Evaluation of different plasmid DNA delivery system for immunization against HER2/neu in a transgenic murine model of mammary carcinoma

Studies of DNA vaccination against HER2/neu showed the effectiveness of immunization protocols in models of transplantable or spontaneous tumors; scarce information, however, has been provided to identify the procedure of DNA administration that more effectively contributes to the activation of immune system against spontaneously arising HER2/neu-positive tumors. We compared the effectiveness of different procedures of DNA vaccine delivery (intradermic injection (ID), gene gun (GG) delivery and intramuscular injection (IM) alone or with electroporation) in a murine transgenic model of mammary carcinoma overexpressing HER2/neu. We highlighted the role of DNA delivery system in the success of DNA vaccination showing that, among the analysed methods, intramuscular injection of the vaccine, particularly when associated to electroporation, elicits a better protection against HER2/neu spontaneous tumor development inducing antibody and cell-mediated immune responsiveness against HER2/neu and a ThI polarization of the immune response

Twin pregnancy of hydatidiform mole and viable fetus: a late diagnosis - case report / Gestação gemelar de mola hidatiforme e feto viável: um diagnóstico tardio - relato de caso

Twin pregnancy with hydatidiform mole and viable fetus is a rare event, evolving, in most cases, to abortion or medical interruption due the associated risks. Prenatal diagnosis allows individualized, rigorous and specialized monitoring, favoring the early identification of complications. We report, based on information obtained through medical record review, interviews with patient and professionals involved and literature review, a case of twin pregnancy with hydatidiform mole and healthy newborn diagnosed at delivery. Although the uncomplicated outcome that was presented suggests the possibility of expectant conduct, it is important to recognize the rarity of the case and the associated risks, both during pregnancy, after uterine emptying and in future pregnancies, and a follow-up after molar pregnancy is essential, which is interrupted approximately half of the patients. Adherence to follow-up is favored by the correct guidance to pregnant women, and prenatal diagnosis is fundamental for ensuring better doctor-patient dialogue. Therefore, it is important to highlight that, in order to properly manage hydatidiform mole pregnancies, due attention should be paid to the early diagnosis

Production and purification of an immunogenic G protein of rabies virus from S2 insect cells

Rabies virus is still a major public health concern in under developed countries of Africa, Asia and Latin America. WHO estimates that almost 55.000 people die from rabies annually. Although Rabies virus strains from dogs still are the main responsible of causing human disease, the increasing case numbers of bat virus strains being transmitted to humans in America raises concerns. The called secondary transmission cycle of rabies involves cats or dogs which are exposed to contaminated bats and then transmit it to humans. Combating this type of transmission requires new strategies for disease control, including new vaccines. Particularly for cats, the current rabies vaccines have demonstrated to cause vaccine-associated sarcoma as a serious side effect. New rabies vaccines using recombinant technology have been proposed using the Rabies Glycoprotein (G) as antigen. The expression of this molecule in S2 insect cells has already proved the ability of this cell line to produce an immunogenic protein. Although this antigen is recognized as the main protein necessary for protection upon immunization, its utilization as a recombinant vaccine has been impaired by difficulties in its production and mainly purification steps. The G protein was expressed as a membrane bound protein in Drosophila melanogaster S2 cells. This membrane G protein was further solubilized using a standardized solution containing detergent and purified in a novel methodology using affinity chromatography resulting in protein recoveries up to 96 %. The membrane protein and the purified G were used for mice immunization, using or not ISCOM adjuvant resulting in good levels of specific antibodies and neutralizing antibodies. When immunized mice were submitted to lethal rabies virus challenge, the preparations showed to confer protection levels similar to that of commercially available veterinary rabies vaccine. A flow path from the production, purification, formulation and immunization against Rabies using a recombinant G protein is presented as a promisor veterinary vaccine candidate

Secondary Structures of MERS-CoV, SARS-CoV, and SARS-CoV-2 Spike Proteins Revealed by Infrared Vibrational Spectroscopy

All coronaviruses are characterized by spike glycoproteins whose S1 subunits contain the receptor binding domain (RBD). The RBD anchors the virus to the host cellular membrane to regulate the virus transmissibility and infectious process. Although the protein/receptor interaction mainly depends on the spike’s conformation, particularly on its S1 unit, their secondary structures are poorly known. In this paper, the S1 conformation was investigated for MERS-CoV, SARS-CoV, and SARS-CoV-2 at serological pH by measuring their Amide I infrared absorption bands. The SARS-CoV-2 S1 secondary structure revealed a strong difference compared to those of MERS-CoV and SARS-CoV, with a significant presence of extended β-sheets. Furthermore, the conformation of the SARS-CoV-2 S1 showed a significant change by moving from serological pH to mild acidic and alkaline pH conditions. Both results suggest the capability of infrared spectroscopy to follow the secondary structure adaptation of the SARS-CoV-2 S1 to different environments

Inhibition of Asaia in adult mosquitoes causes male-specific mortality and diverse transcriptome changes

Mosquitoes can transmit many infectious diseases, such as malaria, dengue, Zika, yellow fever, and lymphatic filariasis. Current mosquito control strategies are failing to reduce the severity of outbreaks that still cause high human morbidity and mortality worldwide. Great expectations have been placed on genetic control methods. Among other methods, genetic modification of the bacteria colonizing different mosquito species and expressing anti-pathogen molecules may represent an innovative tool to combat mosquito-borne diseases. Nevertheless, this emerging approach, known as paratransgenesis, requires a detailed understanding of the mosquito microbiota and an accurate characterization of selected bacteria candidates. The acetic acid bacteria Asaia is a promising candidate for paratransgenic approaches. We have previously reported that Asaia symbionts play a beneficial role in the normal development of Anopheles mosquito larvae, but no study has yet investigated the role(s) of Asaia in adult mosquito biology. Here we report evidence on how treatment with a highly specific anti-Asaia monoclonal antibody impacts the survival and physiology of adult Anopheles stephensi mosquitoes. Our findings offer useful insight on the role of Asaia in several physiological systems of adult mosquitoes, where the influence differs between males and females

A pilot study on brain plasticity of functional connectivity modulated by cognitive training in mild Alzheimer's disease and mild cognitive impairment

Alzheimer's disease (AD) alters the functional connectivity of the default mode network (DMN) but also the topological properties of the functional connectome. Cognitive training (CT) is a tool to slow down AD progression and is likely to impact on functional connectivity. In this pilot study, we aimed at investigating brain functional changes after a period of CT and active control (AC) in a group of 26 subjects with mild AD (mAD), 26 with amnestic mild cognitive impairment (aMCI), and a control group of 29 healthy elderly (HE) people. They all underwent a CT and AC in a counterbalanced order following a crossover design. Resting-state functional MRI and neuropsychological testing were acquired before and after each period. We tested post-CT and post-AC changes of cognitive abilities, of the functional connectivity of the DMN, and of topological network properties derived from graph theory and network-based statistics. Only CT produced functional changes, increasing the functional connectivity of the posterior DMN in all three groups. mAD also showed functional changes in the medial temporal lobe and topological changes in the anterior cingulum, whereas aMCI showed more widespread topological changes involving the frontal lobes, the cerebellum and the thalamus. Our results suggest specific functional connectivity changes after CT for aMCI and mAD

Temporary dense seismic network during the 2016 Central Italy seismic emergency for microzonation studies

In August 2016, a magnitude 6.0 earthquake struck Central Italy, starting a devastating seismic sequence, aggravated by other two events of magnitude 5.9 and 6.5, respectively. After the first mainshock, four Italian institutions installed a dense temporary network of 50 seismic stations in an area of 260 km2. The network was registered in the International Federation of Digital Seismograph Networks with the code 3A and quoted with a Digital Object Identifier ( https://doi.org/10.13127/SD/ku7Xm12Yy9 ). Raw data were converted into the standard binary miniSEED format, and organized in a structured archive. Then, data quality and completeness were checked, and all the relevant information was used for creating the metadata volumes. Finally, the 99 Gb of continuous seismic data and metadata were uploaded into the INGV node of the European Integrated Data Archive repository. Their use was regulated by a Memorandum of Understanding between the institutions. After an embargo period, the data are now available for many different seismological studies.Publishedid 1825T. Sismologia, geofisica e geologia per l'ingegneria sismicaJCR Journa

Gene Duplication in the Sugarcane Genome: A Case Study of Allele Interactions and Evolutionary Patterns in Two Genic Regions

Sugarcane (Saccharum spp.) is highly polyploid and aneuploid. Modern cultivars are derived from hybridization between S. officinarum and S. spontaneum. This combination results in a genome exhibiting variable ploidy among different loci, a huge genome size (~10 Gb) and a high content of repetitive regions. An approach using genomic, transcriptomic, and genetic mapping can improve our knowledge of the behavior of genetics in sugarcane. The hypothetical HP600 and Centromere Protein C (CENP-C) genes from sugarcane were used to elucidate the allelic expression and genomic and genetic behaviors of this complex polyploid. The physically linked side-by-side genes HP600 and CENP-C were found in two different homeologous chromosome groups with ploidies of eight and ten. The first region (Region01) was a Sorghum bicolor ortholog region with all haplotypes of HP600 and CENP-C expressed, but HP600 exhibited an unbalanced haplotype expression. The second region (Region02) was a scrambled sugarcane sequence formed from different noncollinear genes containing partial duplications of HP600 and CENP-C (paralogs). This duplication resulted in a non-expressed HP600 pseudogene and a recombined fusion version of CENP-C and the orthologous gene Sobic.003G299500 with at least two chimeric gene haplotypes expressed. It was also determined that it occurred before Saccharum genus formation and after the separation of sorghum and sugarcane. A linkage map was constructed using markers from nonduplicated Region01 and for the duplication (Region01 and Region02). We compare the physical and linkage maps, demonstrating the possibility of mapping markers located in duplicated regions with markers in nonduplicated region. Our results contribute directly to the improvement of linkage mapping in complex polyploids and improve the integration of physical and genetic data for sugarcane breeding programs. Thus, we describe the complexity involved in sugarcane genetics and genomics and allelic dynamics, which can be useful for understanding complex polyploid genomes