Results in clinical practice in the treatment of severe eosinophilic asthma with mepolizumab: a real-life study.

OBJECTIVE: Add-on therapy with monoclonal antibodies is the recommended therapy for severe asthmatic patients refractory to maintenance treatment. In randomized control trials, mepolizumab reduced the number of exacerbations, the need of oral corticosteroids (OCS), increased asthma control, and lung function in a population of uncontrolled severe eosinophilic asthmatic patients. In this piece of work, we aimed to assess mepolizumab efficacy and safety in a cohort of patients with severe eosinophilic asthma in real-life conditions. METHODS: A retrospective study was carried out at eight hospitals from Asturias (Spain). The sample included patients treated with mepolizumab from 1 January 2016 to 31 March 2019. Demographic and clinical variables were collected, including OCS use, asthma control, lung function, and exacerbation rate. RESULTS: Sixty-nine patients (72% women) with mean age 56 ± 13 years were included. Annual exacerbation rate decreased from 4.7 (SD 3.7) to 1.3 (SD 2.5) (p < 0.001). The number of patients requiring OCS treatment decreased from 25 patients (36%, mean prednisone dose = 18 mg/day) to 13 patients (19%, mean prednisone dose = 9 mg/day) (p < 0.001). Twelve patients (48%) stopped OCS treatment. Forced expired volume in one second (FEV1) as percentage increased from 68% (SD 20) to 76% (SD 21) (p < 0.001). Fifty-six patients (81%) were considered responders to mepolizumab. No serious adverse events were detected during the study period. CONCLUSIONS: Overall, this study demonstrates mepolizumab efficacy and safety in a cohort of patients with uncontrolled severe eosinophilic asthma in routine clinical practice.

The role of macrophages and mast cells in lymphangiogenesis and angiogenesis in cervical carcinogenesis.

During carcinogenesis it is known that growth factors and cytokines from stromal and inflammatory cells from the microenvironment promote angiogenesis and lymphangiogenesis. However, the participation of macrophages and mast cells in these processes is not well understood. The aim of this study was to evaluate the relationship between mast cell and macrophage density with blood and lymphatic vessels in various stages of carcinoma of the uterine cervix. Tissue sections from archival paraffin-embedded samples from cases with cervical intraepithelial neoplasias (CIN) 1, 2, 3, carcinoma in situ, and invasive carcinoma were used. Immunohistochemical staining was done using the following antibodies: anti-LYVE-1; anti-CD31; anti-CD68, and anti-tryptase. Our results showed a significant increase in the number of macrophages in carcinoma in situ, a correlation between lymphatic vessels and macrophages in premalignant lesions CIN 2, and a correlation between mast cells and blood vessels in both CIN 2 and carcinoma in situ. In conclusion, our data underscore the importance of the recruitment of macrophages and mast cells in the development of tumor-associated blood and lymphatic capillaries.

O-GlcNAc-selective-N-acetyl-beta-D-glucosaminidase activity and mRNA expression in muscle is related to glucosamine-induced insulin resistance.

Glucosamine (GlcN)-induced insulin resistance is associated with an increase in O-linked-N-acetylglucosaminylated modified proteins (O-GlcNAcylated proteins). The role played by O-GlcNAc-selective-N-acetyl-beta-D-glucosaminidase (O-GlcNAcase), which removes O-N-acetyl-glucosamine residues from O-GlcNAcylated proteins, has not yet been demonstrated. We investigated whether GlcN-induced whole-body insulin resistance is related to tissue O-GlcNAcase activity and mRNA expression. GlcN (30 mumol/kg/min) or physiological saline (control) was intravenously infused into Sprague-Dawley rats for 2 h. After GlcN treatment, rats were subjected to the following: intravenous glucose tolerance test, insulin tolerance test or removal of the liver, muscle and pancreas. GlcN was found to provoke hyperglycemia compared to control (8.6 +/- 0.41 vs. 4.82 +/- 0.17 mM, p < 0.001). The insulin resistance index (HOMA-IR) increased (15.76 +/- 1.47 vs. 10.14 +/- 1.41, p < 0.001) and the beta-cell function index (HOMA-beta) diminished (182.69 +/- 22.37 vs. 592.01 +/- 103, p < 0.001). Liver glucose concentration was higher in the GlcN group than in the control group (0.37 +/- 0.04 vs. 0.24 +/- 0.038 mmol/g dry weight, p < 0.001). Insulin release index (insulin/glucose) was less in the GlcN group than in the control (2.2 +/- 0.1 vs. 8 +/- 0.8 at 120 min, p < 0.001). In the GlcN group, muscle O-GlcNAcase activity diminished (0.28 +/- 0.019 vs. 0.36 +/- 0.018 nmol of p-nitrophenyl/mg protein/min, p < 0.001), and K(m) increased (1.51 +/- 0.11 vs. 1.12 +/- 0.1 mM, p < 0.001) compared to the control. In the GlcN group, O-GlcNAcase activity/mRNA expression was altered (0.6 +/- 0.07 vs. 1 +/- 0.09 of control, p < 0.05). In conclusion, O-GlcNAcase activity is posttranslationally inhibited during GlcN-induced insulin resistance.

Changes in the glucose-6-phosphate dehydrogenase activity in granulosa cells during follicular atresia in ewes.

Apoptosis of granulosa cells during follicular atresia is preceded by oxidative stress, partly due to a drop in the antioxidant glutathione (GSH). Under oxidative stress, GSH regeneration is dependent on the adequate supply of NADPH by glucose-6-phosphate dehydrogenase (G6PD). In this study, we analyzed the changes of G6PD, GSH, and oxidative stress of granulosa cells and follicular liquid and its association with apoptosis during atresia of small (4-6 mm) and large (>6 mm) sheep antral follicles. G6PD activity was found to be higher in granulosa cells of healthy small rather than large follicles, with similar GSH concentration in both cases. During atresia, increased apoptosis and protein oxidation, as well as a drop in GSH levels, were observed in follicles of both sizes. Furthermore, the activity of G6PD decreased in atretic small follicles, but not in large ones. GSH decreased and protein oxidation increased in follicular fluid. This was dependent on the degree of atresia, whereas the changes in G6PD activity were based on the type of follicle. The higher G6PD activity in the small follicles could be related to granulosa cell proliferation, follicular growth, and a lower sensitivity to oxidative stress when compared with large follicles. The results also indicate that GSH concentration in atretic follicles depends on other factors in addition to G6PD, such as de novo synthesis or activity of other NADPH-producing enzymes. Finally, lower G6PD activity in large follicles indicating a higher susceptibility to oxidative stress associated to apoptosis progression in follicle atresia.

DD genotype of angiotensin-converting enzyme in type 2 diabetes mellitus with renal disease in Mexican Mestizos.

AIM: The DD genotype of angiotensin-converting enzyme (ACE) has been suggested as a major contributor of diabetic nephropathy in several populations. The purpose of the present study was to determine whether micro/macroalbuminuria is associated with ACE insertion/deletion (I/D) polymorphism in Mexican Mestizos with type 2 diabetes mellitus. METHODS: A total of 435 patients with type 2 diabetes mellitus, of whom 233 had albuminuria, were characterized for the ACE I/D polymorphism by the polymerase chain reaction method. RESULTS: Clinical and biochemical characteristics and frequencies according to DD, ID and II genotypes in patients with and without albuminuria showed no significant differences. However, only females with micro/macroalbuminuria showed higher frequency of a DD genotype than those without albuminuria (27.9%, 21.2% and 10.5%, respectively; P

[Cancer lymphangiogenesis and its role in metastasic dissemination]. / Linfangiogénesis en el cáncer y su papel en la diseminación metastásica.

It is well-known that there are different tumor-type-dependent metastatic patterns. For example, in carcinomas metastatic spread is preferentially via the lymphatic system by which they reach regional lymph nodes through pre-existent afferent lymph vessels and/or newly formed lymph capillaries; while in sarcomas the favored pathway is through bloodvessels. These metastatic patterns have been used for many years by clinicians and surgeons for staging and tumor resection, particularly in the case of breast cancer. Recently this knowledge has been applied to detection and resection of sentinel lymph nodes. The lymphatic system drains the interstitial fluid from tissues and reincorporates it into the blood flow; in addition, it forms part of the host's immune defense and in pathological conditions, induces different types of lymph edema and participates in tumor invasion and metastasis. Although, the study of lymphangiogenesis was stagnated for several decades, it was not until a few years ago that biomolecular mechanisms were discovered and many specific markers are now in use to study the process of tumor dissemination and metastasis. There is a tendency to utilize molecular knowledge in clinical settings for grading and estimating prognostic significance of tumors as well as to develop specific therapeutic strategies.

Crosstalk between coagulation and inflammation during Dengue virus infection.

Dengue fever is the most prevalent viral disease transmitted by vectors (Aedes aegypti, Aedes albopictus) in worldwide. More than 100 million cases occur annually with a mortality rate of 5% and no safe vaccine is available. The pathogenesis of Dengue, where host and viral factors participate in the establishment of Dengue haemorrhagic fever (DHF) and Dengue shock syndrome (DSS) remains unresolved. Clinical observations have revealed significant abnormalities in coagulation and inflammation systems, with increased levels of tissue factor (TF) and the chemokine IL-8, correlating with the severity of the disease and implicating damage to endothelial vascular cells (EVC). Here we present novel insights concerning the crosstalk between the regulatory signaling pathways of the coagulation-inflammation processes, during Dengue virus (DV) infection of EVC. We found that DV up-regulates Protease Activated receptor type-1 (inflammation) and TF (coagulation) receptors, via the phosphorylation of p38 and ERK1/2 MAPKs, which favor the activation of NF-kappaB transcription factor. This induces pro-inflammatory (IL-8) or pro-adhesive (VCAM-1) gene expression which may lead to EVC activation. The elucidation of the basic principles that signal these processes has important implications for the design of new therapeutic strategies for DHF/DSS.

Radiological changes due to atypical aneurysms of the subclavian artery / Alteración radiológica secundario a aneurismas aberrantes de arteria subclavia

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In vitro proliferation, expansion, and differentiation of a CD34+ cell-enriched hematopoietic cell population from human umbilical cord blood in response to recombinant cytokines.

BACKGROUND: The conditions and mechanisms that control the in vitro growth of hematopoietic stem/progenitor cells (contained within the population of CD34+ cells) are still not completely understood. METHODS: By using an immunomagnetic system, we have enriched for umbilical cord blood (UCB)-derived CD34+ cells (55% of total cells recovered vs. 0.8% of total cells prior to the enrichment procedure) and analyzed their in vitro growth (proliferation, expansion, and differentiation) in a liquid culture system in the absence or presence of different recombinant cytokine combinations. RESULTS: When the selected cells were cultured in the absence of recombinant cytokines, no proliferation or expansion was observed. In the presence of steel factor (SF) and interleukin-6 (IL-6), total cell number was increased nearly fourfold; however, no progenitor cell expansion took place. When cultures were supplemented with SF and IL-6 together with IL-3 and erythropoietin (EPO), a rapid proliferation of the CD34+ -enriched cell population was observed with a selective stimulation of erythropoiesis. However, this stimulation was only transient, suggesting that there was a rapid exhaustion of erythroid progenitor cells within the first 10 days. Significantly higher levels of proliferation and expansion of progenitor cells were observed in the presence of SF, IL-6, GM-CSF, and G-CSF with preferential stimulation of myelopoiesis. Interestingly, such stimulation of myelopoiesis was sustained for the entire culture period (>30 days). The highest levels of proliferation and expansion were observed in the presence of all six cytokines. Under these conditions, erythropoiesis was also sustained only transiently (10 days), whereas myelopoiesis was sustained for >30 days. CONCLUSIONS: This study indicates that significant proliferation and expansion of hematopoietic progenitors can be achieved in vitro when culturing a cell population in which CD34+ cells comprise only >50% of the total cells. Our results also suggest that myeloid progenitors (those responding to GM-CSF and G-CSF) possess higher expansion potentials in vitro than their erythroid counterparts. The methods described here for the enrichment and culture of CD34+ cells may be relevant in the development of protocols for the ex vivo proliferation and expansion of hematopoietic progenitors for transplantation.

Pneumothorax as atypical presentation of bronchial carcinoid.

We present a case of pneumothorax associated with an endobronchial carcinoid tumor in a 18-year-old man with dyspnea and chest pain. Additional tests were done, identifying in the chest roentgenogram a complete left pneumothorax with persistent leak, which was confirmed by computed tomography of the chest, and also finding an endobronchial lesion that limited the complete reexpansion of the left lung. Surgical excision was performed, and the lesion was identified as a typical bronchial carcinoid, with satisfactory outcome after the intervention.

[Outbreak of Coxiella burnetii pneumonia]. / Brote epidémico de neumonía por Coxiella burnetii.

OBJECTIVE: The aim of this study was to describe an outbreak of Q fever pneumonia in the North of Spain. MATERIAL AND METHODS: We report 60 cases of Coxiella burnetii pneumonia in a rural area with 16,000 inhabitants during January and February 2003. The diagnosis was serologically confirmed by indirect immunofluorescence. RESULTS: Mean age of the patients was 46 years and the male/female ratio was 2.5. Overall, 52% of the patients lived in urbanized areas and contact with animals was declared in only 7% of patients. Sixty percent of patients were hospitalized. The most frequent clinical presentation was an acute febrile process (100%) and headache (80%); respiratory symptoms were infrequent. The most common liver function alteration was elevated transaminase levels (35%). Radiologic evidence of a single lobar or segmental alveolar opacity was found in 87% of patients, and most commonly affected the lower lobes. Chest radiographs returned to normal in 70% of patients within the two first months. Empirical treatment consisted of beta-lactams and macrolides in 38%, macrolides alone in 8%, and quinolones in 23%; 27% were treated with beta-lactams alone. No deaths occurred. CONCLUSIONS: Coxiella burnetii infection is very prevalent in the north of Spain and should be suspected and investigated when outbreaks of a pneumonic illness occur.

Linfangiogénesis en el cáncer y su papel en la diseminación metastásica / Cancer lymphangiogenesis and its role in metastasic dissemination

Es bien sabido que existen diferentes patrones de metástasis dependiendo del tipo tumoral. Por ejemplo, la diseminación metastásica de los carcinomas es vía linfática preferencialmente, las células neoplásicas llegan a los ganglios linfáticos regionales a través de vasos linfáticos aferentes preexistentes o capilares linfáticos de nueva formación; en cambio, en los sarcomas la vía principal es a través de los vasos sanguíneos. Estos patrones metastásicos han sido utilizados durante muchos años por los clínicos y cirujanos para la etapificación y resección tumoral, particularmente en cáncer de mama. Recientemente este conocimiento ha sido aplicado para la detección y resección del ganglio centinela. El sistema linfático drena el líquido intersticial de los tejidos y lo reincorpora al sistema sanguíneo; además, forma parte de la defensa inmune del huésped y en condiciones patológicas induce diferentes tipos de linfedema y participa en la invasión y metástasis. El estudio de la linfangiogénesis permaneció aletargado por muchas décadas y no es sino hasta los últimos años que se han descrito mecanismos biomoleculares y marcadores específicos, los cuales actualmente se están utilizando para estudiar el proceso de diseminación tumoral y metástasis. Existe una tendencia hacia la aplicación clínica de este conocimiento molecular en la clínica para estimar el significado pronóstico de los tumores, así como para desarrollar estrategias terapéuticas específicas.

It is well-known that there are different tumor-type-dependent metastatic patterns. For example, in carcinomas metastatic spread is preferentially via the lymphatic system by which they reach regional lymph nodes through pre-existent afferent lymph vessels and/or newly formed lymph capillaries; while in sarcomas the favored pathway is through bloodvessels. These metastatic patterns have been used for many years by clinicians and surgeons for staging and tumor resection, particularly in the case of breast cancer. Recently this knowledge has been applied to detection and resection of sentinel lymph nodes. The lymphatic system drains the interstitial fluid from tissues and reincorporates it into the blood flow; in addition, it forms part of the host's immune defense and in pathological conditions, induces different types of lymph edema and participates in tumor invasion and metastasis. Although, the study of lymphangiogenesis was stagnated for several decades, it was not until a few years ago that biomolecular mechanisms were discovered and many specific markers are now in use to study the process of tumor dissemination and metastasis. There is a tendency to utilize molecular knowledge in clinical settings for grading and estimating prognostic significance of tumors as well as to develop specific therapeutic strategies.

Brote epidémico de neumonía por Coxiella burnetii / Outbreak of Coxiella burnetii pneumonia

Objetivo. El propósito del estudio es describir un brote epidémico de neumonía por Coxiella burnetii que ocurrió en un área rural del norte de España. Material y métodos. Se describen 60 casos de neumonía por C. burnetii en un área rural de 16.000 habitantes del norte de España durante enero y febrero de 2003. El diagnóstico fue confirmado serológicamente por inmunofluorescencia indirecta. Resultados. La edad media fue de 46 años siendo la relación varón/mujer de 2,5. El 52% de los pacientes vivían en un área urbana y solamente el 7% referían contacto con animales. Fueron hospitalizados el 60% de los pacientes. La clínica más frecuente fue un proceso febril agudo (100%) y cefalea (80%) con escasos síntomas respiratorios. La alteración hepática más frecuente fue la elevación de las transaminasas (35%). Las alteraciones radiológicas consistieron en una condensación alveolar lobular o segmentaria en el 87% de los pacientes siendo los lóbulos inferiores los más frecuentemente afectados. La radiografía de tórax se normalizó en el 70% de los pacientes en los primeros 2 meses. El tratamiento empírico utilizado con mayor frecuencia fueron los betalactámicos asociados a macrólidos en el 38%, solamente macrólidos en el 8% y quinolonas en el 23% de los casos. El 27% de los pacientes fueron tratados únicamente con betalactámicos. Ningún paciente falleció. Conclusiones. La infección por C. burnetii es muy prevalente en el norte de España y debe de ser sospechada e investigada en aquellos casos de brote epidémico neumónico (AU)

Objective. The aim of this study was to describe an outbreak of Q fever pneumonia in the North of Spain. Material and methods. We report 60 cases of Coxiella burnetii pneumonia in a rural area with 16,000 inhabitants during January and February 2003. The diagnosis was serologically confirmed by indirect immunofluorescence. Results. Mean age of the patients was 46 years and the male/female ratio was 2.5. Overall, 52% of the patients lived in urbanized areas and contact with animals was declared in only 7% of patients. Sixty percent of patients were hospitalized. The most frequent clinical presentation was an acute febrile process (100%) and headache (80%); respiratory symptoms were infrequent. The most common liver function alteration was elevated transaminase levels (35%). Radiologic evidence of a single lobar or segmental alveolar opacity was found in 87% of patients, and most commonly affected the lower lobes. Chest radiographs returned to normal in 70% of patients within the two first months. Empirical treatment consisted of beta-lactams and macrolides in 38%, macrolides alone in 8%, and quinolones in 23%; 27% were treated with beta-lactams alone. No deaths occurred. Conclusions. Coxiella burnetii infection is very prevalent in the north of Spain and should be suspected and investigated when outbreaks of a pneumonic illness occur (AU)