Active immunization with a structurally aggregated PD-L1 antigen breaks T and B immune tolerance in non-human primates and exhibits in vivo anti-tumoral effects in immunocompetent mouse tumor models.

Despite the clinical success of the programmed death ligand 1 (PD-L1) blocking therapy in cancer treatment, only a subset of patients exhibits durable responses, therefore further exploration of other immunotherapeutic alternatives are needed. This paper reported the development of the PKPD-L1Vac vaccine, a new protein vaccine candidate that uses aluminum phosphate as an adjuvant and as an antigen the extracellular domain of human PD-L1 fused to a 47 amino-terminal portion of the LpdA protein from N. meningitides (PKPD-L1). The PKPD-L1 antigen has different physical and biological characteristics than those found in the natural molecule and in others PD-L1 vaccine candidates. The quimeric protein has a reduced binding capacity to the PD-1 and CD80 receptors to decrease their pro-tumoral activity. Besides, the distinctive feature of the PKPD-L1 polypeptide to be structurally aggregated could be desirable for its immunogenic properties. PKPD-L1Vac elicited anti-PD-L1-specific IgG antibodies and T lymphocyte-mediated immunity in mice and non-human primates. The vaccine administration demonstrated antitumor activity on CT-26 and B16-F10 primary tumor models in mice. Moreover, the immunization with PKPD-L1Vac increased the tumor-infiltrating lymphocytes and decreased the proportion of CD3+CD8+PD1+high anergic T cells in CT-26 tumor tissues, suggesting that the vaccine may remodel the tumor microenvironment. In summary, the PKPD-L1Vac vaccine exhibits very promising preclinical results and deserves to move forward to a phase I clinical trial.

Utilidad de los parámetros clínicos y de laboratorio básicos para predecir infección bacteriana seria en menores de 3 meses que se hospitalizan por síndrome febril sin foco / Utility of basic clinical and laboratory parameters to predict Serious Bacterial Infection in children younger than 3 months old hospitalized for Febrile Syndrome without Source

Resumen: Introducción: Un 20% de los niños con síndrome febril se presenta como síndrome febril sin foco (SFSF). Las es trategias de manejo en este grupo presentan alta sensibilidad, pero baja especificidad. Objetivos: Ca racterizar las infecciones bacterianas serias (IBS) en menores de 3 meses hospitalizados por SFSF, y evaluar utilidad de parámetros clínicos y de laboratorio en la identificación de pacientes con alto riesgo de IBS. Pacientes y Método: Estudio prospectivo en pacientes < 3 meses hospitalizados entre enero 2014 y noviembre 2015 por SFSF en dos hospitales pediátricos de la Región Metropolitana. Criterios de inclusión: edad 4 días - 3 meses, fiebre > 38°C de < 72 h de evolución sin causa demostra ble. Criterios de exclusión: uso de antimicrobianos hasta 7 días previo a su ingreso, prematuros < 34 semanas, peso de nacimiento < 2 kg e inmunocomprometidos. Se registraron datos demográficos, clínicos, y exámenes de laboratorio, hemograma y PCR, diagnóstico de egreso, IBS descartada, IBS probable o confirmada. Resultados: 32% de los pacientes egresó con diagnóstico de IBS, 28% con diagnóstico de infección viral o probablemente viral, 34% con diagnóstico de SFSF no especificado y 6% SFSF por otras causas. No se encontraron diferencias significativas en PCR, leucocitosis, aspecto tóxico ni horas de fiebre al ingreso al comparar los grupos con y sin IBS (p > 0,05). La combinación de parámetros clínicos y de laboratorio mostro sensibilidad de 27%, especificidad de 90%, VPP 60% y VPN 71%. Conclusión: No fue posible establecer que parámetros clínicos y de laboratorio permitan identificar menores de 3 meses con alto riesgo de IBS, manteniendo su utilidad como indicadores de bajo riesgo. Es necesario contar con otros elementos clínicos y de laboratorio que permitan discrimi nar IBS de infecciones virales.

Abstract: Introduction: In 20% of children with febrile syndrome, it appears as fever of unknown origin (FUO) syndrome. Management strategies in this group have high sensitivity but low specificity. Objectives: To cha racterize serious bacterial infections (SBI) in children younger than three months old hospitalized because of FUO syndrome and to evaluate the utility of clinical and laboratory parameters in the identification of patients that are at high risk of SBI. Patients and Method: Prospective study in patients aged < 3 months hospitalized due to FUO syndrome between January 2014 and November 2015 in two pediatric hospitals in the Metropolitan Region. Inclusion criteria: age 4 days - 3 months, fever > 38°C longer than 72 hours after onset without demonstrable cause. Exclusion criteria: anti microbial use up to 7 days before admission, preterm infants < 34 weeks, birth weight < 2 kg, and im munocompromised. Demographic, clinical, and laboratory tests data were recorded as well as blood count and CRP, discharge diagnosis, and ruled out, probable or confirmed SBI. Results: 32% of the patients were discharged with diagnosis of SBI, 28% with diagnosis of viral or probably viral infec tion, 34% with diagnosis of not specified FUO syndrome, and 6% due to other causes. There were no significant differences in the CRP value, altered WBCs count, toxic aspect, or hours of fever at the admission when comparing groups with and without SBI (p < 0.05). The combination of clinical and laboratory parameters showed 27% of sensitivity, 90% of specificity, 60% of PPV, and 71% of NPV. Conclusion: It was not possible to establish clinical and laboratory parameters that allow the identifi cation of children younger than 3 months old at high risk of SBI, however, they maintain their value as low risk indicators. It is necessary further investigation of other clinical and laboratory elements that allow discriminating SBI from viral infections.

The impact of time from biopsy to radical prostatectomy on Gleason score undergrading and other related factors. / Impacto del tiempo desde la biopsia a la prostatectomía radical en la infragradación del grado de Gleason y análisis de factores relacionados.

INTRODUCTION: Gleason score biopsy undergrading (GSBU) can have an impact on the management and prognosis of patients with prostate cancer. We analyze the possible impact of time and other clinical and analytical factors in the appearance of GSBU in our series. PATIENTS AND METHOD: Ambispective, multicenter study of 1955 patients with localized prostate cancer undergoing radical prostatectomy between 2005 and 2018. Descriptive statistics and hypothesis testing are reported by univariate and multivariate analyses. RESULTS: Mean age 63.69 (44-80) years, median PSA 8.70 ng / ml (1.23-99). GSBU was observed in 34.7% of the entire cohort. In 72.8% of the cases, the GSBU occurred in one consecutive Gleason score, with the progression from 3 + 3 to 3 + 4 being the most frequent (289 patients, 47.6%). Performing radical prostatectomy 90-180 days before or after the biopsy does not have an impact on its undergrading in any of the groups. In the univariate and multivariate analysis, the presence of tumor or pathological rectal examination in both lobes, the tumor load ≥50% of cylinders and a DPSA ≥0.20, showed independent discriminative capacity to select patients who presented GSBU. CONCLUSIONS: The time from biopsy to radical prostatectomy did not show impact on GSBU. The number of affected cylinders, bilateral tumor and DPSA are easily accessible parameters that can help us select patients with greater probability of presenting GSBU.

Actual incidence of prostate cancer in healthcare areas of the autonomous community of Castilla-Leon during 2014. CAPCYL registry data. / Incidencia real de cáncer de próstata en las áreas sanitarias de la comunidad autónoma de Castilla y León durante el año 2014. Datos del registro CAPCYL.

OBJECTIVE: To determine the actual incidence of prostate cancer (PC) in the healthcare areas of Castilla-Leon in 2014. MATERIAL AND METHODS: A multicentre study was conducted with the participation of 7 of the 9 healthcare areas of Castilla-Leon. We collected retrospective data that included 87.8% of the target population (men diagnosed with PC with histopathological confirmation in 2014). We calculated the raw and age-adjusted incidence rates based on the direct method and consulted the community and national epidemiological data in the Spanish National Institute of Statistics. RESULTS: A total of 1198 new cases of PC were diagnosed, with a raw incidence rate in the community of 109.54 cases per 100,000 men. The adjusted rates for the Spanish and European populations were 115.41 and 110.07, respectively. The age group with the highest diagnostic concentration was the 60-70-year group, with 41.97% of the diagnoses. The group with the highest incidence was the 70-80-year group, with 438.87 cases per 100,000 inhabitants. There were differences in the raw and age-adjusted incidence rates and in the age at diagnosis among the various included healthcare areas. CONCLUSIONS: The community raw incidence rate was higher than most existing data. We observed significant differences among the various geographical areas, which could be explained mainly by the age distribution and the opportunistic screening policies for each area.

Functionality based method for simultaneous isolation of rodent hepatic sinusoidal cells.

Chronic liver disease is the result of long term exposure to viruses or toxins such as alcohol, fat and drugs, and forms the basis for the development of liver fibrosis and primary liver cancer. In vitro and in vivo models are key to study the pathways involved in chronic liver disease and for the development of therapeutics. 3D co-culture systems are becoming the in vitro standard, which requires freshly isolated primary hepatic cells. We developed a novel isolation method to simultaneously isolate liver sinusoidal endothelial cells (LSECs), Kupffer cells (KCs) and hepatic stellate cells (HSCs). The method exploits the scavenging activity of LSECs, the phagocytic capacity of KCs and the retinoid content of HSCs in vivo to enable direct processing by fluorescence-activated cell sorting without additional antibody binding and washing steps. UFACS3, for UV-FACS-based isolation of 3 non-parenchymal liver cell types, yields functional and pure LSECs (98 ± 1%), KCs (98 ± 1%) and HSCs (97 ± 3%), with less hands-on time from healthy and diseased rodent livers. This novel approach allows a fast and effective combined isolation of sinusoidal cells for further analysis.

Factibilidad de la interpretación de espirometrías en preescolares según criterios ATS/ERS / Spirometry interpretation feasibility among pre-school children according to the European Respiratory Society and American Thoracic Society Guidelines

Introducción: La espirometría es el examen más utilizado para evaluar la función pulmonar. El año 2007 se publicaron guías que definieron criterios de aceptabilidad y repetibilidad para su realización e interpretación en preescolares. Nuestro objetivo fue describir las espirometrías de pacientes de este grupo etario según el cumplimiento de estos criterios. Pacientes y Método: Se revisaron las espirometrías basales de pacientes de 2 a 5 años realizadas en el Laboratorio de Función Pulmonar Pediátrico de la P. Universidad Católica de Chile derivados por tos o sibilancias recurrentes o persistentes. Se consideraron solo las obtenidas en pacientes que la realizaban por primera vez. Se analizaron según criterios internacionales. Resultados: Se obtuvieron 93 espirometrías (edad promedio 57,4 ± 8,6 meses, 48 varones): 44 (47%) tuvieron todos los criterios aceptables, 87 (93%) obtuvieron un tiempo espiratorio ≥ 0,5 segundos, 67 (72%) de los pacientes tuvieron un flujo espiratorio de final de espiración en valor ≤ 10% del flujo espiratorio máximo. La variabilidad de las mediciones de capacidad vital forzada (CVF) y volumen espirado al primer segundo (VEF1) fue muy baja (coeficiente de correlación intraclase > 0,9). Conclusión: En nuestro centro fue factible cumplir criterios de aceptabilidad y repetibilidad en espirometrías en preescolares, semejante a descripciones previas. Al igual que en niños mayores, se recomienda realizar este examen en preescolares que requieren estudio de la función pulmonar.

Introduction: Spirometry is the most used test to evaluate pulmonary function. Guidelines that defined acceptability and repeatability criteria for its implementation and interpretation among preschoolers were published in 2007. Our objective was to quantify the actual compliance with these criteria among pre-school patients. Methods: A review was performed on the baseline spirometry measured in patients aged 2 to 5 years in the Pediatric Respiratory Laboratory of the Pontificia Universidad Catolica de Chile, who were admitted due to recurrent or persistent coughing or wheezing. Only those results obtained in patients who took the test for the first time were considered. They were analyzed by international standards. Results: A total of 93 spirometry results (mean age 57.4 ± 8.6 months, 48 males) were obtained, of which 44 (47%) met all acceptable criteria, 87 (93%) obtained expiratory time of ≥ 0.5 seconds, and 67 (72 %) of the patients had an end-expiratory flow of ≤ 10% from peak flow. The variation in the measurement of forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) was very low (intraclass correlation coefficient > 0.9). Conclusion: It was possible to meet the acceptability and repeatability criteria for spirometry among pre-school children in our Center, which was similar to previous reports. As in older children, this test is fully recommended for pre-school children who require lung function studies.

Heart pigmentation in the gray bichir, Polypterus senegalus (Actinopterygii: Polypteriformes).

The occurrence of pigment cells in the heart is well documented in amphibians, birds and mammals. By contrast, information on heart pigmentation in fish is extremely sparse. The aim is to report the presence of pigment cells over the entire surface of the heart in the gray bichir, Polypterus senegalus. The sample consisted of 12 hearts, which, after gross anatomical examination, were studied using histochemical and immunohistochemical techniques for light microscopy, and transmission electron microscopy. The pigment cells were located in the subepicardium, showing a regular distribution pattern across the whole heart, except for the anterior end of the outflow tract, where the pigmentation was much more intense. The cells contained dark, ovoid-shaped organelles which was consistent with a melanosome cell identity. As in other vertebrates, the physiological role of the pigment cells in the heart of the gray bichir is unknown. The absence of such cells in hearts of other polypteriforms suggests that cells containing melanin are not essential for normal fish heart function. Basing on literature data concerning tetrapods, it can be inferred that the pigment cells of the heart of the gray bichir derive from the neural crest. If this were true, our findings would provide the first evidence for the presence of neural crest-derived cells in the subepicardium of adult hearts of early actinopterygians.

[Perioperative treatment for the urgent orbital decompression surgery in a 30-weeks pregnant woman with Graves' orbitopathy]. / Tratamiento perioperatorio para la cirugía urgente de descompresión orbitaria en una paciente gestante de 30 semanas con orbitopatía de Graves.

Thyroid ophthalmopathy is a rare extra-thyroid complication usually associated with Graves' disease. This disease can occur in the euthyroid pregnant patient. Graves' orbitopathy is characterized by eyelid retraction, proptosis, extraocular muscle dysfunction, and periorbital edema. In some cases an emergency surgical repair may be required to avoid irreversible vision loss. We present the case of a 35-year-old woman in her 30th gestational week, who suffered from Graves' ophthalmopathy, severe exophthalmia, and visual acuity decrease. Following consultations among anesthesiologists, ophthalmologists, maxillofacial surgeons, endocrinologists, obstetricians and the patient, it was decided to perform a surgical orbital wall decompression. The anesthetic and perioperative implications associated with gestational age and the considerations for this surgical procedure, and how to avoid increasing intraocular pressure are discussed.

Syncope.

Syncope, which can be defined as a transient loss of consciousness caused by transient global cerebral hypoperfusion and characterised by rapid onset, short duration, and spontaneous complete recovery, is a common condition. This definition is useful for differentiating syncope from other clinical conditions that also involve real or apparent transient loss of consciousness, but in which the mechanism is not global cerebral hypoperfusion, such as epilepsy, falls, or psychiatric pseudosyncope. We reviewed the etiological classification of syncope and found that reflex syncope is the most common etiology in the general population, with a good prognosis, whereas cardiac syncope increases with age and has a worse prognosis. We also reviewed the role and limitations of different tests, specifically referring to the interpretation of the results of carotid sinus massage, the role of tilt-table testing, the diagnostic strategy in patients with syncope and bundle branch block, the adenosine test, and the emerging role of prolonged electrocardiographic monitoring. Furthermore, we reviewed the different therapeutic options available. The importance of establishing syncope units with the aim of improving the diagnostic process and optimizing resources is also emphasized.

Relationships between optically assessed polyphenols and chlorophyll contents, and leaf mass per area ratio in woody plants: a signature of the carbon-nitrogen balance within leaves?

Chlorophyll (Chl) and epidermal polyphenol (EPhen) contents were estimated in vivo using two optical leaf-clips, SPAD-502 and Dualex, respectively. The area-based measurements were transformed into mass-based data by taking into account the leaf dry mass per area (LMA). Measurements were performed on forest trees and on saplings grown under controlled conditions. While LMA increased with irradiance along a vertical transect in a beech canopy or in saplings grown under different and increasing irradiance levels, mass-based EPhen (EPhen(m)) increased, whereas mass-based Chl (Chl(m)) decreased. This was a signature of a gradual switch of investment from protein into polyphenol production. A similar signature was obtained in saplings grown on nitrogen-deficient soil with respect to fertilized controls. However, nitrogen effects remained moderate compared to irradiance-induced effects. EPhen(m) and Chl(m) both declined with plant ageing-induced increases in LMA, under all tested growth conditions. This was a signature of an accumulation of dry matter that diluted Chl and EPhen. The described competition between Chl and EPhen in leaves fits well with the predictions of the Protein Competition Model (PCM), that is, that the total leaf mass-based polyphenols content (Phen(t)) is controlled by the competition between protein and polyphenol biosynthetic pathways and its metabolic regulation.

Iron deficiency interrupts energy transfer from a disconnected part of the antenna to the rest of Photosystem II.

Iron deficiency changed markedly the shape of the leaf chlorophyll fluorescence induction kinetics during a dark-light transition, the so-called Kautsky effect. Changes in chlorophyll fluorescence lifetime and yield were observed, increasing largely the minimal and the intermediate chlorophyll fluorescence levels, with a marked dip between the intermediate and the maximum levels and loss of the secondary peak after the maximum. During the slow changes, the lifetime-yield relationship was found to be linear and curvilinear (towards positive lifetime values) in control and Fe-deficient leaves, respectively. These results suggested that part of the Photosystem II antenna in Fe-deficient leaves emits fluorescence with a long lifetime. In dark-adapted Fe-deficient leaves, measurements in the picosecond-nanosecond time domain confirmed the presence of a 3.3-ns component, contributing to 15% of the total fluorescence. Computer simulations revealed that upon illumination such contribution is also present and remains constant, indicating that energy transfer is partially interrupted in Fe-deficient leaves. Photosystem II-enriched membrane fractions containing different pigment-protein complexes were isolated from control and Fe-deficient leaves and characterized spectrophotometrically. The photosynthetic pigment composition of the fractions was also determined. Data revealed the presence of a novel pigment-protein complex induced by Fe deficiency and an enrichment of internal relative to peripheral antenna complexes. The data suggest a partial disconnection between internal Photosystem II antenna complexes and the reaction center, which could lead to an underestimation of the Photosystem II efficiency in dark-adapted, low chlorophyll Fe-deficient leaves, using chlorophyll fluorescence.

[The tilt-table test: is it essential for adequate treatment of vasovagal syncope? Arguments in favor]. / Test de mesa basculante: es imprescindible para el tratamiento adecuado del síncope vasovagal? Argumentos a favor.

Head-Up tilt test (HUT) permits the identification of vasovagal syncope in patients with syncope of unknown origin. Several authors have pointed out its usefulness for the therapeutic management of these patients. However, even though it has limitations, this paper discusses those situations in which HUT may be useful. First, as HUT permits the recognition of the etiologic diagnosis of syncope, it may be helpful in the evaluation of its treatment. On the other hand, the characterization of different components of vasovagal reaction, cardioinhibition and/or vasodepressor response, may be helpful in the selection of therapeutic options. It has been suggested that the performance of HUT may even be beneficial for the relapses of vasovagal syncope in patients with recurrent syncope and refractory to other treatments. Finally, further randomized clinical trials in HUT will help to establish the prediction of therapeutic effectiveness in patients with vasovagal syncope. However, there are important limitations to its usefulness for the clinical management of individual patients.

Characterization of Blue-Green Fluorescence in the Mesophyll of Sugar Beet (Beta vulgaris L.) Leaves Affected by Iron Deficiency.

The mesophyll of sugar beet (Beta vulgaris L.) leaves emits red (chlorophyll a) fluorescence and blue-green fluorescence when excited with ultraviolet light. The intensity of blue-green fluorescence was increased in mesophylls affected by iron deficiency. This increase was large and progressive. It was concomitant with a decrease of photosynthetic pigments per unit of leaf area. Most of the increase in blue-green fluorescence can be explained by the decrease of the screening of ultraviolet light by chlorophylls and carotenoids. In addition, chlorophylls selectively reabsorb blue fluorescence, which leads to a change in the form of the fluorescence emission spectra. This effect induces an increase of the blue-to-green fluorescence ratio in control mesophylls that was concomitant with the decrease of chlorophyll per unit of leaf area. Iron deficiency induced a decrease of the blue-to-green fluorescence ratio that may be attributed to an accumulation of flavins fluorescing in the green. Time-resolved fluorescence measurements indicate that they are mostly riboflavin and/or flavin mononucleotide phosphate. Our data also indicate that the blue-green fluorescence emitted from the mesophyll contains fluorescence of nicotinamide nucleotides.

[Etiologic diagnosis of pneumonia in adults: usefulness of bronchoalveolar lavage]. / Diagnóstico etiológico de la neumonia del adulto: utilidad del lavado broncoalveolar.

Quantitative evaluation of bacteria in bronchoalveolar lavage fluid from 15 patients with pneumonia was compared to that in 29 healthy controls. Aerobic and anaerobic cultures were used as well as cultures and staining for fungi and mycobacteria. A total of 1000 colony forming units per ml was considered a cutoff mark between colonization and infection. A positive result was obtained in 13 of 15 patients with pneumonia, allowing the identification of the causing agent. Counts below the indicated level were observed in 23 of 29 controls. There was no morbidity associated to the procedure. Thus, an 85% sensitivity and 87% specificity for bronchoalveolar lavage in the etiologic diagnosis of pneumonia may be estimated from this study.

[Hypothyroid myopathy. Clinico-pathologic study of 20 cases]. / Miopatía hipotiroidea. Estudio clínico-patológico de 20 casos.

20 patients afflicted with primary hypothyroidism were studied in order to evaluate the association of clinical or sub-clinical myopathy, detected by neurophysiological (electromyography) (EMG) or neuropathological methods (muscular biopsy with enzymatic study). 70% of the patients had muscular weakness (moderate in 30% and severe in 40%) of the scapular and pelvic muscles. 60% of the patients had muscular cramps. There was no myodema nor muscular atrophy or hypertrophy. Seric CPK was high in 70% of the cases. EMG was myopathic in 65%. All cases with weakness registered EMG alterations. The histological findings were import findings were important. The enzymatic techniques showed alterations of the fiber subtypes in 90% of the cases. The type I fibers had sarcolemmal and mitochondrial accumules in 85% and 70% had areas without oxidative activity, similar to "core". In this study, we did not find any correlation between the evolution time of hypothyroidism, hormonal levels, CPK increase, and muscular weakness. The EMG was myopathic in cases with severe weakness, however, in patients with moderate weakness it could also prove abnormal. There was no correlation between the electric myopathic pattern, CPK levels and thyroid hormones.

[Thyroid myopathy. Effect of treatment with thyroid hormones]. / Miopatía tiroidea. Efecto del tratamiento con hormonas tiroideas.

We studied 20 patients afflicted with primary hypothyroidism and treated with thyroid hormone replacements, with the aim of evaluating the clinical, biochemical, EMG and anatomo-pathological effects. Cramps, as well as subjective and objective proximal muscular weakness, disappeared. Seric CPK was normalized in all cases after 1-8 weeks of treatment. EMG myopathic patterns faded away in all cases. Anatomopathological changes took longer to observe; 5 cases with new biopsy showed an isolate atrophy of type II fibers, one of them after 18 months in treatment; type I fibers had mitochondrial accumula without change compared to the initial biopsy. There was a clear tendency for the destroyed fibers to become normalized and for the structures similar to the "core" to vanish.