Angiogenesis is promoted by hypoxic cervical carcinoma-derived extracellular vesicles depending on the endothelial cell environment.

INTRODUCTION: Cancer needs perfusion for its growth and metastasis. Cancer cell-derived extracellular vesicles (CA-EVs) alter the tumor microenvironment (TME), potentially promoting angiogenesis. We hypothesize that conditions in the tumor, e.g., hypoxia, and in the target cells of the TME, e.g., nutrient deprivation or extracellular matrix, can affect the angiogenic potential of CA-EVs, which would contribute to explaining the regulation of tumor vascularization and its influence on cancer growth and metastasis. METHODS: CA-EVs were isolated and characterized from cervical carcinoma cell lines HeLa and SiHa cultured under normoxia and hypoxia, and their angiogenic potential was evaluated in vitro in three endothelial cells (ECs) lines and aortic rings, cultured in basal (growth factor-reduced) or complete medium. RESULTS: Hypoxia increased EV production 10-100 times and protein content 2-4 times compared to normoxic CA-EVs. HeLa-EVs contained six times more RNA than SiHa-EVs, and this concentration was not affected by hypoxia. Treatment with CA-EVs increased tube formation and sprouting in ECs and aortic rings cultured in basal medium and long-term stabilized the stablished vascular networks formed by ECs cultured in complete medium. CONCLUSION: Hypoxia differentially affects CA-EVs in a cell line-dependent manner. The cellular environment (nutrient availability and extracellular matrix scaffold) influences the effect of CA-EV on the angiogenic potential of ECs.

Endothelial plasticity across PTEN and Hippo pathways: A complex hormetic rheostat modulated by extracellular vesicles.

Vascularization is a multifactorial and spatiotemporally regulated process, essential for cell and tissue survival. Vascular alterations have repercussions on the development and progression of diseases such as cancer, cardiovascular diseases, and diabetes, which are the leading causes of death worldwide. Additionally, vascularization continues to be a challenge for tissue engineering and regenerative medicine. Hence, vascularization is the center of interest for physiology, pathophysiology, and therapeutic processes. Within vascularization, phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and Hippo signaling have pivotal roles in the development and homeostasis of the vascular system. Their suppression is related to several pathologies, including developmental defects and cancer. Non-coding RNAs (ncRNAs) are among the regulators of PTEN and/or Hippo pathways during development and disease. The purpose of this paper is to review and discuss the mechanisms by which exosome-derived ncRNAs modulate endothelial cell plasticity during physiological and pathological angiogenesis, through the regulation of PTEN and Hippo pathways, aiming to establish new perspectives on cellular communication during tumoral and regenerative vascularization.

Current Needs Assessment for Using Lung Clearance Index for Asthma in Clinical Practice.

PURPOSE OF REVIEW: Asthma pathophysiology has shown that remodeling of the bronchial airways mainly affects the small rather than large airways. The severity of asthma is conventionally measured by forced expiratory volume 1 (FEV1) but this maneuver is insensitive to changes in distal airways with smaller diameter. The aim of this review is to evaluate the current evidence supporting LCI as a clinical tool for assessing small airways disease in asthma patients, as well as whether it is useful as a treatment response parameter in severe therapy-resistant asthma (STRA) patients. RECENT FINDINGS: There is an increasing need for novel tests that can assess distal airway disease in asthma. Lung Clearance Index (LCI) may be a useful test for assessing more severe airway obstruction and the persistence of small airway disease. LCI measurement has been shown to be more sensitive than spirometry in cystic fibrosis (CF), but its clinical utility in asthma has not been thoroughly investigated. LCI abnormalities may be a sensitive marker for the persistence of small distal airway disease and may be associated with a more severe asthma endotype unresponsive to inhaled glucocorticoids. There is a need to identify other lung function tests for asthma that can identify early airway remodeling while simultaneously measuring the rate of lung function impairment. When compared to other conventional methods, multiple-breath washout (MBW) measures the lung clearance index (LCI), a more sensitive predictor of early airway disease that is feasible to perform in children. The goal of this review is to evaluate the current evidence of LCI as a clinical tool in asthma patients.

A new set of equations for the simplified calibration of fluorescence Ca2+ transients in skeletal muscle fibers.

The classical approach for calibrating non-ratiometric fluorescent Ca2+ dyes entails the measurement of the fluorescence maximum (Fmax) and minimum (Fmin), as well as the dissociation constant (Kd) of the Ca2+-Dye reaction (model 1). An alternative equation does not need the Fmin but requires the rate constants kon and koff (model 2). However, both approaches are experimentally time consuming and the rate constants for several dyes are unknown. Here, we propose a set of equations (model 3) that simplify the calibration of fluorescent Ca2+ transients obtained with non-ratiometric dyes. This equation allows the calibration of signals without using the Fmin: [Ca2+] = Kd(F - Frest/Fmax - F) + [Ca2+]IR(Fmax - Frest/Fmax - F), where [Ca2+]IR is the resting [Ca2+]. If the classical calibration approach is followed, the Fmin can be estimated from: Fmin = Frest - ([Ca2+]IR(Fmax - Frest)/Kd). We tested the models' performance using signals obtained from enzymatically dissociated flexor digitorum brevis fibers of C57BL/6 mice loaded with Fluo-4, AM. Model 3 performed the same as model 2, and both gave peak [Ca2+] values 15 ± 0.3% (n = 3) lower than model 1, when we used our experimental Fmin (1.24 ± 0.11 A.U., n = 4). However, when we used the mathematically estimated Fmin (6.78 ± 0.2 A.U) for model 1, the peak [Ca2+] were similar for all three models. This suggests that the dye leakage makes a correct determination of the Fmin unlikely and induces errors in the estimation of [Ca2+]. In conclusion, we propose simpler and time-saving equations that help to reliably calibrate cytosolic Ca2+ transients obtained with non-ratiometric fluorescent dyes. The use of the estimated Fmin avoids the uncertainties associated with its experimental measurement.

What kind of information and communication technologies do patients with COPD prefer to use? A cross-sectional study in Latin America.

The aim of the present study was to assess the frequency of uses and preferences of information and communication technologies (ICTs) among Latin American chronic obstructive pulmonary disease (COPD) patients. We conducted an anonymous cross-sectional survey study on Latin American COPD patients. The adapted version of the Michigan questionnaire was employed in eligible outpatients in different cities of Latin America. We categorized age and educational levels into three groups. The time passed since COPD diagnosis was categorized as ≤5 years and >5 years. χ2 and crude and adjusted logistic regressions were performed. A total of 256 patients were enrolled with a mean age of 68.7 years old. The most recurrently used ICTs were short message service (SMS; 47.1%) and WhatsApp (30.7%) for receiving COPD information. Moreover, SMS (85.8%) and Facebook (36.1%) were rated as useful for asking physicians information about COPD. Regression analysis showed that the best predictor for patients using ICTs, for any purpose, was higher education (undergraduate or graduate school). Understanding the preferences of ICTs among COPD patients could help improve patient's outcomes through developing applications in response to specific requirements of each patient.

Knowledge and management of chronic spontaneous urticaria in Latin America: a cross-sectional study in Ecuador.

BACKGROUND: The current EAACI/GA2LEN/EDF/WAO guideline for urticaria provide specific recommendations for the diagnostic workup and treatment of patients with chronic spontaneous urticaria (CsU). This study explored if physicians in Ecuador know these recommendations and implement them in their actual clinical practice for CsU. METHODS: We investigated physicians who treat CsU patients in a cross-sectional study using a standardized questionnaire. Descriptive statistics were employed, adjusted logistic regression was performed to assess the link of guideline knowledge and use of therapy. RESULTS: Seven hundred forty surveys were collected and analyzed. The mean age of physicians was 42.3 (±12.5) years. Most of the participants (65.1%) were general physicians (GP), 13.7% were pediatricians, 11.0% internists, 6.8% dermatologists or allergists (D/A). Only 18.8% knew the EAACI/GA2LEN/EDF/WAO guideline. 44.5% of GPs searched for CsU etiology in contrast to 90% of D/A. Most common diagnostic test was total serum IgE (83.5%). Most common first line symptomatic treatment was oral corticoids (46.3%), followed by second generation antihistamines (sgAHs, 36.8%). A/D prescribed more sgAHs (regular doses) (74.1 vs 28.6% of GP) (p < 0.05). Experience with omalizumab was reported only by 3.5%, of physicians, and higher rates among who were familiar with the guideline. CONCLUSION: This study shows that the knowledge of guideline recommendations in physicians who treat urticaria patients in Ecuador is low. The diagnostic workup and treatment of CsU patients are largely not in line with guideline recommendations in real life practice settings. We were able to compare results between German and Ecuadorian physicians and found that Ecuadorian physicians have lower awareness of the current guideline (33 vs 18%). Only one-third of physicians reported using regular doses of sgAHs as the first line treatment. Also, only 12.9% of physicians use sgAHs in higher doses and physicians still use fgAHs, particularly pediatricians (42.9%). Our results suggest that disparities in knowledge between physicians from different countries could influence the management of CsU. Knowledge of the guidelines is linked to better choices of treatments. Awareness of guidelines needs to be promoted for better management of chronic urticaria.

Exogenous lipid pneumonia related to long-term use of Vicks VapoRub® by an adult patient: a case report.

BACKGROUND: Use of petroleum-based over the counter remedies such as Vicks VapoRub to alleviate symptoms of rhinitis is common and can be effective, but carries under-appreciated risks of adverse side effects. In this case report we highlight Exogenous Lipoid Pneumonia (ELP), an uncommon condition that results from accumulation of exogenous lipids in the alveoli, as an adverse side effect of long-term Vicks VapoRub use. CASE PRESENTATION: We present the case of an 85-year-old female patient with ELP apparently due to continuous application of Vicks VapoRub® to her nostrils to alleviate chronic rhinitis. She was diagnosed incidentally via chest radiograph and computed tomography (CT) scan done as follow up to finding elevated C-reactive Protein during a routine exam. The CT scan revealed a pulmonary consolidation in the lower lobe of the right lung with fat density combined with low density areas associated with focal ground-glass opacities. The patient was advised to discontinue use of petroleum-based products, and was prescribed intranasal corticosteroids for her rhinitis. Follow up 2 years later showed that the lipid consolidation had diminished in size by approximately 10 %. CONCLUSION: Physicians must be aware that ELP can develop as a result of long-term application of petroleum-based oils and ointments to the nose and discourage such use of these products. Patients who have used petroleum-based products in this way should be screened for ELP. CT scan is the best imaging modality for establishing the diagnosis. The treatment of this condition is not well defined, but, as shown in this case, the size of the lipid mass can decrease after use of petroleum based substances is discontinued.

Characterization, cloning, and expression of two diagnostic antigens for Taenia solium tapeworm infection.

Adult and larval stages of Taenia solium cause 2 diseases in humans, i.e., taeniasis and cysticercosis, respectively. Diagnosis and treatment of taeniasis are the ultimate means to eliminate cysticercosis. A serological taeniasis diagnostic test has been developed for laboratory use. However, recombinant forms of the taeniasis diagnostic proteins are required to overcome the limited supply of native proteins and allow the development of a low-cost and field-applicable test with high sensitivity and specificity. Using 2-dimensional electrophoresis of T. solium excretory and secretory (TSES) products from hamster adult tape-worm in vitro cultures, we have identified 5 T. solium-specific protein spots, with molecular weights of 33 kDa (protein isoelectrofocusing point [pI]: 5.6, 5.3, 5.1) and 38 kDa (pI: 4.6, 4.5). Protein sequencing and molecular cloning of these proteins showed that although endowed with different pls, the proteins with the same molecular weights shared the same protein backbone, named TSES33 and TSES38. Their full-length complementary DNAs encode proteins with 267 and 278 amino acids, respectively. TSES33 and TSES38 were expressed in a baculovirus system. Both recombinant proteins were recognized by a panel of taeniasis, but not cysticercosis patient serum samples, indicating that they can potentially replace the native proteins in the development of a more efficacious taeniasis diagnostic test.