RESUMEN
Previous studies have already demonstrated that mitochondria play a key role in Pb-induced apoptosis in primary cultures of rat proximal tubular (rPT) cells. To further clarify the underlying mechanism of Pb-induced mitochondrial apoptosis, this study was designed to investigate the role of mitochondrial permeability transition (MPT) and its regulatory components in Pb-induced apoptosis in rPT cells. Mitochondrial permeability transition pore (MPTP) opening together with disruption of mitochondrial ultrastructure, translocation of cytochrome c from mitochondria to cytoplasm and subsequent caspase-3 activation were observed in this study, suggesting that MPT is involved in Pb-induced apoptosis in rPT cells. Simultaneously, Pb-induced caspase-3 activation and apoptosis can be significantly inhibited by three MPTP inhibitors (CsA, DIDS, BA), which target different regulatory components of MPTP (Cyp-D, VDAC, ANT), respectively, demonstrating that Cyp-D, VDAC and ANT participate in MPTP regulation during lead exposure. Moreover, decreased ATP levels and increased ADP/ATP ratio induced by lead treatment can be significantly reversed by BA, indicating that Pb-mediated ANT dysfunction resulted in ATP depletion. In addition, up-regulation of VDAC-1, ANT-1 together with down-regulation of Cyp-D, VDAC-2 and ANT-2 at both the levels of transcription and translation were revealed in rPT cells under lead exposure conditions. In conclusion, Pb-mediated mitochondrial apoptosis in rPT cells is dependent on MPTP opening. Different expression levels in each isoform of three regulatory components contribute to alteration in their functions, which may promote the MPTP opening.
Asunto(s)
Apoptosis/efectos de los fármacos , Túbulos Renales Proximales/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Proteínas de Transporte de Membrana Mitocondrial/efectos de los fármacos , Compuestos Organometálicos/toxicidad , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Caspasa 3/metabolismo , Células Cultivadas , Citocromos c/metabolismo , Relación Dosis-Respuesta a Droga , Activación Enzimática , Regulación de la Expresión Génica , Túbulos Renales Proximales/metabolismo , Túbulos Renales Proximales/ultraestructura , Masculino , Mitocondrias/metabolismo , Mitocondrias/ultraestructura , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Poro de Transición de la Permeabilidad Mitocondrial , Compuestos Organometálicos/metabolismo , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacosRESUMEN
The aim of this study was to carry out a survey for the presence of Giardia duodenalis infection in canine using ELISA and PCR and to identify risk factors for infection. Samples from 318 dogs' feces living in nine cities in China were used in the present study. Each sample was tested for the presence of G. duodenalis-specific antigens using ELISA and 197 out of 318 samples were further examined for the presence of G. duodenalis using PCR. The overall rate of canines infected with giardiasis in the present study was 16.04% and 15.22% using ELISA and PCR, respectively. No significant difference was found between sex and Giardia positivity. Young dogs (up to one year) and living in communities were identified as risk factors for infection by multivariate logistic regression analysis. In conclusion, giardiasis in dogs was present in nine cities in China; as risk factors, young dogs (up to one year) and living in communities were of great significance. Giardia-infected canine should be treated for hygienic management to prevent transmission of the infection from dog to human.
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Enfermedades de los Perros/epidemiología , Giardia/aislamiento & purificación , Giardiasis/veterinaria , Animales , China/epidemiología , Enfermedades de los Perros/parasitología , Perros , Ensayo de Inmunoadsorción Enzimática/veterinaria , Heces/parasitología , Femenino , Giardiasis/epidemiología , Giardiasis/parasitología , Masculino , Reacción en Cadena de la Polimerasa/veterinaria , Prevalencia , Factores de RiesgoRESUMEN
Cadmium (Cd) is a toxic contaminant widely spread in natural and industrial environments. Adolescent exposure to Cd increases risk for obesity-related morbidity in young adults including type 2 diabetes and metabolic dysfunction-associated steatotic liver disease (MASLD). Despite this recognition, the direct impact of adolescent Cd exposure on the progression of MASLD later in life, and the mechanisms underlying these effects, remain unclear. Here, adolescent rats received control diet or diets containing 2 mg Cd2+/kg feed for 4 weeks, and then HFD containing 15% lard or control diet in young adult rats was selected for 6 weeks to clarify this issue. Data firstly showed that HFD-fed rats in young adulthood due to adolescent Cd exposure exhibited more severe MASLD, evidenced by increased liver damage, disordered serum and hepatic lipid levels, and activated NLRP3 inflammasome. Hepatic transcriptome analysis revealed the potential effects of mitochondrial dysfunction in aggravated MASLD due to Cd exposure. Verification data further confirmed that mitochondrial structure and function were targeted and disrupted during this process, shown by broken mitochondrial ridges, decreased mitochondrial membrane potential, imbalanced mitochondrial dynamic, insufficient ATP concentration, and enhanced mitochondrial ROS generation. However, mitophagy is inactively involved in clearance of damaged mitochondria induced by early Cd in HFD condition due to inhibited mitophagy receptor FUNDC1. In contrast, FUNDC1-dependent mitophagy activation prevents lipotoxicity aggravated by early Cd via suppressing mitochondrial ROS generation. Collectively, our data show that insufficient FUNDC1-dependent mitophagy can drive the transition from HFD-induced MASLD to MASH, and accordingly, these findings will provide a better understanding of potential mechanism of diet-induced metabolic diseases in the context of early environmental Cd exposure.
RESUMEN
A polymerase chain reaction (PCR) assay was performed in this study to amplify the major surface protein 5 (msp5) gene from the genomic DNA of Anaplasma marginale in Hyalomma asiaticum ticks by species-specific primers. Sequence analysis showed that the msp5 gene was 643 bases long and that the PCR products from the samples had an identical sequence (JX507127). Moreover, the BLAST showed that the sequence was identical to the msp5 sequences of A. marginale and most closely related to the A. marginale msp5 gene (AB704328) and the Liangdang strain of the A. marginale msp5 gene (EF546443) with similarity of 99 % (differing only by two bases). An epidemiological survey was performed in several dairy farms: a total of 68 ticks were collected from 49 cattle. As a result, 14 of the 49 (28.57 %) blood smears stained with Wright-Giemsa and 22 of the 68 (32.35 %) ticks examined by PCR assay exhibited A. marginale infection. The results of the PCR assay were mostly consistent with the results of the microscopic examination. A number of results were negative in blood smear but positive in PCR, which is important for the early diagnosis of anaplasmosis.
Asunto(s)
Anaplasma marginale/aislamiento & purificación , Técnicas Bacteriológicas/métodos , Entomología/métodos , Ixodidae/microbiología , Reacción en Cadena de la Polimerasa/métodos , Anaplasma marginale/genética , Animales , Proteínas Bacterianas/genética , Sangre/microbiología , Bovinos , Enfermedades de los Bovinos/parasitología , Cartilla de ADN/genética , Infestaciones Ectoparasitarias/parasitología , Infestaciones Ectoparasitarias/veterinaria , Homología de Secuencia de Ácido NucleicoRESUMEN
Babesiosis (piroplasmosis) is a zoonotic disease caused by an intraerythrocytic protozoan transmitted by Ixodes ticks. The aim of this study was to detect Babesia spp. infection using molecular methods in 377 blood samples from anemic patients. Sequence analysis showed that the 18S rRNA gene was 439 bases long by polymerase chain reaction (PCR) amplification and that the PCR products from the samples had an identical sequence (named Taian China, HM355854). BLAST search showed that the sequence was identical to the 18S rRNA sequences of Babesia divergens. The 18S rRNA sequence for Toxoplasma gondii was included as the outlier for phylogenetic analysis by using the program MEGA4.0 software. The results showed that the 18S rRNA gene sequences obtained from the present study was most closely related to B. divergens Switzerland (DQ312439) with 98.4% similarity (differing only by seven bases). The phylogenetic analysis also revealed that this sequence closely resembled B. divergens strains from other countries and belonged to the same clade. This is the first report of a human being infected by B. divergens in China.
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Anemia/diagnóstico , Anemia/parasitología , Babesia/aislamiento & purificación , Babesiosis/complicaciones , Babesiosis/diagnóstico , Adolescente , Adulto , Anciano , Babesia/clasificación , Babesiosis/patología , Niño , Preescolar , China , Análisis por Conglomerados , ADN Protozoario/química , ADN Protozoario/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Femenino , Genes de ARNr , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , ARN Protozoario/genética , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Adulto JovenRESUMEN
Previous studies have shown that oxidative stress-induced inhibition of autophagy plays a pivotal role in cadmium (Cd)-mediated cytotoxicity in primary rat proximal tubular (rPT) cells. The objective of this study is to explore the protective effect of puerarin (PU), a potent antioxidant, on Cd-induced autophagy inhibition and oxidative stress in rPT cells. First, Cd-induced blockage of autophagic flux in rPT cells was obviously restored by PU treatment, evidenced by immunoblot analysis of autophagy marker proteins and tandem fluorescent-tagged LC3 method. Resultantly, Cd-induced autophagosome accumulation was significantly alleviated by PU treatment. Also, Cd-induced lysosomal alkalinization and impairment of lysosomal degradation capacity were obviously recovered by PU, demonstrating that PU can restore Cd-induced lysosomal dysfunction. Moreover, Cd-induced lysosomal membrane permeabilization (LMP) was effectively blocked by PU. Cd-stimulated Nrf2 nuclear translocation and subsequent elevated expression of Nrf2-downstream targets were significantly inhibited by PU treatment. Simultaneously, Cd-elevated protein levels of antioxidant enzymes and glutathione synthesis-related proteins in rPT cells were markedly downregulated by PU treatment. In conclusion, these observations indicate that PU alleviates Cd-induced cytotoxicity in rPT cells through restoring autophagy, blocking LMP and inhibiting Nrf2 pathway, which is intimately related with its antioxidant activity.
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Cadmio/toxicidad , Isoflavonas/farmacología , Túbulos Renales Proximales/efectos de los fármacos , Lisosomas/efectos de los fármacos , Factor 2 Relacionado con NF-E2/antagonistas & inhibidores , Vasodilatadores/farmacología , Animales , Células Cultivadas , Relación Dosis-Respuesta a Droga , Túbulos Renales Proximales/metabolismo , Lisosomas/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiologíaRESUMEN
Lead (Pb) is a known nephrotoxicant that causes damage to proximal tubular cells. PERK pathway plays an important role in the pathogenesis of renal diseases, but its role in Pb-induced nephrotoxicity remains largely unknown. In this study, data showed that Pb could induce ER stress as shown by increased phosphorylation of PERK with subsequent activation of the eIF2α-ATF4-CHOP axis in primary rat proximal tubular (rPT) cells, indicating the activation of PERK-eIF2α-ATF4-CHOP pathway due to excessive ER stress. Pb-activated PERK pathway can be effectively inhibited by 4-phenylbutyric acid and PERK gene silencing, respectively; whereas continuously up-regulated by tunicamycin (TM) treatment. Moreover, Pb-induced apoptosis and inhibition of autophagic flux in rPT cells were significantly augmented and aggravated by co-treatment with TM, respectively. Pharmacological or genetic inhibition of the PERK pathway results in alleviation of apoptosis and restoration of autophagy inhibition in Pb-exposed rPT cells. Mechanistically, activation of PERK-eIF2α-ATF4-CHOP axis triggered by excessive ER stress in rPT cells leads to Pb-induced apoptosis and blockage of autophagic flux, resulting in nephrotoxicity.
Asunto(s)
Estrés del Retículo Endoplásmico , Túbulos Renales Proximales/efectos de los fármacos , Plomo/toxicidad , eIF-2 Quinasa/metabolismo , Factor de Transcripción Activador 4/metabolismo , Animales , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Células Cultivadas , Estrés del Retículo Endoplásmico/efectos de los fármacos , Factor 2 Eucariótico de Iniciación/metabolismo , Túbulos Renales Proximales/enzimología , Túbulos Renales Proximales/metabolismo , Fenilbutiratos/farmacología , Ratas Sprague-Dawley , Factor de Transcripción CHOP/metabolismo , Tunicamicina/farmacología , eIF-2 Quinasa/genéticaRESUMEN
Autophagy and apoptosis are two different biological processes that determine cell fates. We previously reported that autophagy inhibition and apoptosis induction are involved in lead(II)-induced cytotoxicity in primary rat proximal tubular (rPT) cells, but the interplay between them remains to be elucidated. Firstly, data showed that lead(II)-induced elevation of LC3-II protein levels can be significantly modulated by 3-methyladenine or rapamycin; moreover, protein levels of Autophagy-related protein 5 (Atg5) and Beclin-1 were markedly up-regulated by lead(II) treatment, demonstrating that lead(II) could promote the autophagosomes formation in rPT cells. Next, we applied three pharmacological agents and genetic method targeting the early stage of autophagy to validate that enhancement of autophagosomes formation can inhibit lead(II)-induced apoptotic cell death in rPT cells. Simultaneously, lead(II) inhibited the autophagic degradation of rPT cells, while the addition of autophagic degradation inhibitor bafilomycin A1 aggravated lead(II)-induced apoptotic death in rPT cells. Collectively, this study provided us a good model to know about the dynamic process of lead(II)-induced autophagy in rPT cells, and the interplay between autophagy and apoptosis highlights a new sight into the mechanism of lead(II)-induced nephrotoxicity.
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Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Túbulos Renales Proximales/citología , Plomo/toxicidad , Adenina/análogos & derivados , Adenina/farmacología , Animales , Proteína 5 Relacionada con la Autofagia/metabolismo , Beclina-1/metabolismo , Células Cultivadas , Proteínas Asociadas a Microtúbulos/metabolismo , Ratas , Sirolimus/farmacologíaRESUMEN
Lead (Pb) is a known nephrotoxic element. Recently we have proved that subcellular Ca2+ redistribution is involved in Pb-induced apoptosis in primary cultures of rat proximal tubular (rPT) cells, but the underlying mechanism remains to be elucidated. Firstly, data showed that Pb triggers endoplasmic reticulum (ER) stress response in rPT cells, as evidenced by the elevations of ER stress markers. Moreover, pharmacological modulation of Ca2+ mobilization in ER and cytoplasm with three chemicals (2-APB or TG or BAPTA-AM) can effectively increase or decrease the protein expression of ER stress markers in Pb-exposed rPT cells, demonstrating that Pb-induced ER stress is Ca2+-dependent. We found that Pb stimulates phosphorylation of calcium/calmodulin-dependent protein kinase II (CaMKII) to activate its activity. Meanwhile, inhibition of CaMKII with KN93 or KN62 attenuated Pb-activated caspase-12 and CCAAT/enhancer-binding protein homologous protein (CHOP) in rPT cells, demonstrating that CaMKII activation promoted ER stress in rPT cells. Likewise, Pb-induced apoptosis can be effectively inhibited by CaMKII inhibitor KN93 or KN62. Furthermore, co-treatment with KN93 or KN62 significantly reversed Pb-induced ER Ca2+ release and concomitant intracellular Ca2+ overload in rPT cells. In summary, these results expound the mechanisms involving in ER stress, Ca2+ dyshomeostasis and activated CaMKII, which all contribute to Pb-induced apoptosis. CaMKII acts as a critical mediator of ER stress and associated apoptosis via regulating intracellular Ca2+ mobilization from ER to cytoplasm.
RESUMEN
Autophagy has an important renoprotective function and we recently found that autophagy inhibition is involved in cadmium (Cd)-induced nephrotoxicity. Here, we aimed to investigate the protective effect of trehalose (Tre), a novel autophagy activator, against Cd-induced cytotoxicity in primary rat proximal tubular (rPT) cells. First, data showed that Tre treatment significantly decreased Cd-induced apoptotic cell death of rPT cells via inhibiting caspase-dependent apoptotic pathway, evidenced by morphological analysis, flow cytometric and immunoblot assays. Also, administration with Tre protected rPT cells against Cd-induced lipid peroxidation. Inhibition of autophagic flux in Cd-exposed rPT cells was markedly restored by Tre administration, demonstrated by immunoblot analysis of autophagy marker proteins and GFP and RFP tandemly tagged LC3 method. Resultantly, Cd-induced autophagosome accumulation was obviously alleviated by Tre treatment. Meanwhile, blockage of autophagosome-lysosome fusion by Cd exposure was noticeably restored by Tre, which promoted the autophagic degradation in Cd-exposed rPT cells. Moreover, Tre treatment markedly recovered Cd-induced lysosomal alkalinization and impairment of lysosomal degradation capacity in rPT cells, demonstrating that Tre has the ability to restore Cd-impaired lysosomal function. Collectively, these findings demonstrate that Tre treatment alleviates Cd-induced cytotoxicity in rPT cells by inhibiting apoptosis and restoring autophagic flux.
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Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Cadmio/toxicidad , Túbulos Renales Proximales/patología , Sustancias Protectoras/farmacología , Trehalosa/farmacología , Animales , Autofagosomas/fisiología , Caspasa 3/metabolismo , Caspasa 9/metabolismo , Células Cultivadas , Túbulos Renales Proximales/citología , Túbulos Renales Proximales/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Lisosomas/fisiología , Estrés Oxidativo/efectos de los fármacos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Cultivo Primario de Células , Ratas , Ratas Sprague-DawleyRESUMEN
Previous study has demonstrated that mitochondrial-dependent apoptotic pathway is involved in the nephroprotective effect of puerarin (PU) against lead-induced cytotoxicity in primary cultures of rat proximal tubular (rPT) cells. To further clarify how PU exerts its antiapoptotic effects, this study was designed to investigate the role of mitochondrial permeability transition (MPT) and subsequent apoptotic events in the process of PU against Pb-induced cytotoxicity in rPT cells. The results showed that Pb-mediated mitochondrial permeability transition pore (MPTP) opening together with mitochondrial cytochrome c release, activations of caspase-9 and caspase-3, and subsequent poly-ADP-ribose polymerase (PARP) cleavage can be effectively blocked by the addition of PU. Simultaneously, upregulation and downregulation of Bcl-2 and Bax with increased Bcl-2/Bax ratio due to PU administration further alleviated Pb-induced mitochondrial apoptosis. Moreover, PU can reverse Pb-induced ATP depletion by restoring mitochondrial fragmentation to affect ATP production and by regulating expression levels of ANT-1 and ANT-2 to improve ATP transport. In summary, PU produced a significant protection against Pb-induced mitochondrial apoptosis in rPT cells by inhibiting MPTP opening to ameliorate the mitochondrial dysfunction.
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Apoptosis/efectos de los fármacos , Isoflavonas/farmacología , Túbulos Renales Proximales/metabolismo , Plomo/toxicidad , Mitocondrias/metabolismo , Proteínas de Transporte de Membrana Mitocondrial , Animales , Células Cultivadas , Proteínas de Transporte de Membrana Mitocondrial/antagonistas & inhibidores , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Poro de Transición de la Permeabilidad Mitocondrial , Cultivo Primario de Células , RatasRESUMEN
Previous studies have shown that cytosolic Ca(2+) ([Ca(2+)]c) overload was involved in Pb-induced apoptosis in primary cultures of rat proximal tubular (rPT) cells, but the source of elevated Ca(2+) and the effect of potential subcellular Ca(2+) redistribution on apoptosis are still unknown. In this study, variations of [Ca(2+)]c in two culture media (Ca(2+)-containing and Ca(2+)- free) were analyzed, indicating that Pb-induced elevation of [Ca(2+)]c was primarily generated intracellularly. Fluo-4-AM, dihydro-Rhod-2-AM and Mag-Fluo-4-AM was loaded to Pb-exposed rPT cells to monitor the imaging of Ca(2+) concentrations in the cytoplasm ([Ca(2+)]c), mitochondria ([Ca(2+)]mit) and endoplasmic reticulum (ER) ([Ca(2+)]ER), respectively, under the confocal microscope. Data indicate that elevations of [Ca(2+)]c and [Ca(2+)]mit with depletion of [Ca(2+)]ER were revealed in Pb-treated rPT cells, but this subcellular Ca(2+) redistribution could be significantly suppressed by 2-APB, a specific inhibitor of inositol 1,4,5-trisphosphate receptor (IP3R) that functions to release Ca(2+) from ER stores. Simultaneously, Pb-mediated mitochondrial Ca(2+) overload can be partially suppressed by the cytosolic Ca(2+) chelator BAPTA-AM, suggesting that Ca(2+) uptake into mitochondria occurs via diverse pathways and ER Ca(2+) storage was the chief source. Furthermore, Pb-induced apoptosis was markedly inhibited by 2-APB and BAPTA-AM, respectively. Additionally, elevated IP3 levels with up-regulated IP3R-1 and IP3R-2 (mRNA and protein) levels were revealed in Pb-exposed rPT cells. In summary, IP3R-mediated ER Ca(2+) release promoted the elevations of [Ca(2+)]c and [Ca(2+)]mit in Pb-exposed rPT cells, which played a chief role in apoptosis induced by impaired calcium homeostasis.
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Apoptosis/efectos de los fármacos , Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Túbulos Renales Proximales/efectos de los fármacos , Compuestos Organometálicos/toxicidad , Animales , Compuestos de Boro/farmacología , Células Cultivadas , Quelantes/farmacología , Citoplasma/metabolismo , Relación Dosis-Respuesta a Droga , Ácido Egtácico/análogos & derivados , Ácido Egtácico/farmacología , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Homeostasis , Inositol 1,4,5-Trifosfato/metabolismo , Receptores de Inositol 1,4,5-Trifosfato/efectos de los fármacos , Receptores de Inositol 1,4,5-Trifosfato/metabolismo , Túbulos Renales Proximales/metabolismo , Túbulos Renales Proximales/patología , Microscopía Confocal , Microscopía Fluorescente , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Cultivo Primario de Células , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
A one-step immunochromatographic assay using gold nanoparticles coated with polyclonal antibody (pAb) against Mycoplasma suis (M. suis) was developed in this study for the detection of M. suis in porcine plasma. The colloidal gold was prepared by the reduction of gold salt with sodium citrate coupled with pAb against M. suis. The pAb was produced by immunizing the BALB/c mice with recombinant MSG1 (rMSG1) protein from M. suis expressed in Escherichia coli. The optimal concentrations of the capture antibody and the coating antibody were 12 µg/ml and 1.5 mg/ml, respectively, and that of the blocking buffer was 1% bovine serum albumin. The lower detection limit of the immunochromatographic assay test was 100 ng/ml with visual detection under optimal conditions of analysis. Classical swine fever virus, porcine reproductive and respiratory syndrome virus, swine pneumonia mycoplasma, swine toxoplasma, and porcine parvovirus were used to evaluate the specificity of the immunochromatographic strips. No cross-reaction of the antibodies with other related swine pathogens was observed. This qualitative test based on the visual evaluation of the results did not require any equipment. The assay time for M. suis detection was less than 10 min, suitable for rapid detection at the grassroots level. The one-step colloidal gold immunochromatographic strips that we developed had high specificity and sensitivity. Therefore, this method would be feasible, convenient, rapid, and effective for detecting M. suis in porcine plasma.
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Anticuerpos/inmunología , Antígenos Bacterianos/sangre , Cromatografía de Afinidad/instrumentación , Oro/química , Mycoplasma/inmunología , Mycoplasma/aislamiento & purificación , Neumonía Porcina por Mycoplasma/sangre , Animales , Antígenos Bacterianos/inmunología , Técnicas Biosensibles/instrumentación , Coloides/química , Diseño de Equipo , Análisis de Falla de Equipo , Nanopartículas del Metal/química , Ratones , Ratones Endogámicos BALB C , Neumonía Porcina por Mycoplasma/diagnóstico , Neumonía Porcina por Mycoplasma/inmunología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , PorcinosRESUMEN
Puerarin (PU), a natural flavonoid, has been reported to possess anti-oxidative and anti-inflammatory activities. In the present study, female Sprague-Dawley rats received lead (Pb) nitrate (300 mg/L, via drinking water) and/or PU (400 mg/kg/day, orally) to investigate the protective effects of PU on Pb-induced renal damage. Renal toxicity was evaluated by detecting urinary proteins excretion as well as levels of serum urea nitrogen and serum creatinine. Ultrastructural observations and real-time quantitative polymerase chain reaction analyses were performed on kidney cortex tissues to identify the mitochondrial damage and quantify gene expression levels of cytochrome oxidase submits (COX-I/II/III), respectively. Renal cell damage was assessed by light microscopic examination. Lipid peroxidation (LPO) levels and antioxidant status in kidney were also evaluated. Animals that received both Pb and PU showed a better renal function than those that received Pb alone, with minor pathological damage. Moreover, PU significantly reduced LPO and markedly restored the enzymatic and non-enzymatic antioxidants levels in kidney of Pb-treated rats, which may be related to its restoring mitochondrial function. Furthermore, PU administration significantly increased urinary Pb excretion and decreased its level in the serum and kidney. In conclusion, these results suggested that PU reduces renal damage induced by chronic Pb administration through its antioxidant properties and chelating ability.
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Isoflavonas/uso terapéutico , Enfermedades Renales/tratamiento farmacológico , Intoxicación por Plomo/tratamiento farmacológico , Sustancias Protectoras/uso terapéutico , Animales , Creatinina/sangre , Complejo IV de Transporte de Electrones/genética , Femenino , Isoflavonas/farmacología , Enfermedades Renales/inducido químicamente , Enfermedades Renales/metabolismo , Enfermedades Renales/patología , Plomo/sangre , Plomo/farmacocinética , Plomo/toxicidad , Plomo/orina , Intoxicación por Plomo/metabolismo , Intoxicación por Plomo/patología , Sustancias Protectoras/farmacología , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Urea/sangreRESUMEN
Seroprevalence of Toxoplasma gondii infection was determined in sera of 632 dogs (551 pets, 81 strays) from Shandong, Henan, and Heilongjiang Provinces, and in the Xinjiang Uygur Autonomous Region, People's Republic of China (PRC), using the indirect hemagglutination assay (cutoff titer 1:64 or higher); 11.1% were seropositive. The seroprevalence in stray dogs and in ≥3-yr-old dogs was significantly higher (P < 0.05) than that in household dogs and in <3-yr-old dogs. There were no significant differences in terms of gender, breed, or locality (P ≥ 0.05). The results indicate that T. gondii infections are common in dogs in PRC.
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Anticuerpos Antiprotozoarios/sangre , Enfermedades de los Perros/epidemiología , Toxoplasma/inmunología , Toxoplasmosis Animal/epidemiología , Distribución por Edad , Animales , China/epidemiología , Enfermedades de los Perros/parasitología , Perros , Femenino , Pruebas de Hemaglutinación/veterinaria , Masculino , Estudios Seroepidemiológicos , Distribución por SexoRESUMEN
The recombinant ppa protein of Mycoplasma suis migrated to 21 kDa. Using this antigen, an ELISA system to detect the antibody against M. suis infection in swine was established. The rELISA demonstrated 98.5% specificities among negative samples and 96.9% sensitivity among positive samples with M. suis infection. A comparison of this ELISA system with an indirect hemagglutination assay (IHA) test using 132 swine samples revealed that the positive rate was 34.0% in ELISA and 28.0% in IHA. Compared with IHA, the present rELISA system using recombinant ppa antigen significantly improves the specificity, sensitivity, and stability for serodiagnosis of M. suis infection in swine.
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Anticuerpos Antibacterianos/inmunología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Pirofosfatasa Inorgánica/inmunología , Infecciones por Mycoplasma/veterinaria , Mycoplasma/inmunología , Enfermedades de los Porcinos/diagnóstico , Animales , Antígenos Bacterianos/genética , Antígenos Bacterianos/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Pruebas de Hemaglutinación/veterinaria , Pirofosfatasa Inorgánica/genética , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/inmunología , Proteínas Recombinantes/inmunología , Porcinos , Enfermedades de los Porcinos/inmunologíaRESUMEN
The present study conducts a serological survey on the presence of canine dirofilariosis in domestic dogs using an enzyme-linked immunosorbent assay (ELISA) kit. A total of 310 household dogs (166 females and 144 males) in Chongqing, Kunming, Nanchang, Fuzhou, Guangzhou, Shenzhen, and Nanning in Southern China were examined. Of the 310 dogs, 42 (13.5%) were seropositive for dirofilariosis. No statistically significant difference was observed in terms of sex in the seroprevalence of dirofilariosis using the ELISA kit. The positive rates for dirofilariosis were 6.6% in the 0-1-year-old group, 13.8% in the 1-4-year-old group, and 21.6% in the less than 4-year-old group. The statistical analysis revealed that significant differences were observed in the 1-4-year-old group (P=0.037, OR=0.441, 95% CI=0.170-1.144) and less than 4-year-old group (P<0.001, OR=0.256, 95% CI=0.095-0.693). In the regional comparison, the shoreline city Shenzhen (18.8%) had a significantly higher prevalence than urban and mountain areas (P<0.05, OR=0.310, 95% CI=0.066-1.445). In conclusion, Dirofilaria immitis infection in domestic dogs was present in Chongqing, Kunming, Nanchang, Fuzhou, Guangzhou, Shenzhen, and Nanning. Therefore, heartworm treatment and/or chemoprophylaxis for the captured domestic dogs are necessary in these areas. To the best of our knowledge, the present study is the first using serological methods to examine D. immitis infection in domestic dogs in Mainland China in the recent years.
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Dirofilariasis/sangre , Enfermedades de los Perros/sangre , Animales , China/epidemiología , Dirofilariasis/epidemiología , Enfermedades de los Perros/epidemiología , Perros , Femenino , Masculino , Estudios SeroepidemiológicosRESUMEN
Lead acetate (300 mg/L) and/or cadmium chloride (50 mg/L) were administered as drinking water to Sprague-Dawley rats for 9 weeks to investigate the effects of concurrent exposure to lead and cadmium on the distribution patterns of five essential trace elements. Inductively coupled plasma mass spectrometry was used to determine the concentrations of zinc, copper, manganese, selenium and iron in the urine at different exposure times, as well as their levels in the renal cortex and serum at the end of treatment. Compared with the control group, exposure to lead and/or cadmium resulted in a significant increase in the urinary excretion of these five elements during the experiment, whereas significant decreased levels of these elements were found in kidney and serum. In conclusion, increased urinary loss of antioxidant trace elements due to lead and/or cadmium exposure induced the deficiency of antioxidants in the body, which could result in further oxidative damage. Moreover, there was an obvious synergistic effect of lead combined with cadmium on the distribution patterns of these essential trace elements, which may be related to the severity of co-exposure to these two metals.
Asunto(s)
Cloruro de Cadmio/farmacocinética , Contaminantes Ambientales/farmacología , Compuestos Organometálicos/farmacocinética , Oligoelementos/farmacocinética , Animales , Antioxidantes/farmacocinética , Cloruro de Cadmio/toxicidad , Agua Potable , Sinergismo Farmacológico , Contaminantes Ambientales/toxicidad , Femenino , Corteza Renal/química , Corteza Renal/efectos de los fármacos , Corteza Renal/metabolismo , Compuestos Organometálicos/toxicidad , Ratas , Ratas Sprague-Dawley , Espectrofotometría Atómica , Oligoelementos/análisis , Oligoelementos/orinaRESUMEN
Mycoplasma suis (M. suis), a hemotrophic pathogen of pigs, causes economic losses in swine production throughout the world. Inorganic pyrophosphatase (ppa) is a very important gene in M. suis. The ppa gene of M. suis was synthesized by PCR-based accurate synthesis (PAS) and overlapextension PCR, inserted into vector pMD18-T, and then subcloned to the prokaryotic expression vector pET28c.The recombinant plasmid pET28c_ppa was transformed to E. coli BL21 for expression under induction of isopropyl thiogalactoside. The expressed product was identified by SDS-PAGE and Western blot, which suggested that the recombinant protein has good antigenicity. Piglets were immunised with purified recombinant protein, and specific antibodies to the recombinant protein were detected in piglet serum. The results show that the ppa gene can be efficiently expressed in E. coli and that the expressed recombinant protein can elicit a specific serum antibody response in piglets. PAS and overlap-extension PCR were first used to synthesize the ppa of M. suis. They provide simple, rapid, reliable and relatively inexpensive methods to synthesize, clone, and express genes. The experiment conducted in this paper will enable future research into the role and function of the ppa gene.