Antinociceptive Effect of a p-Cymene/ß-Cyclodextrin Inclusion Complex in a Murine Cancer Pain Model: Characterization Aided through a Docking Study.

Pain is one of the most prevalent and difficult to manage symptoms in cancer patients, and conventional drugs present a range of adverse reactions. The development of ß-cyclodextrins (ß-CD) complexes has been used to avoid physicochemical and pharmacological limitations due to the lipophilicity of compounds such as p-Cymene (PC), a monoterpene with antinociceptive effects. Our aim was to obtain, characterize, and measure the effect of the complex of p-cymene and ß-cyclodextrin (PC/ß-CD) in a cancer pain model. Initially, molecular docking was performed to predict the viability of complex formation. Afterward, PC/ß-CD was obtained by slurry complexation, characterized by HPLC and NMR. Finally, PC/ß-CD was tested in a Sarcoma 180 (S180)-induced pain model. Molecular docking indicated that the occurrence of interaction between PC and ß-CD is favorable. PC/ß-CD showed complexation efficiency of 82.61%, and NMR demonstrated PC complexation in the ß-CD cavity. In the S180 cancer pain model, PC/ß-CD significantly reduced the mechanical hyperalgesia, spontaneous nociception, and nociception induced by non-noxious palpation at the doses tested (p < 0.05) when compared to vehicle differently from free PC (p > 0.05). Therefore, the complexation of PC in ß-CD was shown to improve the pharmacological effect of the drug as well as reducing the required dose.

Analysis of CRISPR/Cas Genetic Structure, Spacer Content and Molecular Epidemiology in Brazilian Acinetobacter baumannii Clinical Isolates.

CRISPR/Cas is a molecular mechanism to prevent predatory viruses from invading bacteria via the insertion of small viral sequences (spacers) in its repetitive locus. The nature of spacer incorporation and the viral origins of spacers provide an overview of the genetic evolution of bacteria, their natural viral predators, and the mechanisms that prokaryotes may use to protect themselves, or to acquire mobile genetic elements such as plasmids. Here, we report on the CRISPR/Cas genetic structure, its spacer content, and strain epidemiology through MLST and CRISPR typing in Acinetobacter baumannii, an opportunistic pathogen intimately related to hospital infections and antimicrobial resistance. Results show distinct genetic characteristics, such as polymorphisms specific to ancestor direct repeats, a well-defined degenerate repeat, and a conserved leader sequence, as well as showing most spacers as targeting bacteriophages, and several self-targeting spacers, directed at prophages. There was a particular relationship between CRISPR/Cas and CC113 in the study of Brazilian isolates, and CRISPR-related typing techniques are interesting for subtyping strains with the same MLST profile. We want to emphasize the significance of descriptive genetic research on CRISPR loci, and we argue that spacer or CRISPR typing are helpful for small-scale investigations, preferably in conjunction with other molecular typing techniques such as MLST.

Gait analysis with muscular fibrosis and treatment with Alpinia zerumbet essential oil in immobilized rats.

The analysis of gait in animals is important for understanding movement disorders in various human pathologies, especially those that develop muscle fibrosis. In the search for treatment alternatives for this problem, essential oils have been studied. Among them, research involving the essential oil of Alpinia zerumbet (EOAz) has been shown to promote relaxation and improve muscle function. Therefore, this study aimed to evaluate the effect of EOAz on gait with muscle fibrosis in immobilized rats. 30 rats (Wistar) were divided into five groups of six animals each: control group (without fibrosis and without treatment), immobilization group (with fibrosis and without treatment), and EOAz treatment groups (with fibrosis and with treatment). The animals were immobilized for 15 days with an ankle plantar flexion orthosis. After this period, they were treated with the oil cutaneously for 30 days. The analysis of behavioral tests before treatment indicated a significant increase in the means of the immobilized groups about to with concerning the control. We conclude that EOAz was effective in improving gait after inducing muscle fibrosis in immobilized rats. Studies are needed to assess the oil's effectiveness in the treatment of muscle fibrosis in human pathologies.

Comparative analysis of different PCR-based strategies for HPV detection and genotyping from cervical samples.

BACKGROUND: Human papillomavirus (HPV) is the main cause of cervical cancer. Polymerase chain reaction (PCR)-based techniques are associated with accurate results with respect to HPV detection and genotyping, being able to identify viral DNA at low levels. However, differences in primer design influence their sensibility and specificity, depending on the HPV type assessed. OBJECTIVE: The aim of the study was to comparatively evaluate the effectiveness of three different PCR-based strategies for HPV detection and genotyping from cervical samples. STUDY DESIGN: The procedures were based on different primer design strategies, using MY09/MY11, EntroA, and type specific multiplex PCR primers. RESULTS: Out of 411 samples of cervical scrapings, 45 (10.9%), 50 (12.2%), and 117 (28.5%) were positive for MY09/MY11, EntroA, and multiplex PCR, respectively. For MY09/MY11 positive samples, 36 were negative for EntroA and 23 for multiplex PCR. For EntroA positive samples, 40 were negative for MY09/MY11 and 26 for multiplex PCR. For multiplex PCR positive samples, 96 were negative for MY09/MY11 and 94 for EntroA. MY09/MY11 identified 12 different HPV types, EntroA detected eight types and multiplex PCR detected 11 HPV types. EntroA primers were able to detect HPV in more samples than MY09/MY11, while multiplex PCR, despite the limited targeted HPV types, presented higher sensibility than the other methods. CONCLUSIONS: The three methods presented different advantages and disadvantages, and the present study reinforces the need to use more than one molecular strategy for HPV detection and genotyping, and the development of novel methods which could overcome the limitations of the existing tests.

Molecular characterization and phylogenetic analysis of Chikungunya virus during the 2016 outbreak in Sergipe, northeastern Brazil.

BACKGROUND: Chikungunya (CHIKV) is an arbovirus transmitted mainly by Aedes aegypti females. CHIKV has been highlighted as the pathogen with the greatest impact due to the high morbidity caused by the infection. In 2016, Brazil experienced an outbreak that affected almost 272 000 people. Here, we performed a molecular characterization and phylogenetic analysis of the CHIKV circulating in 2016 in the state of Sergipe, Brazil. METHODS: A partial region of the E1 gene of 16 CHIKV-positive samples from Sergipe State was amplified and sequenced. RESULTS: All sequences belonged to the East-Central-South-African genotype and three point mutations were verified. Two of them were silent mutations and one was a non-synonymous mutation, which changed lysine to threonine at position 211 in the E1 protein. This mutation was present in 81.2% of the sequences, as well as in other five Brazilian sequences from previous studies. This study found that CHIKV strains circulating in Sergipe during the 2016 outbreak belonged to two different haplotypes. CONCLUSIONS: The strains circulating in Sergipe are phylogenetically close to other Brazilian samples circulating in the northeast and southeast of the country, as well as viruses circulating during the same period in Haiti, indicating the rapid spread of these haplotypes.

Structural and functional impacts of E5 genetic variants of human papillomavirus type 31.

Persistent infections caused by high-risk human papillomavirus (HR-HPV) are important, for the development of cervical lesions, but environmental and genetic factors are also related in the process of carcinogenesis. Among the genetic factors, the genetic variants of HR-HPV appear to be related to the risk of persistent infections. Therefore, the present study investigates variants of HPV31 E5 oncogene in cervical scraping samples from Brazilian women to assess their functional and structural effects, in order to identify possible repercussions of these variants on the infectious and carcinogenic process. Our results detected nucleotide changes previously described in the HPV31 E5 oncogene, which may play a critical role in the development of cancer due to its ability to promote cell proliferation and signal transmission. In our study, the interaction percentage of the 31E5 sequence generated by the Immune Epitope Server database and the Analysis Resource (IEDB) allowed us to include possible immunogenic epitopes with the MHC-I and MHC-II molecules, which may represent a possible relationship between protein suppression of the immune system. In the structural analysis of the HPV31 E5 oncoprotein, the N5D, I48 V, P56A, F80I and V64I polymorphisms can be found inserted within transmembrane regions. The P56A mutation has been predicted to be highly stabilizing and, therefore, can cause a change in protein function. Regarding the interaction of the E5 protein from HPV31 with the signaling of NF-kB pathway, we observed that in all variants of the E5 gene from HPV-31, the activity of the NF-kB pathway was increased compared to the prototype. Our study contributes to a more refined design of studies with the E5 gene from HPV31 and provides important data for a better understanding of how variants can be distinguished under their clinical consequences.

High Genotypic Diversity, Putative New Types and Intra-Genotype Variants of Bovine Papillomavirus in Northeast Brazil.

Bovine papillomavirus (BPV) can cause damage to the epithelial and mucosal tissue and currently presents 28 known types. Not all BPV types are associated with the development of cancer in cattle. Studies have shown that variants of human papillomavirus types can present different pathogenic profiles. However, despite the similarity, it is not yet known whether variants of BPV types can also present varying degrees of pathogenicity. Thus, the aim of this study was to evaluate the genetic variability of BPV types and variants isolated in Northeastern Brazil. Samples were obtained from animals with papillomatous lesions. BPV DNA was detected by the amplification of the L1 gene and genotyping was performed by sequencing. Mutations were analyzed in a phylogenetic, structural and functional context. In total, 52 positive samples were obtained and 11 different BPV types were identified in the samples. Ten putative new BPV types were also identified. In addition, several non-synonymous mutations were identified and predicted to alter protein stability, having an impact on immune evasion. The study demonstrated a high genetic diversity of BPV in the region with a large number of mutations identified, serving as a basis for more efficient control measures to be adopted for bovine papillomatosis.

Functional evaluation of human papillomavirus type 31 long control region variants.

Persistent infections by high-risk human papillomavirus (HR-HPV) are a necessary condition, but not sufficient for cervical cancer development. Genetic variants of HR-HPV appear to be related to the risk of persistent infections. The study performed a functional evaluation of variants of the HPV-31 promoter region (LCR). For this, cloning and subcloning of variants HPV-31/UFPE-21 HPV-31/UFPE-89, HPV-31/UFPE-66, E2 gene and prototype HPV-31 were performed. Transfection with different concentrations of E2 was done and the concentration of 25 ng was determined to be ideal for LCR activation. HPV-31/UFPE-21 and HPV-31/UFPE-89 have a greater ability to alter Nluc reporter gene expression levels and HPV-31/UFPE-66 showed decreased levels of gene expression of Nluc reporter gene compared to control. Statistical analysis showed a significant difference between the polymorphic LCR regions and the control (p < 0.0001). A more refined profile of variants of HPV-31 and its importance for the prognosis of cervical lesions begins to be drawn.

Application of an entropy-based computational strategy to identify genomic markers for molecular detection and typing of human papillomavirus.

Human papillomavirus (HPV) is a diverse group of double-stranded DNA viruses that present high tropism for the epithelium and infect keratinocytes. Currently, over 200 viral types have been identified, and almost 40 types preferentially infect the epithelial cells of the genital tract. Infections caused by HPV are the most prevalent viral infections that are sexually transmitted in the world. Given how HPV infection is one of the key factors in the development of cervical cancer, we need to develop more effective diagnostic methods to correctly diagnose patients. The significance of our research is that we have developed and applied a novel computational approach based on entropy to identify phylogenetically informative genomic regions that could be used as markers for the detection and typing of HPV. We have demonstrated that our strategy is capable of finding phylogenetically informative L1 regions to design a primer set that can be used to accurately detect and genotype HPV isolates.

Molecular characterization of Canis familiaris oral papillomavirus 1 identified in naturally infected dogs from Northeast Brazil.

BACKGROUND: Canine papillomavirus (CPV) has 20 described types associated with papillomas or squamous cell carcinoma (SCC). Knowledge about CPV diversity is scarce. Studies on papillomaviruses that infect other hosts show substantial diversity with some types and variants being associated with cancer. HYPOTHESIS/OBJECTIVES: The aim of this study was to assess the genetic variability of the capsid L1 gene of CPV identified in lesions of naturally infected dogs from Brazil. ANIMALS: Six dogs presenting with oral and cutaneous warts from different veterinary clinics in Sergipe state, Northeast Brazil. METHODS AND MATERIALS: Nine skin biopsy samples were collected for histopathological and molecular analyses. Bioinformatics tools were used for genotyping and diversity analysis. Mutations were characterized based on their impact on the L1 protein structure. RESULTS: Sequences of CPV1 were obtained from exophytic papillomas. These sequences had at least five different mutations showing that all sequences were putative CPV1 variants. One CPV1 sequence, obtained from an oral SCC, had a highly destabilizing substitution in the L1 protein which was likely to be associated with changes in protein function. CONCLUSIONS AND CLINICAL IMPORTANCE: Despite the small number of cases analysed and the partial analysis of L1 nucleotide and amino acid sequences, this study has demonstrated diversity in CPV samples from Northeast Brazil. A putative new CPV1 variant associated with oral SCC, with novel protein structure changing mutations, was identified which may be important for understanding papillomavirus pathogenesis.

Comparative transcriptomic analysis of bovine papillomatosis.

BACKGROUND: Bovine papillomavirus (BPV) belongs to the Papillomaviridae family and infects epithelial cells of bovines and closely related animals, causing hyperproliferative lesions known as warts or papillomas, which may regress or progress to form benign or malignant tumors. The virus enters the host cell and interacts with it by altering the regulation of genes that are responsible for controlling the cell cycle, thus triggering lesion formation. It is not yet known which host genes are regulated by viral infection. Therefore, the objective of this study was to make use of next-generation RNA sequencing methods to identify differentially expressed genes associated with BPV infection, which might elucidate possible marker genes that could be used to control the disease. RESULTS: Transcriptome analysis revealed that 1343 genes were differentially regulated (FDR < 0.05). A comparison of gene expression in infected and noninfected cows indicated that 655 genes were significantly upregulated, and 688 genes were significantly downregulated. Most differentially expressed genes were associated with BPV infection pathways, which supports the hypothesis that viral infection was the mechanism associated with this regulation. CONCLUSIONS: This is the first study that focused on a large-scale evaluation of gene expression associated with BPV infection, which is important to identify possible metabolic pathways regulated by host genes for lesion development. In addition, novel targets could be identified in order to find ligands that interact with BPV, with the aim of interrupting the infection cycle.

Swedish Massage: A Systematic Review of its Physical and Psychological Benefits.

Context • Swedish massage is the systematic application of manual pressure and the movement of soft tissue, with rhythmical pressure and stroking to obtain or maintain health. Studies have revealed its many benefits. Objective • The aim of the current study was to determine the results of past studies that evaluated the benefits of Swedish massage for various populations and to highlight its relevance as an alternative medical practice in health promotion and disease prevention. Methods • The research team performed a review of the literature using the key terms massage, relaxation, and benefits of massage. Original case reports and literature reviews of manual therapy published from 2004 to 2014 were included in the current review. The search was conducted using the SciELO, PubMed, and Medline databases. Setting • The study was performed at the Central Library of the Federal University of Sergipe, in São Cristóvão, Sergipe, Brazil. Participants • Participants in the reviewed studies included varied populations, such as nurses in hospitals, infants and older children, pregnant women, older adults, and cancer patients. Results • A total of 4516 articles were identified for consideration through the electronic database searches. After screening, 117 potentially relevant articles were identified for full review, and 11 studies met the inclusion criteria. The studies showed beneficial results in most cases, including improved nasal breathing and cleanliness in children, reduced lower-back pain for nurses, and improved adrenocortical function in infants. Massage is not completely risk free, however, and should be done by a professional very cautiously. Conclusions • Swedish massage has shown beneficial effects for multiple populations and can be used as a therapy. Its use in disease prevention is highly recommended.

Molecular epidemiology of bovine papillomatosis and the identification of a putative new virus type in Brazilian cattle.

Bovine papillomaviruses (BPVs) are a diverse group of double-stranded DNA viruses, of which 12 viral types have been detected and characterized so far. However, there is still a limited understanding of the diversity of BPV. Several putative new BPVs have been detected and some of these have been recently characterized as new viral types. However, only a very limited amount of information is available on the pathology associated with these novel viral types yet this information could be of significant value in improving our understanding of the biology of BPV. The objective of this study was to examine some of the epidemiological features of cutaneous bovine papillomatosis in Brazilian cattle, in particular to establish the relationship between BPV types isolated from beef and dairy cattle herds and the lesions they cause. Seventy-two cutaneous lesions were collected from 60 animals. Histopathological, PCR and sequencing assays were conducted to characterize the lesions and detect the BPV types responsible. Phylogenetic analysis was carried out using the maximum likelihood method. BPV types 1-6 and 8-10 were found, as well as a putative new BPV type that belongs to the Deltapapillomavirus genus. The tumors were all classified as fibropapillomas. This is believed to be the first record of BPV types 3 and 10 associated with fibropapillomas. These results confirm that there is a wide range of BPV types that infect cattle, and that an understanding of this diversity is necessary for improved methods of therapeutic treatment.

An entropy-based approach for the identification of phylogenetically informative genomic regions of Papillomavirus.

The papillomaviruses form a highly diverse group that infect mammals, birds and reptiles. We know little about their genetic diversity and therefore the evolutionary mechanisms that drive the diversity of these viruses. Genomic sequences of papillomaviruses are highly divergent and so it is important to develop methods that select the most phylogenetic informative sites. This study aimed at making use of a novel approach based on entropy to select suitable genomic regions from which to infer the phylogeny of papillomavirus. Comparative genomic analyzes were performed to assess the genetic variability of each gene of Papillomaviridae family members. Regions with low entropy were selected to reconstruct papillomavirus phylogenetic trees based on four different methods. This methodology allowed us to identify regions that are conserved among papillomaviruses that infect different hosts. This is important because, despite the huge variation among all papillomaviruses genomes, we were able to find regions that are clearly shared among them, presenting low complexity levels of information from which phylogenetic predictions can be made. This approach allowed us to obtain robust topologies from relatively small datasets. The results indicate that the entropy approach can successfully select regions of the genome that are good markers from which to infer phylogenetic relationships, using less computational time, making the estimation of large phylogenies more accessible.

New variants of E6 and E7 oncogenes of human papillomavirus type 31 identified in Northeastern Brazil.

OBJECTIVE: We sought to characterize E6 and E7 oncogenes genetic variability of HPV-31 isolated from cervical scraping samples of Northeastern Brazilian women. METHODS: E6 and E7 were amplified with specific primers, cloned and sequenced. The sequences obtained were aligned with the GenBank reference sequences with the aim of evaluating the possible genetic variants. RESULTS: We identified several genetic variants in E6 and E7 sequences from HPV-31 positive women. Three nucleotide changes in E6 were described for the first time in this study. Some nucleotide changes were non-synonymous substitutions. CONCLUSION: The knowledge of region/country HPV specific genetic variations is relevant to understand the epidemiology and the development of effective vaccines.