The nuclear import receptor Kapß2 modifies neurotoxicity mediated by poly(GR) in C9orf72-linked ALS/FTD.

Expanded intronic G4C2 repeats in the C9ORF72 gene cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). These intronic repeats are translated through a non-AUG-dependent mechanism into five different dipeptide repeat proteins (DPRs), including poly-glycine-arginine (GR), which is aggregation-prone and neurotoxic. Here, we report that Kapß2 and GR interact, co-aggregating, in cultured neurons in-vitro and CNS tissue in-vivo. Importantly, this interaction significantly decreased the risk of death of cultured GR-expressing neurons. Downregulation of Kapß2 is detrimental to their survival, whereas increased Kapß2 levels mitigated GR-mediated neurotoxicity. As expected, GR-expressing neurons displayed TDP-43 nuclear loss. Raising Kapß2 levels did not restore TDP-43 into the nucleus, nor did alter the dynamic properties of GR aggregates. Overall, our findings support the design of therapeutic strategies aimed at up-regulating Kapß2 expression levels as a potential new avenue for contrasting neurodegeneration in C9orf72-ALS/FTD.

Resultados funcionales de tratamiento con bevacizumab intravítreo en pacientes con retinopatía del prematuro / Functional results in children with retinopathy of prematurity treated with intravitreal bevacizumab

Objetivo El propósito del presente estudio es determinar la eficacia y describir los resultados funcionales en términos de agudeza visual y defecto refractivo a largo plazo del tratamiento con una dosis de bevacizumab intravítreo en pacientes con retinopatía del prematuro (ROP) tipo1 de alto riesgo. Métodos Se trata de un estudio clínico retrospectivo en el que se seleccionaron todos los pacientes con ROP preumbral tipo1 de alto riesgo tratados según práctica clínica habitual con bevacizumab intravítreo entre diciembre de 2013 y enero de 2018. Los pacientes con un seguimiento inferior a tres años fueron excluidos. Se registraron los datos de agudeza visual y refracción bajo cicloplejia de la última exploración oftalmológica realizada. Se definió la variable éxito como ausencia de retratamiento con anti-VEGF intravítreo o láser durante el tiempo de seguimiento. Resultados Se incluyeron en el análisis 76 ojos de 38 pacientes. Un total de 20 pacientes (40 ojos) tenían valoración de mejor agudeza visual corregida tomada utilizando la prueba de Snellen. La edad media de estos pacientes era de 6años (intervalo 4-9). La agudeza visual mediana fue de 0,80 (RIQ: 0,50; 1,00). Treinta y cuatro ojos (85%) tenían buena agudeza visual (mayor o igual a 0,5). Se obtuvo la refracción bajo cicloplejia de 74 ojos de 37 pacientes. La mediana del equivalente esférico en la última revisión fue de +0,94 (RIQ: −0,25; 1,88). La tasa de éxito fue del 96,05%. Conclusión El bevacizumab intravítreo es una terapia efectiva con buenos resultados funcionales para ROP tipo1 de alto riesgo. En nuestro estudio se observó buena respuesta al tratamiento, con una tasa de éxito superior al 95% (AU)

Background/aim The aim of the study is to describe the efficacy and to determine the functional outcome in terms of visual acuity and refractive defect of a single dose of intravitreal bevacizumab in patients with high-risk ROP type1. Methods In this retrospective clinical study patients diagnosed between December 2013 and January 2018 with high-risk pre-threshold ROP type1 and treated with intravitreal bevacizumab were selected. All patients were treated following the established protocol at our centre. Those patients with less than three-year follow-up were excluded. Visual acuity and cycloplegic refraction in the last visit were registered. Treatment efficacy was defined as the absence of retreatment with intravitreal anti-VEGF or laser during follow-up. Results A total of 38 infants (76eyes) were included in the analysis. Twenty infants (40eyes) completed visual acuity testing. Mean age was 6years (IQR: 4-9). Median visual acuity was 0.8 (IQR: 0.5-1). Thirty-four eyes (85%) had good visual acuity (greater than or equal to 0.5). Thirty-seven patients (74eyes) had cycloplegic refraction measured. Median spherical equivalent at the last visit was +0.94 (IQR: −0.25; 1.88). Treatment success rate was 96.05%. Conclusion Intravitreal bevacizumab treatment showed good functional outcome in patients with high-risk ROP type1. In our study, good response to treatment was observed with a success rate over 95% (AU)

Functional results in children with retinopathy of prematurity treated with intravitreal bevacizumab.

BACKGROUND/AIM: The aim of the study is to describe the efficacy and to determine the functional outcome in terms of visual acuity and refractive defect of a single dose of intravitreal bevacizumab in patients with high-risk ROP type 1. METHODS: In this retrospective clinical study patients diagnosed between December 2013 and January 2018 with high-risk pre-threshold ROP type 1 and treated with intravitreal bevacizumab were selected. All patients were treated following the established protocol at our centre. Those patients with less than three-year follow-up were excluded. Visual acuity and cycloplegic refraction in the last visit were registered. Treatment efficacy was defined as the absence of retreatment with intravitreal anti-VEGF or laser during follow-up. RESULTS: A total of 38 infants (76 eyes) were included in the analysis. Twenty infants (40 eyes) completed visual acuity testing. Mean age was 6 years (IQR: 4-9). Median visual acuity was 0.8 (IQR: 0.5-1). Thirty-four eyes (85%) had good visual acuity (greater than or equal to 0.5). Thirty-seven patients (74 eyes) had cycloplegic refraction measured. Median spherical equivalent at the last visit was +0.94 (IQR: -0.25; 1.88). Treatment success rate was 96.05%. CONCLUSION: Intravitreal bevacizumab treatment showed good functional outcome in patients with high-risk ROP type 1. In our study, good response to treatment was observed with a success rate over 95%.

Nanoscale Dynamics of Streptococcal Adhesion to AGE-Modified Collagen.

The adhesion of initial colonizers such as Streptococcus mutans to collagen is critical for dentinal and root caries progression. One of the most described pathological and aging-associated changes in collagen-including dentinal collagen-is the generation of advanced glycation end-products (AGEs) such as methylglyoxal (MGO)-derived AGEs. Despite previous reports suggesting that AGEs alter bacterial adhesion to collagen, the biophysics driving oral streptococcal attachment to MGO-modified collagen remains largely understudied. Thus, the aim of this work was to unravel the dynamics of the initial adhesion of S. mutans to type I collagen in the presence and absence of MGO-derived AGEs by employing bacterial cell force spectroscopy with atomic force microscopy (AFM). Type I collagen gels were treated with 10 mM MGO to induce AGE formation, which was characterized with microscopy and enzyme-linked immunosorbent assay. Subsequently, AFM cantilevers were functionalized with living S. mutans UA 159 or Streptococcus sanguinis SK 36 cells and probed against collagen surfaces to obtain force curves displaying bacterial attachment in real time, from which the adhesion force, number of events, Poisson analysis, and contour and rupture lengths for each individual detachment event were computed. Furthermore, in silico computer simulation docking studies between the relevant S. mutans UA 159 collagen-binding protein SpaP and collagen were computed, in the presence and absence of MGO. Overall, results showed that MGO modification increased both the number and adhesion force of single-unbinding events between S. mutans and collagen, without altering the contour or rupture lengths. Both experimental and in silico simulations suggest that this effect is due to increased specific and nonspecific forces and interactions between S. mutans UA 159 and MGO-modified collagen substrates. In summary, these results suggest that collagen alterations due to aging and glycation may play a role in early bacterial adherence to oral tissues, associated with conditions such as aging or chronic hyperglycemia, among others.

Asociación entre morfología del pie y rendimiento en gimnasia rítmica / Association Foot Morphology and Performance in Rhythmic Gymnastics

Los objetivos del estudio fueron determinar la influencia de la morfología del pie en el rendimiento en Gimnasia Rítmica y analizar la influencia de los años de práctica. La muestra estaba compuesta por 48 gimnastas que habían practicado gimnasia federada y competido durante el último año. Los resultados indican que las gimnastas tienen predominantemente un pie neutro y con huella normal, presentando bastante asimetría entre pies, no significativa, lo que puede ser consecuencia de un trabajo unilateral y debería ser corregido en el entrenamiento. Sólo el rango de amplitud de la articulación talocrural parece ser una característica de la morfología del pie que incida sobre el rendimiento técnico y parece más una característica innata. La práctica de la Gimnasia Rítmica podría no ser un factor tan decisivo como podía suponerse en las modificaciones morfológicas de la huella plantar

The objectives of the study were to determine the influence of foot morphology on performance in Rhythmic Gymnastics and to analyze the influence of years of practice. The sample consisted of 48 gymnasts who had practiced federated gymnastics and competed during the last year. The results indicate that the gymnasts have predominantly a neutral foot and with a normal footprint, presenting enough asymmetry between feet, not significant, which may be a consequence of asymmetric work and should be corrected in training. Only the range of amplitude of the talocrural joint seems to be a characteristic of foot morphology that affects technical performance and seems more an innate characteristic. The practice of rhythmic gymnastics might not be as decisive a factor as could be supposed in the morphological modifications of the footprint

Economic valuation of the eco-systemic benefits derived from the environmental asset lake Laguna Santa Elena, through the multi-criteria analysis / Valoração econômica dos benefícios ecossistêmicos derivados do ativo ambiental Laguna Santa Elena por meio da análise multicritério

Abstract Lake Laguna Santa Elena, a freshwater body, located in mid-south of Chile, is an environmental asset used as a water resource by the agricultural and touristic sector and is the habitat for a wide variety of endemic avifauna. The objective of this study was to assign a monetary value to this lacustrine body, using the methodology of AMUVAM (Multicriteria Analysis of Environmental Active Values). The information was collected through the application of surveys to a panel of experts involved in the use of the environmental resource. The result obtained with the AMUVAM method corresponds to the US $ 17,780,686, a figure that represents an estimated value of the TEV (Total Economic Value) of the environmental asset. This study, a pioneer in the Region, will support decision making, allowing an adequate management of this critical water resource.

Resumo A Laguna Santa Elena, um corpo de água doce localizado no centro-sul do Chile, é um ativo ambiental usado como recurso hídrico pelos setores agrícola e turístico e é o habitat de uma ampla variedade de avifauna endêmica. O objetivo deste estudo foi atribuir um valor monetário a esse corpo lacustre, utilizando a metodologia da AMUVAM (Análise Multicritério de Valores Ambientais Ativos). As informações foram coletadas por meio da aplicação de pesquisas a um painel de especialistas envolvidos no uso do recurso ambiental. O resultado obtido com o método AMUVAM corresponde a US$ 17.780.686, que representam um valor estimado do valor econômico total (VT) do ativo ambiental. Este estudo, pioneiro na região, apoiará a tomada de decisões, permitindo um gerenciamento adequado desse recurso hídrico crítico.

Isolation and characterization of Clostridium difficile from a small survey of wastewater, food and animals in New Zealand.

The objective of this study was to undertake a microbiological survey of foods, animal faeces and wastewater samples for Clostridium difficile, and determine the genotypes and antimicrobial susceptibilities of isolates. A total of 211 samples were tested for C. difficile using culture methods. Thirteen toxigenic C. difficile isolates were obtained; ten from wastewater samples, one each from pig and duck faeces and another from a raw meat product. Eight PCR-ribotypes (RTs) were identified, including two novel RTs (878 and 879). Single-nucleotide polymorphism analysis using WGS data for all isolates provided greater discrimination between C. difficile isolates within the same RT and multilocus sequence typing (MLST) profiles. All C. difficile isolates were found to be susceptible to the first-line human antimicrobials used to treat C. difficile infection. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to report the isolation of Clostridium difficile from animals, food and wastewater in New Zealand (NZ) and provides important data with respect to ribotypes and multilocus sequence typing profiles, whole genome sequence and antimicrobial susceptibilities. The results highlight the need for further investigations into the epidemiology of C. difficile in NZ and to elucidate the role of the environmental and food sources as transmission routes of human infection.

Economic valuation of the eco-systemic benefits derived from the environmental asset lake Laguna Santa Elena, through the multi-criteria analysis.

Lake Laguna Santa Elena, a freshwater body, located in mid-south of Chile, is an environmental asset used as a water resource by the agricultural and touristic sector and is the habitat for a wide variety of endemic avifauna. The objective of this study was to assign a monetary value to this lacustrine body, using the methodology of AMUVAM (Multicriteria Analysis of Environmental Active Values). The information was collected through the application of surveys to a panel of experts involved in the use of the environmental resource. The result obtained with the AMUVAM method corresponds to the US $ 17,780,686, a figure that represents an estimated value of the TEV (Total Economic Value) of the environmental asset. This study, a pioneer in the Region, will support decision making, allowing an adequate management of this critical water resource.

An outbreak of multiple genotypes of Listeria monocytogenes in New Zealand linked to contaminated ready-to-eat meats-a retrospective analysis using whole-genome sequencing.

Four cases of listeriosis in a hospital (A) in New Zealand were identified in 2012. Pulsed-field gel electrophoresis (PFGE) used at the time identified four pulsotypes amongst the clinical isolates. Two of the pulsotypes matched to Listeria monocytogenes isolates obtained from ready-to-eat (RTE) meat samples from a RTE producer tested during a nationwide microbiological survey the month prior. The outbreak investigation confirmed that the RTE producer had supplied product to the hospital and additional testing confirmed the presence of L.  monocytogenes in RTE meats from the hospital kitchen. Two further listeriosis cases presented in another hospital (B) with one clinical isolate identified as the same pulsotype as identified for one case in hospital A, but the epidemiology information concluded that the clinical cases from hospital B were not linked to the outbreak. Retrospective whole-genome sequencing confirmed that epidemiologically linked isolates belonging to three different genotypes for clinical cases from hospital A and RTE meats samples from the hospital kitchen differed by 0-1 core-genome locus or single nucleotide polymorphisms (SNP). The use of core-genome multilocus sequence typing and SNP analysis provided a greater degree of discrimination between isolates compared to PFGE. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes a listeriosis outbreak associated with a hospital in New Zealand and attributed to contaminated ready-to-eat (RTE) meat supplied to the hospital by a single producer. Retrospective whole-genome sequence analysis of outbreak isolates was found to provide a greater degree of discrimination between isolates compared to pulsed-field gel electrophoresis and supported the conclusions made at the time of the outbreak. The multiple genotypes identified from clinical cases and the RTE meats obtained during the outbreak highlight the importance of epidemiological concordance alongside genotyping.

Changes in Corneal Expression of MUC5AC after Autologous Serum Eyedrop Treatment in Patients with Limbal Stem Cell Deficiency.

Purpose: To evaluate the effect of autologous serum eyedrops treatment on corneal expression of the MUC5AC in patients with limbal deficiency. Methods: A prospective and comparative interventional case series study of 42 eyes of 21 patients was performed before and 8 weeks after treatment with autologous serum. All patients underwent a complete ophthalmic assessment of the tear film and ocular surface, corneal impression cytology (IC) and MUC5AC detection by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Results: Forty-one of the 42 eyes studied were available for both conventional cytology and MUC5AC analysis prior to and after treatment. Differences between outcomes obtained by impression cytology and MUC5AC detection were found in 9 of 82 samples (11%). We found changes in the corneal expression of MUC5AC after treatment in 19 of 41 eyes (46.3%): 18 of them (94.7%) changed from positive to negative expression, and 1 eye (5.3%) changed from negative to positive MUC5AC expression after autologous serum eyedrops. These changes were related with the corneal involvement prior to treatment (15 of them (78.9%) occurred in patients with slight corneal involvement), and with the improvement in the degree of squamous metaplasia after treatment (P = .001 and P = .003, respectively). Conclusions: The treatment significantly improved tear stability, squamous metaplasia, and subjective patient perception. Autologous serum eyedrops treatment diminished the corneal expression of MUC5AC mainly in patients with slight corneal involvement before treatment.

Parotid Salivary Gland Basal Cell Adenocarcinoma in a Big-eared Opossum (Didelphis aurita).

The opossum (family Didelphidae) is a marsupial endemic to the Americas. Apart from the South American short-tailed opossum (Monodelphis domestica) and the Virginia opossum (Didelphis virginiana), there is considerable lack of knowledge about the health and diseases of most opossum species. Among these, the big-eared opossum (Didelphis aurita) is found in Argentina, Brazil and Paraguay. Natural and experimental studies have shown this species to be susceptible to infectious agents with zoonotic potential and the animals may play a role in transmission of such agents. However, neoplasia appears to be uncommon in this species. We describe the gross, microscopical and immunohistochemical features of a parotid salivary gland basal cell adenocarcinoma in a free-living big-eared opossum. This case represents the first report of salivary gland neoplasia in opossums.

Microbial competition in environmental nosocomial reservoirs and diffusion capacity of OXA48-Klebsiella pneumoniae: potential impact on patients and possible control methods.

BACKGROUND: We have found clusters of Klebsiella pneumoniae with OXA48-carbepenemase cases in some hospital rooms, and decided to investigate whether bathroom siphons could be a reservoir for OXA48 bacteria, as occurs with K. oxytoca with other types of carbepenemases. METHODS: We evaluated the microbial competition between strains with OXA48 and VIM carbepenemases, in diluted nutrient-broth, on a slime germ-carrier. We compared the number of colonies at 5 and 10 days on the contaminated carriers with one or two strains. We evaluated the dissemination of K. pneumoniae with carbepenemase OXA48 or VIM from thumbs and index fingers of volunteers, to standard surfaces (20 glass germ-carrier by each volunteer). After, we counted the number of microorganisms on each carrier. Microbiological weekly studies of faecal microbiota of all patients were obtained in Traumatology and Oncology. Moreover, we studied samples of the sink in their rooms. PCR and MLST sequence-type was determined in all K. pneumoniae diagnosed from patients and sinks. RESULTS: A large possibility of diffusion from contaminated hands, which continue to transmit high numbers of microorganisms after more than 10 successive surface contacts, was highlighted; OXA bacteria were more persistent than VIM bacteria. Microbial competition studies showed that VIM bacteria are inhibited by OXA ones. These observations can explain the concentration of cases of K. pneumoniae OXA48 in some rooms in Traumatology and Oncology, producing a significant OR between rooms with OXA48-bacteria-contaminated siphons and other rooms (3.1 and 3.3 respectively). Risk was lowered after changing or disinfecting (heat plus chlorinated disinfectant) the contaminated siphons. Siphon colonization by VIM bacteria was not related with human infections by similar microorganisms. CONCLUSIONS: Bathroom siphons can be a reservoir for K. pneumoniae OXA48 and lead to outbreaks. Outbreaks can be controlled by replacement or heat plus chemical treatment of the sink-siphons.

Genetic characterization of a mouse line with primary aldosteronism.

In an attempt to define novel genetic loci involved in the pathophysiology of primary aldosteronism, a mutagenesis screen after treatment with the alkylating agent N-ethyl-N-nitrosourea was established for the parameter aldosterone. One of the generated mouse lines with hyperaldosteronism was phenotypically and genetically characterized. This mouse line had high aldosterone levels but normal creatinine and urea values. The steroidogenic enzyme expression levels in the adrenal gland did not differ significantly among phenotypically affected and unaffected mice. Upon exome sequencing, point mutations were identified in seven candidate genes (Sspo, Dguok, Hoxaas2, Clstn3, Atm, Tipin and Mapk6). Subsequently, animals were stratified into wild-type and mutated groups according to their genotype for each of these candidate genes. A correlation of their genotypes with the respective aldosterone, aldosterone-to-renin ratio (ARR), urea and creatinine values as well as steroidogenic enzyme expression levels was performed. Aldosterone values were significantly higher in animals carrying mutations in four different genes (Sspo, Dguok, Hoxaas2 and Clstn3) and associated statistically significant adrenal Cyp11b2 overexpression as well as increased ARR was present only in mice with Sspo mutation. In contrast, mutations of the remaining candidate genes (Atm, Tipin and Mapk6) were associated with lower aldosterone values and lower Hsd3b6 expression levels. In summary, these data demonstrate association between the genes Sspo, Dguok, Hoxaas2 and Clstn3 and hyperaldosteronism. Final proofs for the causative nature of the mutations have to come from knock-out and knock-in experiments.

Economic valuation of the eco-systemic benefits derived from the environmental asset lake Laguna Santa Elena, through the multi-criteria analysis

Abstract Lake Laguna Santa Elena, a freshwater body, located in mid-south of Chile, is an environmental asset used as a water resource by the agricultural and touristic sector and is the habitat for a wide variety of endemic avifauna. The objective of this study was to assign a monetary value to this lacustrine body, using the methodology of AMUVAM (Multicriteria Analysis of Environmental Active Values). The information was collected through the application of surveys to a panel of experts involved in the use of the environmental resource. The result obtained with the AMUVAM method corresponds to the US $ 17,780,686, a figure that represents an estimated value of the TEV (Total Economic Value) of the environmental asset. This study, a pioneer in the Region, will support decision making, allowing an adequate management of this critical water resource.

Resumo A Laguna Santa Elena, um corpo de água doce localizado no centro-sul do Chile, é um ativo ambiental usado como recurso hídrico pelos setores agrícola e turístico e é o habitat de uma ampla variedade de avifauna endêmica. O objetivo deste estudo foi atribuir um valor monetário a esse corpo lacustre, utilizando a metodologia da AMUVAM (Análise Multicritério de Valores Ambientais Ativos). As informações foram coletadas por meio da aplicação de pesquisas a um painel de especialistas envolvidos no uso do recurso ambiental. O resultado obtido com o método AMUVAM corresponde a US$ 17.780.686, que representam um valor estimado do valor econômico total (VT) do ativo ambiental. Este estudo, pioneiro na região, apoiará a tomada de decisões, permitindo um gerenciamento adequado desse recurso hídrico crítico.

Genomic Appraisal of the Multifactorial Basis for In Vitro Acquisition of Miltefosine Resistance in Leishmania donovani.

Protozoan parasites of the Leishmania donovani complex are the causative agents of visceral leishmaniasis (VL), the most severe form of leishmaniasis, with high rates of mortality if left untreated. Leishmania parasites are transmitted to humans through the bite of infected female sandflies (Diptera: Phlebotominae), and approximately 500,000 new cases of VL are reported each year. In the absence of a safe human vaccine, chemotherapy, along with vector control, is the sole tool with which to fight the disease. Miltefosine (hexadecylphosphatidylcholine [HePC]), an antitumoral drug, is the only successful oral treatment for VL. In the current study, we describe the phenotypic traits of L. donovani clonal lines that have acquired resistance to HePC. We performed whole-genome and RNA sequencing of these resistant lines to provide an inclusive overview of the multifactorial acquisition of experimental HePC resistance, circumventing the challenge of identifying changes in membrane-bound proteins faced by proteomics. This analysis was complemented by assessment of the in vitro infectivity of HePC-resistant parasites. Our work underscores the importance of complementary "omics" to acquire the most comprehensive insight for multifaceted processes, such as HePC resistance.

Genetics of Cushing's disease: an update.

INTRODUCTION: Cushing's disease (CD) results from uncontrolled hypercortisolism induced by ACTH-secreting corticotroph adenomas; accordingly, patients diagnosed with CD usually present several comorbidities and an increased risk of mortality. Hypothesis-driven screenings have led to identification of rare alterations in a low number of patients, although the genetic basis underlying CD has remained unclear until recently. Using whole-exome sequencing, recurrent mutations have been reported in the gene coding for the ubiquitin-specific protease 8 (USP8), a protein with deubiquitinase (DUB) activity that modulates the lysosomal turnover of the EGF receptor (EGFR) and other membrane proteins. METHODS: In this review, we summarize the recent genetic findings and discuss the clinical and pathological implications of USP8 deregulation in corticotroph adenomas. CONCLUSIONS: Mutations in USP8 have been identified in 35-62 % of functional sporadic corticotroph adenomas causing Cushing's disease, but not in any other type of pituitary tumor. These mutations are found mostly in adult female patients and lead to an aberrant DUB activation by impairing the regulation of USP8 by members of the 14-3-3 family of proteins. The consequence of this hyperactivation is a longer retention of EGFR at the plasma membrane which promotes an enhanced production of ACTH.

A DNA Aptasensor for Electrochemical Detection of Vascular Endothelial Growth Factor.

Electrochemical aptasensors can detect different cancer biomarkers to provide point-of-care diagnosis that is low cost, rapid, specific and sensitive. In this work, we described the development of an electrochemical single-use aptasensor for detection and analysis of vascular endothelial growth factor (VEGF). Gold nanostructured graphite screen-printed electrodes were firstly modified with a mixed monolayer of a primary thiolated DNA aptamer and a spacer thiol, 6-mercapto-1-hexanol. VEGF protein was then incubated with the aptasensor. An enzyme-amplified detection scheme, based on the coupling of a streptavidin-alkaline phosphatase conjugate and secondary biotinylated aptamer was then applied. The enzyme catalyzed the hydrolysis of the electroinactive 1 -naphthyl-phosphate to 1 -naphthol; this product is electroactive and has been detected by means of differential pulse voltammetry (DPV). The aptasensor response was found to be linearly related to the target concentration between 0 and 250 nmol L(-1); the detection limit was 30 nmol L(-1). The performance of the immunoassay in terms of reproducibility and selectivity has been also studied.

Encefalopatía tardía por monóxido de carbono de curso reversible en un paciente tratado con citicolina / Delayed-onset encephalopathy caused by carbon monoxide poisoning: Resolution in a patient treated with citicoline

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Caracterización del cultivo primario epitelial de conjuntiva humana / Characterization of epithelial primary culture from human conjunctiva

OBJETIVO: Evaluar diferentes cultivos primarios de conjuntiva humana enriquecidos con suero fetal bovino, suero autólogo y suero autólogo rico en plaquetas, sobre un soporte de membrana amniótica humana y cápsula anterior del cristalino. MÉTODOS: Ciento cuarenta y ocho explantes de conjuntiva humana fueron cultivados en CnT50® enriquecido con 1, 2,5, 5 y 10% en suero fetal bovino, suero autólogo y suero autólogo rico en plaquetas. Las muestras de conjuntivales fueron incubadas a 37 °C, 5% CO2 y 95% HR, durante 3 semanas. RESULTADOS: Todos los cultivos primarios tuvieron el fenotipo típico de las células epiteliales de la conjuntiva. Los cultivos mostraron células secretoras MUC5AC-positiva, células conjuntivales CK19-positiva y células conjuntivales no secretoras MUC4-positiva, pero sin fenotipo corneal (CK3-negativa) ni fibroblastos (CD90-negativa). CONCLUSIONES: Las células epiteliales progenitoras de la conjuntiva se mantuvieron en todos los cultivos, por lo que un cultivo celular en CnT50® enriquecido con suero autólogo entre el 1 y 5% sobre membrana amniótica humana puede proporcionar la mejor información de la diferenciación de las células epiteliales para la reconstrucción de la superficie conjuntival

OBJECTIVE: To evaluate primary cultures from human conjunctiva supplemented with fetal bovine serum, autologous serum, and platelet-rich autologous serum, over human amniotic membrane and lens anterior capsules. METHODS: One-hundred and forty-eight human conjunctiva explants were cultured in CnT50® supplemented with 1, 2.5, 5 and 10% fetal bovine serum, autologous serum and platelet-rich autologous serum. Conjunctival samples were incubated at 37 °C, 5% CO2 and 95% HR, for 3 weeks. RESULTS: The typical phenotype corresponding to conjunctival epithelial cells was present in all primary cultures. Conjunctival cultures had MUC5AC-positive secretory cells, K19-positive conjunctival cells, and MUC4-positive non-secretory conjunctival cells, but were not corneal phenotype (cytokeratin K3-negative) and fibroblasts (CD90-negative). CONCLUSIONS: Conjunctiva epithelial progenitor cells were preserved in all cultures; thus, a cell culture in CnT50® supplemented with 1 to 5% autologous serum over human amniotic membrane can provide better information of epithelial cell differentiation for the conjunctival surface reconstruction