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1.
J Clin Lab Anal ; 35(10): e23955, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34424578

RESUMO

BACKGROUND: Stored red blood cells (RBCs) may undergo oxidative stress over time, with functional changes affecting oxygen delivery. Central to these changes are oxidation-reduction (redox) reactions and redox potential (RP) that must be maintained for cell function. RP imbalance can lead to oxidative stress that may contribute to storage lesions. This study's purpose was to identify changes in RP over time in banked RBCs, and among RBCs of similar age. METHODS: Multiple random RBC segments from RBC units were tested (n = 32), ranging in age from 5 to 40 days, at 5-day intervals. RP was recorded by measuring open circuit potential of RBCs using nanoporous gold electrodes with Ag/AgCl reference. RP measures were also performed on peripheral venous blood from 10 healthy volunteers. RP measures were compared between RBC groups, and with volunteer blood. RESULTS: Stored RBCs show time-dependent RP increases. There were significant differences in Day 5 RP compared to all other groups (p ≤ 0.005), Day 10-15 vs. ages ≥ Day 20 (p ≤ 0.025), Day 20-25 vs. Day 40 (p = 0.039), and all groups compared to healthy volunteers. RP became more positive over time suggesting ongoing oxidation as RBCs age; however, storage time alone was not predictive of RP measured in a particular unit/segment. CONCLUSIONS: There are significant differences in RP between freshly stored RBCs and all others, with RP becoming more positive over time. However, storage time alone does not predict RP, indicating RP screening may be an important measure of RBC oxidative stress and serve as an RBC quality marker.


Assuntos
Preservação de Sangue , Eritrócitos/fisiologia , Estresse Oxidativo/fisiologia , Bancos de Sangue , Transfusão de Eritrócitos , Humanos , Oxirredução
2.
Micromachines (Basel) ; 11(8)2020 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-32707873

RESUMO

The goal of this study is the preparation of safer coated microneedles so that tips remaining after the initial use are less likely to be reinserted on a second use. Twelve groups of uncoated microneedles (u-MNs) were prepared from the combination of three different aspect ratios (height to base width) and four kinds of polymer (polyethylene (PE), polypropylene (PP), nylon and polylactic acid (PLA)). After coating the u-MNs with polyvinyl alcohol formulation to make coated MNs (c-MNs), the force displacement of the u-MNs and the c-MNs was measured. The aspect ratio was reduced from 2.2, 2.5 and 3.0 with u-MNs to 1.3, 1.4 and 1.6 with c-MNs, respectively, after the coating formulation was applied to the MNs. All PLA MNs had a puncture performance of more than 95%. However, the puncture performance of u-MNs made of PE and of PP with a 3.0 aspect ratio was only 8% and 53%, respectively, whereas the rates of c-MNs made of PE and of PP were 82% and 95%, respectively. In animal experiments with PP MNs with a 3.0 aspect ratio, the 59% rate of puncture performance with u-MNs increased to above 96% with c-MNs and fell to 13% for r-MNs. Safe c-MNs can overcome the disadvantages of standard c-MNs by reducing the probable contamination of remaining tips after use. Safe c-MNs have advantages over standard c-MNs in terms of humidity resistance, reasonable cost, sterilization process and short processing time through the separate process of u-MN preparation and simple dip-coating.

3.
J Control Release ; 286: 460-466, 2018 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-30102940

RESUMO

In this study, we present transcutaneous influenza vaccination using a novel tip-separable microneedle system called insertion-responsive microneedles (IRMNs). IRMNs are composed of dissolvable hyaluronic acid (HA) tips and biocompatible polycaprolactone (PCL) bases, the tip of which is instantly separated from the base during microneedle insertion and retraction. Vaccine antigens derived from canine influenza virus (A/canine/VC378/2012; H3N2) were successfully coated on HA tips by rapidly freezing the tips prior to coating. An ex vivo porcine skin insertion test showed that IRMNs were capable of penetrating the skin without tip breakage and releasing the coated materials within the skin. The thermal stability of the vaccine as determined by hemagglutination assay revealed that the coated vaccine partially maintained its activity when stored at 50 °C for 3 weeks, whereas the liquid form completely lost the activity. Immunization in guinea pigs showed that hemagglutination inhibition (HI) antibodies induced by IRMNs were two times higher than those induced by intramuscular (IM) injections. When challenged with influenza A/canine/Korea/01/2007 (H3N2) wild-type virus 2 weeks after the second vaccination, viral shedding was completely eliminated at 8 days post infection in both IRMNs and IM injection groups. Our results suggest that IRMNs have great potential for rapid and convenient vaccination, which will be particularly attractive for animal vaccinations.


Assuntos
Doenças do Cão/prevenção & controle , Vacinas contra Influenza/administração & dosagem , Infecções por Orthomyxoviridae/prevenção & controle , Vacinação/instrumentação , Animais , Linhagem Celular , Doenças do Cão/imunologia , Cães , Sistemas de Liberação de Medicamentos/economia , Sistemas de Liberação de Medicamentos/instrumentação , Desenho de Equipamento , Feminino , Cobaias , Vírus da Influenza A Subtipo H3N2/imunologia , Vacinas contra Influenza/imunologia , Vacinas contra Influenza/uso terapêutico , Injeções Intradérmicas , Microinjeções/economia , Microinjeções/instrumentação , Agulhas , Infecções por Orthomyxoviridae/imunologia , Suínos , Fatores de Tempo , Vacinação/economia
4.
Shock ; 49(3): 345-351, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-28658006

RESUMO

INTRODUCTION: Oxidation-reduction reactions involve electron exchanges that require optimal balance for proper cell function. This balance is measured via redox potential and reflects oxidative stress. Despite the critical role of oxidative stress in critical illness and injury, little is known regarding redox potential. We hypothesize redox potential measurements will correlate with accumulation of O2 debt produced by hemorrhage over time. METHODS: Ten swine were studied using a polytrauma hemorrhagic shock model. Whole blood and plasma redox potential measures were obtained at defined stages of O2 debt (20 mL/kg, 40 mL/kg, 60 mL/kg, 80 mL/kg), and through resuscitation. Redox potential was determined by measuring open circuit potential using novel gold nanoporous electrodes with Ag/AgCl reference. RESULTS: Whole blood redox potential showed negative change as O2 debt accumulated, exhibiting positive response during resuscitation, and correlated with O2 debt across all animals (P < 0.001). Redox potential changes throughout O2 debt accrual were significant compared with baseline (P≤0.05), and at end resuscitation compared with O2 debt 60 mL/kg (P = 0.05) and 80 mL/kg (P = 0.02). Whole blood redox potential measures also correlated with oxygen extraction ratio, ScvO2, and lactic acid, appearing very sensitive to acute changes. Plasma redox potential showed no correlation with O2 debt. CONCLUSIONS: Whole blood redox potential demonstrates significant correlation to O2 debt at all stages in this model. These results set the stage for further study of redox potential as a direct measure of oxidative stress and potential clinical tool. Given redox potential plasma performance, these measures should be made in whole blood versus plasma.


Assuntos
Traumatismo Múltiplo/sangue , Oxigênio/sangue , Choque Hemorrágico/sangue , Animais , Biomarcadores/sangue , Modelos Animais de Doenças , Humanos , Traumatismo Múltiplo/patologia , Oxirredução , Choque Hemorrágico/patologia , Suínos
5.
RSC Adv ; 8(32): 17786-17796, 2018 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-35542069

RESUMO

We have developed an insertion-responsive microneedle (IRMN) system that enables prompt drug delivery through the skin without attaching a skin patch. This system consists of square pyramidal hyaluronic acid (HA) microneedle tips and polycaprolactone (PCL) base arrays. During skin insertion, HA tips can be immediately separated from PCL base arrays due to the relatively weak adhesion strength between HA and PCL. Two base designs using truncated square pyramid stands, one without a wall (no-walled stand, NWS) and another with a wall on one side of the stand (single-walled stand, SWS), were prepared to study the effect of base geometry on the mechanical behavior of IRMNs. Ex vivo skin insertion tests showed successful separation of the tips from the base array upon insertion, regardless of the presence of a wall on the stand. However, only IRMNs-SWS were deeply embedded within the skin. Mechanical testing results demonstrated that the presence of a wall on the base enhanced the mechanical stability of the IRMNs. The wall also provided adequate adhesion between the tips and base, preventing the tips from breaking during insertion, while allowing the needle tip to separate upon removal. Histological examination confirmed that the tips were successfully separated from the base, embedded in the skin, and released fluorescent dyes within the skin. Our results suggest that the IRMN system is promising for the rapid and accurate delivery of various molecules through the skin, while improving user convenience by eliminating the need to attach microneedles to the skin.

6.
J Control Release ; 207: 1-6, 2015 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-25828366

RESUMO

A hollow microneedle (HM) was prepared to deliver a phenylephrine (PE) solution into the anal sphincter muscle as a method for treating fecal incontinence. The goal of this study was the local targeted delivery of PE into the sphincter muscle through the perianal skin with minimal pain using hollow microneedles, resulting in the increase of resting anal sphincter pressure. PE was administered on the left and the right sides of the anus of a rat through the perianal skin using 1.5mm long HM. An in vivo imaging system study was conducted after injection of Rhodamine B, and a histological study was performed after injection of gentian violet. The resting anal sphincter pressure in response to various drug doses was measured by using an air-charged catheter. Anal pressure change produced by HM administration was compared with change produced by intravenous injection (IV), subcutaneous (SC) injection and intramuscular (IM) injection. The change in mean blood pressure produced by HM administration as a function of PE dose was compared with change produced by PBS injection. A pharmacokinetic study of the new HM administration method was performed. A model drug solution was localized in the muscle layer under the perianal skin at the injection site and then diffused out over time. HM administration of PE induced significant contraction of internal anal sphincter pressure over 12h after injection, and the maximum anal pressure was obtained between 5 and 6h. Compared to IV, SC and IM treatments, HM treatment produced greater anal pressure. There was no increase in blood pressure after HM administration of PE within the range of predetermined concentration. Administration of 800µg/kg of PE using HM produced 0.81±0.38h of tmax. Our study suggests that HM administration enables local delivery of a therapeutic dose of PE to the anal sphincter muscle layer with less pain. This new treatment has great potential as a clinical application because of the ease of the procedure, minimal pain, and dose-dependent response.


Assuntos
Agonistas de Receptores Adrenérgicos alfa 1/administração & dosagem , Canal Anal/efeitos dos fármacos , Sistemas de Liberação de Medicamentos/instrumentação , Incontinência Fecal/tratamento farmacológico , Fenilefrina/administração & dosagem , Agonistas de Receptores Adrenérgicos alfa 1/farmacocinética , Agonistas de Receptores Adrenérgicos alfa 1/toxicidade , Canal Anal/fisiopatologia , Animais , Difusão , Relação Dose-Resposta a Droga , Incontinência Fecal/fisiopatologia , Feminino , Injeções Intramusculares , Injeções Intravenosas , Injeções Subcutâneas , Miniaturização , Agulhas , Fenilefrina/farmacocinética , Fenilefrina/toxicidade , Pressão , Ratos Sprague-Dawley , Distribuição Tecidual
7.
Yonsei Med J ; 47(6): 862-9, 2006 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-17191317

RESUMO

Our experiments aimed to clarify the mechanism by which host cell apoptosis is inhibited by infection with the intracellular protozoan parasite, Toxoplasma gondii (T. gondii). Mouse spleen cells were cultured in 6-well plates with RPMI 1640/ 10% FBS at 37?, in a 5% CO2 atmosphere. Apoptosis of spleen cells was induced by actinomycin-D (AD) treatment for 1 h prior to infection with T. gondii. A variety of assays were used to assess the progression of apoptosis: DNA size analysis on agarose gel electrophoresis, flow cytometry with annexin V/PI staining, and analysis of expression levels of Bcl-2 family and NF-kappaB mRNA and proteins by RT-PCR, Western blotting, and EMSA. Additionally, transmission electron microscopy (TEM) was performed to observe changes in cell morphology. Fragmentation of DNA was inhibited in spleen cells treated with AD and T. gondii 5 h and 18 h post infection, respectively, and flow cytometry studies showed a decreased apoptotic rates in AD and T. gondii treated spleen cells. We observed decreased expression of Bax mRNA and protein, while levels of Bcl-2 mRNA remained constant in spleen cells treated with AD and T. gondii. Caspase 3 and PARP were inactivated in cells treated with AD and T. gondii, and increased levels of cleaved caspase 8 were also observed. Analysis of EMSA and Western blot data suggests that activation of transcription factor NF-kappaB may be involved in the blockade of apoptosis by T. gondii. TEM analysis showed nuclear fragmentation and chromatin condensation occurring in spleen cells treated with AD; however, such apoptosis- associated morphological changes were not observed in cells treated with both AD and T. gondii tachyzoites. Together, these data show that T. gondii infection inhibits AD induced apoptosis via caspase inactivation and NF-kappaB activation in mouse spleen cells.


Assuntos
Apoptose/fisiologia , Inibidores de Caspase , NF-kappa B/metabolismo , Toxoplasma/fisiologia , Animais , Células Cultivadas , Fragmentação do DNA , Citometria de Fluxo , Regulação da Expressão Gênica , Camundongos , Inibidores de Poli(ADP-Ribose) Polimerases , RNA Mensageiro/metabolismo , Proteína X Associada a bcl-2/metabolismo
8.
Fertil Steril ; 85(5): 1512-5, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16566932

RESUMO

Y-chromosome microdeletion in male fetuses conceived by intracytoplasmic sperm injection (ICSI) was screened by polymerase chain reaction for sequence-tagged sites in azoospermia factor (AZF)-b and AZFc regions. Treatment with ICSI may lead to vertical transmission, expansion, and de novo Y-chromosome microdeletion in male fetuses.


Assuntos
Deleção Cromossômica , Transtornos Cromossômicos/genética , Cromossomos Humanos Y/genética , Transmissão Vertical de Doenças Infecciosas , Infertilidade Masculina/genética , Infertilidade Masculina/terapia , Injeções de Esperma Intracitoplásmicas , Humanos , Incidência , Masculino , Mosaicismo , Fatores Sexuais
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