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1.
Journal of Forensic Medicine ; (6): 763-773, 2022.
Article in English | WPRIM | ID: wpr-984169

ABSTRACT

In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.


Subject(s)
Forensic Medicine/methods , Body Fluids/chemistry , RNA/analysis , Feces , Forensic Genetics , Semen/chemistry , Saliva/chemistry
2.
Rev. chil. enferm. respir ; 37(3): 244-249, sept. 2021. tab
Article in Spanish | LILACS | ID: biblio-1388153

ABSTRACT

Resumen Los métodos diagnósticos clásicos para la tuberculosis son de baja sensibilidad o son muy lentos en la obtención de resultados (baciloscopía, cultivo de Koch). De ahí nace la necesidad de nuevos métodos diagnósticos para esta enfermedad. Los biomarcadores surgen como una opción a esta problemática, con un buen rendimiento diagnóstico, costo y accesibilidad. Ellos permiten identificar la respuesta inflamatoria y/o metabólica del huésped, extrapolando la presencia de Mycobacterium tuberculosis; o identifican moléculas propias del patógeno. En la presente revisión se describen biomarcadores que presentan un buen rendimiento diagnóstico basados en metodologías de investigación de alto nivel (estudio de cohortes, prospectivos, muestreo consecutivo o aleatorizado, comparación de rendimiento diagnóstico frente a cultivo). Es necesario el desarrollo de estas nuevas técnicas con el fin de realizar el diagnóstico precoz de la enfermedad y lograr así su tan ansiada eliminación.


The classical laboratory diagnostic methods for tuberculosis have a low sensitivity or take a long time to know their results. New methods are underway. Biomarkers are a good option to improve our diagnostic approach to this disease. They have good performance, low cost and accessibility. They identify a patient's inflammatory or metabolic response to Mycobacterium Tuberculosis or identifies molecules that are typical of the pathogen. In this paper we sum up the biomarkers with a good diag-nostic performance described in well design investigations. Early diagnosis with these new techniques should contribute to the elimination of the disease.


Subject(s)
Humans , Tuberculosis/diagnosis , Biomarkers/analysis , RNA/analysis , Proteins/analysis , Cytokines/analysis , Sensitivity and Specificity , Antibodies/analysis , Mycobacterium tuberculosis/isolation & purification , Mycolic Acids/analysis
3.
Braz. oral res. (Online) ; 34: e015, 2020. tab, graf
Article in English | LILACS | ID: biblio-1089381

ABSTRACT

Abstract We sought to compare the characteristics and clinical significance of neutrophil extracellular traps in gingival samples from patients with periodontitis and those with gingivitis. The clinical indexes of gingival samples from patients with periodontitis and gingivitis were measured; the expression of TNF-alpha and IL-8 was measured by real-time fluorescent quantitative PCR; and the expression of TLR-8 and MMP-9 was measured by western blotting assays. Chemotaxis, phagocytosis and phagocytic activity of neutrophils were measured. Compared with the healthy group, the expression of TNF-α and IL-8 in the periodontitis group and the gingivitis group increased significantly (p < 0.05), and TNF-α in the gingivitis group was significantly lower than that in the healthy group (p < 0.05). The expression of IL-8 in the periodontitis group was significantly higher than that in the periodontitis group (p < 0.05). Furthermore, the expression of TLR-8 and MMP-9 in the periodontitis group was different from that in the gingivitis group and the healthy group, and the expression of TLR-8 and MMP-9 in the gingivitis group was significantly different from that in the healthy group (p < 0.05). In addition, the neutrophil mobility index in healthy people was 3.02 ± 0.53, that in the periodontitis group was 2.21 ± 0.13, and that in the gingivitis group was 2.31 ± 0.12. In conclusion, the chemotaxis of neutrophils in gingival samples of patients with periodontitis and gingivitis was decreased, the phagocytotic ability and activity of neutrophils were reduced, and the release of the extracellular trap-releasing inducible factors TNF-alpha and IL-8 also declined.


Subject(s)
Humans , Male , Female , Young Adult , Periodontitis/pathology , Extracellular Traps , Gingivitis/pathology , Neutrophils/pathology , Reference Values , RNA/analysis , Case-Control Studies , Periodontal Index , Blotting, Western , Interleukin-8/analysis , Actins/analysis , Tumor Necrosis Factor-alpha/analysis , Matrix Metalloproteinase 9/analysis , Electrophoresis, Agar Gel , Toll-Like Receptor 8/analysis , Real-Time Polymerase Chain Reaction , Middle Aged
4.
Journal of Forensic Medicine ; (6): 387-392, 2019.
Article in English | WPRIM | ID: wpr-985022

ABSTRACT

Objective Quantitative analysis and comparison of the expression of ribonucleic acid (RNA) from frozen organs and formaldehyde-fixed and paraffin-embedded (FFPE) tissues. Methods Frozen specimens of human brain, myocardium and liver tissues as well as FFPE samples at different postmortem intervals were collected and mass concentration of RNA was extracted and detected. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technology was used to analyze the amplification efficiency and relative expression of each RNA marker. Results The mass concentration and integrity of RNA extracted from FFPE samples were relatively low compared with frozen specimens. The amplification efficiency of RNA markers was related with RNA species and the length of amplification products. Among them, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin (ACTB) with relatively long amplification products failed to achieve optimal amplification efficiency, whereas 5S ribosomal RNA (5S rRNA) achieved ideal amplification efficiency and showed quite stable expression across various tissues, therefore it was chosen as internal reference marker. The expression quantity of GAPDH and ACTB in frozen specimens with longer postmortem intervals and in FFPE samples with relatively long amplification products was decreased. The expressions of tissue-specific microRNAs (miRNAs), GAPDH and ACTB with relatively short amplification products had consistency in the same tissues and FFPE samples. Conclusion Through standardizing the RT-qPCR experiment, selecting the appropriate RNA marker and designing primers of appropriate product length, RNA expression levels of FFPE samples can be accurately quantified.


Subject(s)
Humans , DNA Primers , Formaldehyde , Gene Expression Profiling , MicroRNAs/analysis , Myocardium , Paraffin Embedding , RNA/analysis , Real-Time Polymerase Chain Reaction/standards
5.
Actual. osteol ; 13(3): 207-213, Sept - DIc. 2017. ilus, graf
Article in English | LILACS | ID: biblio-1117111

ABSTRACT

Osteocytes are the most abundant bone cell and are formed when osteoblasts become embedded in the bone matrix. Through changes in gene expression and paracrine effects, osteocytes regulate the number of osteoblasts, bone forming cells, and osteoclasts, bone resorbing cells, which are needed to maintain bone mass. MLO-Y4 is the better characterized osteocytic cell line; however, lacks expression of sclerostin, the product of the SOST gene, which is fundamental for osteocyte function and blocks bone formation. With the objective to isolate MLO-Y4 clones with different gene expression profiles, we performed cultures at very low density of MLO-Y4 cells stably transfected with nuclear green fluorescent protein (MLOnGFP). Cell morphology was visualized under a fluorescence microscope. Once the cells reached 80% confluency, RNA was extracted and quantitative real time PCR was performed. Clones exhibit different sizes and morphology, with some cells showing a spindle-like shape and others with abundant projections and a star-like shape. Gene expression also differed among clones. However, none of the clones examined expressed SOST. We conclude that the MLO-nGFP clones constitute a useful tool to study osteocyte differentiation and the role of osteocytes in the control of bone formation and resorption in vitro. (AU)


Los osteocitos son las células más abundantes del hueso y se forman cuando los osteoblastos se encuentran rodeados de matriz ósea. A través de cambios en la expresión génica y efectos paracrinos, los osteocitos controlan el número de osteoblastos que forman el hueso, y osteoclastos que resorben el hueso, células necesarias para mantener la masa ósea. Las células MLO-Y4 son la línea celular osteocítica más investigada; sin embargo, no expresan esclerostina, el pro esclerostina, el producto del gen SOST que bloquea la formación ósea y es indispensable para la función de los osteocitos. Con el objetivo de aislar clones de las células MLO-Y4 con diferentes perfiles de expresión génica, realizamos cultivos a muy baja densidad de las células transfectadas en forma estable con proteína verde fluorescente nuclear (MLO-nGFP). La morfología celular fue evaluada utilizando un microscopio de fluorescencia. Una vez que las células alcanzaron el 80% de confluencia, el ARN fue extraído y analizado por PCR cuantitativa en tiempo real. Las células de los diferentes clones tienen diferentes tamaños y morfología, algunas células son fusiformes y otras con proyecciones citoplasmáticas abundantes y en forma de estrella. La expresión de los genes también varió en los distintos clones. Sin embargo, ninguno de ellos expresó SOST. En conclusión, los clones de las células MLO-nGFP constituyen una herramienta útil para estudiar la diferenciación de los osteocitos y el rol de estas células en el control de la formación y resorción ósea in vitro. (AU)


Subject(s)
Humans , Male , Female , Osteoblasts/cytology , Osteoclasts/cytology , Osteocytes/cytology , Cell Line , Clone Cells/cytology , Osteoblasts/metabolism , Osteoclasts/metabolism , Osteocytes/metabolism , Osteogenesis/genetics , Bone Resorption/genetics , In Vitro Techniques , RNA/analysis , Gene Expression , Polymerase Chain Reaction , Collagen/genetics , Alkaline Phosphatase/metabolism , Fluorescence , Anti-Bacterial Agents/administration & dosage
6.
MedicalExpress (São Paulo, Online) ; 4(4)July-Aug. 2017. tab, graf
Article in English | LILACS | ID: biblio-894358

ABSTRACT

OBJECTIVES. The ANKRD1 gene codes for the ankyrin repeat domain containing protein 1 and has an important role in myogenesis and possibly also in angiogenesis. Microvasculopathy is a cornerstone and an early pathological marker of change in dermatomyositis, leading to hypoxia and muscle perifascicular atrophy. These alterations could upregulate genes involved in myogenesis and angiogenesis such as ANKRD1. Therefore, we analyzed ANKRD1 expression in muscle biopsies of dermatomyositis and correlated with other hypoxia parameters and with histological changes. METHODS. Total RNA was extracted from frozen muscle biopsies samples of 29 dermatomyositis patients. A control group consisted of 20 muscle biopsies from adult patients with non-inflammatory myopathy diseases. The gene coding for hypoxia-inducible factor 1, alpha subunit (HIF1A), was analyzed to estimate the degree of hypoxia. ANKRD1 and HIF1A transcript expression levels were determined by quantitative real time PCR. RESULTS. Significantly higher ANKRD1 and HIF1A expression levels were observed in dermatomyositis relative to the control group (P<0.001, both genes). In addition, ANKRD1 and HIF1A were coexpressed (r=0.703, P=0.001) and their expression levels correlated positively to perifascicular atrophy (r=0.420, P=0.023 and r=0.404, P=0.030, respectively). CONCLUSIONS. Our results demonstrate ANKRD1 overexpression in dermatomyositis correlated to HIF1A expression and perifascicular atrophy. ANKRD1 involvement in myogenesis and angiogenesis mechanisms indicates that further investigation is worthwhile.


OBJETIVOS: ANKRD1 codifica "ankyrin repeat domain containing protein 1" e tem um papel importante na miogênese e possivelmente também na angiogênese. Microvasculopatia é considerada como um ponto central e uma alteração patológica precoce na dermatomiosite (DM), levando à hipóxia e à atrofia perifascicular muscular. Estas alterações poderiam estimular genes envolvidos na miogênese e angiogênese como ANKRD1. Portanto, analisamos a expressão de ANKRD1 em biópsias musculares de DM e correlacionamos com outros parâmetros de hipóxia e alterações histológicas. MÉTODOS: O RNA total foi extraído de biópsias de músculos congelados de 29 pacientes com DM. Como grupo controle, foram usadas 20 biópsias de músculo de pacientes adultos com miopatia não-inflamatória. O gene que codifica a subunidade alfa do fator 1 induzido por hipóxia (HIF1A) foi analisado para estimar o grau de hipóxia. Os níveis de expressão dos transcritos ANKRD1 e HIF1A foram determinados por PCR quantitativa em tempo real. RESULTADOS: Níveis aumentados de expressões de ANKRD1 e HIF1A foram observados em DM quando comparados ao grupo controle (P<0,001, ambos os genes). Além disso, ANKRD1 e HIF1A apresentaram coexpressão (r=0,703, P=0,001) e seus níveis de expressão correlacionaram-se também positivamente com atrofia perifascicular (r=0,420, P=0,023 e r=0,404, P=0,030, respectivamente). CONCLUSÕES: Nossos resultados demonstraram aumento de expressão de ANKRD1 na DM, que correlacionou com a expressão de HIF1A e atrofia perifascicular. Investigações adicionais do envolvimento de ANKRD1 no mecanismo de miogênese e angiogênese devem ser realizadas.


Subject(s)
Humans , RNA/analysis , Muscle Development , Dermatomyositis/physiopathology , Hypoxia
7.
Electron. j. biotechnol ; 17(6): 287-295, Nov. 2014. ilus, graf, tab
Article in English | LILACS | ID: lil-730260

ABSTRACT

Background Abscisic acid (ABA)-, stress- and ripening-induced protein (ASR) is plant-specific hydrophilic transcriptional regulators involved in sucrose stress and wounding in banana. However, it is not known whether banana ASR genes confer salt stress tolerance. The contexts of the study was to analysis the sequence characterization of banana ASR1, and identify its expression patterns and function under salt stress using quantitative real-time PCR (qPCR) and overexpression in Arabidopsis. The purpose was to evaluate the role of banana ASR1 to salt stress tolerance employed by plants. Results A full-length cDNA isolated from banana fruit was named MaASR1, and it had a 432 bp open reading frame (ORF) encoding 143 amino acids. MaASR1 was preferential expression in roots and leaves compared to low expression in fruits, rhizomes and flowers. Under salt stress, the expression of MaASR1 quickly increased and highest expression level was detected in roots and leaves at 4 h, and then gradually decreased. These results suggested that MaASR1 expression was induced under salt stress. MaASR1 protein was localized in the nucleus and plasma membrane. MaASR1 was transformed to Arabidopsis and verified by southern and northern analysis, transgenic lines L14 and L38 integrated one and two copies of MaASR1, respectively, while overexpression in transgenic lines provided evidence for the role of MaASR1 to salt stress tolerance. Conclusions This study demonstrated that overexpression of MaASR1 in Arabidopsis confers salt stress tolerance by reducing the expression of ABA/stress-responsive genes, but does not affect the expression of the ABA-independent pathway and biosynthesis pathway genes.


Subject(s)
Plant Proteins/genetics , Plant Proteins/metabolism , Musa/genetics , Salt Tolerance , Plant Growth Regulators , RNA/analysis , Plants, Genetically Modified , Cloning, Molecular , Sequence Analysis , Arabidopsis , Abscisic Acid , DNA, Complementary/chemical synthesis , Real-Time Polymerase Chain Reaction , Salt Stress
8.
Electron. j. biotechnol ; 17(6): 251-261, Nov. 2014. ilus, graf, tab
Article in English | LILACS | ID: lil-730255

ABSTRACT

Background Fatty acid synthase (FAS) is a key enzyme of de novo lipogenesis (DNL), which has been cloned from several species: Gallus gallus, Mus musculus, Homo sapiens, but not from Anas platyrhynchos. The current study was conducted to obtain the full-length coding sequence of Peking duck FAS and investigate its expression during adipocyte differentiation. Results We have isolated a 7654 bp fragment from Peking duck adipocytes that corresponds to the FAS gene. The cloned fragment contains an open reading frame of 7545 bp, encodes a 2515 amino acid protein, and displays high nucleotide and amino acid homology to avian FAS orthologs. Twelve hour treatment of oleic acid significantly up-regulated the expression of FAS in duck preadipocytes (P < 0.05). However, 1000 µM treatment of oleic acid exhibited lipotoxic effect on cell viability (P < 0.05). In addition, during the first 24 h of duck adipocyte differentiation FAS was induced; however, after 24 h its expression level declined (P < 0.05). Conclusion We have successfully cloned and characterized Peking duck FAS. FAS was induced during adipocyte differentiation and by oleic acid treatment. These findings suggest that Peking duck FAS plays a similar role to mammalian FAS during adipocyte differentiation.


Subject(s)
Animals , Adipose Tissue/metabolism , Ducks , Fatty Acid Synthases/genetics , Fatty Acid Synthases/metabolism , Phylogeny , RNA/analysis , Gene Expression , Cell Differentiation , Cell Survival , Cloning, Molecular , Sequence Analysis , DNA, Complementary/chemical synthesis , Oleic Acid , Computational Biology , Lipogenesis
9.
Journal of Forensic Medicine ; (6): 368-374, 2013.
Article in English | WPRIM | ID: wpr-983854

ABSTRACT

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts. However, the conventional body fluid identification methods are prone to various limitations, such as time consumption, intensive labor, nonparallel manner, varying degrees of sensitivity and limited specificity. Recently, the analysis of cell-specific messenger RNA expression (mRNA profiling) has been proposed to supplant conventional methods for body fluid identification. Since 2011, the collaborative exercises have been organized by the European DNA Profiling Group (EDNAP) in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification. The major advantages of mRNA profiling, compared to the conventional methods, include higher sensitivity, greater specificity, the ability of detecting several body fluids in one multiplex reaction, and compatibility with current DNA extraction and analysis procedure. In the current review, we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.


Subject(s)
Humans , Blood Stains , Body Fluids/chemistry , DNA/analysis , DNA Primers , Forensic Medicine/methods , Gene Expression Profiling , RNA/analysis , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction/methods , Saliva/chemistry , Semen/chemistry
10.
The Korean Journal of Laboratory Medicine ; : 718-725, 2010.
Article in Korean | WPRIM | ID: wpr-73295

ABSTRACT

BACKGROUND: One of the major concerns with biobanking is the absence of standard operating procedures to eliminate pre-analytical variation arising from sample collection, preparation, and storage. Currently, there is a lack of tools to carry out quality control procedures for stored blood samples. The aim of this study is to assess the quality of stored blood samples in our biobank and to suggest appropriate indicators for their quality control. METHODS: The stored blood samples that we tested have been registered into our biobank since 2003. These were transferred to our biobank after carrying out routine requested tests, because the samples would have otherwise been discarded. For the purpose of quality control, we analyzed the concentrations and the integrity of DNA and RNA extracted from the stored samples and tested the levels of several serum proteins; the results were compared with the corresponding pre-storage levels. RESULTS: A total of 19 samples were stored from 2006 to 2009. Of the 22 samples stored between 2003 and 2005, 50% showed complete DNA integrity. However, sufficient RNA integrity was noted in only 1 sample stored as recently as 2009. High blood urea nitrogen levels were also noted in the stored sera, but the increase did not correlate to the duration of storage. CONCLUSIONS: The amount and integrity of nucleic acids extracted from stored blood samples are potential indicators that can be used for quality control. A guideline for the quality assessment of stored blood samples in a biobank is urgently needed.


Subject(s)
Blood Banks/standards , Blood Proteins/chemistry , Blood Urea Nitrogen , DNA/analysis , Clinical Laboratory Techniques , Quality Control , RNA/analysis , Specimen Handling/methods
11.
Article in English | IMSEAR | ID: sea-91254

ABSTRACT

The diagnosis of primary HIV-1 infection (PHI) is often missed and requires a high index of suspicion and a thorough knowledge of laboratory methods. We report the case of a young promiscuous male who presented with fever, rash and neurological symptoms 8 weeks after unprotected sexual exposure. Clinical and laboratory investigations showed the presence of leucopenia and thrombocytopenia with elevated transaminases, and a normal cerebrospinal fluid analysis, while CNS imaging revealed a vasculitis-like involvement of the corpus callosum. Symptoms resolved spontaneously over 3 weeks. Fourth generation ELISA with p24 antigen assay was positive with high HIV-1 RNA load while Western-Blot was negative, thus confirming the diagnosis of PHI. The patient was subsequently started on antiretroviral therapy (ART) and showed undetectable viral load after 8 weeks of therapy. We present the differential diagnoses which need to be entertained as well as the pros and cons of very early ART intervention.


Subject(s)
Adult , Anti-HIV Agents/therapeutic use , Antirheumatic Agents/therapeutic use , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay , HIV Infections/diagnosis , HIV-1/drug effects , Humans , Male , RNA/analysis
12.
Article in English | IMSEAR | ID: sea-45215

ABSTRACT

OBJECTIVE: Commercial TaqMan real-time PCR reagent was modified and applied on Light Cylcer 1.2 for quantifying HIV-1 RNA in plasma and compared with the reference method; COBAS AmpliPrep/COBAS Amplicor HIV-1 monitor test version 1.5. MATERIAL AND METHOD: Three hundred and eight frozen and fresh plasma samples were used for evaluation. Sequential specimens were also tested for follow-up cases. RESULTS: The correlation between HIV-1 RNA values obtained by reference and modified method with automated and manual sample preparation were significant with r = 0.916 and 0.908 (p < 0.001, p < 0.001) respectively with similar agreement log of mean bias (0.5 versus 0.48). High degree of correlation and agreement were observed between the assays in blind fresh plasma, r = 0.953 (p < 0.001) with 0.15 log difference in HIV-1 RNA level. Among follow-up samples, both methods gave 100% concordant results. CONCLUSION: This modified protocol provided evidence for using modified commercial real-time PCR reagent for HIV-1 RNA quantitative detection as a monitoring tool for HIV/AIDS patients in Thailand.


Subject(s)
HIV Infections/diagnosis , HIV-1 , Humans , Pilot Projects , Polymerase Chain Reaction/instrumentation , RNA/analysis , Reference Values , Thailand , Viral Load
13.
J Genet ; 2007 Aug; 86(2): 93-101
Article in English | IMSEAR | ID: sea-114314

ABSTRACT

A cytoplasmic male sterile (CMS) line of Brassica juncea was derived by repeated backcrossing of the somatic hybrid (Diplotaxis catholica + B. juncea) to B. juncea. The new CMS line is comparable to euplasmic lines for almost all characters, except for flowers which bear slender, needle-like anthers with aborted pollen. Detailed Southern analysis revealed two copies of coxI gene in the CMS line. One copy, coxI-1 is similar to the coxI gene of B. juncea, whereas the second copy, coxI-2 is present in a novel rearranged region. Northern analysis with eight mitochondrial gene probes showed altered transcript pattern only for the coxI gene. Two transcripts of 2.0 and 2.4 kb, respectively, were detected in the CMS line. The novel 2.4 kb transcript was present in floral bud tissue but absent in the leaf tissue. In plants where male sterility broke down under high temperature during the later part of the growing season, the 2.4 kb coxI transcript was absent, which suggested its association with the CMS. The two coxI genes from the CMS line showed two amino acid changes in the coding region. The novel coxI gene showed unique repeats in the 5' region suggesting recombination of mitochondrial genomes of the two species. The possible role of the duplicated coxI gene in causing male sterility is discussed.


Subject(s)
Base Sequence , Brassica/genetics , Cyclooxygenase 1/genetics , Cytoplasm/genetics , DNA, Mitochondrial/analysis , Flowers/genetics , Gene Duplication , Gene Expression , Genome, Plant , Hybrid Cells/metabolism , Molecular Sequence Data , Mustard Plant/genetics , Plant Infertility/genetics , RNA/analysis , Random Amplified Polymorphic DNA Technique , Sequence Homology, Nucleic Acid
15.
J Environ Biol ; 2002 Apr; 23(2): 143-6
Article in English | IMSEAR | ID: sea-113512

ABSTRACT

The effects of sublethal concentration of fenvalerate on DNA, RNA, RNA/DNA ratio and protein contents were estimated in gill and kidney tissues of an air breathing fish, Clarias batrachus. Fenvalerate reduced the DNA content in gill, whereas it did not produce any significant effect on DNA in kidney. This tissue-specific change in DNA content may be due to differential effects of fenvalerate or its metabolite(s) on synthesis and/degradation of DNA in gill and kidney cells of the fish. RNA and protein contents declined substantially in both the tissues in response to fenvalerate treatment. However, RNA/DNA ratio remains unchanged. It indicates that decrease in protein content in response to fenvalerate treatment might have been brought about by reduce rate of translation of messenger (mRNA) without a decrease in concentration of ribosomes.


Subject(s)
Animals , Catfishes/physiology , DNA/analysis , Dose-Response Relationship, Drug , Gills/chemistry , Insecticides/adverse effects , Kidney/chemistry , Nitriles , Pyrethrins/adverse effects , RNA/analysis
16.
J Environ Biol ; 2002 Jan; 23(1): 61-4
Article in English | IMSEAR | ID: sea-113263

ABSTRACT

Adult pre-spawning fish Labeo rohita were sublethally (1/5th 96h LC50) exposed to mercuric chloride and metacid-50 (methyl parathion). Accumulation of mercury and methyl parathion was studied and it was found that pre-spawning ovary appears as a potent organ for deposition of both the pollutants. RNA/DNA ratio of the control and treated fish were studied. It was found that the signifcant decrease in RNA/DNA ratio occurs after 9 and 30 days of exposure for mercury and 30 days for methyl parathion. Fluorescence microscopic studies by acridine orange staining method were also performed to show how much it is related to biochemical alterations. In some cases loss of metachromasia is correlated with the fall in RNA/DNA ratio. Some other abnormalities like fall in stage II: stage I oocyte ratio and necrosis was also observed.


Subject(s)
Animals , Carps/physiology , DNA/analysis , Drug Interactions , Environmental Pollutants/adverse effects , Female , Insecticides/adverse effects , Mercury/adverse effects , Methyl Parathion/adverse effects , Necrosis , Oogenesis/drug effects , Ovary/pathology , RNA/analysis
17.
Braz. j. microbiol ; 31(2): 77-82, Apr.-Jun. 2000. tab
Article in English | LILACS | ID: lil-297640

ABSTRACT

New and exciting molecular methods, many using the 16S small sub-unit ribosomal nucleic acid molecule, are opening the microbial "black box" in soil. These studies have added much to our knowlodge of microbial diversity in soils, and are beginning to advance our understanding of the relationship between this diversity and its function in soil processes. Over the next few years, the knowlodge gained from molecular studies will, we hope, lead to improvements in sustainable land management and sustainable exploitation of soil genetic resources. As we enter the third millenium, it is appropriate to review the application of 16S rDNA methods to soil microbiology. This review examines 16S ribosomal DNA (rDNA) methods and their application to soil. It mentions their limits and suggests how they may be applied in the future.


Subject(s)
DNA/analysis , Ecology , RNA/analysis , Soil Microbiology/standards , Genetics, Microbial/methods , Genetic Techniques/standards
18.
Medicina (B.Aires) ; 60(5/1): 587-90, 2000. tab
Article in English | LILACS, BINACIS | ID: lil-275469

ABSTRACT

The aim of this work to assess in the diversity of hepatitis C virus (HCV) quasispecies is related to histological severity and duration of infection in a cohort of untreated patients with an estimated onset of the disease. A total of 27 patients with diagnosis of chronic liver disease and history of blood transfusion (n = 16) or intravenous drug use (IDU) (n = 11) were included. All were anti-HCV positive and had detectable serum drug injection. Patients who consumed drugs for more than 2 years, or were coinfected with HBV or HIV were excluded. History of alcohol intake (> 80 g/day), ALT level and age at infection were recorded. Histological assessment of grading and staging was performed according to knodell score. The quasispecies diversity was investigated by single strand conformation polymorphism (SSCP) target to HVR-E2 region and SSCP pattern was evaluated as a single or multiple bands. The number of quasispecies did not correlate with estimated duration of the disease. Patients who acquired hepatitis C by blood transfusion did not differ in number of bands from patients who were IDU. There was no correlation between the heterogeneity of HCV quasispecies and age, serum ALT, Knodell score, HAI and fibrosis. In clonclusion the quasiespecies diversity of E2 had no correlation with grade and stage of chronic HCV infection and the presence of quasispecies was independent of the duration of the disease.


Subject(s)
Humans , Female , Adolescent , Adult , Middle Aged , Genetic Variation , Hepatitis C/virology , Hepacivirus/genetics , RNA/analysis , Polymerase Chain Reaction , Chronic Disease , Cohort Studies , Hepatitis C/immunology , Hepatitis C/pathology , Hepatitis C/transmission , Polymorphism, Single-Stranded Conformational , Liver/virology , Antibodies, Viral/analysis
19.
Acta cir. bras ; 13(1): 18-25, jan.-mar. 1998. graf
Article in English | LILACS | ID: lil-209226

ABSTRACT

Aiming at assessing compensatory lung growth after trilobectomy in rats, 3 groups of animals (control, thoracotomy and trilobectomy) were studied over 3 time intervals (7, 30 and 180 days post-operation). Protein. DNA and RNA contents in each lung were evaluated. The study of the left lung protein content reveals that compensatory growth ceased by day 30, whereas it continued to occur in the cranial lobe as long as 180 days post-operation. The lung DNA content in trilobectomized animals remained smaller than in the animals of the other groups demonstrating that compensatory growth was not brought about by hyperplasia. The lung RNA content in trilobectomized animals increased similarly to the lung protein content, demonstrating that the cells of the lung tissue must have had an increase in volume as no significant increase in their number occurred, as shown by the analysis of the lung DNA content. Therefore, it may be concluded that, in our experiment with adult animals, compensatory lung gowth after trilobectomy in rats occured due to an increase in the lung protein content and RNA content, suggesting a cellular volume increase (hypertrophy) and a probable increase in the intraveolar septs rather than an important cell multiplication.


Subject(s)
Animals , Male , Rats , DNA/analysis , Pneumonectomy , Proteins/analysis , Lung/physiology , Lung/chemistry , RNA/analysis , DNA/metabolism , Postoperative Period , Proteins/metabolism , Lung/growth & development , Rats, Wistar , RNA/metabolism , Time Factors
20.
Arch. med. res ; 28(4): 571-5, dec. 1997. ilus, tab
Article in English | LILACS | ID: lil-225265

ABSTRACT

Immunoprecipitation analysis of total HeLa cells RNA extract byprotein A-Sheparose purified autoantibodies and pCp 32P-3' end labeling RNAs revealed that U1, U2, U4 and U5 snRNAs are related with anti-Sm or U1nRNP autoantibodies, while the hY1, hY3, hY4 and hY5 scRNAs were related to anti-SSA/Ro autoantibodies present in sera of patient with Systemic Lupus Erythematosus. The authors detected molecular snRNAs and scRNAs specificities by autoantibodies in 71 sera, the molecular RNA specificity for anti-Sm (U1, U2, U4 and U5 snRNAs) was present in 39 percent; anti-SSA/Ro sera reacted against scRNAs (hY1, hY3, hY4 and hY5) in 36 percent, then anti-U1nRNP sera recognized U1 snRNA in 13 percent of sera and anti-rRNP related with rRNA were recognized in 8 percent. Twenty-nine SLE sera were RNA negative. A molecular characterization of the autoantibodies in sera from SLE patients may be a useful tool for clinical and laboratory diagnosis of SLE, and the use of autoantibodies es molecular probes allows to continue exploring some basic mechanism of gene expression


Subject(s)
Humans , RNA, Small Nuclear/analysis , RNA, Small Nuclear/immunology , Autoantibodies/blood , Autoantibodies/immunology , HeLa Cells , Lupus Erythematosus, Systemic/immunology , Lupus Erythematosus, Systemic/blood , Precipitin Tests , RNA/analysis , RNA/immunology
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