Análise termográfica da ação do jato de plasma no tratamento de rugas faciais ­ série de casos clínicos / Thermographic analysis of plasma jet action in the treatment of facial wrinkles ­ serie of clinical cases

Introdução: o processo de senescência do indivíduo ocorre de forma lenta e contínua e pode gerar inúmeras alterações, dentre elas o envelhecimento cutâneo, derivado do declínio das atividades celulares. Inúmeros recursos têm sido criados ao longo dos anos com a finalidade de frear e ou reverter os aspectos inestéticos da pele, ocasionados por esse processo. Um novo recurso terapêutico vem sendo usado com o objetivo de promover o rejuvenescimento através da diminuição de rugas e linhas de expressão, trata-se do Jato de plasma. Objetivo: avaliar o padrão de variação térmica do tecido tratado com jato de plasma, através da termografia infravermelha. Metodologia: trata-se de uma série de casos clínicos que envolveu 5 pacientes mulheres, com idade acima de 40 anos que apresentavam rugas faciais. As pacientes foram avaliadas e anestesiadas previamente. Em seguida, foi realizada a terapia com o jato de plasma, na região supraorbital. As pacientes foram avaliadas termograficamente antes e depois da anestesia e pósterapia. Resultado: constatou-se através da análise termográfica, uma significativa variação no coeficiente de temperatura da pele onde foi aplicado o anestésico e em seguida o jato de plasma (ΔT > 0,4 °C), em todas as pacientes. Conclusão: o presente estudo comprovou através da utilização da termografia, que o jato de plasma foi capaz de gerar um aumento da temperatura local. Os autores sugerem que tal variação térmica pode ser resultante de um processo de vasodilatação na região tratada.

Introduction: individual's senescence process takes place slowly and continuously beyond generates several changes including skin aging, since the decline in cellular activities. Countless resources have been created over the years, with the goal of stopping and / or reversing the unsightly aspects of the skin caused by this process. Thus, plasma jet, a new therapeutic resource has been used in order to promote rejuvenation through the reduction of wrinkles and expression lines. Objective: To evaluate the pattern of thermal variation of the tissue treated with a plasma jet, using infrared thermography. Methodology: this was a series of clinical cases involving 5 female patients over the age of 40 who had facial wrinkles. Patients were previously evaluated and anesthetized. Then, plasma jet therapy was performed in the supra-orbital region. Patients were evaluated for thermal imaging before and after anesthesia and post-therapy. Result: it was found through thermographic analysis, a significant variation in the temperature coefficient of the skin of all patients where the anesthetic and plasma jet were applied (ΔT > 0.4 °C). Conclusion: the present study proved through the use of thermography that the plasma jet was able to generate an increase in local temperature. The authors suggest that such thermal variation may be the result of a process of vasodilation in the treated region.

Non-pharmacologicalinterventions in the control of urinary incontinence and the instruments that allow the diagnosis: integrative review / Intervenções não farmacológicas no controle da incontinência urinária e os instrumentos que permitam o seu diagnóstico: revisão integrativa

Introduction: Urinary incontinence is a major health problem resulting in physical, psychological and social changes with economic repercussions on the health system. Is a multifactorial condition associated with age-related changes and disorders of the genitourinary system, which corroborates the fact that it is the most often recurring geriatric syndrome. Aims: To identify non-pharmacological interventions for adults with urinary incontinence and to identify tools for urinary incontinence diagnosis in adults. Method: An integrative review study design was completed. Two electronic databases was search (MEDLINE and Web of Science). Three independent reviewers searched databases according to a predetermine inclusion and exclusion criteria. Results: Twelve articles were included in the review. Eleven articles mentioned non-pharmacological interventions, such as physical therapies, lifestyle strategies, behavioural therapies and alternative conservative management options. These interventions should be targeted and individualized to the type of incontinence to result in health gains for the population. One article mentioned an assessment tool for urinary incontinence - The Gaudenz-Fragebogen tool. The evaluation tools can help to systematize the diagnostic activity and consequently improve the clinical practice in the field of urinary incontinence. Conclusion: In care conception, nurses should target their interventions to personal data to address individual symptoms and use assessment tools that can help in the differential diagnosis of UI. Then, to advancing the quality and rigor of nursing care, we advocate that providing nurses with skills in attaining a differential diagnosis of UI presents an added value to the improvement of quality of care in a multidisciplinary context

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Efficient Production of Papillomavirus Gene Delivery Vectors in Defined In Vitro Reactions.

Papillomavirus capsids can package a wide variety of nonviral DNA plasmids and deliver the packaged genetic material to cells, making them attractive candidates for targeted gene delivery vehicles. However, the papillomavirus vectors generated by current methods are unlikely to be suitable for clinical applications. We have developed a chemically defined, cell-free, papillomavirus-based vector production system that allows the incorporation of purified plasmid DNA (pseudogenome) into high-titer papillomavirus L1/L2 capsids. We investigated the incorporation of several DNA forms into a variety of different papillomavirus types, including human and animal types. Our results show that papillomavirus capsids can package and transduce linear or circular DNA under defined conditions. Packaging and transduction efficiencies were surprisingly variable across capsid types, DNA forms, and assembly reaction conditions. The pseudoviruses produced by these methods are sensitive to the same entry inhibitors as cell-derived pseudovirions, including neutralizing antibodies and heparin. The papillomavirus vector production systems developed in this study generated as high as 1011 infectious units/mg of L1. The pseudoviruses were infectious both in vitro and in vivo and should be compatible with good manufacturing practice (GMP) requirements.

Papillomavirus assembly: An overview and perspectives.

Papillomavirus life cycle is tightly coupled to epithelial cell differentiation, which has hindered the investigation of many aspects of papillomavirus biology, including virion assembly. The development of in vitro production methods of papillomavirus pseudoviruses, and the production of "native" virus in raft cultures have facilitated the study of some aspects of the assembly process. In this paper we review the current knowledge of papillomavirus assembly, directions for future research, and the implications of these studies on new therapeutic interventions.

Patterns of Response in Parents of Children With Cancer: An Integrative Review.

PROBLEM IDENTIFICATION: To identify patterns of response of parents in relation to taking care of their child with cancer.
. LITERATURE SEARCH: The search was performed using CINAHL® and Scopus in February 2013.
. DATA EVALUATION: The selection process resulted in 18 articles with a wide range of methodologic approaches. The description of the research methods of each study and the relevance of the results in comparison to the purpose of this review were established as assessment criteria.
. SYNTHESIS: The results of the studies were analyzed using Meleis's Transition Theory, identifying a vast number of patterns of response developed by the parents. These patterns of response were analyzed, compared, and split into four themes.
. CONCLUSIONS: Using this methodology, a wide range of behaviors, attitudes, and competencies associated with the circumstance of parents caring for a child with cancer could be identified. 
. IMPLICATIONS FOR PRACTICE: Knowledge of the patterns of response will enable nurses to lead parents through a healthy transition process in caring for their children with cancer.

A Cell-Free Assembly System for Generating Infectious Human Papillomavirus 16 Capsids Implicates a Size Discrimination Mechanism for Preferential Viral Genome Packaging.

UNLABELLED: We have established a cell-free in vitro system to study human papillomavirus type 16 (HPV16) assembly, a poorly understood process. L1/L2 capsomers, obtained from the disassembly of virus-like particles (VLPs), were incubated with nuclear extracts to provide access to the range of cellular proteins that would be available during assembly within the host cell. Incorporation of a reporter plasmid "pseudogenome" was dependent on the presence of both nuclear extract and ATP. Unexpectedly, L1/L2 VLPs that were not disassembled prior to incubation with a reassembly mixture containing nuclear extract also encapsidated a reporter plasmid. As with HPV pseudoviruses (PsV) generated intracellularly, infection by cell-free particles assembled in vitro required the presence of L2 and was susceptible to the same biochemical inhibitors, implying the cell-free assembled particles use the infectious pathway previously described for HPV16 produced in cell culture. Using biochemical and electron microscopy analyses, we observed that, in the presence of nuclear extract, intact VLPs partially disassemble, providing a mechanistic explanation to how the exogenous plasmid was packaged by these particles. Further, we provide evidence that capsids containing an <8-kb pseudogenome are resistant to the disassembly/reassembly reaction. Our results suggest a novel size discrimination mechanism for papillomavirus genome packaging in which particles undergo iterative rounds of disassembly/reassembly, seemingly sampling DNA until a suitably sized DNA is encountered, resulting in the formation of a stable virion structure. IMPORTANCE: Little is known about papillomavirus assembly biology due to the difficulties in propagating virus in vitro. The cell-free assembly method established in this paper reveals a new mechanism for viral genome packaging and will provide a tractable system for further dissecting papillomavirus assembly. The knowledge gained will increase our understanding of virus-host interactions, help to identify new targets for antiviral therapy, and allow for the development of new gene delivery systems based on in vitro-generated papillomavirus vectors.

Kallikrein-8 Proteolytically Processes Human Papillomaviruses in the Extracellular Space To Facilitate Entry into Host Cells.

UNLABELLED: The entry of human papillomaviruses into host cells is a complex process. It involves conformational changes at the cell surface, receptor switching, internalization by a novel endocytic mechanism, uncoating in endosomes, trafficking of a subviral complex to the Golgi complex, and nuclear entry during mitosis. Here, we addressed how the stabilizing contacts in the capsid of human papillomavirus 16 (HPV16) may be reversed to allow uncoating of the viral genome. Using biochemical and cell-biological analyses, we determined that the major capsid protein L1 underwent proteolytic cleavage during entry. In addition to a dispensable cathepsin-mediated proteolysis that occurred likely after removal of capsomers from the subviral complex in endosomes, at least two further proteolytic cleavages of L1 were observed, one of which was independent of the low-pH environment of endosomes. This cleavage occurred extracellularly. Further analysis showed that the responsible protease was the secreted trypsin-like serine protease kallikrein-8 (KLK8) involved in epidermal homeostasis and wound healing. Required for infection, the cleavage was facilitated by prior interaction of viral particles with heparan sulfate proteoglycans. KLK8-mediated cleavage was crucial for further conformational changes exposing an important epitope of the minor capsid protein L2. Occurring independently of cyclophilins and of furin that mediate L2 exposure, KLK8-mediated cleavage of L1 likely facilitated access to L2, located in the capsid lumen, and potentially uncoating. Since HPV6 and HPV18 also required KLK8 for entry, we propose that the KLK8-dependent entry step is conserved. IMPORTANCE: Our analysis of the proteolytic processing of incoming HPV16, an etiological agent of cervical cancer, demonstrated that the capsid is cleaved extracellularly by a serine protease active during wound healing and that this cleavage was crucial for infection. The cleavage of L1 is one of at least four structural alterations that prime the virus extracellularly for receptor switching, internalization, and possibly uncoating. This step was also important for HPV6 and HPV18, which may suggest that it is conserved among the papillomaviruses. This study advances the understanding of how HPV16 initially infects cells, strengthens the notion that wounding facilitates infection of epidermal tissue, and may help the development of antiviral measures.

Heparin increases the infectivity of Human Papillomavirus type 16 independent of cell surface proteoglycans and induces L1 epitope exposure.

Human Papillomaviruses (HPVs) are the etiological agents of cervical cancer, and HPV-16 is the most prevalent type. Several HPVs require heparan sulfate proteoglycans (HSPGs) for cell binding. Here, we analyse the phenomenon that preincubation of HPV-16 with increasing concentrations of heparin results in partial restoration rather than more efficient inhibition of infection. While corroborating that the HSPGs are cell-binding receptors for HPV-16, heparin-preincubated virus bound to the extracellular matrix (ECM) via laminin-332. Furthermore, the interaction of virions with heparin, a representative of the highly sulfated S-domains of heparan sulfate (HS) chains of HSPGs, allowed HPV-16 infection in the absence of cell surface HSPGs. Therefore, we concluded that specific glycan moieties but not specific HSPG protein backbones are required for infection. The increased binding of an epitope-specific antibody to the viral capsid after heparin binding suggested that initial conformational changes in the HPV-16 virion occur during infection by interaction with'heparin-like' domains of cellular HSPGs. We propose that HS sequences with specific sulfation patterns are required to facilitate HPV-16 infection.

Principles of polyoma- and papillomavirus uncoating.

Virus particles are vehicles for transmission of the viral genetic information between infected and uninfected cells and organisms. They have evolved to self-assemble, to serve as a protective shell for the viral genome during transfer, and to disassemble when entering a target cell. Disassembly during entry is a complex, multi-step process typically termed uncoating. Uncoating is triggered by multiple host-cell interactions. During cell entry, these interactions occur sequentially in different cellular compartments that the viruses pass through on their way to the site of replication. Here, we highlight the general principles of uncoating for two structurally related virus families, the polyoma- and papillomaviruses. Recent research indicates the use of different compartments and cellular interactions for uncoating despite their structural similarity.

Vanadate but not tungstate prevents the fructose-induced increase in GLUT5 expression and fructose uptake by neonatal rat intestine.

Intermediary signals, precociously enhancing GLUT5 transcription in response to perfusion of its substrate, fructose, in the small intestine of neonatal rats, are not known. Because glucose-6-phosphatase (G6Pase), glucose-6-phosphate translocase (G6PT), and fructose-1,6-bisphosphatase (FBPase) expression increases parallel to or precedes that of GLUT5, we investigated the link between these gluconeogenic genes and GLUT5 by using vanadate or tungstate, potent inhibitors of gluconeogenesis. Small intestinal perfusions of 20-d-old rats were performed with fructose alone, fructose + vanadate or tungstate, glucose alone, and glucose + vanadate or tungstate. As expected, fructose, but not glucose nor glucose + inhibitor perfusion, increased GLUT5 mRNA abundance and fructose transport. Fructose perfusion dramatically increased G6Pase mRNA abundance but had no effect on G6Pase activity. In sharp contrast, fructose perfusion did not increase FBPase gene expression but stimulated FBPase activity. Both vanadate and tungstate significantly inhibited G6Pase activity but did not prevent the fructose-induced increases in G6Pase and G6PT gene expression. Perfusion with fructose + vanadate prevented the fructose-induced increases in fructose transport and GLUT5 mRNA abundance, whereas perfusion with fructose + tungstate did not. Interestingly, vanadate, but not tungstate, inhibited the fructose-induced increase in FBPase activity. Thus, vanadate inhibition of fructose-induced increases in FBPase activity paralleled exactly vanadate inhibition of fructose-induced increases in GLUT5 mRNA abundance and activity. Fructose-induced changes in FBPase activity may regulate changes in GLUT5 expression and activity in the small intestine of neonatal rats. The marked increases in intestinal G6Pase and GLUT5 mRNA abundance may be a parallel response to different factors released during fructose perfusion.

Fructose-induced increases in neonatal rat intestinal fructose transport involve the PI3-kinase/Akt signaling pathway.

Expression of rat glucose transporter-5 (GLUT5) is tightly regulated during development. Expression and activity are low throughout the suckling and weaning stages, but perfusion of the small intestinal lumen with fructose solutions during weaning precociously enhances GLUT5 activity and expression. Little is known, however, about the signal transduction pathways involved in the substrate-induced precocious GLUT5 development. We found that wortmannin and LY-294002, inhibitors of phosphatidylinositol 3-kinase (PI3-kinase) specifically inhibited the increase in fructose uptake rate and brush-border GLUT5 protein abundance but not GLUT5 mRNA abundance. Perfusion of EGF, an activator of PI3-kinase, also resulted in a marked wortmannin-inhibitable increase in fructose uptake. Perfusion of fructose for 4 h increased cytosolic immunostaining of phosphatidylinositol-3,4,5-triphosphate (PIP(3)), the primary product of PI3-kinase, mainly in the mid- to upper-villus regions in which the brush-border membrane also stained strongly with GLUT5. Perfusion of glucose for 4 h had little effect on fructose or glucose uptake and PIP(3) or GLUT5 staining. SH-5, an Akt inhibitor, prevented the increase in fructose uptake and GLUT5 protein induced by fructose solutions, and had no effect on glucose uptake. The PI3-kinase/Akt signaling pathway may be involved in the synthesis and/or recruitment to the brush border of GLUT5 transporters by luminal fructose in the small intestine of weaning rats. Increases in fructose transport during the critical weaning period when rats are shifting to a new diet may be modulated by several signaling pathways whose cross talk during development still needs to be elucidated.

Gill tissue recovery after copper exposure and blood parameter responses in the tropical fish Prochilodus scrofa.

Changes in Prochilodus scrofa gill tissue and in blood responses were investigated after 96-h copper exposure and transference to clean water. Gill damage was characterized by epithelial lifting, cell swelling, pavement, chloride and mucous cell proliferation, and blood vessel anomalies. Restoration of gill structure was slow, with no tissue improvements in the first 2 days in clean water. From the 7th to the 15th day, the recovery of gill tissue began to become evident, with complete recovery occurring on the 45th day in clean water. Hematocrit, red blood cells, and hemoglobin concentration showed a significant increase after copper exposure, remaining high until the 7th day after transference to clean water. Plasma Na(+) and Cl(-) concentration decreased significantly and K(+) increased significantly after copper exposure and, on the 7th day in clean water, plasma ions showed no significant difference from those in control fish. Gill tissue restoration took longer than the recovery of blood parameters, possibly implying extra energy needs, which may be critical, depending on the fish's life cycle.