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@#Abstract: Objective In order to provide reference for emergency treatment of a sudden food poisoning incident, pathogen detection and drug resistance analysis were carried out. Methods Diarrheal stool and surplus food samples were detected by GB 4789 and the isolates were identified by VITEK2 and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), at the same time, the bacterial drug sensitivity test was carried out by using the method of microbroth dilution, and the isolates from different sources were molecularly classified by pulsed field gel electrophoresis (PFGE), and the correlation between the strains was analyzed by BioNumerics software. Results Totaly 13 leftovers and 3 diarrhea patients were isolated and identified, The total number of colonies and coliforms in 7 leftovers samples all exceeded the standard, and Citrobacter freundii was detected in 5 leftovers and 2 stools. The results of drug sensitivity test showed that seven strains of Citrobacter freundii were sensitive to ciprofloxacin, tetracycline, chloramphenicol, gentamicin, amikacin, cefotaxime and meropenem, but completely resistant to ampicillin, and there was no multiple drug resistance. The results of pulsed field gel electrophoresis (PFGE) showed that 7 strains of Citrobacter freundii had the same PFGE bands and 100% homology, showing the same clone. Conclusions This food poisoning incident was caused by Citrobacter freundii. The pathogen of food poisoning can be quickly and accurately determined by MALDI-TOF MS, which is beneficial to the early diagnosis and treatment of infectious diseases. It is suggested to strengthen the corresponding management, improve food safety awareness and prevent similar incidents.
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Background The incidence of Legionnaires' disease is increasing globally and artificial water environment is becoming a common source of outbreaks. Molecular typing techniques can help prevent and control Legionella. Objective To understand the molecular epidemiological characteristics of Legionella pneumophila in artificial water environment of Shanghai hospitals, and provide a scientific basis for the prevention and control of Legionnaires' disease. Methods Water samples were collected from artificial water environment in 14 hospitals from May to October each year from 2019 to 2020 in Shanghai. A total of 984 water samples were collected from 8 Grade-A tertiary hospitals and 6 non-Grade-A tertiary hospitals, including 312 samples of cooling water, 72 samples of chilled water, and 600 samples of tap water. The water samples were isolated and serotyped for Legionella pneumophila and preserved, and the positive rate of Legionella pneumophila in the samples was used as an indicator of contamination. The preserved strains were resuscitated and 81 surviving strains were obtained for pulsed field gel electrophoresis (PFGE) typing analysis. Results A total of 124 Legionella pneumophila positive water samples were detected, with a positive rate of 12.60%. The positive rate was higher in the Grade-A tertiary hospitals (16.54%, 87/526) than in the non-Grade-A tertiary hospitals (8.08%, 37/458) (χ2=15.91, P<0.001). The positive rate of cooling water (23.40%) was the highest among different types of water samples, and the difference was statistically significant (χ2=61.19, P<0.001). The difference in positive rate of tap water was statistically significant among different hospital departments (χ2=11.37, P<0.05). The positive rate in 2019 (15.06%) was higher than that in 2020 (9.84%) (χ2=6.23, P<0.05). From May to October, August had the highest annual average positive rate (16.46%) and October had the lowest (8.54%), but the difference in positive rates among months was not statistically significant (χ2=5.39, P=0.37). The difference in positive rate among districts was statistically significant (χ2=24.88, P<0.001). A total of 131 strains of Legionella pneumophila were isolated, with serotype 1 (80.15%, 105/131) predominating. Among the 81 surviving strains of Legionella pneumophila subjected to PFGE typing, the band-based similarity coefficients ranged from 41.30% to 100%. Among the 29 PFGE band types (S1-S29) recorded, each band type included 1-10 strains, and S28 was the dominant band type. Four clusters (I-IV) of PFGE band types were identified, accounting for 66.67% (54/81) of all strains and containing 13 band types. Conclusion Legionella pneumophila contamination is present in the artificial water environment of hospitals in Shanghai from 2019 to 2020, and the contamination in tap water deserves attention. The detected serotype of Legionella pneumophila is predominantly type 1, and PFGE typing reveals the presence of genetic polymorphism. Therefore, the monitoring and control of Legionella pneumophila in hospital artificial water environment should be strengthened.
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Objective To investigate the etiological characteristics of food poisoning isolates of Vibrio parahaemolyticus (VP) from 2019 to 2021 in Zhongshan City. Methods A total of 37 strains of Vibrio parahaemolyticus isolated from 8 food poisoning incidents in Zhongshan City from 2019 to 2021 were collected, including 1 residual food isolate and 36 human isolates. The genetic correlation of Vibrio parahaemolyticus food poisoning isolates in this region was analyzed by serological typing, virulence gene detection (TLH, TDH, and TRH), drug sensitivity test, pulsed field gel electrophoresis (PFGE) and multipoint sequence typing (MLST). Results The 37 strains of Vibrio parahaemolyticus were divided into 4 serotypes: O3:K6, O10:K4, O4:K8, and O4:KUT. The tdh+ and trh- were the main virulence genotypes, accounting for 97.30% (36/37). The drug resistance rate of cefazolin was 40.54% (15 strains R, 22 strains I), and no multidrug-resistant strains were found. The 37 VP strains were divided into 23 PFGE types and 6 cluster groups, with correlation coefficients ranging from 60.4%-100%. The multipoint sequencing typing showed that the 37 VP strains were divided into 9 ST types and 3 complex groups, of which ST3 type was the main type (23 strains, 62.1%). Conclusion This study has found that the dominant virulence types of Vibrio parahaemolyticus food poisoning isolates in Zhongshan City from 2019 to 2021 are tdh+ and trh-, and 37 representative strains can be divided into 6 PFGE clusters and 9 ST types with MLST type being mainly ST3. This study has identified the rare serotype O10:K4 which has caused an increase in the proportion of food poisoning events, suggesting that we should strengthen detection and be alert to the risk of continued local epidemics of new rare serotype strains.
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@#Abstract: Objective To analyze the drug sensitivity and the carrying of carbapenem resistant gene of Acinetobacter baumannii isolated from clinical patients and clinical objects, and analyze the homology of strains to provide support for the control of nosocomial infection. Methods A total of 38 strains of Acinetobacter baumannii isolated from patients and clinical objects surface were collected from January 2019 to August 2020. The antimicrobial susceptibility was tested by the minimum inhibitory concentration method. In addition, the resistance related genes were detected by polymerase chain reaction method, and homology analysis was performed by enterobacterial repetitive Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR). Results All 34 strains of Acinetobacter baumannii isolated from Clinical patients and 4 strains isolated from clinical objects carried blaOXA-51 and imp resistance genes, neither of them carried blaVIM gene. 32 Acinetobacter baumannii carrying blaOXA-23 gene, 28 strains carrying blaTEM gene, 7 strains carrying blaOXA-58 gene. After cluster analysis, 38 Acinetobacter baumannii isolates were classified into 7 genotypes (expressed A, B, C, D, E, F, G), and cluster E and cluster G were the main clusters, containing 12 strains (12/38, 31.6%) and 18 strains (18/38, 47.4%), respectively, as the main prevalent clonal strains. Conclusions Acinetobacter baumannii isolated from different sources have the significant differences in drug resistance and carry different resistance genes. There is no direct correlation between patients and environmental isolates of Acinetobacter baumannii belonging to different clonal strains. Also, there aren’t significant correlation between clinical patients infected with Acinetobacter baumannii.
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ObjectiveThe therapeutic effect of polysaccharides from Zanthoxyli Pericarpium on Alzheimer's disease（AD） was evaluated through establishing a mouse model of AD， and the structural characteristics of the polysaccharides was analyzed by sugar spectrum. MethodThe AD model of mice with rapid aging was established by intraperitoneal injection of D-galactose combined with gavage of aluminum trichloride， and the learning and memory ability of mice was evaluated by Morris water maze test， the histopathological status of brain and neuronal damage were observed by hematoxylin-eosin（HE） staining and Nissl staining. After hydrolysis of polysaccharides from Zanthoxyli Pericarpium with acid and different glycosidases， the characteristics of hydrolysates were analyzed by high performance thin layer chromatography（HPTLC） and fluorescence assisted carbohydrate gel electrophoresis（PACE）. HPTLC chromatography was performed on a silica gel 60 plate with sampling volume of 5 μL， developing solvent of ethyl acetate-glacial acetic acid-water（2∶2∶1）， developing twice， aniline-diphenylamine-phosphoric acid solution as chromogenic agent， and heating at 105 ℃ for 10 min， and then observed under sunlight. PACE experimental conditions were 34% separation gel and 8% concentration gel， electrophoresis buffer was 0.1 mol·L-1 tris（hydroxymethyl） aminomethane（Tris）-boric acid buffer（pH 8.2）. Electrophoresis was carried out at 0 ℃ and the loading amount was 3-6 μL. The sample ran to the front of the gel with a constant current of 15 mA， and imaged under ultraviolet 365 nm. ResultThe results of Morris water maze test showed that polysaccharides from Zanthoxyli Pericarpium significantly improved the learning and memory ability of AD model mice， shortened the escape latency， and significantly increased the number of crossing and the residence time in the target quadrant. The results of histopathological experiments showed that polysaccharides from Zanthoxyli Pericarpium could improve the pathological conditions and neuronal damage in the CA1 and CA3 regions of hippocampus of AD mice， and the number of Nissl corpuscles was significantly increased. The results of sugar spectrum analysis showed that the results of HPTLC and PACE analysis were basically consistent， polysaccharides from Zanthoxyli Pericarpium could be mainly hydrolyzed into small molecular sugars by cellulase and pectinase， indicating that they mainly contained β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， and could be slightly hydrolyzed by glucanase， β-galactosidase and β-mannase， indicating that they contained only a small amount of α-1，6-glucosidic bond， β-galactosidic bond， β-1，4-mannosidic bond. ConclusionPolysaccharides from Zanthoxyli Pericarpium has obvious therapeutic effect on AD mice， and its structure mainly contains β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， which can provide a reference for the structural analysis of traditional Chinese medicine polysaccharides.
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In criminal investigations, postmortem interval (PMI) is important information to be inferred in homicide investigations, as well as the focus and the difficulty in forensic pathology research. Because the DNA content in different tissues is relatively constant and shows changes regularly with the extension of PMI, it has become a research hotspot of PMI estimation. This paper reviews the recent progress of PMI estimation technologies including DNA-based single cell gel electrophoresis, image analysis, flow cytometry, real-time fluorescence quantitative PCR and high-throughput sequencing, hoping to provide references for forensic medicine practice and scientific research.
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Humans , Postmortem Changes , Autopsy/methods , DNA/genetics , Forensic Medicine , Forensic PathologyABSTRACT
Introduction@#PCR to detect and amplificate the virulence genes of STEC is specific and more sensitive, however, it takes five to six hours for whole test procedure and requires special lab instruments such as thermocycler. Loop-mediated isothermal amplification is a simple, rapid, specific and cost-effective nucleic acid amplification method by using four to six primers when compared to PCR, nucleic acid sequence-based amplification, self-sustained sequence replication and strand displacement amplification.@*Goal@#Detection and comparison of STEC by PCR and LAMP@*Materials and Methods@#In our study, we analyzed comparison of PCR and LAMP results on standard strain used quality control strain solution which diluted 1pg/µL DNA, 10 pg/µL DNA, 100 pg/µL DNA, 1 ng/μL DNA, 10 ng/μL DNA, and 100 ng/μL DNA concentration from LB agar cultures. @*Research ethics@#Permission to submit the survey was granted by the Ethics Review Committee of the MNUMS and the survey was conducted in accordance with the rules and regulations.@*Result@#Sensitivity of Stx1 and stx2 genes in PCR results are positive in 10 pg/µL DNA solution and negative in 1pg/µL DNA. In LAMP test results showed that positive for all concentration. It shows that LAMP method sensitivity is 10 times more than PCR. @*Conclusion@#It shows that LAMP method sensitivity is 10 times more than PCR. All in allLAMP test is cost effective test with sensitive for detection STEC.
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Introduction@#PCR to detect and amplificate the virulence genes of STEC is specific and more sensitive, however, it takes five to six hours for whole test procedure and requires special lab instruments such as thermocycler. Loop-mediated isothermal amplification is a simple, rapid, specific and cost-effective nucleic acid amplification method by using four to six primers when compared to PCR, nucleic acid sequence-based amplification, self-sustained sequence replication and strand displacement amplification.@*Materials and Methods@#In our study, we analyzed comparison of PCR and LAMP results on standard strain used quality control strain solution which diluted 1pg/µL DNA, 10 pg/µL DNA, 100 pg/µL DNA, 1 ng/μL DNA, 10 ng/μL DNA, and 100 ng/μL DNA concentration from LB agar cultures. @*Research ethics@#Permission to submit the survey was granted by the Ethics Review Committee of the MNUMS and the survey was conducted in accordance with the rules and regulations. @*Goal@#Detection and comparison of STEC by PCR and LAMP@*Result@#Sensitivity of Stx1 and stx2 genes in PCR results are positive in 10 pg/µL DNA solution and negative in 1pg/µL DNA. In LAMP test results showed that positive for all concentration. It shows that LAMP method sensitivity is 10 times more than PCR. @*Conclusion@#It shows that LAMP method sensitivity is 10 times more than PCR. All in allLAMP test is cost effective test with sensitive for detection STEC.
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Objective@#To analyze the etiological characteristics of an outbreak of Campylobacter foodborne disease in a middle school in Suzhou City, so as to provide insights into the identification of pathogenic factors of Campylobacter foodborne disease outbreaks.@*Methods@#Eighteen anal swabs from patients, 10 anal swabs from canteen workers, 43 food samples, 2 drinking water samples, 2 food original material samples and 31 environmental samples were collected, and the pathogens were rapidly screened using the gastrointestinal infection detection strip. The pathogens were isolated and cultured using the double-pore filtration membrane method, and cluster analysis of bacterial isolates was performed using pulsed field gel electrophoresis ( PFGE ). In addition, the susceptibility of Campylobacter isolates to antibiotics was tested using the Campylobacter agar dilution method.@*Results@#A total of 63 cases with Campylobacter infections were reported, and the major clinical symptoms included diarrhea ( 51 cases, 80.95% ) and fever ( 39 cases, 61.90% ), while no inpatients or deaths were found. Twelve Campylobacter-positive samples were detected, including 11 anal swabs sampled from patients and one food original material sample. Among the 11 positive anal swabs, there were 10 samples positive for Campylobacter jejuni and one sample positive for C. coli, and of the one positive food original material, C. coli was identified. PFGE analysis showed that 10 C. jejuni isolates of had 100.0% homology, and these 10 isolates were 100.0% resistant to naphthyridic acid, ciprofloxacin and tetracycline, appearing multidrug resistance.@*Conclusions@#This is an outbreak of foodborne disease caused by C. jejuni infections. Gastrointestinal infection detection strips, double-pore filtration membrane and PFGE typing are rapid and accurate to identify pathogenic factors.
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ObjectiveTo determine the pathogenic cause in a foodborne diseases outbreak of Salmonella enteritidis in a company in Suzhou City， and provide evidence for epidemiological investigation and guidance for clinical treatment. MethodsRelevant specimens were examined for Salmonella， Shigella， Staphylococcus aureus and Vibrio parahaemolyticus. Furthermore， for the isolated Salmonella enteritidis， a micro broth dilution method was used for antimicrobial susceptibility testing， and pulsed field gel electrophoresis （PFGE） was used for molecular typing. ResultsA total of 44 strains of Salmonella enteritidis were detected from 43 anal swabs of the patients in the outbreak， 7 anal swabs of canteen employees， 31 retained food specimens and 6 environmental specimens. A total of 15 antimicrobial susceptibility testings showed that the 44 strains had the same antimicrobial resistance spectrum， which was 100% resistant to cefazolin， ampicillin， ampicillin/sulbactam， polymyxin E and nalidixic acid， suggesting a multi-drug resistance to more than three antibiotics. PFGE cluster analysis showed that the 44 strains had a 100% of genetic similarity. ConclusionThe outbreak is caused by the consumption of food contaminated with Salmonella enteritidis. The isolated strains have multi-drug resistance， which could guide appropriate antimicrobial treatment based on the antimicrobial susceptibility testing.
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Background: Saliva being an important biological fluid of our body contains both specific and nonspecific protective factors which form an integral part of our immune system. Salivary proteins play a substantial role in protecting humans against infection. Their level in oral cavity is subject to constant variations which is dependent on various factors. Purpose: The purpose of the study was to compare the levels of salivary proline-rich proteins (PRPs), glycoproteins, amylase bands, and salivary pH in children with early childhood caries before and after treatment using gel electrophoresis. Materials and Methods: The whole salivary pH, mean protein concentrations, and electrophoretic profiles of the salivary proteins were measured using sodium dodecylsulfate–polyacrylamide gel electrophoresis in both pre- and posttreatment groups. Statistical analysis was done using Statistical Package for Social Sciences (SPSS) version 15.0 software. Chi-square test and independent t-test were used to further compare the results. Results: The results were statistically significant in all the groups. There was variation in pre- and posttreatment values seen. Conclusion: Salivary proteins (glycoproteins, PRPs, and amylase) establish an imperative defense mechanism as antimicrobial agents.
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Some antioxidant compounds have a pro-oxidant effect in the presence of transition metal ions, due to the reduction of Mn+ to M(n-1)+ with simultaneous formation of free radicals, which then promote DNA damage. In the present study, we evaluated the pUC19 DNA damage in a solution containing Cu(II) and ascorbic acid (AA) or S(IV) saturated with air by agarose gel electrophoresis. Our results showed that this damage decreases if AA and S(IV) are simultaneously added. This study also illustrates the importance of Cu(II) in this process, as no DNA damage was observed when AA or S(IV) were present in the absence of this metallic ion. Our data showed that DNA preservation depends on the concentration of AA and S(IV) and occurs when the [S(IV)]:[AA] ratio ranges from 1:1 to 20:1. Absorbance measurements and thermodynamic data show that no reaction occurs between AA and S(IV) when this mixture (pH 5.5) is added to pUC-19 DNA. The presence of dissolved oxygen may be the cause of AA consumption in the mixture of these two antioxidants, which subsequently decreases DNA damage.
Subject(s)
Ascorbic Acid/adverse effects , Sulfites , DNA Damage , Copper/pharmacology , Ions/adverse effects , Antioxidants/adverse effects , Electrophoresis, Agar Gel/instrumentation , Free Radicals/pharmacology , Hydrogen-Ion ConcentrationABSTRACT
We developed a high-efficiency microfluidic chip for extracting exosomes from human plasma. We collected peripheral blood from normal human, designed and fabricated a microfluidic chip based on nanoporous membrane and agarose gel electrophoresis to isolate exosomes. The extracted exosomes were characterized by transmission electron microscopy, nanosight and Western blotting, the morphology, concentration and particle size of exosomes were identified and analyzed. Meanwhile, we used ultracentrifugation and microfluidic chip to isolate exosomes separately. The particle size and concentration of the exosomes extracted by two methods were compared and analyzed, and their respective extraction efficiency was discussed. Finally, the expression level of miRNA-21 in exosomes was analyzed by RT-PCR. The microfluidic chip isolated (in 1 hour) high-purity exosomes with size ranging from 30-200 nm directly from human plasma, allowing downstream exosomal miRNA analysis. By comparing with ultracentrifugation, the isolation yield of microfluidic chip was 3.80 times higher than ultracentrifugation when the volume of plasma sample less than 100 μL. The optimized parameters for exosome isolation by gel electrophoresis microfluidic chip were: voltage: 100 V; concentration of agarose gel: 1.0%; flow rate of injection pump: 0.1 mL/h. The gel electrophoresis microfluidic chips could rapidly and efficiently isolate the exosomes, showing great potential in the research of exosomes and cancer biomarkers.
Subject(s)
Humans , Exosomes , MicroRNAs/genetics , Microfluidics , Plasma , UltracentrifugationABSTRACT
ABSTRACT: Excessive infection and inflammation are the most common complications associated with castration. The objective of this study was to compare the efficacy of flunixin meglumine (FM), meloxicam (MX), or firocoxib (FX) for inflammation control after castration in horses using acute-phase proteins (APP) as markers of inflammation. Thirty healthy, unbroken, mixed-breed horses (body weight 358.62±45.57kg and age 4.99±2.63 years) were randomly (n=10 animals/group) allocated to receive one of three different post-castration anti-inflammatory medicines: Group 1 (FM 1.1mg/kg bwt, IV, s.i.d for 5 days); Group 2 (MX 0.6mg/kg bwt, IV, s.i.d for 5 days); and Group 3 (FX 0.1mg/kg bwt, IV, s.i.d for 5 days). All horses were castrated in standing position, using the open technique. Serum and peritoneal APP concentrations were measured by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and determined before castration (0), and 3, 5, 24, 48, 72, 120 and 168 hours after castration. The results were submitted to analysis of variance using the SAS statistical program, and means were compared by the Student-Newman-Keuls test (p 0.05). Three animals from the MX group developed hyperthermia (with rectal temperatures of 39.8, 39.3 and 38.9°C on day 4, 5 and 6, respectively) and showed local clinical signs of inflammation (inguinal and excessive scrotal edema) and reluctance to walk, as well as a rigid gait of the hind limbs. The same complications were observed in one FX horse. No complications were observed among the FM animals. The castration resulted in significant changes in serum and peritoneal values of total proteins, ceruloplasmin (Cp), transferrin (Tf), albumin (Alb), haptoglobin (Hp) and 1-acid glycoprotein (Gp) in animals of all experimental groups. However, the animals of the MX and FX groups presented more intense acute phase response compared to the animals of the FM group. Changes in the APP were associated with the surgical trauma of castration, but the differences between groups were associated with the ability of the nonsteroidal anti-inflammatory drug to control the inflammation. In conclusion, and based on the findings of acute phase proteins, flunixin is more efficient to control the magnitude of inflammation following castration as compared to meloxicam and firocoxib.
RESUMO: Infecção e inflamação excessivas são as complicações mais comuns associadas à castração. O objetivo deste estudo foi comparar a eficácia do flunixin meglumine (FM), meloxicam (MX) ou firocoxib (FX) no controle da inflamação após a castração em cavalos usando proteínas da fase aguda (APP) como marcadores de inflamação. Trinta equinos saudáveis (358,62±45,57kg; 4,99±2,63 anos) foram em função dos anti-inflamatórios utilizados após as castrações aleatoriamente (n= 10 animais/grupo) alocados em três diferentes grupos: Grupo 1 (FM 1,1mg/kg de peso, IV, sid por 5 dias); Grupo 2 (MX 0,6mg/kg de peso, IV, s.i.d por 5 dias); e Grupo 3 (FX 0,1mg/kg de peso, IV, s.i.d por 5 dias). Todos os cavalos foram castrados em posição quadrupedal, utilizando a técnica aberta. As concentrações de APP sérica e peritoneal foram separadas por eletroforese em gel de poliacrilamida (PAGE) com dodecil-sulfato de sódio (SDS) e determinadas no momento 0 (antes da castração) e com 3, 5, 24, 48, 72, 120 e 168 horas após a castração. Os resultados foram submetidos à análise de variância pelo programa estatístico SAS e as médias foram comparadas pelo teste de Student-Newman-Keuls (p 0,05). Três animais do grupo MX desenvolveram hipertermia (com temperatura retal de 39,8, 39,3 e 38,9° C nos dias 4, 5 e 6, respectivamente) e mostraram sinais clínicos locais de inflamação (edema inguinal e escrotal excessivo) e relutância em andar, bem como marcha rígida dos membros posteriores. As mesmas complicações foram observadas em um cavalo do FX. Não foram observadas complicações entre os animais do FM. Independente do grupo, a castração resultou em alterações significativas nos valores séricos e peritoneais de proteínas totais, ceruloplasmina (Cp), transferrina (Tf), albumina (Alb), haptoglobina (Hp) e glicoproteína ácida 1 (Gp). No entanto, os animais dos grupos MX e FX apresentaram resposta de fase aguda mais intensa quando comparados aos animais do FM. Alterações na resposta de fase aguda deveram-se ao trauma cirúrgico da castração, mas as diferenças entre os grupos foram associadas à capacidade do anti-inflamatório em controlar a inflamação. Em conclusão, baseado da resposta de fase aguda, o flunixin em comparação com o meloxicam e o firocoxib é mais eficiente no controle da inflamação após a castração em equinos.
ABSTRACT
Excessive infection and inflammation are the most common complications associated with castration. The objective of this study was to compare the efficacy of flunixin meglumine (FM), meloxicam (MX), or firocoxib (FX) for inflammation control after castration in horses using acute-phase proteins (APP) as markers of inflammation. Thirty healthy, unbroken, mixed-breed horses (body weight 358.62±45.57kg and age 4.99±2.63 years) were randomly (n=10 animals/group) allocated to receive one of three different post-castration anti-inflammatory medicines: Group 1 (FM 1.1mg/kg bwt, IV, s.i.d for 5 days); Group 2 (MX 0.6mg/kg bwt, IV, s.i.d for 5 days); and Group 3 (FX 0.1mg/kg bwt, IV, s.i.d for 5 days). All horses were castrated in standing position, using the open technique. Serum and peritoneal APP concentrations were measured by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and determined before castration (0), and 3, 5, 24, 48, 72, 120 and 168 hours after castration. The results were submitted to analysis of variance using the SAS statistical program, and means were compared by the Student-Newman-Keuls test (p<0.05). Three animals from the MX group developed hyperthermia (with rectal temperatures of 39.8, 39.3 and 38.9°C on day 4, 5 and 6, respectively) and showed local clinical signs of inflammation (inguinal and excessive scrotal edema) and reluctance to walk, as well as a rigid gait of the hind limbs. The same complications were observed in one FX horse. No complications were observed among the FM animals. The castration resulted in significant changes in serum and peritoneal values of total proteins, ceruloplasmin (Cp), transferrin (Tf), albumin (Alb), haptoglobin (Hp) and α1-acid glycoprotein (Gp) in animals of all experimental groups. However, the animals of the MX and FX groups presented more intense acute phase response compared to the animals of the FM group. Changes in the APP were associated with the surgical trauma of castration, but the differences between groups were associated with the ability of the nonsteroidal anti-inflammatory drug to control the inflammation. In conclusion, and based on the findings of acute phase proteins, flunixin is more efficient to control the magnitude of inflammation following castration as compared to meloxicam and firocoxib.(AU)
Infecção e inflamação excessivas são as complicações mais comuns associadas à castração. O objetivo deste estudo foi comparar a eficácia do flunixin meglumine (FM), meloxicam (MX) ou firocoxib (FX) no controle da inflamação após a castração em cavalos usando proteínas da fase aguda (APP) como marcadores de inflamação. Trinta equinos saudáveis (358,62±45,57kg; 4,99±2,63 anos) foram em função dos anti-inflamatórios utilizados após as castrações aleatoriamente (n= 10 animais/grupo) alocados em três diferentes grupos: Grupo 1 (FM 1,1mg/kg de peso, IV, sid por 5 dias); Grupo 2 (MX 0,6mg/kg de peso, IV, s.i.d por 5 dias); e Grupo 3 (FX 0,1mg/kg de peso, IV, s.i.d por 5 dias). Todos os cavalos foram castrados em posição quadrupedal, utilizando a técnica aberta. As concentrações de APP sérica e peritoneal foram separadas por eletroforese em gel de poliacrilamida (PAGE) com dodecil-sulfato de sódio (SDS) e determinadas no momento 0 (antes da castração) e com 3, 5, 24, 48, 72, 120 e 168 horas após a castração. Os resultados foram submetidos à análise de variância pelo programa estatístico SAS e as médias foram comparadas pelo teste de Student-Newman-Keuls (p<0,05). Três animais do grupo MX desenvolveram hipertermia (com temperatura retal de 39,8, 39,3 e 38,9° C nos dias 4, 5 e 6, respectivamente) e mostraram sinais clínicos locais de inflamação (edema inguinal e escrotal excessivo) e relutância em andar, bem como marcha rígida dos membros posteriores. As mesmas complicações foram observadas em um cavalo do FX. Não foram observadas complicações entre os animais do FM. Independente do grupo, a castração resultou em alterações significativas nos valores séricos e peritoneais de proteínas totais, ceruloplasmina (Cp), transferrina (Tf), albumina (Alb), haptoglobina (Hp) e glicoproteína ácida α1 (Gp). No entanto, os animais dos grupos MX e FX apresentaram resposta de fase aguda mais intensa quando comparados aos animais do FM. Alterações na resposta de fase aguda deveram-se ao trauma cirúrgico da castração, mas as diferenças entre os grupos foram associadas à capacidade do anti-inflamatório em controlar a inflamação. Em conclusão, baseado da resposta de fase aguda, o flunixin em comparação com o meloxicam e o firocoxib é mais eficiente no controle da inflamação após a castração em equinos.(AU)
Subject(s)
Animals , Male , Acute-Phase Proteins , Castration , Meloxicam , Horses/surgery , Anti-Inflammatory Agents/administration & dosage , Body Weight , OrchiectomyABSTRACT
Objective:To analysis the macrolide resistance, molecular characteristics and plused-field gel electrophoresis(PFGE) type of Bordetella pertussis ( Bp), explore the possible resistance mechanism and the relationship between PFGE types and macrolide resistance profiles. Methods:Erythromycin, azithromycin and clarithromycin susceptibility of clinical isolates during 2016 to 2018 was determined by E-test. PCR was used to detect the drug-resistant genes and mutation sites. PFGE were employed to do molecular typing for the strains.Results:Thirty-five strains were isolated, of which 27 strains were resistant to all three antibiotics, two strains were resistant to erythromycin and azithromycin, and six strains were sensitive to all three antibiotics. Partial macrolide resistant strains carried the methylase gene ermA (27.6%, 8/29) and ermB (31.0%, 9/29); A2047G site mutation was detected in macrolide-resistant strains, while no drug-resistant genes or mutation sites were found in sensitive strains. Resistant strains were classified into BPSR23 and BPFINR9 types, while sensitive strains were other profiles. Conclusions:The clinical isolated Bp were seriously resistant to erythromy and showed signs of resistance to other macrolides. The acquisition of methylase gene and mutation of A2047G site might be the main mechanism of resistance. The macrolide resistance might have has a certain correlation with PFGE profile.
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RESUMEN Objetivos: Determinar el efecto citotóxico y genotóxico in vitro del extracto crudo y etanólico del rizoma de Curcuma longa L. Materiales y métodos: El efecto citotóxico fue evaluado utilizando líneas celulares DU-145, HT-29, 3T3 BALB/c. Se hallaron los porcentajes de crecimiento en 48 horas y se determinó la concentración inhibitoria 50 (CI50). El efecto genotóxico en el ADN genómico humano se determinó mediante el método Tomasevich. Resultados: El extracto crudo produjo una CI50 de 12,98 ± 0,21 μg/mL para la línea celular tumoral HT-29, que es inferior a DU-145 con una CI50 de 36,77 ± 9,12 μg/mL; el extracto etanólico presentó una CI50 de 13,24 ± 0,77 y 20,54 ± 2,58 µg/mL para ambas líneas celulares, respectivamente; el compuesto estándar curcumina presentó una CI50 de 3,96 ± 0,60 y 13,94 ± 2,79 μg/mL, respectivamente. El extracto crudo a concentraciones de 50 y 100 mg/mL fragmentó entre el 40% a 95% de ADN genómico humano; mientras que, a 200 mg/mL, la fragmentación fue mayor al 95%. El extracto etanólico a todas las concentraciones no fragmentó el ADN. La curcumina a 200 mg/mL fragmentó menos del 5% de ADN genómico humano. Conclusiones: Los extractos crudo y etanólico de Curcuma longa L. demuestran efecto citotóxico in vitro diferencial para la línea celular tumoral humana DU-145 y HT29 semejante al compuesto estándar curcumina. El extracto crudo de Curcuma longa L. presenta una potente actividad genotóxica in vitro frente al ADN genómico humano, esta actividad está ausente en el extracto etanólico.
ABSTRACT Objectives: To determine the in vitro cytotoxic and genotoxic effect of the crude and ethanolic extract from the Curcuma longa L. rhizome. Materials and methods: The cytotoxic effect was evaluated using DU-145, HT-29, 3T3 BALB/c cell lines. The growth percentages in 48 hours; and the half maximal inhibitory concentration (IC50) were determined. The genotoxic effect on human genomic DNA was determined using the Tomasevich method. Results: Crude extract produced an IC50 of 12.98 ± 0.21 μg/mL for the HT-29 tumor cell line, which is lower than the value obtained for DU-145, with an IC50 of 36.77 ± 9.12 μg/mL. The ethanolic extract presented an IC50 of 13.24 ± 0.77 and 20.54 ± 2.58 μg/mL for both cell lines, respectively; the curcumin standard compound presented an IC50 of 3.96 ± 0.60 and 13.94 ± 2.79 μg/mL, respectively. Crude extract concentrations of 50 and 100 mg/mL fragmented between 40% to 95% of human genomic DNA; while at 200 mg/mL, fragmentation was greater than 95%. The ethanolic extract at all concentrations did not fragment the DNA. Curcumin at 200 mg/mL fragmented less than 5% of human genomic DNA. Conclusions: The crude and ethanolic extracts of Curcuma longa L. demonstrate different in vitro cytotoxic effects for the human tumor cell lines DU-145 and HT-29; similar to the standard curcumin compound. The crude extract of Curcuma longa L. shows a potent genotoxic in vitro activity against human genomic DNA; this type of effect is not produced by the ethanolic extract.
Subject(s)
In Vitro Techniques , Curcuma , Rhizome , Cell Line, Tumor , Complex Mixtures , Cell Line , HT29 Cells , Inhibitory Concentration 50 , BALB 3T3 CellsABSTRACT
ABSTRACT Irrigation water and cultivated soil have been identified as possible sources of contamination in several crops. In certain vegetables that are eaten raw, such as lettuce, this contamination can lead to public health problems. Aiming to evaluate the influence of these sources on the quality of lettuce grown in the Córrego Sujo Basin, Teresópolis, RJ, an important agricultural pole whose production services the metropolitan region of Rio de Janeiro, water from different sources (spring, weir and river) was collected in this region, as well as samples of soil and lettuce irrigated with these waters, to carry out conventional microbiological analyzes (counts of total heterotrophic bacteria and thermotolerant coliforms) and molecular analyzes (PCR-DGGE). The count of fecal coliforms in lettuce suggests that there is an influence of irrigation water and the cultivated soil on the contamination of these vegetables. The grouping of bacterial communities in the different samples obtained by the PCR-DGGE technique shows that irrigation water has a greater influence on the contamination of these vegetables in relation to the soil where they are grown. These results corroborate the need to monitor water bodies used for irrigation and demonstrate that the PCR-DGGE technique is of great value for the study of microbial communities and, when associated with specific primers, can help in the detection of pathogens in food.
RESUMO A água de irrigação e o solo de cultivo têm sido apontados como possíveis fontes de contaminação em diversas culturas. Em determinadas hortaliças consumidas cruas, como a alface, essa contaminação pode causar problemas de saúde pública. Objetivando avaliar a influência dessas fontes na qualidade das alfaces cultivadas na Bacia do Córrego Sujo, Teresópolis, RJ; importante polo agrícola com produção voltada à região metropolitana do Rio de Janeiro, coletou-se nesta região águas proveniente de diferentes fontes (nascente, açude e rio); solos e alfaces irrigados com essas águas, para realização de análises microbiológicas convencionais (contagens de bactérias heterotróficas totais e coliformes termotolerantes) e moleculares (PCR-DGGE). A contagem de coliformes fecais na alface sugere que existe influência da água de irrigação e do solo na contaminação desses vegetais. O agrupamento das comunidades bacterianas nas diferentes amostras obtido pela técnica de PCR-DGGE mostra que a água de irrigação tem influência maior na contaminação dessas hortaliças em relação ao solo onde são cultivadas. Esses resultados corroboram a necessidade de monitoramento de corpos d'água utilizados para irrigação e demonstram ser a técnica do PCR-DGGE de grande valia para o estudo das comunidades microbianas e, quando associada a iniciadores específicos podem ajudar na detecção de patógenos em alimentos.
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Aim: The present study was conducted to characterize VRSA isolates on the basis of pulsed field gel electrophoresis (PFGE) and the presence of spa gene, recovered from different Doon Valley Hospitals.Methodology: Six VRSA isolates were analyzed using PFGE and spa typing. spa gene coded Protein A was used as a genetic marker for the characterization of Staphylococcus aureus isolates. Dendrogram were constructed on the basis of unweighted pair group method with arithmetic means (UPGMA method) for clusters analyses.Results: Dendrogram finally showed two major banding patterns at about 85% similarity designated as PFGE type A and PFGE type B exhibiting differences of 4-6 bands. The length of spa gene varied from 1200 to 1500 bp, showing variation in length. The most prevalent length was 1200bp.
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Extracellular protease from culture supernatants of Streptomyces sp. LCJ12A was purified and characterized in thisstudy. The collected supernatant of Streptomyces sp. LCJ12A was purified by ammonium sulfate precipitation anddiethylaminoethyl cellulose chromatography. The molecular weight of the enzyme was 20.1 kDa. Sodium dodecylsulfate-polyacrylamide gel electrophoresis was used to determine the molecular weight of the enzyme. The culture filtrate,partially purified enzyme, and ammonium sulfate precipitate displayed catalysis and stability over pH 3–11.5 and showedoptimal activity at pH 10. The culture filtrate, ammonium sulfate precipitate, and partially purified enzyme were goodbetween 30 and 40°C and the optimum temperature was 35°C. The purified protease exhibited high catalytic activityand stability under different pH and temperature. The partially purified protease from Streptomyces sp. LCJ12A showeda substantial relative activity of 78% with bovine serum albumin (BSA), 43% with gelatin, and 96% relative activitywith casein. Lineweaver–Burk plot was used to calculate the Km and Vmax values for the partially purified protease.The Km values of Streptomyces sp. LCJ12A were 73.5 mM for casein, 44.54 mM for BSA, and 43.45 mM for gelatin.The Vmax values were 500 mM min−1 for casein, 303.03 mM min−1 for BSA, and 270.27 mM min−1 for gelatin fromStreptomyces sp. LCJ12A. The statistical analysis confirmed that compared to the other substrates, casein was significant.