Influence of two educational Escape Room- Breakout tools in PSTs' affective and cognitive domain in STEM (science and mathematics) courses.

During the last decade, there has been a strong emphasis on developing new instruction methodologies for the effective teaching of different contents. Here, it is important to teach Science, Technology, Engineering and Mathematics (STEM) education, specially, in scientific and mathematical concepts. In the context of active learning and gamification, educational Escape Room - Breakout (ERB) could be a useful strategy to improve students' affective and cognitive domain towards STEM (science and mathematics). Thus, two didactic tools, based on an ERB, have been designed to teach science and mathematics contents. This research compares the influence of two ERBs (Science ERB and Mathematics ERB) in Pre-Service Teachers' (PSTs) affective domain (emotions, attitudes, and self-efficacy towards STEM) and cognitive domain (performance). Non-parametric statistical tests were used, the Mann-Whitney U test was applied to measure significant differences between the variables in the two ERBs. Spearman correlation coefficient was implemented to measure the correlations between the study variables. The results show that there is a significant increase in positive emotions in both ERBs. The emotions "joy", "fun", and "nervousness" are significantly higher after the Science ERB, and the emotion "fear" is lower with respect to the Mathematics ERB. In the self-efficacy and attitudes analysis, a significant increase of 8 items of the questionnaire is observed in the Mathematics ERB with respect to the Science ERB. According to performance analysis, PST grades have been increased after each ERB. Finally, the correlation analysis between variables indicates that positive emotions, high self-efficacy, and positive attitudes increase the PSTs' performance. Here, high values of these variables are related to high values on the theoretical content test after both ERBs. According to these results, the two ERBs used could have several advantages in the PSTs' affective and cognitive domain.

Positive selection pressure on E2 protein of classical swine fever virus drives variations in virulence, pathogenesis and antigenicity: Implication for epidemiological surveillance in endemic areas.

Classical swine fever (CSF), caused by CSF virus (CSFV), is considered one of the most important infectious diseases with devasting consequences for the pig industry. Recent reports describe the emergence of new CSFV strains resulting from the action of positive selection pressure, due mainly to the bottleneck effect generated by ineffective vaccination. Even though a decrease in the genetic diversity of the positively selected CSFV strains has been observed by several research groups, there is little information about the effect of this selective force on the virulence degree, antigenicity and pathogenicity of this type of strains. Hence, the aim of the current study was to determine the effect of the positive selection pressure on these three parameters of CSFV strains, emerged as result of the bottleneck effects induced by improper vaccination in a CSF-endemic area. Moreover, the effect of the positively selected strains on the epidemiological surveillance system was assessed. By the combination of in vitro, in vivo and immunoinformatic approaches, we revealed that the action of the positive selection pressure induces a decrease in virulence and alteration in pathogenicity and antigenicity. However, we also noted that the evolutionary process of CSFV, especially in segregated microenvironments, could contribute to the gain-fitness event, restoring the highly virulent pattern of the circulating strains. Besides, we denoted that the presence of low virulent strains selected by bottleneck effect after inefficient vaccination can lead to a relevant challenge for the epidemiological surveillance of CSF, contributing to under-reports of the disease, favouring the perpetuation of the virus in the field. In this study, B-cell and CTL epitopes on the E2 3D-structure model were also identified. Thus, the current study provides novel and significant insights into variation in virulence, pathogenesis and antigenicity experienced by CSFV strains after the positive selection pressure effect.

Attenuation of disease phenotype through alternative translation initiation in low-penetrance retinoblastoma.

Hereditary predisposition to retinoblastoma (RB) is caused by germline mutations in the retinoblastoma 1 (RB1) gene and transmits as an autosomal dominant trait. In the majority of cases disease develops in greater than 90% of carriers. However, reduced penetrance with a large portion of disease-free carrier is seen in some families. Unambiguous identification of the predisposing mutation in these families is important for accurate risk prediction in relatives and their genetic counseling but also provides conceptual information regarding the relationship between the RB1 genotype and the disease phenotype. In this study we report a novel mutation detected in 10 individuals of an extended family, only three of whom are affected by RB disease. The mutation comprises a 23-basepair (bp) duplication in the first exon of RB1 (c.43_65dup) producing a frameshift in exon 1 and premature chain termination in exon 2. Mutations resulting in premature chain termination classically are associated with high penetrance disease, as message translation may not generate functional product and nonsense mediated RNA decay (NMD) frequently eliminates the mutant transcript. However, appreciable NMD does not follow from the mutation described here and transcript expression in tissue culture cells and translation in vitro reveals that alternative in-frame translation start sites involving Met113 and possibly Met233 are used to generate truncated RB1 products (pRB94 and pRB80), known and suspected to exhibit tumor suppressor activity. These results strongly suggest that modulation of disease penetrance in this family is achieved by internal translation initiation. Our observations provide the first example for rescue of a chain-terminating mutation in RB1 through alternative translation initiation.

A two base pair deletion in the PQBP1 gene is associated with microphthalmia, microcephaly, and mental retardation.

X-linked mental retardation has been traditionally divided into syndromic (S-XLMR) and non-syndromic forms (NS-XLMR), although the borderlines between these phenotypes begin to vanish and mutations in a single gene, for example PQBP1, can cause S-XLMR as well as NS-XLMR. Here, we report two maternal cousins with an apparently X-linked phenotype of mental retardation (MR), microphthalmia, choroid coloboma, microcephaly, renal hypoplasia, and spastic paraplegia. By multipoint linkage analysis with markers spanning the entire X-chromosome we mapped the disease locus to a 28-Mb interval between Xp11.4 and Xq12, including the BCOR gene. A missense mutation in BCOR was described in a family with Lenz microphthalmia syndrome, a phenotype showing substantial overlapping features with that described in the two cousins. However, no mutation in the BCOR gene was found in both patients. Subsequent mutation analysis of PQBP1, located within the delineated linkage interval in Xp11.23, revealed a 2-bp deletion, c.461_462delAG, that cosegregated with the disease. Notably, the same mutation is associated with the Hamel cerebropalatocardiac syndrome, another form of S-XLMR. Haplotype analysis suggests a germline mosaicism of the 2-bp deletion in the maternal grandmother of both affected individuals. In summary, our findings demonstrate for the first time that mutations in PQBP1 are associated with an S-XLMR phenotype including microphthalmia, thereby further extending the clinical spectrum of phenotypes associated with PQBP1 mutations.

A subtelomeric translocation apparently implied in multiple abortions.

PURPOSE: Chromosomal alterations are a major cause of clinically recognized abortions. So cryptic chromosomal rearrangements could be responsible for recurrent miscarriages. METHODS: The study was performed on 18 clinically normal couples who had four or more spontaneous abortions and whose karyotypes were found to be normal. Fluorescence in situ hybridization with specific probes of subtelomeric regions was used after optimizing the amount of probe. RESULTS: Only one subtelomeric translocation was detected between 2p and 3p chromosomes, in a female with seven miscarriages. CONCLUSION: Subtelomeric rearrangements are infrequently related to multiple miscarriages. This kind of studies can be considered specially worthy when a couple have had not only abortions but also offspring with mental and physical anomalies. Thus, possible initial studies should focus on affected children or fetus.

Molecular detection of Spanish deltabeta-thalassemia associated with beta-thalassemia identified during prenatal diagnosis.

BACKGROUND: beta-Thalassemia is one of the most common single gene disorders and is widely distributed in the Mediterranean region. The deltabeta-thalassemias are a rare group of disorders, however in Spain the Spanish deltabeta0 thalassemia that removes 114 kb of beta-globin cluster is quite frequent. METHODS: To establish a molecular diagnosis of beta-thalassemia in a second-trimester fetus from a couple who are beta-thalassemia carriers, a rapid real-time PCR method was performed to detect major mutations prevalent in people of the Mediterranean basin (IVS I-1, IVS I-6, IVS I-110, CD-6, CD-37, and CD-39). The Spanish deltabeta0 deletion was detected using PCR with deletion-specific primers. RESULTS: The father was diagnosed as a carrier for the IVSI-6 (T>C) mutation and the mother as carrier for the Spanish deltabeta0 deletion. The fetus was a compound heterozygote for both those mutations. The phenotype of compound heterozygosity for deltabeta/beta-thalassemia can range from mild thalassemia intermedia to thalassemia major. After confirming these results in the DNA from amniocentesis obtained at the 20th week of gestation, the couple decided to terminate the pregnancy. CONCLUSIONS: This paper reports the first molecular characterization of Spanishdeltabeta0-thalassemia associated with beta-thalassemia IVSI-6, and confirms the importance of prenatal genetic testing for single gene disorder such as beta-thalassemia.

Localization of MRX82: a new nonsyndromic X-linked mental retardation locus to Xq24-q25 in a Basque family.

Clinical and molecular studies are reported on a Basque family (MRX82) with nonsyndromic X-linked mental retardation (XLMR) in five affected males. A total of 38 microsatellite markers were typed. The XLMR locus has been linked to DXS8067, DXS1001, DXS425, DXS7877, and DXS1183 with a maximum LOD score of 2.4. The haplotype studies and multipoint linkage analysis suggest a localization of the MRX82 locus to an interval of 7.6 Mb defined by markers DXS6805 and DXS7346, in Xq24 and Xq25, respectively. No gene contained in this interval has been so far associated with nonsyndromic mental retardation, except for GRIA3, disrupted by a balanced translocation in a female patient with bipolar affective disorder and mental retardation. However, the search for mutations of this gene did not showed a pathogenic mutation in the present family. Given that there are other eight MRX families overlapping this interval, none of them with known mutation, we conclude that at least one new gene responsible for nonsyndromic mental retardation is located in this region.

Auditory neuropathy in patients carrying mutations in the otoferlin gene (OTOF).

Inherited hearing impairment affects one in 2,000 newborns. Nonsyndromic prelingual forms are inherited mainly as autosomal recessive traits, for which 16 genes are currently known. Mutations in the genes encoding connexins 26 and 30 account for up to 50% of these cases. However, the individual contribution of the remaining genes to the whole remains undetermined. In addition, for most of the genes there is a need for studies on genotype-phenotype correlations, to identify distinctive clinical features which may direct the molecular diagnosis to specific genes. Here we present a mutation analysis and a genotype-phenotype correlation study on the gene encoding otoferlin (OTOF), responsible for the DFNB9 subtype of prelingual hearing impairment. Four novel mutations were identified: c.2122C>T (p.Arg708Ter), c.4275G>A (p.Trp1425Ter), c.4362+2T>G, and c.5860_5862delATC (p.Ile1954del). A total of 37 subjects with mutations in OTOF were studied clinically. They were phenotypically homogeneous, having profound hearing impairment with very early onset, as shown by pure-tone audiometry and auditory brainstem responses. Magnetic resonance imaging and computed tomography did not reveal any inner ear malformation. Unexpectedly, transient evoked otoacoustic emissions (TEOAEs) were present, either bilaterally or unilaterally in 11 subjects. Altogether, clinical data of these subjects met the diagnostic criteria of auditory neuropathy. A total of 10 subjects had been successfully provided with cochlear implants. The results of our study indicate that genetic diagnosis of subjects with auditory neuropathy and profound hearing impairment should be directed to the otoferlin gene. Our data are of concern to universal screening programs which use TEOAEs as the first detection test for hearing impairment in newborns, since this technique may overlook a nonnegligible proportion of cases.

Identification and characterization of a novel splice-site mutation in a patient with Wiskott-Aldrich syndrome.

Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder characterized by immunodeficiency, eczema, and thrombocytopenia with small platelets. A wide spectrum of mutations in the Wiskott-Aldrich syndrome protein ( WASP) gene have been identified as causative of the disease. In the present paper, we report on a family with a boy affected by WAS, with a splice-site mutation caused by a T to G substitution in the +2 position of intron 6 (IVS6+2T>G). Expression studies performed in COS-7 and U-937 cells showed that the mutation affected the normal splicing process. As a consequence, an abnormally long transcript of 38 nucleotides is generated. Such missplicing is probably due to the activation of a cryptic splice donor site located 38 nt downstream of exon 6. The translation of such aberrant mRNA will produce a truncated protein with a premature stop at codon 190. Thus, a novel splice-site mutation is reported in a patient with a mild WAS phenotype.

Diagnóstico temprano del retinoblastoma: importancia de la búsqueda de mutaciones en el gen RB1 / Early diagnosis of retinoblastoma: usefulness of searching for RB1 gene mutations

Fundamento: El retinoblastoma, cáncer intraocular infantil más frecuente, se presenta como esporádico (unilateral o bilateral) o afectando a varios miembros de una familia. En los casos hereditarios aparece por una mutación germinal transmitida o surgida de novo, mientras que en los no hereditarios se debe a dos mutaciones somáticas en una célula de la retina. El presente trabajo se planteó con objeto de analizar desde el punto de vista genético el gran número de familias con algún miembro afectado de retinoblastoma, recopiladas en los últimos años, para ahondar en los mecanismos moleculares que inciden en este proceso patológico y ofrecerles consejo genético. Pacientes y método: Se han analizado 59 familias con algún miembro afectado de retinoblastoma y se ha realizado un estudio citogenético, con marcadores polimórficos y de búsqueda de mutaciones en el ADN de leucocitos y de los tumores disponibles. Resultados: En 4 de los 5 casos familiares, se ha establecido la mutación asociada a la enfermedad, al igual que en 9 de los 13 casos bilaterales esporádicos. En un 7 por ciento de los casos unilaterales esporádicos la mutación se encontraba en el ADN leucocitario. La pérdida de heterocigosidad como segundo episodio mutacional se debe principalmente a la recombinación mitótica. Conclusiones: Entre las mutaciones contabilizadas en nuestra serie, se observa mayor frecuencia de mutaciones puntuales de carácter constitucional, que dan origen al primer acontecimiento mutacional, mientras que la pérdida de heterocigosidad es el episodio mutacional detectado mayoritariamente en tumores (AU)