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1.
Sci Rep ; 12(1): 2855, 2022 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-35190572

RESUMO

Uncontrollable proliferation is a hallmark of cancer cells. Cell proliferation and migration are significantly depressed during hibernation state. Many studies believe some factors in the plasma of hibernating animals cause these effects. This study aimed to assess the anti-cancer effects of hibernating common carp (Cyprinus carpio) plasma on 4T1 cancer cells in vitro and in vivo. The effect of hibernating plasma on cell viability, morphology, migration, apoptosis rate, and cell cycle distribution of 4T1 cells was investigated in vitro and in vivo. Hibernating plasma at a concentration of 16 mg/ml significantly reduced the viability of 4T1 cancer cells, without any toxicity on L929 normal fibroblast cells. It could change the morphology of cancer cells, induced apoptosis and cell cycle arrest at the G2/M phase, and inhibited migration. Furthermore, intratumoral injection of hibernating plasma (200 µl, 16 mg/ml) in the tumor-bearing mice caused a significant inhibition of 4T1 breast tumors volume (46.9%) and weight (58.8%) compared with controls. A significant decrease in the number of metastatic colonies at the lungs (80%) and liver (52.8%) of hibernating plasma-treated animals was detected which increased the survival time (21.9%) compared to the control groups. Immunohistochemical analysis revealed a considerable reduction in the Ki-67-positive cells in the tumor section of the hibernating plasma-treated animals compared with controls. Taken together, the SDS-PAGE and mass spectrometry analysis indicated the alpha-2-macroglobulin level in the hibernating fish plasma was significantly increased. It could exert an anti-cancer effect on breast cancer cells and suggested as a novel cancer treatment strategy.


Assuntos
Antineoplásicos/farmacologia , Carpas , Hibernação , Plasma/química , Plasma/fisiologia , Neoplasias de Mama Triplo Negativas/patologia , Animais , Antineoplásicos/isolamento & purificação , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Camundongos , Neoplasias de Mama Triplo Negativas/tratamento farmacológico
2.
Biotechnol Lett ; 43(6): 1143-1154, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33751317

RESUMO

Ansamitocin (AP-3) is an ansamycins antibiotic isolated from Actinosynnema pretiosum and demonstrating high anti-tumor activity. To improve AP-3 production, the A. pretiosum ATCC 31565 strain was treated with atmospheric and room temperature plasma (ARTP). Four stable mutants were obtained by ARTP, of which the A. pretiosum L-40 mutant produced 242.9 mg/L AP-3, representing a 22.5% increase compared to the original wild type strain. With seed medium optimization, AP-3 production of mutant L-40 reached 307.8 mg/L; qRT-PCR analysis revealed that AP-3 biosynthesis-related gene expression was significantly up-regulated under optimized conditions. To further improve the AP-3 production, genome shuffling (GS) technology was used on the four A. pretiosum mutants by ARTP. After three rounds of GS combined with high-throughput screening, the genetically stable recombinant strain G3-96 was obtained. The production of AP-3 in the G3-96 strain was 410.1 mg/L in shake flask cultures, which was 44.5% higher than the L-40 production from the parental strain, and AP-3 was increased by 93.8% compared to the wild-type A. pretiosum. These results suggest that the combination of mutagenesis, seed medium optimization, and GS technology can effectively improve the AP-3 production capacity of A. pretiosum and provide an enabling methodology for AP-3 industrial production.


Assuntos
Actinobacteria/crescimento & desenvolvimento , Proteínas de Bactérias/genética , Maitansina/análogos & derivados , Plasma/fisiologia , Actinobacteria/genética , Actinobacteria/metabolismo , Proteínas de Bactérias/metabolismo , Técnicas de Cultura Celular por Lotes , Embaralhamento de DNA , Fermentação , Maitansina/biossíntese , Engenharia Metabólica , Mutagênese
3.
Rev. cuba. invest. bioméd ; 40(1): e893, ene.-mar. 2021.
Artigo em Espanhol | LILACS, CUMED | ID: biblio-1289450

RESUMO

Introducción: El plasma de convalecientes es una inmunoterapia pasiva que se ha usado para el tratamiento y prevención de muchas enfermedades infecciosas por más de un siglo. Dada la falta de tratamiento específico para el nuevo coronavirus SARS-CoV-2, el plasma de convalecientes es una alternativa terapéutica potencial contra la COVID-19. Objetivo: Realizar una revisión del empleo del plasma de convalecientes como alternativa terapéutica a la COVID-19. Desarrollo: Se empleó la estrategia de búsqueda del tema; consultando las bases de datos Pubmed, SciELO, Lilacs, Cochrane Library y Web of Science. El plasma de convalecientes ha mostrado efectividad en el tratamiento de varias enfermedades virales. Así, la evidencia sobre su uso en los pacientes con COVID-19 es escasa, aunque se han obtenido resultados alentadores, pero no concluyentes por falta de un número mayor ensayos clínicos. Al mismo tiempo, Cuba incluye en sus protocolos de actuación contra la COVID-19 este tratamiento. Conclusiones: Esta alternativa resulta una herramienta inmunoterapéutica en los pacientes con la COVID-19, ya que mejora el estado clínico y disminuir la tasa de letalidad. Sin embargo, se necesitan más ensayos clínicos controlados y aleatorizados que afirmen su efectividad y seguridad(AU)


Introduction: Convalescent plasma is a form of passive immunotherapy which has been used for the treatment and prevention of many infectious diseases for more than one century. Given the absence of a specific treatment for the novel coronavirus SARS-CoV-2, convalescent plasma is a potential therapeutic alternative against COVID-19. Objective: Carry out a review about the use of convalescent plasma as a therapeutic alternative against COVID-19. Discussion: A search was conducted about the topic in the databases Pubmed, SciELO, Lilacs, Cochrane Library and Web of Science. Convalescent plasma has been shown to be effective in the treatment of several viral diseases. However, evidence of its use in COVID-19 patients is scant. Promising results have been obtained, though, but they are not conclusive due to the need of a larger number of clinical trials. In Cuba this treatment is included among the clinical management protocols for COVID-19. Conclusions: This alternative is an immunotherapeutic tool for the treatment of COVID-19 patients, since it improves their clinical status and reduces lethality rates. However, more controlled and randomized clinical trials are required confirming its effectiveness and safety(AU)


Assuntos
Humanos , Doenças Transmissíveis , Imunização Passiva , Coronavirus , Plasma/fisiologia
4.
Clin Pharmacol Ther ; 109(4): 1030-1033, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33547636

RESUMO

Boffito et al. recalled the critical importance to correctly interpret protein binding. Changes of lopinavir pharmacokinetics in coronavirus disease 2019 (COVID-19) are a perfect illustration. Indeed, several studies described that total lopinavir plasma concentrations were considerably higher in patients with severe COVID-19 than those reported in patients with HIV. These findings have led to a reduction of the dose of lopinavir in some patients, hypothesizing an inhibitory effect of inflammation on lopinavir metabolism. Unfortunately, changes in plasma protein binding were never investigated. We performed a retrospective cohort study. Data were collected from the medical records of patients hospitalized for COVID-19 treated with lopinavir/ritonavir in intensive care units or infectious disease departments of Toulouse University Hospital (France). Total and unbound concentrations of lopinavir, C reactive protein, albumin, and alpha-1-acid glycoprotein (AAG) levels were measured during routine care on the same samples. In patients with COVID-19, increased total lopinavir concentration is the result of an increased AAG-bound lopinavir concentration, whereas the unbound concentration remains constant, and insufficient to reduce the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) viral load. Although international guidelines have recently recommended against using lopinavir/ritonavir to treat severe COVID-19, the description of lopinavir pharmacokinetics changes in COVID-19 is a textbook case of the high risk of misinterpretation of a total drug exposure when changes in protein binding are not taken into consideration.


Assuntos
Antivirais/farmacocinética , Tratamento Farmacológico da COVID-19 , Lopinavir/farmacocinética , Plasma/fisiologia , Ligação Proteica/fisiologia , Idoso , Albuminas/metabolismo , Antivirais/uso terapêutico , Proteína C-Reativa/metabolismo , Feminino , Glicoproteínas/metabolismo , Humanos , Lopinavir/uso terapêutico , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , SARS-CoV-2 , Carga Viral
5.
Ther Apher Dial ; 25(3): 262-272, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32710797

RESUMO

Nowadays, therapeutic plasmapheresis (TP) is accepted as part of the treatment for specific groups of diseases. The availability of different methods, including double filtration and adsorption, increases selectivity for the removal of substances. However, the use of these techniques requires a thorough understanding of the characteristics and components of plasma. By considering pivotal papers from several databases, the aim of this narrative review is to describe the characteristics of plasma related to apheresis techniques. We have tried to cover the clinical implications including physiology, estimation of plasma volume, viscosity, and a description of its components including the size, volume of distribution, and half-lives of the different substances to be removed or maintained depending on the clinical situation and applied apheresis technique. Applying this knowledge will help us to choose the right method and dosage and improve the efficacy of the procedure by preventing or addressing any complications.


Assuntos
Remoção de Componentes Sanguíneos/métodos , Plasma/fisiologia , Plasmaferese/métodos , Humanos
6.
Shock ; 55(2): 189-197, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-32694397

RESUMO

BACKGROUND: Altered postinjury platelet behavior is recognized in the pathophysiology of trauma-induced coagulopathy (TIC), but the mechanisms remain largely undefined. Studies suggest that soluble factors released by injury may inhibit signaling pathways and induce structural changes in circulating platelets. Given this, we sought to examine the impact of treating healthy platelets with plasma from injured patients. We hypothesized that healthy platelets treated ex-vivo with plasma from injured patients with shock would impair platelet aggregation, while treatment with plasma from injured patients with significant injury burden, but without shock, would enhance platelet aggregation. METHODS: Plasma samples were isolated from injured patients (pretransfusion) and healthy donors at a Level I trauma center and stored at -80°C. Plasma samples from four separate patients in each of the following stratified clinical groups were used: mild injury/no shock (injury severity score [ISS] 2-15, base excess [BE]>-6), mild injury/with shock (ISS 2-15, BE≤-6), severe injury/no shock (ISS>25, BE>-6), severe injury/with shock (ISS>25, BE≤-6), minimal injury (ISS 0/1, BE>-6), and healthy. Platelets were isolated from three healthy adult males and were treated with plasma for 30 min. Aggregation was stimulated with a thrombin receptor agonist and measured via multiple-electrode platelet aggregometry. Data were normalized to HEPES Tyrode's (HT) buffer-only treated platelets. Associations of plasma treatment groups with platelet aggregation measures were tested with Mann-Whitney U tests. RESULTS: Platelets treated with plasma from patients with shock (regardless of degree of injury) had significantly impaired thrombin-stimulated aggregation compared with platelets treated with plasma from patients without shock (P = 0.002). Conversely, platelets treated with plasma from patients with severe injury, but without shock, had amplified thrombin-stimulated aggregation (P = 0.030). CONCLUSION: Shock-mediated soluble factors impair platelet aggregation, and tissue injury-mediated soluble factors amplify platelet aggregation. Future characterization of these soluble factors will support development of novel treatments of TIC.


Assuntos
Plaquetas/fisiologia , Plasma/fisiologia , Agregação Plaquetária/fisiologia , Ferimentos e Lesões/sangue , Adulto , Transtornos da Coagulação Sanguínea/sangue , Transtornos da Coagulação Sanguínea/etiologia , Humanos , Masculino , Pessoa de Meia-Idade , Choque/sangue , Choque/etiologia , Ferimentos e Lesões/complicações , Adulto Jovem
7.
Cornea ; 40(3): 299-302, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33290319

RESUMO

PURPOSE: To evaluate the efficacy of topical fresh frozen plasma (FFP) therapy on clinical symptoms, findings, and prognosis after anterior segment surgeries in patients with ligneous conjunctivitis (LC). METHODS: Retrospective case note review. RESULTS: Eleven eyes of 7 cases whose remission was not achieved after medical treatment such as topical corticosteroids, cyclosporine A, and heparin were included in the study. The median age of admission was 19 (1-49) years, median duration of FFP treatment was 48 (15-79) months, median follow-up period was 62 (16-114) months, and median age at symptom onset was 12 (4-252) months. Diagnosis was made according to clinical presentations, plasminogen activities, and response to treatment. Topical FFP that was prepared in our clinic was used in all cases. Surgeries (membrane excision, eyelid surgery, deep anterior lamellar keratoplasty, and cataract surgery) were performed after at least 1 month of FFP treatment. Prosthetic contact lens was applied to one eye. During the follow-up period, recurrences requiring membrane excision and side effects from topical FFP were not observed. CONCLUSIONS: LC is a rare membranous conjunctivitis that proceeds with remissions and recurrences. When it was shown that the etiology of LC is plasminogen deficiency, FFP became the only treatment option targeting the etiology. In this study, we observed that the topical FFP is an effective treatment method that prevents recurrence and ensures regression of membranes and safer anterior segment surgeries in LC.


Assuntos
Conjuntivite/tratamento farmacológico , Implante de Lente Intraocular , Facoemulsificação , Plasma/fisiologia , Plasminogênio/deficiência , Dermatopatias Genéticas/tratamento farmacológico , Administração Oftálmica , Adulto , Pré-Escolar , Conjuntivite/fisiopatologia , Feminino , Seguimentos , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Dermatopatias Genéticas/fisiopatologia , Adulto Jovem
8.
Sci Rep ; 10(1): 17973, 2020 10 21.
Artigo em Inglês | MEDLINE | ID: mdl-33087839

RESUMO

As a commonly used bone substitute material in the clinic, inorganic bovine bone has the characteristics of osteoconduction but not osteoinduction. This study aimed to treat inorganic bovine bone using nonthermal argon-oxygen plasma (NTAOP) to obtain greater bioreactivity for enhancing adhesion, proliferation and differentiation of mouse preosteoblast MC3T3-E1 cells. In this study, inorganic bovine bone was activated by NTAOP, and the surface characteristics were analyzed. MC3T3-E1 cells were then seeded onto the surface of inorganic bovine bone. Cell morphology, proliferation and osteogenic differentiation were examined. There was no obvious change in the surface morphology of specimens between the two groups. Regarding the elemental composition of the material, the amount of surface carbon was reduced, whereas oxygen, phosphorus and calcium levels were increased in the NTAOP group. Further studies showed that the NTAOP groups performed better than their untreated counterparts in terms of supporting cell proliferation and differentiation. Inorganic bovine bone treated with NTAOP can promote preosteoblast adhesion, proliferation and differentiation.


Assuntos
Argônio/farmacologia , Osso e Ossos/citologia , Osso e Ossos/metabolismo , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Osteoblastos/citologia , Osteoblastos/fisiologia , Osteogênese/fisiologia , Oxigênio/farmacologia , Plasma/fisiologia , Animais , Cálcio/metabolismo , Carbono/metabolismo , Bovinos , Adesão Celular , Camundongos , Osteoblastos/metabolismo , Oxigênio/metabolismo , Fósforo/metabolismo
9.
Curr Opin Allergy Clin Immunol ; 20(6): 557-564, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33044340

RESUMO

PURPOSE OF REVIEW: This review describes how plasma is sourced for fractionation into plasma-derived medicinal products (PDMPs), such as immunoglobulin (Ig) together with differences between plasma from whole blood (recovered plasma) and from plasmapheresis (source plasma) in terms of global plasma supply. Specific areas of growth in immunoglobulin use are identified alongside novel therapies, which may reduce demand for some immunoglobulin indications. RECENT FINDINGS: There has been a 6--8% annual growth in immunoglobulin use. Secondary immunodeficiency alongside improved recognition and diagnosis primary immunodeficiency disorders are drivers whereas the novel neonatal Fc receptor inhibitors (FcRni) may reduce demand for some immunomodulatory indications. SUMMARY: There is a significant geographical imbalance in global supply of plasma with 65% collected in the United States. This results in a dependency of other countries on United States supply and argues for both more plasma supply and greater regionally balanced plasma collection. In addition, progress towards a transparent, regulated and well tolerated framework for the coexistence of unpaid and compensated plasma donations is needed as unpaid donation will not be sufficient. These discussions should be informed by the needs of patients for this life-saving therapy, the care of donors and the safety of plasma and PDMPs.


Assuntos
Antígenos de Histocompatibilidade Classe I/uso terapêutico , Imunoglobulinas/uso terapêutico , Síndromes de Imunodeficiência/terapia , Plasma/fisiologia , Receptores Fc/uso terapêutico , Doadores de Sangue , Humanos , Imunomodulação
10.
J Struct Biol ; 212(2): 107615, 2020 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-32927057

RESUMO

Visualizing bone mineralization and collagen fibril organization at intermediate scales between the nanometer and the hundreds of microns range, is still an important challenge. Similarly, visualizing cellular components which locally affect the tissue structure requires a precision of a few tens of nanometers at maximum while spanning several tens of micrometers. In the last decade, gallium focused ion beam (FIB) equipped with a scanning electron microscope (SEM) proved to be an extremely valuable structural tool to meet those ends. In this study, we assess the capability of a recent plasma FIB-SEM technology which provides a potential increase in measurement speed over gallium FIB-SEM, thus paving the way to larger volume analysis. Nanometer-scale layers of demineralized and mineralized unstained human femoral lamellar bone were sequentially sectioned over volumes of 6-16,000 µm3. Analysis of mineralized tissue revealed prolate ellipsoidal mineral clusters measuring approximately 1.1 µm in length by 700 nm at their maximum diameter. Those features, suggested by others in high resolution studies, appear here as a ubiquitous motif in mineralized lamellar bone over thousands of microns cubed, suggesting a heterogeneous and yet regular pattern of mineral deposition past the single collagen fibril level. This large scale view retained sufficient resolution to visualize the collagen fibrils while also partly visualizing the lacuno-canalicular network in three-dimensions. These findings are strong evidence for suitability of PFIB as a bone analysis tool and the need to revisit bone mineralization over multi-length scales with mineralized tissue.


Assuntos
Calcificação Fisiológica/fisiologia , Osso Cortical/fisiologia , Idoso , Calcinose/fisiopatologia , Matriz Extracelular/fisiologia , Fêmur/fisiologia , Humanos , Imageamento Tridimensional/métodos , Masculino , Microscopia Eletrônica de Varredura/métodos , Plasma/fisiologia
11.
Clin Lab ; 66(6)2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32538058

RESUMO

BACKGROUND: Applicability of thrombin generation tests to clinical routine has been sought for many years. The aim of this study was to compare thrombin generation measured in fresh platelet poor plasma (f-PPP) and frozen-thawed platelet poor plasma (ft-PPP) to prove the consistency of results. METHODS: In this prospective study, thrombin generation was measured in twenty-fold repetitions in 3.2% citrate PPP obtained from male healthy blood donors aged 19 - 39 years (n = 54 donations). The tests were performed with fresh PPP and repeated after storing the PPP at -60°C. In two subgroup analyses, the effect of higher and lower normal baseline platelet counts on the Calibrated Automated Thrombogram (CAT) assay and the influence of ABO blood groups on thrombin generation were analyzed. RESULTS: Referring to the parameters of thrombin generation most frequently used in studies, peak thrombin of f-PPP and ft-PPP agreed in about 50% of the samples. Endogenous thrombin potential (ETP) of f-PPP and ft-PPP agreed in nearly two-thirds of the samples. A slightly but significantly slower kinetic was found in the thrombin generation of ft-PPP compared with f-PPP. At least in f-PPP, ETP correlates with baseline platelet counts of the whole blood sample. Peak thrombin was significantly higher in non-O blood groups compared to O blood group. CONCLUSIONS: A low level of agreement between the results of f-PPP and ft-PPP is shown. In terms of practicability of sample collection using semi-automated thrombin-generation assays ft-PPP should be preferred over f-PPP. We therefore recommend using ft-PPP in clinical studies.


Assuntos
Antígenos de Grupos Sanguíneos , Preservação de Sangue/métodos , Coleta de Amostras Sanguíneas/métodos , Trombina , Adulto , Testes de Coagulação Sanguínea/métodos , Testes de Coagulação Sanguínea/normas , Criopreservação/métodos , Voluntários Saudáveis , Humanos , Masculino , Plasma/fisiologia , Contagem de Plaquetas/métodos , Trombina/análise , Trombina/metabolismo
12.
Aging (Albany NY) ; 12(10): 8790-8819, 2020 05 30.
Artigo em Inglês | MEDLINE | ID: mdl-32474458

RESUMO

Heterochronic blood sharing rejuvenates old tissues, and most of the studies on how this works focus on young plasma, its fractions, and a few youthful systemic candidates. However, it was not formally established that young blood is necessary for this multi-tissue rejuvenation. Here, using our recently developed small animal blood exchange process, we replaced half of the plasma in mice with saline containing 5% albumin (terming it a "neutral" age blood exchange, NBE) thus diluting the plasma factors and replenishing the albumin that would be diminished if only saline was used. Our data demonstrate that a single NBE suffices to meet or exceed the rejuvenative effects of enhancing muscle repair, reducing liver adiposity and fibrosis, and increasing hippocampal neurogenesis in old mice, all the key outcomes seen after blood heterochronicity. Comparative proteomic analysis on serum from NBE, and from a similar human clinical procedure of therapeutic plasma exchange (TPE), revealed a molecular re-setting of the systemic signaling milieu, interestingly, elevating the levels of some proteins, which broadly coordinate tissue maintenance and repair and promote immune responses. Moreover, a single TPE yielded functional blood rejuvenation, abrogating the typical old serum inhibition of progenitor cell proliferation. Ectopically added albumin does not seem to be the sole determinant of such rejuvenation, and levels of albumin do not decrease with age nor are increased by NBE/TPE. A model of action (supported by a large body of published data) is that significant dilution of autoregulatory proteins that crosstalk to multiple signaling pathways (with their own feedback loops) would, through changes in gene expression, have long-lasting molecular and functional effects that are consistent with our observations. This work improves our understanding of the systemic paradigms of multi-tissue rejuvenation and suggest a novel and immediate use of the FDA approved TPE for improving the health and resilience of older people.


Assuntos
Albuminas/farmacologia , Camadas Germinativas , Troca Plasmática , Plasma/fisiologia , Rejuvenescimento/fisiologia , Envelhecimento/efeitos dos fármacos , Envelhecimento/fisiologia , Animais , Células Cultivadas , Camadas Germinativas/citologia , Camadas Germinativas/fisiologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Solução Salina/farmacologia
13.
Life Sci ; 254: 117780, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32407844

RESUMO

AIMS: In vivo studies suggest a positive influence of fresh frozen plasma (FFP) on endothelial properties and vascular barrier function, leading to improved outcomes in animal sepsis models as well as in major abdominal surgery. However, those effects are incompletely described. It was our aim to evaluate in vitro effects of FFP on endothelial key functions and to identify underlying mechanisms. MATERIALS AND METHODS: Human pulmonary microvascular endothelial cells (HPMECs) were prestimulated with LPS, followed by incubation with FFP. Permeability for FITC-dextran was assessed, and intercellular gap formation was visualized. NF-κB nuclear translocation and expression of pro-inflammatory, pro-adhesion, and leakage-related genes were evaluated, and monocyte adhesion to ECs was assessed. Intracellular cAMP levels as well as phosphorylation of functional proteins were analyzed. In patients undergoing major abdominal surgery, Syndecan-1 serum levels were assessed prior to and following FFP transfusion. KEY FINDINGS: Post-incubation of HPMVECs with FFP increased intracellular cAMP levels that had been decreased by preceding LPS stimulation. On one hand, this reduced endotoxin-mediated upregulation of IL-8, ICAM-1, VCAM-1, VEGF, and ANG-2. Impaired phosphorylation of functional proteins was restored, and intercellular cohesion and barrier function were rescued. On the other hand, NF-κB nuclear translocation as well as monocyte adhesion was markedly increased by the combination of LPS and FFP. Syndecan-1 serum levels were lower in surgery patients that were transfused with FFP compared to those that were not. SIGNIFICANCE: Our data provide evidence for a differential modulation of crucial endothelial properties by FFP, potentially mediated by elevation of intracellular cAMP levels.


Assuntos
Células Endoteliais/fisiologia , Endotélio Vascular/metabolismo , Plasma/fisiologia , Idoso , Adesão Celular/fisiologia , Permeabilidade da Membrana Celular/fisiologia , Células Cultivadas , AMP Cíclico/metabolismo , Dextranos/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Junções Comunicantes/fisiologia , Humanos , Lipopolissacarídeos , Pessoa de Meia-Idade , Monócitos/fisiologia , NF-kappa B/metabolismo , Fosforilação , Sindecana-1/sangue
14.
Wilderness Environ Med ; 31(2): 131-143, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32336579

RESUMO

INTRODUCTION: Hump-nosed pit vipers (Hypnale spp) cause the highest number of venomous snakebites in Sri Lanka. Bites commonly cause local envenoming leading to local pain, swelling, and necrosis of the site of the bite. Acute kidney injury is the most common systemic manifestation, and some patients develop venom-induced consumption coagulopathy (VICC). Genus Hypnale comprises 3 species. Of them, H hypnale is found in Sri Lanka and the Western Ghats region of India. The other 2 (H nepa and H zara) are endemic species in Sri Lanka. METHODS: This study included 500 patients with hump-nosed viper bites studied prospectively over 4.5 y starting June 2014. All patients were assessed and the data were collected by the principal investigator (primary data). A subgroup of patients who developed VICC is described. There were 2 groups, including proven (patients with the specimen of the snake) and probable (specimen of snake not available) bites. RESULTS: Thirty (n=500; 6%) patients developed VICC; of them, 17 (3%) were proven cases, and 13 (2%) were probable cases. In both groups, 24 (80%) recovered, 2 (7%) progressed to chronic kidney disease, 1 (3%) died of severe hemostatic dysfunction, and 3 (10%) were lost to follow-up. Systemic bleeding was observed in 16 patients (53%), including hematuria (microscopic and gross) in 8 (27%) and venipuncture bleeding in 5 (17%). Eleven (37%) developed local bleeding at the site of the bite. Fresh frozen plasma was administered to 20 patients (67%), among whom only 11 (55%) experienced early correction of VICC. In both groups, 15 (50%) developed acute kidney injury, and 2 (7%) progressed to chronic kidney disease. Microangiopathic hemolysis was observed in 18 patients (60%) and thrombocytopenia in 16 (53%). Thrombotic microangiopathy was detected in 13 patients (43%), of whom 10 (33%) developed hemolytic uremic syndrome and 2 (7%) had thrombotic thrombocytopenic purpura. Of patients with VICC in the proven group, 94% (n=16) was caused by H hypnale and 1 (6%) was caused by H zara. In the proven group, median international normalized ratio was 3.7 (interquartile range 1.6-5.0); in the probable group, it was 5.0 (interquartile range 2.1-5.4). CONCLUSIONS: We found that 6% of patients develop hemostatic dysfunction after hump-nosed viper bites. However, which patients will develop coagulopathy or die of envenoming is unpredictable. Reliable and accessible treatments are unmet essential needs because antivenoms for these bites are currently not available in the country. Therapy with fresh frozen plasma has doubtful efficacy in early correction of VICC and needs further evaluation.


Assuntos
Coagulação Intravascular Disseminada/terapia , Plasma/fisiologia , Mordeduras de Serpentes/complicações , Venenos de Víboras/efeitos adversos , Viperidae , Adulto , Idoso , Animais , Coagulação Intravascular Disseminada/induzido quimicamente , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Mordeduras de Serpentes/terapia , Sri Lanka
15.
Transfusion ; 60(5): 1004-1014, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32167176

RESUMO

BACKGROUND: Most chimeric antigen receptor T (CAR-T) cells and other adoptive T-cell therapies (ACTs) are currently manufactured by ex vivo expansion of patient lymphocytes in culture media supplemented with human plasma from group AB donors. As lymphocytes do not express A or B antigens, the isoagglutinins of non-AB plasmas are unlikely to cause deleterious effects on lymphocytes in culture. STUDY DESIGN AND METHODS: Seeding cultures with peripheral blood mononuclear cell (PBMNC) concentrates from group A1 donors and using a CAR-T culture protocol, parallel cultures were performed, each with unique donor plasmas as media supplements (including group O plasmas with high-titer anti-A and group AB plasmas as control). An additional variable, a 3% group A1 red blood cell (RBC) spike, was added to simulate a RBC-contaminated PBMNC collection. Cultures were monitored by cell count, viability, flow cytometric phenotype, gene expression analysis, and supernatant chemokine analysis. RESULTS: There was no difference in lymphocyte expansion or phenotype when cultured with AB plasma or O plasma with high-titer anti-A. Compared to controls, the presence of contaminating RBCs in lymphocyte culture led to poor lymphocyte expansion and a less desirable phenotype-irrespective of the isoagglutinin titer of the plasma supplement used. CONCLUSIONS: This study suggests that ABO incompatible plasma may be used as a media supplement when culturing cell types that do not express ABO antigens-such as lymphocytes for CAR-T or other ACT. The presence of contaminating RBCs in culture was disadvantageous independent of isoagglutinin titer.


Assuntos
Sistema ABO de Grupos Sanguíneos/sangue , Meios de Cultura/química , Imunoterapia Adotiva , Plasma/fisiologia , Cultura Primária de Células/métodos , Linfócitos T/citologia , Células Cultivadas , Citometria de Fluxo , Antígenos HLA-A/sangue , Antígenos HLA-A/imunologia , Humanos , Imunoterapia Adotiva/métodos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/transplante , Ativação Linfocitária , Plasma/química , Cultura Primária de Células/normas , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/metabolismo , Receptores de Antígenos Quiméricos/genética , Receptores de Antígenos Quiméricos/metabolismo , Linfócitos T/metabolismo , Linfócitos T/transplante , Doadores de Tecidos
16.
Ir J Med Sci ; 189(2): 603-609, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31735988

RESUMO

BACKGROUND: Obstructive sleep apnea-hypopnea syndrome (OSAHS) is a common sleep-related respiratory disease. Despite reports of low-temperature plasma radiofrequency ablation of tonsils and adenoids for the treatment of OSAHS, the effects on lung function and quality of life are unclear. AIMS: This study aims to explore the effects of low-temperature plasma treatment on pulmonary function in children with OSAHS. METHODS: A total of 110 children with OSAHS were included in this prospective study. Low-temperature plasma radiofrequency treatment and routine surgical treatment were performed in group A and group B, respectively. Maximal voluntary ventilation (MVV), forced vital capacity (FVC), and total lung capacity (TLC) were measured. OSA-18 survey was used to evaluate the quality of life 1 year after operation. RESULTS: Group A had significantly higher effective treatment rate (P < 0.05) and lower operative period, bleeding volume, visual analog score, pseudomembrane detachment time, and time required to feed after operation than group B (all P < 0.001). There were no significant difference in the incidence of postoperative complications between the groups and MVV, FVC, and TLC between the groups before and after operation (all P > 0.05), and the above indexes in both groups were significantly increased after operation (all P < 0.05). OSA-18 scores in group A were significantly lower than group B after operation (P < 0.001). CONCLUSIONS: Low-temperature plasma technique performed in OSAHS children has a good clinical effect, which can reduce the operation time, intraoperative blood loss, postoperative pain, pseudomembrane detachment time, hospitalization time, and improve pulmonary function and quality of life.


Assuntos
Plasma/fisiologia , Qualidade de Vida/psicologia , Testes de Função Respiratória/métodos , Apneia Obstrutiva do Sono/complicações , Criança , Feminino , Humanos , Masculino , Estudos Prospectivos , Síndrome , Temperatura , Resultado do Tratamento
17.
Int J Mol Sci ; 20(20)2019 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-31640211

RESUMO

Recent studies have shown that plasma can efficiently inactivate microbial pathogens such as bacteria, fungi, and viruses in addition to degrading toxins. Moreover, this technology is effective at inactivating pathogens on the surface of medical and dental devices, as well as agricultural products. The current practical applications of plasma technology range from sterilizing therapeutic medical devices to improving crop yields, as well as the area of food preservation. This review introduces recent advances and future perspectives in plasma technology, especially in applications related to disinfection and sterilization. We also introduce the latest studies, mainly focusing on the potential applications of plasma technology for the inactivation of microorganisms and the degradation of toxins.


Assuntos
Desinfecção/métodos , Plasma/fisiologia , Esterilização/métodos , Agricultura , Equipamentos e Provisões/microbiologia , Conservação de Alimentos , Humanos
18.
Transfusion ; 59(11): 3492-3500, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31568583

RESUMO

BACKGROUND: Platelet-rich plasma (PRP) is frequently used in the treatment of acute and chronic wounds. One of the major problems concerning the use of PRP is the absence of a well-characterized and standardized product, which leads to a high variety in study outcomes. Therefore, more studies on the composition and standardization of PRP in wound healing are needed. STUDY DESIGN AND METHODS: Platelet concentrates derived from healthy blood donors were made in plasma (PC-plasma) or platelet additive solution (PC-PAS). The effects of PC-plasma, PC-PAS, and plasma were then tested on proliferation, differentiation, and migration of fibroblasts, as well as sprouting of endothelial cells in fibrin gels and chemotaxis of white blood cells (WBCs). RESULTS: PC-plasma stimulates the migration and proliferation of human dermal fibroblasts more than plasma or platelets alone. Furthermore, platelet factors decrease the expression of α-smooth muscle actin in dermal fibroblast cultures. PC-plasma also stimulates sprouting of endothelial cells. Finally, PC-plasma also acts as a strong chemoattractant for WBCs. CONCLUSIONS: Allogeneic PC-plasma has beneficial effects on various aspects of wound healing in vitro and is superior to plasma or platelets alone. PC-plasma is an attractive candidate for further in vivo evaluation.


Assuntos
Plaquetas/fisiologia , Quimiotaxia de Leucócito , Fibroblastos/fisiologia , Neovascularização Fisiológica , Plasma/fisiologia , Plasma Rico em Plaquetas/fisiologia , Cicatrização/fisiologia , Actinas/sangue , Movimento Celular , Proliferação de Células , Células Cultivadas , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/sangue
20.
Cells ; 8(9)2019 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-31500151

RESUMO

Extracellular vesicles (EVs) isolated from biological samples are a promising material for use in medicine and technology. However, the assessment methods that would yield repeatable concentrations, sizes and compositions of the harvested material are missing. A plausible model for the description of EV isolates has not been developed. Furthermore, the identity and genesis of EVs are still obscure and the relevant parameters have not yet been identified. The purpose of this work is to better understand the mechanisms taking place during harvesting of EVs, in particular the role of viscosity of EV suspension. The EVs were harvested from blood plasma by repeated centrifugation and washing of samples. Their size and shape were assessed by using a combination of static and dynamic light scattering. The average shape parameter of the assessed particles was found to be ρ ~ 1 (0.94-1.1 in exosome standards and 0.7-1.2 in blood plasma and EV isolates), pertaining to spherical shells (spherical vesicles). This study has estimated the value of the viscosity coefficient of the medium in blood plasma to be 1.2 mPa/s. It can be concluded that light scattering could be a plausible method for the assessment of EVs upon considering that EVs are a dynamic material with a transient identity.


Assuntos
Viscosidade Sanguínea/fisiologia , Vesículas Extracelulares/fisiologia , Manejo de Espécimes/métodos , Adulto , Centrifugação/métodos , Difusão Dinâmica da Luz/métodos , Exossomos/fisiologia , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Plasma/fisiologia , Viscosidade
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