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1.
ACS Omega ; 7(27): 23875-23889, 2022 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-35847299

RESUMO

Nanoencapsulation with safe materials improves delivery, stability, and activity of bioactive components. We report a novel safe, and effective method for the development of encapsulated antimicrobial essential oils (EO) for topical creams and gels. The method developed features three aspects that, to our knowledge, had not been previously demonstrated: (1) use of novel liposomes (LPs) to encapsulate EOs, (2) use of the EOs to replace synthetic organic solvents that are potentially toxic and/or leave harmful residues, and (3) an encapsulation process at temperatures below the boiling point of water. The LPs were made from soy lecithin, phytosterol, and α-tocopherol (vitamin E) that were synthesized using the EOs as the solvent. The liposomes were converted to nanoliposomes (NLPs) through a series of sonication, homogenization, and extrusion steps. Transmission electron microscopy indicated that the NLPs alone and nanoliposome encapsulated EOs (NLP-EOs) were spherical in shape with sizes ranging between 50 and 115 nm diameter and with negative zeta potentials ranging from -34 to -43 mV. There was no significant heavy metal contamination [As, Pb, Cd, Hg] based on inductively coupled plasma (ICP) mass spectrometry MS analyses. Nearly complete EO encapsulation (95% encapsulation efficiency) was achieved and confirmed by GC/MS. Three of the NLP-EOs made of various essential oils were used to make topical formulations (cream and gel) which exhibited antimicrobial activities against Escherichia coli (Gram negative) and Bacillus subtilis (Gram positive) bacteria. The creams with NLP-EOs were as active against the two bacteria in the antimicrobial assays as the conventional antibiotic Kanamycin that was used as positive control.

2.
Anal Biochem ; 646: 114647, 2022 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-35289287

RESUMO

Salmonella infection could come from eating contaminated meat or raw eggs, and drinking milk or water contaminated by Salmonella enteritidis. Therefore, it is necessary to explore a fast and easy method for the detection of S. enteritidis in these diverse samples. For this purpose, a novel particle size sensing tool was designed for ultrasensitive and accurate S. enteritidis detection. This assay consisted of rolling circle amplification (RCA) with dynamic light scattering (DLS) using gold nanoparticles (AuNPs) modified with DNA probe as DNA-AuNPs as the capture surface into a hybrid RCA-DLS assay combined with asymmetric polymerase chain reaction (aPCR) and subsequent detection. Under optimal experimental conditions, the novel hybrid RCA-DLS assay combined with aPCR for S. enteritidis reached a limit of detection (LOD) as low as 3 × 100 CFU/mL in pure culture. In spiked milk samples, the LOD was 2.0 × 100 CFU/mL without pre-enriched bacteria. The total time of RCA-DLS assay was about 6 h which including genomic DNA extraction, aPCR, RCA and DLS determination. The hybrid RCA-DLS assay combined with aPCR holds promise in the specific and sensitive S. enteritidis detection.


Assuntos
Nanopartículas Metálicas , Salmonella enteritidis , DNA Bacteriano/análise , DNA Bacteriano/genética , Difusão Dinâmica da Luz , Ouro , Limite de Detecção , Reação em Cadeia da Polimerase/métodos , Salmonella enteritidis/genética
3.
Toxicol Res (Camb) ; 10(4): 937-946, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34484685

RESUMO

Titanium dioxide nanoparticles (TiO2 NPs) have been widely used in food, medical, and other fields; their reproductive toxicity has been reported in numerous studies. However, the relevant toxicity mechanism still requires further exploration. In this paper, the effect of oral exposure to 500 mg/kg TiO2 NPs (anatase and rutile) in adult male SD rats was studied over 3 and 7 days. Results showed that the total sperm count and testosterone level of 7 days of exposure in serum decreased in the experimental group. Testicular tissue lesions, such as disappearance of Leydig cells, disorder of arrangement of spermatogenic cells in the lumen of convoluted seminiferous tubules, and disorder of arrangement of germ cells, were observed. Meanwhile, the expression of steroidogenic acute regulatory (StAR; the key factors of testosterone synthesis), MAPK (ERK1/2), and phosphorylated ERK1/2 in testes of SD rats after exposure to TiO2 NPs for 7 days decreased, while the malondialdehyde content increased and superoxide dismutase activity decreased in serum. The present study showed that TiO2 NPs could cause reproductive toxicity. Notably, anatase is more toxic than rutile. In addition, exposure to 500 mg/kg TiO2 NPs for 7 days inhibited testosterone synthesis in male rat, which may be related to the reactive oxygen species (ROS)-MAPK (ERK1/2)-StAR signal pathway. Warning that the use of TiO2 NPs should be regulated.

4.
J Dairy Sci ; 104(12): 12295-12302, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34538487

RESUMO

Salmonella is a foodborne pathogen that has contributed to numerous food safety accidents worldwide, making it necessary to detect contamination at an early stage. A pair of specific primers based on the invA gene of Salmonella was designed for PCR. Target double-stranded DNA (dsDNA) from PCR was purified and denatured at high temperature to obtain target single-stranded DNA (ssDNA). Two carboxyfluorescein-labeled hairpin probes (H1-FAM and H2-FAM) were designed with complementary portions to the ssDNA sequence so that binding could trigger H1-FAM and H2-FAM hybridization chain reaction (HCR) to produce a long dsDNA complex. In this study, graphene oxide (GO) was used in the development of a homogeneous fluorescence detection platform for Salmonella. Using this HCR-GO assay platform, Salmonella detection was completed in 3.5 h. Salmonella was reliably and specifically detected with a limit of detection (LOD) of 4.2 × 101 cfu/mL in pure culture. Moreover, this new HCR-GO assay platform was successfully applied to the detection of Salmonella in artificially contaminated milk with a LOD of 4.2 × 102 cfu/mL.


Assuntos
Técnicas Biossensoriais , Grafite , Animais , Técnicas Biossensoriais/veterinária , Leite , Salmonella/genética
5.
Environ Toxicol ; 36(6): 1143-1151, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33599401

RESUMO

The potential toxicity of Zinc oxide nanoparticles (ZnO NPs) to human beings has become a widespread concern. This study explored the reproductive toxicity and the mechanism of toxicity of ZnO NPs in early pregnant mice. The results showed that abnormal weight changes, induced inflammation, reduced level of serum sex hormones, damaged uterus, increased abortion, and abnormal development of fetus. In the uterus, the transcription levels of ZnT-1, HO-1, Bax, Bax/Bcl-2, JNK, and Caspase-3 were significantly up-regulated while Bcl-2, ER-1 and PR were significantly down-regulated. The TUNEL-positive cells increased that were exposed to high levels of ZnO NPs. In summary, those results indicated that Zn from high levels of exposure to ZnO NPs accumulated in the uterus that could have caused the formation of ROS that led to oxidative stress, which might have activated the mitochondrial apoptotic pathway that could have caused the uterine injury which induced the observed reproductive toxicity.


Assuntos
Nanopartículas , Óxido de Zinco , Animais , Apoptose , Feminino , Camundongos , Mitocôndrias , Estresse Oxidativo , Gravidez , Espécies Reativas de Oxigênio , Óxido de Zinco/toxicidade
6.
Talanta ; 225: 121953, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33592708

RESUMO

Sepsis caused by bacteria has high morbidity and mortality, and it is neccerssay to establish a fast, convenient, and facility assays for detection of bacteria. In this study, we have developed established a simple, rapid, and ultrasensitive vancomycin (Van) and dendrimer nanoparticles-based method to isolate and detect bacteria in human blood using a multivalent binding strategy. The proposed Bio-den-Van multivalent capture nanoplatform combined with m-qPCR for simultaneous detection of two kinds of bacteria was demonstrated with rapid 2 min bacteria isolation with a linear range at 3.2 × 101-3.2 × 106 CFU·mL-1 for L. monocytogenes and 4.1 × 101-4.1 × 106 CFU·mL-1 for S. aureus, respectively. The limit of detection (LOD) for simultaneous detection of L. monocytogenes and S. aureus were 32 and 41 CFU·mL-1 in spiked human blood samples, respectively. Other bacteria had an insignificant interference with the test results. This Bio-den-Van multivalent capture nanoplatform combined with m-qPCR detection exhibited rapid, high sensitivity and specificity in simultaneous detection of various bacteria. To our knowledge, this is the first time that Bio-den-Van multivalent capture nanoplatform was used with Van as a recognition molecule for the simultaneous capture and subsequent detection of two bacteria from spiked human blood sample. This method holds great potential for future clinical applications.


Assuntos
Dendrímeros , Vancomicina , Bactérias , Humanos , Fenômenos Magnéticos , Sensibilidade e Especificidade , Staphylococcus aureus/genética , Vancomicina/farmacologia
7.
J Dairy Sci ; 104(2): 1465-1473, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33272582

RESUMO

In this study, a new vancomycin (Van)-modified poly-l-lysine (PLL) magnetic bead (MB) technique was developed for isolation of gram-positive bacteria. The method combines magnetic separation with a multiplex PCR (mPCR) assay and gel electrophoresis for easy and rapid detection of Bacillus cereus. Vancomycin was used as a molecular ligand between the MB and the d-alanyl-d-alanine moieties on the cell wall surface of B. cereus. The PLL served as a flexible molecular tether between the MB and Van that reduced steric hindrance maintaining the biological activity of Van. The MB-PLL-Van capture nanoprobes exhibited excellent capture and isolation efficiency for B. cereus in spiked milk matrix samples without interference from the complex food matrix. The subsequent mPCR assay showed high specificity for the 4 target genes in B. cereus, the entFM, cesB, cer, and 16S rRNA genes, that were used to achieve efficient genotyping and detection. Under optimum conditions, the limit of detection reached 103 cfu/mL, with a dynamic range of detection at 103 to 107 cfu/mL in pure culture. Application of the MB-PLL-Van mediated mPCR assay for B. cereus in milk matrix samples achieved results similar to those of the pure culture. In addition, with a 6-h pre-enrichment of B. cereus that was spiked in milk matrix samples, the limit of detection reached 101 cfu/mL. The MB-PLL-Van mediated mPCR assay developed in this study could be used as a universal technology platform for the efficient enrichment and genotyping of gram-positive bacteria.


Assuntos
Bacillus cereus/isolamento & purificação , Campos Magnéticos , Leite/microbiologia , Reação em Cadeia da Polimerase Multiplex , Polilisina , Vancomicina , Animais , Bacillus cereus/classificação , Bacillus cereus/genética , Microbiologia de Alimentos/métodos , Técnicas de Genotipagem , Microesferas , Reação em Cadeia da Polimerase Multiplex/veterinária , RNA Ribossômico 16S
8.
Environ Toxicol ; 36(4): 550-561, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33236829

RESUMO

ZnO nanoparticles (NPs) are among the most manufactured nanoparticles in the consumer products, industries, and researches. An increasing body of evidence indicated that ZnO NPs show toxicological effects in vivo. Sex differences in the toxicity of ZnO NPs are not clear, thus the aim of this study was to investigate the effects of ZnO NPs on the female and male reproductive organs (uterus, ovary and testes). ZnO NPs were orally administered to female and male mice at dosages level of 0 and 100 mg/kg body weight. The biological material was sampled 3 days after tube feeding. The results demonstrated that Zinc contents were accumulated in the reproductive organs of treated mice. Furthermore, ZnO NPs administration induced significant decrease in the testes weight, an imbalance of hematological and serum biochemical parameters in male mice. The histopathological examinations showed that structural disorder and the appearance of cell apoptosis and death in the ZnO NPs-exposed mice. Additionally, the RT-qPCR data indicated ZnO NPs can activate mitochondrial-mediated signaling pathway and induce caspase depend damage that ultimately injured the uterus. In the ovary, ZnO NPs induce cell apoptosis in Shh pathway activated ovary cells, and affect the synthesis of steroidogenesis. In the testes, ZnO NPs effectively changed the expression level of genes related to oxidant stress, detox/metabolic process, and apoptosis. It was found that ZnO NPs caused more serious reproductive toxicity in the male mice than female mice. Overall, these findings indicated that ZnO NPs could induce exposure-related risks to reproductive health, especially in those who are at the occupational level.


Assuntos
Expressão Gênica/efeitos dos fármacos , Nanopartículas/toxicidade , Ovário/efeitos dos fármacos , Testículo/efeitos dos fármacos , Útero/efeitos dos fármacos , Óxido de Zinco/toxicidade , Administração Oral , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Bioacumulação , Biomarcadores/sangue , Caspases/genética , Feminino , Masculino , Camundongos , Nanopartículas/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Ovário/metabolismo , Ovário/patologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Transdução de Sinais , Testículo/metabolismo , Testículo/patologia , Útero/metabolismo , Útero/patologia , Zinco , Óxido de Zinco/metabolismo
9.
Ecotoxicol Environ Saf ; 206: 111393, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33010597

RESUMO

Human were given a lot of opportunities to ingest TiO2 NPs in the environment. Children have low, sensitive intestinal tolerance, and they could be exposed to higher levels of TiO2 NPs than adults. Few studies have been conducted on the interaction between TiO2 NPs and juvenile intestine phase models. Thus, in this work, weaning rats were orally exposed to TiO2 NPs for 7 and 14 days. Results indicate that Ti accumulated in the intestine, liver, and feces. Inflammatory infiltration damage was observed in the colonic epithelial tissue, and gut microbiota fluctuated with a decreased abundance of Lactobacilli in feces. Oral supplementation with Lactobacillus rhamnosus GG (LGG) lessened TiO2 NPs-induced colonic inflammatory injury, which might due to downregulation of nuclear factor kappa-B (NF-κB). Meanwhile, LGG maintained normal intestinal microbiome homeostasis, thereby improving TiO2 NPs-induced colon injury in juvenile rats. Moreover, fecal microbiota transplant (FMT) experiment indicated possible TiO2 NPs-induced intestinal microbiota disorder led to colonic inflammation. Our works suggested the urgent need for additional studies on the risk safety assessment, mechanism, and prevention of juvenile health damage from exposure to TiO2 NPs.


Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Lacticaseibacillus rhamnosus/crescimento & desenvolvimento , Nanopartículas/toxicidade , Probióticos/uso terapêutico , Titânio/toxicidade , Adulto , Animais , Criança , Fezes/química , Fezes/microbiologia , Feminino , Homeostase , Humanos , Inflamação , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Nanopartículas/metabolismo , Ratos , Titânio/metabolismo
10.
Toxicol Res (Camb) ; 9(4): 371-378, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32905215

RESUMO

Quantum dots (QDs) have recently attracted considerable attention in the biomedical fields because of their unique and excellent optical properties. However, information on their health effects, particularly in the reproductive system, is limited. The present study focuses on the effects of intravenous injection of CdSe/ZnS QDs on the reproductive system and embryo development at various stages of pregnancy in mice. The CdSe/ZnS QDs intravenously injected in mice during pregnancy accumulated in the maternal liver, uterus and placenta. This accumulation affected the growth and development of the embryo during the early and middle stages of pregnancy. Moreover, genotoxicity to the placenta after exposure to CdSe/ZnS QDs was demonstrated by the increased expression levels of genes related to oxidative stress and apoptosis and the reduced expression levels of genes related to the nutrient and waste transportation. Alterations in the gene expression levels have hindered the transport of metabolites across the placenta, which in turn affected the ability of the fetus to obtain nutrients.

11.
Food Chem Toxicol ; 143: 111552, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32640348

RESUMO

The growing use of zinc oxide nanoparticles (ZnO NPs) in various applications has raised many concerns about the potential risks to human health. In this research, the protective effects of cellular oxidative stress inhibitor N-Acetyl-cysteine (NAC) and endoplasmic reticulum (ER) stress inhibitor Salubrinal (Sal) on reproductive toxicity induced by ZnO NPs were investigated. The results showed that application of these two kinds of cell stress inhibitors after oral ingestion of ZnO NPs could prevent the weight loss of pregnant mice; reduce zinc content in the uterus, placenta and fetus; reduce abnormal development of the offspring; and decrease fetal abortion. Furthermore, RT-qPCR, Western blot and immunofluorescence assay results indicated that NAC restored the expression of Gclc, reduced the expression of ATF4, JNK and Caspase-12, and decreased the expression of eNOS and IGF-1, in the placenta. Sal decreased the expression of ATF4, JNK and Caspase-12, and increased the expression of eNOS and IGF-1caused by the oral ingestion of ZnO NPs. These results indicated that treatment with NAC and Sal after oral exposure could reduce reproductive and development toxicity caused by ZnO NPs which induced reproductive and development toxicity that was probably caused by the activation of oxide stress and ER stress.


Assuntos
Acetilcisteína/farmacologia , Cinamatos/farmacologia , Nanopartículas Metálicas/toxicidade , Tioureia/análogos & derivados , Óxido de Zinco/toxicidade , Animais , Feminino , Sequestradores de Radicais Livres/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Nanopartículas Metálicas/química , Camundongos , Gravidez , Distribuição Aleatória , Tioureia/farmacologia , Aumento de Peso , Óxido de Zinco/química
12.
J Microbiol ; 58(8): 668-674, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32583285

RESUMO

A multiplex polymerase chain reaction (mPCR) with propidium monoazide (PMA) and internal amplification control (IAC) for the simultaneous detection of waterborne pathogens Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, and Escherichia coli O157:H7, was developed. This PMA-IAC-mPCR assay used four new specific primers based on the genes for invA, ecfX, cesB, and fliC, respectively. A 16S rRNA primer was chosen for IAC to eliminate false negative results. The photosensitive dye, propidium monoazide (PMA) was used to exclude signals from dead bacteria that could lead to false positive results. In pure culture, the limits of detection (LOD) were 101 CFU/ml for P. aeruginosa, 102 CFU/ml for both Salmonella spp. and E. coli O157:H7, and 103 CFU/ml for B. cereus, respectively. In addition, with a 6-8 h enrichment of all four bacteria that were combined in a mixture that was spiked in water sample matrix, the LOD was 3 CFU/ml for Salmonella spp., 7 CFU/ml for E. coli O157:H7, 10 CFU/ml for B. cereus and 2 CFU/ml for P. aeruginosa. This PMA-IAC-mPCR assay holds potential for application in the multiplex assay of waterborne pathogens.


Assuntos
Bacillus cereus/isolamento & purificação , Carga Bacteriana/métodos , Escherichia coli O157/isolamento & purificação , Pseudomonas aeruginosa/isolamento & purificação , Salmonella/isolamento & purificação , Poluição da Água/análise , Azidas/farmacologia , Bacillus cereus/genética , Proteínas de Bactérias/genética , Escherichia coli O157/genética , Proteínas de Escherichia coli/genética , Flagelina/genética , Limite de Detecção , Reação em Cadeia da Polimerase Multiplex/métodos , Propídio/análogos & derivados , Propídio/farmacologia , Pseudomonas aeruginosa/genética , RNA Ribossômico 16S/genética , Salmonella/genética , Microbiologia da Água
13.
Environ Toxicol ; 35(9): 982-990, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32333507

RESUMO

In this work, we aimed to evaluate the adverse effects and the mechanism of intestinal barrier caused by titanium dioxide nanoparticles (TiO2 NPs). Here, the effects of two different dosages (300 and 1200 mg/kg) of TiO2 NPs on female mice (n = 5) were investigated. After 28-day oral exposure, the results of Ti content were significantly increased in the ileum in comparison with the control. The histopathological structure index of the ileum was significantly changed after TiO2 NPs exposure; villi height and crypt depth were decreased and increased, respectively. Meanwhile, TiO2 NPs treatment also significantly altered the transcription levels of genes. First, the GATA-3 and STAT-4 were upregulation and downregulation, respectively. Second, gene expressions of the Zonula Occludens-1, claudin (CLDN)-12, occludin, and myosin light chain kinase were significantly upregulated, while the CLDN-3 was decreased. Finally, the caspase-3, caspase-9, and caspase-12 were upregulated. The results of TUNEL staining indicated apoptosis in the ileum. In general, TiO2 NPs treatment significantly changed the intestine physical barrier in a dose-dependent manner. The toxicity of TiO2 NPs could be through the imbalance in the Th1/Th2.


Assuntos
Apoptose/efeitos dos fármacos , Íleo/efeitos dos fármacos , Nanopartículas/toxicidade , Equilíbrio Th1-Th2/efeitos dos fármacos , Titânio/toxicidade , Administração Oral , Animais , Relação Dose-Resposta a Droga , Feminino , Íleo/imunologia , Íleo/metabolismo , Íleo/patologia , Camundongos , Nanopartículas/ultraestrutura , Tamanho da Partícula , Propriedades de Superfície , Titânio/química , Titânio/farmacocinética
14.
Nanomaterials (Basel) ; 10(2)2020 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-32024284

RESUMO

ZnO NPs have been assessed to show adverse effects on reproductive organs, but the molecular mechanisms of reproductive toxicity have not been sufficiently studied. In this research, the dosage effects from the oral exposure of ZnO NPs (30 nm) to pregnant mice in gestation day 10.5 to 17.5 was analyzed. Pregnant mice exposed to ZnO NPs induced dam injury, mice fetal growth restriction, and the fetus number decreased. The pathological evaluation showed that ZnO NPs exposure caused placental spongiotrophoblast area decease and structural damage. The RT-qPCR and immunocytochemistry data indicated that ZnO NPs could induce placenta oxide stress, endoplasmic reticulum stress responses, apoptosis, and altered placental function. These findings indicated that ZnO NPs could induce dam injury and fetal growth restriction. Reproductive toxicity of ZnO NPs may be due to placental injury and function alteration caused by apoptosis, oxide stress, and endoplasmic reticulum stress after ZnO NPs exposure.

15.
J Dairy Sci ; 103(4): 3055-3065, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32037161

RESUMO

Cronobacter spp. are important opportunistic foodborne pathogens in powdered infant formula that cause many serious diseases in neonates and infants. In this study, a novel assay based on dual signal amplification strategy was developed by coupling asymmetric tailing PCR (AT-PCR) with rolling circle amplification (RCA) for the detection of Cronobacter spp. in milk. The tailing single-stranded DNA was generated through AT-PCR and used to initiate RCA, generating tandem repetitive G-quadruplex sequences. In the presence of the fluorescence dye thioflavin T that could intercalate into the G-quadruplex structures, the fluorescence signal was detected with a microplate reader. The AT-PCR coupled with RCA assay was specific for Cronobacter spp. detection because of the highly specific primers chosen for the AT-PCR. The limits of detection were 4.3 × 101 cfu/mL in pure culture and 4.5 × 102 cfu/mL in spiked milk, respectively. The fixed sequences designed in the hairpin DNA allowed this AT-PCR coupled with RCA assay to serve as a universal platform for the detection of other pathogens by modifying the specificity of the PCR primers.


Assuntos
Benzotiazóis/análise , Cronobacter/isolamento & purificação , Leite/microbiologia , Reação em Cadeia da Polimerase/veterinária , Animais , Cronobacter/genética , DNA , Primers do DNA/genética , Fluorescência , Quadruplex G , Sensibilidade e Especificidade
16.
Int J Nanomedicine ; 14: 9563-9576, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31824151

RESUMO

PURPOSE: The aim of this study was to evaluate the adverse effects of ZnO NPs on male reproductive system and explore the possible mechanism. METHODS: In this study, the effect of oral administration of 50, 150 and 450 mg/kg zinc oxide nanoparticles (ZnO NPs) in adult male mice was studied over a 14-day period. RESULTS: The results showed that the number of sperms in the epididymis and the concentration of testosterone in serum were decreased with an increased dose of ZnO NPs. Testicular histopathological lesions like detachment, atrophy and vacuolization of germ cells were observed. The results showed that increased dosage of ZnO NPs correspondingly up-regulated the IRE1α, XBP1s, BIP, and CHOP (P<0.05) which are genes related to ER stress. These observations indicated that ZnO NPs had adverse effects on the male reproductive system in a dose-dependent manner possibly through ER stress. The expression of caspase-3 was significantly increased in all the treated groups (P<0.001), which reflected the possible activation of apoptosis. Additionally, there was significant down-regulation of the gene StAR (P<0.05), a key player in testosterone synthesis. When an ER-stress inhibitor salubrinal was administered to the 450 mg/kg ZnO NPs treatment group, the damages to the seminiferous tube and vacuolization of Sertoli and Leydig cells were not observed. Furthermore, the testosterone levels in the serum were similar to the control group after the subsequent salubrinal treatment. CONCLUSION: It may be inferred that the ZnO NP's reproductive toxicity in male mice occurred via apoptosis and ER-stress signaling pathway.


Assuntos
Estresse do Retículo Endoplasmático/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Testículo/efeitos dos fármacos , Óxido de Zinco/toxicidade , Animais , Apoptose/efeitos dos fármacos , Relação Dose-Resposta a Droga , Estresse do Retículo Endoplasmático/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células Intersticiais do Testículo/efeitos dos fármacos , Masculino , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/efeitos adversos , Camundongos , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/patologia , Transdução de Sinais/efeitos dos fármacos , Contagem de Espermatozoides , Testículo/metabolismo , Testículo/patologia , Testosterona/sangue , Zinco/farmacocinética , Óxido de Zinco/administração & dosagem , Óxido de Zinco/efeitos adversos
17.
IET Nanobiotechnol ; 13(1): 6-11, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30964030

RESUMO

Circulating tumour cells (CTCs) draw significant attention as a promising biomarker for cancer prognosis, status monitoring, and metastasis diagnosis. However, the concentration of CTCs in peripheral blood is usually extremely low, thereby requiring enrichment followed by isolation of CTCs prior to detection. An immunomagnetic separation is a promising tool for CTCs enrichment. In this study, a cost-effective magnetic separation method, based on streptavidin-biotin complexation, was developed and the effects of magnetic beads' size in CTCs capture were compared. Magnetic nanobeads which were 25 nm in diameter lead to highest capture efficiency (82.2%) compared with 150 nm magnetic beads and 1 µm microbeads. Based on the streptavidin-biotin system, 25 nm magnetic nanobeads could capture model CTCs over 80% efficiency even at concentrations as low as ∼25 cells/mL that may represent the actual level of CTCs in peripheral blood of cancer patients. Furthermore, the isolated cells remained robust and healthy showing insignificant changes in morphology and behaviour when cultured for 24 h immediately after capture and isolation. The magnetic nanobeads based on streptavidin-biotin complexation showed promise for the easy and efficient capture and isolation of healthy CTCs for further diagnosis and analysis.


Assuntos
Proteínas de Bactérias/química , Biotina/análogos & derivados , Separação Imunomagnética/métodos , Nanopartículas de Magnetita/química , Células Neoplásicas Circulantes , Proteínas de Bactérias/metabolismo , Biotina/química , Biotina/metabolismo , Citometria de Fluxo , Humanos , Células K562 , Células MCF-7 , Tamanho da Partícula
18.
J Dairy Sci ; 102(6): 4945-4953, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30904294

RESUMO

A fluorescence assay combined with PCR, catalytic hairpin assembly (CHA), and graphene oxide (GO) was established to detect emetic Bacillus cereus in milk samples. The processes of the assay are not new, but components of the processes make the assay useful. Two partially complementary hairpin probes (H1 and FAM-H2) were designed according to the target single-strand DNA (ssDNA). The CHA reaction could be initiated only by the target ssDNA, which was generated by the denaturation of PCR amplicons. In the absence of the target ssDNA, CHA reaction could not be triggered, which caused the H1 and FAM-H2 adsorbing on the surface of GO and exhibiting a low fluorescence intensity. Addition of the target ssDNA resulted in opening of the hairpin H1 that subsequently hybridized with H2. Then, target ssDNA would be replaced from the H1 and recycled to promote another CHA reaction. Through the CHA reaction, multiple H1-H2 duplexes were generated that could not adsorb on the surface of GO. Thus, a strong fluorescence signal would be obtained. The assay showed a limit of detection for emetic B. cereus of 6.2 × 101 cfu/mL in pure culture and 5.9 × 102 cfu/mL in spiked milk without enrichment. By changing the PCR primer, the assay developed in this study had potential to detect other bacteria.


Assuntos
Bacillus cereus/isolamento & purificação , Grafite , Leite/microbiologia , Animais , Bacillus cereus/genética , Bacillus cereus/patogenicidade , Técnicas Biossensoriais , Catálise , DNA Bacteriano/química , DNA Bacteriano/isolamento & purificação , Fluorescência , Sequências Repetidas Invertidas , Reação em Cadeia da Polimerase
19.
J Pharm Biomed Anal ; 169: 181-187, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-30877929

RESUMO

A new, facile, low-cost, and highly sensitive method for detection of Listeria monocytogenes involving a combination of asymmetric polymerase chain reaction (aPCR) and rolling circle amplification (RCA) had been developed. The aPCR-RCA processes were not new but components of the processes made the assay useful. Twenty-one thymine (21-T) tagged forward primer generated universal twenty-one adenine (21-A) aPCR amplicons after aPCR amplification. A poly-T sequence dumbbell-like RCA template was produced through the blunt-end ligation activity of T4 DNA ligase. After the mixture of aPCR amplicons and dumbbell-like RCA template, the RCA reaction would initiate when the addition of phi29 DNA polymerase, then a large number of G-quadruplex sequences were produced which allowed the intercalation of Thioflavin T (3,6-dimethyl-2-(4-dimethylaminophenyl) benzo-thiazolium cation, THT) for easy fluorescence detection. Under the optimal conditions, the assay showed a limit of detection (LOD) of 4.8 × 101 CFU/mL in pure culture and 4.0 × 102 CFU/g in spiked lettuce homogenates. By changing the aPCR primer, the aPCR-RCA method developed in this study had a potential to detect other bacteria without the design an RCA template for each bacterium.


Assuntos
Bioensaio/métodos , Técnicas Biossensoriais/métodos , Corantes Fluorescentes/química , Listeria monocytogenes/química , Reação em Cadeia da Polimerase/métodos , Benzotiazóis/química , Primers do DNA/química , Primers do DNA/genética , Fluorescência , Quadruplex G , Limite de Detecção , Listeria monocytogenes/genética
20.
Nanomaterials (Basel) ; 9(2)2019 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-30769773

RESUMO

The toxicity, especially the transgenerational toxicity of quantum dots (QDs) in vivo, is still scarcely understood in spite of great promising applications of QDs in biomedicine. In this study, the maternal status, pregnancy outcome, and fetus development of parental generation (P0) to offspring in three generations (F3) were investigated after Kunming mice perinatal (GD 13-PND 5) exposure to Cd containing QDs (CdSe/ZnS QDs) and CdCl2. The results show CdSe/ZnS QDs induced placenta injuries in P0 and diminished placenta diameters in F1 and F2. Bodyweight growth decreased in the CdSe/ZnS QDs treatment group in the F1 and F2 generation. Additionally, CdSe/ZnS QDs significantly altered the expression of key genes in the Shh signal pathway. Overall, this study exhibited that the CdSe/ZnS QDs exposure during perinatal period impaired placenta growth in the first two generations, but not on the third generation. The toxicological actions of the CdSe/ZnS QDs might be through the effects on the Shh signal pathway.

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