferentes circuitos cerebrales relacionados con el refuerzo inducido por sustancias. Material y metodos Se revisa la neurobiologia de la dependencia al alcohol. Resultados Un gran numero de resultados experimentales sugieren que el alcohol ejerce efectos muy variados sobre los principales sistemas de neurotransmision dependiendo del tiempo y el patron de ingesta de la bebida. Asi, el consumo de alcohol a corto plazo potencia la neurotransmision inhibitoria (por inhibicion de la transmision glutamatergica, y activacion de neuronas GABAergicas), mientras que a largo plazo se produce tolerancia, lo que tiende a desplazar estas alteraciones neuroquimicas hacia valores normales. Por el contrario, si el consumo de alcohol es discontinuo o se reduce bruscamente, se produce activacion de la neurotransmision glutamatergica e inhibicion de la neurotransmision GABAergica, entre otras muchas alteraciones neuroquimicas, lo que desencadena sindrome de abstinencia y consiguiente comportamiento de busqueda y deseo irrefrenable de consumir alcohol. Conclusiones Los efectos del alcohol estan regulados por distintos sistemas de neurotransmision y neuromodulacion, entre los que se puede destacar los sistemas de la dopamina (DA), noradrenalina (NA), factor liberador de corticotropina (CRF), neuropeptido Y (NPY), sistema opioide y el sistema cannabinoide endogeno, de forma que no se pueden explicar muchos de los efectos farmacologicos y comportamentales del alcohol sin entender sus acciones sobre estos sistemas.

Chronic obstructive pulmonary disease (COPD) is an inflammatory lung disease largely associated with cigarette smoke exposure (CSE) and characterized by pulmonary and extrapulmonary manifestations, including systemic inflammation. Liver growth factor (LGF) is an albumin-bilirubin complex with demonstrated antifibrotic, antioxidant, and antihypertensive actions even at extrahepatic sites. We aimed to determine whether short LGF treatment (1.7 μg/mouse ip; 2 times, 2 wk), once the lung damage was established through the chronic CSE, contributes to improvement of the regeneration of damaged lung tissue, reducing systemic inflammation. We studied AKR/J mice, divided into three groups: control (air-exposed), CSE (chronic CSE), and CSE + LGF (LGF-treated CSE mice). We assessed pulmonary function, morphometric data, and levels of various systemic inflammatory markers to test the LGF regenerative capacity in this system. Our results revealed that the lungs of the CSE animals showed pulmonary emphysema and inflammation, characterized by increased lung compliance, enlargement of alveolar airspaces, systemic inflammation (circulating leukocytes and serum TNF-α level), and in vivo lung matrix metalloproteinase activity. LGF treatment was able to reverse all these parameters, decreasing total cell count in bronchoalveolar lavage fluid and T-lymphocyte infiltration in peripheral blood observed in emphysematous mice and reversing the decrease in monocytes observed in chronic CSE mice, and tends to reduce the neutrophil population and serum TNF-α level. In conclusion, LGF treatment normalizes the physiological and morphological parameters and levels of various systemic inflammatory biomarkers in a chronic CSE AKR/J model, which may have important pathophysiological and therapeutic implications for subjects with stable COPD.

Animal models of disease have always been welcomed by the scientific community because they provide an approach to the investigation of certain aspects of the disease in question. Animal models of COPD cannot reproduce the heterogeneity of the disease and usually only manage to represent the disease in its milder stages. Moreover, airflow obstruction, the variable that determines patient diagnosis, not always taken into account in the models. For this reason, models have focused on the development of emphysema, easily detectable by lung morphometry, and have disregarded other components of the disease, such as airway injury or associated vascular changes. Continuous, long-term exposure to cigarette smoke is considered the main risk factor for this disease, justifying the fact that the cigarette smoke exposure model is the most widely used. Some variations on this basic model, related to exposure time, the association of other inducers or inhibitors, exacerbations or the use of transgenic animals to facilitate the identification of pathogenic pathways have been developed. Some variations or heterogeneity of this disease, then, can be reproduced and models can be designed for resolving researchers' questions on disease identification or treatment responses.

This study investigates the role of proinflammatory monocytes recruited from blood circulation and recovered in bronchoalveolar lavage (BAL) fluid in mediating the lung damage in a model of acute cigarette smoke (CS)-induced lung inflammation in two strains of mice with different susceptibility to develop emphysema (susceptible -C57BL/6J and non susceptible -129S2/SvHsd). Exposure to whole-body CS for 3 consecutive research cigarettes in one single day induced acute inflammation in the lung of mice. Analysis of BAL fluid showed more influx of recently migrated monocytes at 72 h after CS-exposition in susceptible compared to non susceptible mice. It correlated with an increase in MMP-12 and TNF-α protein levels in the lung tissue, and with an increment of NF-κB translocation to the nucleus measured by electrophoretic mobility shift assay in C57BL/6J mice. To determine the functional role of these proinflammatory monocytes in mediating CS-induced airway inflammation, alveolar macrophages and blood monocytes were transiently removed by pretreatment with intratracheal and intravenous liposome-encapsulated CL2MDP, given 2 and 4 days prior to CS exposure and their repopulation was studied. Monocytes/macrophages were maximally depleted 48 h after last liposome application and subsequently recently migrated monocytes reappeared in BAL fluid of susceptible mice at 72 h after CS exposure. Recently migrated monocytes influx to the lung correlated with an increase in the MMP-12 protein level in the lung tissue, indicating that the increase in proinflammatory monocytes is associated with a major tissue damaging. Therefore our data confirm that the recruitment of proinflammatory recently migrated monocytes from the blood are responsible for the increase in MMP-12 and has an important role in the pathogenesis of lung disease induced by acute lung inflammation. These results could contribute to understanding the different susceptibility to CS of these strains of mice, The work presented was supported by the Spanish ‘‘Ministry of Science and Innovation’’ (SAF2008-05412-C02) and ‘‘Spanish Society of Pneumology and Thoracic Surgery’’ (SEPAR-1002)

Cigarette smoke (CS)-induced oxidative stress may cause muscle alterations in chronic conditions such as chronic obstructive pulmonary disease (COPD). We sought to explore in AKR/J mice exposed to CS for 6 months and in control animals, levels of protein oxidation, oxidized proteins (immunoblotting, proteomics) and antioxidant mechanisms in both respiratory and limb muscles, body weight modifications, systemic inflammation, and lung structure. Compared to control mice, CS-exposed animals exhibited a reduction in body weight gain at 3 months and thereafter, showed lung emphysema, and exhibited increased oxidative stress levels in their diaphragms and gastrocnemius at 6 months. Proteins involved in glycolysis, ATP production and distribution, carbon dioxide hydration, and muscle contraction were carbonylated in respiratory and limb muscles. Blood tumor necrosis factor (TNF)-alpha levels were significantly greater in CS-exposed mice than in control animals. In AKR/J mice, chronic exposure to CS induces lung emphysema concomitantly with greater oxidative modifications on muscle proteins in both respiratory and limb muscles, and systemic inflammation.

Inflammation is one of the first immune system responses to any type of aggression. As with any type of aggression, the lesion produced by inhalation of tobacco smoke prompts an innate inflammatory response. Subsequently, this lesion is stimulated by the release of various chemical factors that enhance the inflammatory response and, finally--depending on the type of aggression--acquired immunity is activated, which, mediated by lymphocyte participation, serves to establish a physical barrier against the propagation of the lesion and to aid repair of the damaged pulmonary tissue. However, the balance between inflammation and repair is not always maintained, as is the case in chronic obstructive pulmonary disease (COPD), in which marked changes appear in the architecture of the airways, alveolar spaces and pulmonary arteries, forming the structural background of the functional changes characteristic of this disease. COPD is basically a pulmonary disease but data are available on the existence of associated systemic inflammation. The origins of this systemic inflammation are unclear: some information indicates that tobacco smoke is a direct origin common to local and systemic inflammation, while other data point to primary pulmonary inflammation that secondarily produces systemic involvement. The present review describes the main mechanisms involved in both pulmonary and systemic inflammation in COPD.

This study evaluated the role of prodynorphin gene in the regulation of anxiety and associated molecular mechanisms. Emotional responses were assessed using the light-dark test, elevated plus maze and social interaction tests in prodynorphin knockout and wild-type mice. Corticotrophin releasing factor and proopiomelanocortin gene expressions in the hypothalamus were evaluated after restraint stress using in situ hybridization. The anxiolytic efficacy of bromazepam and GABA A receptor subunits gene expression in the amygdala were also assessed in both genotypes. The deletion of prodynorphin increased anxiety-like behaviours and proopiomelanocortin gene expression in the arcuate nucleus (two-fold). Moreover, the anxiolytic action of bromazepam was significantly attenuated in the mutant mice. Decreased GABA Aγ 2 and increased GABA Aβ 2 gene expression receptor subunits were found in the amygdala of prodynorphin knockout mice. These results indicate that deletion of prodynorphin gene is associated with increased anxiety-like behaviours, enhanced sensibility response to stress stimuli, reduced anxiolytic efficacy of bromazepam and altered expression of the GABA A receptor subunits.

Introduction Liver growth factor (LGF) is a liver mitogen with regenerating and anti-fibrotic activity even at extrahepatic sites. We used LGF in a lung fibrosis model induced by cadmium chloride (CdCl2), to study its antifibrotic capacity. Methods Forty-two male Wistar rats were administered a single dose of 0.5ml/rat of CdCl2 0.025% (n = 21) or the same volume of saline (control group, n = 21). After 35 days, once a lesion was established, we started a 3 week treatment with LGF, after which we determined lung function ––inspiratory capacity (IC), lung compliance (LC), forced vital capacity (FVC) and expiratory flow at 75% (FEF75%)––, lung morphometry ––alveolar internal area (AIA), mean linear intersection (LM)––, and collagen (both by Sirius red and hydroxyproline residues) and elastin contents. Results Pulmonary fibrosis in CdCl2 rats was characterized by a marked decrease in pulmonary function with respect to healthy controls ––reductions of 28% in IC, 38% in CL, 31% in FVC, and 54% in FEF75%–– which was partially recovered after LGF injection ––18% IC, 27% CL, 19% FVC and 35% FEF75%––; increase in collagen and elastin contents ––165% and 76%, respectively, in CdCl2 rats, versus 110% and 34% after LGF injection––; and increases in AIA and LM, partially inverted by LGF. Conclusions Together, these data seem to demonstrate that LGF is able to improve lung function and partially inverts the increase in lung matrix proteins produced by CdCl2 instillation.

Motor impairment, dopamine (DA) neuronal activity and proenkephalin (PENK) gene expression in the caudate-putamen (CPu) were measured in 6-OHDA-lesioned and treated (l-DOPA. +. benserazide) CB1 KO and WT mice. A lesion induced by 6-OHDA produced more severe motor deterioration in CB1 KO mice accompanied by more loss of DA neurons and increased PENK gene expression in the CPu. Oxidative/nitrosative and neuroinflammatory parameters were estimated in the CPu and cingulate cortex (Cg). CB1 KO mice exhibited higher MDA levels and iNOS protein expression in the CPu and Cg compared to WT mice. Treatment with l-DOPA. +. benserazide (12 weeks) resulted in less severe dyskinesias in CB1 KO than in WT mice.The results revealed that the lack of cannabinoid CB1 receptors increased the severity of motor impairment and DA lesion, and reduced l-DOPA-induced dyskinesias. These results suggest that activation of CB1 receptors offers neuroprotection against dopaminergic lesion and the development of l-DOPA-induced dyskinesias. © 2009 Elsevier Inc., This research was supported by grants from “FUNDACIÓN la CAIXA” and “Fundación de Neurociencias y Envejecimiento” to JM, the “CATEDRADISTEC” for research in Parkinson’s disease (JM and CL) and to CAM S-SAL/0261/2006 (JCL).SPR was a predoctoral fellow supported by “Fundación la CAIXA”. MSGG is a predoctoral fellow of the Ministry of Health. BGPN is a MEC-FPU program fellow.

SUMMARY 1 The aim of the present study was to perform an evolutionary analysis of the morphometrical, biochemical and functional parameters of centriacinar emphysema induced by cadmium chloride (CdCl2) in rats and to determine the effects of concomitant N-acetylcysteine (NAC) administration. 2 Male Wistar rats were instilled orotracheally with either CdCl2 (n = 24) or saline (n = 24). One group of rats, consisting of both CdCl2- and saline-treated rats, was fed a normal diet (n = 24), whereas the other group received NAC (n = 24). 3 Changes in inspiratory capacity (IC), lung compliance (CL), expiratory flow at 75% (F75), forced vital capacity (FVC) and hydroxyproline content were assessed 2, 8, 21 and 45 days after instillation. Polymorphonuclear cells were evaluated 2 and 8 days after instillation and the mean linear intercept (Lm) was determined at 21 and 45 days. 4 Over time, CdCl2 instillation causes several changes that are bound up with centriacinar emphysema. The concomitant administration of NAC to CdCl2-treated rats partially reversed Lm at 21 days compared with CdCl2 alone (115 ± 2 vs 127 ± 2, respectively; P

This study focusses on the gene expression profile related to a new rhBMP-2 carrier material, chitosan film. This film could be suitable for use as an osteoinductive coating of commercially available titanium implants. The developed material was characterized, biocompatibility was tested and the cellular response was extensively characterized by transcriptional expression studies. Finally, in vivo studies were carried out to confirm the osteoinductivity of the developed coating. Results show good material properties for cell adhesion and proliferation. Presented data show cellular differentiation to the osteoblastic phenotype due to rhBMP-2, with a 90% common transcriptional response between the control rhBMP-2 treatment and the developed chitosan/rhBMP-2 film. The growing surface also had an influence on the observed cellular response and was quantified as 7% of the total. These results indicate that both the growth factor and the material induce a cell response, but this is mainly driven by the osteoinductor factor. In vivo, new bone formation and early vascularization was observed around chitosan/rhBMP-2 coated titanium pieces implanted in mouse muscle. In contrast, control implants did not induce this reaction. This work, therefore, shows both in vitro and in vivo that chitosan/rhBMP-2 film is a promising osteoinductive coating for titanium implantable materials.

International audience; Magnetic composites of Fe-based nanoparticles encapsulated in carbon/silica (C/SiO2@Fe) or carbon (C@Fe) matrices were prepared by laser-induced pyrolysis of aerosols. The powders were dispersed in aqueous solutions at pH 7 resulting in biocompatible colloidal dispersions with a high resistance to biodegradation. Structural and magnetic properties and the suitability of aqueous dispersions as contrast agent for MRI were analyzed. The results of these characterizations and the NMR relaxivity data are very encouraging for application of laser pyrolysis products in the field of living tissues.

The purpose of the present study was to compare neurotransmitter amino acid concentrations in the cerebrospinal fluid (CSF) and CSF/plasma ratios in 21 patients with dementia with Lewy bodies (DLB) and 26 matched controls. To this purpose, we used an ion-exchange chromatographic method. DLB patients exhibit higher CSF concentrations of asparagine (+25%) and glycine (+21%) compared to a control group, whereas no differences in CSF/plasma ratios were found between both groups. On the other hand, no alterations in concentrations of glutamate, aspartate and GABA were detected in CSF of patients compared to a control group. There was no correlation among amino acid levels and CSF/plasma ratios with age, age of onset, body mass index, duration of the disease or scores of the Mini Mental State Examination, UPDRS and Hoehn and Yahr stage. These results suggest a possible role of glycine and asparagine in the pathogenic mechanism of dementia with Lewy bodies.

The role of cannabinoid CB(1) receptors in the action of anxiolytics was examined. Deletion of CB(1) receptors resulted in increased anxiety-like behaviours in light/dark box, elevated plus maze and social interaction tests. Mutant mice presented basal low corticosterone concentrations and low proopiomelanocortin gene expression in the anterior lobe of the pituitary gland compared to wild-type mice. Ten minutes of restraint stress resulted in a twofold increase in corticosterone concentrations in the plasma of mutant mice, compared to wild-type mice. Bromazepam (50 or 100 microg/kg) markedly increased the time spent in light area in wild-type animals, though both doses were without effect in mutant mice. Administration of buspirone (1 or 2 mg/kg) produced anxiolytic effects in wild-type mice. In contrast, only the highest dose of buspirone had anxiolytic results in mutant mice. Our findings reveal that CB(1) receptors are involved in the regulation of emotional responses, and play a pivotal role in the action mechanism of anxiolytics. They suggest that alterations in the functional activity of the CB(1) receptor may be related to the emergence of anxiety disorders, and may affect treatment with anxiolytics.

Cigarette smoke (CS)-induced emphysema is a major component of chronic obstructive pulmonary disease (COPD). COPD treatment is based on the administration of bronchodilators and corticosteroids to control symptoms and exacerbations, however, to date, there are no effective therapies to reverse disease progression. Liver growth factor (LGF) is an albumin-bilirubin complex with mitogenic properties, whose therapeutic effects have previously been reported in a model of emphysema and several rodent models of human disease. To approach the therapeutic effect of LGF in a model of previously established emphysema, morphometric and lung function parameters, matrix metalloproteinase (MMP) activity and the expression of several markers, such as VEGF, PCNA, 3NT and Nrf2, were assessed in air-exposed and CS-exposed C57BL/6J male mice with and without intraperitoneal (i.p.) injection of LGF. CS-exposed mice presented a significant enlargement of alveolar spaces, higher alveolar internal area and loss of lung function that correlated with higher MMP activity, higher expression of 3NT and lower expression of VEGF. CS-exposed mice injected with LGF, showed an amelioration of emphysema and improved lung function, which correlated with lower MMP activity and 3NT expression and higher levels of VEGF, PCNA and Nrf2. Taken together, this study suggests that LGF administration ameliorates CS-induced emphysema, highlights the ability of LGF to promote alveolar cell proliferation and may be a promising strategy to revert COPD progression.