Abordaje endoscópico extendido del seno frontal: Experiencia en Hospital Clínico Universidad de Chile / Extended endoscopic approach of the frontal sinus: Experience at Hospital Clínico Universidad de Chile

Introducción: El seno frontal es una estructura compleja y desafiante en términos quirúrgicos, siendo descritas numerosas técnicas para su abordaje. Dentro de ellas se destaca el abordaje endoscópico extendido de seno frontal: Draf IIB y Draf III, como una importante alternativa para resolución de patología refractaria de seno frontal. Objetivo: Describir las características de pacientes sometidos a abordaje endoscópico extendido de seno frontal en Hospital Clínico Universidad de Chile (HCUCH). Material y Método: Estudio retrospectivo, descriptivo. Se incluyeron a pacientes sometidos a abordaje endoscópico extendido de seno frontal entre los años 2013 y 2021. Se analizaron variables clínicas, intraoperatorias y de seguimiento. Resultados: Se registraron 118 pacientes, de los cuales 64 cumplieron criterios de inclusión al estudio, con una edad promedio de 48 años. La patología más frecuente fue la rinosinusitis crónica poliposa (42%) seguido del mucocele (20%). Del total de pacientes, el 68% fue sometido a cirugía Draf IIB y el resto a Draf III. Todos los pacientes fueron estudiados con endoscopía e imágenes, y seguidos con parámetros clínicos y endoscópicos. El porcentaje de estenosis postoperatoria se estimó en 10%. Conclusión: El abordaje endoscópico nasal extendido figura como una alternativa útil para manejo de patología de seno frontal refractario a tratamiento. En nuestra experiencia las indicaciones, tipos de cirugía y tasa de complicaciones son concordantes con la literatura internacional.

Introduction: The frontal sinus is a complex and challenging structure in surgical terms, numerous techniques have been described for its approach, among them the extended endoscopic approach: Draf IIB and Draf III, figures as an important alternative for the resolution of refractory pathology of frontal sinus. Aim: To describe the characteristics of patients who underwent an extended endoscopic approach to the frontal sinus at the Hospital Clínico Universidad de Chile (HCUCH). Material and Method: A retrospective, descriptive study included patients who underwent an extended endoscopic approach to the frontal sinus between 2013 and 2021. Clinical, intraoperative, and follow-up variables were analyzed. Results: 118 patients were registered, of which 64 met the inclusion criteria for the study, with an average age of 48 years. The most frequent pathology was chronic polypous rhinosinusitis (42%), followed by mucocele (20%). Of the patients, 68% underwent Draf IIB surgery, while the rest received a Draf III type procedure. All patients were studied with endoscopy and images and followed up with clinical and endoscopic parameters. The percentage of post operatory stenosis was 10%. Conclusion: The extended nasal endoscopic approach appears as a valuable alternative for managing frontal sinus pathology refractory to treatment. In our experience, the indications, types of surgery, and rate of complications are consistent with the international literature.

[Contribution of « GIGA-Cancer ¼ at ULiege to the research on tumour microenvironment]. / Apport du «GIGA-Cancer¼ de l'ULiège à la recherche sur le microenvironnement tumoral.

The evolution of cancers is dictated by the intrinsic characteristics of malignant cells, but also by the multiple dynamic and reciprocal interactions that they establish with their tissue and cellular environment. This tumour microenvironment is therefore the subject of an ever-increasing part of cancer researches. These notably shed light on the plasticity of function of these non-malignant cells and on the diversity of their impact on the progression of the disease, both in primary tumours and during the formation of metastases. The improvement of the current therapy and the development of innovative treatments therefore require the identification of these cell subpopulations, either «allies¼ or «enemies¼ of aggressive cancer cells, as well as a more extensive understanding of the mechanisms modulating their phenotypes. This article summarizes some research projects carried out in two GIGA-Cancer laboratories supported by «Télévie¼ and the «Fondation Léon Frédéricq¼.

L'évolution de la pathologie cancéreuse est dictée par les caractéristiques intrinsèques des cellules malignes, mais également par les multiples interactions dynamiques et réciproques qu'elles établissent avec leur environnement tissulaire et cellulaire. Ce microenvironnement tumoral est donc l'objet d'une part sans cesse croissante des recherches en cancérologie. Celles-ci ont, notamment, mis en lumière la plasticité de fonction de ces cellules non malignes et la diversité de leurs impacts sur l'évolution de la maladie, tant dans les tumeurs primaires que lors de la formation des métastases. L'amélioration des traitements actuels et la mise au point de traitements novateurs nécessiteront donc l'identification fine de ces sous-populations cellulaires «alliées¼ ou «ennemies¼ des cellules cancéreuses agressives, ainsi qu'une compréhension accrue des mécanismes modulant leurs phénotypes. Cet article résume quelques projets de recherche menés dans deux laboratoires du GIGA-Cancer, soutenus notamment par Télévie et la Fondation Léon Fredericq.

Asthma-related inflammation promotes lung metastasis of breast cancer cells through CCL11-CCR3 pathway.

BACKGROUND: Mechanisms that preclude lung metastasis are still barely understood. The possible consequences of allergic airways inflammation on cancer dissemination were studied in a mouse model of breast cancer. METHODS: Balb/c mice were immunized and daily exposed to ovalbumin (OVA) from day 21. They were subcutaneously injected with 4T1 mammary tumor cells on day 45 and sacrificed on day 67. Lung metastases were measured by biophotonic imaging (IVIS® 200 Imaging System) and histological measurement of tumor area (Cytomine software). Effects of CCL11 were assessed in vivo by intratracheal instillations of recCCL11 and in vitro using Boyden chambers. CCR3 expression on cell surface was assessed by flow cytometry. RESULTS: The extent of tumor metastases was significantly higher in lungs of OVA-exposed mice and increased levels of CCL11 expression were measured after OVA exposure. Migration of 4T1 cells and neutrophils was stimulated in vitro and in vivo by recCCL11. 4T1 cells and neutrophils express CCR3 as shown by flow cytometry and a selective CCR3 antagonist (SB-297006) inhibited the induction of 4T1 cells migration and proliferation in response to recCCL11. CONCLUSIONS: Allergic inflammation generated by exposure to allergens triggers the implantation of metastatic cells from primary breast tumor into lung tissues plausibly in a CCL11-CCR3-dependent manner. This indicates that asthma related inflammation in lungs might be a risk factor for lung metastasis in breast cancer patients.

Spontaneous atopic dermatitis due to immune dysregulation in mice lacking Adamts2 and 14.

Since its first description, ADAMTS14 has been considered as an aminoprocollagen peptidase based on its high similarity with ADAMTS3 and ADAMTS2. As its importance for procollagen processing was never experimentally demonstrated in vivo, we generated Adamts14-deficient mice. They are healthy, fertile and display normal aminoprocollagen processing. They were further crossed with Adamts2-deficient mice to evaluate potential functional redundancies between these two highly related enzymes. Initial characterizations made on young Adamts2-Adamts14-deficient animals showed the same phenotype as that of Adamts2-deficient mice, with no further reduction of procollagen processing and no significant aggravation of the structural alterations of collagen fibrils. However, when evaluated at older age, Adamts2-Adamts14-deficient mice surprisingly displayed epidermal lesions, appearing in 2 month-old males and later in some females, and then worsening rapidly. Immunohistological evaluations of skin sections around the lesions revealed thickening of the epidermis, hypercellularity in the dermis and extensive infiltration by immune cells. Additional investigations, performed on young mice before the formation of the initial lesions, revealed that the primary cause of the phenotype was not related to alterations of the epidermal barrier but was rather the result of an abnormal activation and differentiation of T lymphocytes towards a Th1 profile. However, the primary molecular defect probably does not reside in the immune system itself since irradiated Adamts2-Adamts14-deficient mice grafted with WT immune cells still developed lesions. While originally created to better characterize the common and specific functions of ADAMTS2 and ADAMTS14 in extracellular matrix and connective tissues homeostasis, the Adamts2-Adamts14-deficient mice revealed an unexpected but significant role of ADAMTS in the regulation of immune system, possibly through a cross-talk involving mesenchymal cells and the TGFß pathways.

Differential impact of Limnoperna fortunei-herbicide interaction between Roundup Max® and glyphosate on freshwater microscopic communities.

Multiple anthropogenic stressors act simultaneously on the environment, with consequences different from those caused by single-stressor exposure. We investigated how the combination of the invasive mussel Limnoperna fortunei and a widely applied herbicide, Roundup Max®, affected freshwater microscopic communities and water quality. Further, we compared these results with those induced by the combination of the mussel and technical-grade glyphosate. We carried out a 34-day experiment in outdoor mesocosms, applying the following six treatments: 6 mg L(-1) of technical-grade glyphosate (G), the equivalent concentration of glyphosate in Roundup Max® (R), 100 mussels (M), the combination of mussels and herbicide either in the technical-grade or formulated form (MG and MR, respectively), and control (C). Herbicides significantly increased total phosphorus in water; R and MR showed greater initial total nitrogen and ammonium. R increased picoplankton abundance and caused an eightfold increase in phytoplankton, with high turbidity values; G had a lower effect on these variables. Herbicide-mussel combination induced an accelerated dissipation of glyphosate in water (MG 6.36 ± 0.83 mg G g DW(-1) day(-1) and MR 5.16 ± 1.26 mg G g DW(-1) day(-1)). A synergistic effect on ammonium was observed in MR but not in MG. MR and MG had an antagonistic effect on phytoplankton, which showed a drastic reduction due to grazing, as revealed by M. We provide evidence of differential effects of Roundup Max® and technical-grade glyphosate over water quality and microscopic communities, and in combination with mussels. However, in the combination of mussels and herbicides, mussels seem to play a leading role. In the presence of L. fortunei, the effects of higher nutrient availability provided by herbicides addition were counteracted by the filtration activity of mussels, which released nutrients, grazed on picoplankton and phytoplankton, and boosted the development of other primary producers, periphyton and metaphyton.

[Spiolto Respimat® : tiotropium-olodaterol fixed combination for therapy of chronic obstructive pulmonary disease (COPD)]. / Le médicament du mois. Spiolto Respimat® : combinaison fixe tiotropium-olodatérol pour le traitement de la broncho-pneumopathie chronique obstructive (BPCO).

Chronic Obstructive Pulmonary Disease (COPD) is a chronic airway disease that can be prevented and treated. The recommendations for therapy include bronchodilators from two classes (LAMA (Long Acting Muscarinic Antagonists) and LABA (Long Acting Beta2 Agonists)). Spiolto Respimat® is a LAMA/LABA combination therapy and comprises tiotropium (Spiriva®) and olodaterol (a LABA). Clinical studies show that Spiolto Respimat® is able to improve lung function tests (Increased FEV1, decreased hyperinflation and residual volume) and quality of life as compared to tiotropium or olodaterol. Studies also suggest that COPD exacerbations are decreased in patients treated by Spiolto Respimat® as compared to those treated with olodaterol (results of studies with exacerbations as primary outcome are not available yet). Safety of Spiolto Respimat® appears similar to tiotropium or olodaterol. Spiolto Respimat® indications is maintenance therapy for COPD. Reimbursement in Belgium requires that patients still display symptoms although already treated and that he/she has been previously treated by a LAMA.

La bronchopneumopathie chronique obstructive (BPCO) est une maladie chronique des voies aériennes qui est évitable et soignable. Le schéma de prise en charge comprend, entre autres, la prescription de bronchodilatateurs de la classe des LAMA (Long Acting Muscarinic Antagonists) et des LABA (Long Acting Beta2 Agonists). Le Spiolto Respimat® combine le tiotropium (un LAMA commercialisé sous le nom de Spiriva®) et l'olodatérol qui est un LABA. Les études cliniques montrent que le Spiolto Respimat® permet d'améliorer les indices de fonction respiratoire (augmentation du VEMS, diminution de l'hyperinflation et du volume résiduel) et d'améliorer la qualité de vie par rapport au tiotropium ou à l'olodatérol utilisés seuls. Le nombre d'exacerbations de la BPCO est également moindre chez les patients traités par Spiolto Respimat® par rapport à un traitement par olodatérol en monothérapie, mais des études dédiées à l'observation des exacerbations sont actuellement toujours en cours. La sécurité d'utilisation du Spiolto Respimat® est similaire à celle des deux mono-composants. Le Spiolto Respimat® est indiqué pour le traitement d'entretien de patients atteints de BPCO. Il est remboursé en Belgique chez les patients qui restent symptomatiques dans le cadre de la BPCO et qui sont déjà traités par un LAMA.

Additional effects of homeopathy on chronic periodontitis: a 1-year follow-up randomized clinical trial.

BACKGROUND AND OBJECTIVE: The aim of this study was test the hypothesis that homeopathy (H) enhances the effects of scaling and root planing (SRP) in patients with chronic periodontitis (CP). MATERIALS AND METHODS: 50 patients with CP were randomly allocated to one of two treatment groups: SRP (C-G) or SRP + H (H-G). Assessments were made at baseline and after 3 and 12 months of treatments. The local and systemic responses to the treatments were evaluated by clinical and serologic parameters, respectively. RESULTS: Both groups displayed significant improvements, however, using clinical attachment gain and reductions in HDL, LDL and Total Cholesterol, Triglycerides, Glucose and Uric acid, from baseline to 1 year, as criteria for treatment success, H-G performed significantly better than C-G. CONCLUSION: The findings of this 1-year follow-up randomized clinical trial suggest that homeopathic medicines, as an adjunctive to SRP, can provide significant local and systemic improvements for CP patients.

Oxidative stress-mediated iNKT-cell activation is involved in COPD pathogenesis.

Chronic obstructive pulmonary disease (COPD) is a major clinical challenge mostly due to cigarette smoke (CS) exposure. Invariant natural killer T (iNKT) cells are potent immunoregulatory cells that have a crucial role in inflammation. In the current study, we investigate the role of iNKT cells in COPD pathogenesis. The frequency of activated NKT cells was found to be increased in peripheral blood of COPD patients relative to controls. In mice chronically exposed to CS, activated iNKT cells accumulated in the lungs and strongly contributed to the pathogenesis. The detrimental role of iNKT cells was confirmed in an acute model of oxidative stress, an effect that depended on interleukin (IL)-17. CS extracts directly activated mouse and human dendritic cells (DC) and airway epithelial cells (AECs) to trigger interferonγ and/or IL-17 production by iNKT cells, an effect ablated by the anti-oxidant N-acetylcystein. In mice, this treatment abrogates iNKT-cell accumulation in the lung and abolished the development of COPD. Together, activation of iNKT cells by oxidative stress in DC and AECs participates in the development of experimental COPD, a finding that might be exploited at a therapeutic level.

Myeloid hypoxia-inducible factor 1α prevents airway allergy in mice through macrophage-mediated immunoregulation.

Hypoxia-inducible factor (HIF) has important roles in promoting pro-inflammatory and bactericidal functions in myeloid cells. Conditional genetic ablation of its major subunit Hif1α in the myeloid lineage consequently results in decreased inflammatory responses in classical models of acute inflammation in mice. By contrast, we report here that mice conditionally deficient for Hif1α in myeloid cells display enhanced sensitivity to the development of airway allergy to experimental allergens and house-dust mite antigens. We support that upon allergen exposure, MyD88-dependent upregulation of Hif1α boosts the expression of the immunosuppressive cytokine interleukin (IL)-10 by lung interstitial macrophages (IMs). Hif1α-dependent IL-10 secretion is required for IMs to block allergen-induced dendritic cell activation and consequently for preventing the development of allergen-specific T-helper cell responses upon allergen exposure. Thus, this study supports that, in addition to its known pro-inflammatory activities, myeloid Hif1α possesses immunoregulatory functions implicated in the prevention of airway allergy.

Curcumin-cyclodextrin complexes potentiate gemcitabine effects in an orthotopic mouse model of lung cancer.

BACKGROUND: Overall clinical outcome for advanced lung cancer remains very disappointing despite recent advances in treatment. Curcumin has been reported as potentially active against cancer. METHODS: Owing to poor curcumin solubility, we have used cyclodextrins (CD) as an excipient allowing a considerable increase of aqueous solubility and bioavailability of curcumin. The effects of solubilised curcumin have been evaluated in cell cultures as well as in an in vivo orthotopic lung tumour mouse model. RESULTS: Cell proliferation was reduced while apoptosis rates were increased when lung epithelial tumour cells were cultured in the presence of curcumin-CD complexes. For in vivo experiments, cells were grafted into lungs of C57Bl/6 mice treated by an oral administration of a non-soluble form of curcumin, CDs alone or curcumin-CD complexes, combined or not with gemcitabine. The size of orthotopically implanted lung tumours was reduced upon curcumin complex administration as compared with treatments with placebo or non-solubilised curcumin. Moreover, curcumin potentiated the gemcitabine-mediated antitumour effects. CONCLUSION: Our data demonstrate that curcumin, when given orally in a CD-solubilised form, reduces lung tumour size in vivo. In vitro experiments show impaired tumour cell proliferation and increased cell apoptosis. Moreover, our data underline a potential additive effect of curcumin with gemcitabine thus providing an efficient therapeutic option for antilung cancer therapy.

[Hymenoptera venom anaphylaxis and in-vitro diagnostic tools: new approaches to manage patients at high risk of a severe reaction]. / Allergie au venin d'hyménoptère et tests de laboratoires: de nouvelles approches pour la prise en charge des patients a haut risque de réaction sévère.

The consequences of Hymenoptera venom anaphylaxis are very severe but it is not obvious to predict which reactions will occur in one single patient when he is stung for the second time. For a couple of years, many new laboratory tests have been experimented and many studies published. CAST, BAT, WB, tryptase, sIgE and sIgG4 are the new valuable additional diagnostic tools that can help the decision to perform an immunotherapy or to discontinue this therapy after 3 years. The aim of our study was to determine which could be the profile of a desensitized patient and to screen for good candidates for venom immunotherapy.

The metalloproteinase ADAM-12 regulates bronchial epithelial cell proliferation and apoptosis.

OBJECTIVES: The ADAMs (a disintegrin and metalloproteinase) enzymes compose a family of membrane-bound proteins characterized by their multi-domain structure and ADAM-12 expression is elevated in human non-small cell lung cancers. The aim of this study was to investigate the roles played by ADAM-12 in critical steps of bronchial cell transformation during carcinogenesis. MATERIALS AND METHODS: To assess the role of ADAM-12 in tumorigenicity, BEAS-2B cells were transfected with a plasmid encoding human full-length ADAM-12 cDNA, and then the effects of ADAM-12 overexpression on cell behaviour were explored. Treatment of clones with heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) neutralizing antibodies as well as an EGFR inhibitor allowed the dissection of mechanisms regulating cell proliferation and apoptosis. RESULTS: Overexpression of ADAM-12 in BEAS-2B cells promoted cell proliferation. ADAM-12 overexpressing clones produced higher quantities of HB-EGF in their culture medium which may rely on membrane-bound HB-EGF shedding by ADAM-12. Targeting HB-EGF activity with a neutralizing antibody abrogated enhanced cell proliferation in the ADAM-12 overexpressing clones. In sharp contrast, targeting of amphiregulin, EGF or transforming growth factor-alpha failed to influence cell proliferation; moreover, ADAM-12 transfectants were resistant to etoposide-induced apoptosis and the use of a neutralizing antibody against HB-EGF activity restored rates of apoptosis to be similar to controls. CONCLUSIONS: ADAM-12 contributes to enhancing HB-EGF shedding from plasma membranes leading to increased cell proliferation and reduced apoptosis in this bronchial epithelial cell line.

Analysis of posttransplant diabetes mellitus prevalence in a population of kidney transplant recipients.

AIM: The onset of posttransplant diabetes mellitus (PTDM) among kidney recipients is associated with an increased risk of graft failure and death. Minimizing the risk of PTDM is a priority for long-term improvement in survival rates. We sought to evaluate the prevalence of PTDM and impaired fasting glucose (IFG) among a population of kidney transplant recipients to identify the risk factors and to evaluate graft and patient survivals. METHODS: We analyzed 250 consecutive Caucasian patients who received kidney allografts in our center between May 2000 and December 2005, with a median follow-up of 32 months (range, 1-78 months). RESULTS: We observed altered glucose metabolism in 17% of patients; specifically, the prevalences of PTDM and IFG were 12.2% and 4.8%, respectively. Patients who developed PTDM or IFG were overweight (BMI, 26.4+/-3.4 and 28.1+/-3.4 kg/m(2), respectively), whereas the normal glucose (NG) group's BMI was 23.8+/-3.5 kg/m(2) (P= .002 and P= .004, respectively). Prevalence of acute rejection was higher in the PTDM and IFG patients compared with the NG patients (60.7%, 63.6%, and 32.1%, respectively; P= .006; P< .04), while no difference was observed in terms of graft and patient overall survival. CONCLUSION: In our series of patients, we showed that being overweight represents a major risk factor for the development of PTDM, which results in an increased acute rejection rate. These results confirmed the importance of appropriate weight control among patients undergoing kidney transplantation, which should also be strictly monitored for all risk factors associated with the development of impaired glucose metabolism.

Emerging roles of ADAM and ADAMTS metalloproteinases in cancer.

A disintegrin and metalloproteinases (ADAMs) are a recently discovered family of proteins that share the metalloproteinase domain with matrix metalloproteinases (MMPs). Among this family, structural features distinguish the membrane-anchored ADAMs and the secreted ADAMs with thrombospondin motifs referred to as ADAMTSs. By acting on a large panel of membrane-associated and extracellular substrates, they control several cell functions such as adhesion, fusion, migration and proliferation. The current review addresses the contribution of these proteinases in the positive and negative regulation of cancer progression as mainly mediated by the regulation of growth factor activities and integrin functions.

Expression of a disintegrin and metalloprotease (ADAM and ADAMTS) enzymes in human non-small-cell lung carcinomas (NSCLC).

A Disintegrin and Metalloprotease (ADAM) are transmembrane proteases displaying multiple functions. ADAM with ThromboSpondin-like motifs (ADAMTS) are secreted proteases characterised by thrombospondin (TS) motifs in their C-terminal domain. The aim of this work was to evaluate the expression pattern of ADAMs and ADAMTS in non small cell lung carcinomas (NSCLC) and to investigate the possible correlation between their expression and cancer progression. Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and immunohistochemical analyses were performed on NSCLC samples and corresponding nondiseased tissue fragments. Among the ADAMs evaluated (ADAM-8, -9, -10, -12, -15, -17, ADAMTS-1, TS-2 and TS-12), a modulation of ADAM-12 and ADAMTS-1 mRNA expression was observed. Amounts of ADAM-12 mRNA transcripts were increased in tumour tissues as compared to the corresponding controls. In sharp contrast, ADAMTS-1 mRNA levels were significantly lower in tumour tissues when compared to corresponding nondiseased lung. These results were corroborated at the protein level by Western blot and immunohistochemistry. A positive correlation was observed between the mRNA levels of ADAM-12 and those of two vascular endothelial growth factor (VEGF)-A isoforms (VEGF-A(165) and VEGF-A(121)). Taken together, these results providing evidence for an overexpression of ADAM-12 and a lower expression of ADAMTS-1 in non-small-cell lung cancer suggest that these proteases play different functions in cancer progression.

Elevated MMP-12 protein levels in induced sputum from patients with COPD.

BACKGROUND: Several matrix metalloproteinases (MMPs) are involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). In mice, MMP-12 plays a crucial role in the development of cigarette smoke induced emphysema. A study was undertaken to investigate the role of MMP-12 in the development of COPD in human smokers. METHODS: Induced sputum samples were collected from patients with stable COPD (n = 28), healthy smokers (n = 14), never smokers (n = 20), and former smokers (n = 14). MMP-12 protein levels in induced sputum were determined by ELISA and compared between the four study groups. MMP-12 enzymatic activity in induced sputum was evaluated by casein zymography and by cleaving of a fluorescence quenched substrate. RESULTS: Median (IQR) MMP-12 levels were significantly higher in COPD patients than in healthy smokers, never smokers, and former smokers (17.5 (7.1-42.1) v 6.7 (3.9-10.4) v 4.2 (2.4-11.3) v 6.1 (4.5-7.6) ng/ml, p = 0.0002). MMP-12 enzymatic activity was significantly higher in patients with COPD than in controls (4.11 (1.4-8.0) v 0.14 (0.1-0.2) microg/microl, p = 0.0002). CONCLUSION: MMP-12 is markedly increased in induced sputum from patients with stable COPD compared with controls, suggesting a role for MMP-12 in the development of COPD in smokers.

Automated method for the determination of a new matrix metalloproteinase inhibitor in ovine plasma and serum by coupling of restricted access material for on-line sample clean-up to liquid chromatography.

A fully automated liquid chromatographic method was developed for the determination of Ro 28-2653, a new synthetic inhibitor of matrix metalloproteinases (MMPs), in ovine serum and plasma. The method was based on the coupling of a pre-column packed with restricted access material, namely LiChrospher RP-8 ADS (alkyl diol silica), for sample clean-up to an analytical column containing octyl silica stationary phase. One hundred microl of biological sample, to which 2-propanol was automatically added, were injected onto the ADS pre-column, which was then washed with a washing liquid consisting of a mixture of 25 mM phosphate buffer (pH 7.0) and acetonitrile (90:10; v/v) for 10 min. By rotation of the switching valve, the analyte was then eluted in the back-flush mode with the LC mobile phase composed of a mixture of acetonitrile and 25 mM phosphate buffer (pH 7.0) (57:43; v/v). The UV detection was performed at 395 nm. The main parameters likely to influence the sample preparation technique were investigated. The method was then validated over a concentration range from 17.5 to 1950 ng/ml, the first concentration level corresponding to the lower limit of quantitation. At this concentration level, the mean bias and the R.S.D. value for intermediate precision were -2.4% and 4.2%, respectively.

La mantención de peso en humanos podría ser equivalente a la restricción calórica de los modelos animales? / Weight maintenance in humans. Could it mimic calorie restriction of animal models?

BACKGROUND: Energy restriction (ER) extends life span in animals, by decreasing oxidative stress. AIM: To compare adiposity, metabolic variables and DNA oxidative damage, among adults, reporting a constant body weight (weight maintainers), versus those reporting a progressive increase (weight gainers). SUBJECTS AND METHODS: Clinical history, dietary recall, anthropometric measures, abdominal CT scan and fasting blood samples (to measure lipoproteins, glucose and insulin), were obtained in 44 males. These subjects were classified as weight maintainers if they had a change in weight of 3 kg or less in the last 10 years, or weight gainers, if they had a weight increment of more than 6 kg, in the same lapse. Oxidative damage was assessed by 8-hydroxydeoxyguanosine (8-OHdG), in DNA extracted from circulating lymphocytes, in 5 weight maintainers, 8 weight gainers and 5 healthy elders. RESULTS: Energy Intake was 18% higher in weight gainers (p <0.01). Adiposity and central fat were higher among weight gainers (p <0.01). Abdominal fat correlated with serum lipoproteins, glucose and insulin sensitivity, assessed by the Homeostasis Model Assessment (HOMA). 8-OHdG levels did not differ between groups. CONCLUSIONS: The analysis of weight change based on the clinical history correlates with actual body composition, thus it may be a reliable indicator of long term energy Intake. This method could be comparable to weight clamp models employed in animals to study aging.

Cyclodextrins as a potential carrier in drug nebulization.

The inhalation route is widely studied for many drug applications focusing on either local or systemic distributions. One matter of concern is the solubilization of hydrophobic drugs. We have studied the feasibility of using different cyclodextrins (CDs) to elaborate pharmaceutical formulations for the inhalation route and tested the short-term toxicity of such formulations administered by inhalation to C57BL/6 mice. We have shown that HP-beta-CD, gamma-CD, as well as RAMEB aqueous solutions can undergo aerosolization and that the resulting droplet-size ranges are compatible with pulmonary deposition. In vivo, we have demonstrated that short-term exposure to inhaled HP-beta-CD, gamma-CD and RAMEB solutions are non-toxic after assessing bronchoalveolar lavage (BAL), lung and kidney histology, bronchial responsiveness to methacholine and blood urea. The only change noted is a slight increase in lymphocyte count in the BAL after HP-beta-CD and gamma-CD inhalation. We conclude that CDs are useful in significantly enhancing the solubility of apolar drugs with a view to inhalation therapy although an increase in lymphocyte counts in the BAL after CDs inhalations needs further investigations.