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1.
Int J Biol Macromol ; 250: 126107, 2023 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-37536417

RESUMEN

Amylopectin clusters (APCs) are produced by cyclodextrin glucanotransferase (EC 2.4.1.19). Their solubility rate in aqueous solution was found to be 16.7 %. The weight-average molecular weight of APCs is ∼105 Da, as determined by multiangle laser light scattering analysis. Side chain length analysis indicated that the relative proportions of side chains with a degree of polymerization in the ranges of 2-8 and 25-50 decreased and increased, respectively, during preparation of APCs. In the exercise experiment, the blood glucose level of rats was higher in the APC-treated group than in the groups treated with commercial carbohydrate supplement (CCD) and glucose. In the forced swimming test, the swimming time in the APC and CCD groups increased by 22.6 % and 31.1 %, respectively, compared with the glucose administration group. The insulin levels were also similar between the APC and CCD groups. However, the glycogen levels in the liver and muscles of mice were significantly higher in the APC group than control group. These results suggest that APCs could potentially enhance endurance when added to sports drinks.

2.
Carbohydr Polym ; 310: 120722, 2023 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-36925261

RESUMEN

To improve the applicability of quercetin (QCT), we produced a QCT and cycloamylose (CA-QCT) inclusion complex based on the cyclization activity of cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19). The encapsulated QCT was purified using recycling preparative high-performance liquid chromatography, and its formation was analyzed using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The water solubility of CA-QCT was 55,000-fold higher than that of QCT. CA-QCT had 97 % stability for one week at pH 8 in a 4 °C water bath. According to a 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity assay, CA-QCT activity in aqueous solution was 24 times higher than that of an equal amount of QCT in aqueous solution. In an anti-inflammatory assay using lipopolysaccharide-induced RAW264.7 macrophages, CA-QCT in aqueous solution decreased nitric oxide production in a similar manner to QCT in dimethyl sulfoxide (DMSO). Additionally, even under aqueous conditions, CA-QCT more effectively inhibited the production of inflammatory mediators, such as interleukin-1ß, interleukin-6, and cyclooxygenase, compared with QCT dissolved in DMSO.


Asunto(s)
Dimetilsulfóxido , Quercetina , Quercetina/farmacología , Quercetina/química , Antiinflamatorios/farmacología , Oligosacáridos
3.
Foods ; 12(3)2023 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-36766169

RESUMEN

In the rice processing industry, wastewater is an inevitable by-product of rice washing. To increase the utilization of washed rice water (WRW), seven types of fermented washed rice water (FWRW) were prepared using lactic acid bacteria (LAB) and carbohydrate hydrolase. The total concentration of small maltooligosaccharides (MOSs) in the amyloglucosidase (AMG) treatment groups was about ten times higher than in the untreated groups. After 6 h of fermentation, six of the seven FWRW samples reached a pH of 4 due to the increased concentration of organic acids and could, therefore, be used as food acidity regulators. To confirm the applicability of FWRW, the traditional Korean rice cake garaetteok was prepared with FWRW and stored at 4 °C for 5 days. A texture profile analysis (TPA) revealed that the hardness of garaetteok treated with FWRW was significantly lower than that of untreated garaetteok following storage. Differential scanning calorimetry (DSC) showed that FWRW retarded the retrogradation of garaetteok during storage. The addition of FWRW using Lactobacillus reuteri with an AMG group was particularly effective for inhibiting microbial activity in garaetteok during storage. These results suggest that FWRW using AMG-added L. reuteri can be used as a novel food additive for improving the quality of traditional Korean starch foods and could also reduce the volume of waste WRW.

4.
Bioanalysis ; 14(16): 1111-1122, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-36165918

RESUMEN

Background: Microdialysis sampling after drug microdosing may provide tissue pharmacokinetic data early in clinical drug development. However, low administered doses and small sample volumes pose an analytical challenge, particularly for highly protein-bound drugs. Materials & methods: Carbon-14 [14C]diclofenac was used as a model drug to assess the technical and analytical feasibility of in vivo microdialysis after microdose administration in an in vitro setup. Results: [14C]diclofenac dialysate concentrations were accurately quantified with accelerator MS. [14C]diclofenac dialysate recoveries were similar in the presence and absence of therapeutic diclofenac concentrations but were considerably decreased when albumin was added to the immersion solution, suggesting high protein binding. Conclusion: These results demonstrate the feasibility of combining microdosing and microdialysis to assess tissue pharmacokinetics.


Asunto(s)
Albúminas , Diclofenaco , Radioisótopos de Carbono , Soluciones para Diálisis , Espectrometría de Masas/métodos , Microdiálisis , Preparaciones Farmacéuticas
5.
J Clin Med ; 11(3)2022 Jan 20.
Artículo en Inglés | MEDLINE | ID: mdl-35159967

RESUMEN

BACKGROUND: The purpose of this study was to evaluate the effectiveness of minimally invasive posterior mono-axial pedicle screws fixation in the treatment of thoracolumbar burst fractures. METHODS: In the present study, we analyzed 98 patients retrospectively who had thoracolumbar burst fractures without a neurological deficit. Patients were divided into two groups: mono-axial pedicle screw fixation group (n = 52) and poly-axial pedicle screw fixation group (n = 46). We collected clinical data (visual analog scale (VAS) score for back pain) and included radiographic measurements. RESULTS: Sagittal index was significantly improved at postop and last follow-up in the mono group and the poly group. The mono group was better for reducing and maintaining anterior vertebral height. For the mono group, the mean postoperative regional kyphosis correction rate was 62.31%, and correction loss was 14.18% in late follow-up. For the poly group, the mean postoperative regional kyphosis correction rate was 52.17%, and correction loss was 33.42% in late follow-up. The mono-axial pedicle screw group had a good correction rate and reduced the risks of correction loss. The mean VAS scores for back pain improved by 2.4/2.5 and 3.8/4.2 for the mono and poly groups, respectively. There was no significant difference between groups. CONCLUSIONS: The mono-axial pedicle screw fixation was better for reducing and maintaining anterior vertebral height and regional kyphosis. Therefore, the mono-axial pedicle screw is a better optional instrumentation to treat thoracolumbar vertebral fractures.

6.
Foods ; 10(10)2021 Sep 25.
Artículo en Inglés | MEDLINE | ID: mdl-34681318

RESUMEN

The use of unmodified starch in frozen foods can cause extremely undesirable textural changes after the freeze-thaw process. In this study, using cyclodextrin glucanotransferase (CGTase) and branching enzymes, an amylopectin cluster with high freeze-thaw stability was produced, and was named CBAC. It was found to have a water solubility seven times higher, and a molecular weight 77 times lower, than corn starch. According to the results of a differential scanning calorimetry (DSC) analysis, dough containing 5% CBAC lost 19% less water than a control dough after three freeze-thaw cycles. During storage for 7 days at 4 °C, bread produced using CBAC-treated dough exhibited a 14% smaller retrogradation peak and 37% less hardness than a control dough, suggesting that CBAC could be a potential candidate for clean label starch, providing high-level food stability under repeated freeze-thaw conditions.

7.
Phytomedicine ; 92: 153763, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34601222

RESUMEN

BACKGROUND: Misfolded proteins are formed in the endoplasmic reticulum (ER) due to diverse stimuli including oxidant production, calcium disturbance, and inflammatory factors. Accumulation of these non-native proteins in the ER evokes cellular stress involving the activation of unfolded protein response (UPR) and the execution of ER-associated degradation (ERAD). Naturally-occurring plant compounds are known to interfere with UPR due to their antioxidant and anti-inflammatory activities, leading to inhibition of ER stress. However, there are few studies dealing with the protective effects of natural compounds on the functionality of ERAD. PURPOSE: The current study examined whether asaronic acid enhanced ubiquitin-proteasomal degradation in J774A.1 murine macrophages exposed to 7ß-hydroxycholesterol, a risk factor for atherosclerosis. Asaronic acid (2,4,5-trimethoxybenzoic acid), identified as one of purple perilla constituents, has anti-diabetic and anti-inflammatory effects. Little is known regarding the effects of asaronic acid on the ERAD process and the ubiquitin-proteasomal degradation. METHODS AND RESULTS: Murine macrophages were incubated with 28 µM 7ß-hydroxycholesterol in absence and presence of 1-20 µΜ asaronic acid for up to 24 h. Nontoxic asaronic acid in macrophage diminished the activation of the ER stress sensors of ATF6, IRE1 and PERK stimulated by 7ß-hydroxycholesterol. This methoxybenzoic acid down-regulated the oxysterol-induced expression of EDEM1, OS9, Sel1L-Hrd1 and p97/VCP1, all required for the recognition, recruitment and dislocation of misfolded proteins. On the other hand, asaronic acid enhanced the ubiquitin-proteasomal degradation of non-native proteins dislocated to the cytosol by 7ß-hydroxycholesterol, which entailed the induction of the chaperones of Hsp70 and CHIP and the increased colocalization of ubiquitin and proteasomes. Taken together, asaronic acid attenuated the induction of the UPR-associated sensors and the dislocation-linked transmembrane components in the ER. Conversely, this compound enhanced the proteasomal degradation of dislocated non-native proteins in concert with the chaperones of Hsp70 and CHIP through ubiquitination. CONCLUSION: These observations demonstrate that asaronic acid may be a potent atheroprotective agent as a natural chaperone targeting ER stress-associated macrophage injury.


Asunto(s)
Hidroxicolesteroles , Ubiquitina , Animales , Estrés del Retículo Endoplásmico , Degradación Asociada con el Retículo Endoplásmico , Macrófagos , Ratones
8.
Artículo en Inglés | MEDLINE | ID: mdl-34527066

RESUMEN

Pueraria lobata (Willd.) Ohwi, known as kudzu, is one of the most popular traditional medicines in Asian countries. It has been widely used as a natural alternative to hormone replacement therapy for treating postmenopausal symptoms. This study aimed to investigate the estrogenic effect of P. lobata extract (PE) against postmenopausal osteoporosis in ovariectomized (OVX) rats. OVX rats were treated with PE (25-1600 mg/kg) for 8 weeks. Biochemical parameters, estradiol, and bone turnover markers (e.g., osteocalcin, C-terminal telopeptide fragment of type I collagen, deoxypyridinoline, and pyridinoline) were measured in plasma samples. In addition, estrogen receptor-alpha (ER-α) protein expression and morphology of uterine were evaluated. Long-term treatment with PE did not cause liver damage in OVX rats. PE supplementation reduced body weight gain in obese rats with high lipid accumulation induced by ovariectomy. Furthermore, PE exhibited a protective effect against insulin resistance, hyperlipidemia, and hepatic lipid peroxidation. PE treatment increased uterine weight and thickness of the uterine layers in cases of uterus atrophy due to removal of ovaries. The levels of bone turnover markers, which were significantly increased in OVX rats, were decreased by PE treatment. Western blotting analysis showed that ER-α protein expression was upregulated in PE-treated rats compared with OVX rats. These results suggest that PE could be a promising alternative functional food for improving menopausal symptoms.

9.
Int J Mol Sci ; 22(7)2021 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-33916310

RESUMEN

Epidemiological evidence shows that smoking causes a thrombophilic milieu that may play a role in the pathophysiology of chronic obstructive pulmonary disease (COPD) as well as pulmonary thromboembolism. The increased nicotine level induces a prothrombotic status and abnormal blood coagulation in smokers. Since several anticoagulants increase bleeding risk, alternative therapies need to be identified to protect against thrombosis without affecting hemostasis. Astragalin is a flavonoid present in persimmon leaves and green tea seeds and exhibits diverse activities of antioxidant and anti-inflammation. The current study investigated that astragalin attenuated smoking-induced pulmonary thrombosis and alveolar inflammation. In addition, it was explored that molecular links between thrombosis and inflammation entailed protease-activated receptor (PAR) activation and oxidative stress-responsive mitogen-activated protein kinase (MAPK)-signaling. BALB/c mice were orally administrated with 10-20 mg/kg astragalin and exposed to cigarette smoke for 8 weeks. For the in vitro study, 10 U/mL thrombin was added to alveolar epithelial A549 cells in the presence of 1-20 µM astragalin. The cigarette smoking-induced the expression of PAR-1 and PAR-2 in lung tissues, which was attenuated by the administration of ≥10 mg/kg astragalin. The oral supplementation of ≥10 mg/kg astragalin to cigarette smoke-challenged mice attenuated the protein induction of urokinase plasminogen activator, plasminogen activator inhibitor-1and tissue factor, and instead enhanced the induction of tissue plasminogen activator in lung tissues. The astragalin treatment alleviated cigarette smoke-induced lung emphysema and pulmonary thrombosis. Astragalin caused lymphocytosis and neutrophilia in bronchoalveolar lavage fluid due to cigarette smoke but curtailed infiltration of neutrophils and macrophages in airways. Furthermore, this compound retarded thrombin-induced activation of PAR proteins and expression of inflammatory mediators in alveolar cells. Treating astragalin interrupted PAR proteins-activated reactive oxygen species production and MAPK signaling leading to alveolar inflammation. Accordingly, astragalin may interrupt the smoking-induced oxidative stress-MAPK signaling-inflammation axis via disconnection between alveolar PAR activation and pulmonary thromboembolism.


Asunto(s)
Quempferoles/uso terapéutico , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Embolia Pulmonar/prevención & control , Enfisema Pulmonar/prevención & control , Receptores Proteinasa-Activados/antagonistas & inhibidores , Animales , Fumar Cigarrillos/efectos adversos , Evaluación Preclínica de Medicamentos , Quempferoles/farmacología , Masculino , Ratones Endogámicos BALB C , Estrés Oxidativo , Embolia Pulmonar/etiología
10.
Food Chem ; 345: 128777, 2021 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-33321347

RESUMEN

Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) produces cycloamyloses (CAs), which are large cyclic glucans, and subsequently transforms them to α-, ß-, and γ-cyclodextrins. We developed a novel encapsulation process based on the cyclization activity of CGTase and applied it to the formation of CA inclusion complexes with resveratrol (RVT), which has limited bioavailability due to its low water solubility. The encapsulated RVT (CA-RVT) was purified using preparative high-performance liquid chromatography. The water solubility of CA-RVT was 6,000-fold higher than that of RVT. CA-RVT in water demonstrated 98% stability for 1 week at 4 °C. According to radical scavenging activity and anti-inflammatory assays, CA-RVT in aqueous solution exhibited similar activities as an equal amount of RVT in dimethyl sulfoxide, suggesting the limited solubility of RVT can be overcome through CA encapsulation by CGTase, thus enhancing its nutraceutical value as a functional ingredient in the food industry.


Asunto(s)
Ciclodextrinas/química , Glucosiltransferasas/metabolismo , Resveratrol/química , Cápsulas , Ciclización , Ciclodextrinas/metabolismo , Solubilidad , Agua/química
11.
Knee Surg Sports Traumatol Arthrosc ; 29(12): 4022-4031, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32975624

RESUMEN

PURPOSE: Suprascapular nerve block (SSNB) is the most commonly used block for the relief of postoperative pain from arthroscopic rotator cuff repair and can be used in combination with axillary nerve block (ANB). Dexmedetomidine (DEX) is a type of alpha agonist that can elongate the duration of regional block. The aim of this study was to compare the effects of the use of dexmedetomidine combined with SSNB and ANB with those of the use of SSNB and ANB alone on postoperative pain, satisfaction, and pain-related cytokines within the first 48 h after arthroscopic rotator cuff repair. METHODS: Forty patients with rotator cuff tears who had undergone arthroscopic rotator cuff repair were enrolled in this single-center, double-blinded randomized controlled trial study. Twenty patients were randomly allocated to group 1 and received ultrasound-guided SSNB and ANB using a mixture of 0.5 ml (50 µg) of DEX and 9.5 ml of 0.75% ropivacaine preemptively. The other 20 patients were allocated to group 2 and underwent ultrasound-guided SSNB and ANB alone using a mixture of 0.5 ml of normal saline and 9.5 ml of ropivacaine. The visual analog scale (VAS) for pain and patient satisfaction (SAT) scores were postoperatively checked within 48 h. The plasma interleukin (IL)-6, IL-8, IL-1ß, cortisol, and serotonin levels were also postoperatively measured within 48 h. RESULTS: Group 1 showed a significantly lower mean VAS (visual analog scale of pain) score 1, 3, 6, 12, 18 and 24 h after operation, and a significantly higher mean SAT (patient satisfaction) score 1, 3, 6, 12, 18, 24 and 36 h after the operation than group 2. Group 1 showed a significantly lower mean plasma IL-8 level 1 and 48 h after the operation, and a significantly lower mean IL-1ß level 48 h after the operation than group 2. Group 1 showed a significantly lower mean plasma serotonin level 12 h after the operation than group 2. The mean timing of rebound pain in group 1 was significantly later than that in group 2 (36 h > 23 h, p = 0.007). Six patients each in groups 1 and 2 showed rebound pain. The others did not show rebound pain. CONCLUSIONS: Ultrasound-guided SSNA and ANB with DEX during arthroscopic rotator cuff repair resulted in a significantly lower mean VAS score and a significantly higher mean SAT score within 48 h after the operation than SSNB and ANB alone. Additionally, SSNB and ANB with DEX tended to result in a later mean timing of rebound pain accompanied by significant changes in IL-8, IL-1ß, and serotonin levels within 48 h after the operation. The present study could provide the basis for selecting objective parameters of postoperative pain in deciding the optimal use of medication for relieving pain. LEVEL OF EVIDENCE: Level I. TRIAL REGISTRATION: 2015-20, ClinicalTrials.gov Identifier: NCT04398589. IRB NUMBER: 2015-20, Hallym University Chuncheon Sacred Heart Hospital.


Asunto(s)
Dexmedetomidina , Bloqueo Nervioso , Lesiones del Manguito de los Rotadores , Anestésicos Locales , Artroscopía , Humanos , Dolor Postoperatorio/tratamiento farmacológico , Manguito de los Rotadores/cirugía , Lesiones del Manguito de los Rotadores/cirugía , Resultado del Tratamiento
12.
Arch Orthop Trauma Surg ; 141(3): 477-488, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33184700

RESUMEN

PURPOSE: Few large-scale studies using adjusted data from national registries have explored the risk factors of subsequent revision in patients with unicompartmental knee arthroplasty (UKA) compared to those with total knee arthroplasty (TKA). We investigated the incidence rate and risk factors of subsequent revision in patients with UKA and TKA. METHODS: We enrolled all patients who had undergone TKA or UKA as the primary surgical procedure without histories of having undergone either procedure during the preceding 2 years. Matched Cox regression models were used to compare the risks of revision between groups after propensity score matching. Revision was defined as conversion to revision TKA after primary TKA and conversion to TKA after UKA. RESULTS: The study enrolled 418,806 TKA patients and 446,009 UKA patients. The risk of revision during the entire study period was higher for patients with UKA than for patients with TKA (adjusted hazard ratio [HR] 1.22, 95% confidence interval [95% CI]: 1.10-1.36). The Kaplan-Meier 8-year survival was 98.7% in the TKA group and 96.7% in the UKA group. Patients with UKA were at an increased risk of revision in cases of advanced age (70-79 years, HR 1.40, 95% CI: 1.15-1.71), female sex (HR 1.32, 95% CI: 1.16-1.49), the presence of chronic obstructive pulmonary disease (COPD) (HR 1.27, 95% CI: 1.05-1.54), the presence of peptic ulcer disease (PUD) (HR 1.34, 95% CI: 1.11-1.61) compared to patients with TKA. In patients with hemiplegia, however, UKA were associated with a lower risk of subsequent revision (HR 0.25, 95% CI: 0.07-0.94). CONCLUSION: The risk of a complete exchange or failure was higher for patients with UKA than for patients with TKA. The most significant independent risk factors for subsequent a complete exchange or failure in patients with UKA were advanced age (70-79 years), female sex, and the presence of comorbidities such as COPD and PUD.


Asunto(s)
Artroplastia de Reemplazo de Rodilla , Anciano , Artroplastia de Reemplazo de Rodilla/efectos adversos , Artroplastia de Reemplazo de Rodilla/métodos , Artroplastia de Reemplazo de Rodilla/mortalidad , Artroplastia de Reemplazo de Rodilla/estadística & datos numéricos , Estudios de Cohortes , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Falla de Prótesis , Reoperación/estadística & datos numéricos , República de Corea , Factores de Riesgo
13.
Food Sci Biotechnol ; 29(5): 667-674, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32419965

RESUMEN

To determine the physiochemical properties of the 4-α-glucanotransferase from Bifidobacterium sp., the bllj_0114 gene encoding 4-α-glucanotransferase was cloned from Bifidobacterium longum subsp. longum JCM 1217 and expressed in Escherichia coli. The amino acid sequence alignment indicated that the recombinant protein, named BL-αGTase, belongs to the glycoside hydrolase (GH) family 77. BL-αGTase was purified using nickel-nitrilotriacetic acid affinity chromatography and characterized using various substrates. The enzyme catalyzed the disproportionation activity, which transfers a glucosyl unit from oligosaccharides to acceptor molecules, and had the highest activity at 40 °C and pH 6.0. In the presence of 5 mM metal ions, in particular Cu2+, Zn2+, and Fe2+, BL-αGTase activity was reduced. To determine whether BL-αGTase can be used to generate thermoreversible gels, potato starch was treated with BL-αGTase for various reaction times. The BL-αGTase-treated starches showed sol-gel reversibility and melted at 59.6-75.7 °C.

14.
Int J Biol Macromol ; 152: 727-734, 2020 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-32092418

RESUMEN

Bacteroides thetaiotaomicron (B. thetaiotaomicron), which resides in the human intestinal tract, has a number of carbohydrate enzymes, including glycoside hydrolase (GH) family 97. Only a few GH 97 enzymes have been characterized to date. In this study, a novel α-galactosidase (Bt_3294) was cloned from B. thetaiotaomicron, expressed in Escherichia coli, and purified using affinity chromatography. This novel enzyme showed optimal activity at 60 °C and pH 7.0. Enzyme activity was reduced by 94.4% and 95.7% in the presence of 5 mM Ca2+ and Fe2+, respectively. It is interesting that Bt_3294 specifically hydrolyzed shorter α-galactosyl oligosaccharides, such as melibiose and raffinose. The D-values of Bt_3294 at 40 °C and 50 °C were about 107 and 6 min, respectively. After immobilization of Bt_3294, the D-values at 40 °C and 50 °C were about 37.6 and 29.7 times higher than those of the free enzyme, respectively. As a practical application, the immobilized Bt_3294 was used to hydrolyze raffinose family oligosaccharides (RFOs) in soy milk, decreasing the RFOs by 98.9%.


Asunto(s)
Bacteroides thetaiotaomicron/metabolismo , Enzimas Inmovilizadas/metabolismo , Glicósido Hidrolasas/metabolismo , alfa-Galactosidasa/metabolismo , Concentración de Iones de Hidrógeno , Melibiosa/metabolismo , Oligosacáridos/metabolismo , Rafinosa/metabolismo , Leche de Soja/metabolismo
15.
J Vis Exp ; (151)2019 09 18.
Artículo en Inglés | MEDLINE | ID: mdl-31609345

RESUMEN

Studies in the field of angiogenesis have been aggressively growing in the last few decades with the recognition that angiogenesis is a hallmark of more than 50 different pathological conditions, such as rheumatoid arthritis, oculopathy, cardiovascular diseases, and tumor metastasis. During angiogenesis drug development, it is crucial to use in vitro assay systems with appropriate cell types and proper conditions to reflect the physiologic angiogenesis process. To overcome limitations of current in vitro angiogenesis assay systems using mainly endothelial cells, we developed a 3-dimensional (3D) co-culture spheroid sprouting assay system. Co-culture spheroids were produced by two human vascular cell precursors, endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs) with a ratio of 5 to 1. ECFCs+MSCs spheroids were embedded into type I collagen matrix to mimic the in vivo extracellular environment. A real-time cell recorder was utilized to continuously observe the progression of angiogenic sprouting from spheroids for 24 h. Live cell fluorescent labeling technique was also applied to tract the localization of each cell type during sprout formation. Angiogenic potential was quantified by counting the number of sprouts and measuring the cumulative length of sprouts generated from the individual spheroids. Five randomly-selected spheroids were analyzed per experimental group. Comparison experiments demonstrated that ECFCs+MSCs spheroids showed greater sprout number and cumulative sprout length compared with ECFCs-only spheroids. Bevacizumab, an FDA-approved angiogenesis inhibitor, was tested with the newly-developed co-culture spheroid assay system to verify its potential to screen anti-angiogenic drugs. The IC50 value for ECFCs+MSCs spheroids compared to the ECFCs-only spheroids was closer to the effective plasma concentration of bevacizumab obtained from the xenograft tumor mouse model. The present study suggests that the 3D ECFCs+MSCs spheroid angiogenesis assay system is relevant to physiologic angiogenesis, and can predict an effective plasma concentration in advance of animal experiments.


Asunto(s)
Células Endoteliales , Células Madre Mesenquimatosas , Neovascularización Fisiológica , Inhibidores de la Angiogénesis/farmacología , Animales , Bevacizumab/farmacología , Técnicas de Cocultivo , Células Endoteliales/citología , Humanos , Ratones , Neovascularización Fisiológica/efectos de los fármacos , Esferoides Celulares/efectos de los fármacos
16.
J Clin Invest ; 129(12): 5381-5399, 2019 12 02.
Artículo en Inglés | MEDLINE | ID: mdl-31487270

RESUMEN

Oral squamous cell carcinoma (OSCC) frequently invades the maxillary or mandibular bone, and this bone invasion is closely associated with poor prognosis and survival. Here, we show that CCL28 functions as a negative regulator of OSCC bone invasion. CCL28 inhibited invasion and epithelial-mesenchymal transition (EMT), and its inhibition of EMT was characterized by induced E-cadherin expression and reduced nuclear localization of ß-catenin in OSCC cells with detectable RUNX3 expression levels. CCL28 signaling via CCR10 increased retinoic acid receptor-ß (RARß) expression by reducing the interaction between RARα and HDAC1. In addition, CCL28 reduced RANKL production in OSCC and osteoblastic cells and blocked RANKL-induced osteoclastogenesis in osteoclast precursors. Intraperitoneally administered CCL28 inhibited tumor growth and osteolysis in mouse calvaria and tibia inoculated with OSCC cells. RARß expression was also increased in tumor tissues. In patients with OSCC, low CCL28, CCR10, and RARß expression levels were highly correlated with bone invasion. Patients with OSCC who had higher expression of CCL28, CCR10, or RARß had significantly better overall survival. These findings suggest that CCL28, CCR10, and RARß are useful markers for the prediction and treatment of OSCC bone invasion. Furthermore, CCL28 upregulation in OSCC cells or CCL28 treatment can be a therapeutic strategy for OSCC bone invasion.


Asunto(s)
Huesos/patología , Quimiocinas CC/farmacología , Neoplasias de la Boca/patología , Receptores de Ácido Retinoico/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/patología , Animales , Línea Celular Tumoral , Transición Epitelial-Mesenquimal , Histona Desacetilasa 1/fisiología , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Invasividad Neoplásica , Osteoclastos/citología , Ligando RANK/fisiología , Receptores CCR10/fisiología , Receptor alfa de Ácido Retinoico/fisiología
17.
Biosens Bioelectron ; 141: 111283, 2019 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-31295707

RESUMEN

In this paper, a 3-coil inductive WPT system having circular shaped external coil and miniaturized spherical shaped implantable coil is designed and optimized at 13.56 MHz to overcome reduced PTE under misalignment conditions. The external coil is placed at a distance of 10 mm from the implantable coil. The proposed design is referred to as C-S WPT system. To evaluate the performance of the C-S WPT system, a 3-coil inductive WPT system having circular shaped external coil and circular shaped implantable coil is designed and is referred to as C-C WPT system. The impacts of perfect alignment, lateral and/or angular misalignments on the overall PTE of the systems are analyzed and compared. At perfect alignment, the PTE of C-C and C-S WPT systems are 59.8% and 52.6%, respectively. Though the PTE of C-C WPT system is slightly higher than C-S WPT system, there is a sharp decrease in PTE of C-C system for angles greater than 40°. At 90° of angular misalignment, the PTE's of C-C and C-S WPT systems are 0.4% and 5.6% respectively. When the coils are laterally displaced by a distance of 10 mm, the C-C and C-S WPT systems produce PTE of 33.4% and 27.1% respectively. The C-C and C-S WPT systems attain PTE of 1.6% and 5.5% respectively at 10 mm displacement and 50° rotation. The simulated systems are fabricated to analyze the result in real-environment. The measurement results show that the C-S WPT system provides better PTE when compared to C-C WPT system under angular and a combination of angular and lateral misalignments.


Asunto(s)
Prótesis e Implantes , Técnicas Biosensibles , Suministros de Energía Eléctrica , Humanos , Diseño de Prótesis , Tecnología Inalámbrica
18.
Arch Oral Biol ; 96: 46-51, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30172945

RESUMEN

OBJECTIVE: Remodeling of alveolar bone is controlled by osteoclast-mediated bone resorption and osteoblast-induced bone formation. LPS of Porphyromonas gingivalis, a major causative agent of periodontitis, produces proinflammatory cytokines in host immune cells, which thereby triggers osteoclastogenesis and leads to alveolar bone resorption. We investigated the anti-periodontitis potential of Platycarya strobilacea leaf extract (PLE), which is used as a traditional medicine in Asian countries. DESIGN: TNF-α levels in cell culture media were measured using a commercially available enzyme-linked immunosorbent assay kit. Osteoclast differentiation was observed by tartrate-resistant acid phosphatase staining, and the expression levels of osteoclastogenic genes were measured by quantitative real-time PCR. Bone-resorbing activity was confirmed by the resorption pit formation, gelatin zymographic, and the cathepsin K activity assays. Osteogenic differentiation was confirmed with an ALP activity assay and alizarin red S staining. RESULTS: PLE treatment inhibited the production of TNF-α in P. gingivalis LPS-stimulated RAW264.7 macrophages. In bone marrow-derived macrophages serving as osteoclast precursors, PLE treatment blocked RANKL-induced osteoclastogenesis and gene expression levels of the osteoclastogenic transcription factor NFATc1, DC-STAMP for osteoclast fusion, and cathepsin K for osteoclast activity. In addition, PLE treatment reduced the formation of resorption pits and the secretion of MMP 9 and cathepsin K from the differentiated osteoclasts. Furthermore, PLE treatment induced osteogenesis by increasing ALP activity and calcium content in preosteoblastic cells. CONCLUSION: PLE inhibits P. gingivalis LPS-induced TNF-α production and bone resorption and induces bone formation. PLE may be a beneficial agent to promote oral health by inhibiting periodontitis-induced alveolar bone loss.


Asunto(s)
Pérdida de Hueso Alveolar/prevención & control , Juglandaceae/clasificación , Extractos Vegetales/farmacología , Hojas de la Planta , Porphyromonas gingivalis/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Pérdida de Hueso Alveolar/microbiología , Animales , Diferenciación Celular/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Lipopolisacáridos/farmacología , Masculino , Ratones , Osteoclastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la Polimerasa
19.
Lab Anim Res ; 34(4): 248-256, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30671112

RESUMEN

O-2-18F-fluoroethyl-l-tyrosine ([18F]FET) has been widely used for glioblastomas (GBM) in clinical practice, although evaluation of its applicability in non-clinical research is still lacking. The objective of this study was to examine the value of [18F]FET for treatment evaluation and prognosis prediction of anti-angiogenic drug in an orthotopic mouse model of GBM. Human U87MG cells were implanted into nude mice and then bevacizumab, a representative anti-angiogenic drug, was administered. We monitored the effect of anti-angiogenic agents using multiple imaging modalities, including bioluminescence imaging (BLI), magnetic resonance imaging (MRI), and positron emission tomography-computed tomography (PET/CT). Among these imaging methods analyzed, only [18F]FET uptake showed a statistically significant decrease in the treatment group compared to the control group (P=0.02 and P=0.03 at 5 and 20 mg/kg, respectively). This indicates that [18F]FET PET is a sensitive method to monitor the response of GBM bearing mice to anti-angiogenic drug. Moreover, [18F]FET uptake was confirmed to be a significant parameter for predicting the prognosis of anti-angiogenic drug (P=0.041 and P=0.007, on Days 7 and 12, respectively, on Pearson's correlation; P=0.048 and P=0.030, on Days 7 and 12, respectively, on Cox regression analysis). However, results of BLI or MRI were not significantly associated with survival time. In conclusion, this study suggests that [18F]FET PET imaging is a pertinent imaging modality for sensitive monitoring and accurate prediction of treatment response to anti-angiogenic agents in an orthotopic model of GBM.

20.
J Phys Ther Sci ; 29(4): 618-621, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28533596

RESUMEN

[Purpose] To investigate the relationship between the lateral tibial rotation angle during knee joint flexion and the medial and lateral hamstring muscle thickness ratio during knee joint extension while resting, doing nothing, in upright standing position. The lateral tibial torsion is an important factor of orthopedic knee joint diseases as well as other weight bearing joint diseases such as osteoarthritis, meniscus syndrome, anterior cruciate ligament rupture, etc. [Subjects and Methods] Thirty healthy young adults participated in this study. The thickness of the medial and lateral hamstrings was measured using ultrasonographic imaging technique during knee extension in a resting position. The angle of tibial rotation was measured with 2D motion analysis during knee flexion in a half kneeling position. Pearson's correlation coefficient was used to test the relationship. [Results] There is no significant relationship between the angle of lateral tibial rotation and the ratio of hamstring muscle thickness. [Conclusion] These results demonstrate that lateral tibial rotation is not affected by hamstrings during rest in a standing position.

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