Identification and Characterization of Epithelial Cell-Derived Dense Bodies Produced upon Cytomegalovirus Infection

Dense bodies (DB) are complex, noninfectious particles produced during CMVinfection containing envelope and tegument proteins that may be ideal candidates as vaccines. Although DB were previously described in fibroblasts, no evidence of DB formation has been shown after propagating CMV in epithelial cells. In the present study, both fibroblast MRC-5 and epithelial ARPE-19 cells were used to study DB production during CMV infection. We demonstrate the formation of epithelial cell-derived DB, mostly located as cytoplasmic inclusions in the perinuclear area of the infected cell. DB were gradient-purified, and the nature of the viral particles was confirmed using CMV-specific immunelabeling. Epithelial cell-derived DB had higher density and more homogeneous size (200-300 nm) compared to fibroblast-derived DB (100-600 nm).In agreement with previous results characterizing DB from CMV-infected fibroblasts, the pp65 tegument protein was predominant in the epithelial cell-derived DB. Our results also suggest that epithelial cells had more CMV capsids in the cytoplasm and had spherical bodies compatible with nucleus condensation (pyknosis) in cells undergoing apoptosis that were not detected in MRC-5 infected cells at the tested time post-infection. Our results demonstrate the formation of DB in CMV-infected ARPE-19 epithelial cells that may be suitable candidate to develop a multiprotein vaccine with antigenic properties similar to that of the virions while not including the viral genome.This study was supported by the Spanish Ministry of Science, Innovation and University, Instituto de Salud Carlos III Grant/Award Numbers: PI17CIII-00014 (MPY110/18); PI20CIII-00009 (MPY303/20); DTS18CIII/00006 (MPY127/19). E.G-R is supported by the Sara Borrell Program (CD18CIII/00007), Instituto de Salud Carlos III, Ministerio de Ciencia, Innovación y Universidades. MJR is supported by the PTA Program (PTA2017-14233-I), Ministerio de Ciencia, Innovación y Universidades.S

A small non‐coding rna modulates expression of pilus‐1 type in streptococcus pneumoniae

Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide, and about 30% of the pneumococcal clinical isolates show type I pili‐like structures. These long protein-aceous polymers extending from the bacterial surface are encoded by pilus islet 1 and play major roles in adhesion and host colonization. Pili expression is bistable and is controlled by the transcriptional activator RlrA. In this work, we demonstrate that the previously identified small noncoding RNA srn135 also participates in pilus regulation. Our findings show that srn135 is generated upon processing of the 5′‐UTR region of rrgA messenger and its deletion prevents the synthesis of RrgA, the main pili adhesin. Moreover, overexpression of srn135 increases the expression of all pili genes and rises the percentage of piliated bacteria within a clonal population. This regulation is mediated by the stabilization of rlrA mRNA since higher levels of srn135 increase its half‐life to 165%. Our findings suggest that srn135 has a dual role in pilus expression acting both in cis‐ (on the RrgA levels) and in trans‐ (modulating the levels of RlrA) and contributes to the delicate balance between pili expressing and non‐expressing bacteria

Lippia origanoides Essential Oil or Thymol in Combination with Fluconazole Produces Damage to Cells and Reverses the Azole-Resistant Phenotype of a Candida tropicalis Strain

Candida tropicalis is one of the most pathogenic species within the genus. Increased antifungal resistance has been reported, which is in part due to the organism’s ability to form biofilms. In natural products derived from plants, such as essential oils (EOs) or their major components, there is significant potential to develop new antifungals or to both enhance the efficacy and reduce the toxicity of conventional antifungals. This study aimed to evaluate the effect of combining an EO of Lippia origanoides or thymol with fluconazole on an azole-resistant C. tropicalis strain. Synergism was observed in the combination of fluconazole with the EO and with thymol, and minimal inhibitory concentrations for fluconazole decreased at least 32-fold. As a consequence of the synergistic interactions, mitochondrial membrane potential was reduced, and mitochondrial superoxide production increased. Alteration in nuclear morphology, cell surface, and ultrastructure was also observed. In conclusion, the synergistic interaction between L. origanoides EO or thymol with fluconazole reverted the azole-resistant C. tropicalis phenotype. These findings suggest that L. origanoides EO or thymol alone, or in combination with fluconazole, have the potential for development as antifungal therapies for this yeast, including resistant strains

Four-Dimensional Characterization of the Babesia divergens Asexual Life Cycle, from the Trophozoite to the Multiparasite Stage

Babesia is an apicomplexan parasite of significance that causes the disease known as babesiosis in domestic and wild animals and in humans worldwide. Babesia infects vertebrate hosts and reproduces asexually by a form of binary fission within erythrocytes/red blood cells (RBCs), yielding a complex pleomorphic population of intraerythrocytic parasites. Seven of them, clearly visible in human RBCs infected with Babesia divergens, are considered the main forms and named single, double, and quadruple trophozoites, paired and double paired pyriforms, tetrad or Maltese Cross, and multiparasite stage. However, these main intraerythrocytic forms coexist with RBCs infected with transient parasite combinations of unclear origin and development. In fact, little is understood about how Babesia builds this complex population during its asexual life cycle. By combining cryo-soft X-ray tomography and video microscopy, main and transitory parasites were characterized in a native whole cellular context and at nanometric resolution. The architecture and kinetics of the parasite population was observed in detail and provide additional data to the previous B. divergens asexual life cycle model that was built on light microscopy. Importantly, the process of multiplication by binary fission, involving budding, was visualized in live parasites for the first time, revealing that fundamental changes in cell shape and continuous rounds of multiplication occur as the parasites go through their asexual multiplication cycle. A four-dimensional asexual life cycle model was built highlighting the origin of several transient morphological forms that, surprisingly, intersperse in a chronological order between one main stage and the next in the cycle. IMPORTANCE Babesiosis is a disease caused by intraerythrocytic Babesia parasites, which possess many clinical features that are similar to those of malaria. This worldwide disease is increasing in frequency and geographical range and has a significant impact on human and animal health. Babesia divergens is one of the species responsible for human and cattle babesiosis causing death unless treated promptly. When B. divergens infects its vertebrate hosts, it reproduces asexually within red blood cells. During its asexual life cycle, B. divergens builds a population of numerous in-traerythrocytic (IE) parasites of difficult interpretation. This complex population is largely unexplored, and we have therefore combined three- and four-dimensional imaging techniques to elucidate the origin, architecture, and kinetics of IE parasites. Unveiling the nature of these parasites has provided a vision of the B. divergens asexual cycle in unprecedented detail and is a key step to develop control strategies against babesiosis

Developing a Critical Understanding of Smart Urbanism?

Smart urbanism is emerging at the intersection of visions for the future of urban places, new technologies and infrastructures. Smart urbanism discourses are deeply rooted in seductive and normative visions of the future where digital technology stands as the primary driver for change. Yet our understanding of the opportunities, challenges, and implications of smart urbanism is limited. Research in this field is in its infancy, fragmented along disciplinary lines and often based on single city case studies. As a result, we lack both the theoretical insight and empirical evidence required to assess the implications of this potentially transformative phenomenon. Given the significant implications of smart urbanism there is an urgent need to critically engage with why, how, for whom and with what consequences smart urbanism is emerging in different urban contexts. The aim of this review is to unpack the different logics and rationales behind smart urbanism discourses and proposals, and in this way understand the ways by which imaginaries of urban futures are currently being constructed, along with their socio-technical and political implications for future research priorities

MicroRNA signature from extracellular vesicles of HCV/HIV co-infected individuals differs from HCV mono-infected

Hepatitis C virus (HCV) coinfection with human immunodeficiency virus (HIV) has a detrimental impact on disease progression. Increasing evidence points to extracellular vesicles (EVs) as important players of the host-viral cross-talk. The microRNAs (miRNAs), as essential components of EVs cargo, are key regulators of normal cellular processes and also promote viral replication, viral pathogenesis, and disease progression. We aimed to characterize the plasma-derived EVs miRNA signature of chronic HCV infected and HIV coinfected patients to unravel the molecular mechanisms of coinfection. EVs were purified and characterized from 50 plasma samples (21 HCV mono- and 29 HCV/HIV co-infected). EV-derived small RNAs were isolated and analyzed by massive sequencing. Known and de novo miRNAs were identified with miRDeep2. Significant differentially expressed (SDE) miRNA identification was performed with generalized linear models and their putative dysregulated biological pathways were evaluated. Study groups were similar for most clinical and epidemiological characteristics. No differences were observed in EVs size or concentration between groups. Therefore, HCV/HIV co-infection condition did not affect the concentration or size of EVs but produced a disturbance in plasma-derived EVs miRNA cargo. Thus, a total of 149 miRNAs were identified (143 known and 6 de novo) leading to 37 SDE miRNAs of which 15 were upregulated and 22 downregulated in HCV/HIV co-infected patients. SDE miRNAs regulate genes involved in inflammation, fibrosis, and cancer, modulating different biological pathways related to HCV and HIV pathogenesis. These findings may help to develop new generation biomarkers and treatment strategies, in addition to elucidate the mechanisms underlying virus-host interaction. KEY MESSAGES: HCV and HCV/HIV displayed similar plasma-EV size and concentration. EVs- derived miRNA profile was characterized by NGS. 37 SDE miRNAs between HCV and HCV/HIV were observed. SDE miRNAs regulate genes involved in inflammation, fibrosis and cancer.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work has been supported by grants from (1) Institute of Health Carlos III, Spain [PI18CIII/00020/ to AFR], (2) PID2021–126781OB-I00 funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”, (3) The SPANISH AIDS Research Network RD16CIII/0002/0002 - ISCIII – FEDER, (4) Centro de Investigación en Red en Enfermedades Infecciosas (CIBERINFEC) CB21/13/00044, (5) the National Agency for Scientific and Technology Promotion (ANPCyT) (PICT 2017 Nº713), and (6) the National Research Council (CONICET, PIP 2021-2023). V.C. received funding form the Asociación Universitaria Iberoamericana de Postgrado (AUIP) for the Academic Mobility Scholarship Program. P.V., E.D.M., and M.V.P. are members of the CONICET-Research Career Program. V.C. is a fellow from ANPCyT. The funder’s had no role in the study design, data collection and analysis, decision to publish, or the preparation of the manuscript.S

Diagnósticos enfermeros en UFISS, UGA, Traumatología y CIR.

In 2006, after the addition of a new nurse in UFISS (Social-Sanitary Functional Interdisciplinary Unit), is detected the need of a common language for all nurses with which to conduct a data collection for the nursing reports. The aim of this study is to know the main nursing diagnoses in UFISS, Geriatrics, Traumatology and Surgery units, using the NANDA (North American Nursing Diagnosis Association) nursing diagnostic terminology. We performed a retrospective study of all the medical reports at UFISS in 2006, a prospective study of all medical reports at Geriatrics and geriatric patients at Traumatology and Surgery in various periods between 2008 and 2009. According to the results, the use of a validated method like NANDA diagnoses, has enabled to nurses to identify common altered needs and after that, to define Nursing Diagnoses and develop the optimal care plan for the patientIntroducción: La utilización de un método validado, ha permitido detectar las necesidades alteradas en relación a su etiología, definiendo los DdE (Diagnósticos de Enfermería) (3).La utilización de la Taxonomía NANDA 2 asegura la definición de la respuesta humana a un problema tanto dentro del marco profesional como jurídico, así mismo permite un lenguaje común en la práctica enfermera (6).Objetivos. Identificar los DdE más prevalentes en la población atendida por la UFISS (Unidad Funcional Interdisciplinar Socio-Sanitaria), UGA (Unidad de Geriatría Aguda), TRAUMATOLOGÍA y CIRUGÍA de la FHAG (Fundación Hospital Asilo de Granollers) utilizando la taxonomía NANDA.Métodos.UFFIS. Se estudian retrospectivamente todas las historias de la UFISS del año 2006. (674 consultas, entrando en estudio N= 390 estudiadas).Se estudian prospectivamente:COT (Cirugía Ortopédica Traumatológica). 24 pacientes geriátricos de la unidad de trauma ingresados durante los meses de Agosto-Septiembre 2008UGA. 49 pacientes ingresados durante los meses de Septiembre-Diciembre 2008CIR. 36 pacientes ingresados en mayo 2009.Valoración paciente: Abordaje Bio-Psico-Social (Entrevista enfermera al paciente y al cuidador principal).Funcional (Barthel). Instrumentales (Lawton). Cognitivo (Pfeiffer). Riesgo de úlceras (EMINA). Dolor (EVA)Resultados.UFISS: Se detectan 18 diagnósticos, como los más prevalentes valorados en la UFISS,COT: Se detectan 26 diagnósticos, 16 son comunes a los recogidos por la enfermera de la UFISS, los 10 restantes son los específicos detectados en el paciente orto geriátrico:UGA: Se detectan 30 diagnósticos, de los cuales 18 son comunes a la UFISS y los 12 restantes son específicos en el paciente geriátrico.CIR: Se detectan alrededor de 50 diagnósticos; pendiente tabulación final.Conclusiones: Se han definido los diagnósticos más prevalentes determinando los comunes a las diferentes áreas asistenciales. Dado que en nuestra institución la formación es mayoritariamente básica, con este estudio hemos conseguido: 1) difundir el lenguaje NANDA, 2) asegurar el dominio de estos diagnósticos, 3) que el profesional trabaje de forma más segura utilizando un lenguaje validado y entendible y 4) orientar a la futura implantación informática

Metabolic Impact of Adult-Onset, Isolated, Growth Hormone Deficiency (AOiGHD) Due to Destruction of Pituitary Somatotropes

Growth hormone (GH) inhibits fat accumulation and promotes protein accretion, therefore the fall in GH observed with weight gain and normal aging may contribute to metabolic dysfunction. To directly test this hypothesis a novel mouse model of adult onset-isolated GH deficiency (AOiGHD) was generated by cross breeding rat GH promoter-driven Cre recombinase mice (Cre) with inducible diphtheria toxin receptor mice (iDTR) and treating adult Cre+/−,iDTR+/− offspring with DT to selectively destroy the somatotrope population of the anterior pituitary gland, leading to a reduction in circulating GH and IGF-I levels. DT-treated Cre−/−,iDTR+/− mice were used as GH-intact controls. AOiGHD improved whole body insulin sensitivity in both low-fat and high-fat fed mice. Consistent with improved insulin sensitivity, indirect calorimetry revealed AOiGHD mice preferentially utilized carbohydrates for energy metabolism, as compared to GH-intact controls. In high-fat, but not low-fat fed AOiGHD mice, fat mass increased, hepatic lipids decreased and glucose clearance and insulin output were impaired. These results suggest the age-related decline in GH helps to preserve systemic insulin sensitivity, and in the context of moderate caloric intake, prevents the deterioration in metabolic function. However, in the context of excess caloric intake, low GH leads to impaired insulin output, and thereby could contribute to the development of diabetes

Epigenetic and post-transcriptional regulation of somatostatin receptor subtype 5 (SST5 ) in pituitary and pancreatic neuroendocrine tumors.

Somatostatin receptor subtype 5 (SST5 ) is an emerging biomarker and actionable target in pituitary (PitNETs) and pancreatic (PanNETs) neuroendocrine tumors. Transcriptional and epigenetic regulation of SSTR5 gene expression and mRNA biogenesis is poorly understood. Recently, an overlapping natural antisense transcript, SSTR5-AS1, potentially regulating SSTR5 expression, was identified. We aimed to elucidate whether epigenetic processes contribute to the regulation of SSTR5 expression in PitNETs (somatotropinomas) and PanNETs. We analyzed the SSTR5/SSTR5-AS1 human locus in silico to identify CpG islands. SSTR5 and SSTR5-AS1 expression was assessed by quantitative real-time PCR (qPCR) in 27 somatotropinomas, 11 normal pituitaries (NPs), and 15 PanNETs/paired adjacent (control) samples. We evaluated methylation grade in four CpG islands in the SSTR5/SSTR5-AS1 genes. Results revealed that SSTR5 and SSTR5-AS1 were directly correlated in NP, somatotropinoma and PanNET samples. Interestingly, selected CpG islands were differentially methylated in somatotropinomas compared with NPs. In PanNETs cell lines, SSTR5-AS1 silencing downregulated SSTR5 expression, altered aggressiveness features, and influenced pasireotide response. These results provide evidence that SSTR5 expression in PitNETs and PanNETs can be epigenetically regulated by the SSTR5-AS1 antisense transcript and, indirectly, by DNA methylation, which may thereby impact tumor behavior and treatment response

Isoetes pedersenii, a new species from Southern South America

The name Isoetes pedersenii H.P. Fuchs (Lycophyta), a species known only from the Mburucuyá National Park, Corrientes, Argentina, is validated. Observations were carried out on herbarium material with stereoscopic, light and scanning electron microscopes. The species is described and typified. A diagnosis and discussion about its distribution and its relationship with the morphology of other species of Isoetes are provided.O nome Isoetes pedersenii H.P. Fuchs (Lycophyta) foi validado para a espécie identificada apenas no Parque Nacional de Mburucuyá, em Corrientes na Argentina. Material preservado em herbário foi avaliado com microscópios estereoscó pico, de luz branca e eletrônico de varredura. A espécie foi descrita e tipificada. Um diagnóstico e uma discussão sobre a distribuição e relação com a morfologia de outras espécies de Isoetes são relatados