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Growth-associated protein of 43 kDa (GAP43) is a key cytoskeleton-associated component of the presynaptic terminal that facilitates neuroplasticity. Downregulation of GAP43 expression has been associated to various psychiatric conditions in humans and evokes hippocampus-dependent memory impairments in mice. Despite the extensive studies conducted on hippocampal GAP43 in past decades, however, very little is known about its roles in modulating the excitatory vs. inhibitory balance in other brain regions. We recently generated conditional knockout mice in which the Gap43 gene was selectively inactivated in either telencephalic glutamatergic neurons (Gap43fl/fl ;Nex1Cre mice, hereafter Glu-GAP43-/- mice) or forebrain GABAergic neurons (Gap43fl/fl ;Dlx5/6Cre mice, hereafter GABA-GAP43-/- mice). Here, we show that Glu-GAP43-/- but not GABA-GAP43-/- mice of either sex show a striking hyperactive phenotype when exposed to a novel environment. This behavioral alteration of Glu-GAP43-/- mice was linked to a selective activation of dorsal-striatum neurons, as well as to an enhanced corticostriatal glutamatergic transmission and an abrogation of corticostriatal endocannabinoid-mediated long-term depression. In line with these observations, GAP43 was abundantly expressed in corticostriatal glutamatergic terminals of wild-type mice. The novelty-induced hyperactive phenotype of Glu-GAP43-/- mice was abrogated by chemogenetically inhibiting corticostriatal afferences with a Gi-coupled "designer receptor exclusively activated by designer drugs" (DREADD), thus further supporting that novelty-induced activity is controlled by GAP43 at corticostriatal excitatory projections. Taken together, these findings show an unprecedented regulatory role of GAP43 in the corticostriatal circuitry and provide a new mouse model with a delimited neuronal-circuit alteration for studying novelty-induced hyperactivity, a phenotypic shortfall that occurs in diverse psychiatric diseases.Significance statement Psychiatric alterations such as attention deficit/hyperactivity disorder, schizophrenia and bipolar disorder pose a significant health and socioeconomic burden to our society. Animal models that recapitulate precise phenotypical traits of those diseases are therefore warranted for developing new therapeutic interventions. Here, we found that mice lacking the protein GAP43 selectively in telencephalic glutamatergic neurons show a robust novelty-induced hyperactive phenotype, a behavioral deficit often associated to psychiatric diseases. These mice exhibit profound alterations in corticostriatal excitatory plasticity and a selective overactivation of dorsal-striatum neurons in response to a novel environment. Our findings thus unveil an important role of GAP43 in corticostriatal function and provide a new animal model with a delimited neuronal-circuit alteration for studying novelty-induced hyperactivity in psychiatric disorders.
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Osteoarthritis (OA) is characterized by an abundance of inflammatory M1-like macrophages damaging local tissues. The search for new potential drugs for OA suffers from the lack of appropriate methods of long-lasting inflammation. Here we developed and characterized an in vitro protocol of long-lasting culture of primary human monocyte-derived macrophages differentiated with a combination of M-CSF+GM-CSF that optimally supported long-cultured macrophages (LC-MÏs) for up to 15 days, unlike their single use. Macrophages repeatedly stimulated for 15 days with the TLR2 ligand Pam3CSK4 (LCS-MÏs), showed sustained levels over time of IL-6, CCL2, and CXCL8, inflammatory mediators that were also detected in the synovial fluids of OA patients. Furthermore, macrophages isolated from the synovia of two OA patients showed an expression profile of inflammation-related genes similar to that of LCS-MÏs, validating our protocol as a model of chronically activated inflammatory macrophages. Next, to confirm that these LCS-MÏs could be modulated by anti-inflammatory compounds, we employed dexamethasone and/or celecoxib, two drugs widely used in OA treatment, that significantly inhibited the production of inflammatory mediators. This easy-to-use in vitro protocol of long-lasting inflammation with primary human macrophages could be useful for the screening of new compounds to improve the therapy of inflammatory disorders.
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Osteoartrite , Agonistas do Receptor Semelhante a Toll , Humanos , Macrófagos/metabolismo , Osteoartrite/tratamento farmacológico , Osteoartrite/metabolismo , Inflamação/metabolismo , Mediadores da Inflamação/metabolismoRESUMO
Cu+ -chaperones are a diverse group of proteins that allocate Cu+ ions to specific copper proteins, creating different copper pools targeted to specific physiological processes. Symbiotic nitrogen fixation carried out in legume root nodules indirectly requires relatively large amounts of copper, for example for energy delivery via respiration, for which targeted copper deliver systems would be required. MtNCC1 is a nodule-specific Cu+ -chaperone encoded in the Medicago truncatula genome, with a N-terminus Atx1-like domain that can bind Cu+ with picomolar affinities. MtNCC1 is able to interact with nodule-specific Cu+ -importer MtCOPT1. MtNCC1 is expressed primarily from the late infection zone to the early fixation zone and is located in the cytosol, associated with plasma and symbiosome membranes, and within nuclei. Consistent with its key role in nitrogen fixation, ncc1 mutants have a severe reduction in nitrogenase activity and a 50% reduction in copper-dependent cytochrome c oxidase activity. A subset of the copper proteome is also affected in the ncc1 mutant nodules. Many of these proteins can be pulled down when using a Cu+ -loaded N-terminal MtNCC1 moiety as a bait, indicating a role in nodule copper homeostasis and in copper-dependent physiological processes. Overall, these data suggest a pleiotropic role of MtNCC1 in copper delivery for symbiotic nitrogen fixation.
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Medicago truncatula , Fixação de Nitrogênio , Fixação de Nitrogênio/genética , Medicago truncatula/genética , Medicago truncatula/metabolismo , Cobre/metabolismo , Nódulos Radiculares de Plantas/metabolismo , Simbiose/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Chronic inflammatory diseases are increasing in developed societies, thus new anti-inflammatory approaches are needed in the clinic. Synthetic peptides complexes can be designed to mimic the activity of anti-inflammatory mediators, in order to alleviate inflammation. Here, we evaluated the anti-inflammatory efficacy of tethered peptides mimicking the interleukin-1 receptor antagonist (IL-1Ra) and the heat-shock protein 70 (HSP70). We tested their biocompatibility and anti-inflammatory activity in vitro in primary human monocytes and differentiated macrophages activated with two different stimuli: the TLR agonists (LPS + IFN-γ) or Pam3CSK4. Our results demonstrate that IL-1Ra and HSP70 synthetic peptides present a satisfactory biocompatible profile and significantly inhibit the secretion of several pro-inflammatory cytokines (IL-6, IL-8, IL-1ß and TNFα). We further confirmed their anti-inflammatory activity when peptides were coated on a biocompatible material commonly employed in surgical implants. Overall, our findings support the potential use of IL-1Ra and HSP70 synthetic peptides for the treatment of inflammatory conditions.
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Anti-Inflamatórios , Proteína Antagonista do Receptor de Interleucina 1 , Humanos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Proteína Antagonista do Receptor de Interleucina 1/farmacologia , Proteína Antagonista do Receptor de Interleucina 1/uso terapêutico , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Peptídeos/farmacologia , Peptídeos/uso terapêuticoRESUMO
OBJECTIVE: The purpose of this study was to assess the visual performance and monochromatic higher-order aberrations (HOAs) obtained while wearing a MiSight dual-focus (DF) contact lenses (CL) in comparison with a single-vision contact lens (SVCL). METHODS: A randomized, double-masked, cross-over study was performed. Participants were fitted with a DFCL and a SVCL composed of the same material (omafilcon A) and parameters. Logarithm of the Minimum Angle of Resolution high-contrast (100%) and low-contrast (10%) visual acuity (VA) and contrast sensitivity (CS) for 3, 6, 12, and 18 cycles per degree were measured. Higher-order aberrations were also evaluated using a Hartmann-Shack aberrometer with the CLs on. RESULTS: Twenty-four subjects (21 females and 3 males) with a mean age of 21.9±1.9 years (range: 18-27) were included. Low-contrast VA was significantly lower with the DFCL regarding the SVCL design (0.39±0.23 vs 0.25±0.18, P=0.002). However, there were no differences in high-contrast VA between both CLs (-0.03±0.10 vs -0.09±0.14, P=0.187). Contrast sensitivity was lower with the DFCL under all spatial frequencies (P≤0.048). Second-, third-, fourth-, and fifth-order aberrations were significantly (P<0.001) higher for the DFCL. There were also significant differences between DFCL and SVCL in defocus (0.87±0.28 vs 0.16±0.35, P<0.001), oblique trefoil (-0.16±0.27 vs -0.01±0.08, P=0.005), vertical coma (0.13±0.17 vs 0.00±0.08, P=0.002), and spherical aberration (0.09±0.11 vs -0.02±0.05, P=0.002). CONCLUSION: Visual performance for detecting low-contrast targets is reduced when wearing MiSight DFCL compared with a SVCL with the same material. The main reason might be the induction of second-order and HOAs by the DFCL design.
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Lentes de Contato Hidrofílicas , Lentes de Contato , Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Sensibilidades de Contraste , Estudos Cross-Over , Transtornos da Visão , Acuidade Visual , AdolescenteRESUMO
Structural Health Monitoring (SHM) systems allow three types of diagnostic tasks to be performed, namely damage identification, loads monitoring, and damage prognosis. Only if all three tasks are correctly fulfilled can the useful remaining life of a structure be estimated credibly. This paper deals with the second task and aimed to extend state-of-the-art in load identification, by demonstrating that it is feasible to achieve it through the analysis of response signals captured with high-speed three-dimensional Digital Image Correlation (HS 3D-DIC). The efficacy of the proposed procedure is demonstrated experimentally on a frame structure under broadband vibration excitation. Full-field vibration displacement signals are captured with the use of two high-speed cameras and processed with 3D-DIC. Loads are identified with two different algorithms based on inverting the Frequency Response Function (FRF) matrix and modal filtration (MF). The paper discusses both methods providing their theoretical background and experimental performance.
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Algoritmos , Imageamento Tridimensional , Imageamento Tridimensional/métodos , Vibração , EletrocardiografiaRESUMO
Infections caused by ceftolozane-tazobactam and ceftazidime-avibactam-resistant P. aeruginosa infections are an emerging concern. We aimed to analyze the underlying ceftolozane-tazobactam and ceftazidime-avibactam resistance mechanisms in all multidrug-resistant or extensively drug-resistant (MDR/XDR) P. aeruginosa isolates recovered during 1 year (2020) from patients with a documented P. aeruginosa infection. Fifteen isolates showing ceftolozane-tazobactam and ceftazidime-avibactam resistance were evaluated. Clinical conditions, previous positive cultures, and ß-lactams received in the previous month were reviewed for each patient. MICs were determined by broth microdilution. Multilocus sequence types (MLSTs) and resistance mechanisms were determined using short- and long-read whole-genome sequencing (WGS). The impact of Pseudomonas-derived cephalosporinases (PDCs) on ß-lactam resistance was demonstrated by cloning into an ampC-deficient PAO1 derivative (PAOΔC) and construction of 3D models. Genetic support of acquired ß-lactamases was determined in silico from high-quality hybrid assemblies. In most cases, the isolates were recovered after treatment with ceftolozane-tazobactam or ceftazidime-avibactam. Seven isolates from different sequence types (STs) owed their ß-lactam resistance to chromosomal mutations and all displayed specific substitutions in PDC: Phe121Leu and Gly222Ser, Pro154Leu, Ala201Thr, Gly214Arg, ΔGly203-Glu219, and Glu219Lys. In the other eight isolates, the ST175 clone was overrepresented (6 isolates) and associated with IMP-28 and IMP-13, whereas two ST1284 isolates produced VIM-2. The cloned PDCs conferred enhanced cephalosporin resistance. The 3D PDC models revealed rearrangements affecting residues involved in cephalosporin hydrolysis. Carbapenemases were chromosomal (VIM-2) or plasmid-borne (IMP-28, IMP-13) and associated with class-1 integrons located in Tn402-like transposition modules. Our findings highlighted that cephalosporin/ß-lactamase inhibitors are potential selectors of MDR/XDR P. aeruginosa strains producing PDC variants or metallo-ß-lactamases. Judicious use of these agents is encouraged.
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Ceftazidima , Infecções por Pseudomonas , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Compostos Azabicíclicos/farmacologia , Compostos Azabicíclicos/uso terapêutico , Proteínas de Bactérias , Ceftazidima/farmacologia , Ceftazidima/uso terapêutico , Cefalosporinas/farmacologia , Cefalosporinas/uso terapêutico , Combinação de Medicamentos , Humanos , Testes de Sensibilidade Microbiana , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa , Tazobactam/farmacologia , Tazobactam/uso terapêutico , beta-Lactamases/genética , beta-Lactamases/uso terapêuticoRESUMO
Introduction and Objective: Zygomatic implants (ZI) offer a good and predictable alternative to reconstructive procedures of atrophic maxillae. The main objetive of this systematic review was to assess the effect of rehabilitation with zygomatic implants on patient's quality of life (QLP) using Patient Reported Outcomes Measures (PROMs).Materials and Methods: This review followed PRISMA guidelines. An automated electronic search was conducted in four databases supplemented by a manual search for relevant articles published until the end of January 2021. The Cochrane Collaboration Risk of Bias tool and the Newcastle-Ottawa Quality Assessment Scale were used to assess the quality of evidence in the studies reviewed.Results: General findings of this systematic review showed substantial increases in Oral health-related quality of life (OHRQoL) among patients restored with ZI and high scores in terms of general satisfaction, especially in chewing ability and esthetics. An overall survival rate of ZI was 98.3% after a mean follow-up time of 46.5 months was observed. Occurrence of 13.1% biological complications and 1.8% technical complications were reported.Conclusions: Patients rehabilitated with zygomatic implant-supported complete dental prostheses showed substantial improvements in OHRQoL and general satisfaction with the treatment received.
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Implantes Dentários , Arcada Edêntula , Atrofia/patologia , Implantação Dentária Endóssea/métodos , Prótese Dentária Fixada por Implante , Seguimentos , Humanos , Arcada Edêntula/patologia , Arcada Edêntula/reabilitação , Arcada Edêntula/cirurgia , Maxila/patologia , Maxila/cirurgia , Medidas de Resultados Relatados pelo Paciente , Qualidade de Vida , Resultado do Tratamento , Zigoma/cirurgiaRESUMO
The absence of standardized molecular profiling to differentiate uterine leiomyosarcomas versus leiomyomas represents a current diagnostic challenge. In this study, we aimed to search for a differential molecular signature for these myometrial tumors based on artificial intelligence. For this purpose, differential exome and transcriptome-wide research was performed on histologically confirmed leiomyomas (n = 52) and leiomyosarcomas (n = 44) to elucidate differences between and within these two entities. We identified a significantly higher tumor mutation burden in leiomyosarcomas vs. leiomyomas in terms of somatic single-nucleotide variants (171,863 vs. 81,152), indels (9491 vs. 4098), and copy number variants (8390 vs. 5376). Further, we discovered alterations in specific copy number variant regions that affect the expression of some tumor suppressor genes. A transcriptomic analysis revealed 489 differentially expressed genes between these two conditions, as well as structural rearrangements targeting ATRX and RAD51B. These results allowed us to develop a machine learning approach based on 19 differentially expressed genes that differentiate both tumor types with high sensitivity and specificity. Our findings provide a novel molecular signature for the diagnosis of leiomyoma and leiomyosarcoma, which could be helpful to complement the current morphological and immunohistochemical diagnosis and may lay the foundation for the future evaluation of malignancy risk.
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Leiomioma , Leiomiossarcoma , Neoplasias Uterinas , Inteligência Artificial , Diagnóstico Diferencial , Feminino , Humanos , Leiomioma/diagnóstico , Leiomioma/genética , Leiomioma/metabolismo , Leiomiossarcoma/diagnóstico , Leiomiossarcoma/genética , Leiomiossarcoma/metabolismo , Transcriptoma , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/genética , Neoplasias Uterinas/metabolismoRESUMO
Implementation of the Global Sulphur Cap (GSC), in January 2020, boosted scrubber installation in vessels to fulfill the new air emission limitations. This increase in scrubbers' use has intensified concern about its environmental performance. Even though achievement of GSC requirements through this mitigation system has been widely proven, the impact of wash water discharge on the marine environment remains under discussion. In this paper, an assessment environmental model is introduced to quantify in monetary terms the performance of feeder vessels that operate with several mitigation systems. This model attempts to improve traditional air emission evaluations by including the impact of scrubbers' discharges on the marine environmental. In this way, the analysis not only allows different mitigations systems to be ranked by considering their capacity to reduce air emissions, but also provides further information about the marine eutrophication and ecotoxicity impact from scrubbers' discharge. Through the model's application to a regular shipping line between the Canary Islands and the Iberian Peninsula, it was found that, the scrubber, regardless of its operation mode (open- or closed loop), is the most efficient mitigation option after the Liquefied Natural Gas (LNG) fuel shift. The impact of scrubbers' discharge was not as significant as expected on the feeder vessel's total pollution since this provides similar relative weight to the methane emissions from a dual-engine option by operating with LNG. The results also show the need to more closely research the marine eutrophication impact of closed-loop scrubbers. Finally, this paper warns about a significant dispersion on the monetary values of marine ecotoxicity and eutrophication, due to a high dependence of the results on the frameworks' localization. Consequently, further research is needed on the homogenization of pollution monetization in the marine environment.
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Poluição do Ar , Navios , Poluição Ambiental , Metano/análise , Modelos Teóricos , Gás NaturalRESUMO
The aim of this study was to compare tumour burden in patients who underwent surgery for melanoma and cutaneous squamous cell carcinoma during nationwide lockdown in Spain due to COVID-19 (for the period 14 March to 13 June 2020) and during the same dates in 2019 before the COVID-19 pandemic. In addition, associations between median tumour burden (Breslow thickness for melanoma and maximum clinical diameter for cutaneous squamous cell carcinoma) and demographic, clinical, and medical factors were analysed, building a multivariate linear regression model. During the 3 months of lockdown, there was a significant decrease in skin tumours operated on (41% decrease for melanoma (n = 352 vs n = 207) and 44% decrease for cutaneous squamous cell carcinoma (n = 770 vs n = 429)) compared with the previous year. The proportion of large skin tumours operated on increased. Fear of SARS-CoV-2 infection, with respect to family member/close contact, and detection of the lesion by the patient or doctor, were related to thicker melanomas; and fear of being diagnosed with cancer, and detection of the lesion by the patient or relatives, were related to larger size cutaneous squamous cell carcinoma. In conclusion, lockdown due to COVID-19 has resulted in a reduction in treatment of skin cancer.
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COVID-19 , Carcinoma de Células Escamosas , Melanoma , Neoplasias Cutâneas , Carcinoma de Células Escamosas/epidemiologia , Carcinoma de Células Escamosas/cirurgia , Controle de Doenças Transmissíveis , Humanos , Melanoma/epidemiologia , Melanoma/cirurgia , Pandemias , SARS-CoV-2 , Neoplasias Cutâneas/epidemiologia , Neoplasias Cutâneas/cirurgia , Carga TumoralRESUMO
Experimental characterization and validation of skin components in aircraft entails multiple evaluations (structural, aerodynamic, acoustic, etc.) and expensive campaigns. They require different rigs and equipment to perform the necessary tests. Two of the main dynamic characterizations include the energy absorption under impact forcing and the identification of modal parameters through the vibration response under any broadband excitation, which also includes impacts. This work exploits the response of a stiffened aircraft composite panel submitted to a multi-impact excitation, which is intended for impact and energy absorption analysis. Based on the high stiffness of composite materials, the study worked under the assumption that the global response to the multi-impact excitation is linear with small strains, neglecting the nonlinear behavior produced by local damage generation. Then, modal identification could be performed. The vibration after the impact was measured by high-speed 3D digital image correlation and employed for full-field operational modal analysis. Multiple modes were characterized in a wide spectrum, exploiting the advantages of the full-field noninvasive techniques. These results described a consistent modal behavior of the panel along with good indicators of mode separation given by the auto modal assurance criterion (Auto-MAC). Hence, it illustrates the possibility of performing these dynamic characterizations in a single test, offering additional information while reducing time and investment during the validation of these structures.
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Uterine leiomyomas represent the most common benign gynecologic tumor. These hormone-dependent smooth-muscle formations occur with an estimated prevalence of ~70% among women of reproductive age and cause symptoms including pain, abnormal uterine bleeding, infertility, and recurrent abortion. Despite the prevalence and public health impact of uterine leiomyomas, available treatments remain limited. Among the potential causes of leiomyomas, early hormonal exposure during periods of development may result in developmental reprogramming via epigenetic changes that persist in adulthood, leading to disease onset or progression. Recent developments in unbiased high-throughput sequencing technology enable powerful approaches to detect driver mutations, yielding new insights into the genomic instability of leiomyomas. Current data also suggest that each leiomyoma originates from the clonal expansion of a single transformed somatic stem cell of the myometrium. In this review, we propose an integrated cellular and molecular view of the origins of leiomyomas, as well as paradigm-shifting studies that will lead to better understanding and the future development of non-surgical treatments for these highly frequent tumors.
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Leiomioma/patologia , Neoplasias Uterinas/patologia , Útero/patologia , Animais , Epigênese Genética , Feminino , Regulação Neoplásica da Expressão Gênica , Predisposição Genética para Doença , Humanos , Leiomioma/etiologia , Leiomioma/genética , Mutação , Neoplasias Uterinas/etiologia , Neoplasias Uterinas/genética , Útero/metabolismoRESUMO
BACKGROUND: The treatment of multiple myeloma (MM) has become costly and difficult to access for patients living in low-income to middle-income countries. METHODS: The current retrospective study included 148 patients in Mexico with newly diagnosed MM, and was performed to compare the outcomes of patients with and without access to novel agents. The records of 77 patients admitted to a public hospital (PubC) and 71 patients cared for within private health systems (PrivC) from November 2007 to July 2016 were reviewed. RESULTS: Compared with those treated in PrivC, patients receiving care at PubC were more likely to be diagnosed with advanced disease. A thalidomide-based regimen was the most common induction treatment used at PubC, whereas a bortezomib-based regimen was used most often in PrivC. The median follow-up was 41 months. Patients in PrivC demonstrated better response rates and survival; 65% of patients treated in PrivC versus 41% treated at PubC achieved a very good partial response or better (P = .005). The median progression-free survival and median overall survival were 23 months and 51 months, respectively, for patients treated at PubC and 41 months and 79 months, respectively, for those treated in PrivC (P<.001). More patients underwent autologous stem cell transplantation in PrivC. When adjustments were made for covariates, patients treated at PubC experienced a higher risk of death compared with patients receiving care in PrivC (hazard ratio, 2.0; 95% confidence interval, 1.0-4.3 [P = .04]). CONCLUSIONS: Stage at diagnosis, induction regimen, and autologous stem cell transplantation were found to be contributors to survival disparities between patients with MM treated at PubC compared with PrivC in Mexico. These findings underscore the need to improve access to novel agents and stem cell transplantation in public health systems. Cancer 2018;124:1946-53. © 2018 American Cancer Society.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Custos de Medicamentos , Acessibilidade aos Serviços de Saúde/economia , Disparidades em Assistência à Saúde/economia , Transplante de Células-Tronco Hematopoéticas/economia , Mieloma Múltiplo/terapia , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/economia , Bortezomib/economia , Bortezomib/uso terapêutico , Intervalo Livre de Doença , Feminino , Seguimentos , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Disparidades em Assistência à Saúde/estatística & dados numéricos , Transplante de Células-Tronco Hematopoéticas/estatística & dados numéricos , Hospitais Privados/economia , Hospitais Privados/estatística & dados numéricos , Hospitais Públicos/economia , Hospitais Públicos/estatística & dados numéricos , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Mieloma Múltiplo/economia , Mieloma Múltiplo/mortalidade , Mieloma Múltiplo/patologia , Estudos Retrospectivos , Talidomida/economia , Talidomida/uso terapêutico , Transplante Autólogo/economia , Transplante Autólogo/estatística & dados numéricos , Resultado do TratamentoRESUMO
Three-dimensional digital image correlation (3D-DIC) has become the most popular full-field optical technique for measuring 3D shapes and displacements in experimental mechanics. The integration of fringe projection (FP) and two-dimensional digital image correlation (FP + DIC) has been recently established as an intelligent low-cost alternative to 3D-DIC, overcoming the drawbacks of a stereoscopic system. Its experimentation is based on the colour encoding of the characterized fringe and speckle patterns required for FP and DIC implementation, respectively. In the present work, innovations in experimentation using FP + DIC for more accurate results are presented. Specifically, they are based on the improvement of the colour pattern encoding. To achieve this, in this work, a multisensor camera and/or laser structural illumination were employed. Both alternatives are analysed and evaluated. Results show that improvements both in three-dimensional and in-plane displacement are obtained with the proposed alternatives. Nonetheless, multisensor high-speed cameras are uncommon, and laser structural illumination is established as an important improvement when low uncertainty is required for 2D-displacement measurement. Hence, the uncertainty has been demonstrated to be reduced by up to 50% compared with results obtained in previous experimental approaches of FP + DIC.
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BACKGROUND: Cisplatin is a potent antitumor agent. However, toxicity and primary and secondary resistance are major limitations of cisplatin-based chemotherapy, leading to therapeutic failure. We have previously reported that mono-sulfonamide platinum complexes have good antitumor activity against different tumoral cell lines and with a different and better cytotoxic profile than cisplatin. Besides, N-sulfonamides have been used extensively in medicinal chemistry as bactericides, anticonvulsant, inhibitors of the carbonic anhydrase, inhibitors of histone deacetylases, and inhibitors of microtubule polymerization, among others. METHODS: We aimed to compare the cytotoxic effects of cisplatin and a trans-sulfonamide-platinum-complex (TSPC), in two human melanoma cell lines that differ in their TP53 status: SK-MEL-5, TP53 wild type, and SK-MEL-28, TP53 mutated. We performed cytotoxicity assays with both drugs, alone and in combination, cell cycle analyses, western blotting and immunoprecipitation, and fluorescence immunocytochemistry. RESULTS: TSPC had similar antiproliferative activity than cisplatin against SK-MEL-5 (3.24 ± 1.08 vs 2.89 ± 1.12 µM) and higher against SK-MEL-28 cells (5.83 ± 1.06 vs 10.17 ± 1.29 µM). Combination of both drugs inhibited proliferation in both cell lines, being especially important in SK-MEL-28, and showing a synergistic effect. In contrast to cisplatin, TSPC caused G1 instead G2/M arrest in both cell lines. Our present findings indicate that the G1 arrest is associated with the induction of CDKN1A and CDKN1B proteins, and that this response is also present in melanoma cells containing TP53 mutated. Also, strong accumulation of CDKN1A and CDKN1B in cells nuclei was seen upon TSPC treatment in both cell lines. CONCLUSIONS: Overall, these findings provide a new promising TSPC compound with in vitro antitumor activity against melanoma cell lines, and with a different mechanism of action from that of cisplatin. Besides, TSPC synergism with cisplatin facilitates its potential use for co-treatment to reduce toxicity and resistance against cisplatin. TSPC remains a promising lead compound for the generation of novel antineoplastic agent and to explore its synergism with other DNA damaging agents.
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Antineoplásicos/farmacologia , Cisplatino/farmacologia , Melanoma/genética , Sulfonamidas/farmacologia , Proteína Supressora de Tumor p53/genética , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Sinergismo Farmacológico , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Melanoma/tratamento farmacológico , Melanoma/metabolismo , Mutação , Compostos Organoplatínicos/farmacologiaRESUMO
To further our understanding about antigen involvement in mantle cell lymphoma (MCL), we analyzed the expression levels of activation-induced cytidine deaminase (AID), a key player in B-cell responses to antigen triggering, in 133 MCL cases; assessed the functionality of AID by evaluating in vivo class switch recombination in 52 MCL cases; and sought for indications of ongoing antigen interactions by exploring intraclonal diversification within 14 MCL cases. The AID full-length transcript and the most frequent splice variants (AID-ΔE4a, AID-ΔE) were detected in 128 (96.2%), 96 (72.2%), and 130 cases (97.7%), respectively. Higher AID full-length transcript levels were significantly associated (P < 0.001) with lack of somatic hypermutation within the clonotypic immunoglobulin heavy variable (IGHV) genes. Median AID transcript levels were higher in lymph node material compared to cases in which peripheral blood was analyzed, implying that clonal behavior is influenced by the microenvironment. Switched tumor-derived IGHV-IGHD-IGHJ transcripts were identified in 5 of 52 cases (9.6%), all of which displayed somatic hypermutation and AID-mRNA expression. Finally, although most cases exhibited low levels of intraclonal diversification, analysis of the mutational activity revealed a precise targeting of somatic hypermutation indicative of an active, ongoing interaction with antigen(s). Collectively, these findings strongly allude to antigen involvement in the natural history of MCL, further challenging the notion of antigen naivety.
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Linfócitos B/metabolismo , Citidina Desaminase/metabolismo , Região Variável de Imunoglobulina , Linfoma de Célula do Manto/metabolismo , Hipermutação Somática de Imunoglobulina , Linfócitos B/imunologia , Linfócitos B/patologia , Humanos , Linfoma de Célula do Manto/imunologia , Linfoma de Célula do Manto/patologiaRESUMO
The importance of protocols for preoperative antisepsis of the hands is given by the risk of transferring bacteria from the hands of the surgical team to the patient during surgery and it is relationship with infection of surgical wound site (SSI). Careful surgical scrub reduces the number of bacteria on the skin, but does not eliminate them completely, remaining transient microorganisms on hands after the surgical scrub. There fore if micropuncture in surgical gloves occurs, the correct preoperative preparation of hands and double gloving will be essential to reduce the risk of bacterial transmission to patients. The protocols for surgical hand antisepsis are two: Surgical scrub with antiseptic soap (hand scrubbing). Surgical scrub by rubbing alcohol (handrubbing). The hand antisepsis by rubbing with an alcohol solution has proved to be significantly more effective compared to soap solutions. We must also see that in surgical hand antisepsis with soap, you must rinse them with water. And often hospitals' taps and keys are contaminated by Pseudomonas spp., including P. aeuinosa.
Assuntos
Desinfecção das Mãos/métodos , Desinfecção das Mãos/normas , Procedimentos Cirúrgicos Operatórios , HumanosRESUMO
Dual-specificity phosphatase 6 (DUSP6/MKP-3) is a mitogen-activated protein kinase phosphatase that regulates extracellular signal-regulated kinases (ERKs) activity via feedback mechanisms, with an increasingly recognized role in tumour biology. The aim of this study was to explore the role of DUSP6 expression in the prognosis of human non-small cell lung cancer (NSCLC). DUSP6 expression levels were evaluated by real-time quantitative reverse transcription polymerase chain reaction (PCR) in 60 NSCLC samples from patients who underwent pulmonary resection at 12 de Octubre University Hospital. We performed a statistical analysis to investigate the correlation of DUSP6 expression and the clinical outcomes. We found that 66.7% of the tumour samples show the downregulation of DUSP6 at the messenger RNA (mRNA) levels compared to benign epithelial lung tissues and 55% of them show at least twofold downregulation of DUSP6 gene expression. Patients were classified into three groups according to their DUSP6 expression levels and those with very low levels (at least twofold downregulation) had the worst outcomes. Using the value of twice below the mean value in benign epithelial lung tissue as a cutoff, the overall survival of patients with very low DUSP6 levels was significantly lower than that in the rest of patients (31.9 ± 18.8 months vs. not reached, P = 0.049). This was most pronounced in adenocarcinoma histology and high-stage tumour samples. Our results suggest that DUSP6 gene expression in tumour samples may be a prognostic marker in NSCLC.