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1.
Connect Tissue Res ; 64(5): 505-515, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37247252

RESUMO

AIM: Inflammation is a complex host response to harmful infection or injury, and it seems to play a crucial role in tissue regeneration both positively and negatively. We have previously demonstrated that the activation of the complement C5a pathway affects dentin-pulp regeneration. However, limited information is available to understand the role of the complement C5a system related to inflammation-mediated dentinogenesis. The aim of this study was to determine the role of complement C5a receptor (C5aR) in regulating lipopolysaccharide (LPS)-induced odontogenic differentiation of dental pulp stem cells (DPSCs). MATERIAL AND METHODS: Human DPSCs were subjected to LPS-stimulated odontogenic differentiation in dentinogenic media treated with the C5aR agonist and antagonist. A putative downstream pathway of the C5aR was examined using a p38 mitogen-activated protein kinase (p38) inhibitor (SB203580). RESULTS: Our data demonstrated that inflammation induced by the LPS treatment potentiated DPSC odontogenic differentiation and that this is C5aR dependent. C5aR signaling controlled the LPS-stimulated dentinogenesis by regulating the expression of odontogenic lineage markers like dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP-1). Moreover, the LPS treatment increased the total p38, and the active form of p38 expression, and treatment with SB203580 abolished the LPS-induced DSPP and DMP-1 increase. CONCLUSIONS: These data suggest a significant role of C5aR and its putative downstream molecule p38 in the LPS-induced odontogenic DPSCs differentiation. This study highlights the regulatory pathway of complement C5aR/p38 and a possible therapeutic approach for improving the efficiency of dentin regeneration during inflammation.


Assuntos
Polpa Dentária , Lipopolissacarídeos , Humanos , Diferenciação Celular/fisiologia , Proliferação de Células , Células Cultivadas , Complemento C5a/metabolismo , Polpa Dentária/metabolismo , Inflamação/induzido quimicamente , Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Regeneração , Células-Tronco/metabolismo , Proteína Quinase 14 Ativada por Mitógeno/metabolismo
2.
Int J Mol Sci ; 24(22)2023 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-38003452

RESUMO

The recent expiration of patents for the antibiotic tulathromycin has led to a significant increase in the number of generic tulathromycin products (GTPs) available. This study aims to evaluate the bioequivalence of four GTPs, which experienced a rapid increase in market share. The bioequivalence was evaluated by performing pharmacokinetic assessments. The four selected GTPs (Tulaject, Tulagen, Toulashot, and T-raxxin) were compared with the reference product, Draxxin. A dose of 2.5 mg/kg.bw/day was administered via subcutaneous injection, and blood samples were collected 460 times from 20 Holstein cattle. Plasma concentrations of tulathromycin were measured over time using LC-MS/MS analysis. Bioequivalence was evaluated using a statistical program for pharmacokinetic parameters, including the area under the concentration time curve (AUC) and the maximum plasma concentration (Cmax). The bioequivalence was considered proven if the difference between the test and reference products was within 20% for both AUC and Cmax. The results showed that the confidence interval (CI, 90%) for both AUC and Cmax values was within the 80~120% range, demonstrating the bioequivalence of the four GTPs compared to Draxxin. This study provides evidence for the bioequivalence of the selected GTPs, contributing to their validation for use as effective antibiotics.


Assuntos
Compostos Heterocíclicos , Espectrometria de Massas em Tandem , Bovinos , Animais , Cromatografia Líquida , Dissacarídeos , Medicamentos Genéricos/farmacocinética , Área Sob a Curva , Estudos Cross-Over
3.
BMC Microbiol ; 20(1): 118, 2020 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-32410630

RESUMO

BACKGROUND: The antibiotics generally used in farm animals are rapidly losing their effectiveness all over the world as bacteria develop antibiotic resistance. Like some other pathogenic bacteria multidrug-resistant strains of Salmonella enterica serovar Typhimurium (S. Typhimurium) are also frequently found in animals and humans which poses a major public health concern. New strategies are needed to block the development of resistance and to prolong the life of traditional antibiotics. Thus, this study aimed to increase the efficacy of existing antibiotics against S. Typhimurium by combining them with opportunistic phenolic compounds gallic acid (GA), epicatechin, epicatechin gallate, epigallocatechin and hamamelitannin. Fractional inhibitory concentration indexes (FICI) of phenolic compound-antibiotic combinations against S. Typhimurium were determined. Based on the FICI and clinical importance, 1 combination (GA and ceftiofur) was selected for evaluating its effects on the virulence factors of this bacterium. Viability of Rattus norvegicus (IEC-6) cell in presence of this antibacterial combination was evaluated. RESULTS: Minimum inhibitory concentrations (MICs) of GA, epigallocatechin and hamamelitannin found against different strains of S. Typhimurium were 256, (512-1024), and (512-1024) µg/mL, respectively. Synergistic antibacterial effect was obtained from the combination of erythromycin-epicatechin gallate (FICI: 0.50) against S. Typhimurium. Moreover, additive effects (FICI: 0.502-0.750) were obtained from 16 combinations against this bacterium. The time-kill assay and ultrastructural morphology showed that GA-ceftiofur combination more efficiently inhibited the growth of S. Typhimurium compared to individual antimicrobials. Biofilm viability, and swimming and swarming motilities of S. Typhimurium in presence of GA-ceftiofur combination were more competently inhibited than individual antimicrobials. Viabilities of IEC-6 cells were more significantly enhanced by GA-ceftiofur combinations than these antibacterials alone. CONCLUSIONS: This study suggests that GA-ceftiofur combination can be potential medication to treat S. Typhimurium-associated diarrhea and prevent S. Typhimurium-associated blood-stream infections (e.g.: fever) in farm animals, and ultimately its transmission from animal to human. Further in vivo study to confirm these effects and safety profiles in farm animal should be undertaken for establishing these combinations as medications.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Fenóis/farmacologia , Salmonelose Animal/microbiologia , Salmonella typhimurium/fisiologia , Animais , Animais Domésticos , Biofilmes/crescimento & desenvolvimento , Catequina/análogos & derivados , Catequina/farmacologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cefalosporinas/farmacologia , Combinação de Medicamentos , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Sinergismo Farmacológico , Eritromicina/farmacologia , Ácido Gálico/farmacologia , Testes de Sensibilidade Microbiana , Ratos , Salmonelose Animal/tratamento farmacológico , Salmonella typhimurium/efeitos dos fármacos , Sorogrupo
4.
Vet Res ; 49(1): 101, 2018 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-30286813

RESUMO

Salmonella enterica serovar Typhimurium infects intestinal epithelia and macrophages, which is prevented by inhibiting adhesion and cell invasion. This study aimed to investigate the role of methyl gallate (MG) in adhesion, invasion, and intracellular survival of Salmonella Typhimurium in Caco-2 and RAW 264.7 cells via a gentamicin protection assay, confocal microscopy, and quantitative reverse-transcription polymerase chain reaction. MG (30 µg/mL) inhibited adhesion and invasion of Salmonella Typhimurium by 54.01% and 60.5% in RAW 264.7 cells, respectively. The combination of MG with sub-minimum inhibitory concentration (MIC) of marbofloxacin (MRB) inhibited the adhesion, invasion, and intracellular survival by 70.49%, 67.36%, and 74%, respectively. Confocal microscopy further revealed reductions in bacterial count in Caco-2 cells treated with MG alone or with sub-MIC of MRB. Furthermore, MG alone or in combination with sub-MIC of MRB decreased the motility of Salmonella Typhimurium. Quorum sensing genes including sdiA, srgE, and rck were downregulated by 52.8%, 61.7%, and 22.2%, respectively. Moreover, rac-1 was downregulated by 56.9% and 71.9% for MG alone and combined with sub-MIC of MRB, respectively, in mammalian cells. Furthermore, MG downregulated virulence genes of Salmonella Typhimurium including cheY, ompD, sipB, lexA, and ompF by 59.6%, 60.2%, 20.5%, 31.4%, and 16.2%, respectively. Together, the present results indicate that MG alone or in combination with a sub-MIC of MRB effectively inhibited the adhesion, invasion, and intracellular survival of Salmonella Typhimurium in vitro by downregulating quorum sensing and virulence genes.


Assuntos
Antibacterianos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Fluoroquinolonas/farmacologia , Ácido Gálico/análogos & derivados , Salmonella typhimurium/efeitos dos fármacos , Animais , Proteínas de Bactérias/genética , Células CACO-2 , Regulação para Baixo , Ácido Gálico/farmacologia , Humanos , Camundongos , Células RAW 264.7 , Salmonella typhimurium/genética , Salmonella typhimurium/fisiologia , Virulência/efeitos dos fármacos , Virulência/genética
5.
BMC Complement Altern Med ; 18(1): 222, 2018 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-30041643

RESUMO

BACKGROUND: Medicinal plants represent a source of new drugs for the prevention and treatment of infectious diseases. Dendropanax morbifera Léveille is an economically and medicinally important subtropical tree that has various biological activities. However, its ability to affect immune responses in vivo is unknown. Hence, this study was designed to examine the immunomodulatory activity of fermented D. morbifera extract in BALB/c mice. METHODS: five-week-old female BALB/c mice were arranged in six groups and kept under a standard laboratory condition. Splenocyte counts were determined using the trypan blue dye exclusion method, and splenic lymphocyte proliferation was determined using concanavalin A and lipopolysaccharide (LPS). Flow cytometric analysis was performed to phenotype T-lymphocytes. Next, cytokine and immunoglobulin quantitation was performed using sandwich ELISA. RESULTS: The results showed an increase in spleen cells by 71 and 67% in mice treated with 125 and 250 mg/kg of D. morbifera, respectively. In addition, splenocyte proliferation was increased 58.7% in response to concanavalin A treatment, while LPS treatment induced a 73.3% increase in mice treated with 125 mg/kg. T-cell phenotypic analysis indicated that D. morbifera-treated groups showed higher CD8a+, CD11b and CD3+ T-cell expression. However, the treatment groups showed suppression of IL-1α, Il-1ß and IL-4. In addition, the IgG super-family was downregulated in a dose-dependent manner by 4.5% up to 43.7%. CONCLUSIONS: Taken together, we show that D. morbifera increases the number and proliferation of T- and B-lymphocytes. Moreover, these effects may play a role in boosting non-specific immunity, while suppressing proinflammatory cytokines and immunoglobulins after a single antigen exposure.


Assuntos
Anti-Inflamatórios/farmacologia , Araliaceae/química , Extratos Vegetais/farmacologia , Linfócitos T/efeitos dos fármacos , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/toxicidade , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/análise , Citocinas/metabolismo , Feminino , Fermentação , Imunoglobulina G , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/química , Extratos Vegetais/toxicidade , Folhas de Planta/química , Baço/citologia , Linfócitos T/metabolismo
6.
BMC Vet Res ; 13(1): 136, 2017 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-28532461

RESUMO

BACKGROUND: Veterinary medicines have been widely used for the prevention and treatment of diseases, growth promotion, and to promote feeding efficacy in livestock. As the veterinary medicine industry has steadily grown, it is crucial to set up a baseline for the quality of medicine as well as the insufficiency or excessiveness of the active ingredients in drug products to ensure the compliance, safety and efficacy of these medicines. Thus, the 10 years data of post-marketing quality control study was summarized to determine the rate and extent of non-compliance of these medicines and to establish baseline data for future quality control measures of veterinary medicine. RESULTS: In this study, 1650 drugs for veterinary use were collected per year from each city and province in Korea and analysed for the quantity of active ingredients according to the "national post-market surveillance (NPMS) system" over the past decade. The NPMS assessment was performed using liquid and gas chromatography, titration, UV/Vis spectrophotometry, and bioassays. A total of 358 cases were deemed noncompliant, with the average noncompliance rate for all medicine types being 2.0%. The average noncompliance rates for antibiotics, biologics and other chemical drugs except antibiotics (OCD) were 1.1%, 1.2%, and 3.0%, respectively. The first leading cause for noncompliant products was insufficient quantity of major ingredients (283 cases), and the second leading cause was the existence of excess amount of active ingredients (60 cases). Tylosin, spiramycin, ampicillin, tetracyclines and penicillins were most frequently found to be noncompliant among antibiotics. Among the OCD, the noncompliance was found commonly in vitamin A. CONCLUSION: The overall trend presented gradually decreasing violation rates, suggesting that the quality of veterinary medicines has improved. Consistent application of the NPMS assessment and the establishment of the Korea Veterinary Good Manufacturing Practice (KVGMP) will help to maintain the good quality of medicine.


Assuntos
Vigilância de Produtos Comercializados , Drogas Veterinárias/normas , Garantia da Qualidade dos Cuidados de Saúde , República da Coreia
7.
Fish Shellfish Immunol ; 54: 466-72, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27108377

RESUMO

Wild and farm-raised fish can be simultaneously exposed to different types of pathogens in their habitats. Hence, it is important to study their effects, whether isolated or in combination. Therefore, the aim of this study was to evaluate the effects of Lactobacillus pentosus PL11 on the transcription of specific cytokine genes related to immune response, using Japanese eel macrophages as an in vitro model. Head kidney leukocytes were isolated from Japanese eels and cell viability was determined using an MTT reagent. In addition, the Griess reagent was used to determine the nitric oxide (NO) production while, an enzyme-linked immunosobent assay (ELISA) and a quantitative polymerase chain reaction (qPCR) were utilized to quantify the level of proinflammatory cytokines. The results of the study indicated that infection by Edwardsiella tarda alone causes a higher rate of cell death and an increase in the production of proinflammatory cytokines, such as interleukin-1ß (IL-1ß, 822.67 ± 29.48 pg mL(-1)), interleukin-6 (IL-6, 13.57 ± 0.55 pg mL(-1)), and tumor necrosis factor-α (TNF-α, 2033.67 ± 84.68 pg mL(-1)). However, co-culture with L. pentosus PL11 downregulates the production of NO and the related IL-1ß, IL-6, and TNF-α by 46%, 88.4%, 59%, and 77%, respectively. Quantification of the mRNA expression level revealed it to be consistent with the ELISA analysis. Hence, we infer that L. pentosus PL11 plays a significant role in the immunmodulation of the inflammatory responses that arise in fish owing to infection by pathogenic bacteria such as Edwardsiella tarda.


Assuntos
Anguilla , Edwardsiella tarda/efeitos dos fármacos , Infecções por Enterobacteriaceae/veterinária , Doenças dos Peixes/imunologia , Imunomodulação , Lactobacillus pentosus , Probióticos , Anguilla/imunologia , Anguilla/microbiologia , Ração Animal/análise , Animais , Células Cultivadas , Dieta/veterinária , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/microbiologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Rim Cefálico/microbiologia , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Lactobacillus pentosus/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
8.
Anal Bioanal Chem ; 408(29): 8539-8549, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27744479

RESUMO

Conventional antimicrobial susceptibility tests (ASTs) are very time consuming and insufficiently precise to promptly select a proper antimicrobial treatment. This difficulty disrupts the management of infections and exacerbates the development of antimicrobial resistance. Generally, antimicrobial resistance involves the chemical modification of an antimicrobial compound to an inactive form by an enzyme released by bacteria. This modification causes a structural change and is followed by a characteristic mass shift of the antimicrobials. Using this mechanism, we developed a new liquid chromatography-mass spectrometry method to rapidly determine the degree of resistance of Salmonella enterica subspecies enterica serovar Typhimurium (Salmonella Typhimurium), Escherichia coli, and Staphylococcus aureus to amoxicillin, ampicillin, and penicillin G, respectively. This method was successfully applied to 20 bacterial isolates from Korean slaughterhouses and farms. There were 18-Da mass shifts in resistant strains compared with susceptible strains of Salmonella Typhimurium, E. coli, and S. aureus, and the intensities of the hydrolyzed penicillin mass spectra were much higher in resistant strains than those in susceptible strains, which together indicate the reliability of this method. A comparison of the mass spectrometry-derived results with that from conventional ASTs revealed an identical classification of the tested bacteria according to sensitivity and resistance. Notably, this assay method requires only 2 h for determining the susceptibility status of a strain. This newly developed method is able to determine the extent of antimicrobial resistance qualitatively and quantitatively within a very short time and could be used to replace conventional AST methods. Graphical abstract Rapid determination of ß-lactam antimicrobial resistance in bacteria by LC-MS/MS.


Assuntos
Escherichia coli/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Resistência beta-Lactâmica/efeitos dos fármacos , beta-Lactamas/farmacologia , Animais , Cromatografia Líquida de Alta Pressão/métodos , Escherichia coli/enzimologia , Fezes/microbiologia , Hidrólise , Limite de Detecção , Testes de Sensibilidade Microbiana , Salmonella typhimurium/enzimologia , Staphylococcus aureus/enzimologia , Espectrometria de Massas em Tandem/métodos , beta-Lactamases/metabolismo
9.
Genomics ; 102(1): 47-56, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23628646

RESUMO

In the present study, a computational comparative and subtractive genomic/proteomic analysis aimed at the identification of putative therapeutic target and vaccine candidate proteins from Kyoto Encyclopedia of Genes and Genomes (KEGG) annotated metabolic pathways of Mycoplasma hyopneumoniae was performed for drug design and vaccine production pipelines against M.hyopneumoniae. The employed comparative genomic and metabolic pathway analysis with a predefined computational systemic workflow extracted a total of 41 annotated metabolic pathways from KEGG among which five were unique to M. hyopneumoniae. A total of 234 proteins were identified to be involved in these metabolic pathways. Although 125 non homologous and predicted essential proteins were found from the total that could serve as potential drug targets and vaccine candidates, additional prioritizing parameters characterize 21 proteins as vaccine candidate while druggability of each of the identified proteins evaluated by the DrugBank database prioritized 42 proteins suitable for drug targets.


Assuntos
Biologia Computacional , Desenho de Fármacos , Redes e Vias Metabólicas/genética , Mycoplasma hyopneumoniae/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bases de Dados de Compostos Químicos , Genoma Bacteriano , Humanos , Anotação de Sequência Molecular , Mycoplasma hyopneumoniae/efeitos dos fármacos , Mycoplasma hyopneumoniae/patogenicidade , Vacinas Sintéticas/genética , Vacinas Sintéticas/metabolismo
10.
Pol J Vet Sci ; 17(1): 99-103, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24724476

RESUMO

The association between physiologically dependent pharmacokinetic parameters (CL(B), T1/2beta, Vd(ss)) of marbofloxacin and body weight was studied in eight animal species based on allometric equation Y = aWb, where 'Y' is the pharmacokinetic parameter, 'W' is body weight, 'a' is allometric coefficient (intercept) and 'b' is the exponent that describes relation between pharmacokinetic parameter and body weight. The body clearance of marbofloxacin has shown significant (P < 0.0001) relation with size (Bwt) in various animal species. However, half-life and volume of distribution were not in association with body weight. Although half-life and volume of distribution were not in a good correlation with body weight, statistically significant association between the body clearance and body weight suggests validity of allometric scaling for predicting pharmacokinetic parameters of marbofloxacin in animal species that have not been studied yet. However further study considering large sample size and other parameters influencing pharmacokinetics of marbofloxacin is recommended.


Assuntos
Antibacterianos/farmacocinética , Peso Corporal , Fluoroquinolonas/farmacocinética , Animais , Meia-Vida , Estudos Retrospectivos , Especificidade da Espécie , Distribuição Tecidual
11.
Vet Med Sci ; 9(3): 1292-1296, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-37018122

RESUMO

BACKGROUND: Existing treatments against poultry red mite (PRM; Dermanyssus gallinae) infestation have reduced efficacy or exhibit hazardous effects on chickens. Considering the economic importance of chickens, development of a safe and effective method for exterminating PRMs is necessary. Ivermectin and allicin are effective against some ectoparasites; however, their acaricidal efficacies against PRMs remain unknown. OBJECTIVE: To evaluate individual and combined efficacies of ivermectin and allicin in exterminating PRMs. METHODS: Different concentrations (0.10-1.0 mg/mL) of ivermectin (1 mL) were applied via dropping method in different insect culture dishes (ICDs), prior to transferring PRMs. For the spraying method, PRMs were transferred to ICDs, before spraying ivermectin (1 mg/mL) solution (1 mL). Further, the acaricidal effect of allicin on PRMs was evaluated by applying different concentrations (0.25-1.0 mg/mL) of allicin (1 mL). The combined acaricidal effects of ivermectin and allicin were analysed using four concentration combinations. PRM death rates were determined after 2 h, 24 h, 2 days, 5 days and 7 days of drug application. RESULTS: Ivermectin application (1 mg/mL) exterminated 64% and 100% of PRMs on 1 and 5 days, respectively, and prevented their revival. Further, 0.5 mg/mL ivermectin and 1 mg/mL allicin individually exterminated 98% and 44% of PRMs, respectively, within 7 days of treatment. In combination, 0.5 mg/mL ivermectin and 0.5 mg/mL allicin exterminated 100% of PRMs within 5 d of treatment. The most effective combination was 0.25 mg/mL ivermectin + 1.00 mg/mL allicin. CONCLUSIONS: The efficacy of ivermectin-allicin combination in exterminating PRMs was demonstrated. This novel approach could be optimised for industrial applications.


Assuntos
Acaricidas , Infestações por Ácaros , Ácaros , Doenças das Aves Domésticas , Animais , Aves Domésticas , Ivermectina/uso terapêutico , Ivermectina/farmacologia , Infestações por Ácaros/tratamento farmacológico , Infestações por Ácaros/veterinária , Infestações por Ácaros/parasitologia , Galinhas , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/prevenção & controle , Acaricidas/uso terapêutico , Acaricidas/farmacologia
12.
Cells ; 12(14)2023 07 14.
Artigo em Inglês | MEDLINE | ID: mdl-37508514

RESUMO

The odontoblastic differentiation of dental pulp stem cells (DPSCs) associated with caries injury happens in an inflammatory context. We recently demonstrated that there is a link between inflammation and dental tissue regeneration, identified via enhanced DPSC-mediated dentinogenesis in vitro. Brain-derived neurotrophic factor (BDNF) is a nerve growth factor-related gene family molecule which functions through tropomyosin receptor kinase B (TrkB). While the roles of BDNF in neural tissue repair and other regeneration processes are well identified, its role in dentinogenesis has not been explored. Furthermore, the role of BDNF receptor-TrkB in inflammation-induced dentinogenesis remains unknown. The role of BDNF/TrkB was examined during a 17-day odontogenic differentiation of DPSCs. Human DPSCs were subjected to odontogenic differentiation in dentinogenic media treated with inflammation inducers (LTA or TNFα), BDNF, and a TrkB agonist (LM22A-4) and/or antagonist (CTX-B). Our data show that BDNF and TrkB receptors affect the early and late stages of the odontogenic differentiation of DPSCs. Immunofluorescent data confirmed the expression of BDNF and TrkB in DPSCs. Our ELISA and qPCR data demonstrate that TrkB agonist treatment increased the expression of dentin matrix protein-1 (DMP-1) during early DPSC odontoblastic differentiation. Coherently, the expression levels of runt-related transcription factor 2 (RUNX-2) and osteocalcin (OCN) were increased. TNFα, which is responsible for a diverse range of inflammation signaling, increased the levels of expression of dentin sialophosphoprotein (DSPP) and DMP1. Furthermore, BDNF significantly potentiated its effect. The application of CTX-B reversed this effect, suggesting TrkB`s critical role in TNFα-mediated dentinogenesis. Our studies provide novel findings on the role of BDNF-TrkB in the inflammation-induced odontoblastic differentiation of DPSCs. This finding will address a novel regulatory pathway and a therapeutic approach in dentin tissue engineering using DPSCs.


Assuntos
Receptor trkB , Fator de Necrose Tumoral alfa , Humanos , Fator de Necrose Tumoral alfa/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Receptor trkB/metabolismo , Tropomiosina/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Polpa Dentária , Diferenciação Celular , Inflamação/metabolismo , Células-Tronco
13.
Antibiotics (Basel) ; 12(5)2023 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-37237780

RESUMO

A safe and effective method for eradicating poultry red mite (PRM; Dermanyssus gallinae) is urgently needed, as existing treatments show a low efficacy or hazardous effects on chickens. We evaluated the efficacy of a combined treatment with ivermectin and allicin (IA) against PRMs in chickens and drug residues in non-target samples. The efficiency of PRM eradication by IA was compared with those of natural acaricides in vitro. Ivermectin (0.25 mg/mL) + allicin (1 mg/mL) (IA compound) was sprayed on isolator housing hens with PRMs. The PRM mortality rate, clinical symptoms, and ivermectin residue in hens were analyzed. IA showed the highest PRM-eradication efficacy among all tested compounds in vitro. The insecticidal rates of IA were 98.7%, 98.4%, 99.4%, and 99.9% at 7, 14, 21, and 28 days of treatment, respectively. After inoculating PRMs, hypersensitivity, itching, and a pale-colored comb were observed in control animals, which were absent in treated hens. No clinical symptoms from IA and ivermectin residues were found in hens. IA effectively exterminated PRMs, demonstrating its potential for industrial use to treat PRMs.

14.
Antibiotics (Basel) ; 12(6)2023 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-37370342

RESUMO

Employing affordable and uncomplicated sample preparation techniques to recommend the most efficient antibacterial therapy could help reduce antibiotic-resistant bacteria. This study evaluated the suitability of immunoassays and microbiological assays as alternatives for liquid chromatography/mass spectrometry (LC/MS) in determining plasma tylosin concentrations after intramuscular administration at a dose of 20 mg/kg to both healthy and diseased pigs in clinical veterinary practice. The diseased pigs were confirmed using the target genes Actinobacillus pleuropneumoniae (apxIVA) and Pasteurella multocida (kmt1). The methods showed good linearity, precision, and accuracy. In both healthy and diseased pigs, a significant correlation was observed between LC/MS and the microbiological assay (Pearson correlation coefficient: 0.930, p < 0.001 vs. Pearson correlation coefficient: 0.950, p < 0.001) and between LC/MS and the enzyme-linked immunosorbent assay (ELISA) (Pearson correlation coefficient: 0.933; p < 0.001 vs. Pearson correlation coefficient: 0.976, p < 0.001). A strong correlation was observed between the microbiological assay and the ELISA in both healthy and diseased pigs (Pearson correlation coefficient: 0.911; p < 0.001 vs. Pearson correlation coefficient: 0.908, p < 0.001). A Bland-Altman analysis revealed good agreement between the methods, i.e., 95% of the differences were within the limits of agreement. Therefore, the microbiological assay and the ELISA, which demonstrated sufficient precision and accuracy, can be viable alternatives to LC/MS when it is unavailable.

15.
Antibiotics (Basel) ; 10(2)2021 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-33572763

RESUMO

Poultry meat and eggs are vital sources of protein for human consumption worldwide. The use of several nutritional and medicinal products, including antibiotics, is crucial for efficient and safe poultry production. Accumulation of drug residues in meat and eggs from inappropriate drug use is a major concern to public health. Recently, enrofloxacin was detected (2.4-3.8 ppb) in edible eggs produced in Jeju Island, Korea. Although the farm from which the enrofloxacin-contaminated eggs were collected did not use enrofloxacin-containing products, they reported extensive use of a nutritional product (NPJ). Accordingly, in this study, we investigated whether enrofloxacin contamination had occurred accidentally in various widely used veterinary pharmaceutical products. Enrofloxacin content (4.57-179.08 ppm) in different lots of the NPJ was confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Furthermore, 76 veterinary pharmaceutical products that are widely used in poultry farms in Korea and claim to not contain enrofloxacin were collected and analyzed by LC-MS/MS. Among them, a florfenicol product and a sulfatrimethoprime product were found to contain 3.00 and 0.57 ppm enrofloxacin, respectively. These results suggest that appropriate manufacturing standards are not being followed and that strict monitoring of drug manufacturing is necessary in Korea to avoid drug contamination.

16.
Vet Med Sci ; 7(2): 289-296, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33107216

RESUMO

Veterinary biocides used in animal husbandry have the potential to cause human health concerns. Biocidal products for veterinary use, which contain pesticides approved in Korea, comprise 49 active ingredients within 234 products. Within 17 of these products there are 3 ingredients which are highly hazardous pesticides: coumaphos, dichlorvos and methomyl. In this study, the content of the active ingredients of 160 products sold domestically was investigated. Samples were collected for 119 biocidal products for veterinary use. These were analysed by high-performance liquid chromatography (HPLC) and gas chromatography (GC). Seventeen products were noncompliant (insufficient or excess quantity of active ingredients). The ingredients that were below the stated concentrations were amitraz, chlorpyrifos-methyl, cypermethrin, cyromazine, dichlorvos, fipronil, muscamone and trichlorfon. The ingredients that exceeded the stated concentrations were abamectin, fluvalinate and pyriproxyfen. The noncompliance rate in biocidal products for veterinary use was 9.19%. The results of this study show that three highly hazardous pesticides (coumaphos, dichlorvos and methomyl) and 10 active ingredients (abamectin, amitraz, chlorpyrifos-methyl, cypermethrin, cyromazine, fipronil, fluvalinate, muscamone, pyriproxyfen and trichlorfon) deviated from the stated concentrations. Thus, management plans should be established to ensure compliant veterinary drugs by post-distribution quality control, such as planning for regular inspection.


Assuntos
Praguicidas/análise , Medicina Veterinária/estatística & dados numéricos , Cromatografia Líquida de Alta Pressão/veterinária , República da Coreia
17.
Pathogens ; 9(10)2020 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-33023003

RESUMO

Pathogenic Escherichia coli (E. coli)-associated infections are becoming difficult to treat because of the rapid emergence of antibiotic-resistant strains. Novel approaches are required to prevent the progression of resistance and to extend the lifespan of existing antibiotics. This study was designed to improve the effectiveness of traditional antibiotics against E. coli using a combination of the gallic acid (GA), hamamelitannin, epicatechin gallate, epigallocatechin, and epicatechin. The fractional inhibitory concentration index (FICI) of each of the phenolic compound-antibiotic combinations against E. coli was ascertained. Considering the clinical significance and FICI, two combinations (hamamelitannin-erythromycin and GA-ampicillin) were evaluated for their impact on certain virulence factors of E. coli. Finally, the effects of hamamelitannin and GA on Rattus norvegicus (IEC-6) cell viability were investigated. The FICIs of the antibacterial combinations against E. coli were 0.281-1.008. The GA-ampicillin and hamamelitannin-erythromycin combinations more effectively prohibited the growth, biofilm viability, and swim and swarm motilities of E. coli than individual antibiotics. The concentration of hamamelitannin and GA required to reduce viability by 50% (IC50) in IEC-6 cells was 988.54 µM and 564.55 µM, correspondingly. GA-ampicillin and hamamelitannin-erythromycin may be potent combinations and promising candidates for eradicating pathogenic E. coli in humans and animals.

18.
Infect Drug Resist ; 13: 1601-1606, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32606811

RESUMO

Rapid determination of antimicrobial susceptibility/resistance is an important factor in selecting an appropriate antimicrobial treatment and eradicating infections promptly. Conventional antimicrobial susceptibility tests (ASTs) are very time consuming. Thus, we developed a liquid chromatography-mass spectrometry (LC-MS/MS) method for rapidly determining the resistance of Staphylococcus aureus to penicillin-G in an animal-infection model. This technique will be able to detect those resistant strains whose resistance mechanism specifically controlled by penicillinase. The resistance status of S. aureus against penicillin-G was determined by conventional AST. Cultured S. aureus cells were inoculated to chicken for developing bacteraemia. The solution of penicillin-G was intravenously administered (10 mg/kg b.w.) to chickens just after infection detection. Blood samples were collected at different intervals after drug administration. The concentration of active penicillin-G and its metabolites were determined from the bacteria-free blood supernatant by utilizing the LC-MS/MS method. Evidence of infection in chicken was observed within 5 h of bacterial inoculation. The penicillinase enzyme generated by S. aureus transforms the active penicillin-G to an inactive metabolite by hydrolysis, which is evident by the mass shift from 335.10600 to 353.11579 Da as quantified using liquid chromatography quadrupole time-of-flight mass spectrometry (LC/Q-TOF/MS). The signal intensity of inactive/hydrolysed penicillin-G is several-fold greater than that of the active penicillin-G in the blood sample of chicken infected with resistant strain and treated with penicillin-G. The antimicrobial resistance index (ARI) value of resistant S. aureus strain was more than 1, demonstrating the penicillin-G-resistance pattern of that strain. This method is able to determine the extent of ß-lactam antimicrobial resistance within 1.5 h from the patient's blood and is complementary with those existing AST methods which are usually practicing in the evaluation of ß-lactam antibiotic resistance.

19.
J Vet Sci ; 21(2): e37, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32233140

RESUMO

Poultry red mites (PRMs), Dermanyssus gallinae, are one of the most harmful ectoparasites of laying hens. Because of their public health impact, safe, effective methods to eradicate PRMs are greatly needed. Carbon dioxide (CO2) was shown to eradicate phytophagous mites; however, there is no evidence that PRMs can be eradicated by CO2. Thus, the efficacy of CO2, applied by direct-spraying and dry ice-generated exposure, for eradicating PRMs was investigated. Both treatments eradicated > 85% of PRMs within 24 h and 100% of PRMs by 120 h of post-treatment. Therefore, these novel approaches may be useful for eradicating PRMs in clinical settings.


Assuntos
Dióxido de Carbono/uso terapêutico , Erradicação de Doenças/métodos , Infestações por Ácaros/veterinária , Ácaros , Doenças das Aves Domésticas/prevenção & controle , Animais , Galinhas , Infestações por Ácaros/parasitologia , Infestações por Ácaros/prevenção & controle , Doenças das Aves Domésticas/parasitologia
20.
J Vet Sci ; 20(2): e15, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30944537

RESUMO

It is crucial to optimize the dose of fluoroquinolones to avoid antibiotic resistance and to attain clinical success. We undertook this study to optimize the dose of enrofloxacin against Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) in chicken by assessing its pharmacokinetic/pharmacodynamic (PK/PD) indices. The antibacterial activities of enrofloxacin against S. Enteritidis were evaluated. After administering 10 mg/kg body weight (b.w.) of enrofloxacin to broiler chickens of both sexes by intravenous (IV) and peroral (PO) routes, blood samples were drawn at different intervals and enrofloxacin concentrations in plasma were determined. PK/PD indices were calculated by integrating the PK and PD data. The elimination half-lives (T1/2), time required to reach peak concentration (Tmax), peak concentration (Cmax), and area under curve (AUC) after administering enrofloxacin by PO and IV routes were 25.84 ± 1.40 h, 0.65 ± 0.12 h, 3.82 ± 0.59 µg/mL, and 20.84 ± 5.0 µg·h/mL, and 12.84 ± 1.4 h, 0.22 ± 0.1 h, 6.74 ± 0.03 µg/mL, and 21.13 ± 0.9 µg.h/mL, respectively. The bioavailability of enrofloxacin was 98.6% ± 8.9% after PO administration. The MICs of enrofloxacin were 0.0625-1 µg/mL against S. Enteritidis strains, and the MIC50 was 0.50 µg/mL. The Cmax/MIC50 were 7.64 ± 0.2 and 13.48 ± 0.7 and the 24 h AUC/MIC50 were 41.68 ± 0.1 and 42.26 ± 0.3 after administering the drug through PO and IV routes, respectively. The data in this study indicate that the application of 50 mg/kg b.w. of enrofloxacin to chicken through PO and IV routes with a dosing interval of 24 h can effectively cure S. Enteritidis infection, indicating the need for a 5-fold increase in the recommended dosage of enrofloxacin in chicken.


Assuntos
Antibacterianos/uso terapêutico , Enrofloxacina/uso terapêutico , Doenças das Aves Domésticas/tratamento farmacológico , Salmonelose Animal/tratamento farmacológico , Salmonella enteritidis/efeitos dos fármacos , Administração Oral , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacocinética , Galinhas/microbiologia , Enrofloxacina/administração & dosagem , Enrofloxacina/farmacocinética , Feminino , Injeções Intravenosas/veterinária , Masculino , Doenças das Aves Domésticas/microbiologia , Salmonelose Animal/microbiologia
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