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1.
Osteoporos Sarcopenia ; 10(1): 3-10, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38690538

RESUMO

Objectives: This study aimed to present the Asia-Pacific consensus on long-term and sequential therapy for osteoporosis, offering evidence-based recommendations for the effective management of this chronic condition. The primary focus is on achieving optimal fracture prevention through a comprehensive, individualized approach. Methods: A panel of experts convened to develop consensus statements by synthesizing the current literature and leveraging clinical expertise. The review encompassed long-term anti-osteoporosis medication goals, first-line treatments for individuals at very high fracture risk, and the strategic integration of anabolic and antiresorptive agents in sequential therapy approaches. Results: The panelists reached a consensus on 12 statements. Key recommendations included advocating for anabolic agents as the first-line treatment for individuals at very high fracture risk and transitioning to antiresorptive agents following the completion of anabolic therapy. Anabolic therapy remains an option for individuals experiencing new fractures or persistent high fracture risk despite antiresorptive treatment. In cases of inadequate response, the consensus recommended considering a switch to more potent medications. The consensus also addressed the management of medication-related complications, proposing alternatives instead of discontinuation of treatment. Conclusions: This consensus provides a comprehensive, cost-effective strategy for fracture prevention with an emphasis on shared decision-making and the incorporation of country-specific case management systems, such as fracture liaison services. It serves as a valuable guide for healthcare professionals in the Asia-Pacific region, contributing to the ongoing evolution of osteoporosis management.

2.
Osteoporos Sarcopenia ; 10(1): 22-27, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38690543

RESUMO

Objectives: Vertebral fracture is both common and serious among adults, yet it often goes undiagnosed. This study aimed to develop a shape-based algorithm (SBA) for the automatic identification of vertebral fractures. Methods: The study included 144 participants (50 individuals with a fracture and 94 without a fracture) whose plain thoracolumbar spine X-rays were taken. Clinical diagnosis of vertebral fracture (grade 0 to 3) was made by rheumatologists using Genant's semiquantitative method. The SBA algorithm was developed to determine the ratio of vertebral body height loss. Based on the ratio, SBA classifies a vertebra into 4 classes: 0 = normal, 1 = mild fracture, 2 = moderate fracture, 3 = severe fracture). The concordance between clinical diagnosis and SBA-based classification was assessed at both person and vertebra levels. Results: At the person level, the SBA achieved a sensitivity of 100% and specificity of 62% (95% CI, 51%-72%). At the vertebra level, the SBA achieved a sensitivity of 84% (95% CI, 72%-93%), and a specificity of 88% (95% CI, 85%-90%). On average, the SBA took 0.3 s to assess each X-ray. Conclusions: The SBA developed here is a fast and efficient tool that can be used to systematically screen for asymptomatic vertebral fractures and reduce the workload of healthcare professionals.

3.
Protein Sci ; 33(3): e4910, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38358125

RESUMO

Fluorinated aromatic amino acids (FAAs) are promising tools when studying protein structure and dynamics by NMR spectroscopy. The incorporation FAAs in mammalian expression systems has been introduced only recently. Here, we investigate the effects of FAAs incorporation in proteins expressed in human cells, focusing on the probability of incorporation and its consequences on the 19 F NMR spectra. By combining 19 F NMR, direct MS and x-ray crystallography, we demonstrate that the probability of FAA incorporation is only a function of the FAA concentration in the expression medium and is a pure stochastic phenomenon. In contrast with the MS data, the x-ray structures of carbonic anhydrase II reveal that while the 3D structure is not affected, certain positions lack fluorine, suggesting that crystallization selectively excludes protein molecules featuring subtle conformational modifications. This study offers a predictive model of the FAA incorporation efficiency and provides a framework for controlling protein fluorination in mammalian expression systems.


Assuntos
Aminoácidos , Proteínas , Animais , Humanos , Aminoácidos/química , Proteínas/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Flúor/química , Mamíferos
4.
Sci Transl Med ; 16(731): eadi3883, 2024 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-38266106

RESUMO

We previously described an approach called RealSeqS to evaluate aneuploidy in plasma cell-free DNA through the amplification of ~350,000 repeated elements with a single primer. We hypothesized that an unbiased evaluation of the large amount of sequencing data obtained with RealSeqS might reveal other differences between plasma samples from patients with and without cancer. This hypothesis was tested through the development of a machine learning approach called Alu Profile Learning Using Sequencing (A-PLUS) and its application to 7615 samples from 5178 individuals, 2073 with solid cancer and the remainder without cancer. Samples from patients with cancer and controls were prespecified into four cohorts used for model training, analyte integration, and threshold determination, validation, and reproducibility. A-PLUS alone provided a sensitivity of 40.5% across 11 different cancer types in the validation cohort, at a specificity of 98.5%. Combining A-PLUS with aneuploidy and eight common protein biomarkers detected 51% of the cancers at 98.9% specificity. We found that part of the power of A-PLUS could be ascribed to a single feature-the global reduction of AluS subfamily elements in the circulating DNA of patients with solid cancer. We confirmed this reduction through the analysis of another independent dataset obtained with a different approach (whole-genome sequencing). The evaluation of Alu elements may therefore have the potential to enhance the performance of several methods designed for the earlier detection of cancer.


Assuntos
Neoplasias , Humanos , Reprodutibilidade dos Testes , Neoplasias/diagnóstico , Neoplasias/genética , Elementos Nucleotídeos Curtos e Dispersos , Aprendizado de Máquina , Aneuploidia
5.
Environ Sci Technol ; 58(3): 1669-1679, 2024 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-38183301

RESUMO

Peatlands are known sinks for arsenic (As). In the present study, seasonal As mobilization was observed in an acidic, minerotrophic peatland (called Lehstenbach) in late summer, accompanied by a peak in dissolved sulfide (S(-II)). Arsenic speciation revealed the lowest seasonal porewater concentrations of arsenite and arsenate, likely due to As(III)-S-bridging to natural organic matter. Arsenic mobilization was driven by the formation of arsenite-S(-II) colloids and formation of methylthiolated arsenates (up to 59% of the sum of As species) and to a minor extent also of inorganic thioarsenates (6%-30%) and oxymethylated arsenates (5%-24%). Sorption experiments using a purified model peat, the Lehstenbach peat, natural (to mimic winter conditions) and reacted with S(-II) (to mimic late summer conditions) at acidic and neutral pH confirmed low sorption of methylthiolated arsenates. At acidic pH and in the presence of S(-II), oxymethylated arsenates were completely thiolated. This methylthiolation decreased As sorption up to 10 and 20 times compared with oxymethylated arsenates and arsenite, respectively. At neutral pH, thiolation of monomethylated arsenates was incomplete, and As could be partially retained as oxymethylated arsenates. Dimethylated arsenate was still fully thiolated and highly mobile. Misidentification of methylthiolated arsenates as oxymethylated arsenates might explain previous contradictory reports of methylation decreasing or increasing As mobility.


Assuntos
Arsênio , Arsenitos , Arseniatos , Estações do Ano , Solo
6.
Sci Rep ; 14(1): 32, 2024 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-38168502

RESUMO

The epidemiology of vertebral fractures (VF) in underrepresented populations is not well-documented. This cohort study was part of a longitudinal osteoporosis research project with the aim of determining the prevalence, incidence, and risk factors for VF. 401 individuals (155 men) aged 50 years and older without a clinical diagnosis of VF were took radiographs at baseline and 2 years later. VF were ascertained using the Genant's semi-quantitative method. Bone mineral density (BMD) of femoral neck and lumbar spine were measured by dual-energy X-ray absorptiometry (Hologic Inc). The association between VF and risk factors was analyzed by the multiple logistic regression. The 95% confidence interval for prevalence and incidence was estimated by exact Poisson test. At baseline, the prevalence of VF was 12.2% (n = 49, 95% CI 9.0-16.2%) and increased with advancing age with one-fifth of those aged 70 and older having a VF. During the follow-up period, we observed 6 new VF, making the incidence of 6.6/1000 person-years (n = 6, 95% CI 2.4-14.3). The risk of prevalent VF was associated with male gender (OR: 2.67; 95% CI 1.28-5.87) and T-score at the femoral neck (OR per one SD decrease: 1.1; 1.03-1.17). These data indicate that VF is common among adults, and that lower femoral neck BMD was a risk factor for VF.


Assuntos
Osteoporose , Fraturas da Coluna Vertebral , Adulto , Masculino , Humanos , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Fraturas da Coluna Vertebral/diagnóstico por imagem , Fraturas da Coluna Vertebral/epidemiologia , Fraturas da Coluna Vertebral/etiologia , Estudos de Coortes , Prevalência , Incidência , Vietnã , Osteoporose/diagnóstico por imagem , Osteoporose/epidemiologia , Osteoporose/complicações , Densidade Óssea , Absorciometria de Fóton/métodos , Fatores de Risco , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/lesões
7.
Sci Rep ; 14(1): 17, 2024 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-38168916

RESUMO

Sarcopenia, a condition characterized by muscle mass decline, is one of the leading health problems in the elderly. This study determined the rate of sarcopenia according to criteria by Asian Working Group for Sarcopenia (AWGS) and related factors in elderly people. A community-based cross-sectional study was conducted in 632 people aged 60 years or over in Ho Chi Minh City. Data were collected through a predefined questionnaire and direct measurement. Sarcopenia was identified based on the Inbody 770 machine and AWGS criteria. The prevalence of sarcopenia was 32.0%. Participants with advanced age, low education, unemployment, low level of family economics and frailty were more likely to have sarcopenia. Among these, frailty had the highest impact on sarcopenia, with significantly higher odds of having sarcopenia found in participants with pre-frailty (OR = 4.80, 95% CI 2.75-8.38, p < 0.001) and frailty (OR = 21.16, 95% CI 8.96-49.97, p < 0.001). In contrast, BMI was inversely associated with sarcopenia. Sarcopenia is prevalent in the Vietnamese elderly. Apart from social demographic characteristics including age, education, employment and family economic status, frailty appeared to be an important risk factor. Early screening, referral, and treatment of sarcopenia among the elderly having a high risk of sarcopenia are recommended.


Assuntos
Fragilidade , Sarcopenia , Idoso , Humanos , Sarcopenia/diagnóstico , Fragilidade/epidemiologia , Fragilidade/diagnóstico , Prevalência , Estudos Transversais , Vietnã/epidemiologia
8.
Macromol Biosci ; 24(2): e2300225, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37770246

RESUMO

In this study, stimuli-responsive liberation of an epidermal growth factor fragment (EGFfr) is accomplished using nanofibrous meshes to improve wound healing effects. Electrospun nanofibers are fragmented by mechanical milling, followed by aminolysis to fabricate powdered nanofibrils (NFs). EGFfrs are covalently immobilized on NFs via thioketal linkers (EGFfr@TK@NF) for reactive oxygen species (ROS)-dependent liberation. EGFfr@TK@NF exhibits ROS-responsive liberation of EGFfr from the matrix at hydrogen peroxide (H2 O2 ) concentrations of 0-250 mm. Released EGFfr is confirmed to enhance the migration of HaCaT cell monolayers, and keratinocytic gene expression levels are significantly enhanced when H2 O2 is added to obtain the released fraction of NFs. An in vivo study on the dorsal wounds of mice reveals that EGFfr-immobilized NFs improve the expression levels of keratin1, 5, and 14 for 2 weeks when H2 O2 is added to the wound sites, suggesting that the wounded skin is re-epithelized with the original epidermis. Thus, EGFfrs-immobilized NFs are anticipated to be potential nanotherapeutics for wound treatment in combination with the conventional disinfection process with H2 O2 .


Assuntos
Fator de Crescimento Epidérmico , Nanofibras , Camundongos , Animais , Fator de Crescimento Epidérmico/farmacologia , Espécies Reativas de Oxigênio , Cicatrização
9.
J Cachexia Sarcopenia Muscle ; 15(1): 380-386, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38146138

RESUMO

BACKGROUND: Sarcopenia is a geriatric disease characterized by the progressive and generalized loss of skeletal lean mass and strength with age. The prevalence of sarcopenia in the Vietnamese population is unknown. This study sought to estimate the prevalence of and risk factors for sarcopenia among community-dwelling individuals in Vietnam. METHODS: This cross-sectional study is part of the ongoing Vietnam Osteoporosis Study project. The study involved 1308 women and 591 men aged 50 years and older as at 2015 (study entry). Whole-body dual-energy X-ray absorptiometry was used to measure the appendicular skeletal lean mass. Anthropometric and clinical data were collected using a structured questionnaire. Sarcopenia was defined according to the criteria proposed by the Asian Working Group for Sarcopenia in 2019. Logistic regression analysis was used to determine the association between potential risk factors and sarcopenia. RESULTS: The prevalence of sarcopenia in women and men was 14% (n = 183) and 16% (n = 83), respectively. Age (odds ratio [OR] per 10 years = 1.37; 95% confidence interval [CI] 1.26-1.48) and being underweight (OR = 1.61; 95% CI 1.00-2.58) were independently associated with increased risk of sarcopenia. The combination of low physical activity, being underweight and advancing age accounted for ~27% of sarcopenic patients. However, most of the attributable fraction was due to ageing. CONCLUSIONS: Sarcopenia is common in community-dwelling Vietnamese adults, particularly those with advancing age, who are underweight and with low physical activity.


Assuntos
Osteoporose , Sarcopenia , Masculino , Adulto , Humanos , Feminino , Pessoa de Meia-Idade , Idoso , Criança , Sarcopenia/etiologia , Vietnã/epidemiologia , Prevalência , Vida Independente , Magreza/complicações , Estudos Transversais , Osteoporose/epidemiologia , Osteoporose/etiologia , Fatores de Risco
10.
Sci Rep ; 13(1): 18554, 2023 10 29.
Artigo em Inglês | MEDLINE | ID: mdl-37899423

RESUMO

High grade B-cell lymphoma with MYC and BCL2 rearrangements (HGBCL-DH) represents an uncommon B-cell lymphoma (BCL) with aggressive clinical courses and poor prognosis. Despite revolutionary therapeutic advances in BCL, there has been limited treatment progress in HGBCL-DH, thus necessitating additional therapeutic strategies for HGBCL-DH. This study demonstrated that the BET antagonist INCB057643 synergized with the XPO1 inhibitors (selinexor and eltanexor) to decrease cell viability and increase cell apoptosis in HGBCL-DH cells with or without TP53 mutations. As anticipated, the combined treatment of INCB057643 with selinexor slowed tumor growth and reduced the tumor burden in TP53-mutated HGBCL-DH xenografts. Mechanistically, MYC functional inhibition was a potential molecular mechanism underlying the synergy of the combined INCB057643 and selinexor treatment in HGBCL-DH cells independent of TP53 mutation status. In TP53 mutated HGBCL-DH cells, inducing DNA damage and impairing the DNA damage response (DDR) were involved in the therapeutic interaction of the combined regimen. In TP53 wild-type cells, the molecular mechanism was linked with upregulation of p53 levels and activation of its targeted pathways, rather than dysregulation of the DDR. Collectively, we might provide a potential promising combination therapy regimen for the management of HGBCL-DH. Clinical evaluations are warranted to confirm this conclusion.


Assuntos
Linfoma de Células B , Linfoma Difuso de Grandes Células B , Humanos , Regulação para Baixo , Linfoma de Células B/genética , Linfoma Difuso de Grandes Células B/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-myc/genética
11.
Anim Sci J ; 94(1): e13875, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37818840

RESUMO

The D-loop region on mitochondrial DNA (mtDNA) is frequently used for analyses of maternal lineages within domestic animal species. There are many native pig breeds in Vietnam, but their origins remain unclear. This study investigated maternal lineages using the D-loop region on mtDNA of 260 samples collected from native pigs in 20 provinces across Vietnam. The D-loop region of all samples was amplified and sequenced. We obtained 713 bp sequences of the D-loop region for each sample excluding the repeat region, and variants on this region were used to construct a phylogenetic tree. We detected 50 haplotypes from Vietnamese native pigs, with 27 novel haplotypes. Phylogenetic tree analysis showed two haplotype groups: one for the MTSEA group, frequently found in domestic pigs in the mountainous areas of Cambodia and Laos; and the D2 group, found in pigs originating from Chinese pigs. No European haplotype was found. Haplotypes in northeast Vietnam comprised only haplotypes of the D2 group, whereas in areas from the northwest mountains to the south, we found haplotypes belonging to both the D2 and MTSEA groups. This study suggested that both origins contributed to maternal lineages of current populations of Vietnamese native pigs.


Assuntos
DNA Mitocondrial , Variação Genética , Animais , DNA Mitocondrial/genética , Haplótipos/genética , Filogenia , Sus scrofa/genética , Suínos/genética , Vietnã
12.
J Air Waste Manag Assoc ; 73(8): 625-637, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37378540

RESUMO

Nowadays, when the zero-waste strategy is an inevitable component of the circular economy, the reuse of waste, including dredged sludges, has drawn the attention of many researchers. This study evaluated four kinds of bio-wastes (corn core powder, rice husk powder, sugarcane bagasse powder, and peanut shell powders) and two kinds of construction wastes (autoclaved aerated concrete-AAC and pavement stone) in enhancing the dewaterability of dredged sludge from the lake, in which the sludges would then be reused for brick production. The results showed that the moisture contents decreased from 62 ± 0.14% to 57 ± 1.89% after mixing and then to 35 ± 8.31% after compressing for the construction waste-blended sludge. Among the bio-wastes, the sugarcane bagasse additive performed the best at a mixing ratio of 1:3 by weight and rice husk powder worked best at a mixing ratio of 1:5 by weight. The organic matter was increased up to 80% when the bio-wastes were added, while it was decreased to 5% for the case of construction wastes. The optimum percentage of sludge in the mixture to meet all the oxide contents in the brick and energy saving shall be about 30%. The results have revealed a potentially green route for brick production with lake sediment and bio-waste/construction wastes.Implications: It is the first time the reuse of agro-wastes/construction waste was evaluated to mix with lake sediment to partly replace clay for brick production; Among the bio-wastes, the sugarcane bagasse additive performed the best at a mixing ratio of 1:3 by weight; Moisture contents decreased from 62 ± 0.14% to 57 ± 1.89% after mixing and then to 35 ± 8.31% after compressing for the blended sludge; The optimum percentage of mixed sludge, possibly replaced the clay in brick production, considering oxide contents and energy saving shall be up to 30%.


Assuntos
Saccharum , Esgotos , Argila , Celulose , Lagos , Pós , Materiais de Construção , Óxidos
13.
Genes (Basel) ; 14(5)2023 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-37239448

RESUMO

Indigenous chicken breeds have both cultural significance and economic value since they possess unique genetic characteristics that enable them to adapt to the local environment and contribute to biodiversity, food security, and sustainable agriculture in Vietnam. To (Tò in Vietnamese) chicken, a Vietnamese indigenous chicken breed, is popularly raised in Thai Binh province; however, little known is about the genetic diversity of this breed. In this study, we sequenced the complete mitochondrial genome of To chicken for a better understanding of the diversity and origin of the breed. The results of sequencing showed that the mitochondrial genome of To chicken spans a total length of 16,784 base pairs and comprises one non-coding control region (known as the displacement-loop (D-loop) region), two ribosomal RNA genes, 13 protein-coding genes, and 22 transfer RNA genes. The phylogenetic tree analyses and estimated genetic distances based on 31 complete mitochondrial genome sequences indicated that To chicken has a close genetic distance with the Laotian native chicken breed, Lv'erwu breed in China, and Nicobari black and Kadaknath breeds in India. The result of the current study might be important for conservation, breeding, and further genetic studies of To chicken.


Assuntos
Variação Genética , Genoma Mitocondrial , Animais , Filogenia , Galinhas/genética , Vietnã
14.
Proc Natl Acad Sci U S A ; 120(15): e2220704120, 2023 04 11.
Artigo em Inglês | MEDLINE | ID: mdl-37014860

RESUMO

The analysis of cell-free DNA (cfDNA) from plasma offers great promise for the earlier detection of cancer. At present, changes in DNA sequence, methylation, or copy number are the most sensitive ways to detect the presence of cancer. To further increase the sensitivity of such assays with limited amounts of sample, it would be useful to be able to evaluate the same template molecules for all these changes. Here, we report an approach, called MethylSaferSeqS, that achieves this goal, and can be applied to any standard library preparation method suitable for massively parallel sequencing. The innovative step was to copy both strands of each DNA-barcoded molecule with a primer that allows the subsequent separation of the original strands (retaining their 5-methylcytosine residues) from the copied strands (in which the 5-methylcytosine residues are replaced with unmodified cytosine residues). The epigenetic and genetic alterations present in the DNA molecules can then be obtained from the original and copied strands, respectively. We applied this approach to plasma from 265 individuals, including 198 with cancers of the pancreas, ovary, lung, and colon, and found the expected patterns of mutations, copy number alterations, and methylation. Furthermore, we could determine which original template DNA molecules were methylated and/or mutated. MethylSaferSeqS should be useful for addressing a variety of questions relating genetics and epigenetics.


Assuntos
Variações do Número de Cópias de DNA , Neoplasias , Feminino , Humanos , Metilação , 5-Metilcitosina , DNA/genética , Mutação , Neoplasias/genética , Metilação de DNA
15.
Osteoporos Int ; 34(8): 1381-1387, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37106043

RESUMO

Based on the prospective cohort of the Vietnam Osteoporosis Study, we show that bone loss occurred before menopause, and that the loss accelerated in the first 5 years post-menopause. PURPOSE: To define the change in bone mineral density (BMD) among women during the menopausal transition. METHODS: The study involved 1062 women aged 40-59 who were participants of the population-based prospective Vietnam Osteoporosis Study. BMD at the femoral neck (FN), lumbar spine (LS), and whole body scan was measured by DXA. Each woman has had two BMD measurements separated by approximately 2 years, and the rate of BMD change was calculated for each woman. Multivariable linear regression models were used to quantify the association between body composition parameters and the rate of BMD change. RESULTS: At FN, there were 3 phases of BMD change: a slight decline before the age of 45-49 (average loss of 0.51%/year); a substantial decline between the ages of 49 and 54 (average loss of 1.39%/year); and then slowed down between the ages of 54 and 59 (average loss of 0.31%/year). The same trend was also observed at LS: a slight decline (- 0.56%/year) among women aged 45-49; then a significant decline between the ages of 50 and 54 (- 1.33%/year); but then slowed down at - 0.31%/year after the age of 55. Changes in BMD were not significantly associated with changes in lean mass or fat mass. CONCLUSIONS: Although bone loss occurred before menopause, the loss accelerated in the early perimenopausal transition (45-50 years of age). This finding suggests that screening for osteoporosis in women should be considered at the age of 45.


Assuntos
Osteoporose Pós-Menopausa , Osteoporose , Feminino , Humanos , Pessoa de Meia-Idade , Densidade Óssea , Perimenopausa , Estudos Prospectivos , Vietnã/epidemiologia , Osteoporose/epidemiologia , Osteoporose/etiologia , Vértebras Lombares , Osteoporose Pós-Menopausa/epidemiologia , Colo do Fêmur
16.
J Am Chem Soc ; 145(2): 1389-1399, 2023 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-36604341

RESUMO

In-cell NMR spectroscopy is a powerful approach to study protein structure and function in the native cellular environment. It provides precious insights into the folding, maturation, interactions, and ligand binding of important pharmacological targets directly in human cells. However, its widespread application is hampered by the fact that soluble globular proteins often interact with large cellular components, causing severe line broadening in conventional heteronuclear NMR experiments. 19F NMR can overcome this issue, as fluorine atoms incorporated in proteins can be detected by simple background-free 1D NMR spectra. Here, we show that fluorinated amino acids can be easily incorporated in proteins expressed in human cells by employing a medium switch strategy. This straightforward approach allows the incorporation of different fluorinated amino acids in the protein of interest, reaching fluorination efficiencies up to 60%, as confirmed by mass spectrometry and X-ray crystallography. The versatility of the approach is shown by performing 19F in-cell NMR on several proteins, including those that would otherwise be invisible by 1H-15N in-cell NMR. We apply the approach to observe the interaction between an intracellular target, carbonic anhydrase 2, and its inhibitors, and to investigate how the formation of a complex between superoxide dismutase 1 and its chaperone CCS modulates the interaction of the chaperone subunit with the cellular environment.


Assuntos
Flúor , Chaperonas Moleculares , Humanos , Espectroscopia de Ressonância Magnética/métodos , Ressonância Magnética Nuclear Biomolecular/métodos , Flúor/química , Aminoácidos
17.
Am J Cancer Res ; 12(10): 4666-4679, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36381323

RESUMO

FOXM1 is a transcription factor that controls cell cycle regulation, cell proliferation, and differentiation. Overexpression of FOXM1 has been implicated in various cancer types. However, the activation status and functional significance of FOXM1 in diffuse large B cell lymphoma (DLBCL) have not been well investigated. Using proteomic approaches, we discovered that the protein expression levels of FOXM1 and PLK1 were positively correlated in DLBCL cell lines and primary DLBCL. Expression levels of FOXM1 and PLK1 mRNAs were also significantly higher in DLBCL than in normal human B cells and could predict poor prognosis of DLBCL, particularly in patients with germinal center B cell-like (GCB) DLBCL. Furthermore, proteomic studies defined a FOXM1-PLK1 signature that consisted of proteins upstream and downstream of that axis involved in the p38-MAPK-AKT pathway, cell cycle, and DNA damage/repair. Further studies demonstrated a mechanistic function of the FOXM1/PLK1 axis in connection with the DNA damage response pathways regulating the S/G2 checkpoint of the cell cycle. Therapeutic targeting of FOXM1/PLK1 using a FOXM1 or PLK1 inhibitor, as well as other clinically relevant small-molecule inhibitors targeting ATR-CHK1, was highly effective in DLBCL in vitro models. These findings are instrumental for lymphoma drug discovery aiming at the FOXM1/PLK1/ATR/CHK1 axis.

18.
Vet Sci ; 9(8)2022 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-35893772

RESUMO

Mastitis is one of the most widespread diseases in dairy cows and causes huge losses for the dairy industry. Molecular markers can be used for the quick diagnosis of mastitis infection, consequently reducing the loss caused by this disease. Lactoferrin (LTF) and Toll-like receptor 2 (TLR2) have been suggested as candidate genes for mastitis; however, their associations with the mastitis incidence and milk components have not been reported in Vietnamese Holstein cows. This study examined the association of TLR2 and LTF polymorphisms with subclinical mastitis and milk components in the Holstein breed raised in Vietnam. Among 192 samples, we identified 44 mastitis-positive samples (22.92%). The mastitis significantly reduced the fat and lactose components in milk (p < 0.001) but increased the protein concentration in milk. A total of 94 (49%) and 98 (51%) cows had AA and AB genotypes for the LTF gene, respectively. No significant association was found between the LTF genotypes and the milk component traits or mastitis incidence (p > 0.05). The interaction between LTF and mastitis incidence was significantly associated with the protein percentage (p = 0.01). A total of 78, 76, and 38 cows had genotypes GG, GT, and TT for the TLR2 gene, respectively. TLR2 genotypes were not significantly associated with mastitis incidence (p > 0.05) but were significantly associated with pH value (p = 0.03). The interaction between TLR2 and mastitis incidence was significantly associated with the fat (p = 0.02) and protein percentage (p = 0.04). Further studies are required to confirm the roles of LTF and TFL2 in mastitis in the Holstein breed in Vietnam.

19.
Sci Rep ; 12(1): 10485, 2022 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-35729348

RESUMO

The Vietnamese native pig (VnP)-a porcine breed with a small body-has proven suitable as a biomedical animal model. Here, we demonstrate that, compared to other breeds, VnPs have fewer copies of porcine endogenous retroviruses (PERVs), which pose a risk for xenotransplantation of pig organs to humans. More specifically, we sought to characterize non-reference PERVs (nrPERVs) that were previously unidentified in the reference genome. To this end, we used whole-genome sequencing data to identify nrPERV loci with long terminal repeat (LTR) sequences in VnPs. RetroSeq was used to estimate nrPERV loci based on the most current porcine reference genome (Sscrofa11.1). LTRs were detected using de novo sequencing read assembly near the loci containing the target site duplication sequences in the inferred regions. A total of 21 non-reference LTR loci were identified and separated into two subtypes based on phylogenetic analysis. Moreover, PERVs within the detected LTR loci were identified, the presence of which was confirmed using conventional PCR and Sanger sequencing. These novel loci represent previously unknown PERVs as they have not been identified in the porcine reference genome. Thus, our RetroSeq method accurately detects novel PERV loci, and can be applied for development of a useful biomedical model.


Assuntos
Retrovirus Endógenos , Gammaretrovirus , Animais , Povo Asiático , Retrovirus Endógenos/genética , Gammaretrovirus/genética , Humanos , Filogenia , Suínos/genética , Sequências Repetidas Terminais/genética , Transplante Heterólogo
20.
J Org Chem ; 87(9): 5952-5960, 2022 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-35417151

RESUMO

A voltammetric study of a series of alkyl and aryl S-glucosides unveiled the reactivity patterns of alkyl S-glucosides toward anodic oxidation and found noteworthy differences with the trends followed by aryl derivatives. The oxidation potential of alkyl S-glucosides, estimated herein from square-wave voltammetry peak potentials (Ep), depends on the steric properties of the aglycone. Glucosides substituted with bulky groups exhibit Ep values at voltages more positive than the values of those carrying small aglycones. This relationship, observed in all analyzed alkyl series, is evidenced by good linear correlations between Ep and Taft's steric parameters (ES) of the respective alkyl substituents. Moreover, the role of the aglycone's steric properties as a primary reactivity modulator is backed by poor correlations between Ep and the radical stabilization energies (RSEs) of the aglycone-derived thiyl radicals (RS•). In contrast, aryl glucosides' Ep values exhibit excellent correlations with the aryl substituents' Hammett parameters (σ+) and the ArS• RSEs, evidencing the inherent stability of the reactive radical intermediate as the primary factor controlling aryl glucoside's electrochemical reactivity. The reactivity differences between alkyl and aryl S-glucosides also extend to the protective group's effect on Ep. Alkyl S-glucosides' reactivity proved to be more sensitive to protective group exchange.


Assuntos
Glucosídeos , Eletrodos , Oxirredução
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