Rapid Isothermal Amplification and Portable Detection System for SARS-CoV-2.

The COVID-19 pandemic provides an urgent example where a gap exists between availability of state-of-the-art diagnostics and current needs. As assay details and primer sequences become widely known, many laboratories could perform diagnostic tests using methods such as RT-PCR or isothermal RT-LAMP amplification. A key advantage of RT-LAMP based approaches compared to RT-PCR is that RT-LAMP is known to be robust in detecting targets from unprocessed samples. In addition, RT-LAMP assays are performed at a constant temperature enabling speed, simplicity, and point-of-use testing. Here, we provide the details of an RT-LAMP isothermal assay for the detection of SARS-CoV-2 virus with performance comparable to currently approved tests using RT-PCR. We characterize the assay by introducing swabs in virus spiked synthetic nasal fluids, moving the swab to viral transport medium (VTM), and using a volume of that VTM for performing the amplification without an RNA extraction kit. The assay has a Limit-of-Detection (LOD) of 50 RNA copies/µL in the VTM solution within 20 minutes, and LOD of 5000 RNA copies/µL in the nasal solution. Additionally, we show the utility of this assay for real-time point-of-use testing by demonstrating detection of SARS-CoV-2 virus in less than 40 minutes using an additively manufactured cartridge and a smartphone-based reader. Finally, we explore the speed and cost advantages by comparing the required resources and workflows with RT-PCR. This work could accelerate the development and availability of SARS-CoV-2 diagnostics by proving alternatives to conventional laboratory benchtop tests.

Inhibition of SHH pathway mechanisms by arsenic trioxide in pediatric medulloblastomas: a comprehensive literature review.

Recent innovations in the genomic understanding of medulloblastomas have provided new ways to explore this highly invasive malignant brain cancer arising from the cerebellum. Among the four different medulloblastoma subgroups described to date, the sonic hedgehog (SHH) genetic pathway is the pathway activated in the tumorigenesis of medulloblastoma. SHH-related medulloblastomas are usually of nodular/desmoplastic histology and frequently occur in children under the age of three, an age group highly susceptible to the acute and long-term effects of treatment. Several new drugs aimed at SHH modulation are currently under development. This review focuses on the role of arsenic trioxide, a drug well established in clinical practice and probably an under-explored agent in medulloblastoma management, in the SHH pathway.

Review: Novel treatment strategies targeting alpha-synuclein in multiple system atrophy as a model of synucleinopathy.

Neurodegenerative disorders with alpha-synuclein (α-syn) accumulation (synucleinopathies) include Parkinson's disease (PD), PD dementia, dementia with Lewy bodies and multiple system atrophy (MSA). Due to the involvement of toxic α-syn aggregates in the molecular origin of these disorders, developing effective therapies targeting α-syn is a priority as a disease-modifying alternative to current symptomatic treatments. Importantly, the clinical and pathological attributes of MSA make this disorder an excellent candidate as a synucleinopathy model for accelerated drug development. Recent therapeutic strategies targeting α-syn in in vivo and in vitro models of MSA, as well as in clinical trials, have been focused on the pathological mechanisms of α-syn synthesis, aggregation, clearance, and/or cell-to-cell propagation of its neurotoxic conformers. Here we summarize the most relevant approaches in this direction, with emphasis on their potential as general synucleinopathy modifiers, and enumerate research areas for potential improvement in MSA drug discovery.

Inhibition of NF- κ B by Dehydroxymethylepoxyquinomicin Suppresses Invasion and Synergistically Potentiates Temozolomide and γ -Radiation Cytotoxicity in Glioblastoma Cells.

Despite advances in neurosurgery and aggressive treatment with temozolomide (TMZ) and radiation, the overall survival of patients with glioblastoma (GBM) remains poor. Vast evidence has indicated that the nuclear factor NF- κ B is constitutively activated in cancer cells, playing key roles in growth and survival. Recently, Dehydroxymethylepoxyquinomicin (DHMEQ) has shown to be a selective NF- κ B inhibitor with antiproliferative properties in GBM. In the present study, the ability of DHMEQ to surmount tumor's invasive nature and therapy resistance were further explored. Corroborating results showed that DHMEQ impaired cell growth in dose- and time-dependent manners with G2/M arrest when compared with control. Clonogenicity was also significantly diminished with increased apoptosis, though necrotic cell death was also observed at comparable levels. Notably, migration and invasion were inhibited accordingly with lowered expression of invasion-related genes. Moreover, concurrent combination with TMZ synergistically inhibited cell growth in all cell lines, as determined by proliferation and caspase-3 activation assays, though in those that express O(6)-methylguanine-DNA methyltransferase, the synergistic effects were schedule dependent. Pretreatment with DHMEQ equally sensitized cells to ionizing radiation. Taken together, our results strengthen the potential usefulness of DHMEQ in future therapeutic strategies for tumors that do not respond to conventional approaches.

Osteosarcoma arising from osteochondroma of the tibia: case report and cytogenetic findings.

Osteochondroma is a cartilage capped benign tumor developing mainly at the juxta-epiphyseal region of long bones. The rate of malignant transformation, mainly into chondrosarcoma, is estimated to be less than 1-3%. Transformation into osteosarcoma is very rare and has been reported only thirteen times. There is little information on treatment and outcome. We report the case of a secondary osteosarcoma arising in the left tibia of a 23-year-old male, 10 years after the initial diagnosis of osteochondroma and after two partial resections. Malignant transformation occurred at the stalk and not at the cartilage cap, as would normally be expected. Chromosome banding analysis revealed the karyotype: 46,XY, t(3;13)(q21;q34) [2]/46,XY [18]. Records from additional cases will help determine the parameters that define these rare secondary bone lesions.

Tetraploidization in Wilms tumor in an infant.

Genetic instability is frequent in human cancer. Unscheduled tetraploidization can trigger cell transformation and tumorigenesis. We made a cytogenetic analysis by Giemsa-trypsin banding of a stage I, biphasic Wilms tumor diagnosed in a 10-month-old male. An evident karyotypic heterogeneity was found. Four different subclones of tumor cells were observed, with DNA content varying from diploid to near-tetraploid complements. The genetic events involved in the acquisition of aneuploidy in Wilms tumor remain unclear. We hypothesize that initial tetraploidization caused aberrant cell division, leading to abnormal chromosomal segregation, cell transformation and tumorigenesis.

Cytogenetic findings in an epithelioid sarcoma with angiomatoid features. A case report.

Epithelioid sarcoma is a rare, aggressive soft tissue tumor of unknown histogenesis showing predominantly epithelioid cytomorphology. We conducted a conventional and molecular cytogenetic study of a 27-year-old male with epithelioid sarcoma with angiomatoid features. Cytogenetic analysis of epithelioid sarcoma metaphase spreads by GTG-banding revealed a diploid chromosome complement with structural and numerical aberrations. Comparative genomic hybridization analysis demonstrated the amplification of 3p24-pter, 4p15.2-p16 and 18q23, while chromosome losses involved 3p13-p14, 3q24-q26.1, 9q21, and 11q21. Fluorescence in situ hybridization assessment showed normal hybridization patterns for the C-MYC and CCND1 loci; CCND1 RNA overexpression was detected by real-time polymerase chain reaction analysis. Genetic evaluation of this rare condition may be useful in determining if epithelioid sarcoma is associated with a distinct genetic background.

Differential expression of E-cadherin gene in human neuroepithelial tumors.

Cadherins are cell-to-cell adhesion molecules that play an important role in the establishment of adherent-type junctions by mediating calcium-dependent cellular interactions. The CDH1 gene encodes the transmembrane glycoprotein E-cadherin which is important in maintaining homophilic cell-cell adhesion in epithelial tissues. E-cadherin interacts with catenin proteins to maintain tissue architecture. Structural defects or loss of expression of E-cadherin have been reported as a common feature in several human cancer types. This study aimed to evaluate the expression of E-cadherin and their correlation with clinical features in microdissected brain tumor samples from 81 patients, divided into 62 astrocytic tumors grades I to IV and 19 medulloblastomas, and from 5 white matter non-neoplasic brain tissue samples. E-cadherin (CDH1) gene expression was analyzed by quantitative real-time polymerase chain reaction. Mann-Whitney, Kruskal-Wallis, Kaplan-Meir, and log-rank tests were performed for statistical analyses. We observed a decrease in expression among pathological grades of neuroepithelial tumors. Non-neoplasic brain tissue showed a higher expression level of CDH1 gene than did neuroepithelial tumors. Expression of E-cadherin gene was higher in astrocytic than embryonal tumors (P = 0.0168). Low-grade malignancy astrocytomas (grades I-II) showed higher CDH1 expression than did high-grade malignancy astrocytomas (grades III-IV) and medulloblastomas (P < 0.0001). Non-neoplasic brain tissue showed a higher expression level of CDH1 gene than grade I malignancy astrocytomas, considered as benign tumors (P = 0.0473). These results suggest that a decrease in E-cadherin gene expression level in high-grade neuroepithelial tumors may be a hallmark of malignancy in dedifferentiated tumors and that it may be possibly correlated with their progression and dissemination.

Towards further understanding of the co-morbidity between attention deficit hyperactivity disorder and bipolar disorder: a MRI study of brain volumes.

BACKGROUND: Although attention deficit hyperactivity disorder (ADHD) and bipolar disorder (BPD) co-occur frequently and represent a particularly morbid clinical form of both disorders, neuroimaging research addressing this co-morbidity is scarce. Our aim was to evaluate the morphometric magnetic resonance imaging (MRI) underpinnings of the co-morbidity of ADHD with BPD, testing the hypothesis that subjects with this co-morbidity would have neuroanatomical correlates of both disorders. METHOD: Morphometric MRI findings were compared between 31 adults with ADHD and BPD and with those of 18 with BPD, 26 with ADHD, and 23 healthy controls. The volumes (cm(3)) of our regions of interest (ROIs) were estimated as a function of ADHD status, BPD status, age, sex, and omnibus brain volume using linear regression models. RESULTS: When BPD was associated with a significantly smaller orbital prefrontal cortex and larger right thalamus, this pattern was found in co-morbid subjects with ADHD plus BPD. Likewise, when ADHD was associated with significantly less neocortical gray matter, less overall frontal lobe and superior prefrontal cortex volumes, a smaller right anterior cingulate cortex and less cerebellar gray matter, so did co-morbid ADHD plus BPD subjects. CONCLUSIONS: Our results support the hypothesis that ADHD and BPD independently contribute to volumetric alterations of selective and distinct brain structures. In the co-morbid state of ADHD plus BPD, the profile of brain volumetric abnormalities consists of structures that are altered in both disorders individually. Attention to co-morbidity is necessary to help clarify the heterogeneous neuroanatomy of both BPD and ADHD.

Differential expression of E-cadherin gene in human neuroepithelial tumors

Cadherins are cell-to-cell adhesion molecules that play an important role in the establishment of adherent-type junctions by mediating calcium-dependent cellular interactions. The CDH1 gene encodes the transmembrane glycoprotein E-cadherin which is important in maintaining homophilic cell-cell adhesion in epithelial tissues. E-cadherin interacts with catenin proteins to maintain tissue architecture. Structural defects or loss of expression of E-cadherin have been reported as a common feature in several human cancer types. This study aimed to evaluate the expression of E-cadherin and their correlation with clinical features in microdissected brain tumor samples from 81 patients, divided into 62 astrocytic tumors grades I to IV and 19 medulloblastomas, and from 5 white matter non-neoplasic brain tissue samples. E-cadherin (CDH1) gene expression was analyzed by quantitative real-time polymerase chain reaction. Mann-Whitney, Kruskal-Wallis, Kaplan-Meir, and log-rank tests were performed for statistical analyses. We observed a decrease in expression among pathological grades of neuroepithelial tumors. Non-neoplasic brain tissue showed a higher expression level of CDH1 gene than did neuroepithelial tumors. Expression of E-cadherin gene was higher in astrocytic than embryonal tumors (P = 0.0168). Low-grade malignancy astrocytomas (grades I-II) showed higher CDH1 expression than did high-grade malignancy astrocytomas (grades III-IV) and medulloblastomas (P < 0.0001). Non-neoplasic brain tissue showed a higher expression level of CDH1 gene than grade I malignancy astrocytomas, considered as benign tumors (P = 0.0473). These results suggest that a decrease in E-cadherin gene expression level in high-grade neuroepithelial tumors may be a hallmark of malignancy in dedifferentiated tumors and that it may be possibly correlated with their progression and dissemination.

Drug resistance and methylation in myelodysplastic syndrome.

Myelodysplastic syndrome is a clonal hematopoietic stem cell disorder that presents a poor survival for patients treated with standard therapies other than stem-cell transplantation. Multi-drug resistance (MDR) to simultaneous drugs used in chemotherapy is a major concern in the treatment of cancer and also in MDS. ATP-binding cassette (ABC) transporters are involved in the main mechanism that confers drug resistance to cells. Increased expression of drug resistance genes, such as MDR1, MRP1 and LRP, is involved with multi-drug resistance in MDS. The expression of these drug efflux transporters acts in synergy with other alterations, such as epigenetic events, increases in multidrug resistance in MDS. Methylation, the main epigenetic mechanism is widely explored in other hematological malignancies; however, in MDS, this mechanism is poorly investigated. Clinical trials evaluated or are under ongoing evaluation of drugs that abrogated ABC transporters action or reversed the abnormal methylation of some genes in MDS. In this report, we explore the data available in the field of drug resistance and methylation both in pediatric and adult MDS.

Drug-resistance in central nervous system tumors: from the traditional cell-resistance model to the genetically driven approaches on therapy.

The advances in the cure rates observed in the oncology field in the past decades were not fully assembled by primary brain tumors. In this heterogeneous group of diseases, resistance to either chemotherapy or radiotherapy still is a major problem to be addressed. Several genetic and epigenetic events may directly influence the response to treatment in these tumors. Throughout recent discoveries, drug resistance in brain tumors was better understood as a final product of different and complexes pathways that interact and modulate cell performance to treatment. The last years experienced a new paradigm in the way brain tumor drug-resistance genes are elected out of the vast human genomic universe. In the former era, models of cell resistance that were documented on solid tumors other than brain were investigated at the central nervous system's counterpart. Nowadays, genomic-based hypothesis generation, supported by modern genetic technique tolls, seem effective in revealing new candidate-genes that might confer the resistance phenotype. Nevertheless, new treatment approaches and novel drugs based on the pharmacogenomic resistance profile, particularly for brain tumors, are just starting to become a reality for clinical purposes.

Air quality data from large cities.

This paper presents an assessment of the air quality for the principal cities in developed and developing countries. Part of the vast and widely dispersed information on air quality that is available at this time on the Internet was compiled, thus making possible a comprehensive evaluation of the tendencies that emerged at the end of the 20th century. Likewise, these values are compared to the air quality thresholds recommended by two international organizations: guideline levels of the World Health Organization (WHO) and limit values of the European Union (EU), in order to determine air quality concentration levels in large cities around the world. The current situation of air quality worldwide indicates that SO(2) maintains a downward tendency throughout the world, with the exception of some Central American and Asian cities. NO(2) maintains levels very close to the WHO guideline value around the world. For particulate matter, it is a major problem in almost all of Asia, exceeding 300 microg/m(3) in many cities. Ozone shows average values that exceed the selected guideline values in all of the analyses demonstrating that it is a global problem. In general, the worldwide trend is to a reduction in the concentrations of pollutants because of the increasingly strong restrictions which local governments and international organizations impose. However, in poor countries and those with low average incomes, concentrations of air pollutants remain high and the trend will be the elevation of their ground levels as they develop, making the problem even worse.

A twin study of alexithymia.

BACKGROUND: Factors contributing to the development of alexithymia and the nature of alexithymia's relation with trait negative and positive affectivity are unclear. In this study, a twin approach was used to examine the degree of genetic and environmental contributions to the different facets of alexithymia, and the nature of their relations to trait negative and positive affectivity. METHOD: Forty-five monozygotic and 32 same-sex dizygotic twin pairs completed the Toronto Alexithymia Scale-20, the Eysenck Personality Inventory, and a zygosity questionnaire. RESULTS: Model fitting analyses indicated that familial influences contributed significantly to all three facets of alexithymia. Parameter estimates and intraclass correlations suggested, though could not confirm, that it was shared environmental factors that contributed to difficulty identifying and communicating emotions (ID and COM), but shared genetic factors that contributed to externally oriented thinking (EOT). Between-twin cross-trait twin analyses revealed strong correlations between ID and neuroticism, and between COM and extraversion, and suggested that it is shared familial influences which account for these associations. CONCLUSIONS: The results of this study indicate that: (a) the different facets of alexithymia are influenced by familial factors; (b) the previously noted associations between ID and COM and trait affectivity are not merely methodological artifacts; and (c) the associations between ID and COM and trait affectivity are influenced by familial factors. The results also suggest that ID and COM are largely influenced by shared environmental factors, but that EOT is influenced by genetic factors.

Análisis de la alta incidencia de partos en la emergencia del Servicio de Obstetricia del Hospital Central de Maracay: junio-agosto 1998 / Analysis of the high incidence of childbirths at the emergency obstetric service of Maracay Central Hospital: june-august 1998

Luego de la descentralización del sector salud del Estado Aragua, se han venido realizando cambios en función de la distribución de los servicios, a pesar del proceso se mantienen servicios con una alta demanda, que dan como resultado una disminución en la calidad de atención, es el caso del Servicio de Emergencia Obstétrica del Hospital Central de Maracay (HCM), que en 1997 atendió mas de a 25.000 pacientes y un total de 12.653 partos, que representan el 45 por ciento de todos los partos atendidos en el Estado. El objetivo de este trabajo fue conocer la procedencia de las pacientes, si fueron referidas o no, resolución del embarazo y evaluación del control prenatal. Se realizaron 472 entrevistas al azar, a pacientes en trabajo de parto o con indicación de interrupción del embarazo por vía alta que fueron atendidas en la Emergencia del Servicio de Obstetricia durante los meses de junio-agosto de 1998. Los resultados revelaron que un 6,4 por ciento de las entrevistadas procedían de fuera de Aragua; de las procedentes del Estado, la mayoría eran municipios sanitarios cercanos. Sólo el 50 por ciento de las entrevistadas portaban una referencia médica escrita, a pesar que el 94,3 por ciento asistió al control prenatal y cerca del 60 por ciento del grupo en estudio, culminó su embarazo con un parto vaginal simple sin complicaciones, que probablemente pudo haberse resuelto en un nivel de atención de menor complejidad, lo que contribuirá a disminuir la alta demanda que presenta este centro asistencial

[Transient pancytopenia induced by parvovirus B19 in a child with hereditary spherocytosis] / Pancitopenia transitória induzida por parvovírus B19.

OBJECTIVE: To describe the occurrence of transient pancytopenia induced by parvovirus B19 infection in a patient with hereditary hemolytic anemia and to discuss the importance of the diagnosis of this pathology.METHODS: Case report of a child whose diagnosis was made by polymerase chain reaction (PCR) and serology, and review of the literature.CLINICAL REPORT: A twelve year-old male patient with hereditary spherocytosis, presenting non-specific symptoms of an infectious syndrome followed by severe and transient pancytopenia, whose diagnosis was a parvovirus B19 infection.CONCLUSION: The diagnosis of parvovirus infection has a particular importance in hematology, especially on some morbid conditions, among them the hereditary hemolytic anemias. PCR is useful because of its rapidness and sensitivity on the specific diagnosis of this disease.

A twin study of individual differences in perceptual asymmetry.

Individual differences in perceptual asymmetry have been associated with individual differences in cognitive abilities, personality characteristics, and psychiatric symptoms, for which between-person variation appears to be genetically influenced. Perceptual asymmetry scores are also associated with direction of handedness, for which between-person variation does not appear to be genetically influenced. To assess whether between-person variation of perceptual asymmetry scores is genetically influenced, we examined asymmetry on a freevision task of face processing, the Chimeric Faces Task (CFT), in a sample of 31 monozygotic (MZ) and 20 same-sex dizygotic (DZ) twin pairs. MZ and DZ within twin-pair resemblances were compared to assess genetic and familial influences on asymmetric hemispheric function. We found that twins within a pair were no more likely to resemble each other than were unrelated individuals. The results suggest that the between-person variation in CFT perceptual asymmetry is not influenced by genes or shared environment.

Avaliaçäo do conhecimento sobre doenças sexualmente transmissíveis (DSTs) e síndrome da imunodeficiência adquirida (AIDS) entre universitários de Ribeiräo Preto/SP / Evaluation of the knowledge on sexually transmitted diseases (STDs) and acquired immunodeficiency syndrome (AIDS) among universitary students of Ribeiräo Preto/SP

Com o objetivo de avaliar o grau de conhecimento sobre DST/AIDS entre universitários de diferentes áreas, submeteu estudantes de uma faculdade de Ribeiräo Preto-SP a questionários com perguntas abertas e fechadas, anônimos após aquiescência. As respostas foram categorizadas como corretas (C), incorretas (I), entendimento incompleto (EI) e prejudicadas (P), sendo a análise realizada por porcentagens. De 1.200 estudantes, 961 (80,80 por cento) participaram do estudo. O número de respostas näo foi harmonioso para os diferentes itens do questionário. A área em que o aluno estava matriculado näo pareceu influenciar nas respostas. Com relaçäo à transmissäo do HIV, em 2.914 respostas obteve-se 65,37 por cento como categoria EI e em 923 (31,68 por cento) como C. Quanto a medidas preventivas contra a AIDS de 1888 respostas, 1.625 (86,07 por cento) como categoria C e 207 (10,96 por cento) como EI. Sobre medidas preventivas contra AIDS, utilizadas pelo aluno, 1.126 (74,42 por cento) como categoria C e 249 (16,46 por cento) e P. Quanto a medidas preventivas contra DSTs, 1.339 (71,11 por cento) como categoria C e 284 (15,80 por cento) obteve-se 1. Sobre medidas preventivas contra DSTs utilizadas pelo aluno, obteve-se 542 (43,92 por cento) como categoria C e 350 (28,36 por cento) como P. Os universitários em sua maioria parecem possuir um conhecimento teórico correto sobre as medidas preventivas contra à AIDS e DST e entendimento incompleto sobre a transmissäo do HIV. Provavelmente existem fatores relacionados à educaçäo ou culturais que impedem aos universitários fazerem uso das medidas preventivas que conhecem. Há necessidade de identificaçao desses fatores para que os programas de difusäo possam atingir os seus objetivos. A educaçäo continuada ainda se mostra necessária.