Modelo calgary de avaliação familiar: mulheres em situação de violência e revogação de medidas protetivas / Calgary family assessment model: women in situations of violence and revocation of protective measures / Modelo calgary de evaluación familiar: mujeres en situaciones de violencia y revocación de medidas de protección

RESUMO Objetivo: compreender as fortalezas e fragilidades das famílias de mulheres em situação de violência que revogaram a medida protetiva de urgência. Método: pesquisa descritiva pautada no Modelo Calgary de Avaliação Familiar, no contexto de uma Delegacia da Mulher em um município do interior de São Paulo, Brasil, realizada entre os meses de setembro e novembro de 2021, com quatro famílias de mulheres que revogaram a medida protetiva. A coleta e análise de dados sobre a estrutura, o desenvolvimento e a funcionalidade familiar foram conduzidos segundo o Modelo Calgary. Resultados: as famílias apresentam similaridades que abrangem aspectos socioeconômicos, baixa escolaridade, uso de álcool, relações familiares conflituosas e transgeracionais que, por suas fragilidades, perpetuam o ciclo de violência. Entretanto, os benefícios sociais, o aporte religioso e de familiares se apresentaram como fortalezas. Conclusão: o Modelo Calgary de Avaliação Familiar proporciona direcionamento à enfermagem para propor planos de cuidados consoantes às complexidades da violência doméstica.

ABSTRACT Objective: To understand the strengths and weaknesses of the families of women in situations of violence who have had their emergency protective measures revoked. Method: descriptive research based on the Calgary Family Assessment Model, in the context of a Women's Police Station in a municipality in the interior of São Paulo, Brazil, carried out between September and November 2021, with four families of women who revoked the protective measure. Family structure, development, and functionality data were collected and analyzed according to the Calgary Model. Results: the families have similarities that include socio-economic aspects, low schooling, alcohol use, conflicting family relationships, and transgenerational relationships that, due to their fragility, perpetuate the cycle of violence. However, the social benefits, religious support, and family members were strengths. Conclusion: The Calgary Family Assessment Model guides nurses in proposing care plans consistent with domestic violence's complexities.

RESUMEN Objetivo: Conocer los puntos fuertes y débiles de las familias de las mujeres en situación de violencia a las que se les ha revocado la medida de protección de urgencia. Método: investigación descriptiva basada en el Modelo de Evaluación Familiar de Calgary, en el contexto de una Comisaría de la Mujer de un municipio del interior de São Paulo, Brasil, realizada entre septiembre y noviembre de 2021, con cuatro familias de mujeres a las que se les revocó la medida de protección. Los datos sobre la estructura, el desarrollo y la funcionalidad de la familia se recopilaron y analizaron utilizando el Modelo de Calgary. Resultados: las familias presentan similitudes que incluyen aspectos socioeconómicos, baja escolarización, consumo de alcohol, relaciones familiares conflictivas y relaciones transgeneracionales que, debido a su fragilidad, perpetúan el ciclo de la violencia. Sin embargo, los beneficios sociales, el apoyo religioso y el apoyo de los miembros de la familia fueron puntos fuertes. Conclusión: El modelo de evaluación familiar de Calgary proporciona una guía para que las enfermeras propongan planes de cuidados acordes con las complejidades de la violencia doméstica.

Contributions of viral oncogenes of HPV-18 and hypoxia to oxidative stress and genetic damage in human keratinocytes.

Infection with high-risk human papillomaviruses like HPV-16 and HPV-18 is highly associated with the development of cervical and other cancers. Malignant transformation requires viral oncoproteins E5, E6 and E7, which promote cell proliferation and increase DNA damage. Oxidative stress and hypoxia are also key factors in cervical malignant transformation. Increased levels of reactive species of oxygen (ROS) and nitrogen (RNS) are found in the hypoxic tumor microenvironment, promoting genetic instability and invasiveness. In this work, we studied the combined effect of E5, E6 and E7 and hypoxia in increasing oxidative stress and promoting DNA damage and nuclear architecture alterations. HaCaT cells containing HPV-18 viral oncogenes (HaCaT E5/E6/E7-18) showed higher ROS levels in normoxia and higher levels of RNS in hypoxia compared to HaCaT parental cells, as well as higher genetic damage in hypoxia as measured by γH2AX and comet assays. In hypoxia, HaCaT E5/E6/E7-18 increased its nuclear dry mass and both cell types displayed marked heterogeneity in nuclear dry mass distribution and increased nuclear foci. Our results show contributions of both viral oncogenes and hypoxia to oxidative stress, DNA damage and altered nuclear architecture, exemplifying how an altered microenvironment combines with oncogenic transformation to promote tumor progression.

Computational multifocus fluorescence microscopy for three-dimensional visualization of multicellular tumor spheroids.

SIGNIFICANCE: Three-dimensional (3D) visualization of multicellular tumor spheroids (MCTS) in fluorescence microscopy can rapidly provide qualitative morphological information about the architecture of these cellular aggregates, which can recapitulate key aspects of their in vivo counterpart. AIM: The present work is aimed at overcoming the shallow depth-of-field (DoF) limitation in fluorescence microscopy while achieving 3D visualization of thick biological samples under study. APPROACH: A custom-built fluorescence microscope with an electrically focus-tunable lens was developed to optically sweep in-depth the structure of MCTS. Acquired multifocus stacks were combined by means of postprocessing algorithms performed in the Fourier domain. RESULTS: Images with relevant characteristics as extended DoF, stereoscopic pairs as well as reconstructed viewpoints of MCTS were obtained without segmentation of the focused regions or estimation of the depth map. The reconstructed images allowed us to observe the 3D morphology of cell aggregates. CONCLUSIONS: Computational multifocus fluorescence microscopy can provide 3D visualization in MCTS. This tool is a promising development in assessing the morphological structure of different cellular aggregates while preserving a robust yet simple optical setup.

Cdk5-Dependent Phosphorylation of CaV3.2 T-Type Channels: Possible Role in Nerve Ligation-Induced Neuropathic Allodynia and the Compound Action Potential in Primary Afferent C Fibers.

Voltage-gated T-type Ca2+ (CaV3) channels regulate diverse physiological events, including neuronal excitability, and have been linked to several pathological conditions such as absence epilepsy, cardiovascular diseases, and neuropathic pain. It is also acknowledged that calcium/calmodulin-dependent protein kinase II and protein kinases A and C regulate the activity of T-type channels. Interestingly, peripheral nerve injury induces tactile allodynia and upregulates CaV3.2 channels and cyclin-dependent kinase 5 (Cdk5) in dorsal root ganglia (DRG) and spinal dorsal horn. Here, we report that recombinant CaV3.2 channels expressed in HEK293 cells are regulatory targets of Cdk5. Site-directed mutagenesis showed that the relevant sites for this regulation are residues S561 and S1987. We also found that Cdk5 may regulate CaV3.2 channel functional expression in rats with mechanical allodynia induced by spinal nerve ligation (SNL). Consequently, the Cdk5 inhibitor olomoucine affected the compound action potential recorded in the spinal nerves, as well as the paw withdrawal threshold. Likewise, Cdk5 expression was upregulated after SNL in the DRG. These findings unveil a novel mechanism for how phosphorylation may regulate CaV3.2 channels and suggest that increased channel activity by Cdk5-mediated phosphorylation after SNL contributes nerve injury-induced tactile allodynia.SIGNIFICANCE STATEMENT Neuropathic pain is a current public health challenge. It can develop as a result of injury or nerve illness. It is acknowledged that the expression of various ion channels can be altered in neuropathic pain, including T-type Ca2+ channels that are expressed in sensory neurons, where they play a role in the regulation of cellular excitability. The present work shows that the exacerbated expression of Cdk5 in a preclinical model of neuropathic pain increases the functional expression of CaV3.2 channels. This finding is relevant for the understanding of the molecular pathophysiology of the disease. Additionally, this work may have a substantial translational impact, since it describes a novel molecular pathway that could represent an interesting therapeutic alternative for neuropathic pain.

Using a variant of coverslip hypoxia to visualize tumor cell alterations at increasing distances from an oxygen source.

Early stages in tumor development involve growth in confined spaces, where oxygen diffusion is limited and metabolic waste products accumulate. This hostile microenvironment imposes strong selective pressures on tumor cells, leading eventually to the survival and expansion of aggressive subclones that condition further tumor evolution. To model features of this microenvironment in vitro, a diffusional barrier can be introduced in the form of a coverslip placed on top of cells, a method termed coverslip hypoxia. Using a variant of this method, with larger volume between coverslip and cells and with oxygen diffusion occurring only through a small hole in the center of the coverslip, we have visualized alterations in LNCaP tumor cells as a function of their distance to the oxygen source at the center. We observed remarkable morphological changes in LNCaP cells as the distance from the center increases, with cells becoming highly spread, displaying dynamic membrane protrusions and occasionally adopting a migratory phenotype. Concomitantly, cells farther from the center displayed marked increases in the hypoxia marker hypoxyprobe, whereas extracellular pH decreased in the same direction. Cells with altered morphology displayed prominent increases in fibrillar actin, as well as swollen mitochondria with distorted cristae and accumulation of neutral lipid-containing intracellular vesicles. These results show that an in vitro microenvironment that models diffusional barriers encountered by tumors in situ can have profound effects on tumor cells. The coverslip hypoxia variant we describe can be used to characterize in vitro the response of tumor cells to environmental conditions that play crucial roles in early tumor development.

Edge enhancement of color images using a digital micromirror device.

A method for orientation-selective enhancement of edges in color images is proposed. The method utilizes the capacity of digital micromirror devices to generate a positive and a negative color replica of the image used as input. When both images are slightly displaced and imagined together, one obtains an image with enhanced edges. The proposed technique does not require a coherent light source or precise alignment. The proposed method could be potentially useful for processing large image sequences in real time. Validation experiments are presented.

Color encoding of binary fringes for gamma correction in 3-D profiling.

Three-dimensional profiling by sinusoidal fringe projection using PSI-algorithms are distorted by the nonlinear response of digital cameras and commercial video projectors. To solve the problem, we present a fringe generation technique that consists of projecting and acquiring a temporal sequence of strictly binary color patterns, whose (adequately weighted) average leads to sinusoidal fringe patterns with the required number of bits, which allows for a reliable three-dimensional profile using a PSI-algorithm. Validation experiments are presented.

Incoherent optical processor for nondirectional edge enhancement of color images.

We present an optical method for nondirectional edge extraction/enhancement in color images. The method is based on the capability of twisted-nematic LCDs to traduce the image information in changes of the state of polarization of light, which allows us to generate simultaneously two replicas of the digital image displayed on the LCD: a true-color ("positive") image and a complementary-color ("negative") one. In our setup the imaging system consists of a lens plus a pupil mask formed with concentric apertures and orthogonal polarizers. This layout allows us to simultaneously image a well-focused positive replica (due to the circular aperture) superimposed to a slightly defocused negative one (due to the annular aperture). It is not difficult to demonstrate that this generates a nondirectional (Laplacian) edge enhancement. Unlike Fourier, our proposal works with incoherent illumination and does not require precise alignment, and thus, it could be a useful tool for edge extraction/enhancement in large images in real-time applications. Validation experiments are presented.

Optical processing of color images with incoherent illumination: orientation-selective edge enhancement using a modified liquid-crystal display.

We present a novel optical method for edge enhancement in color images based on the polarization properties of liquid-crystal displays (LCD). In principle, a LCD generates simultaneously two color-complementary, orthogonally polarized replicas of the digital image used as input. The currently viewed image in standard LCD monitors and cell phone's screens -which we will refer as the "positive image or true-color image"- is the one obtained by placing an analyzer in front of the LCD, in cross configuration to the back polarizer of the display. The orthogonally polarized replica of this image -the "negative image or complementary-color image"- is absorbed by the front polarizer. In order to generate the positive and negative replica with a slight displacement between them, we used a LCD monitor whose analyzer (originally a linear polarizer) was replaced by a calcite crystal acting as beam displacer. When both images are superimposed laterally displaced across the image plane, one obtains an image with enhanced first-order derivatives along a specific direction. The proposed technique works under incoherent illumination and does not require precise alignment, and thus, it could be potentially useful for processing large color images in real-time applications. Validation experiments are presented.

Three-dimensional profiling with binary fringes using phase-shifting interferometry algorithms.

Three-dimensional shape measurements by sinusoidal fringe projection using phase-shifting interferometry algorithms are distorted by the nonlinear response in intensity of commercial video projectors and digital cameras. To solve the problem, we present a method that consists in projecting and acquiring a temporal sequence of strictly binary patterns, whose (adequately weighted) average leads to a sinusoidal fringe pattern with the required number of bits. Since binary patterns consist of "ones" and "zeros"--and no half-tones are involved--the nonlinear response of the projector and the camera will not play a role, and a nearly unit contrast gray-level sinusoidal fringe pattern is obtained. Validation experiments are presented.

Error catastrophe for viruses infecting cells: analysis of the phase transition in terms of error classes.

RNA viruses offer a very exciting arena in which to study evolution in 'real time' owing to both their high replication rate-many generations per day are possible-and their high mutation rate, leading to a large phenotypic variety. They can be regarded as a swarm of genetically related mutants around a dominant or master genetic sequence. This system is called a 'viral quasi-species'. Thus, a common framework to describe RNA viral dynamics is by means of the quasi-species equation (QSE). The QSE is in fact a system of a very large number of nonlinear coupled equations. Here, we consider a simpler formulation in terms of 'error classes', which groups all the sequences differing from the master sequence by the same number of genomic differences into one population class. From this, based on the analogies with Bose condensation, we use thermodynamic inspired observables to analyse and characterize the 'phase transition' through the so-called 'RNA virus error catastrophe'.