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The quantification of human DNA extracts from forensic samples plays a key role in the forensic genetics process, ensuring maximum efficiency and avoiding repeated analyses, over-amplified samples, or unnecessary examinations. In our laboratory, we use the Quantifiler® Trio system to quantify DNA extracts from a wide range of samples extracted from traces (bloodstains, saliva, semen, tissues, etc.), including swabs from touched objects, which are very numerous in the forensic context. This method has been extensively used continuously for nine years, following an initial validation process, and is part of the ISO/IEC 17025 accredited method. In routine practice, based on the quantitative values determined from the extracts of each trace, we use a standard method or a low-copy-number method that involves repeating the amplification with the generation of a consensus genetic profile. Nowadays, when the quantification results are less than 0.003 ng/μL in the minimum extraction volume (40 μL), we do not proceed with the DNA extract analysis. By verifying the limits of the method, we make a conscious cost-benefit choice, in particular by using the least amount of DNA needed to obtain sufficiently robust genetic profiles appropriate for submission to the Italian DNA Forensic Database. In this work, we present a critical re-evaluation of this phase of the method, which is based on the use of standard curves obtained from the average values of the control DNA analysed in duplicate. Considering the various contributions to uncertainty that are difficult to measure, such as manual pipetting or analytical phases carried out by different operators, we have decided to thoroughly investigate the contribution of variability in the preparation of calibration curves to the final results. Thus, 757 samples from 20 independent experiments were re-evaluated using two different standards for the construction of curves, determining the quantitative differences between the two methods. The experiments also determined the parameters of the slope, Y-intercept, R2, and the values of the synthetic control probe to verify how these parameters can provide information on the final outcome of each analysis. The outcome of this revalidation demonstrated that it is preferable to use quantification ranges rather than exact quantitative limits before deciding how to analyse the extracts via PCR or forgoing the determination of profiles. Additionally, we present some preliminary data related to the analysis of samples that would not have been analysed based on the initial validation, from which genetic profiles were obtained after applying a concentration method to the extracts. Our goal is to improve the accredited analytical method, with a careful risk assessment as indicated by accreditation standards, ensuring that no source of evidence is lost in the reconstruction of a criminal event.

Angiotensin-converting enzyme 2 (ACE2) and accessory proteases (TMPRSS2 and CTSL) are needed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cellular entry, and their expression may shed light on viral tropism and impact across the body. We assessed the cell-type-specific expression of ACE2, TMPRSS2 and CTSL across 107 single-cell RNA-sequencing studies from different tissues. ACE2, TMPRSS2 and CTSL are coexpressed in specific subsets of respiratory epithelial cells in the nasal passages, airways and alveoli, and in cells from other organs associated with coronavirus disease 2019 (COVID-19) transmission or pathology. We performed a meta-analysis of 31 lung single-cell RNA-sequencing studies with 1,320,896 cells from 377 nasal, airway and lung parenchyma samples from 228 individuals. This revealed cell-type-specific associations of age, sex and smoking with expression levels of ACE2, TMPRSS2 and CTSL. Expression of entry factors increased with age and in males, including in airway secretory cells and alveolar type 2 cells. Expression programs shared by ACE2+TMPRSS2+ cells in nasal, lung and gut tissues included genes that may mediate viral entry, key immune functions and epithelial-macrophage cross-talk, such as genes involved in the interleukin-6, interleukin-1, tumor necrosis factor and complement pathways. Cell-type-specific expression patterns may contribute to the pathogenesis of COVID-19, and our work highlights putative molecular pathways for therapeutic intervention.

Myeloid cells are the predominant cell type in the tumor microenvironment of human and murine glioblastoma (GBM). By generating a mouse model deficient for all monocyte chemoattractant proteins, here the authors show that blocking monocyte recruitment promotes a compensatory neutrophil influx and that concomitant neutrophil inhibition is required to improve survival in GBM preclinical models.

Monocytes and monocyte-derived macrophages (MDMs) from blood circulation infiltrate glioblastoma (GBM) and promote growth. Here, we show that PDGFS-driven GBM cells induce the expression of the potent proinflammatory cytokine IL- 1[beta] in MDM, which engages IL-1R1 in tumor cells, activates the NF-[kappa]B pathway, and subsequently leads to induction of monocyte chemoattractant proteins (MCPs). Thus, a feedforward paracrine circuit of IL-1[beta]/IL-1R1 between tumors and MDM creates an interdependence driving PDGFS-driven GBM progression. Genetic loss or locally antagonizing IL-1[beta]/IL-1R1 leads to reduced MDM infiltration, diminished tumor growth, and reduced exhausted [CD8.sup.+] T cells and thereby extends the survival of tumor-bearing mice. In contrast to IL-1[beta], IL-1[alpha] exhibits antitumor effects. Genetic deletion of ll1a/b is associated with decreased recruitment of lymphoid cells and loss-of-interferon signaling in various immune populations and subsets of malignant cells and is associated with decreased survival time of PDGFS-driven tumor-bearing mice. In contrast to PDGFB-driven GBM, Nf1-silenced tumors have a constitutively active NF-[kappa]B pathway, which drives the expression of MCPs to recruit monocytes into tumors. These results indicate local antagonism of IL-1[beta] could be considered as an effective therapy specifically for proneural GBM., Introduction Glioblastoma (GBM) is the most prevalent and aggressive primary brain tumor in children and adults and has poor overall survival rates. Numerous therapies have entered clinical trials, with only [...]

ABSTRACT Objective: To understand the perceptions of nurses about the repercussions on the care for children with mental disorders in a pediatric inpatient unit amidst the COVID-19 pandemic. Method: Qualitative research conducted with 13 nurses in a general pediatrics unit in a large hospital in southern Brazil. Data were collected through semi-structured interviews and analyzed according to the Thematic Analysis. Results: Respondents perceived repercussions on hospital routines due to the pandemic, such as restriction of recreation activities; enforcement of social distancing; limitation of family visits; mandatory use of personal protective equipment and difficulty in counter-referral of care. Final considerations: The results characterized the moment experienced in the unit, providing the institution’s managers with tools for the construction and implementation of new practices, making it possible to meet the demands of mental health care for children with mental disorders within the premises of psychosocial care.

Late prenatal development of the human neocortex encompasses a critical period of gliogenesis and cortical expansion. Here, authors use human transcriptomics to capture transience and diversity of cells in middle and late prenatal development, including glial progenitor signatures.

The changes in super-enhancer (SE) landscape of cancers are mainly attributed to cell-intrinsic genomic alterations. Here, the authors perform epigenomic profiling on primary colorectal cancers (CRCs) and their matched normal tissues and show that local tumour microenvironment induces a SE activation and that its target, PDZK1IP1 promotes CRC growth.

ABSTRACT Objective: To reflect on the mental health of nurses who are women and mothers in the context of the COVID-19 pandemic. Method: Theoretical-reflective study based on scientific literature, supported by national and international scientific literature, in addition to a critical analysis by the authors. Results: Reflections on the subject go beyond the impact of motherhood on these women's lives, as it portrays a gender issue and the role of women within society. The stress of working on the frontlines of a pandemic, added to the demands of children and housework, can trigger exhaustion and mental health repercussions. Conclusion: Workers must seek individual measures and health managers must enable collective strategies in the work environment of the institutions, proposing public policies that make both employers, workers and their families co-responsible.

Introduction: Considering the lack of specific treatments to neuropathic pain, this study aimed to evaluate the effect of a single dose adenosine A3 receptor IB-MECA in the inflammatory and neurotrophic parameters of rats submitted to a neuropathic pain model. Methods: 64 adults male Wistar rats were used. Neuropathic pain was induced by the chronic constriction injury (CCI) of sciatic nerve and the treatment consisted in one dose of 0.5 μmol/kg of a selective agonist of adenosine A3 receptor IB-MECA dissolved in 3% DMSO; vehicle groups received DMSO 3% in saline; morphine groups received 5mg/kg Cerebral cortex and hippocampus IL-1β, BDNF and NGF levels were determined by ELISA assay. Results: The key finding was that a single dose of IB-MECA was able to modulate the IL-1β hippocampus levels CCI and the DMSO increased IL-1β and NGF hippocampus levels in sham animals; however, when the DMSO as an IB-MECA vehicle, this effect was not observed, indicating that IB-MECA was able to prevent the effect of DMSO. Conclusions: Considering that the IL-1β role in neuropathic pain is quite explored, as well as the hippocampus contributions, our result corroborates the relationship of A3 receptor and the chronic pain maintenance process. @font-face {font-family:"Cambria Math"; panose-1:2 4 5 3 5 4 6 3 2 4; mso-font-charset:0; mso-generic-font-family:roman; mso-font-pitch:variable; mso-font-signature:-536870145 1107305727 0 0 415 0;}p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-unhide:no; mso-style-qformat:yes; mso-style-parent:""; margin:0in; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Times New Roman",serif; mso-fareast-font-family:"Times New Roman";}.MsoChpDefault {mso-style-type:export-only; mso-default-props:yes;}div.WordSection1 {page:WordSection1;}

Abstract Background By the use of complete or partial dental prosthesis, function, esthetics, and phonetics are reestablished. Few population-based studies are available in the literature using an older adult population. Objective Evaluate the use of and need for dental prostheses and their associated factors. Method This study included a random sample of 287 elderly, in the city of Cruz Alta, Brazil. Use of and need for dental prostheses were the main outcomes, and two analytical models were constructed. Subgroup analyses were performed for individuals using fixed partial denture (FPD) and removable (partial and/or complete) dental prosthesis (RDP). Results Use of and need for dental prosthesis were identified in 83.6% (n=240) and 42.5% (n=122) of the elderly, respectively. Women presented a significantly higher prevalence ratio (PR) for use of dental prostheses (PR=1.15; 95%IC:1.02–1.28) and lower need (PR=0.70; 95%IC:0.54–0.91). No statistically significant difference was demonstrated for the use of FPD regarding the sex. Higher education level was associated with lower use of prostheses (PR=0.73; 95%IC:0.58–0.91) when compared to lower level of education. Medium level of education showed a significantly lower need for dental prostheses (PR=0.61 – 95%IC:0.39–0.94). Those that did not consume alcohol presented with a significantly lower use of FPD and RDP (PR=0.46; 95%IC:0.23–0.93 and 0.90;0.81–0.99, respectively). Conclusion The use of and need for dental prostheses were associated with sex, level of education, and behavioral factors.

Background: Behavioral alterations, like mechanical and thermal hyperalgesia, and modulation of biomarkers in the peripheral and central nervous systems (CNS) are markers of chronic pain. Transcranial direct current stimulation (tDCS) with exercise is a promising therapy for pain due to its neuromodulatory capacity. Objective: To assess the individual effects of tDCS, exercise, and the two combined on the nociceptive response and BDNF, IL-1β, and IL-4 levels in the CNS structures of rats in a chronic pain model. Methods: For 8 consecutive days after the establishment of chronic neuropathic pain by inducing a constriction injury to the sciatic nerve (CCI), the rats received tDCS, exercise, or both treatments combined (20 min/day). The hyperalgesic response was assessed by von Frey and hot plate tests at baseline, 7, and 14 days after CCI surgery and immediately, 24 h, and 7 days after the end of treatment. The BDNF, IL-1β, and IL-4 levels were assessed in the cerebral cortex, brainstem, and spinal cord by enzyme-linked immunosorbent assay at 48 h and 7 days after the end of treatment. Results: The CCI model triggered marked mechanical and thermal hyperalgesia. However, bimodal tDCS, aerobic exercise, and the two combined relieved nociceptive behavior for up to 7 days following treatment completion. Conclusions: Bimodal tDCS, aerobic exercise, or both treatments combined promoted analgesic effects for neuropathic pain. Such effects were reflected by cytokine modulation throughout the spinal cord-brainstem-cerebral cortex axis.

Objective: to analyze the aspects that hinder blood donation and strategies to attract donors. Methods: qualitative study conducted with 12 donors. The data was collected through semi-structured interviews and analyzed using Content Analysis. Results: regarding the complicating aspects, the lack of time and little flexibility in service hours, the displacement in accessing the service and the fear of the donation process were scored. As strategies for attraction, the interviewees talked about the dissemination and internal campaigns in companies, education in schools and universities, loyalty of donors, flexibility in the hours of the blood center and mobile units for blood donation at strategic points in the city. Conclusion: donors pointed out difficulties and suggestions for strategies, constituting relevant information for innovative proposals that help in the process of attracting new blood donors.

Objective: to understand family members’ perceptions of nursing care for people with psychiatric symptoms. Methods: qualitative study, with 13 family members, in five clinical inpatient units linked to the clinical nursing service of a general hospital. Data were collected through semi-structured interviews. Results: the challenges perceived by family members were turnover of professionals in the work schedules and difficulty in specialized management. The participants suggested training of the team, greater multi-professional integration, improved management and reduced turnover of professionals during care. Conclusion: family members verbalized difficulties and suggestions to support reflection on the care offered to people with psychiatric symptoms hospitalized in clinical units, in order to improve work practices and qualify care.

ABSTRACT Objective: To reflect on the mental health of Nursing professionals in the context of the coronavirus pandemic. Method: This is a theoretical-reflective study based on the discursive formulation on the theme and supported by the national and international scientific literature and by the authors' critical analysis. Results: The analyzed studies, along with the care practice, showed that Nursing professionals are susceptible to the exacerbation of symptoms such as depression, anxiety, insomnia, anguish, and stress in the midst of the coronavirus pandemic, in view of their exhaustive work shifts, patients' deaths, risk of infecting themselves and their families, and social isolation. Final considerations: The mental health of Nursing professionals needs to be listed as one of the priorities for health managers, guaranteeing strategies and public policies that ensure sanity for those who are in the front line of the fight against the pandemic.

Monocytes and monocyte-derived macrophages (MDMs) from blood circulation infiltrate glioblastoma (GBM) and promote growth. Here, we show that PDGFB-driven GBM cells induce the expression of the potent proinflammatory cytokine IL-1ß in MDM, which engages IL-1R1 in tumor cells, activates the NF-κB pathway, and subsequently leads to induction of monocyte chemoattractant proteins (MCPs). Thus, a feedforward paracrine circuit of IL-1ß/IL-1R1 between tumors and MDM creates an interdependence driving PDGFB-driven GBM progression. Genetic loss or locally antagonizing IL-1ß/IL-1R1 leads to reduced MDM infiltration, diminished tumor growth, and reduced exhausted CD8+ T cells and thereby extends the survival of tumor-bearing mice. In contrast to IL-1ß, IL-1α exhibits antitumor effects. Genetic deletion of Il1a/b is associated with decreased recruitment of lymphoid cells and loss-of-interferon signaling in various immune populations and subsets of malignant cells and is associated with decreased survival time of PDGFB-driven tumor-bearing mice. In contrast to PDGFB-driven GBM, Nf1-silenced tumors have a constitutively active NF-κB pathway, which drives the expression of MCPs to recruit monocytes into tumors. These results indicate local antagonism of IL-1ß could be considered as an effective therapy specifically for proneural GBM. [ABSTRACT FROM AUTHOR]