ABSTRACT
Objetivo: Demostrar la presencia de Rotavirus en las diferentes fases de un proceso de compostaje: matrices utilizadas como materia prima, mezcla a compostar y producto terminado. Materiales y métodos. El Rotavirus se determinó durante los tres procesos de compostaje. La detección viral se realizó por inmunocromatografía, ELISA y RT- PCR. Resultados. Se evidenció presencia viral en el primer proceso de compostaje, ausencia viral en el segundo y en el tercer proceso de compostaje, se presentaron interferencias que dificultaron interpretar los resultados de la PCR, lo cual impidió llegar a un resultado concluyente de su presencia en el abono. Conclusiones. Los abonos orgánicos pueden ser portadores de virus motivo por el cual se deben implementar pruebas de calidad para evitar que este material contribuya con la diseminación viral. Dentro de estos abonos existen sustancias capaces de interferir en las pruebas de detección.
Objective. To show the presence of rotavirus in different stages of a composting process: matrices used as raw material, mixture to be composted and the final product. Materials and methods. Immunochromatography, ELISA and RT-PCR were used for viral detection. Results. Rotavirus was found in the first composting step, no virus was found in the second step, and some inhibitory substances were found in the third step that posed difficulties in interpreting the PCR results and therefore providing a concluding result on rotavirus presence in the final product. Conclusions. Organic fertilizers can be vectors of human pathogenic viruses; therefore quality control tests must be implemented to avoid further viral dissemination. There are inhibitory substances present in organic fertilizers capable of interfering with the detection tests.
Objetivo. Demonstrar a presença de Rotavírus nas diferentes fases de um processo de compostagem: matrizes utilizadas como matéria-prima, mistura de composto e produto final. Materiais e métodos: A determinação do Rotavírus foi realizada nos três processos de compostagem. A detecção viral foi realizada por imunocromatografia, ELISA e RT-PCR. Resultados. Evidenciou-se a presença viral no primeiro processo de compostagem, ausência de vírus no segundo, e no terceiro processo de compostagem, apresentaram-se interferências que dificultaram a interpre tação dos resultados da PCR, tornando impossível chegar a um resultado conclusivo de sua presença no adubo. Conclusões. Os adubos orgânicos podem abrigar o vírus é por isso que se devem implementar provas de qualidade para evitar que esse material contribuía com a disseminação viral. Dentro desses fertilizantes existem substâncias capazes de interferir nas provas de detecção.
ABSTRACT
Compared with C3 plant,C4 plant had evident growth advantage,higher rates of water and nutrition using,and higher bio-yield than C3 plant,Sugarcane was one of the typical C4 plants.DNA was extracted from sugarcane leaves,primers were designed by the cDNA sequence of PPDK gene from GenBank.Then DNA was amplified by LA-PCR(Long Acute PCR) method,ligated into pMD18-T vectors,and transformed into E.coli.JM109.Full-length PPDK gene sequence of sugarcane was obtained,the sequence was 13.5kb in length.For convenient of the next experiment,two digest sites(XhoI and NotI) were introduced into primers,the full-length PPDK gene was splitted to two parts,and was ligated into pMD18-T simple vectors,respectively.Whole PPDK gene clone of sugarcane was finished.The strains were storaged in the lab.
ABSTRACT
Arkinson's disease(PD) is a neurodegenerative disease involving mainly the loss of dopaminergic neurons in substantia nigra by several factors. The cause of dopaminergic cell death is unknown. Recently, it has been focused on that Parkinson's disease resulting from mitochondrial dysfunction. In the previous studies, it was found that a 5 kilobase(kb) deletion derived from mtDNA dysfunction. And this result leads to a reduction of ATP production, which ultimately causes result in cell death. Blood samples were collected from 6 positive control(PC) and 9 PD patients. Total DNA was extracted twice with phenol followed by chloroform:isoamylalcohol(24: 1). For the analysis of mtDNA, polymerase chain reaction(PCR) and long and accurate polymerase chain reaction(LA PCR) were performed by mitochondrial specific primers. As a result, a deletions of large quantity was detected within several regions of mtDNA in PD patients. The analysis of the partial sequence of the mitochondrial D-loop gene and restriction fragment length polymorphism(RFLP) technique were performed to investigate the point mutation and nucleotide sequence variations between PC and PD patients. Fragment variations between PC and PD were seen in the fragment digested by Hin d III, Eco R V. These variations are attributed to the presence or absence of recognition site by base substitution. Point mutation was observed in the D-loop region. Patients 1 and 2 had one point mutation. Patient 1 had a transition from T to C at 195, and patient 2 had a transversion from A to T. In addition to point mutation, the deletion of mtDNA occurred complexI, III, IV and V subunits in PD patients.
Subject(s)
Humans , Adenosine Triphosphate , Base Sequence , Cell Death , DNA , DNA, Mitochondrial , Dopaminergic Neurons , Neurodegenerative Diseases , Parkinson Disease , Phenol , Point Mutation , Polymorphism, Restriction Fragment Length , Substantia NigraABSTRACT
Parkinson's disease(PD) is a neurodegenerative disorder characterised clinically by bradykinesia, rigidity, tremor, and pathologically by neuronal cell death in substantia nigra. The cause of dopaminergic neuronal cell death in PD remains unknown. Recently, decreased mitochondrial complex I activities have been reported in platelets, muscles, substantia nigra of the PD patients. Blood samples were lysed with lysis buffer, and incubated 1 hour with 20mg/ml proteinase K at 37degreesC. DNA was extracted with phenol and chloroform(1: ). The long and accurate polymerase chain reaction(LA PCR) was performed by mitochondrial specific primers. The mitochondrial ND1, ND2, CO I, CO II and 1/3ATPase 6/8, CO III, genes as well as parts of ND3 and 3/4ND5 subunit coding regions were analysed by LA PCR. In this study, it is observed not only 4,977 bp mtDNA deletion but a partial mtDNA deletion of the ND1, ND2, CO I~III genes in blood from patients with PD. The result of this study cannot rule out the possibility of point mutation. It is possible that such a deletion would cause mitochondrial dysfuncton, and as a result of mitochondrial dysfunction, Parkinson's disease could progress.