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1.
Discov Nano ; 19(1): 94, 2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38806816

RESUMO

In this research, we introduce an advanced methodology for the calculation of bulk light sources tailored for free-form surface design, focusing on the principle of energy conservation. This method is especially relevant for the evolving needs of micro-LED packaging, highlighting its potential in this burgeoning field. Our work includes the development of an algorithm for creating freeform-designed chip-scale package (FDCSP) components. These components seamlessly integrate LEDs and lenses, underscoring our commitment to advancing free-form surface design in chip-level packaging. By adhering to the principle of energy conservation, our approach facilitates a meticulous comparison of simulation outcomes with predefined target functions. This enables the iterative correction of discrepancies, employing layering techniques to refine the design until the simulated results closely align with our goals, as demonstrated by an appropriate difference curve. The practical application of these simulations leads to the innovative design of FDCSP devices. Notably, these devices are not just suitable for traditional applications in backlight modules but are explicitly optimized for the emerging sector of micro-LED packaging. Our successful demonstration of these FDCSP devices within backlight modules represents a significant achievement. It underscores the effectiveness of our design strategy and its expansive potential to transform micro-LED packaging solutions. This research not only contributes to the theoretical understanding of energy conservation in lighting design but also paves the way for groundbreaking applications in micro-LED and backlight module technologies.

2.
Anal Chem ; 96(8): 3578-3586, 2024 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-38354049

RESUMO

Existing mass spectrometric assays used for sensitive and specific measurements of target proteins across multiple samples, such as selected/multiple reaction monitoring (SRM/MRM) or parallel reaction monitoring (PRM), are peptide-based methods for bottom-up proteomics. Here, we describe an approach based on the principle of PRM for the measurement of intact proteoforms by targeted top-down proteomics, termed proteoform reaction monitoring (PfRM). We explore the ability of our method to circumvent traditional limitations of top-down proteomics, such as sensitivity and reproducibility. We also introduce a new software program, Proteoform Finder (part of ProSight Native), specifically designed for the easy analysis of PfRM data. PfRM was initially benchmarked by quantifying three standard proteins. The linearity of the assay was shown over almost 3 orders of magnitude in the femtomole range, with limits of detection and quantification in the low femtomolar range. We later applied our multiplexed PfRM assay to complex samples to quantify biomarker candidates in peripheral blood mononuclear cells (PBMCs) from liver-transplanted patients, suggesting their possible translational applications. These results demonstrate that PfRM has the potential to contribute to the accurate quantification of protein biomarkers for diagnostic purposes and to improve our understanding of disease etiology at the proteoform level.


Assuntos
Leucócitos Mononucleares , Proteínas , Humanos , Leucócitos Mononucleares/química , Reprodutibilidade dos Testes , Espectrometria de Massas , Proteômica/métodos , Processamento de Proteína Pós-Traducional , Proteoma/análise
3.
J Dairy Sci ; 107(6): 3738-3752, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38246544

RESUMO

In this study, we aimed to improve current udder health genetic evaluations by addressing the limitations of monthly sampled somatic cell score (SCS) for distinguishing cows with robust innate immunity from those susceptible to chronic infections. The objectives were to (1) establish novel somatic cell traits by integrating SCS and the differential somatic cell count (DSCC), which represents the combined proportion of polymorphonuclear leukocytes and lymphocytes in somatic cells and (2) estimate genetic parameters for the new traits, including their daily heritability and genetic correlations with milk production traits and SCS, using a random regression test-day model (RRTDM). We derived 3 traits, termed ML_SCS_DSCC, SCS_4_DSCC_65_binary, and ML_SCS_DSCC_binary, by using milk loss (ML) estimates at corresponding SCS and DSCC levels, thresholds established in previous studies, and a threshold established from milk loss estimates, respectively. Data consisted of test-day records collected during January 2021 through March 2022 from 265 herds in Hokkaido, Japan. From these records, we extracted records between 7 to 305 d in milk (DIM) in the first lactation to fit the RRTDM. The model included the random effect of herd-test-day, the fixed effect of year-month, fixed lactation curves nested with calving age groups, and random regressions with Legendre polynomials of order 3 for additive genetic and permanent environmental effects. The analysis was performed using Gibbs sampling with Gibbsf90+ software. The averages (ranges) of the daily heritability estimates over lactation were 0.086 (0.075-0.095) for SCS, 0.104 (0.073-0.127) for ML_SCS_DSCC, 0.137 (0.014-0.297) for SCS_4_DSCC_65_binary, and 0.138 (0.115-0.185) for ML_SCS_DSCC_binary; the heritability curve for SCS_4_DSCC_65_binary was erratic. Genetic correlations within the trait decreased as the DIM interval widened, especially for those integrating DSCC, indicating that these traits should be analyzed using RRTDM rather than repeatability models. The averages (ranges) of genetic correlations with milk yield over lactation were 0.01 (-0.22 to 0.28) for SCS, -0.05 (-0.40 to 0.13) for ML_SCS_DSCC, -0.08 (-0.17 to 0.09) for SCS_4_DSCC_65_binary, and -0.08 (-0.22 to 0.27) for ML_SCS_DSCC_binary. Compared with SCS, the newly defined traits exhibited slightly stronger negative genetic correlations with milk yield. Especially in late lactation stages, the genetic correlation between ML_SCS_DSCC and milk yield was significantly below zero, with a posterior median of -0.40. Furthermore, the new traits showed positive correlations with SCS, having estimates varying from 0.68 to 0.85 for ML_SCS_DSCC, 0.14 to 0.47 for SCS_4_DSCC_65_binary, and 0.61 to 0.66 for ML_SCS_DSCC_binary, depending on DIM. Considering that ML_SCS_DSCC and ML_SCS_DSCC_binary have relatively high heritability (compared with SCS) and favorable genetic correlations with milk production traits and SCS, their incorporation into breeding programs appears promising. Nevertheless, their genetic relationships with (sub)clinical mastitis require further investigation.


Assuntos
Lactação , Mastite Bovina , Leite , Animais , Lactação/genética , Feminino , Leite/citologia , Bovinos/genética , Mastite Bovina/genética , Contagem de Células/veterinária , Fenótipo , Japão , População do Leste Asiático
4.
J Vet Med Sci ; 86(1): 7-17, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37981317

RESUMO

Immune responses in bovine clinical mastitis (CM) probably differ depending on the causative pathogen and disease severity. The observational study aimed to investigate whether both factors are associated with the dynamics of immune indicators, including somatic cell score (SCS), white blood cell count (WBC), serum albumin/globulin (A/G) ratio, and differential somatic cell count (DSCC). We collected blood and milk samples 0, 3, 5, 7, 14, and 21 days after CM occurred in 38 cows, and grouped the cases (n=49) by disease severity and pathogen. We analyzed data using a linear mixed model considering the effects of pathogens and severity, calculated estimated-marginal means for indicators at each time point, and compared the means between groups. The dynamics of WBC varied depending on both pathogen and severity. WBC changed drastically in either severe or coliform-caused CM, slightly elevated in streptococcal mastitis, but unchanged in staphylococcal mastitis. This possibly relates to the deficiency in innate immune response toward staphylococci. The A/G ratio also changed depending on severity, as it dropped sharply only in severe CM. We observed a non-linear relationship between DSCC and SCS, possibly due to mammary epithelial cells shedding in milk when CM occurred. When cows recovering from Streptococcus dysgalatiae mastitis, DSCC decreased while SCS remained high, suggesting a healing process requiring more macrophages. Our results demonstrate that both the severity and pathogen are associated with immune responses in CM, providing insights into mastitis pathogenesis.


Assuntos
Doenças dos Bovinos , Mastite Bovina , Infecções Estreptocócicas , Feminino , Bovinos , Animais , Leite , Infecções Estreptocócicas/veterinária , Contagem de Leucócitos/veterinária , Imunidade , Contagem de Células/veterinária , Glândulas Mamárias Animais/patologia
5.
J Dairy Sci ; 106(11): 7942-7953, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37562643

RESUMO

This observational study aimed to use somatic cell score (SCS) and differential somatic cell count (DSCC), the combined proportion of polymorphonuclear leukocytes and lymphocytes in somatic cells, to investigate how mastitis affected milk production. Using generalized additive models, we analyzed 50,618 test-day records from 8,081 lactations from 7,912 cows in 197 herds between January 2021 and March 2022 to estimate the nonlinear interaction between SCS and DSCC, and the effects of lactation stages and seasons on milk yield, milk component percentages, and milk component yields by parity of cows. The results show that the interaction between SCS and DSCC on these traits was significant, nonlinear, and complex. When DSCC was high, the negative effects of SCS were minimal, even when SCS reached 8 (i.e., 3,200,000 somatic cells/mL). Cows with high DSCC could have milk yields similar to healthy cows, implying that these cows may have been in the early stages of mastitis and that the milk yield had yet to be affected. Contrastingly, when DSCC was low, milk loss due to high SCS was drastic, especially for cows in third or later lactations, whose milk yield could reduce from more than 35 kg/d to less than 15 kg/d (-59.9%). This tremendous milk loss in high-parity cows was likely due to their higher milk yield and higher risks of chronic mastitis. High SCS and low DSCC also led to a pronounced change in milk composition. The decrease in the percentage of lactose can be directly related to the damage of inflammation to the mammary gland, while the increase in fat and protein percentages was more attributable to the concentration effect resulting from the reduced milk yield. Compared with analyses based on categorized SCS and DSCC values, modeling these 2 indices directly helps us more precisely assess mastitis effects on milk yield and milk composition. For efficient milk production, our results indicate that we should prevent high-parity cows from entering a state of high SCS and low DSCC.

6.
J Cancer Res Clin Oncol ; 149(12): 10561-10583, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37286734

RESUMO

CLIC5 encoded protein associates with actin-based cytoskeletal and is increasingly thought to play significant roles in human cancers. We use TCGA and GEO to explore CLIC5 expression differences, mutation and DNA methylation, TMB, MSI, and immune cell infiltration. We verified the mRNA expression of CLIC5 in human ovarian cancer cells by real-time PCR and detected the expression of CLIC5 as well as immune marker genes in ovarian cancer by immunohistochemistry. The pan-cancer analysis showed that CLIC5 is highly expressed in several malignant tumors. In some cancers, CLIC5 expression in tumor samples is associated with poorer overall survival. For example, patients with ovarian cancer with high expression of CLIC5 have a poor prognosis. CLIC5 mutation frequency increased in all tumor types. The CLIC5 promoter is hypomethylated in most tumors. CLIC5 was associated with tumor immunity and different immune cells of different tumor types, such as CD8 + T cells, tumor-associated fibroblasts, macrophages, etc. CLIC5 was positively correlated with various immune checkpoints, and TMB and MSI were correlated with dysregulation of CLIC5 in tumors. The expression of CLIC5 in ovarian cancer was detected by qPCR and IHC, and the results were consistent with the bioinformatics results. There were a strong positive correlation between CLIC5 expression and M2 macrophage (CD163) infiltration and a negative correlation with CD8 + T-cell infiltration. In conclusions, our first pan-cancer analysis offered a detailed grasp of the cancerogenic functions of CLIC5 in a variety of malignancies. CLIC5 participated in immunomodulation and performed a crucial function in the tumor microenvironment.


Assuntos
Canais de Cloreto , Neoplasias Ovarianas , Feminino , Humanos , Fibroblastos Associados a Câncer , Linfócitos T CD8-Positivos , Canais de Cloreto/genética , Proteínas dos Microfilamentos , Neoplasias Ovarianas/genética , Prognóstico , Microambiente Tumoral
7.
JDS Commun ; 3(6): 441-445, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36465503

RESUMO

The California Mastitis Test (CMT) has been used to estimate total somatic cell count (SCC) levels in milk; however, milk with similar SCC levels occasionally shows inconsistent CMT results, which limits the use of the CMT. This observational study aimed to investigate how differential cell counts in milk influence the CMT in Holstein cattle through the novel parameters differential somatic cell count (DSCC) and macrophage proportion (MAC). We performed the CMT on d 0, 3, 5, 7, 14, and 21 after identifying mastitis, and simultaneously measured SCC, DSCC, and MAC at the quarter level. We followed 58 mastitis events occurring in 41 cows and obtained 307 quarter-level records after data cleaning. We transformed SCC to somatic cell score (SCS) and MAC to its logarithm to fit the normal distribution and analyzed the data using the cumulative logit mixed model. Results showed that both an increase in SCS (odds ratio: 3.66, 95% confidence interval: 2.89-4.64) and the logarithm of MAC (odds ratio: 4.35, 95% confidence interval: 1.91-9.91) can contribute to a higher CMT score. During the healing process of mastitis, MAC tends to increase as SCC decreases; thus, even samples with low SCC can cause positive CMT reactions. We recommend that practitioners avoid making treatment decisions based on the CMT alone. We also noted that the CMT is sensitive to subclinical mastitis with high MAC, hence it could be considered an alternative to detecting high MAC (chronic) mastitis.

8.
Sci Rep ; 12(1): 15095, 2022 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-36064958

RESUMO

Phosphorylation controls important cellular signals and its dysregulation leads to disease. While most phospho-regulation studies are focused on kinases, phosphatases are comparatively overlooked. Combining peptide arrays with SAMDI mass spectrometry, we show that tyrosine phosphatase activity is restricted by basic amino acids adjacent to phosphotyrosines. We validate this model using two ß-catenin mutants associated with cancer (T653R/K) and a mouse model for intellectual disability (T653K). These mutants introduce a basic residue next to Y654, an established phosphorylation site where modification shifts ß-catenin from cell-cell adhesions and towards its essential nuclear role as Wnt-signaling effector. We show that T653-basic mutant ß-catenins are less efficiently dephosphorylated by phosphatases, leading to sustained Y654 phosphorylation and elevated Wnt signals, similar to those observed for Y654E phospho-mimic mutant mice. This model rationalizes how basic mutations proximal to phosphotyrosines can restrict counter-regulation by phosphatases, providing new mechanismistic and treatment insights for 6000+ potentially relevant cancer mutations.


Assuntos
Proteínas Tirosina Fosfatases , beta Catenina , Animais , Camundongos , Mutação , Fosforilação , Proteínas Tirosina Fosfatases/genética , Proteínas Tirosina Fosfatases/metabolismo , Tirosina/metabolismo , Via de Sinalização Wnt , beta Catenina/genética , beta Catenina/metabolismo
9.
Anim Sci J ; 93(1): e13718, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35417050

RESUMO

Selective dry cow therapy (SDCT) allocates antibiotics in the dry period to cows or quarters with a high risk of intramammary infection (IMI), potentially improving antibiotic stewardship. However, SDCT has not been used in Japan, possibly due to concerns of negative impacts on udder health. This research aims to evaluate how the SDCT use affected dry period IMI dynamics in Japan. Additionally, the effects of management factors were also considered. At dry-off, 44 cows received antibiotics plus external teat sealant or external sealant in isolation based on their IMI risk, which was assessed using milk culture, a modified California mastitis test (CMT), and mastitis records. The SDCT approach allowed antibiotic use to be reduced by 33.7%. However, quarters with a low risk of infection who received no antibiotics had a numerically higher prevalence of IMI before calving than those who received antibiotics (28.6% vs. 19.2%). In addition, an increased risk of IMI in quarters without antibiotics was also attributed to poor herd hygiene and a shorter duration of external teat sealant adherence. The result suggests that these factors influence the outcome of an SDCT program. Therefore, a uniform recommendation for dry cow herd management may not be optimal.


Assuntos
Doenças dos Bovinos , Mastite Bovina , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Contagem de Células/veterinária , Feminino , Japão/epidemiologia , Lactação , Glândulas Mamárias Animais , Mastite Bovina/epidemiologia , Mastite Bovina/prevenção & controle , Leite
10.
Aging Cell ; 20(12): e13509, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34725922

RESUMO

Observational epidemiological studies have associated vitamin D deficiency with Alzheimer's disease (AD). However, whether vitamin D deficiency would result in some impacts on the vitamin D binding receptor (VDR) remains to be characterized in AD. Vitamin D helps maintain adult brain health genomically through binding with and activating a VDR/retinoid X receptor (RXR) transcriptional complex. Thus, we investigated the role of VDR in AD using postmortem human brains, APP/PS1 mice, and cell cultures. Intriguingly, although vitamin D was decreased in AD patients and mice, hippocampal VDR levels were inversely increased. The abnormally increased levels of VDR were found to be colocalized with Aß plaques, gliosis and autophagosomes, implicating a non-genomic activation of VDR in AD pathogenesis. Mechanistic investigation revealed that Aß upregulated VDR without its canonical ligand vitamin D and switched its heterodimer binding-partner from RXR to p53. The VDR/p53 complex localized mostly in the cytosol, increased neuronal autophagy and apoptosis. Chemically inhibiting p53 switched VDR back to RXR, reversing amyloidosis and cognitive impairment in AD mice. These results suggest a non-genomic rewiring of VDR to p53 is key for the progression of AD, and thus VDR/p53 pathway might be targeted to treat people with AD.


Assuntos
Doença de Alzheimer/genética , Doença de Alzheimer/fisiopatologia , Autofagia/genética , Proteína Supressora de Tumor p53/metabolismo , Deficiência de Vitamina D/complicações , Vitamina D/metabolismo , Animais , Humanos , Camundongos
11.
Sci Rep ; 11(1): 14491, 2021 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-34262095

RESUMO

We have studied carefully the behaviors of entangled qubits on the IBM Rochester with various connectivities and under a "noisy" environment. A phase trajectory analysis based on our measurements of the GHZ-like states is performed. Our results point to an important fact that entangled qubits are "protected" against environmental noise by a scaling property that impacts only the weighting of their amplitudes. The reproducibility of most measurements has been confirmed within a reasonably short gate operation time. But there still are a few combinations of qubits that show significant entanglement evolution in the form of transitions between quantum states. The phase trajectory of an entangled evolution, and the impact of the sudden death of GHZ-like states and the revival of newly excited states are analyzed in details. All observed trajectories of entangled qubits arise under the influences of the newly excited states in a "noisy" intermediate-scale quantum (NISQ) computer.

12.
J Chin Med Assoc ; 83(12): 1079-1086, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33009208

RESUMO

BACKGROUND: Sialic acids (SIAs), for example, α2,6-SIAs, can link to conserved N-glycans of immunoglobulin G (IgG). In this study, we investigated the correlation between α2,6-SIA on IgG and IgM and the disease activity of arthritis and rheumatoid arthritis (RA) in mice. METHODS: We measured α2,6-SIA levels in IgGs and IgMs in collagen-induced arthritis (CIA). Additionally, α2,6-SIA levels in rheumatoid factors (RFs) and anti-cyclic citrullinated peptide (anti-CCP) antibodies in RA patients were measured. Correlations between α2,6-SIA on Igs and CIA were analyzed and also in RA patients by utilizing the disease activity score 28 (DAS28). The ability to differentiate RA progression by Ig and autoantibody α2,6-SIA levels was examined. RESULTS: In CIA mice, plasma IgG-α2,6-SIA/IgG ratios decreased, whereas plasma IgM-α2,6-SIA/IgM ratios increased. Moreover, arthritis was not observed in collagen-injected mice with decreased IgG-α2,6-SIA/IgG ratios and without increased IgM-α2,6-SIA/IgM ratios. Isolated IgG-α2,6-SIA/IgG ratios displayed a significant inverse correlation with DAS28 scores (r = -0.383, p = 0.037). In contrast, isolated IgM-α2,6-SIA/IgM ratios correlated positively with DAS28 (r = 0.351, p = 0.009). Isolated IgG-anti-CCP-α2,6-SIA/plasma IgG-anti-CCP ratios were differentiated into either the remission (higher ratios) or the nonremission (lower ratios) category (p = 0.061), which is similar to the pattern for C-reactive protein (CRP) (p = 0.041) but different from that for the erythrocyte sedimentation rate (ESR) (p = 0.421). Using multiple linear regression analysis, plasma IgMRF-α2,6-SIA/IgMRF ratios displayed a correlation with DAS28 (p = 0.006), which was also observed in the ESR (p = 0.005), but was different from that for CRP (p = 0.222). CONCLUSION: Concurrent reverse expression of α2,6-SIA ratios on IgM and IgG correlated with the occurrence of CIA and RA disease activity. Thus, α2,6-SIA ratios on IgG-anti-CCP antibodies and IgMRF are potential markers for evaluating RA disease activities.


Assuntos
Artrite Reumatoide/sangue , Autoanticorpos/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Ácido N-Acetilneuramínico/análise , Adulto , Idoso , Animais , Anticorpos Antiproteína Citrulinada/sangue , Biomarcadores/sangue , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade
13.
ACS Comb Sci ; 21(11): 760-769, 2019 11 11.
Artigo em Inglês | MEDLINE | ID: mdl-31553163

RESUMO

The opposing activities of phosphatases and kinases determine the phosphorylation status of proteins, yet kinases have received disproportionate attention in studies of cellular processes, with the roles of phosphatases remaining less understood. This Research Article describes the use of phosphotyrosine-containing peptide arrays together with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to directly profile phosphatase substrate selectivities. Twenty-two protein tyrosine phosphatases were characterized with the arrays to give a profile of their specificities. An analysis of the data revealed that certain residues in the substrates had a conserved effect on activity for all enzymes tested, including the general rule that inclusion of a basic lysine or arginine residue on either side of the phosphotyrosine decreased activity. This insight also provides a new perspective on the role of a R1152Q mutant in the insulin receptor, which is known to exhibit a lower phosphorylation level and which this work suggests may be due to an increased activity toward phosphatase enzymes. The use of self-assembled monolayers for matrix-assisted laser desorption/ionization mass spectrometry (SAMDI-MS) to provide a rapid and quantitative assay of phosphatase enzymes will be important to gaining a more complete understanding of the biochemistry and biology of this important enzyme class.


Assuntos
Proteínas Tirosina Fosfatases/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Sequência de Aminoácidos , Peptídeos , Análise Serial de Proteínas , Manejo de Espécimes , Especificidade por Substrato
15.
Methods Enzymol ; 607: 389-403, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30149867

RESUMO

Phosphatases, the enzymes responsible for dephosphorylating proteins, play critical roles in many cellular processes. While their importance is widely recognized, phosphatase activity and regulation remain poorly understood. Currently, there are few assays available that are capable of directly measuring phosphatase activity and specificity. We have previously introduced SAMDI (self-assembled monolayers on gold for matrix-assisted laser desorption/ionization) mass spectrometry as a technique to profile the substrate specificities of enzymes. SAMDI mass spectrometry assays are well suited to examine phosphatase activities and offer many advantages over current methods. This technique uses monolayers that terminate with a peptide or molecular enzyme substrate and allows for enzyme reactions to be performed on a surface that can easily be rinsed and analyzed by mass spectrometry without the need for analyte labeling. In this chapter, we describe the process of combining SAMDI mass spectrometry with peptide arrays to study the substrate specificities of two protein tyrosine phosphatases.


Assuntos
Ensaios Enzimáticos/métodos , Monoéster Fosfórico Hidrolases/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Ensaios Enzimáticos/instrumentação , Ouro/química , Ensaios de Triagem em Larga Escala/instrumentação , Ensaios de Triagem em Larga Escala/métodos , Peptídeos/análise , Peptídeos/química , Peptídeos/metabolismo , Monoéster Fosfórico Hidrolases/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentação , Especificidade por Substrato
16.
Nanoscale ; 10(13): 6214, 2018 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-29595205

RESUMO

Correction for 'A high quality liquid-type quantum dot white light-emitting diode' by Chin-Wei Sher et al., Nanoscale, 2016, 8, 1117-1122.

17.
Angew Chem Int Ed Engl ; 56(52): 16531-16535, 2017 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-28940795

RESUMO

Identifying peptide substrates that are efficiently cleaved by proteases gives insights into substrate recognition and specificity, guides development of inhibitors, and improves assay sensitivity. Peptide arrays and SAMDI mass spectrometry were used to identify a tetrapeptide substrate exhibiting high activity for the bacterial outer-membrane protease (OmpT). Analysis of protease activity for the preferred residues at the cleavage site (P1, P1') and nearest-neighbor positions (P2, P2') and their positional interdependence revealed FRRV as the optimal peptide with the highest OmpT activity. Substituting FRRV into a fragment of LL37, a natural substrate of OmpT, led to a greater than 400-fold improvement in OmpT catalytic efficiency, with a kcat /Km value of 6.1×106  L mol-1 s-1 . Wild-type and mutant OmpT displayed significant differences in their substrate specificities, demonstrating that even modest mutants may not be suitable substitutes for the native enzyme.

18.
Cell Physiol Biochem ; 42(4): 1701-1711, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28743112

RESUMO

BACKGROUND/AIMS: MiR-302b is a major microRNA found in human embryonic stem cells and induced pluripotent stem cells. However, its function in gastric cancer progression remains unclear. METHODS: Quantitative reverse transcription-PCR was performed to detect the expression levels of miR-302b-3p in gastric cancer tissues. MTT, colony formation, and flow cytometer analyses were conducted to explore the function of miR-302b-3p in MKN-45/SGC-7901 cells. A dual-luciferase reporter was used to validate the bioinformatics-predicted target gene of miR-302b-3p. Western blotting and RNA interference were used to evaluate the expression of the AKT signaling pathway and determine the mechanisms underlying miR-302b-3p-induced anti-tumor effects. RESULTS: MiR-302b-3p expression was decreased in gastric cancer tissues and cell lines. Enforced expression of miR-302b suppressed cell proliferation and cell cycle G1-S transition and induced apoptosis. IGF-1R was found to be a direct target of miR-302b-3p, and silencing of IGF-1R resulted in the same biological effects as those induced by miR-302b-3p overexpression in gastric cancer cells. Importantly, both overexpression of miR-302b-3p and silencing of IGF-1R decreased AKT phosphorylation, which modulated AKT related cell cycle regulators (cyclin A2, cyclin D1, CDK2, and CDk6) and apoptotic protein Bax/Bcl-2. CONCLUSION: These results indicate the tumor suppressor role of miR-302b-3p in the pathogenesis of gastric cancer.


Assuntos
Adenocarcinoma/genética , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Proteínas Proto-Oncogênicas c-akt/genética , Receptor IGF Tipo 1/genética , Neoplasias Gástricas/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Apoptose/genética , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células , Ciclina A2/genética , Ciclina A2/metabolismo , Ciclina D1/genética , Ciclina D1/metabolismo , Quinase 2 Dependente de Ciclina/genética , Quinase 2 Dependente de Ciclina/metabolismo , Quinase 6 Dependente de Ciclina/genética , Quinase 6 Dependente de Ciclina/metabolismo , Genes Reporter , Humanos , Luciferases/genética , Luciferases/metabolismo , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Receptor IGF Tipo 1/antagonistas & inibidores , Receptor IGF Tipo 1/metabolismo , Transdução de Sinais , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
19.
Altern Ther Health Med ; 22(6): 20-23, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27866177

RESUMO

Background • Highly concentrated cranberry juice has long been considered to have protective properties against urinary tract infections (UTIs), on the basis of its content of cranberry proanthocyanidins, with A-type interflavan bonds. Objective • This study intended to evaluate the benefits of a highly concentrated cranberry juice for the prevention of repeated episodes of UTI in uncircumcised boys. Design • The study was a randomized, controlled trial. Setting • The study took place at Taipei City Hospital, Renai and Zhongxing Branches (Taipei City, Taiwan). Participants • Participants were 55 uncircumcised boys and 12 circumcised boys, aged 6 to 18 y, with histories of uncomplicated UTI, who were patients at the hospital. Intervention • The uncircumcised boys were randomly divided into 2 groups: (1) group 1 (n = 28) took 4 oz (120 mL) daily of cranberry juice for 6 mo; and (2) group 2 (n = 27), the negative control group, drank a placebo juice for 6 months. The circumcised boys in group 3, a positive control group, also drank a placebo juice for 6 mo. Outcome Measures • The time to UTI (ie, to the appearance of symptoms plus pyuria) was the main outcome. Asymptomatic bacteriuria, adherence to the treatment, and adverse effects were assessed at monthly visits. Results • After 6 mo of a prophylactic treatment with cranberry juice, the incidence of bacteriuria, mainly Escherichia coli, as shown in urine cultures at ≥1 × 105, were 25% (7/28), 37% (10/27), and 33.3% (4/12) in groups 1, 2 and 3, respectively. The comparisons of the rate of prevention of a recurrence of UTI between group 1 and group 2 and between group 1 and group 3 showed that group 1 had fewer recurrent episodes of UTI. No children withdrew from the study. No adverse events or side effects were recorded. Conclusions • Cranberry juice may reduce the number of repeated episodes of UTI in uncircumcised boys and may have beneficial effects against the growth of Gram-negative bacterial pathogens. Its preventive benefits against UTI in the uncircumcised boys were even higher than those of circumcision for the circumcised boys.


Assuntos
Circuncisão Masculina , Sucos de Frutas e Vegetais , Infecções Urinárias/prevenção & controle , Vaccinium macrocarpon , Adolescente , Criança , Humanos , Masculino , Recidiva , Resultado do Tratamento
20.
PLoS One ; 11(3): e0151669, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26981635

RESUMO

OBJECTIVE: We attempted to determine whether the level of enzymes sialyltransferase (ST) and neuraminidase (Neu) and sialic acid (SIA) in patients with systemic lupus erythematosus (SLE) correlates with the SLE Disease Activity Index (SLEDAI) and in patients with rheumatoid arthritis (RA) correlates with the Disease Activity Score28 (DAS28). METHODS: We examined cell-surface levels of ST6Gal-1, Neu1, ST3Gal-1, Neu3, α-2,6-SIA, and α-2,3-SIA by using fluorescent anti-enzyme antibodies, fluorescent-conjugated Sambucus nigra lectin, and fluorescent-conjugated Maackia amurensis lectin on blood cells in SLE and RA patients and assessed correlations of these levels with SLEDAI and with DAS28. Areas under the curve (AUC) were calculated for different variables against SLEDAI. RESULTS: The B-cell ST3Gal-1/Neu3 ratio positively correlated with SLEDAI scores (ρ = 0.409 and P = 0.002, statistically significant after Bonferroni' correction for multiple analyses.). It was supported by the inverse correlation of B-cell Neu3 levels with SLEDAI scores (ρ = -0.264, P = 0.048). The B-cell ST3Gal-1/Neu3 ratio against SLEDAI yielded an AUC of 0.689, which was comparable to that of anti-dsDNA levels at 0.635. In contrast, both ST3Gal-1 and Neu3 levels of RA B cells (r = 0.376, P = 0.013; r = 0.425, P = 0.005, respectively) correlated positively with high disease-activity DAS28 scores. CONCLUSION: B-cell ST3Gal-1/Neu3 ratios in SLE and B-cell ST3Gal-1 and Neu3 levels in RA with high disease-activity DAS28 scores correlated with disease activity measures and may be useful in monitoring disease activities.


Assuntos
Artrite Reumatoide/diagnóstico , Linfócitos B/metabolismo , Lúpus Eritematoso Sistêmico/diagnóstico , Ácido N-Acetilneuramínico/metabolismo , Neuraminidase/sangue , Sialiltransferases/sangue , Adulto , Idoso , Artrite Reumatoide/sangue , Feminino , Humanos , Lúpus Eritematoso Sistêmico/sangue , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Índice de Gravidade de Doença , Adulto Jovem
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