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Calcineurin, or protein phosphatase 2B (PP2B), the Ca2+ and calmodulin-activated phosphatase and target of immunosuppressants, has many substrates and functions that remain uncharacterized. By combining rapid proximity-dependent labeling with cell cycle synchronization, we mapped the spatial distribution of calcineurin in different cell cycle stages. While calcineurin-proximal proteins did not vary significantly between interphase and mitosis, calcineurin consistently associated with multiple centrosomal and/or ciliary proteins. These include POC5, which binds centrins in a Ca2+-dependent manner and is a component of the luminal scaffold that stabilizes centrioles. We show that POC5 contains a calcineurin substrate motif (PxIxIT type) that mediates calcineurin binding in vivo and in vitro. Using indirect immunofluorescence and ultrastructure expansion microscopy, we demonstrate that calcineurin colocalizes with POC5 at the centriole, and further show that calcineurin inhibitors alter POC5 distribution within the centriole lumen. Our discovery that calcineurin directly associates with centriolar proteins highlights a role for Ca2+ and calcineurin signaling at these organelles. Calcineurin inhibition promotes elongation of primary cilia without affecting ciliogenesis. Thus, Ca2+ signaling within cilia includes previously unknown functions for calcineurin in maintenance of cilia length, a process that is frequently disrupted in ciliopathies.
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Calcineurina , Cílios , Calcineurina/metabolismo , Cílios/metabolismo , Cálcio/metabolismo , Centrossomo/metabolismo , Centríolos/metabolismo , Proteínas/metabolismoRESUMO
Lung adenocarcinoma (LUAD) is characterized by a high incidence rate and mortality. Recently, POC1 centriolar protein A (POC1A) has emerged as a potential biomarker for various cancers, contributing to cancer onset and development. However, the association between POC1A and LUAD remains unexplored. We extracted The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) data sets to analyse the differential expression of POC1A and its relationship with clinical stage. Additionally, we performed diagnostic receiver operator characteristic (ROC) curve analysis and Kaplan-Meier (KM) survival analysis to assess the diagnostic and prognostic value of POC1A in LUAD. Furthermore, we investigated the correlation between POC1A expression and immune infiltration, tumour mutation burden (TMB), immune checkpoint expression and drug sensitivity. Finally, we verified POC1A expression using real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC). Cell experiments were conducted to validate the effect of POC1A expression on the proliferation, migration and invasion of lung cancer cells. POC1A exhibited overexpression in most tumour tissues, and its overexpression in LUAD was significantly correlated with late-stage presentation and poor prognosis. The high POC1A expression group showed lower levels of immune infiltration but higher levels of immune checkpoint expression and TMB. Moreover, the high POC1A expression group demonstrated sensitivity to multiple drugs. In vitro experiments confirmed that POC1A knockdown led to decreased proliferation, migration, and invasion of lung cancer cells. Our findings suggest that POC1A may contribute to tumour development by modulating the cell cycle and immune cell infiltration. It also represents a potential therapeutic target and marker for the diagnosis and prognosis of LUAD.
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Adenocarcinoma de Pulmão , Neoplasias Pulmonares , Humanos , Adenocarcinoma de Pulmão/genética , Ciclo Celular , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Divisão Celular , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Neoplasias Pulmonares/genética , Regulação para Cima/genéticaRESUMO
Subjective cognitive decline (SCD) is a high-risk population in the preclinical stage of Alzheimer's disease (AD), and olfactory dysfunction is a risk factor for dementia progression. The present study aimed to explore the patterns of functional connectivity (FC) changes in the olfactory neural circuits during olfactory stimulation in SCD subjects. A total of 56 SCD subjects and 56 normal controls (NCs) were included. All subjects were assessed with a cognitive scale, an olfactory behavior test, and olfactory task-based functional magnetic resonance imaging scanning. The FC differences in olfactory neural circuits between the two groups were analyzed by the generalized psychophysiological interaction. Additionally, we calculated and compared the activation of brain regions within the olfactory neural circuits during odor stimulation, the volumetric differences in brain regions showing FC differences between groups, and the correlations between neuroimaging indicators and olfactory behavioral and cognitive scale scores. During odor stimulation, the FC between the bilateral primary olfactory cortex (bPOC) and the right hippocampus in the SCD group was significantly reduced; while the FC between the right hippocampus and the right frontal cortex was significantly increased in the SCD group. The bPOC of all subjects showed significant activation, but no significant difference in activation between groups was found. No significant differences were observed in the volume of the brain regions within the olfactory neural circuits or in olfactory behavior between groups. The volume of the bPOC and right frontal cortex was significantly positively correlated with olfactory identification, and the volume of the right frontal cortex and right hippocampus was significantly correlated with cognitive functions. Furthermore, a significant correlation between the activation of bPOC and the olfactory threshold was found in the whole cohort. These results suggested that while the structure of the olfactory neural circuits and olfactory behavior in SCD subjects remained stable, there were significant changes observed in the FC of the olfactory neural circuits (specifically, the POC-hippocampus-frontal cortex neural circuits) during odor stimulation. These findings highlight the potential of FC alterations as sensitive imaging markers for identifying high-risk individuals in the early stage of AD.
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Disfunção Cognitiva , Lobo Frontal , Hipocampo , Imageamento por Ressonância Magnética , Córtex Olfatório , Humanos , Masculino , Feminino , Idoso , Pessoa de Meia-Idade , Disfunção Cognitiva/diagnóstico por imagem , Disfunção Cognitiva/fisiopatologia , Disfunção Cognitiva/etiologia , Córtex Olfatório/diagnóstico por imagem , Córtex Olfatório/fisiologia , Córtex Olfatório/fisiopatologia , Hipocampo/diagnóstico por imagem , Hipocampo/fisiopatologia , Lobo Frontal/diagnóstico por imagem , Lobo Frontal/fisiopatologia , Percepção Olfatória/fisiologia , Rede Nervosa/diagnóstico por imagem , Rede Nervosa/fisiopatologia , Rede Nervosa/fisiologia , Conectoma , OdorantesRESUMO
Peatlands are globally important stores of soil carbon (C) formed over millennial timescales but are at risk of destabilization by human and climate disturbance. Pools are ubiquitous features of many peatlands and can contain very high concentrations of C mobilized in dissolved and particulate organic form and as the greenhouses gases carbon dioxide (CO2 ) and methane (CH4 ). The radiocarbon content (14 C) of these aquatic C forms tells us whether pool C is generated by contemporary primary production or from destabilized C released from deep peat layers where it was previously stored for millennia. We present novel 14 C and stable C (δ13 C) isotope data from 97 aquatic samples across six peatland pool locations in the United Kingdom with a focus on dissolved and particulate organic C and dissolved CO2 . Our observations cover two distinct pool types: natural peatland pools and those formed by ditch blocking efforts to rewet peatlands (restoration pools). The pools were dominated by contemporary C, with the majority of C (~50%-75%) in all forms being younger than 300 years old. Both pool types readily transform and decompose organic C in the water column and emit CO2 to the atmosphere, though mixing with the atmosphere and subsequent CO2 emissions was more evident in natural pools. Our results show little evidence of destabilization of deep, old C in natural or restoration pools, despite the presence of substantial millennial-aged C in the surrounding peat. One possible exception is CH4 ebullition (bubbling), with our observations showing that millennial-aged C can be emitted from peatland pools via this pathway. Our results suggest that restoration pools formed by ditch blocking are effective at preventing the release of deep, old C from rewetted peatlands via aquatic export.
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Dióxido de Carbono , Gases de Efeito Estufa , Humanos , Idoso , Dióxido de Carbono/análise , Ciclo do Carbono , Solo , Mudança ClimáticaRESUMO
SOFT syndrome (Short stature-Onychodysplasia-Facial dysmorphism-hypoTrichosis) is a rare primordial dwarfism syndrome caused by biallelic variants in POC1A encoding a centriolar protein. To refine the phenotypic spectrum of SOFT syndrome, recently shown to include metabolic features, we conducted a systematic review of all published cases (19 studies, including 42 patients). The SOFT tetrad affected only 24 patients (57%), while all cases presented with short stature from birth (median height: -5.5SDS([-8.5]-[-2.8])/adult height: 132.5 cm(103.5-148)), which was most often disproportionate (90.5%), with relative macrocephaly. Bone involvement resulted in short hands and feet (100%), brachydactyly (92.5%), metaphyseal (92%) or epiphyseal (84%) anomalies, and/or sacrum/pelvis hypoplasia (58%). Serum IGF-I was increased (median IGF-I level: + 2 SDS ([-0.5]-[+ 3])). Recombinant human growth hormone (rhGH) therapy was stopped for absence/poor growth response (7/9 patients, 78%) and/or hyperglycemia (4/9 patients, 45%). Among 11 patients evaluated, 10 (91%) presented with central distribution of fat (73%), clinical (64%) and/or biological insulin resistance (IR) (100%, median HOMA-IR: 18), dyslipidemia (80%), and hepatic steatosis (100%). Glucose tolerance abnormalities affected 58% of patients aged over 10 years. Patients harbored biallelic missense (52.4%) or truncating (45.2%) POC1A variants. Biallelic null variants, affecting 36% of patients, were less frequently associated with the SOFT tetrad (33% vs 70% respectively, p = 0.027) as compared to other variants, without difference in the prevalence of metabolic abnormalities. POC1A should be sequenced in children with short stature, altered glucose/insulin homeostasis and/or centripetal fat distribution. In patients with SOFT syndrome, rhGH treatment is not indicated, and IR-related complications should be regularly screened and monitored.PROSPERO registration: CRD42023460876.
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Cardiovascular diseases (CVD) are a range of diseases, pointing the functional hindrances in the heart and blood vessels of the human system that contributes to 48.6 % of the world's adult death rate. The diagnosis of CVD relies upon the Electro Cardio Gram (ECG) and detection of muscle markers such as troponins. Among the cardiac trio, Cardiac Troponin I (cTnI) weighing 23 KiloDalton (kDa) is a sorted biomarker for CVD. cTnI remains high in the blood after 1-2 weeks of myocardial damage. Testing of cTnI in CVD patients aids in diagnosis and risk stratification of the disease. Different determination systems including optical, electrochemical, and acoustic have been put forward for monitoring the cTnI which are Point of Care (POC) that promotes simple and sensitive detection of cTnI. The modern era has paved way to high-sensitivity Troponin I (hscTnI) devices that can detect up to 0.01 ng/ml in human blood/plasma/serum. Yet, the practice of hscTnI is impracticable due to cost inefficiency. Development of new hscTnI devices with minimal investment and maximal detection range will meet the global requirement. This review gives an over view on different detection systems of cardiac troponin I which stands as a translational detection molecule for CVDs.
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Doenças Cardiovasculares , Troponina I , Adulto , Humanos , Troponina T , Relevância Clínica , BiomarcadoresRESUMO
Low-molecular drug discovery using DNA-encoded chemical library (DEL) is a powerful technology, although improving the partitioning efficiency of affinity ligands from DEL remains a challenge. Here, we assessed the usefulness of microbead-assisted capillary electrophoresis (MACE) for partitioning peptide-oligonucleotide conjugates (POCs), in which high selection pressure is applied because of different mobility of target-modified beads and POCs during CE. Despite their different charge characteristics, all POCs were well separated from the beads. When bead extraction was performed, the tagged DNA amplification was observed only in the couple of a ligand/target, suggesting proficiently specific partitioning of peptide ligands was accomplished using MACE.
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Oligonucleotídeos , Peptídeos , Microesferas , Peptídeos/química , Oligonucleotídeos/química , Eletroforese Capilar/métodos , DNA/química , DNA de Cadeia SimplesRESUMO
BACKGROUND: A life-threatening anaphylactic shock can occur if a patient with undiagnosed immunoglobulin A (IgA) deficiency (i.e., IgA levels <500 ng/mL) receives IgA-containing blood, hence the need for a rapid, point-of-care (POC) method for IgA deficiency screening. Enzyme-linked immunosorbent assay (ELISA) is routinely used to detect IgA, but this method requires trained specialists and ≥24 h to obtain a result. We developed a surface plasmon resonance (SPR)-based protocol to identify IgA-deficient patients or donors within 1 h. MATERIALS AND METHODS: The SPR sensor relies on the detection of IgAs captured by primary antibodies adsorbed on the SPR chip and quantified with secondary antibodies. The sensor was calibrated from 0 to 2000 ng/mL in buffer, IgA-depleted human serum, and plasma samples from IgA-deficient individuals. A critical concentration of 500 ng/mL was set for IgA deficiency. The optimized sensor was then tested on eight plasma samples with known IgA status (determined by ELISA), including five with IgA deficiency and three with normal IgA levels. RESULTS: The limit of detection was estimated at 30 ng/mL in buffer and 400 ng/mL in diluted plasma. The results obtained fully agreed with ELISA among the eight plasma samples tested. The protocol distinguished IgA-deficient from normal samples, even for samples with an IgA concentration closer to critical concentration. DISCUSSION: In conclusion, we developed a reliable POC assay for the quantification of IgA in plasma. This test may permit POC testing at blood drives and centralized centers to maintain reserves of IgA-deficient blood and in-hospital testing of blood recipients.
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Deficiência de IgA , Imunoglobulina A , Ressonância de Plasmônio de Superfície , Humanos , Ressonância de Plasmônio de Superfície/métodos , Ressonância de Plasmônio de Superfície/instrumentação , Imunoglobulina A/sangue , Deficiência de IgA/sangue , Deficiência de IgA/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodosRESUMO
OBJECTIVE: To evaluate cell-free DNA (cfDNA) testing as a non-invasive approach to detecting aneuploidies in clinical miscarriages. DESIGN: A retrospective cohort study of women with pregnancy loss. SETTING: Hospitals and genetic analysis laboratories. POPULATION OR SAMPLE: Pregnancy losses in the period 2021-2022. METHODS: Results derived from non-invasive cfDNA testing (Veriseq NIPT Solution V2) of maternal blood and invasive analysis of products of conception (POC) (Ion ReproSeq) compared in 120 women who suffered a miscarriage. MAIN OUTCOME MEASURES: Concordance rate results, cfDNA testing performance, non-informative rate (NIR) and fetal fraction (FF). RESULTS: We found no significant differences in the NIR between invasive (iPOC) and non-invasive (niPOC) analysis of POC (10.0% [12/120] versus 16.7% [20/120]). Of 120 samples, 90 provided an informative result in iPOC and niPOC groups (75%). cfDNA analysis correctly identified 74/87 (85.1%) samples (excluding triploidies). Sensitivity and specificity were 79.4% and 100%, respectively; all discordant cases were female. A binomial logistic model suggested fetal sex as the only variable influencing the concordance rate (P = 0.035). A Y-chromosome-based FF estimate allowed the optimal reclassification of cfDNA of non-informative male fetuses and a more accurate evaluation of cfDNA testing performance. The difference between the two FF estimates (native algorithm and Y-chromosome-based) suggests that female non-concordant cases may represent non-informative cases. CONCLUSIONS: Cell-free DNA-based testing provides a non-invasive approach to determining the genetic cause of clinical miscarriage.
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Aborto Espontâneo , Ácidos Nucleicos Livres , Gravidez , Feminino , Masculino , Humanos , Aborto Espontâneo/diagnóstico , Aborto Espontâneo/genética , Estudos Retrospectivos , Diagnóstico Pré-Natal/métodos , Aneuploidia , TrissomiaRESUMO
This study presents a groundbreaking approach for the early detection of chronic kidney disease (CKD) and other urological disorders through an image-label-free, multi-dipstick identification method, eliminating the need for complex machinery, label libraries, or preset coordinates. Our research successfully identified reaction pads on 187 multi-dipsticks, each with 11 pads, leveraging machine learning algorithms trained on human urine data. This technique aims to surpass traditional colourimetric methods and concentration-colour curve fitting, offering more robust and precise community screening and home monitoring capabilities. The developed algorithms enhance the generalizability of machine learning models by extracting primary colours and correcting urine colours on each reaction pad. This method's cost-effectiveness and portability are significant, as it requires no additional equipment beyond a standard smartphone. The system's performance rivals professional medical equipment without auxiliary lighting or flash under regular indoor light conditions, effectively managing false positives and negatives across various categories with remarkable accuracy. In a controlled experimental setting, we found that random forest algorithms, based on a Bagging strategy and applied in the HSV colour space, showed optimal results in smartphone-assisted urinalysis. This study also introduces a novel urine colour correction method, significantly improving machine learning model performance. Additionally, ISO parameters were identified as crucial factors influencing the accuracy of smartphone-based urinalysis in the absence of additional lighting or optical configurations, highlighting the potential of this technology in low-resource settings.
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Insuficiência Renal Crônica , Smartphone , Humanos , Urinálise/métodos , Algoritmos , Aprendizado de MáquinaRESUMO
Airborne animal viral pathogens can rapidly spread and become a global threat, resulting in substantial socioeconomic and health consequences. To prevent and control potential epidemic outbreaks, accurate, fast, and affordable point-of-care (POC) tests are essential. As a proof-of-concept, we have developed a molecular system based on the loop-mediated isothermal amplification (LAMP) technique for avian metapneumovirus (aMPV) detection, an airborne communicable agent mainly infecting turkeys and chickens. For this purpose, a colorimetric system was obtained by coupling the LAMP technique with specific DNA-functionalized AuNPs (gold nanoparticles). The system was validated using 50 different samples (pharyngeal swabs and tracheal tissue) collected from aMPV-infected and non-infected chickens and turkeys. Viral detection can be achieved in about 60 min with the naked eye, with 100% specificity and 87.88% sensitivity for aMPV. In summary, this novel molecular detection system allows suitable virus testing in the field, with accuracy and limit of detection (LOD) values highly close to qRT-PCR-based diagnosis. Furthermore, this system can be easily scalable to a platform for the detection of other viruses, addressing the current gap in the availability of POC tests for viral detection in poultry farming. KEY POINTS: â¢aMPV diagnosis using RT-LAMP is achieved with high sensitivity and specificity. â¢Fifty field samples have been visualized using DNA-nanoprobe validation. â¢The developed system is a reliable, fast, and cost-effective option for POCT.
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Galinhas , Ouro , Metapneumovirus , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Infecções por Paramyxoviridae , Doenças das Aves Domésticas , Sensibilidade e Especificidade , Metapneumovirus/genética , Metapneumovirus/isolamento & purificação , Animais , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/economia , Galinhas/virologia , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/economia , Infecções por Paramyxoviridae/diagnóstico , Infecções por Paramyxoviridae/veterinária , Infecções por Paramyxoviridae/virologia , Doenças das Aves Domésticas/virologia , Doenças das Aves Domésticas/diagnóstico , Ouro/química , Perus , Nanopartículas Metálicas/química , Limite de Detecção , Colorimetria/métodos , DNA Viral/genéticaRESUMO
We report a 2.2 year-old-boy, born of consanguineous marriage, referred for short stature, with history of neonatal death and skeletal deformities in his older sibling. Rhizo-mesomelic dwarfism was detected antenatally. Within 24 hours of birth, he developed multiple seizures. Examination revealed severe short stature, dolichocephaly, broad forehead, deep set eyes, low set ears, bulbous nose, small, irregular teeth, pointed chin, and triangular facies. He had rhizomelic shortening, stubby fingers, pes planus, and scanty hair. Neurological evaluation revealed ataxia, hypotonia, and global developmental delay. Skeletal survey radiograph revealed shallow acetabuli, short femurs and humerus, short, broad metacarpals and short cone-shaped phalanges with cupping of phalangeal bases. Clinical exome analysis revealed homozygous mutations involving the POC1A gene and the SLC13A5 gene responsible for SOFT syndrome and Kohlschutter-Tonz syndrome respectively, which were inherited from the parents. Both these syndromes are extremely rare, and their co-occurrence is being reported for the first time.
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Anormalidades Múltiplas , Amelogênese Imperfeita , Demência , Nanismo , Epilepsia , Osteocondrodisplasias , Simportadores , Masculino , Recém-Nascido , Humanos , Pré-Escolar , Amelogênese Imperfeita/genética , Anormalidades Múltiplas/genética , Osteocondrodisplasias/genética , Nanismo/genética , Nanismo/diagnóstico , Proteínas do Citoesqueleto , Proteínas de Ciclo CelularRESUMO
Salivary pH serves as a valuable and useful diagnostic marker for periodontal disease, as it not only plays a critical role in disease prevention but also in its development. Typically, saliva sampling is collected by draining and spitting it into collection tubes or using swabs. In this study, we have developed a Point-of-Care (POC) device for in situ determination of oral pH without the need for complex instruments, relying solely on a smartphone as the detection device. Our system utilizes a non-toxic vegetable colourimetric indicator, immobilized on a chitosan membrane located on a disposable stick, enabling direct sampling within the buccal cavity. An ad hoc designed 3D-printed attachment is used to ensure accurate positioning and alignment of the stick, as well as isolation from external lighting conditions. A custom-developed smartphone application captures and automatically processes the image of the sensing membrane, providing the salivary pH results. After optimizing the cocktail composition, the developed sensors demonstrated the capacity to determine pH within a range of 5.4 to 8.1 with a remarkable precision of 0.6%, achieving a very short analysis time of just 1 min. A stability study conducted on the sensing membranes revealed a lifetime of 50 days. To validate the performance of our analytical device, we compared its results against those obtained from a calibrated pH-meter, using a group of individuals. The device exhibited an average error of 2.4% when compared with the pH-meter results, confirming its reliability and accuracy.
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Quitosana , Smartphone , Humanos , Sistemas Automatizados de Assistência Junto ao Leito , Reprodutibilidade dos Testes , Concentração de Íons de HidrogênioRESUMO
Acrylamide (AA), an odorless and colorless organic small-molecule compound found generally in thermally processed foods, possesses potential carcinogenic, neurotoxic, reproductive, and developmental toxicity. Compared with conventional methods for AA detection, bio/chemical sensors have attracted much interest in recent years owing to their reliability, sensitivity, selectivity, convenience, and low cost. This paper provides a comprehensive review of bio/chemical sensors utilized for the detection of AA over the past decade. Specifically, the content is concluded and systematically organized from the perspective of the sensing mechanism, state of selectivity, linear range, detection limits, and robustness. Subsequently, an analysis of the strengths and limitations of diverse analytical technologies ensues, contributing to a thorough discussion about the potential developments in point-of-care (POC) for AA detection in thermally processed foods at the conclusion of this review.
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Acrilamida , Técnicas Biossensoriais , Sistemas Automatizados de Assistência Junto ao Leito , Acrilamida/análise , Acrilamida/química , Técnicas Biossensoriais/métodos , Humanos , Análise de Alimentos/métodos , Contaminação de Alimentos/análiseRESUMO
Nucleic acid tests are key tools for the detection and diagnosis of many diseases. In many cases, the amplification of the nucleic acids is required to reach a detectable level. To make nucleic acid amplification tests more accessible to a point-of-care (POC) setting, isothermal amplification can be performed with a simple heating source. Although these tests are being performed in bulk reactions, the quantification is not as accurate as it would be with digital amplification. Here, we introduce the use of the vibrating sharp-tip capillary for a simple and portable system for tunable on-demand droplet generation. Because of the large range of droplet sizes possible and the tunability of the vibrating sharp-tip capillary, a high dynamic range (~2 to 6000 copies/µL) digital droplet loop-mediated isothermal amplification (ddLAMP) system has been developed. It was also noted that by changing the type of capillary on the vibrating sharp-tip capillary, the same mechanism can be used for simple and portable DNA fragmentation. With the incorporation of these elements, the present work paves the way for achieving digital nucleic acid tests in a POC setting with limited resources.
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Técnicas de Amplificação de Ácido Nucleico , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/instrumentação , Vibração , Sistemas Automatizados de Assistência Junto ao Leito , Humanos , Ácidos Nucleicos/análise , DNA/análise , DNA/genética , DNA/químicaRESUMO
PURPOSE: Limited research exists on translation of in-vitro glucose measurement interfering compounds to the in-vivo situation. We investigated whether Point-of-Care glucose measurements by Accu Chek Inform II (ACI II) were accurate to monitor glucose concentrations during surgery with general anesthesia by comparing with the reference laboratory hexokinase plasma glucose test. METHOD: Patients undergoing surgery with general anesthesia were included. Anesthesia was maintained with either Sevoflurane or Total intravenous anesthesia (TIVA). Prior to and after induction, blood glucose was measured with ACI II and the hexokinase test. Bland-Altman analysis was performed to assess method agreement. Subgroup analyses on glucose measurement differences per type of maintenance anesthesia were performed. RESULTS: Thirty-nine patients were included, and 78 measurements were performed. All paired measurements had clinically acceptable agreement with a percentage error of 10.0% (95% CI 8.0 to 11.9). The mean difference (95% limits of agreement) between ACI II and hexokinase for all measurements was 0.0 mmol/L (-0.7 to 0.7 mmol/L). Before induction (n = 39), mean difference was -0.1 mmol/L (-0.6 to 0.4 mmol/L), and after induction (n = 39), mean difference was 0.1 mmol/L (-0.8 to 0.9 mmol/L). Further investigation showed the difference varied per test for patients receiving Sevoflurane compared to patients receiving TIVA (-0.2 ± 0.4 mmol/L vs. 0.4 ± 0.3 mmol/L, p < 0.001). Before and after induction, the difference between ACI II and hexokinase measurements increased for patients receiving Sevoflurane compared to patients receiving TIVA (0.4 ± 0.4 mmol/L vs. -0.4 ± 0.3 mmol/L, p < 0.001). CONCLUSION: The agreement between glucose measurements using ACI II and the reference laboratory hexokinase test was clinically acceptable with a percentage error of 10.0% (95% CI 8.0 to 11.9). The use of TIVA may negatively affect the measurement performance of the ACI II.
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Anestésicos Inalatórios , Propofol , Humanos , Sevoflurano , Hexoquinase , Anestesia Geral , Glicemia/análise , Estudos de Coortes , Anestesia Intravenosa , Anestésicos IntravenososRESUMO
The rapid and accurate diagnosis of meningitis is critical for preventing severe complications and fatalities. This study addresses the need for accessible diagnostics in the absence of specialized equipment by developing a novel diagnostic assay. The assay utilizes dual-priming isothermal amplification (DAMP) with unique internal primers to significantly reduce non-specificity. For fluorescence detection, the dye was selected among Brilliant Green, Thioflavin T, and dsGreen. Brilliant Green is preferred for this assay due to its availability, high fluorescence level, and optimal sample-to-background (S/B) ratio. The assay was developed for the detection of the primary causative agents of meningitis (Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae), and tested on clinical samples. The developed method demonstrated high specificity, no false positives, sensitivity comparable to that of loop-mediated isothermal amplification (LAMP), and a high S/B ratio. This versatile assay can be utilized as a standalone test or an integrated assay into point-of-care systems for rapid and reliable pathogen detection.
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Haemophilus influenzae , Meningites Bacterianas , Técnicas de Diagnóstico Molecular , Neisseria meningitidis , Técnicas de Amplificação de Ácido Nucleico , Streptococcus pneumoniae , Neisseria meningitidis/genética , Neisseria meningitidis/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/isolamento & purificação , Humanos , Haemophilus influenzae/genética , Haemophilus influenzae/isolamento & purificação , Meningites Bacterianas/diagnóstico , Meningites Bacterianas/microbiologia , Técnicas de Diagnóstico Molecular/métodos , Sensibilidade e EspecificidadeRESUMO
Soil microorganisms are the drivers of soil organic carbon (SOC) mineralization, and the activities of these microorganisms are considered to play a key role in SOC dynamics. However, studies of the relationship between soil microbial carbon metabolism and SOC stocks are rare, especially in different physical fractions (e.g., particulate organic carbon (POC) fraction and mineral-associated organic carbon (MAOC) fraction). In this study, we investigated the changing patterns of SOC stocks, POC stocks, MAOC stocks and microbial carbon metabolism (e.g., microbial growth, carbon use efficiency and biomass turnover time) at 0-20 cm along an elevational gradient in a subtropical mountain forest ecosystem. Our results showed that SOC and POC stocks increased but MAOC stocks remained stable along the elevational gradient. Soil microbial growth increased while microbial turnover time decreased with elevation. Using structural equation modeling, we found that heightened microbial growth is associated with elevated POC stocks. Moreover, MAOC stocks positively correlate with microbial growth but show negative associations with both POC stocks and soil pH. Overall, the increase in SOC stocks along the elevational gradient is primarily driven by changes in POC stocks rather than MAOC stocks. These findings underscore the importance of considering diverse soil carbon fractions and microbial activities in predicting SOC responses to elevation, offering insights into potential climate change feedbacks.
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Ecossistema , Solo , Solo/química , Carbono , Florestas , Biomassa , MineraisRESUMO
Recently, with more portable diagnostic devices being moved to people anywhere, point-of-care (PoC) imaging has become more convenient and more popular than the traditional "bed imaging". Instant image segmentation, as an important technology of computer vision, is receiving more and more attention in PoC diagnosis. However, the image distortion caused by image preprocessing and the low resolution of medical images extracted by PoC devices are urgent problems that need to be solved. Moreover, more efficient feature representation is necessary in the design of instant image segmentation. In this paper, a new feature representation considering the relationships among local features with minimal parameters and a lower computational complexity is proposed. Since a feature window sliding along a diagonal can capture more pluralistic features, a Diagonal-Axial Multi-Layer Perceptron is designed to obtain the global correlation among local features for a more comprehensive feature representation. Additionally, a new multi-scale feature fusion is proposed to integrate nonlinear features with linear ones to obtain a more precise feature representation. Richer features are figured out. In order to improve the generalization of the models, a dynamic residual spatial pyramid pooling based on various receptive fields is constructed according to different sizes of images, which alleviates the influence of image distortion. The experimental results show that the proposed strategy has better performance on instant image segmentation. Notably, it yields an average improvement of 1.31% in Dice than existing strategies on the BUSI, ISIC2018 and MoNuSeg datasets.
RESUMO
PURPOSE: Investigate the most appropriate mathematical formula to objectively express upper eyelid contour symmetry. METHODS: 62 eyes of 31 patients were included in the study. The upper eyelid contour symmetry of the patients was classified subjectively (independent of MRD1) as poor, acceptable, and good by three oculoplastic specialists (senior, expert, and junior surgeon). Bézier curves of the upper lid contour were drawn with ImageJ software (NIH, Bethesda, MA, USA). Using the algorithms created by Author SKC in Spyder (Python 3.7.9.), the symmetry of the Bézier curves of the left eyelids were obtained according to the y-axis, and the mid-pupils of both eyes were superimposed. The lower curve moved vertically to the equal height of the other curve to equalize MRD1's. R2 (Coefficient of determination), RMSE (Root-mean-square error), MSE (Mean squared error), POC (Percentage of co-efficiency), and MAE (Mean absolute error) were calculated. We evaluated the correlation between these objective formulas and the subjective grading of three surgeons using Spearman's rho (ρ). RESULTS: The correlation coefficient of RMSE and MSE were the same for all surgeons grading. There was a strong correlation between the senior surgeon's subjective scoring (N; poor = 8, acceptable = 16, good = 8) and R2, RMSE, POC, MAE (ρ = 0.643, p < 0.001, ρ = -0.607, p < 0.001, ρ = 0.562, p < 0.001, ρ = -0.517, p < 0.001, respectively). We found a strong relationship between the expert surgeon's subjective scoring (N; poor = 9, acceptable = 13, good:10) and R2 (ρ = 0.611, p < 0.001), RMSE (ρ = -0.549, p < 0.001), POC (ρ = 0.511, p < 0.001), and MAE (ρ = -0.450, p < 0.05). We found a strong correlation between junior surgeon's subjective scoring (N; poor = 6, acceptable = 18, good = 8) and R2, RMSE, and POC (ρ: -0.517, p < 0.001; ρ: -0.470, p < 0.001; ρ: 0.521, p < 0.001; respectively) and moderate correlation between MAE (ρ:-0.394, p < 0.05). The highest correlation is observed with R2. CONCLUSIONS: RMSE, MSE, POC, MAE, and especially R2, may quantitatively express upper eyelid contour symmetry, comparable with the oculoplastic surgeon. The highest correlation was observed between the senior surgeon and R2, and decreases with the experience of the surgeon.