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Mucosal chemokines have antimicrobial properties and play an important role in mucosal immunity. However, little is known about their expression on the ocular surface. This study aimed to analyze the expression of the mucosal chemokines CCL28, CXCL14 and CXCL17 in corneal and conjunctival epithelial cells under in vitro dry eye (DE) conditions, and in conjunctival samples from healthy subjects and DE patients. Human corneal epithelial cells (HCE) and immortalized human conjunctival epithelial cells (IM-HConEpiC) were incubated under hyperosmolar (400-500 mOsM) or inflammatory (TNF-α 25 ng/mL) conditions for 6 h and 24 h to measure CCL28, CXCL14, and CXCL17 gene expression by RT-PCR and their secretion by immunobead-based analysis (CCL28, CXCL14) and ELISA (CXCL17). Additionally, twenty-seven DE patients and 13 healthy subjects were included in this study. DE-related questionnaires (OSDI, mSIDEQ and NRS) evaluated symptomatology. Ocular surface integrity was assessed using vital staining. Tactile sensitivity was measured with Cochet-Bonnet esthesiometer, and mechanic and thermal (heat and cold) sensitivity using Belmonte's non-contact esthesiometer. Subbasal nerve plexus and dendritic cell density were analyzed by in vivo confocal microscopy. Conjunctival cells from participants were collected by impression cytology to measure mucosal chemokines gene expression by RT-PCR. Our results showed that HCE and IM-HConEpiC cells increased CCL28, CXCL14, and CXCL17 secretion under hyperosmolar conditions. The gene expression of CCL28 was significantly upregulated in conjunctival samples from DE patients. CCL28 expression correlated positively with symptomatology, corneal staining, heat sensitivity threshold, and dendritic cell density. CXCL14 expression correlated positively with age, ocular pain, conjunctival staining, tactile sensitivity, and image reflectivity. CXCL17 expression correlated positively with corneal staining. These results suggest that corneal and conjunctival epithelial cells could be a source of CCL28, CXCL14, and CXCL17 on the ocular surface and that CCL28 might be involved in DE pathogenesis.
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Dieldrin/análogos & derivados , Síndromes do Olho Seco , Humanos , Síndromes do Olho Seco/patologia , Quimiocinas/genética , Córnea/patologia , Túnica Conjuntiva/patologia , Quimiocinas CC , Quimiocinas CXCRESUMO
BACKGROUND: Rectal cancer (RC) is a surgical challenge due to its technical complexity. The double-stapled (DS) technique, a standard for colorectal anastomosis, has been associated with notable drawbacks, including a high incidence of anastomotic leak (AL). Low anterior resection with transanal transection and single-stapled (TTSS) anastomosis has emerged to mitigate those drawbacks. METHODS: Observational study in which it described the technical aspects and results of the initial group of patients with medium-low RC undergoing elective laparoscopic total mesorectal excision (TME) and TTSS. RESULTS: Twenty-two patients were included in the series. Favourable postoperative outcomes with a median length of stay of 5 days and an AL incidence of 9.1%. Importantly, all patients achieved complete mesorectal excision with tumour-free margins, and no mortalities were reported. CONCLUSION: TTSS emerges as a promising alternative for patients with middle and lower rectal tumours, offering potential benefits in terms of morbidity reduction and oncological integrity compared with other techniques.
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Canal Anal , Anastomose Cirúrgica , Neoplasias Retais , Grampeamento Cirúrgico , Humanos , Masculino , Feminino , Anastomose Cirúrgica/métodos , Pessoa de Meia-Idade , Idoso , Neoplasias Retais/cirurgia , Canal Anal/cirurgia , Grampeamento Cirúrgico/métodos , Resultado do Tratamento , Reto/cirurgia , Laparoscopia/métodos , Fístula Anastomótica/etiologia , Fístula Anastomótica/prevenção & controle , Adulto , Idoso de 80 Anos ou maisRESUMO
OBJECTIVE: Despite depression being a common comorbidity among adults with cancer, limited literature is available regarding pharmacologic depression treatment patterns and predictors in this population. This study aims to examine patterns and predictors of antidepressant prescribing among adults with cancer and depression in ambulatory care settings in the United States (US). METHODS: This retrospective, cross-sectional study utilized data collected from the 2014 to 2015 National Ambulatory Medical Care Survey (NAMCS). The study sample consisted of adults (age ≥ 18 years) with cancer and depression (unweighted N = 539; weighted N = 11,361,000). A multivariable logistic regression analysis was used to adjust for individual-level factors to identify predictors of antidepressant prescribing. RESULTS: Most patients were adults aged ≥ 65 years, female, and non-Hispanic whites. Thirty-seven percent of the study sample received antidepressant treatment. Multivariable logistic regression analysis revealed that race/ethnicity, physician specialty, and number of medications were significantly associated with receiving antidepressant(s). For example, non-Hispanic whites were two-and-half times more likely to receive an antidepressant [OR 2.43, 95% confidence interval 1.13-5.23] compared to other race/ethnic groups. Every unit increase in the number of prescribed medications increased the likelihood of receiving an antidepressant by 6% (OR 1.06, 95% CI: 1.01-1.11). CONCLUSION: Among adults with a comorbid cancer and depression diagnosis and a recorded U.S. ambulatory care visit in 2014-2015, 37% received antidepressant treatment. This suggests most patients with cancer and depression do not receive pharmacologic treatment for depression. Future studies are needed to investigate the impact of antidepressant treatment on health outcomes in this patient population.
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Depressão , Neoplasias , Adulto , Humanos , Feminino , Estados Unidos , Depressão/tratamento farmacológico , Depressão/epidemiologia , Depressão/diagnóstico , Estudos Transversais , Estudos Retrospectivos , Antidepressivos/uso terapêutico , Assistência Ambulatorial , Neoplasias/tratamento farmacológico , Neoplasias/epidemiologia , Padrões de Prática MédicaRESUMO
In oceanic ecosystems, the nature of barriers to gene flow and the processes by which populations may become isolated are different from the terrestrial environment, and less well understood. In this study we investigate a highly mobile species (the sperm whale, Physeter macrocephalus) that is genetically differentiated between an open North Atlantic population and the populations in the Mediterranean Sea. We apply high-resolution single nucleotide polymorphism (SNP) analysis to study the nature of barriers to gene flow in this system, assessing the putative boundary into the Mediterranean (Strait of Gibraltar and Alboran Sea region), and including novel analyses on structuring among sperm whale populations within the Mediterranean basin. Our data support a recent founding of the Mediterranean population, around the time of the last glacial maximum, and show concerted historical demographic profiles in both the Atlantic and the Mediterranean. In each region there is evidence for a population decline around the time of the founder event. The largest decline was seen within the Mediterranean Sea where effective population size is substantially lower (especially in the eastern basin). While differentiation is strongest at the Atlantic/Mediterranean boundary, there is also weaker but significant differentiation between the eastern and western basins of the Mediterranean Sea. We propose, however, that the mechanisms are different. While post-founding gene flow was reduced between the Mediterranean and Atlantic populations, within the Mediterranean an important factor differentiating the basins is probably a greater degree of admixture between the western basin and the North Atlantic and some level of isolation between the western and eastern Mediterranean basins. Subdivision within the Mediterranean Sea exacerbates conservation concerns and will require consideration of what distinct impacts may affect populations in the two basins.
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Ecossistema , Cachalote , Animais , Cachalote/genética , Mar Mediterrâneo , Genômica , Densidade Demográfica , Variação Genética/genéticaRESUMO
BACKGROUND: Corneal transparency may be compromised by viral infections causing corneal scarring, edema, and neovascularization. Ocular injury results from collateral damage induced by exacerbated immune response in corneal stroma. Myofibroblasts play a key role in this process by producing a disorganized extracellular matrix and inflammatory mediators. However, the immune response profile of myofibroblasts during viral infections is still under study. The aim of this work was to analyze the cytokine profile of human limbal myfibroblasts (HLMs) stimulated with the double-stranded RNA analog polyinosinic:polycytidylic acid (poly I:C) and to identify their signaling pathways. METHODS: HLMs were isolated from cadaveric sclera-corneal rims and stimulated with poly I:C (10 µg/ml) for 12 h. The secretion of 36 cytokines was measured using the Human Cytokine Array Panel A. The secretion of IFN-ß was quantified by ELISA. The expression of pattern recognition receptors (PRRs) such as TLR3, RIG-1 and MDA5 were analyzed by western blot assays. Furthermore, translocation of the nuclear factors NF-κB, IRF3, and IRF7 was assessed by fluorescence staining. In addition, the differentially expressed cytokines were analyzed using the Core Analysis Tool of the Ingenuity Pathway Analysis IPA software. RESULTS: HLMs stimulated with poly I:C increased (fold change > 2) the secretion of G-CSF, sTREM-1, CXCL1, CCL1, CXCL8, CXCL10, CXCL11, CCL2, CCL5, IL-13, IL-6, IL-1ra, and IFN-ß compared with HLMs under basal conditions. Poly I:C stimulation also induced the expression of RIG-1 (p < 0.001), but the expression of TLR3 and MDA5 was unmodified. Finally, HLMs increased nuclear translocation of NF-κB, IRF3, and IRF7 after poly I:C stimulation. Bioinformatic analysis identified canonical signaling pathways associated with cell adhesion and diapedesis, chemokine signaling, and activation of IRFs by cytosolic pattern recognition receptors. CONCLUSIONS: These results demonstrate that HLMs secrete cytokines involved in immune cell activation and chemotaxis. The data suggest a key role for HLMs during viral infections in cornea and extend our knowledge about the signaling pathways they trigger.
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NF-kappa B , Viroses , Antivirais/farmacologia , Córnea , Citocinas/metabolismo , Fator Estimulador de Colônias de Granulócitos/genética , Humanos , Interferon beta/metabolismo , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-13/genética , Interleucina-6/genética , Miofibroblastos/metabolismo , NF-kappa B/metabolismo , Poli I-C/farmacologia , RNA de Cadeia Dupla , Receptores de Reconhecimento de Padrão , Receptor 3 Toll-Like/metabolismoRESUMO
Reconstituted high-density lipoproteins (rHDLs) can transport and specifically release drugs and imaging agents, mediated by the Scavenger Receptor Type B1 (SR-B1) present in a wide variety of tumor cells, providing convenient platforms for developing theranostic systems. Usually, phospholipids or Apo-A1 lipoproteins on the particle surfaces are the motifs used to conjugate molecules for the multifunctional purposes of the rHDL nanoparticles. Cholesterol has been less addressed as a region to bind molecules or functional groups to the rHDL surface. To maximize the efficacy and improve the radiolabeling of rHDL theranostic systems, we synthesized compounds with bifunctional agents covalently linked to cholesterol. This strategy means that the radionuclide was bound to the surface, while the therapeutic agent was encapsulated in the lipophilic core. In this research, HYNIC-S-(CH2)3-S-Cholesterol and DOTA-benzene-p-SC-NH-(CH2)2-NH-Cholesterol derivatives were synthesized to prepare nanoparticles (NPs) of HYNIC-rHDL and DOTA-rHDL, which can subsequently be linked to radionuclides for SPECT/PET imaging or targeted radiotherapy. HYNIC is used to complexing 99mTc and DOTA for labeling molecules with 111, 113mIn, 67, 68Ga, 177Lu, 161Tb, 225Ac, and 64Cu, among others. In vitro studies showed that the NPs of HYNIC-rHDL and DOTA-rHDL maintain specific recognition by SR-B1 and the ability to internalize and release, in the cytosol of cancer cells, the molecules carried in their core. The biodistribution in mice showed a similar behavior between rHDL (without surface modification) and HYNIC-rHDL, while DOTA-rHDL exhibited a different biodistribution pattern due to the significant reduction in the lipophilicity of the modified cholesterol molecule. Both systems demonstrated characteristics for the development of suitable theranostic platforms for personalized cancer treatment.
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Nanopartículas , Medicina de Precisão , Animais , Camundongos , Distribuição Tecidual , Benzeno , Lipoproteínas HDL/metabolismo , Nanopartículas/uso terapêutico , Colesterol/metabolismo , Lipoproteínas/metabolismo , Radioisótopos , Fosfolipídeos , Receptores Depuradores/metabolismoRESUMO
Purpose: Viral infections such as herpetic keratitis (HSK) activate the innate immune response in the cornea triggering opacity and loss of vision. This condition is performed mainly by myofibroblasts that exacerbate secretion of inflammatory cytokines. Amniotic membrane transplantation (AMT) reduces ocular opacity and scarring inhibiting secretion of inflammatory cytokines and proliferation of myofibroblasts. We previously reported that the amniotic membrane (AM) favors an anti-inflammatory microenvironment inhibiting the secretion of inflammatory cytokines, expression of innate immune receptors, and translocation of nuclear NF-κB on human limbal myofibroblasts (HLMs). The aim of the present study was to determine whether the soluble factors of the AM decrease the immune response of HLMs stimulated with polyinosinic-polycytidylic acid sodium salt (poly I:C). Methods: The AM was incubated in Dulbecco's modified eagle medium (DMEM)/F12, and the supernatant was collected to obtain amniotic membrane conditioned medium (AMCM). HLMs were isolated from cadaveric sclera-corneal rims. HLMs were cultured in DMEM/F12 or AMCM and stimulated or not with poly I:C (10 µg/ml) for 12 h to analyze synthesis of CCL2, CCL5, CXCL10, MDA5, RIG-1, and TLR3 or for 2 h to analyze translocation of nuclear NF-kB, IRF3, and IRF7. The proteins contained on AMCM were analyzed by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and the acquired peptide ions were analyzed with the Mascot program using both National Center for Biotechnology Information (NCBI) and expressed sequence tag (EST) databases. Results: AMCM downregulated the mRNA levels of CCL2, CCL5, CXCL10, MDA5, RIG-1, and TLR3. In addition, AMCM decreased secretion of CCL2, CCL5, and CXCL10 and translocation of nuclear NF-κB. Interestingly, AMCM increased translocation of nuclear IRF3 and synthesis and secretion of type I IFN-ß. We also identified small leucine-rich proteoglycan lumican in the AMCM. The administration of rh-lumican to poly I:C-stimulated HLMs reduced the mRNA levels of CCL2, CCL5, and CXCL10. Conclusions: These results suggest that the AM can trigger an anti-inflammatory response on HLMs through soluble factors, and that lumican could play an important role in these effects.
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Âmnio/fisiologia , Meios de Cultivo Condicionados/farmacologia , Inflamação/prevenção & controle , Limbo da Córnea/citologia , Miofibroblastos/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunidade Inata/efeitos dos fármacos , Lumicana/farmacologia , Miofibroblastos/metabolismo , NF-kappa B/metabolismo , Fosforilação , Poli I-C/farmacologiaRESUMO
Sensorless speed estimation has been extensively studied for its use in control schemes. Nevertheless, it is also a key step when applying Motor Current Signature Analysis to induction motor diagnosis: accurate speed estimation is vital to locate fault harmonics, and prevent false positives and false negatives, as shown at the beginning of the paper through a real industrial case. Unfortunately, existing sensorless speed estimation techniques either do not provide enough precision for this purpose or have limited applicability. Currently, this is preventing Industry 4.0 from having a precise and automatic system to monitor the motor condition. Despite its importance, there is no research published reviewing this topic. To fill this gap, this paper investigates, from both theoretical background and an industrial application perspective, the reasons behind these problems. Therefore, the families of sensorless speed estimation techniques, mainly conceived for sensorless control, are here reviewed and thoroughly analyzed from the perspective of their use for diagnosis. Moreover, the algorithms implemented in the two leading commercial diagnostic devices are analyzed using real examples from a database of industrial measurements belonging to 79 induction motors. The analysis and discussion through the paper are synthesized to summarize the lacks and weaknesses of the industry application of these methods, which helps to highlight the open problems, challenges and research prospects, showing the direction in which research efforts have to be made to solve this important problem.
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Algoritmos , HumanosRESUMO
We present the case of a 41-year-old male with type A blood group and a diagnosis of fulminant liver failure of an unknown cause, who was admitted to the Intensive Care Unit (ICU) with severe coagulopathy (INR 6), hyperammonemia (250 umol/l) and grade IV encephalopathy (four King's criteria and both Clichy's criteria). He also had GCS 8/15, despite TDE-MARS, dialysis and ventilator support via endotracheal tube due to encephalopathy. He was registered as status 0 for liver transplant. In the absence of a compatible donor, the patient received an ABO incompatible liver transplant (LT), group O, using the piggyback technique, without any complications. He required transfusion of five units of plasma but did not require a blood transfusion.
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Falência Hepática Aguda , Transplante de Fígado , Sistema ABO de Grupos Sanguíneos , Adulto , Incompatibilidade de Grupos Sanguíneos , Transfusão de Sangue , Humanos , Falência Hepática Aguda/etiologia , Falência Hepática Aguda/cirurgia , MasculinoRESUMO
Induction motors are essential and widely used components in many industrial processes. Although these machines are very robust, they are prone to fail. Nowadays, it is a paramount task to obtain a reliable and accurate diagnosis of the electric motor health, so that a subsequent reduction of the required time and repairing costs can be achieved. The most common approaches to accomplish this task are based on the analysis of currents, which has some well-known drawbacks that may lead to false diagnosis. With the new developments in the technology of the sensors and signal processing field, the possibility of combining the information obtained from the analysis of different magnitudes should be explored, in order to achieve more reliable diagnostic conclusions, before the fault can develop into an irreversible damage. This paper proposes a smart-sensor that explores the weighted analysis of the axial, radial, and combination of both stray fluxes captured by a low-cost, easy setup, non-invasive, and compact triaxial stray flux sensor during the start-up transient through the short time Fourier transform (STFT) and characterizes specific patterns appearing on them using statistical parameters that feed a feature reduction linear discriminant analysis (LDA) and then a feed-forward neural network (FFNN) for classification purposes, opening the possibility of offering an on-site automatic fault diagnosis scheme. The obtained results show that the proposed smart-sensor is efficient for monitoring and diagnosing early induction motor electromechanical faults. This is validated with a laboratory induction motor test bench for individual and combined broken rotor bars and misalignment faults.
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Sustainable activated carbon can be obtained from the pyrolysis/activation of biomass wastes coming from different origins. Carbon obtained in this way shows interesting properties, such as high surface area, electrical conductivity, thermal and chemical stability, and porosity. These characteristics among others, such as a tailored pore size distribution and the possibility of functionalization, lead to an increased use of activated carbons in catalysis. The use of activated carbons from biomass origins is a step forward in the development of more sustainable processes enhancing material recycling and reuse in the frame of a circular economy. In this article, a perspective of different heterogeneous catalysts based on sustainable activated carbon from biomass origins will be analyzed focusing on their properties and catalytic performance for determined energy-related applications. In this way, the article aims to give the reader a scope of the potential of these tailor-made sustainable materials as a support in heterogeneous catalysis and future developments needed to improve catalyst performance. The selected applications are those related with H2 energy and the production of biomethane for energy through CO2 methanation.
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Biocombustíveis , Biomassa , Carvão Vegetal/química , Dióxido de Carbono/química , Catálise , Hidrogênio/química , Metano/químicaRESUMO
The interactions between prokaryotes and eukaryotes are abundant in nature. These microorganisms also interact in the human body. Fungal-bacteria interactions are present in many diseases. In this study, we evaluated the microbial interaction of Fusarium falciforme and Staphylococcus aureus developing mixed biofilm in vitro. When both microorganisms grew up together the mixed biofilm biomass decreased than F. falciforme monobiofilm biomass. S. aureus was able to interact and form aggregates over the mycelium and conidia surface of F. falciforme. Our results suggest that S. aureus could bind to colloidal chitin. On another hand, the supernatants from S. aureus biofilm and S. aureus-F. falciforme presented an antifungal effect over F. falciforme biofilm formation. Finally we found that the pH had an inhibitory effect over fungal biofilm formation. We concluded that S. aureus can affect the F. falciforme growth negatively in mixed biofilm involving factors like pH, supernatants compounds, anchor to chitin, and bacterial viability.
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Biofilmes/crescimento & desenvolvimento , Olho/microbiologia , Fusarium/crescimento & desenvolvimento , Interações Microbianas/fisiologia , Staphylococcus aureus/fisiologia , Ácido Acético , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Biomassa , Quitina , Fusarium/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio , Ácido Láctico , Viabilidade Microbiana/efeitos dos fármacos , Micélio , Esporos FúngicosRESUMO
INTRODUCTION/AIMS: In recent years, it has been shown that free fatty acids receptors (FFAR) of whose function in the cell surface plays a significant role in the regulation of cell function and nutrition as well are activated by various endogenous ligands, but mainly by fatty acids. Within FFAR of our interest are GPR 41, 43 and 120. The functions of these receptors are varied and dependent on the tissue where they are. The activation and signaling of these receptors, FFAR, are involved in many physiological processes, and currently the target of many drugs in metabolic disorders like obesity, diabetes and atherosclerosis. MATERIAL AND METHODS: Obesity was induced with hypercaloric diet (HD) in male Wistar rats for 20 weeks (n = 10). At the end, adipose tissue (abdominal and subcutaneous) was taken to perform assays for relative quantification mRNA expression by end-point RT-PCR and protein level expression by Western blot. RESULTS: These present data have shown for the first time that total mRNA isolation and protein expression from both adipose tissues (abdominal and subcutaneous) of rat in obesity condition yield significative statistical difference among the control versus obese groups, showing that the diet high in carbohydrates modifies the total presence of mRNA and protein level expression of the receptors GPR41, 43 and 120. CONCLUSIONS: Further comparative methods are in process to clarify whether or not the obesity changes the functional receptors in these two tissues for new pharmacological approaches.
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Obesidade/tratamento farmacológico , Obesidade/genética , Receptores Acoplados a Proteínas G/genética , Tecido Adiposo/metabolismo , Animais , Aterosclerose/tratamento farmacológico , Aterosclerose/patologia , Diabetes Mellitus/tratamento farmacológico , Diabetes Mellitus/patologia , Dieta Hiperlipídica , Modelos Animais de Doenças , Ácidos Graxos não Esterificados/genética , Ácidos Graxos não Esterificados/metabolismo , Regulação da Expressão Gênica/genética , Humanos , Insulina/genética , Insulina/metabolismo , Obesidade/metabolismo , Obesidade/patologia , Ratos , Receptores Acoplados a Proteínas G/metabolismoRESUMO
PURPOSE: To determine normative corneal densitometry values in relation to age, sex, refractive error, corneal thickness, and keratometry, measured using the Oculus Pentacam system. METHODS: Three hundred and thirty-eight healthy subjects (185 men; 153 women) with no corneal disease underwent an exhaustive ocular examination. Corneal densitometry was expressed in standardized grayscale units (GSU). RESULTS: The mean corneal densitometry over the total area was 16.46 ± 1.85 GSU. The Pearson correlation coefficient for total densitometry was r = 0.542 (p < 0.001). Statistically significant differences were found between men and women for the total area (p = 0.006), with readings of 16.22 ± 1.54 GSU and 16.60 ± 1.83 GSU, respectively. When the cornea was divided into layers of different depths, a significant correlation was found for all layers and age: r = 0.447 (p < 0.001), r = 0.563 (p < 0.001), and r = 0.520 (p < 0.001) for the anterior, central, and posterior layers, respectively. However, when the cornea was divided into concentric annuli starting from the center of the cornea, densitometry was strongly correlated only with age in the 6-10-mm annulus (p < 0.001). Neither mean keratometry nor spherical equivalent was correlated with corneal densitometry in any zone of the cornea (p > 0.05). CONCLUSIONS: This is the first report of normative corneal densitometry values in relation to keratometry, corneal thickness, and spherical equivalent measured with the latest Oculus Pentacam software. Corneal densitometry increases with age, but corneal keratometry and refractive parameters do not affect light scattering in the human cornea.
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Envelhecimento/fisiologia , Córnea/anatomia & histologia , Densitometria/métodos , Refração Ocular/fisiologia , Adulto , Paquimetria Corneana , Topografia da Córnea , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Corneal damage observed in a viral infection such as herpetic stromal keratitis is mainly caused by proinflammatory molecules released by resident cells in the response to viral antigens. There are pattern recognition receptors like MDA5, RIG-1, and TLR3, that recognize viral dsRNA and after activation, the innate immune response is exacerbated inducing the synthesis and secretion of inflammatory cytokines through NF-κB activation. Amniotic membrane (AM) has demonstrated to reduce inflammation by several mechanisms, however the effect of AM on innate immune receptors such as MDA5, RIG-1, and TLR3 has not been reported. In this study, we have determined that the presence of AM significantly inhibited the synthesis and secretion of proinflammatory cytokines on human limbal myofibroblasts (HLM) stimulated with poly I:C. Similarly, the presence of AM reduced the protein expression of MDA5, RIG-1, and TLR3 on poly I:C stimulated HLM. Additionally, the presence of the AM significantly inhibited the NF-κB nuclear translocation when the HLM were poly I:C stimulated, and concomitantly, the AM was able to relocate cadherins affecting the myofibroblastic cellular morphology. These results suggest that AM generates an anti-inflammatory microenvironment, and specific inhibition of NFκB nuclear translocation on infected corneal tissue would reduce the inflammation undesirable effects, explaining in part the beneficial usefulness of transplanting AM on herpetic stromal keratitis.
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Âmnio/fisiologia , Imunidade Inata/fisiologia , Limbo da Córnea/citologia , Miofibroblastos/metabolismo , NF-kappa B/metabolismo , Receptores de Reconhecimento de Padrão/metabolismo , Células Cultivadas , Citocinas/metabolismo , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Miofibroblastos/efeitos dos fármacos , Poli I-C/farmacologia , Transporte ProteicoRESUMO
The use of environmental DNA (eDNA) to assess the presence of biological communities has emerged as a promising monitoring tool in the marine conservation landscape. Moreover, advances in Next-Generation Sequencing techniques, such as DNA metabarcoding, enable multi-species detection in mixed samples, allowing the study of complex ecosystems such as oceanic ones. We aimed at using these molecular-based techniques to characterize cetacean communities, as well as potential prey on the northern coast of Mainland Portugal. During four seasonal campaigns (summer 2021 to winter 2022/2023), seawater samples were collected along with visual records of cetacean occurrence. The eDNA isolated from 64 environmental samples was sequenced in an Illumina platform, with universal primers targeting marine vertebrates. Five cetacean species were identified by molecular detection: common dolphin (Delphinus delphis), bottlenose dolphin (Tursiops truncatus), Risso's dolphin (Grampus griseus), harbor porpoise (Phocoena phocoena) and fin whale (Balaenoptera physalus). Overall, except for the latter (not sighted during the campaigns), this cetacean community composition was similar to that obtained through visual monitoring, and the complementary results suggest their presence in the region all year round. In addition, the positive molecular detections of Balaenoptera physalus are of special relevance since there are no records of this species reported on scientific bibliography in the area. The detection of multiple known prey of the identified dolphins indicates an overlap between predator and prey in the study area, which suggests that these animals may use this coastal area for feeding purposes. While this methodological approach remains in a development stage, the present work highlights the benefits of using eDNA to study marine communities, with specific applications for research on cetacean distribution and feeding ecology.
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Biodiversidade , Cetáceos , DNA Ambiental , Animais , DNA Ambiental/análise , DNA Ambiental/genética , Cetáceos/genética , Código de Barras de DNA Taxonômico/métodos , Portugal , Ecossistema , Comportamento Alimentar , Monitoramento Ambiental/métodos , Estações do Ano , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
Systems that are connected through multiple hydrogen bonds are the cornerstone of molecular recognition processes in biology, and they are increasingly being employed in supramolecular chemistry, specifically in molecular self-assembly processes. For this reason, the effects of different substituents (NO2, CN, F, Cl, Br, OCH3 and NH2) on the electronic structure, and consequently on the magnitude of hydrogen bonds in triple AAA-DDD arrays (A=acceptor, D=donor) were evaluated in the light of topological [electron localization function (ELF) and quantum theory of atoms in molecules (QTAIM)], energetic [Su-Li energy-decomposition analysis (EDA) and natural bond orbital analysis (NBO)], and geometrical analysis. The results based on local H-bond descriptors (geometries, QTAIM, ELF, and NBO) indicate that substitutions with electron-withdrawing groups on the AAA module tend to strengthen, whereas electron-donating substituents tend to weaken the covalent character of the AAA-DDD intermolecular H-bonds, and also indicate that the magnitude of the effect is dependent on the position of substitution. In contrast, Su-Li EDA results show an opposite behavior when compared to local H-bond descriptors, indicating that electron-donating substituents tend to increase the magnitude of H-bonds in AAA-DDD arrays, and thus suggesting that the use of local H-bond descriptors describes the nature of H bonds only partially, not providing enough insight about the strength of such H bonds.
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Aldeídos/química , Di-Hidropiridinas/química , Naftiridinas/química , Hidrogênio/química , Ligação de Hidrogênio , Modelos MolecularesRESUMO
In the pygmy sperm whale (Kogia breviceps, Blainville 1838), vibrissae are present in neonates, but within a few months the hairs are lost, and the structures remain as empty vibrissal crypts (VCs). In this work, we have studied histologically the facial vibrissal follicles of two juveniles and one adult specimens stranded dead. A few VCs with no visible hairs were found grouped in a row rostral to each eye. The follicular lumen, covered by a simple squamous epithelium, showed invaginations in the most superficial part. Beneath the epithelium, the follicle walls were made of loose connective tissue and were encircled by a thick capsule of dense connective tissue. In juveniles, a dermal papilla was found basally and, from it, a non-keratinized pseudo hair grew upwards but did not reach the skin surface. The VCs were richly innervated and irrigated. Many lamellated corpuscles were identified in the subluminal connective tissue of the crypt walls. A large venous cavernous plexus was located beneath and around the hair papilla. The main differences observed in the adult specimen were the degeneration and calcification of both the dermal papilla and the pseudo hair, and the absence of the venous cavernous plexus, albeit maintaining a rich vascularization and innervation. Our study revealed that VCs of the pygmy sperm whale possess features of fully functional sensory structures, with a microanatomy different from those described in other species. In addition, they undergo a postnatal morphological transformation, which implies functional differences between the VCs of neonates and adults.
Assuntos
Folículo Piloso , Baleias , Animais , Cabelo , Face , Couro CabeludoRESUMO
The Iberian harbour porpoise population is small and fisheries bycatch has been described as one of its most important threats. Data on harbour porpoise strandings collected by the Portuguese and Galician stranding networks between 2000 and 2020 are indicative of a recent mortality increase in the western Iberian coast (particularly in northern Portugal). Overall, in Portugal and Galicia, individuals stranded due to confirmed fishery interaction represented 46.98% of all analysed porpoises, and individuals stranded due to probable fishery interaction represented another 10.99% of all analysed porpoises. Considering the Portuguese annual abundance estimates available between 2011 and 2015, it was possible to calculate that an annual average of 207 individuals was removed from the population in Portuguese waters alone, which largely surpasses the potential biological removal (PBR) estimates (22 porpoises, CI: 12-43) for the same period. These results are conservative and bycatch values from strandings are likely underestimated. A structured action plan accounting for new activities at sea is needed to limit the Iberian porpoise population decline. Meanwhile, there is an urgent need for a fishing effort reorganization to directly decrease porpoise mortality.