Cancer cell-selective induction of mitochondrial stress and immunogenic cell death by PT-112 in human prostate cell lines.

PT-112 is a novel immunogenic cell death (ICD)-inducing small molecule currently under Phase 2 clinical development, including in metastatic castration-resistant prostate cancer (mCRPC), an immunologically cold and heterogeneous disease state in need of novel therapeutic approaches. PT-112 has been shown to cause ribosome biogenesis inhibition and organelle stress followed by ICD in cancer cells, culminating in anticancer immunity. In addition, clinical evidence of PT-112-driven immune effects has been observed in patient immunoprofiling. Given the unmet need for immune-based therapies in prostate cancer, along with a Phase I study (NCT#02266745) showing PT-112 activity in mCRPC patients, we investigated PT-112 effects in a panel of human prostate cancer cell lines. PT-112 demonstrated cancer cell selectivity, inhibiting cell growth and leading to cell death in prostate cancer cells without affecting the non-tumorigenic epithelial prostate cell line RWPE-1 at the concentrations tested. PT-112 also caused caspase-3 activation, as well as stress features in mitochondria including ROS generation, compromised membrane integrity, altered respiration, and morphological changes. Moreover, PT-112 induced damage-associated molecular pattern (DAMP) release, the first demonstration of ICD in human cancer cell lines, in addition to autophagy initiation across the panel. Taken together, PT-112 caused selective stress, growth inhibition and death in human prostate cancer cell lines. Our data provide additional insight into mitochondrial stress and ICD in response to PT-112. PT-112 anticancer immunogenicity could have clinical applications and is currently under investigation in a Phase 2 mCRPC study.

The potent contraceptive gestodene exerts insulinotropic effects through its a-ring reduced metabolites with intrinsic estrogen-like activity in pancreatic ß-cells.

PURPOSE: The contraceptive gestodene is a potent synthetic progestin used in several low-dose contraceptive formulations. Clinical studies reported a relationship between long-term use of combined oral contraceptives containing gestodene (GDN) and profound alterations in glucose metabolism in women. The observation that contraceptive synthetic progestins exert hormone-like effects other than their progestational activities, prompted us to investigate whether GDN may induce estrogen-like effects, even though GDN does not interact with estrogen receptors. The aim of this study was to investigate whether GDN affect pancreatic ß-cell activity, directly or through its conversion to other bioactive metabolites. METHODS: The effects of GDN and its two derivatives 3ß,5α-tetrahydro-GDN and 3α,5α-tetrahydro-GDN on insulin 2 (Ins II) and glucokinase (Gk) expression and glucose-stimulated insulin secretion were determined in pancreatic islets from female rats. RESULTS: Gestodene did exert significant effects on islet ß-cells activity. The most striking finding was that 3ß,5α-tetrahydro-GDN and 3α,5α-tetrahydro-GDN had greater stimulatory effects on Ins II and Gk expression than that observed with GDN, consistent with their effects on glucose-stimulated insulin secretion. The effects on gene expression induced by GDN-derivatives were abolished by ICI 182,780 and MPP. In addition, the presence of inhibitors of androgen and progestin-metabolizing enzymes eliminated gene expression induced by GDN. These results indicated that GDN is metabolized to A-ring reduced metabolites with estrogen-like activities and through this mechanism, GDN may affect ß-cell activity. CONCLUSIONS: Altogether, the data suggest that 19-nortestosterone-derived contraceptives such as GDN, possess insulinotropic effects through their conversion into metabolites with intrinsic estrogen-like activity in pancreatic ß-cells.

A large light-mass component of cosmic rays at 10(17)-10(17.5) electronvolts from radio observations.

Cosmic rays are the highest-energy particles found in nature. Measurements of the mass composition of cosmic rays with energies of 10(17)-10(18) electronvolts are essential to understanding whether they have galactic or extragalactic sources. It has also been proposed that the astrophysical neutrino signal comes from accelerators capable of producing cosmic rays of these energies. Cosmic rays initiate air showers--cascades of secondary particles in the atmosphere-and their masses can be inferred from measurements of the atmospheric depth of the shower maximum (Xmax; the depth of the air shower when it contains the most particles) or of the composition of shower particles reaching the ground. Current measurements have either high uncertainty, or a low duty cycle and a high energy threshold. Radio detection of cosmic rays is a rapidly developing technique for determining Xmax (refs 10, 11) with a duty cycle of, in principle, nearly 100 per cent. The radiation is generated by the separation of relativistic electrons and positrons in the geomagnetic field and a negative charge excess in the shower front. Here we report radio measurements of Xmax with a mean uncertainty of 16 grams per square centimetre for air showers initiated by cosmic rays with energies of 10(17)-10(17.5) electronvolts. This high resolution in Xmax enables us to determine the mass spectrum of the cosmic rays: we find a mixed composition, with a light-mass fraction (protons and helium nuclei) of about 80 per cent. Unless, contrary to current expectations, the extragalactic component of cosmic rays contributes substantially to the total flux below 10(17.5) electronvolts, our measurements indicate the existence of an additional galactic component, to account for the light composition that we measured in the 10(17)-10(17.5) electronvolt range.

Sleep Problems Are Related to a Worse Quality of Life and a Greater Non-Motor Symptoms Burden in Parkinson's Disease.

INTRODUCTION: The aim of the present study was to examine the frequency of self-reported sleep problems and their associated factors in a large cohort of PD patients. METHODS: PD patients and controls, recruited from 35 centers of Spain from the COPPADIS cohort were included in this cross-sectional study. Sleep problems were assessed by the Spanish version of the Parkinson's disease Sleep Scale version 1 (PDSS-1). An overall score below 82 or a score below 5 on at least 1 item was defined as sleep problems. RESULTS: The frequency of sleep problems was nearly double in PD patients compared to controls: 65.8% (448/681) vs 33.5% (65/206) (p < 0.0001). Mean total PDSS score was lower in PD patients than controls: 114.9 ± 28.8 vs 132.8 ± 16.3 (p < 0.0001). Quality of life (QoL) was worse in PD patients with sleep problems compared to those without: PDQ-39SI, 19.3 ± 14 vs 13 ± 11.6 (p < 0.0001); EUROHIS-QoL8, 3.7 ± 0.5 vs 3.9 ± 0.5 (p < 0.0001). Non-motor symptoms burden (NMSS; OR = 1.029; 95%CI 1.015-1.043; p < 0.0001) and impulse control behaviors (QUIP-RS; OR = 1.054; 95%CI 1.009-1.101; p = 0.018) were associated with sleep problems after adjustment for age, gender, disease duration, daily equivalent levodopa dose, H&Y, UPDRS-III, UPDRS-IV, PD-CRS, BDI-II, NPI, VAS-Pain, VAFS, FOGQ, and total number of non-antiparkinsonian treatments. CONCLUSION: Sleep problems were frequent in PD patients and were related to both a worse QoL and a greater non-motor symptoms burden in PD. These findings call for increased awareness of sleep problems in PD patients.

A simple method to evaluate groundwater vulnerability in urbanizing agricultural regions.

A methodology to evaluate groundwater vulnerability was developed and tested in a case study in the Central Valleys of the state of Oaxaca, Mexico, a region known for intensive agricultural activities and poor water management policies. An analysis was conducted to create and evaluate scenarios reflecting anthropogenic and natural stressors on groundwater using an analytical hierarchy process (AHP) and geographic information systems. Uncertainty in the vulnerability model was assessed using a Monte Carlo analysis. Five indices (abstraction (Abs), pollution (Po), runoff (Ru), groundwater recharge (Re), and marginalization (Ma)) were selected after an evaluation of the effects of population growth, climatology, hydrogeological features, and social marginalization on access to groundwater. Abstraction, pollution, and recharge rates are the main drivers of groundwater vulnerability, accounting for 87% of the vulnerability. The analysis revealed that the proposed model generates consistent results and contains low uncertainty. It also showed that more than 50% of the region's groundwater is moderately, and the vulnerability has become increasingly with abstraction, reduced recharge, and pollution (the most sensitive indices), indicating that groundwater in the Central Valleys is under great stress. Pollution and abstraction of groundwater resources are expected to rise in the more vulnerable areas, which will increase water crises and reduce access to water in rural communities. The approach and the indicators establish a baseline for the management and protection of water resources in developing countries where high-resolution data are lacking.

Study of the Huntington's disease IT-15 gene in different ethnic groups in Ecuador.

This study aims to establish the current state of the IT-15 (HTT) gene in different Ecuadorian ethnic groups and patients by determining CAG triplet repeats, compared with the ethnicity of individuals. A total of 412 individuals were studied using nested polymerase chain reaction and Sanger sequencing: 75 individuals were indigenous (Kichwas), 211 mestizos, and 65 Afro-Ecuadorians. We included 31 patients who were clinically diagnosed with Huntington's disease (HD) and relatives of the affected patients (n = 30). Moreover, we correlated the presence of HD in Ecuadorian patients with 46 genetic ancestry-informative insertion-deletion polymorphic markers. We found that 77.20% had <28 CAG repetitions, 18.80% had mutable alleles, 2.27% had incomplete penetrance, and 1.70% reflected >39 repetitions. The average of CAG repetitions was 24 ± 3 for indigenous people; 28 ± 2 for mestizos; and 24 ± 3.2 repetitions for the Afro-Ecuadorians. The ancestral component showed that the main ancestry corresponded to Native Americans (0.873) and European ascendants (0.145), Africans were less represented in the evaluated population (0.018). There was a significant difference between the number of CAG repeats in mestizos and indigenous people (P < .01), suggesting that the Ecuadorian mestizo population has a risk factor for the gene mutation.

The anti-estrogenic activity of indole-3-carbinol in neonatal rat osteoblasts is associated with the estrogen receptor antagonist 2-hydroxyestradiol.

PURPOSE: To gain new insight into the roles of cruciferous vegetable-derived bioactive phytochemicals in bone cells, we investigated the effects of indole-3-carbinol (I3C) on cell proliferation and differentiation in estradiol (E2)-exposed calvarial osteoblasts that were obtained from neonatal rats. METHODS: Osteoblast activity was assessed by analyzing cellular DNA, cell-associated osteocalcin (OC) levels and alkaline phosphatase (AP) activity. We also examined [(3)H]-estrone (E1) metabolism and estrogen-agonistic and estrogen-antagonistic activities of 2-hydroxy (OH) E1 and 2-OHE2 and their capacity to displace [(3)H]-E2 at ER binding sites using competition studies. RESULTS: I3C did not affect on cellular DNA, OC levels or AP activity. However, I3C completely inhibited E2-induced increases in cell proliferation and differentiation in neonatal rat osteoblasts. Metabolic studies demonstrated that I3C promoted the conversion of [(3)H]-E1 to 2-OHE1 and 2-OHE2 and those higher rates of conversion (twofold-threefold) were archived when a higher dose of I3C was applied. Proliferation and differentiation studies showed that 2-OHE2 but not 2-OHE1 inhibited E2-induced increases in cell proliferation and differentiation via an ER-mediated mechanism. Likewise, Esr1 was expressed at high level than Esr2. 2-OHE1 showed no activity or affinity for ER. CONCLUSIONS: This study is the first to show that a bioactive compound derived from cruciferous vegetables, I3C, abolishes the E2-mediated stimulation of cell activities including, proliferation and differentiation, in rat osteoblasts and increases the 2-hydroxylation of E1, resulting in the formation of inactive and anti-estrogenic metabolites. These results suggest that in neonatal rat osteoblasts, the anti-estrogenic effect of I3C is mediated by 2-OHE2 through ER-α.

Probing Atmospheric Electric Fields in Thunderstorms through Radio Emission from Cosmic-Ray-Induced Air Showers.

We present measurements of radio emission from cosmic ray air showers that took place during thunderstorms. The intensity and polarization patterns of these air showers are radically different from those measured during fair-weather conditions. With the use of a simple two-layer model for the atmospheric electric field, these patterns can be well reproduced by state-of-the-art simulation codes. This in turn provides a novel way to study atmospheric electric fields.

A Strategic Study about Quality Characteristics in e-Health Systems Based on a Systematic Literature Review.

e-Health Systems quality management is an expensive and hard process that entails performing several tasks such as analysis, evaluation, and quality control. Furthermore, the development of an e-Health System involves great responsibility since people's health and quality of life depend on the system and services offered. The focus of the following study is to identify the gap in Quality Characteristics for e-Health Systems, by detecting not only which are the most studied, but also which are the most used Quality Characteristics these Systems include. A strategic study is driven in this paper by a Systematic Literature Review so as to identify Quality Characteristics in e-Health. Such study makes information and communication technology organizations reflect and act strategically to manage quality in e-Health Systems efficiently and effectively. As a result, this paper proposes the bases of a Quality Model and focuses on a set of Quality Characteristics to enable e-Health Systems quality management. Thus, we can conclude that this paper contributes to implementing knowledge with regard to the mission and view of e-Health (Systems) quality management and helps understand how current researches evaluate quality in e-Health Systems.

Spontaneous hemoperitoneum secondary to retroperitoneal tumor rupture.

OBJECTIVE: To report a case of GIST type retroperitoneal tumor with spontaneous rupture to the abdominal cavity causing acute abdomen secondary to hemoperitoneum. METHODS/RESULTS: We report the case of an 84 year-old man with history of BPH and chronic atrial fibrillation. He presented to the Emergency Department with diffuse abdominal pain, syncope and accompanying vegetative symptoms. Diagnostic work up showed a 19 cm retroperitoneal mass dependent of the left kidney with active bleeding and secondary hemoperitoneum. Left radical nephrectomy was performed with pathology report of gastrointestinal stromal tumor attached to the renal capsule. CONCLUSIONS: Spontaneous hemoperitoneum is a rare entity and it has various etiologies. It is rarely described in retroperitoneal tumors.

Pulse electrodeposition of CuSbS2 thin films: Role of Cu/Sb precursor ratio on the phase formation and its performance as photocathode for hydrogen evolution§.

In this paper, we outline the development of stoichiometric chalcostibite, CuSbS2 thin films, from a single bath by pulse electrodeposition for its application as a photocathode in photoelectrochemical cells (PEC). The Cu/Sb precursor molar ratio of the deposition bath was varied to obtain stoichiometric CuSbS2 thin films. The optimized deposition and dissolution potentials were -0.72 V and -0.1 V vs saturated calomel electrode, respectively. The formation of CuSbS2 was analyzed using different characterization tools. X-ray diffraction and Raman results showed the formation of the pure chalcostibite phase from a precursor bath with molar ratio Cu/Sb = 0.41. The heterostructure CuSbS2/CdS/Pt was tested as a photocathode in the PEC. The energy positions of the conduction and valence bands were estimated from the Mott Schottky plots. The conduction band and valence band offset of CuSbS2/CdS heterojunction were 0.1 eV and 1.04 eV, respectively. The electric field created in the junction reduced the recombination of the electron/hole pairs and improved charge transfer in the interface. The heterostructure CuSbS2/CdS/Pt demonstrated an improved photocurrent density of 3.4 mA cm-2 at 0 V vs reversible hydrogen electrode. The PEC efficiency obtained from the CuSbS2/CdS heterojunction was 0.56 %. Therefore, we demonstrated the feasibility of an inexpensive technique like electrodeposition for the development of an efficient earth-abundant photocathode.

Allergy and acute leukaemia in children with Down syndrome: a population study. Report from the Mexican inter-institutional group for the identification of the causes of childhood leukaemia.

BACKGROUND: Allergies have been described as protective factors against the development of childhood acute leukaemia (AL). Our objective was to investigate the associations between allergy history and the development of AL and acute lymphoblastic leukaemia (ALL) in children with Down syndrome (DS). METHODS: A case-control study was performed in Mexico City. The cases (n=97) were diagnosed at nine public hospitals, and the controls (n=222) were recruited at institutions for children with DS. Odds ratios (OR) were calculated. RESULTS: Asthma was positively associated with AL development (OR=4.18; 95% confidence interval (CI): 1.47-11.87), whereas skin allergies were negatively associated (OR=0.42; 95% CI: 0.20-0.91). CONCLUSION: Our findings suggest that allergies and AL in children with DS share biological and immune mechanisms. To our knowledge, this is the first study reporting associations between allergies and AL in children with DS.

MtDNA mutation in MERRF syndrome causes defective aminoacylation of tRNA(Lys) and premature translation termination.

We have investigated the pathogenetic mechanism of the mitochondrial tRNA(Lys) gene mutation (position 8344) associated with MERRF encephalomyopathy in several mitochondrial DNA (mtDNA)-less cell transformants carrying the mutation and in control cells. A decrease of 50-60% in the specific tRNA(Lys) aminoacylation capacity per cell was found in mutant cells. Furthermore, several lines of evidence reveal that the severe protein synthesis impairment in MERRF mutation-carrying cells is due to premature termination of translation at each or near each lysine codon, with the deficiency of aminoacylated tRNA(Lys) being the most likely cause of this phenomenon.

Two decades of assistive technologies to empower people with disability: a systematic mapping study.

PURPOSE: Information and Communication Technologies have transformed our lives in different social areas, facilitating interpersonal relationships thanks to technological tools. In the specific case of people with disabilities, Assistive Technologies (ATs) break down barriers and increase opportunities to become active members of society with equal opportunities. MATERIALS AND METHODS: This paper presents a systematic mapping study that analyzes the current state-of-the-art of ATs proposed in the literature to support the empowering of people with disability. Specifically, this paper focuses on (1) describing a global vision of the scientific literature published in the last 20 years about ATs in the computer science field and (2) identifying research needs, gaps, and trends. RESULTS: For this purpose, an in-depth analysis of 389 primary studies is presented. The information obtained from the mapping process is also constrained. Concretely, 35 ATs versus 22 disabilities are compared, obtaining striking peaks for some disabilities described in the discussion. CONCLUSIONS: Finally, the findings show that several areas have been covered only lightly, revealing interesting future directions and challenges for junior researchers.

• ATs have the potential to break down barriers for people with disabilities, enabling them to participate more fully in society. This implies a need for rehabilitation programs to incorporate ATs into their strategies to enhance social inclusion.• Given the transformative role of ICT, rehabilitation programs should focus on helping people with disabilities develop the necessary technological skills to utilize ATs effectively.• This work highlights the diversity of ATs and disabilities, suggesting a need for personalized rehabilitation plans that match specific ATs to individual disabilities.• Rehabilitation professionals should be trained to assess and recommend appropriate ATs for each case. Rehabilitation programs should consider incorporating cutting-edge ATs and staying involved in research to contribute to future developments to cover gaps and challenges identified.

[Findings in brainstem auditory evoked potentials in patients under 2 years of age with a history of intraventricular haemorrhage]. / Hallazgos en los potenciales evocados auditivos del tallo cerebral en pacientes menores de 2 años con antecedente de hemorragia intraventricular.

INTRODUCTION: There are few studies that describe the results of auditory pathway assessment in patients with a history of intraventricular haemorrhage (IVH) during the early years of life. Hypoacusis can occur from the earliest stages of IVH. Brainstem auditory evoked potentials (BAEPs) are a useful tool for diagnosing auditory pathway disorders in early childhood. The aim of the present study was to describe the BAEPs findings in patients under 2 years of age with a history of IVH. PATIENTS AND METHODS: We conducted a retrospective observational study in patients under 2 years of age with a history of IVH referred to our hospital for BAEPs over a period of three years. Patients with genetic syndromes associated with hypoacusis were excluded. BAEPs were used to evaluate the presence or absence of any bioelectrical response and latencies of waves I, III and V, as well as of the intervals I-III, III-V and I-V, and also their morphology, amplitude, synchrony and reproducibility. A descriptive analysis was carried out with the calculation of frequencies and percentages. RESULTS: A total of 122 patients were included. Fifty-one per cent of them had a history of Grade I IVH; 42%, Grade II; and 7%, Grades III or IV. A bioelectrical response was obtained in 243 auditory pathways (99.6%). The morphology was found to be altered in 6.2% of the auditory pathways, while amplitudes were decreased in 2.5% of those tested. Latencies for waves I and III were found to be prolonged in 2% and for wave V in 3.6% of patients. The hearing threshold was normal in 64.8%, and 35.2% of cases presented hypoacusis. CONCLUSIONS: The prevalence of hypoacusis was high in the sample analysed. Systematic follow-up using BAEPs is recommended in order to detect and treat problems in the auditory pathway in patients with IVH in a timely manner.

TITLE: Hallazgos en los potenciales evocados auditivos del tallo cerebral en pacientes menores de 2 años con antecedente de hemorragia intraventricular.Introducción. Existen pocos estudios que describan los resultados de la evaluación de la vía auditiva en pacientes con antecedente de hemorragia intraventricular (HIV) durante los primeros años de vida. La hipoacusia puede presentarse desde los grados iniciales de la HIV. Los potenciales evocados auditivos del tallo cerebral (PEATC) son una herramienta útil para diagnosticar alteraciones de la vía auditiva en la infancia temprana. El objetivo del presente estudio fue describir los hallazgos en los PEATC en pacientes menores de 2 años con antecedente de HIV. Pacientes y métodos. Es un estudio observacional retrospectivo realizado en pacientes menores de 2 años con antecedente de HIV enviados a nuestro hospital para la realización de PEATC en un período de tres años. Se excluyó a pacientes con síndromes genéticos asociados a hipoacusia. A través de los PEATC se evaluó la presencia o la ausencia de respuesta bioeléctrica y latencias de las ondas I, III y V, así como de los intervalos I-III, III-V y I-V, además de su morfología, amplitud, sincronía y la replicabilidad. Se realizó un análisis de tipo descriptivo con cálculo de frecuencias y porcentajes. Resultados. Se incluyó a un total de 122 pacientes. El 51% de ellos tenía antecedente de HIV de grado I; el 42%, de grado II; y el 7%, de grados III o IV. La respuesta bioeléctrica se obtuvo en 243 vías auditivas (99,6%). La morfología se encontró alterada en el 6,2% de las vías auditivas, mientras que las amplitudes estuvieron disminuidas en el 2,5% de las evaluadas. Las latencias para las ondas I y III se encontraron prolongadas en el 2%, y, para la onda V, en el 3,6% de los pacientes. El umbral auditivo fue normal en el 64,8%, y el 35,2% de los casos presentó hipoacusia. Conclusiones. La prevalencia de la hipoacusia fue alta en la muestra analizada. Se recomienda realizar el seguimiento mediante PEATC de forma sistemática con la finalidad de detectar y atender oportunamente problemas de la vía auditiva en pacientes con HIV.

Predictive factors of destination at discharge after spinal cord injury.

INTRODUCTION: One of the main goals of the rehabilitation of patients with spinal cord injury (SCI) is the reintegration of the individual to their family, social, and work setting. The objective of this study was to identify the factors that determine the discharge destination after a traumatic spinal cord injury. MATERIAL AND METHODS: We conducted a retrospective descriptive study of 305 patients with SCI who completed the rehabilitation treatment at the spinal injury unit of Hospital Insular de Gran Canaria between 2001 and 2018. RESULTS: During the study period, we observed an increase in the number of patients referred to long-term care centres, from 9.14% between 2001 and 2010 to 18.4% between 2011 and 2018 (P < .01). Of 20 variables that presented a significant association with destination at discharge in the univariate study, 7 presented a significant association in the multivariate study: age (OR: 1.05; 95% CI, 1.02-1.08), living with a partner (OR: 0.26; 95% CI, 0.09-0.76), residing on another island (OR: 3.57; 95% CI, 1.32-9.63), smoking (OR: 3.44; 95% CI, 1.26-9.44), diabetes (OR: 6.51; 95% CI, 1.46-29.02), history of psychiatric disorders (OR: 3.79; 95% CI, 1.31-10.93), and scores on the Spinal Cord Independence Measure-III (SCIM-III) (OR: 0.48; 95% CI, 0.33-0.69). CONCLUSIONS: Our findings identified advanced age, living on the island of Tenerife, not being married, smoking, type 2 diabetes mellitus, history of psychiatric disorders, and low SCIM-III scores as predictive factors of referral to a long-term care centre in patients with traumatic SCI in the Canary Islands.

Community cluster of meningococcal disease by Neisseria meningitidis serogroup C in Andalusia, Spain, March to May 2011.

Between March and May of 2011, a cluster of three fatal cases of meningococcal sepsis occurred in Andalusia, Spain, in a municipality with a population of around 20,000 inhabitants. The cases were in their mid-teens to early thirties and were notified to the epidemiological surveillance system of Andalusia (Sistema de Vigilancia Epidemiológica de Andalucía, SVEA) during a 68-day period from March through May 2011. All three were infected with the same strain of Neisseria meningitidis serogroup C genosubtype VR1:5-1;VR2:10-8. None of the cases had been previously vaccinated against N. meningitidis serogroup C. Antibiotic post-exposure chemoprophylaxis was administered to close contacts of every diagnosed case. Once the cluster was confirmed, the local population was informed through the media about the control measures taken by the health authorities. The vaccination history against N. meningitidis serogroup C of the population under 25 years-old in the municipality was checked. Vaccination was offered to unimmunised individuals younger than 25 years of age and an additional dose of vaccine was offered to those who had been vaccinated between 2000 and 2006 with a vaccination schedule of three doses before the first year of age. No further cases occurred since the beginning of these actions.