Oxidative refolding by Copper-catalyzed air oxidation consistently increases the homogeneity and activity of a Novel Interleukin-2 mutein.

Interleukin-2 (IL-2) has been used in cancer treatment for over 30 years. However, due to its high toxicity, new mutant variants have been developed. These variants retain some of the biological properties of the original molecule but offer other therapeutic advantages. At the Center of Molecular Immunology, the IL-2 no-alpha mutein, an IL-2 agonist with lower toxicity than wtIL-2, has been designed, produced, and is currently being evaluated in a Phase I/II clinical trial. The mutein is produced in E. coli as an insoluble material that must be refolded in vitro to yield a fully active protein. Controlled oxidation steps are essential in the purification process of recombinant proteins produced in E. coli to ensure the proper formation of the disulfide bonds in the molecules. In this case, the new purification process includes a copper-catalyzed air oxidation step to induce disulfide bond establishment. The optimal conditions of pH, copper, protein and detergent concentration for this step were determined through screening. The produced protein demonstrated a conserved 3D structure, higher purity, and greater biological activity than the obtained by established process without the oxidation step. Four batches were produced and evaluated, demonstrating the consistency of the new process.

Efficient capture of recombinant SARS-CoV-2 receptor-binding domain (RBD) with citrate-coated magnetic iron oxide nanoparticles.

Several vaccines against COVID-19 use a recombinant SARS-CoV-2 receptor-binding domain (RBD) as antigen, making the purification of this protein a key step in their production. In this work, citrate-coated magnetic iron oxide nanoparticles were evaluated as nano adsorbents in the first step (capture) of the purification of recombinant RBD. The nanoparticles were isolated through coprecipitation and subsequently coated with sodium citrate. The citrate-coated nanoparticles exhibited a diameter of 10 ± 2 nm, a hydrodynamic diameter of 160 ± 3 nm, and contained 1.9 wt% of citrate. The presence of citrate on the nanoparticles' surface was confirmed through FT-IR spectra and thermogravimetric analysis. The crystallite size (10.1 nm) and the lattice parameter (8.3646 Å) were determined by X-ray diffraction. In parallel, RBD-containing supernatant extracted from cell culture was exchanged through ultrafiltration and diafiltration into the adsorption buffer. The magnetic capture was then optimized using different concentrations of nanoparticles in the purified supernatant, and we found 40 mg mL-1 to be optimal. The ideal amount of nanoparticles was assessed by varying the RBD concentration in the supernatant (between 0.113 mg mL-1 and 0.98 mg mL-1), which resulted in good capture yields (between 83 ± 5% and 94 ± 4%). The improvement of RBD purity after desorption was demonstrated by SDS-PAGE and RP-HPLC. Furthermore, the magnetic capture was scaled up 100 times, and the desorption was subjected to chromatographic purifications. The obtained products recognized anti-RBD antibodies and bound the ACE2 receptor, proving their functionality after the developed procedure.

Topical use of hyperoxygenated fatty acids decreases surgical site infection in patients following laparoscopic cholecystectomy. A randomized controlled trial.

BACKGROUND: To date, the topically effect of hyperoxygenated fatty acids (HOFA) on the control of surgical site infection (SSI) is still unclear. OBJECTIVE: To assess the effect of topical application of a HOFA solution on the umbilical trocar site after laparoscopic cholecystectomy on SSI. The occurrence of trocar site incisional hernia (TSIH) was also analyzed. METHODS: A prospective, double-blind, randomized trial was conducted in patients undergoing laparoscopic cholecystectomy, who also presented at least one of the following associated risk factors for SSI and TSIH such as Body Mass Index (BMI) above 30 kg/m2, Diabetes Mellitus (DM), age over 65 years and Chronic Obstructive Pulmonary Disease (COPD). Patients were randomly allocated to topical application of a HOFA solution (HOFA arm) or saline physiological solution (non-HOFA arm) during closure of the umbilical trocar site with a polypropylene mesh. SSI was the primary outcome. TSIH was also assessed as a secondary outcome. RESULTS: 103 patients were included, 51 (49.5%) in the HOFA group and 52 (50.5%) in the non-HOFA group. SSI rate was significantly lower in the HOFA group in comparison with the non-HOFA group (19.6% vs. 3.8%; p = 0.028). TSIH rates were similar in both groups (3.8% vs. 2%). Multivariate analyses showed that only HOFA decreased significantly SSI rate. CONCLUSION: Topical application of a HOFA solution at the umbilical trocar site after laparoscopy cholecystectomy decreased SSI rate.

Association between offensive and defensive playing style variables and ranking position in a national football league.

The aim of this study was to determine the association of playing style and efficacy variables with football success in a professional football league. Match statistics were obtained from 23 football teams competing in the 2017-2018 and 2018-2019 seasons of the Spanish national league(LaLiga). Offensive and defensive playing style and efficacy variables were calculated. Pearson's correlation coefficient tests and principal component analysis (PCA) were applied to establish the influence of these variables on the number of points obtained at the end of the season and on the ranking position. In isolation, the efficacy of completion showed the highest association with ranking points and position. A two-dimension PCA explained 77.8% of the variance in the ranking position. In dimension-1 (58.5%), game initiative and attack building, and in dimension-2 (19.3%), efficacy of defensive containment and a lower rate of long passes were within the variables that explained more variance in the ranking position. Success in football, measured by ranking position at the end of the Spanish national league, was associated with several playing style and efficacy variables. Overall, a dominant game style with high efficacy to finish attacking plays, and an offensive game initiative, are most associated with successful football.

Injury Profile in Women's Football: A Systematic Review and Meta-Analysis.

BACKGROUND: Football is the most popular sport among women; however, little is known about the injury profile in this population. This information would help design tailored injury risk mitigation strategies that may make football safer for women. OBJECTIVE: The aim of this study was to perform a systematic review and meta-analysis of epidemiological data of injuries in women´s football. METHODS: A systematic review following PRISMA guidelines was performed up to January 2020 in PubMed, Web of Science, Sportdiscus and the Cochrane Library databases. Twenty-two studies reporting the incidence of injuries in women football were analysed. Two reviewers independently extracted data (intraclass correlation coefficient [ICC] for inter-reviewer reliability = 0.87) and assessed study quality using the STROBE statement, GRADE approach, Newcastle Ottawa Scale and Downs and Black assessment tools. Studies were combined in pooled analyses (injury incidence and injury proportion) using a Poisson random effects regression model. RESULTS: The overall incidence of injuries in female football players was 6.1 injuries/1000 h of exposure. Match injury incidence (19.2 injuries/1000 h of exposure) was almost six times higher than training injury incidence rate (3.5 injuries/1000 h of exposure). Lower extremity injuries had the highest incidence rates (4.8 injuries/1000 h of exposure). The most common types of injuries were muscle/tendon (1.8 injuries/1000 h of exposure) and joint (non-bone) and ligament (1.5 injuries/1000 h of exposure), which were frequently associated with traumatic incidents. Slight/minimal injuries (1-3 days of time loss) were the most common. The incidence rate of injuries during matches in the top five world ranking leagues was higher than the rest of the leagues (19.3 vs 10.7 injuries/1000 h of exposure, respectively). The weighted injury proportion was 1.1 (95% confidence interval = 0.6-1.7) whereby on average players sustained more than one injury per season. CONCLUSIONS: Female football players are exposed to a substantial risk of sustaining injuries, especially during matches that require the highest level of performance. To markedly reduce overall injury burden, efforts should focus on introducing and evaluating preventative measures that target match specific dynamics to make football players more capable of responding to the challenges that they have to deal with during match play. REGISTRATION: This systematic review was registered in the PROSPERO international prospective register of systematic reviews (ID = CRD42019118152).

Association Between FGF-23 Levels and Risk of Fracture in Women With Systemic Sclerosis.

INTRODUCTION/BACKGROUND: The purpose of this study was to evaluate the association between Fracture Risk Assessment Tool (FRAX) and serum fibroblast grow factor-23 (FGF-23) levels in SSc women patients compared with healthy controls. METHODOLOGY: This cross-sectional study was performed in San Cecilio Hospital, Granada (Spain) from November 2017 to May 2019. Sixty-two women with SSc and 62 age and sex matched healthy controls were included in this study. FGF-23 serum concentration was evaluated by indirect enzyme-linked immunosorbent assay. The FRAX scoring tool was applied using the on-line calculator (www.shef.ac.uk/FRAX). RESULTS: Even though there was no significant difference in FGF-23 levels between SSc women patients and healthy controls (78.2 ± 60.5 vs 80.3 ± 56.3 pg/mL, p = 0.662). FGF-23 levels were positively associated with FRAX index within the study group. CONCLUSIONS: This study shows that FGF-23 status is associated with FRAX index in women with SSc. FGF-23 could be a promising biomarker for detecting risk fracture in SSc women patients.

Association between homocysteine levels and arterial stiffness in women with systemic sclerosis.

OBJECTIVES: The purpose of this study was to evaluate homocysteine (Hcy) serum levels in women with systemic sclerosis (SSc) compared with healthy controls and to examine possible associations between Hcy and markers of arterial stiffness. METHODS: A cross-sectional study was performed at a single hospital between November 2017 and May 2019: 62 women with SSc and 62 age- and sex-matched healthy controls were enrolled. Pulse wave velocity (PWV) was measured non-invasively along the carotid-femoral arterial segment. Serum Hcy was analysed using immunonephelo-metric method. RESULTS: There was a significant difference in Hcy serum levels between SSc female patients and healthy controls (11.9±3.3 vs. 10.3±2.3 µmol/ml, p=0.002). Serum levels of Hcy were positively correlated with PWV (r=0.28, p<0.05), brain natriuretic peptide (BNP) (r=0.36, p<0.05) and disease duration (r=0.38, p<0.05), within the SSc group. In addition, in the linear regression model, higher Hcy concentrations were associated with higher PWV [ß=0.74 95% CI (0.085, 1.394); p=0.027], BNP [ß=0.04 95% CI (0.014, 0.072); p=0.004] and disease duration [ß=0.18 95% CI (0.070, 0.300); p=0.002]. In multiple linear regression model adjusting for covariants, Hcy remained positively related to the PWV [ß=0.033 95% CI (0.003, 0.062); p=0.031]. CONCLUSIONS: Our findings revealed a positive correlation between Hcy serum levels and PWV, which indicates that high levels of Hcy may predispose to the development of vascular stiffness in patients with SSc.

Role of fibroblast growth factor-23 in calcinosis in women with systemic sclerosis.

OBJECTIVE: Systemic sclerosis (SSc) is a complex disorder of unknown etiology. The purpose of this study was to evaluate fibroblast growth factor-23 (FGF-23) serum levels in women with SSc compared with healthy controls and to examine a possible association between FGF-23 serum levels with the presence of calcinosis in SSc patients. METHODS: This cross-sectional study was performed in San Cecilio Hospital, Granada (Spain) from November 2017 to May 2019. Sixty-two women with SSc and 62 age and sex matched healthy controls were included in this study. FGF-23 serum concentration was evaluated by indirect enzyme-linked immunosorbent assay (ELISA). RESULTS: There was no significant difference in FGF-23 levels between SSc patients and healthy controls (78.2 ± 60.5 vs. 80.3 ± 56.3 pg/mL, p= 0.662). Regarding the characteristics of the disease, we found a relationship between the values of FGF-23 and the presence of calcinosis. The levels of FGF-23 are higher in patients suffering from calcinosis (p= 0.028). CONCLUSION: We observed the presence of higher levels of serum FGF-23 in SSc female patients with calcinosis. Therefore, FGF-23 could be a possible therapeutic target for future treatments.

FGF23-Klotho axis in patients with rheumatoid arthritis.

OBJECTIVES: We aimed to compare serum Klotho and fibroblast growth factor-23 (FGF-23) levels between rheumatoid arthritis (RA) patients and healthy controls. Possible association between FGF-23 and soluble Klotho with different characteristic of the disease as well as their potential role as surrogate markers of cardiovascular disease (CVD) were studied. METHODS: Sixty-three patients with RA recruited at Vega-Baja Hospital, Orihuela (Spain) from November 2016 to May 2018 and sixty-five age- and sex-matched healthy controls were included in this study. Serum Klotho and FGF-23 were analysed using ELISA. RESULTS: Patients had higher serum levels of Klotho than healthy controls (p˂0.0001). They were positively associated with the presence of anticitrullinated peptide antibody and rheumatic factor (p<0.05). Klotho serum levels were higher in RA patients treated with biologic agents than in those undergoing conventional therapy (p=0.008). However, no association with carotid intima media thickness was found. Although no significant differences in serum FGF-23 levels between patients and controls were found (p=0.43), FGF-23 levels were positively associated with low-density lipoprotein (LDL-c) level (p<0.05) and smoking (p=0.008) in patients with RA. CONCLUSIONS: The increased serum Klotho levels in RA patients, especially in those undergoing biologic therapy, may indicate a potential implication in the pathogenesis of the disease. Although levels of FGF-23 were related to LDL-c levels, the FGF-23-Klotho axis does not seem to be related to subclinical arteriosclerosis in RA.

Endothelin-1 serum levels in women with Rheumatoid Arthritis.

OBJECTIVE: The purpose of this study was to evaluate serum Endothelin-1(ET-1) levels in women Rheumatoid Arthritis (RA) patients compared with healthy controls, examine possible associations between ET-1 with different characteristic of the disease and investigate possible associations between ET-1 with surrogate markers of cardiovascular disease (CVD). METHODS: This cross-sectional study was performed in Vega-Baja Hospital, Orihuela (Spain) from November 2016 to May 2018. Sixty-three women with RA and sixty-five age and sex healthy controls were included in this study. Serum ET-1 was analyzed using ELISA. RESULTS: Serum levels of ET-1 in RA women patients were higher than those in healthy controls (p ˂0.001). Serum levels of ET-1 were positively associated with N-terminal pro-brain natriuretic peptide (NT-proBNP) (r = 0.27, p < 0.05) and with C-reactive protein (CRP) (r = 0.36, p < 0.05). ET-1 levels in women with RA were higher in smokers. Prednisone use was associated with lower ET-1 levels. No association with carotid intima media thickness was found. CONCLUSIONS: we observed the presence of higher levels of serum ET-1 in RA women patients compared with healthy controls. These increased levels of ET-1 are associated with inflammation and smoking and reduced by prednisone intake.

Clinical relevance of monitoring serum adalimumab levels in axial spondyloarthritis.

Our aim was to assess the relationship between serum adalimumab levels, anti-drug antibodies (ADA) and disease activity in patients with axial spondylarthritis (SpA). We have carried out a single-centre cross-sectional study. adalimumab and ADA levels were analysed with ELISA and correlated with SpA activity using BASDAI and ASDAS scores. Adalimumab cut-off value was calculated to discriminate inactive disease/low disease activity (BASDAI < 4; ASDAS < 2.1) from moderate/high disease activity (BASDAI ≥ 4; ASDAS ≥ 2.1), using a receiver operating characteristic (ROC) curve. Up to January 2016, 51 consecutive patients were included. The median (range) age was 46.6 (18-68) and 47.1% were women. ADA prevalence was 27.5%, with none detected in the 21.6% receiving concomitant disease-modifying antirheumatic drugs (DMARDs) (p = 0.021). Adalimumab level was normal (> 3 mg/l) in 36 patients (70.6%), all without ADA. Fifteen patients (29.4%) had subtherapeutic adalimumab levels (< 3 mg/l), with ADA in 14 (93%). Median adalimumab (mg/l) was significantly higher in patients with inactive disease/low disease activity: BASDAI < 4 vs ≥ 4: 9.5 vs 2.6 (p < 0.01); ASDAS-CRP < 2.1 vs ≥ 2.1: 9.3 vs 0.3 (p < 0.001); ASDAS-ESR < 2.1 vs ≥ 2.1: 9.9 vs 3.0 (p < 0.001), and this finding was consistent with the result of the multivariate model. Patients with inactive disease/low disease activity presented significantly lower ADA levels. The adalimumab level cut-offs and area under the curve (AUC) obtained in the ROC curves were: ASDAS-CRP (< 2.1) 4.6 mg/l (AUC 81.2%; 95% CI 67.5-94.9; p < 0.001); ASDAS-ESR (< 2.1) 7.7 mg/l (AUC 82.4%; 95% CI 69.3-95.5; p < 0.001); BASDAI (< 4) 6.4 mg/l (AUC 73.5%; 95% CI 58.6-88.3; p < 0.01). In conclusion, presence of ADA in axial SpA patients treated with adalimumab was associated with lower serum drug levels. ADA levels were lower and adalimumab levels were higher in patients with inactive disease/low disease activity based on BASDAI and ASDAS indices. Concomitant treatment with MTX reduces de likelihood of finding ADA. Serum adalimumab levels above 4.6 mg/l are recommended to avoid compromising efficacy.

Dynamic Flux Balance Analysis Using DFBAlab.

Bioprocesses are of critical importance in several industries such as the food and pharmaceutical industries. Despite their importance and widespread application, bioprocess models remain rather simplistic and based on unstructured models. These simple models have limitations, making it very difficult to model complex bioprocesses. With dynamic flux balance analysis (DFBA) more comprehensive bioprocess models can be obtained. DFBA simulations are difficult to carry out because they result in dynamic systems with linear programs embedded. Therefore, the use of DFBA as a modeling tool has been limited. With DFBAlab, a MATLAB code that performs efficient and reliable DFBA simulations, the use of DFBA as a modeling tool has become more accessible. Here, we illustrate with an example how to implement bioprocess models in DFBAlab.

A shift from N-glycolyl- to N-acetyl-sialic acid in the GM3 ganglioside impairs tumor development in mouse lymphocytic leukemia cells.

Humans, in contrast to other mammals, do not synthesize N-glycolyl-neuraminic acid (Neu5Gc) due to a deletion in the gene (cmah) encoding the enzyme responsible for this conversion, the cytidine monophospho-N-acetyl-neuraminic acid hydroxylase (CMP-Neu5Ac hydroxylase). The detection of considerable amounts of Neu5Gc-sialoconjugates, in particular gangliosides, in human malignancies makes these antigens attractive targets for immunotherapy, in particular with monoclonal antibodies (mAbs). We have previously described a GM3(Neu5Gc) ganglioside-specific mAb, named 14F7, with the ability to kill tumor cells in a complement-independent manner. Silencing the cmah gene in GM3(Neu5Gc)-expressing L1210 mouse lymphocytic leukemia B cells caused the abrogation of this cytotoxic effect. We now show that cmah-silenced L1210 cells (cmah-kd) express a high level of GM3(Neu5Ac) and have an impaired ability for anchorage-independent cell growth and tumor development in vivo. No evidences of increased immunogenicity of the cmah-kd cell line were found. These results provide new evidences on the role of GM3(Neu5Gc), or Neu5Gc-sialoconjugates in general, in tumor biology. As an important tool in this study, we used the humanized version (here referred to as 7C1 mAb) of a recently described, rationally-designed mutant of 14F7 mAb that is able to bind to both GM3(Neu5Gc) and GM3(Neu5Ac). In contrast to its parental antibody, the humanized 14F7 (14F7hT) mAb, 7C1 mAb was able to kill not only GM3(Neu5Gc)-expressing L1210 wild type cells, but also GM3(Neu5Ac)-expressing cmah-kd cells, which endorses this antibody as a potential agent for cancer immunotherapy.

[Analysis of an outbreak of viral meningitis in the province of Tucuman, Argentina]. / Análisis de un brote de meningitis viral en la provincia de Tucumán, Argentina.

OBJECTIVE: To confirm the occurrence of an outbreak of viral meningitis in 1996 in the province of Tucuman, Argentina, and to study the outbreak's epidemiological characteristics. METHODS: We analyzed information from the National Epidemiological Surveillance System of the Ministry of Health (MOH) of Argentina for 1994-1998 that had been provided by the MOH's Bureau of Epidemiology. We calculated incidence rates using population estimates for the years 1994-1998 developed by the National Statistics and Census Institute, based on the 1991 census. We studied frequencies with contingency tables, using the chi-square method with Yates' correction. Results were considered significant when P < 0.05. RESULTS: We confirmed the occurrence of an outbreak of 189 cases of viral meningitis between 11 February and 18 May 1996. The incidence of cases in Tucuman province increased between 1995 and 1996, from 0.5 to 19.3 cases per 100 000 person-years. That 1996 rate in Tucuman was significantly higher than what was seen in the rest of the country (2.8 cases per 100 000 person-years). Of the 189 cases, 142 of them (75.1%) occurred in children less than 9 years old. Out of 111 samples studied, 65 of them (58.6%) were enterovirus-positive. Through reverse transcription-nested polymerase chain reaction, enteroviruses were found in 66.3% (53/80) of the cases studied by this method, versus in only 29.6% (24/81) of the cases studied through viral isolation. In the 22 samples that were serotyped, echovirus type 4 was identified in 15 of them (68%): 5 by isolation, 3 by sequencing, and 7 by both methods. During the Tucuman outbreak, at least 56% of the cases were hospitalized. This viral meningitis outbreak shows the capacity of enteroviruses to spread and cause disease. CONCLUSIONS: The use of molecular methods makes it possible to rapidly diagnose the etiological virus and to better control an outbreak. Recognizing this outbreak in Tucuman sooner could have averted the majority of the hospitalizations and the indiscriminate use of antibiotics.

Análisis de un brote de meningitis viral en la provincia de Tucumán, Argentina / Analysis of an outbreak of viral meningitis in the province of Tucuman, Argentina

OBJETIVO: Confirmar la existencia de un brote de meningitis viral en 1996 en la provincia de Tucumán, Argentina, y estudiar sus características epidemiológicas. MÉTODOS: Se analizó información obtenida del Sistema Nacional de Vigilancia Epidemiológica (SINAVE) del Ministerio de Salud de Argentina para el período de 1994-1998, la cual fue provista por la Dirección de Epidemiología de dicho ministerio. Para el cálculo de incidencias se usaron estimaciones poblacionales para los años 1994-1998 realizadas por el Instituto Nacional de Estadística y Censos (INDEC) sobre la base del censo de 1991. El estudio de frecuencias se realizó mediante el análisis de tablas de contingencia de doble entrada, según el método de ji cuadrado con la corrección de Yates. Se consideró significativo el resultado cuando P < 0,05. RESULTADOS: Se confirmó la presencia de un brote de 189 casos entre el 11 de febrero y el 18 de mayo de 1996. La incidencia de casos en la provincia mostró un aumento entre 1995 y 1996 (de 0,5 a 19,3 casos por 100000 años-persona) y dicha incidencia fue significativamenrte mayor que la observada en el resto del país (19,3 frente a 2,8 casos por 100000 años-persona). El 75,1 por ciento de los casos ocurrió en niños menores de 9 años (142/189). Se detectó la presencia de Enterovirus (EV) en 65 de las 111 muestras estudiadas (58,6 por ciento). Mediante la reacción en cadena de la polimerasa (RCP) anidada con transcripción inversa se logró detectar EV en 66,3 por ciento (53/80) de los casos estudiados por este método, en comparación con solo 29,6 por ciento (24/81) de los estudiados mediante aislamiento viral. Se identificó echovirus tipo 4 en 15 (68 por ciento) en las 22 muestras tipificadas (5 por aislamiento, 3 por secuenciación y 7 por ambos métodos). Este brote demuestra la capacidad de los EV para diseminarse y producir enfermedad en la población. Durante el brote, por lo menos 56 por ciento de los casos fueron hospitalizados. CONCLUSIONES: El uso de métodos moleculares permitió el diagnóstico rápido del virus etiológico y posibilitó un mejor control del brote. El reconocimiento temprano de este podría haber evitado la mayoría de las hospitalizaciones y el uso indiscriminado de antibióticos

Objective. To confirm the occurrence of an outbreak of viral meningitis in 1996 in the province of Tucuman, Argentina, and to study the outbreak's epidemiological characteristics. Methods. We analyzed information from the National Epidemiological Surveillance System of the Ministry of Health (MOH) of Argentina for 1994­1998 that had been provided by the MOH's Bureau of Epidemiology. We calculated incidence rates using population estimates for the years 1994­1998 developed by the National Statistics and Census Institute, based on the 1991 census. We studied frequencies with contingency tables, using the chi-square method with Yates' correction. Results were considered significant when P < 0.05. Results. We confirmed the occurrence of an outbreak of 189 cases of viral meningitis between 11 February and 18 May 1996. The incidence of cases in Tucuman province increased between 1995 and 1996, from 0.5 to 19.3 cases per 100 000 person-years. That 1996 rate in Tucuman was significantly higher than what was seen in the rest of the country (2.8 cases per 100 000 person-years). Of the 189 cases, 142 of them (75.1%) occurred in children less than 9 years old. Out of 111 samples studied, 65 of them (58.6%) were enterovirus-positive. Through reverse transcription-nested polymerase chain reaction, enteroviruses were found in 66.3% (53/80) of the cases studied by this method, versus in only 29.6% (24/81) of the cases studied through viral isolation. In the 22 samples that were serotyped, echovirus type 4 was identified in 15 of them (68%): 5 by isolation, 3 by sequencing, and 7 by both methods. During the Tucuman outbreak, at least 56% of the cases were hospitalized. This viral meningitis outbreak shows the capacity of enteroviruses to spread and cause disease. Conclusions. The use of molecular methods makes it possible to rapidly diagnose the etiological virus and to better control an outbreak. Recognizing this outbreak in Tucuman sooner could have averted the majority of the hospitalizations and the indiscriminate use of antibiotics.