24 - Ciències de la Vida

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    Standardizing in vitro β-lactam antibiotic allergy testing with synthetic IgE
    (American Chemical Society (ACS), 2023) Quintero Campos, Pedro; Gozalbo Rovira, Roberto Vicente; Rodríguez Díaz, Jesús; Maquieira, Ángel; Morais Ezquerro, Sergi Beñat
    The global prevalence of β-lactam allergy poses a major challenge in administering first-line antibiotics, such as penicillins, to a significant portion of the population. The lack of β-lactam IgE antibody pools with defined selectivity hampers the standardization and validation of in vitro assays for β-lactam allergy testing. To address this limitation, this study introduces a synthetic IgE specific to β-lactam antibiotics as a valuable tool for drug allergy research and diagnostic tests. Using phage display technology, we constructed a library of human single-chain antibody fragments (scFv) to target the primary determinant of amoxicillin, a widely used β-lactam antibiotic. Subsequently, we produced a complete human synthetic IgE molecule using the highly efficient baculovirus expression vector system. This synthetic IgE molecule served as a standard in an in vitro chemiluminescence immunoassay for β-lactam antibiotic allergy testing. Our results demonstrated a detection limit of 0.05 IU/mL (0.63 pM), excellent specificity (100%), and a four-fold higher clinical sensitivity (73%) compared to the in vitro reference assay when testing a cohort of 150 serum samples. These findings have significant implications for reliable interlaboratory comparison studies, accurate labeling of allergic patients, and combating the global public health threat of antimicrobial resistance. Furthermore, by serving as a valuable trueness control material, the synthetic IgE facilitates the standardization of diagnostic tests for β-lactam allergy and demonstrates the potential of utilizing this synthetic strategy as a promising approach for generating reference materials in drug allergy research and diagnostics.
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    Culture of human rotaviruses in relevant models shows differences in culture-adapted and nonculture-adapted strains
    (MDPI AG, 2023) Peña Gil, Nazaret; Randazzo, Walter; Carmona Vicente, Noelia; Santiso Bellón, Cristina; Cárcamo-Calvo, Roberto; Navarro Lleó, Noemí; Monedero, Vicente; Yebra Yebra, María Jesús; Buesa, Julia; Gozalbo Rovira, Roberto Vicente; Rodríguez Díaz, Jesús
    Rotavirus (RV) is the leading cause of acute gastroenteritis (AGE) in children under 5 years old worldwide, and several studies have demonstrated that histo-blood group antigens (HBGAs) play a role in its infection process. In the present study, human stool filtrates from patients diagnosed with RV diarrhea (genotyped as P[8]) were used to infect differentiated Caco-2 cells (dCaco-2) to determine whether such viral strains of clinical origin had the ability to replicate in cell cultures displaying HBGAs. The cell culture-adapted human RV Wa model strain (P[8] genotype) was used as a control. A time-course analysis of infection was conducted in dCaco-2 at 1, 24, 48, 72, and 96 h. The replication of two selected clinical isolates and Wa was further assayed in MA104, undifferentiated Caco-2 (uCaco-2), HT29, and HT29-M6 cells, as well as in monolayers of differentiated human intestinal enteroids (HIEs). The results showed that the culture-adapted Wa strain replicated more efficiently in MA104 cells than other utilized cell types. In contrast, clinical virus isolates replicated more efficiently in dCaco-2 cells and HIEs. Furthermore, through surface plasmon resonance analysis of the interaction between the RV spike protein (VP8*) and its glycan receptor (the H antigen), the V7 RV clinical isolate showed 45 times better affinity compared to VP8* from the Wa strain. These findings support the hypothesis that the differences in virus tropism between clinical virus isolates and RV Wa could be a consequence of the different HBGA contents on the surface of the cell lines employed. dCaco-2, HT29, and HT29M6 cells and HIEs display HBGAs on their surfaces, whereas MA104 and uCaco-2 cells do not. These results indicate the relevance of using non-cell culture-adapted human RV to investigate the replication of rotavirus in relevant infection models.
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    SARS-CoV-2 N-antigenemia in critically ill adult COVID-19 patients: frequency and association with inflammatory and tissue-damage biomarkers
    (Wiley, 2022) Olea Miguel, Beatriz; Albert Vicent, E.; Rodríguez Díaz, Jesús; Gozalbo Rovira, Roberto Vicente; Navarro, David; Blasco, María Luisa
    The current study aimed at characterizing the dynamics of SARS-CoV-2 nucleocapsid (N) antigenemia in a cohort of critically ill adult COVID-19 patients and assessing its potential association with plasma levels of biomarkers of clinical severity and mortality. Seventy-three consecutive critically ill COVID-19 patients (median age, 65 years) were recruited. Serial plasma (n = 340) specimens were collected. A lateral flow immunochromatography assay and reverse-transcription polymerase chain reaction (RT-PCR) were used for SARS-CoV-2 N protein detection and RNA quantitation and in plasma, respectively. Serum levels of inflammatory and tissue-damage biomarkers in paired specimens were measured. SARS-CoV-RNA N-antigenemia and viral RNAemia were documented in 40.1% and 35.6% of patients, respectively at a median of 9 days since symptoms onset. The level of agreement between the qualitative results returned by the N-antigenemia assay and plasma RT-PCR was moderate (k = 0.57; p < 0.0001). A trend towards higher SARS-CoV-2 RNA loads was seen in plasma specimens testing positive for N-antigenemia assay than in those yielding negative results (p = 0.083). SARS-CoV-2 RNA load in tracheal aspirates was significantly higher (p < 0.001) in the presence of concomitant N-antigenemia than in its absence. Significantly higher serum levels of ferritin, lactose dehydrogenase, C-reactive protein, and D-dimer were quantified in paired plasma SARS-CoV-2 N-positive specimens than in those testing negative. Occurrence of SARS-CoV-2 N-antigenemia was not associated with increased mortality in univariate logistic regression analysis (odds ratio, 1.29; 95% confidence interval, 0.49-3.34; p = 0.59). In conclusion, SARS-CoV-2 N-antigenemia detection is relatively common in ICU patients and appears to associate with increased serum levels of inflammation and tissue-damage markers. Whether this virological parameter may behave as a biomarker of poor clinical outcome awaits further investigations.
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    The life of saccharomyces and con-saccharomyces Yas in drinking wine
    (MDPI AG, 2023) Maicas, Sergi; Mateo, José Juan
    Drinking wine is a processed beverage that offers high nutritional and health benefits. It is produced from grape must, which undergoes fermentation by yeasts (and sometimes lactic acid bacteria) to create a product that is highly appreciated by consumers worldwide. However, if only one type of yeast, specifically Saccharomyces cerevisiae, was used in the fermentation process, the resulting wine would lack aroma and flavor and may be rejected by consumers. To produce wine with a desirable taste and aroma, non-Saccharomyces yeasts are necessary. These yeasts contribute volatile aromatic compounds that significantly impact the wine's final taste. They promote the release of primary aromatic compounds through a sequential hydrolysis mechanism involving several glycosidases unique to these yeasts. This review will discuss the unique characteristics of these yeasts (Schizosaccharomyces pombe, Pichia kluyveri, Torulaspora delbrueckii, Wickerhamomyces anomalus, Metschnikowia pulcherrima, Hanseniaspora vineae, Lachancea thermotolerans, Candida stellata, and others) and their impact on wine fermentations and co-fermentations. Their existence and the metabolites they produce enhance the complexity of wine flavor, resulting in a more enjoyable drinking experience.
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    Assessment of pollution along the Northern Iberian shelf by the combined use of chemical and biochemical markers in two representative fish species
    (Elsevier, 2008) Fernandes, Denise; Andreu-Sánchez, Oscar; Bebianno, Maria Joao; Porte, Cinta
    Muscle concentrations of organochlorinated compounds as well as biliary levels of polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APEs) were determined in two different fish species, the four-spotted megrim (Lepidorhombus boscii) and the pouting (Trisopterus luscus) collected along the Northern Iberian coast. Additionally, a set of biochemical markers namely, 7-ethoxyresorufin O-deethylase (EROD), UDP-glucuronosyltransferase (UGT) and catalase (CAT) were measured in liver subcellular fractions. Chemical analysis indicated geographical differences in pollutant loads that were further reinforced by biomarker responses. Thus, EROD activity showed a good correlation with the amount of PCBs bioaccumulated in muscle tissue of both fish species. Elevated UGT activity was observed in those individuals highly exposed to APEs and 1-naphthol. The study reinforces the need to select representative sentinel species from different habitats for biomonitoring purposes and provides further support for the use of biomarkers in assessing the health of coastal areas.
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    Respuesta inmune celular en la hepatitis C
    (Elsevier, 2001) López Labrador, F. Xavier
    Respuesta inmune celular en la hepatitis C
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    Severe recurrent hepatitis C following re-transplantation for HCV-related graft cirrhosis
    (2003) Berenguer, Marina; Prieto, M.; Palau, A.; Rayón, J. M.; Carrasco, D.; San Juan, F.; López Labrador, F. Xavier; Moreno, R.; Mir, J.; Berenguer, J.
    An increase in the number of hepatitis C virus (HCV)-infected transplant recipients at need for repeated liver transplantation is anticipated. To date, there is a certain reluctance to accept these patients because of an increased organ shortage, early reports suggesting a poor outcome, and uncertainty regarding the natural history of recurrent hepatitis C in the second graft. The aim of this study is to determine the outcome of patients undergoing retransplantation for HCV-related graft cirrhosis. Of 49 transplant recipients with HCV-related allograft cirrhosis, 31 patients developed decompensation with criteria for retransplantation. Thirteen patients were denied this option. Of the 18 patients accepted, 6 patients died while on the waiting list (5 patients died of graft cirrhosis at a median of 3.2 months of listing), and 12 patients have undergone retransplantation (median, 10 months since HCV cirrhosis). After retransplantation, 8 patients (67%) died at a median of 8 months, and 4 patients (33%) remain alive after 1.9 years of follow-up. Causes and times of death from retransplantation were: surgical complications, n = 3 (perioperative period); HCV cirrhosis of the second graft, n = 2 (at 9 and 54 months); fibrosing cholestatic hepatitis, n = 1 (at 2 years); lymphoproliferative disorder, n = 1 (at 7 months); and endocarditis, n = 1 (at 3.5 years, with underlying cirrhosis). Of the 4 patients alive, fibrosis stages in the last biopsy specimens are stage 1 (n = 1), stage 3 (n = 1), and stage 4 or cirrhosis (n = 1; one patient has not undergone biopsy), despite antiviral therapy. The outcome of retransplantation for HCV cirrhosis of the first graft is very poor because of multiple complications. The severity of recurrent HCV disease in the second graft seems to be related to that observed in the first graft.
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    Clinical relevance of genetic heterogeneity in HCV
    (Future Science Group, 2010) González-Candelas, F.; López Labrador, F. Xavier
    Clinical relevance of genetic heterogeneity in HCV
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    Influenza epidemiology and influenza vaccine effectiveness during the 2014-2015 season: annual report from the Global Influenza Hospital Surveillance Network
    (2016) Puig-Barberà, J.; Burtseva, E.; Yu, H.; Cowling, B. J.; Badur, S.; Kyncl, J.; Sominina, A.; Afanasieva, O.; Afanasieva, V.; Ciblak, M.; Aktas, F.; Badur, S.; Belenguer-Varea, Á.; Borekci, S.; Boza, F.; Bursteva, E.; Buzitskaya, Z.; Caetano, B.; Cai, J.; Çakir, B.; Carballido-Fernández, M.; Carbonell-Franco, E.; Carratalá-Munuera, C.; Çelebi, S.; Chai, C.; de Paula Ide, L.; Chen, E.; Cowling, B.; Cui, Y.; Deniz, D.; Dondurei, E.; Dong, H.; Dong, X.; Durusu, M.; El Guerche-Séblain, C.; Enda-Moura, F.; Eren, Ş.; Ensoy, A.; Fadeev, A.; Feng, L.
    Background: The Global Influenza Hospital Surveillance Network (GIHSN) aims to determine the burden of severe influenza disease and Influenza Vaccine Effectiveness (IVE). This is a prospective, active surveillance and hospital-based epidemiological study to collect epidemiological data in the GIHSN. In the 2016-2017 influenza season, 15 sites in 14 countries participated in the GIHSN, although the analyses could not be performed in 2 sites. A common core protocol was used in order to make results comparable. Here we present the results of the GIHSN 2016-2017 influenza season. Methods: A RT-PCR test was performed to all patients that accomplished the requirements detailed on a common core protocol. Patients admitted were included in the study after signing the informed consent, if they were residents, not institutionalised, not discharged in the previous 30 days from other hospitalisation with symptoms onset within the 7 days prior to admission. Patients 5 years old or more must also complied the Influenza-Like Illness definition. A test negative-design was implemented to perform IVE analysis. IVE was estimated using a logistic regression model, with the formula IVE = (1-aOR) × 100, where aOR is the adjusted Odds Ratio comparing cases and controls. Results: Among 21,967 screened patients, 10,140 (46.16%) were included, as they accomplished the inclusion criteria, and tested, and therefore 11,827 (53.84%) patients were excluded. Around 60% of all patients included with laboratory results were recruited at 3 sites. The predominant strain was A(H3N2), detected in 63.6% of the cases (1840 patients), followed by B/Victoria, in 21.3% of the cases (618 patients). There were 2895 influenza positive patients (28.6% of the included patients). A(H1N1)pdm09 strain was mainly found in Mexico. IVE could only be performed in 6 sites separately. Overall IVE was 27.24 (95% CI 15.62-37.27. Vaccination seemed to confer better protection against influenza B and in people 2-4 years, or 85 years old or older. The aOR for hospitalized and testing positive for influenza was 3.02 (95% CI 1.59-5.76) comparing pregnant with non-pregnant women. Conclusions: Vaccination prevented around 1 in 4 hospitalisations with influenza. Sparse numbers didn't allow estimating IVE in all sites separately. Pregnancy was found a risk factor for influenza, having 3 times more risk of being admitted with influenza for pregnant women.
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    Relating the liver damage with hepatitis C virus polymorphism in core región and human variables in HIV-1 coinfected patients.
    (2010) Matas, M.; Picornell, A.; Cifuentes, C.; Payeras, A.; Bassa, A.; Homar, F.; López Labrador, F. Xavier; Moya, A., Ramon, M. M.; Castro, J. A.
    La coinfección por el virus de la inmunodeficiencia humana (VIH) y los virus de la hepatitis B (VHB) y C (VHC) es un hecho frecuente, debido fundamentalmente a que comparten las mismas vías de transmisión. Se estima que la coinfección por estos virus se sitúa entre un 5 y un 95%, según el virus de la hepatitis y la población de riesgo considerados, de tal forma que la infección por el VHC es relativamente infrecuente entre los homosexuales infectados por el VIH, mientras que es prácticamente universal en los hemofílicos VIH positivos.
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    Characterization of a Novel Conformational GII.4 Norovirus Epitope: Implications for Norovirus-Host Interactions.
    (2016) Carmona Vicente, Noelia; Vila Vicent, Susana; Allen, D.; Gozalbo-Rovira, R.; Iturriza-Gómara, M.; Buesa, J.; Rodríguez-Díaz, J.
    Characterization of a Novel Conformational GII.4 Norovirus Epitope: Implications for Norovirus-Host Interactions.
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    Genomic medicine reaches HCV-related liver transplantation: hopes and clinical and Public Health implications.
    (2011) López Labrador, F. Xavier; Berenguer, Marina
    Genomic medicine reaches HCV-related liver transplantation: hopes and clinical and Public Health implications
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    Hepatitis C virus resistance to new specifically-targeted antivial therapy (STAT-C): a Public Health perspective
    (Baishideng Publishing Group, 2013) Salvatierra, Karina; Fareleski, Sabrina; Forcada, Alicia; López Labrador, F. Xavier
    Until very recently, treatment for chronic hepatitis C virus (HCV) infection has been based on the combination of two non-viral specific drugs: pegylated interferon-α and ribavirin, which is effective in, overall, about 40%-50% of cases. To improve the response to treatment, novel drugs have been designed to specifically block viral proteins. Multiple compounds are under development, and the approval for clinical use of the first of such direct-acting antivirals in 2011 (Telaprevir and Boceprevir), represents a milestone in HCV treatment. HCV therapeutics is entering a new expanding era, and a highly-effective cure is envisioned for the first time since the discovery of the virus in 1989. However, any antiviral treatment may be limited by the capacity of the virus to overcome the selective pressure of new drugs, generating antiviral resistance. Here, we try to provide a basic overview of new treatments, HCV resistance to new antivirals and some considerations derived from a Public Health perspective, using HCV resistance to protease and polymerase inhibitors as examples.
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    Hepatitis C virus viral kinetics during α-2a or α-2b pegylated interferon plus ribavirin therapy in liver transplant recipients with different immunosuppression regimes
    (2012) Berenguer, Marina; Ortiz-Cantó, C.; Abellán, J. J.; Aguilera, V.; Rubín, A.; Prieto, M .; López Labrador, F. Xavier
    Background: Predictors of sustained virological response (SVR) to antiviral therapy post-liver transplantation (LT) for chronic hepatitis C are needed. In non-transplanted patients, viral kinetics can predict SVR. Objectives: To determine the early viral kinetics in LT recipients with different immunosuppression (tacrolimus - Tac- vs. cyclosporine - CsA-) during treatment with peg-IFN+RBV. Study design: Prospective pilot study in HCV-1b infected patients: (LT CsA n=8; Tac n=8; non-LT n=4), treated with IFN α-2a vs. α-2b (180 μg or 1.5 μg/kg, respectively) once weekly plus weight-based RBV. Median CsA or Tac baseline trough levels were 141 and 7.70 ng/mL, respectively. HCV-RNA was quantified before treatment and after 3, 6, 12h; days 1-6; and weeks 4, 12, 24, 48 and 78 (follow-up). Results: Different kinetics were observed: early viral load declines with shoulder phase (n=12), delayed monophasic without first phase (n=5, all CsA), and biphasic (n=1) or flat (n=1), without influence of IL28B rs12979860 donor/recipient alleles. In LT, median declines (log(10)UI/mL) at week 4 were -3.62 and -1.49 for Tac vs. CsA; and -2.10 vs.-1.50 for IFN α-2a vs. α-2b (NS), with a trend for faster declines in Tac patients. Generalized additive models suggested a cut-off for predicting response in LT patients of 30 days for Tac, but beyond day 40 for CsA. Conclusion: In LT, the viral kinetics during peg-IFN+RBV treatment is delayed. HCV-RNA at 48 h. may not be predictive of response, and CsA-immunosupressed patients with delayed monophasic declines may potentially achieve ETVR and SVR despite unfavourable or absent early viral load declines.
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    Memoria en las cuasiespecies víricas
    (2001) López Labrador, F. Xavier
    Memoria en las cuasiespecies víricas
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    Generalized Linear Model (GLM) framework for the association of host variables and viral strains with liver fibrosis in HCV/HIV coinfected patients.
    (2013) Matas, M.; Picornell, A.; Cifuentes, C.; Payeras, A.; Bassa, A.; Homar, F.; González-Candelas, F.; López Labrador, F. Xavier; Moya, A.; Ramon, M.M.; Castro, J.A.
    Chronic hepatitis C virus (HCV) infection is the main cause of advanced and end-stage liver disease world-wide, and an important factor of morbidity and mortality in Human Immunodeficiency virus-1 (HIV-1) co-infected individuals. Whereas the genetic variability of HCV has been studied extensively in monoinfected patients, comprehensive analyses of both patient and virus characteristics are still scarce in HCV/HIV co-infection. In order to find correlates for liver damage, we sought to analyze demographic, epidemiological and clinical features of HCV/HIV co-infected patients along with the genetic makeup of HCV (viral subtypes and lineage studied by nucleotide sequencing and phylogenetic analysis of the NS5B region). We used the Generalized Linear Model (GLM) methodology in order to integrate data from the virus and the infected host to find predictors for liver damage. The degree of liver disease was evaluated indirectly by means of two indexes (APRI and FIB-4) and accounting for the time since infection, to estimate fibrosis progression rates. Our analyses identified a reduced number of variables (both from the virus and the host) implicated in liver damage, which included the stage of HIV infection, levels of gamma-glutamil transferase and cholesterol, and some distinct HCV phylogenetic clades.
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    Is PD-1 blockade a potential therapy for HBV?
    (2019) Féray, C.; López Labrador, F. Xavier
    HBV tolerance is partially understood. HBV is a non-cytopathic virus and its pathogenesis lies in immune-mediated liver injury1 . Most people in the HBsAg-positive population are infected as children. For years or decades, HBV is tolerated with high levels of viral replication. The role of HBeAg2 , the age of infection and the liver environment3 all appear to be crucial features of this specific immunotolerance. After this stage, immunotolerance and viral replication decrease, HBeAg escape mutations occur and a frequent scenario is HBsAg persistence, low replication and mild or no liver disease. Subsequently HBsAg can be cleared, or liver disease may progress to chronic hepatitis and cirrhosis. The main complication of HBV infection is hepatocellular carcinoma (HCC). Although oncogenic pathways are related to the life cycle of HBV (transactivation by the HBx protein and DNA viral integration into the liver genome), one very important risk factor for the development of HCC is liver inflammation and regeneration, both of which are mediated by adaptive immunity
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    Boceprevir in the treatment of chronic hepatitis C virus infection.
    (2011) M Berenguer; FX López-Labrador
    Boceprevir in the treatment of chronic hepatitis C virus infection.
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    Recommendations for enterovirus diagnostics and characterisation within and beyond Europe.
    (2018) Harvala, H.; Broberg, E.; Benschop, K.; Berginc N; Ladhani S; Susi P; Christiansen C; McKenna J; Allen D; Makiello P; McAllister G; Carmen M; Zakikhany K; Dyrdak R; Nielsen X; Madsen T; Paul J; Moore C; von Eije K; Piralla A; Carlier M; Vanoverschelde L; Poelman R; Anton A; López Labrador, F. Xavier ; Pellegrinelli L; Keeren K; Maier M; Cassidy H; Derdas S; Savolainen-Kopra C; Diedrich S; Nordbø S; Buesa J; Bailly JL; Baldanti F; MacAdam A; Mirand A; Dudman S; Schuffenecker I
    Enteroviruses (EV) can cause severe neurological and respiratory infections, and occasionally lead to devastating outbreaks as previously demonstrated with EV-A71 and EV-D68 in Europe. However, these infections are still often underdiagnosed and EV typing data is not currently collected at European level. In order to improve EV diagnostics, collate data on severe EV infections and monitor the circulation of EV types, we have established European non-polio enterovirus network (ENPEN). First task of this cross-border network has been to ensure prompt and adequate diagnosis of these infections in Europe, and hence we present recommendations for non-polio EV detection and typing based on the consensus view of this multidisciplinary team including experts from over 20 European countries. We recommend that respiratory and stool samples in addition to cerebrospinal fluid (CSF) and blood samples are submitted for EV testing from patients with suspected neurological infections. This is vital since viruses like EV-D68 are rarely detectable in CSF or stool samples. Furthermore, reverse transcriptase PCR (RT-PCR) targeting the 5'noncoding regions (5'NCR) should be used for diagnosis of EVs due to their sensitivity, specificity and short turnaround time. Sequencing of the VP1 capsid protein gene is recommended for EV typing; EV typing cannot be based on the 5'NCR sequences due to frequent recombination events and should not rely on virus isolation. Effective and standardized laboratory diagnostics and characterisation of circulating virus strains are the first step towards effective and continuous surveillance activities, which in turn will be used to provide better estimation on EV disease burden.
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    Tools for Nutrition Assessment of Adults with Cerebral Palsy: Development of a Gold Standard
    (2023) Expósito, D.; Morales-Suarez, M.M.; Soriano, J.M.; Soler, C.
    Purpose of Review Cerebral palsy (CP) is a group of disorders caused by non-stabilized cerebral lesions. Individuals with this disorder are at a higher risk of suffering from malnutrition and other related detrimental effects to their quality of life. For this reason, accurate methods of nutritional assessment are vital for people suffering from this condition. While assessment of nutritional status in children with CP has been extensively studied, very few studies have been carried out on adults. These limitations are due to the great anatomical-functional variability characteristic of this syndrome. Difficulties that derive from this variability in adult patients with CP mean that there remains an urgent need for certain standards of nutritional assessment for this population. The objective of this review is to compile the latest trends in nutritional assessment in adults with CP to guide the development of a conceptual framework for future research. Recent Findings With this aim, relevant studies have been identified. The most commonly used technique to evaluate nutritional status is the BMI because of its ease-of-use. However, its well-known limitations fail to adequately estimate the nutritional status in this population, with measurements of patients with CP yielding results that are much less accurate than those that already exist in the general population. Summary Although more studies are needed, kinanthropometry is considered one of the most reliable techniques; nevertheless, the anatomical limitation characteristic of CP plays a limiting factor.