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1.
Anal Chim Acta ; 1186: 339117, 2021 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-34756250

RESUMO

Transferrin (Trf) is a new type of active drug targeting carrier and disease biomarker that regulates the balance of iron ions in human body. The recognition and isolation of Trf is of great significance for disease diagnosis and treatment. Thus, a new type of magnetic dual affinity epitope molecularly imprinted polymer coated on Fe3O4 nanoparticles (Fe3O4@DEMIP) was successfully prepared for specific recognition of Trf. C-terminal nonapeptide and Trf glycan were selected as bi-epitope templates for metal chelation and boron affinity immobilization, respectively. 4-vinylphenylboric acid (4-VP), N-isopropyl acrylamide (NIPAM) and zinc acrylic were used as functional monomers. Results showed that Fe3O4@DEMIP exhibited excellent specific recognition ability adsorption capacity toward Trf, with an adsorption of 43.96 mg g-1 (RSD = 3.28%) and a more satisfactory imprinting factor (about 6.60) than that of other reported imprinting methods. In addition, Fe3O4@DEMIP displayed pH, temperature and magnetic sensitivity properties to realize temperature and pH-controlled recognition and release of target proteins and magnetic rapid separation. Furthermore, the Fe3O4@DEMIP coupled with high-performance liquid chromatography (HPLC) analysis was successfully used for specific recognition of Trf in biosamples. This study provides a reliable protocol for preparing metal chelation and boron affinity dual affinity bi-epitope molecularly imprinted polymers for synergistic and efficient recognition of biomacromolecules in the complex biological systems.


Assuntos
Impressão Molecular , Polímeros , Adsorção , Epitopos , Humanos , Transferrina
2.
Zhonghua Fu Chan Ke Za Zhi ; 56(10): 665-670, 2021 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-34823314

RESUMO

Objective: To study the cut-off values of urinary microalbumin (mAlb), transferrin (TRF) and α1-microglobulin (α1-MG) during pregnancy in pre-eclampsia (PE) with proteinuria. Methods: A total of 210 pregnant women were enrolled in Renji Hospital from January 2016 to December 2019, including 92 (43.8%) cases of PE pregnant women and 118 (56.2%) cases of normal pregnant women. According to the diagnostic test evaluation method, the positive predictive values, negative predictive values and accuracy of non-pregnant cut-off values of urinary mAlb, TRF and α1-MG for the quantitative determination of 24-hour proteinuria were analyzed. The receiver operating characteristic (ROC) curve was applied to determine the optimal cut-point values of urinary mAlb, TRF and α1-MG during pregnancy. Results: (1) The diagnostic study of non-pregnant adults urinary mAlb, TRF and α1-MG cut-off values for the determination of 24-hour proteinuria value: when urinary mAlb was 30.0 mg/L, TRF was 2.5 mg/L, α1-MG was 12.5 mg/L as the cut-off value, the positive predictive values of the corresponding 24-hour proteinuria value≥ 300 mg were 88.1% (89/101), 88.2% (90/102) and 78.9% (75/95), its negative predictive values were 97.2% (106/109), 98.1% (106/108) and 85.2% (98/115), its diagnostic accuracy were 92.9% (195/210), 93.3% (196/210) and 82.4% (173/210), respectively. As the 24-hour proteinuria value≥ 300 mg was the golden standard, there were significant differences between the diagnostic method of the non-pregnant cut-off value of urinary mAlb, TRF and the golden standard (P<0.05). There was no significant difference between the diagnostic method of the non-pregnant cut-off value of urinary α1-MG and the golden standard (P>0.05). (2) Research on the ROC curve and the optimal cut-point value of urinary mAlb, TRF and α1-MG value: as the 24-hour proteinuria value≥ 300 mg as the criterion, the ROC curve of urinary mAlb, TRF and α1-MG were 0.992, 0.984 and 0.907, respectively. The optimal cut-point values of urinary mAlb, TRF and α1-MG were 86.5 mg/L (Youden index=0.927), 5.5 mg/L (Youden index=0.923), and 15.4 mg/L (Youden index=0.687). (3) The diagnostic study of the optimal cut-point value of urinary mAlb, TRF and α1-MG for the determination of 24-hour proteinuria value: according to the ROC results, when urinary mAlb was 86.5 mg/L, urinary TRF was 5.5 mg/L, and urinary α1-MG was 15.4 mg/L as the cut-off value, the positive predictive values of the corresponding 24-hour proteinuria value≥300 mg were 98.9% (86/87), 95.7% (88/92), 87.7% (71/81), and its negative predictive values were 95.1% (117/123), 96.6% (114/118), 83.7% (108/129), and its accuracy were 96.7% (203/210), 96.2% (202/210), 85.2% (179/210). As the 24-hour proteinuria value≥ 300 mg was the golden standard, there was no significant difference between the diagnostic method of the best cut-off values of urinary mAlb, TRF, α1-MG and the golden standard (P>0.05). Conclusion: It is recommended to define the cut-off values of mAlb, TRF and α1-MG as 86.5 mg/L, 5.5 mg/L and 15.4 mg/L, respectively, during pregnancy.


Assuntos
Pré-Eclâmpsia , Adulto , Feminino , Humanos , Pré-Eclâmpsia/diagnóstico , Valor Preditivo dos Testes , Gravidez , Proteinúria/diagnóstico , Curva ROC , Sensibilidade e Especificidade , Transferrina
3.
Sci Rep ; 11(1): 19618, 2021 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-34608227

RESUMO

The pathophysiology and the factors determining disease severity in COVID-19 are not yet clear, with current data indicating a possible role of altered iron metabolism. Previous studies of iron parameters in COVID-19 are cross-sectional and have not studied catalytic iron, the biologically most active form of iron. The study was done to determine the role of catalytic iron in the adverse outcomes in COVID-19. We enrolled adult patients hospitalized with a clinical diagnosis of COVID-19 and measured serum iron, transferrin saturation, ferritin, hepcidin and serum catalytic iron daily. Primary outcome was a composite of in-hospital mortality, need for mechanical ventilation, and kidney replacement therapy. Associations between longitudinal iron parameter measurements and time-to-event outcomes were examined using a joint model. We enrolled 120 patients (70 males) with median age 50 years. The primary composite outcome was observed in 25 (20.8%) patients-mechanical ventilation was needed in 21 (17.5%) patients and in-hospital mortality occurred in 21 (17.5%) patients. Baseline levels of ferritin and hepcidin were significantly associated with the primary composite outcome. The joint model analysis showed that ferritin levels were significantly associated with primary composite outcome [HR (95% CI) = 2.63 (1.62, 4.24) after adjusting for age and gender]. Both ferritin and serum catalytic iron levels were positively associated with in-hospital mortality [HR (95% CI) = 3.22 (2.05, 5.07) and 1.73 (1.21, 2.47), respectively], after adjusting for age and gender. The study shows an association of ferritin and catalytic iron with adverse outcomes in COVID-19. This suggests new pathophysiologic pathways in this disease, also raising the possibility of considering iron chelation therapy.


Assuntos
COVID-19/patologia , Ferro/sangue , Adulto , Idoso , COVID-19/mortalidade , COVID-19/virologia , Estudos Transversais , Feminino , Ferritinas/sangue , Ferritinas/metabolismo , Hepcidinas/sangue , Hepcidinas/metabolismo , Mortalidade Hospitalar , Humanos , Ferro/química , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Respiração Artificial , SARS-CoV-2/isolamento & purificação , Índice de Gravidade de Doença , Transferrina/química , Transferrina/metabolismo
4.
Am Fam Physician ; 104(3): 263-270, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34523883

RESUMO

Hereditary hemochromatosis is an autosomal recessive disorder that disrupts iron homeostasis, resulting in systemic iron overload. It is the most common inherited disorder among people of northern European ancestry. Despite the high prevalence of the gene mutation, there is a low and variable clinical penetrance. The deposition of excess iron into parenchymal cells leads to cellular dysfunction and the clinical manifestations of the disease. The liver, pancreas, joints, heart, skin, and pituitary gland are the most commonly involved organs. Hereditary hemochromatosis is usually diagnosed in the 40s or 50s. Women are often diagnosed later than men, likely because of menstrual blood loss. There is no typical presentation or pathognomonic signs and symptoms of hereditary hemochromatosis. Because of increased awareness and earlier diagnosis, the end-organ damage secondary to iron overload is not often seen in clinical practice. A common initial presentation is an asymptomatic patient with mildly elevated liver enzymes who is subsequently found to have elevated serum ferritin and transferrin saturation. Ferritin levels greater than 300 ng per mL for men and 200 ng per mL for women and transferrin saturations greater than 45% are highly suggestive of hereditary hemochromatosis. Phlebotomy is the mainstay of treatment and can help improve heart function, reduce abnormal skin pigmentation, and lessen the risk of liver complications. Liver transplantation may be considered in select patients. Individuals with hereditary hemochromatosis have an increased risk of hepatocellular carcinoma and colorectal and breast cancers. Genetic testing for the hereditary hemochromatosis genes should be offered after 18 years of age to first-degree relatives of patients with the condition.


Assuntos
Hemocromatose/diagnóstico , Hemocromatose/epidemiologia , Hemocromatose/fisiopatologia , Humanos , Cirrose Hepática/sangue , Cirrose Hepática/etiologia , Programas de Rastreamento/métodos , Transferrina/análise
5.
Clin Lab ; 67(9)2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34542961

RESUMO

BACKGROUND: We aimed to evaluate the use of population-based-reference-intervals by calculating the individuality index and reference change value of iron, transferrin, ferritin, folate, vitamin B12, and 25-OH vitamin D parameters. METHODS: A total of 5 venous blood samples were taken from 11 female and 10 male individuals on days 0, 7, 14, 21, and 28. All tests were performed on an autoanalyzer and CVI, CVG, II, and RCV values were calculated for both genders and the whole groups. RESULTS: CVA (%)/CVI (%)/CVG (%) for iron, 0.67/27.3/32.3 transferrin, 0.62/3.60/10.27 ferritin, 2.27/6.21/105.6 folate 4.71/10.3/28.56 vitamin B12, 6.1/5.77/34.6, 25-OH Vitamin D 3.4/8.2/54.9 respectively. RCV calculated as a 2-tailed value at level of probability of significant change set at 0.95 - 0.99; 74.9/98.7 - 10.1/13.3 - 18.3/24.1 - 31.4/41.3 - 23.4/30.8 - 24.7/32.5 and II were 0.8-0.4-0.-0.4-0.17-0.1 respectively. CONCLUSIONS: CVI and CVG values for iron, transferrin, folate, vitamin B12, and 25-OH vitamin D are in accordance with the literature. Ferritin was calculated differently from the values in the database. This difference might be due to the characteristics of selected individuals for study. Evaluating the suitability and utility of the use of RR with values found for II. For iron, the use of RR was considered appropriate when taken as II < 0.6 and II > 1.4. However, the use of RCV is more appropriate for Transferrin, Folate, Vitamin B12, and 25-OH Vitamin D.


Assuntos
Ferritinas , Vitamina B 12 , Feminino , Ácido Fólico , Humanos , Ferro , Masculino , Transferrina , Vitamina D
6.
Chem Commun (Camb) ; 57(80): 10391-10394, 2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34542119

RESUMO

Transferrin-modified AuNCs (Tf-AuNCs) with two photon-near infrared (TP-NIR) fluorescence were prepared. For the first time, a novel nanoprobe platform, Tf-AuNCs@MnO2, was developed for the TP-NIR fluorescence imaging and magnetic resonance imaging of living cells and tissues. This platform had high spatiotemporal resolution and a tissue-penetration depth of 300 µm.


Assuntos
Corantes Fluorescentes/química , Nanopartículas Metálicas/química , Transferrina/química , Corantes Fluorescentes/efeitos da radiação , Fluorometria , Glutationa/análise , Glutationa/metabolismo , Ouro/química , Ouro/efeitos da radiação , Humanos , Raios Infravermelhos , Células MCF-7 , Compostos de Manganês/química , Nanopartículas Metálicas/efeitos da radiação , Óxidos/química , Fótons
7.
Artigo em Inglês | MEDLINE | ID: mdl-34562778

RESUMO

Transferrin is a glycoprotein containing two bi- or tri-antennary carbohydrate chains ending with sialic acid. Its glycosylation is reduced in chronic alcohol abuse and in inborn glycosylation pathologies, where the carbohydrate-deficient fraction of the protein (CDT) increases significantly. The current methods require a gradient chromatographic separation and time-consuming sample preparation. In comparison, the proposed approach uses a novel flow-modulated liquid chromatography technique (fmLC) and a highly selective and sensitive fluorescence derivatization reaction with terbium ion. A fmLC-FLD method using isocratic anion exchange separation was optimized and validated to resolve disialo-transferrin and trisialo-transferrin from other transferrin glycoforms. Detection took place by recording fluorescence at 550 nm wavelength (excitation at 298 nm). The chromatographic separation needed 5 min, allowing seriate injection every 7.5 min. The method was validated according to the current guidelines of analytical chemistry showing adequate accuracy and precision for the quantitative determination of CDT. The proposed method proved also to be suitable to analyse haemolyzed sera which, because of interference by haemoglobin, fail the standard HPLC-Vis analysis. The method was tested in parallel with HPLC-Vis on 131 sera showing an excellent correlation of results proved by a correlation coefficient of 0.995 (Pearson's r). The proposed approach proved much simpler than the current methods and cheaper in terms of instrumental costs offering a ground-breaking analytical tool that could likely make available the characterization of CDT outside specialized laboratories, such as in occupational medicine centres, doctor's offices, small laboratories, alcohol rehabilitation centres, and in developing countries.


Assuntos
Cromatografia Líquida/métodos , Espectrometria de Fluorescência/métodos , Transferrina/análogos & derivados , Alcoolismo , Biomarcadores , Ensaios de Triagem em Larga Escala , Humanos , Limite de Detecção , Reprodutibilidade dos Testes , Térbio/química , Transferrina/análise
8.
AAPS PharmSciTech ; 22(7): 239, 2021 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-34590204

RESUMO

The purpose of this study was to enhance the antitumor effect of piperine by constructing the nanoparticles modified with transferrin (Tf-PIP-NPs) and evaluating their efficacy in vitro and in vivo. The Tf-PIP-NPs were prepared by the solvent evaporation method, and their properties were characterized. The effects of Tf-PIP-NPs on cytotoxicity, cell uptake, apoptosis, and mitochondrial membrane potential were evaluated in HepG2 cells, MDA-MB-231 cells, and 4T1 cells. In a 4T1 tumor-bearing mouse model, the antitumor efficacy of Tf-PIP-NPs was assessed in terms of tumor volumes, changes in body weight, HE staining, and immunohistochemical analysis. With a mean particle size of 112.2 ± 1.27 nm, the zeta potential of (- 28.0 ± 1.6 mV) Tf-PIP-NPs were rapidly internalized by tumor cells after 1 h through the transferrin receptor (TfR)-mediated endocytosis pathway, significantly inducing cellular apoptosis and mitochondrial membrane potential loss. Although Tf-PIP-NPs had no significant difference with PIP-NPs in tumor volume inhibition due to the presence of tumor microenvironment, it could significantly upregulate the expression of related pro-apoptotic proteins and induce tumor necrosis. We used the self-assembly properties of glycyrrhizic acid (GL) and polymer-PLGA to encapsulate piperine and modified with the transferrin, which provided a promising approach to improve the antitumor efficacy for anticarcinogen.


Assuntos
Nanopartículas , Transferrina , Alcaloides , Animais , Benzodioxóis , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Ácido Glicirrízico , Camundongos , Tamanho da Partícula , Piperidinas , Alcamidas Poli-Insaturadas
10.
J Insect Sci ; 21(4)2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-34401920

RESUMO

Glycosylation is one of the most common post-translational modifications to occur during protein biosynthesis, but remains poorly understood in insects. In this study, we collected serum proteins from two silkworm developmental stages, namely day 7 of the fifth instar larval stage and day 2 of the pupal stage. Results of SDS-PAGE and periodic acid-Schiff staining revealed that most serum proteins with high abundance were putative glycoproteins. LC-MS/MS identified 149 larval and 303 pupal serum proteins in the Con A lectin-enriched fractions. GO analysis revealed that many serum proteins were involved in the proteolysis and carbohydrate metabolic process. 82 N-linked glycoproteins with at least one glycosylation site were identified. N-Linked glycosylation occurred at the sequon, Asn-X-Ser/Thr, and the proportions of Ser and Thr glycosylation at the hydroxy position were found 39.6% and 60.3%, respectively. The N-glycan structures found in serum glycoproteins were mainly Man2FucGlcNAc2 (67.9%). Since storage protein 1 and transferrin had a relatively high abundance in the serum and could be significantly enriched by Con A lectin, their glycosylation was analyzed in detail. Glycoside hydrases, serine proteases and serpins were found to form three interacting glycoprotein networks using the website STRING. This study provides important clues for the understanding of the function of N-linked glycosylation in metabolism, immunity, and metamorphosis.


Assuntos
Bombyx/metabolismo , Glicoproteínas/metabolismo , Receptores de Concanavalina A/metabolismo , Animais , Cromatografia de Afinidade , Glicosilação , Proteínas de Insetos/metabolismo , Espectrometria de Massas , Proteômica , Transferrina/metabolismo
11.
Fish Shellfish Immunol ; 117: 169-178, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34389379

RESUMO

It is known that iron transporter proteins and their regulation can modulate the fish's immune system, suggesting these proteins as a potential candidate for fish vaccines. Previous studies have evidenced the effects of Atlantic salmon immunized with the chimeric iron-related protein named IPath® against bacterial and ectoparasitic infections. The present study aimed to explore the transcriptome modulation and the morphology of the sea louse Caligus rogercresseyi in response to Atlantic salmon injected with IPath®. Herein, Atlantic salmon were injected with IPath® and challenged to sea lice in controlled laboratory conditions. Then, female adults were collected after 25 days post-infection for molecular and morphological evaluation. Transcriptome analysis conducted in lice collected from immunized fish revealed high modulation of transcripts compared with the control groups. Notably, the low number of up/downregulated transcripts was mainly found in lice exposed to the IPath® fish group. Among the top-25 differentially expressed genes, Vitellogenin, Cytochrome oxidases, and proteases genes were strongly downregulated, suggesting that IPath® can alter lipid transport, hydrogen ion transmembrane transport, and proteolysis. The morphological analysis in lice collected from IPath® fish revealed abnormal embryogenesis and inflammatory processes of the genital segment. Furthermore, head kidney, spleen, and skin were also analyzed in immunized fish to evaluate the transcription expression of immune and iron homeostasis-related genes. The results showed downregulation of TLR22, MCHII, IL-1ß, ALAs, HO, BLVr, GSHPx, and Ferritin genes in head kidney and skin tissues; meanwhile, those genes did not show significant differences in spleen tissue. Overall, our findings suggest that IPath® can be used to enhance the fish immune response, showing a promissory commercial application against lice infections.


Assuntos
Copépodes/genética , Ectoparasitoses/prevenção & controle , Doenças dos Peixes/prevenção & controle , Proteínas Recombinantes/administração & dosagem , Salmo salar/parasitologia , Transcriptoma , Vacinas/administração & dosagem , Animais , Ectoparasitoses/veterinária , Feminino , Ferritinas/genética , Salmo salar/imunologia , Transferrina/genética , Vacinação
12.
Blood Adv ; 5(16): 3102-3112, 2021 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-34402883

RESUMO

Acute myeloid leukemia (AML) is a heterogeneous disease with poor prognosis and limited treatment strategies. Determining the role of cell-extrinsic regulators of leukemic cells is vital to gain clinical insights into the biology of AML. Iron is a key extrinsic regulator of cancer, but its systemic regulation remains poorly explored in AML. To address this question, we studied iron metabolism in patients with AML at diagnosis and explored the mechanisms involved using the syngeneic MLL-AF9-induced AML mouse model. We found that AML is a disorder with a unique iron profile, not associated with inflammation or transfusion, characterized by high ferritin, low transferrin, high transferrin saturation (TSAT), and high hepcidin. The increased TSAT in particular, contrasts with observations in other cancer types and in anemia of inflammation. Using the MLL-AF9 mouse model of AML, we demonstrated that the AML-induced loss of erythroblasts is responsible for iron redistribution and increased TSAT. We also show that AML progression is delayed in mouse models of systemic iron overload and that elevated TSAT at diagnosis is independently associated with increased overall survival in AML. We suggest that TSAT may be a relevant prognostic marker in AML.


Assuntos
Anemia , Leucemia Mieloide Aguda , Animais , Eritroblastos , Humanos , Ferro , Camundongos , Transferrina
13.
Nutrients ; 13(7)2021 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-34371810

RESUMO

Ferrous sulphate (FS) is a cost effective, readily available iron supplement for iron deficiency (ID). The pro-oxidant effect of oral ferrous iron is known to induce inflammation, causing gastric side-effects and resulting in poor compliance. Curcumin is a potent antioxidant and has also been shown to exhibit iron chelation in-vitro, although it is not established whether these effects are retained in-vivo. The aim of this study was therefore to assess the influence of a formulated bioavailable form of curcumin (HydroCurcTM; 500 mg) on acute iron absorption and status in a double blind, placebo-controlled randomized trial recruiting 155 healthy participants (79 males; 26.42 years ± 0.55 and 76 females; 25.82 years ± 0.54). Participants were randomly allocated to five different treatment groups: iron and curcumin placebo (FS0_Plac), low dose (18 mg) iron and curcumin placebo (FS18_Plac), low dose iron and curcumin (FS18_Curc), high dose (65 mg) iron and curcumin placebo (FS65_Plac), and high dose iron and curcumin (FS65_Curc). Participants were provided with the supplements according to their relevant treatment groups at baseline (0 min), and blood collection was carried out at 0 min and at 180 min following supplementation. In the treatment groups, significant difference was observed in mean serum iron between baseline (0 min) and at end-point (180 min) (F (1, 144) = 331.9, p < 0.0001) with statistically significant intra-group increases after 180 min (p < 0.0001) in the FS18_Plac (8.79 µmol/L), FS18_Curc (11.41 µmol/L), FS65_Plac (19.09 µmol/L), and FS65_Curc (16.39 µmol/L) groups. A significant difference was also observed between the two time points in serum TIBC levels and in whole blood haemoglobin (HGB) in the treatment groups, with a significant increase (1.55%/2.04 g/L) in HGB levels from baseline to end-point observed in the FS65_Curc group (p < 0.05). All groups receiving iron demonstrated an increase in transferrin saturation (TS%) in a dose-related manner, demonstrating that increases in serum iron are translated into increases in physiological iron transportation. This study demonstrates, for the first time, that regardless of ferrous dose, formulated curcumin in the form of HydroCurc™ does not negatively influence acute iron absorption in healthy humans.


Assuntos
Absorção Fisiológica/efeitos dos fármacos , Curcumina/administração & dosagem , Suplementos Nutricionais , Compostos Ferrosos/administração & dosagem , Ferro/sangue , Administração Oral , Adulto , Disponibilidade Biológica , Método Duplo-Cego , Feminino , Ferritinas/sangue , Voluntários Saudáveis , Hemoglobinas/análise , Humanos , Proteínas de Ligação ao Ferro/sangue , Masculino , Transferrina/análise
14.
Fukushima J Med Sci ; 67(2): 64-70, 2021 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-34373399

RESUMO

Spontaneous intracranial hypotension (SIH) is caused by cerebrospinal fluid (CSF) leakage. Patients with SIH experience postural headaches, nausea, etc., due to CSF hypovolemia. Imaging studies and clinical examinations, such as radioisotope (RI) scintigraphy, are useful for diagnosing SIH. However, 20-30% of patients do not show typical morphology and clinical test results. We previously reported that CSF contains transferrin (Tf) isoforms:"brain-type" Tf derived from the choroid plexus and "serum-type" Tf derived from blood. We showed that both isoforms increased in the CSF of patients with SIH by Western blotting. In the present study, we demonstrate that conventional ELISA for quantifying total Tf is useful for diagnosing SIH more accurately than Western blotting. In addition, SIH with chronic subdural hematoma (CSDH) was also accurately diagnosed. Total Tf in the CSF can serve as a useful biomarker for diagnosing SIH with or without CSDH.


Assuntos
Hipotensão Intracraniana , Biomarcadores , Encéfalo , Vazamento de Líquido Cefalorraquidiano/diagnóstico , Humanos , Hipotensão Intracraniana/diagnóstico , Transferrina
15.
Int J Pharm ; 607: 121034, 2021 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-34425193

RESUMO

Our previous studies have proven that carnosic acid (CA) induces apoptosis of liver cancer cells. However, the poor chemical properties of CA limit its in vivo anti-cancer effects. In this study, CA was loaded into liposomes (LP-CA), and LP-CA was further conjugated with transferrin (Tf-LP-CA) to overcome the shortcomings of poor solubility and absorption at the lesion site. In HepG2 and SMMC-7721 cells, compared with CA and LP-CA, more Tf-LP-CA was absorbed by liver cancer cells, which induced higher levels of apoptosis and reduced the mitochondrial membrane potential more effectively. In HepG2- and SMMC-7721-xenotransplanted mice, Tf-LP-CA inhibited tumor growth with no cytotoxicity to the liver, spleen, or kidney. Furthermore, compared with CA and LP-CA, Tf-LP-CA targeted the tumor site more effectively, enhanced the expressions of cleaved poly(ADP-ribose) polymerase, and Caspase-3 and -9, and regulated the expression levels of B-cell lymphoma 2 (Bcl2) family members in the tumor tissues. Tf-LP-CA was taken up by tumor cells and targeted at tumor tissues, ensuring the precise delivery of CA, which further promoted mitochondria-mediated intrinsic apoptosis in the liver cancer cells. These results provide evidence for the clinical application of the Tf-LP-based CA drug delivery system for liver cancer.


Assuntos
Lipossomos , Neoplasias Hepáticas , Abietanos , Animais , Apoptose , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Neoplasias Hepáticas/tratamento farmacológico , Camundongos , Mitocôndrias , Transferrina
16.
Int J Pharm ; 608: 121038, 2021 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-34438008

RESUMO

New drug discovery and development processes encounter significant challenges including requirement of huge investments and lengthy time frames especially in cancer research field. Repurposing of old drugs against cancer provides a possible alternative while associated scale-up complexities with production of nanoparticles at industrial scale could be overcome by using a scalable nanoparticle technique. We previously described use of polymeric nanoparticles for inhaled delivery of amodiaquine (AQ) for non-small cell lung cancer (NSCLC) treatment. In this study, targeting potential of transferrin ligand conjugated inhalable AQ-loaded nanoparticles (Tf-AMQ NPs) was investigated against NSCLC. Tf-AMQ NP (liquid formulation) demonstrated an aerodynamic diameter of 4.4 ± 0.1 µm and fine particle fraction of 83.2 ± 3.0%, representing AQ deposition in the respirable region of airways. Cytotoxicity studies in NSCLC cell line with overexpressed transferrin receptors shown significant reduction in IC50 values with Tf-decorated AQ-loaded nanoparticles compared to AQ or non-targeted NPs, along with significant apoptosis induction (caspase assay) and reduced % colony growth in A549 and H1299 cells with Tf-AMQ NP. Furthermore, 3D spheroid studies (~7-fold reduction in spheroid volume compared to AMQ NPs) explained efficiency of conjugated nanoparticles in penetrating tumor core, and growth inhibition. AQ's autophagy inhibition ability significantly increased with nanoparticle encapsulation and transferrin conjugation. In conclusion, amodiaquine can be an assuring candidate for repurposing to consider for NSCLC treatment while delivering inhalable transferrin conjugated nanoparticles developed using a scalable HPH process to the target site, thus reducing the dose, side effects.


Assuntos
Amodiaquina , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Nanopartículas , Células A549 , Amodiaquina/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Transferrina
17.
J Control Release ; 337: 521-529, 2021 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-34352315

RESUMO

Brain metastases are a most disturbing situation for breast cancer patients as there is basically no adequate treatment available. Any potential drug formulation has to be able to cross the blood-brain barrier (BBB) and specific to metastatic brain tumors without causing unacceptable adverse effects. Here, we developed transferrin-functionalized chimeric polymersomes carrying siRNA against polo-like kinase 1 (Tf@TBP-CPs-siPLK1) for treating brain metastatic MDA-MB 231 triple negative breast cancer (TNBC) xenografts in mice. To facilitate the loading of siPLK1, chimaeric polymersomes (CPs) were designed with spermine in the watery core and transferrin-binding peptide (TBP) at the surface, enabling attachment of transferrin after the siRNA loading step and thereby circumventing interference of transferrin with siRNA loading. Tf@TBP-CPs-siPLK1 encapsulating 3.8 wt% siRNA had a mean size of about 50 nm and a neutral zeta potential in phosphate buffer (PB). By virtue of the presence of transferrin, Tf@TBP-CPs demonstrated greatly (ca. 5-fold) enhanced internalization in MDA-MB 231 cells and transcytosis in the endothelial (bEnd.3) monolayer model in vitro as well as markedly improved accumulation in the orthotopically xenografted MDA-MB 231 tumor in the brain in vivo compared with control CPs lacking transferrin, supporting that transferrin mediates efficient BBB penetration and high specificity towards MDA-MB 231 cells. As a result, Tf@TBP-CPs-siPLK1 effectively inhibited tumor progression and prolonged the lifespan of the mice significantly. Selective transferrin coating appears to be a particularly facile strategy to fabricate BBB-permeable and targeted vesicles for potent RNAi therapy of brain metastatic breast cancer.


Assuntos
Neoplasias Encefálicas , Neoplasias da Mama , Animais , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Neoplasias Encefálicas/terapia , Neoplasias da Mama/terapia , Linhagem Celular Tumoral , Feminino , Humanos , Camundongos , Terapêutica com RNAi , Transferrina/metabolismo
18.
Cells ; 10(7)2021 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-34359837

RESUMO

Induced pluripotent stem (iPS) cells constitute a perfect tool to study human embryo development processes such as myogenesis, thanks to their ability to differentiate into three germ layers. Currently, many protocols to obtain myogenic cells have been described in the literature. They differ in many aspects, such as media components, including signaling modulators, feeder layer constituents, and duration of culture. In our study, we compared three different myogenic differentiation protocols to verify, side by side, their efficiency. Protocol I was based on embryonic bodies differentiation induction, ITS addition, and selection with adhesion to collagen I type. Protocol II was based on strong myogenic induction at the embryonic bodies step with BIO, forskolin, and bFGF, whereas cells in Protocol III were cultured in monolayers in three special media, leading to WNT activation and TGF-ß and BMP signaling inhibition. Myogenic induction was confirmed by the hierarchical expression of myogenic regulatory factors MYF5, MYOD, MYF6 and MYOG, as well as the expression of myotubes markers MYH3 and MYH2, in each protocol. Our results revealed that Protocol III is the most efficient in obtaining myogenic cells. Furthermore, our results indicated that CD56 is not a specific marker for the evaluation of myogenic differentiation.


Assuntos
Técnicas de Cultura de Células , Meios de Cultura/farmacologia , Corpos Embrioides/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Desenvolvimento Muscular/efeitos dos fármacos , Fibras Musculares Esqueléticas/efeitos dos fármacos , Biomarcadores/metabolismo , Diferenciação Celular/efeitos dos fármacos , Colforsina/farmacologia , Colágeno Tipo I/farmacologia , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Corpos Embrioides/citologia , Corpos Embrioides/metabolismo , Fator 2 de Crescimento de Fibroblastos/farmacologia , Fibroblastos/citologia , Fibroblastos/metabolismo , Expressão Gênica , Humanos , Indóis/farmacologia , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Insulina/farmacologia , Desenvolvimento Muscular/genética , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/metabolismo , Proteína MyoD/genética , Proteína MyoD/metabolismo , Fator Regulador Miogênico 5/genética , Fator Regulador Miogênico 5/metabolismo , Fatores de Regulação Miogênica/genética , Fatores de Regulação Miogênica/metabolismo , Miogenina/genética , Miogenina/metabolismo , Oximas/farmacologia , Selênio/farmacologia , Transferrina/farmacologia
19.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34210014

RESUMO

Human serum transferrin (HST) is a glycoprotein involved in iron transport that may be a candidate for functionalized nanoparticles to bind and target cancer cells. In this study, the effects of the simple and doped with cobalt (Co) and copper (Cu) ferrihydrite nanoparticles (Fh-NPs, Cu-Fh-NPs, and Co-Fh-NPs) were studied by spectroscopic and molecular approaches. Fluorescence spectroscopy revealed a static quenching mechanism for all three types of Fh-NPs. All Fh-NPs interacted with HST with low affinity, and the binding was driven by hydrogen bonding and van der Waals forces for simple Fh-NPs and by hydrophobic interactions for Cu-Fh-NPs and Co-Fh-NPs binding, respectively. Of all samples, simple Fh-NPs bound the most to the HST binding site. Fluorescence resonance energy transfer (FRET) allowed the efficient determination of the energy transfer between HST and NPs and the distance at which the transfer takes place and confirmed the mechanism of quenching. The denaturation of the HST is an endothermic process, both in the case of apo HST and HST in the presence of the three types of Fh-NPs. Molecular docking studies revealed that Fh binds with a low affinity to HST (Ka = 9.17 × 103 M-1) in accord with the fluorescence results, where the interaction between simple Fh-NPs and HST was described by a binding constant of 9.54 × 103 M-1.


Assuntos
Cobalto/química , Compostos Férricos/síntese química , Transferrina/química , Transferrina/metabolismo , Cobre/química , Compostos Férricos/química , Transferência Ressonante de Energia de Fluorescência , Humanos , Ligação de Hidrogênio , Modelos Moleculares , Simulação de Acoplamento Molecular , Nanopartículas , Ligação Proteica , Conformação Proteica , Espectrometria de Fluorescência , Termodinâmica
20.
Int J Mol Sci ; 22(11)2021 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-34199599

RESUMO

Orphan nuclear receptor estrogen-related receptor γ (ERRγ) is an important transcription factor modulating gene transcription involved in endocrine control of liver metabolism. Transferrin receptor 2 (TFR2), a carrier protein for transferrin, is involved in hepatic iron overload in alcoholic liver disease (ALD). However, TFR2 gene transcriptional regulation in hepatocytes remains largely unknown. In this study, we described a detailed molecular mechanism of hepatic TFR2 gene expression involving ERRγ in response to an endocannabinoid 2-arachidonoylglycerol (2-AG). Treatment with 2-AG and arachidonyl-2'-chloroethylamide, a selective cannabinoid receptor type 1 (CB1) receptor agonist, increased ERRγ and TFR2 expression in hepatocytes. Overexpression of ERRγ was sufficient to induce TFR2 expression in both human and mouse hepatocytes. In addition, ERRγ knockdown significantly decreased 2-AG or alcohol-mediated TFR2 gene expression in cultured hepatocytes and mouse livers. Finally, deletion and mutation analysis of the TFR2 gene promoter demonstrated that ERRγ directly modulated TFR2 gene transcription via binding to an ERR-response element. This was further confirmed by chromatin immunoprecipitation assay. Taken together, these results reveal a previously unrecognized role of ERRγ in the transcriptional regulation of TFR2 gene expression in response to alcohol.


Assuntos
Hepatopatias Alcoólicas/genética , Fígado/efeitos dos fármacos , Receptor CB1 de Canabinoide/genética , Receptores de Estrogênio/genética , Receptores da Transferrina/genética , Álcoois/farmacologia , Animais , Ácidos Araquidônicos/farmacologia , Endocanabinoides/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Glicerídeos/farmacologia , Células Hep G2 , Hepatócitos/efeitos dos fármacos , Humanos , Ferro/metabolismo , Fígado/metabolismo , Fígado/patologia , Hepatopatias Alcoólicas/metabolismo , Hepatopatias Alcoólicas/patologia , Camundongos , Regiões Promotoras Genéticas , Receptor CB1 de Canabinoide/agonistas , Deleção de Sequência/genética , Transferrina/genética , Transferrina/metabolismo
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