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1.
Chemosphere ; 308(Pt 3): 136569, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36155023

ABSTRACT

Green turtles, Chelonia mydas, have been included in biomonitoring efforts given its status as an endangered species. Many studies, however, rely on samples from stranded animals, raising the question of how death affects important biochemical and molecular biomarkers. The goal of this study was to investigate post mortem fluctuations in the antioxidant response and metabolism of carbohydrates in the liver of C. mydas. Liver samples were obtained from six green turtles which were submitted to rehabilitation and euthanized due to the impossibility of recovery. Samples were collected immediately after death (t = 0) and at various time intervals (1, 2, 3, 4, 5, 6, 12, 18 and 24 h post mortem), frozen in liquid nitrogen and stored at -80 °C. The activities of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G6PDH) were analyzed, as were the levels of lipid peroxidation, glycogen concentration, RNA integrity (RNA IQ) and transcript levels of carbonic anhydrase and pyruvate carboxylase genes. Comparison between post mortem intervals showed a temporal stability for all the biomarkers evaluated, suggesting that changes in biochemical and molecular parameters following green turtle death are not immediate, and metabolism may remain somewhat unaltered up to 24 h after death. Such stability may be associated with the overall lower metabolism of turtles, especially under an oxygen deprivation scenario such as organismal death. Overall, this study supports the use of biomarkers in sea turtles sampled within a period of 24 h post mortem for biomonitoring purposes, though it is recommended that post mortem fluctuations of particular biomarkers be evaluated prior to their application, given that proteins may show varying degrees of susceptibility to proteolysis.


Subject(s)
Carbonic Anhydrases , Turtles , Animals , Antioxidants/metabolism , Biomarkers/metabolism , Carbonic Anhydrases/metabolism , Catalase/metabolism , Glucosephosphate Dehydrogenase/genetics , Glucosephosphate Dehydrogenase/metabolism , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Glycogen/metabolism , Nitrogen/metabolism , Oxygen/metabolism , Pyruvate Carboxylase/metabolism , RNA/metabolism , Turtles/metabolism
2.
J Comp Pathol ; 157(1): 61-66, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28735673

ABSTRACT

The pathological and molecular findings associated with Talaromyces marneffei-induced pneumonia with concomitant infection by canine distemper virus (CDV) are described in a dog. The principal pathological alteration occurred in the lungs. Histopathology confirmed multifocal granulomatous pneumonia associated with numerous intralesional and intracellular septate fission cells consistent with T. marneffei. A molecular assay designed to amplify a partial fragment of the 18S rRNA gene of T. marneffei provided positive results from two fungal cultures derived from the lung. Sequencing and phylogenetic analyses confirmed the results of polymerase chain reaction (PCR). Furthermore, antigens of the CDV N protein were identified within the bronchial epithelium by immunohistochemistry and a PCR assay amplified the CDV N gene from hepatic and pulmonary fragments. Collectively, the pathological and molecular techniques confirmed a diagnosis of T. marneffei-induced pneumonia with concomitant infection by CDV. These findings represent the first description of pulmonary penicilliosis in the dog and extend the geographical niche of this emerging infectious pathogen. In this case, infection by CDV may have induced immunosuppression, which facilitated the development of pulmonary penicilliosis.


Subject(s)
Distemper/complications , Dog Diseases/microbiology , Mycoses/veterinary , Pneumonia/veterinary , Animals , Brazil , Dogs , Talaromyces
3.
Neotrop Entomol ; 46(1): 86-92, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27406593

ABSTRACT

The citrus fruit borer, Ecdytolopha aurantiana (Lima, 1927) (Lepidoptera: Tortricidae), is responsible for major losses to the citrus industry because it causes rot and drop of fruits. The current study aimed to select and characterize Bacillus thuringiensis (Berliner, 1911) strains toxic to E. aurantiana. For this purpose, 47 B. thuringiensis strains were evaluated in selective bioassays using first instar larvae of E. aurantiana. The lethal concentration (LC50) of the most toxic strains was estimated, and the strains were characterized by morphological, biochemical, and molecular methods. Of the 47 strains tested, 10 caused mortality above 85% and showed mean lethal concentrations between 1.05E+7 and 1.54E+8 spores mL-1. The lowest LC50 values were obtained for the HD-1 standard strain and the BR145, BR83, BR52, and BR09 strains. The protein profile showed the presence of Cry proteins of 60, 65, 70, 80, and 130 kDa. The molecular characterization showed the presence of cry1, cry2, cry3, and cry11 genes. The morphological analysis identified three different crystalline inclusions: bipyramidal, round, and cuboidal. The cry1 and cry2 genes were the most frequent among the B. thuringiensis strains evaluated and encode Cry proteins toxic to insects of the order Lepidoptera, which agree with the toxicity results obtained by the selective bioassays against E. aurantiana. The results showed four different B. thuringiensis strains toxic to E. aurantiana at the same level as the HD-1 standard strain, and these strains have biotechnological potential for E. aurantiana control through the production of transgenic plants or the formulation of biopesticides.


Subject(s)
Bacillus thuringiensis , Lepidoptera , Pest Control, Biological , Animals , Bacillaceae , Bacterial Proteins , Endotoxins , Hemolysin Proteins
4.
Braz J Microbiol ; 44(1): 37-41, 2013.
Article in English | MEDLINE | ID: mdl-24159281

ABSTRACT

The presence of Salmonella in the intestinal tract, on the chickens skin and among their feathers, may cause carcasses contamination during slaughtering and processing and possibly it is responsible by the introduction of this microorganism in the slaughterhouses. A rapid method to identify and monitor Salmonella and their sorovars in farm is becoming necessary. A pre-enriched multiplex polymerase chain reaction (m-PCR) assay employing specific primers was developed and used to detect Salmonella at the genus level and to identify the Salmonella enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica serovar Typhimurium (S. Typhimurium) in broiler chicken swab samples. The method was validated by testing DNA extract from 90 fresh culture cloacal swab samples from poultry chicken cultured in phosphate buffer peptone water at 37 °C for 18 h. The final results showed the presence of Salmonella spp. in 25% of samples, S. Enteritidis was present in 12% of the Salmonella-positive samples and S. Typhimurium in 3% of the samples. The m-PCR assay developed in this study is a specific and rapid alternative method for the identification of Salmonella spp. and allowed the observation of specific serovar contamination in the field conditions within the locations where these chickens are typically raised.

5.
Transpl Infect Dis ; 15(1): 42-8, 2013 Feb.
Article in English | MEDLINE | ID: mdl-22783905

ABSTRACT

BACKGROUND: Respiratory syncytial virus (RSV) is a common cause of seasonal respiratory viral infection in hematopoietic stem cell transplantations (HSCT) patients. The efficacy of treatment, however, remains controversial. We describe an outbreak of 31 cases of RSV that occurred in an HSCT outpatient care unit in the fall season from March through May 2010, with a good outcome without any specific antiviral treatment. METHODS: During these 3 months, 222 nasal wash samples were tested and, of these, 31 outpatients were positive for RSV. In 2009, 99 samples had been tested and only 10 outpatients were positive for RSV in the same period. RESULTS: Seven (22.5%) patients had severe neutropenia (<500 cells/µL); severe lymphopenia (<200 cells/µL) was present in 13 (41.9%) patients, and 14 (45%) had received intravenous broad-spectrum antibiotics. Hospitalization was necessary only for 8 patients (25.8%); 20 had lower respiratory tract infection (64.5%). Only 1 patient died as a result of proven invasive aspergillosis. CONCLUSION: This report suggests that HSCT outpatients with no risk factors may not always require specific treatment for RSV.


Subject(s)
Antiviral Agents/therapeutic use , Hematopoietic Stem Cell Transplantation , Outpatients , Respiratory Syncytial Virus Infections/epidemiology , Adolescent , Adult , Aged , Child , Cross Infection , Disease Outbreaks , Female , Humans , Immunocompromised Host , Male , Middle Aged , Nasal Lavage Fluid/virology , Respiratory Syncytial Virus Infections/drug therapy , Respiratory Syncytial Viruses/isolation & purification , Risk Factors , Treatment Outcome , Young Adult
6.
Braz. j. microbiol ; 44(1): 37-42, 2013. ilus
Article in English | LILACS | ID: lil-676883

ABSTRACT

The presence of Salmonella in the intestinal tract, on the chickens skin and among their feathers, may cause carcasses contamination during slaughtering and processing and possibly it is responsible by the introduction of this microorganism in the slaughterhouses. A rapid method to identify and monitor Salmonella and their sorovars in farm is becoming necessary. A pre-enriched multiplex polymerase chain reaction (m-PCR) assay employing specific primers was developed and used to detect Salmonella at the genus level and to identify the Salmonella enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica serovar Typhimurium (S. Typhimurium) in broiler chicken swab samples. The method was validated by testing DNA extract from 90 fresh culture cloacal swab samples from poultry chicken cultured in phosphate buffer peptone water at 37 ºC for 18 h. The final results showed the presence of Salmonella spp. in 25% of samples, S. Enteritidis was present in 12% of the Salmonella-positive samples and S. Typhimurium in 3% of the samples. The m-PCR assay developed in this study is a specific and rapid alternative method for the identification of Salmonella spp. and allowed the observation of specific serovar contamination in the field conditions within the locations where these chickens are typically raised.


Subject(s)
Humans , Animals , Poultry/analysis , Food Microbiology , In Vitro Techniques , Polymerase Chain Reaction , Polymerase Chain Reaction/methods , Salmonella Infections, Animal , Skin , Salmonella enteritidis/isolation & purification , Salmonella typhimurium/isolation & purification , Food Samples , Methods , Serotyping
7.
Transpl Infect Dis ; 14(6): 564-74, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22882692

ABSTRACT

BACKGROUND: The 2009 pandemic influenza A (H1N1) virus spread rapidly throughout Brazil. Non-adjuvanted and the adjuvanted influenza A H1N1/09 monovalent vaccine were recommended as a single dose to persons at risk including renal transplant recipients (RTR). We analyzed the safety and the immune response of 2 influenza A H1N1/09 monovalent vaccines in RTR, and identified factors influencing the immune response. METHODS: A total of 78 RTR received a single dose of either influenza A H1N1 2009 monovalent AS03-adjuvanted vaccine or a non-adjuvanted vaccine, and 58 healthy controls received a single dose of non-adjuvanted vaccine. Antibody responses to influenza A H1N1 were measured by hemagglutination inhibition assay and were compared between groups on the day of vaccination and 21-30 days thereafter, using geometric mean titer (GMT), and seroprotection (SP) and seroconversion (SC) rates. RESULTS: Among RTR, after adjuvanted and non-adjuvanted H1N1 vaccination, the SP rate increased from 16.7% to 61.7% (P < 0.001) and to 50% (P < 0.001), and SC rates were 61.7% and 50%, respectively. For healthy controls, SP rate increased from 25.8% to 89.7% (P < 0.001), and SC rate was 87.9% after vaccination. Pre-vaccination GMT for the adjuvanted and non-adjuvanted RTR vaccine groups and healthy controls was 9.7 (95% confidence interval [CI] 7.3-13.1), 8.9 (95% CI 5.4-14.7), and 12.5 (95% CI8.7-18.2), and significantly increased to 49.8 (95% CI 31.3-79.4, P < 0.001), 43.2 (95% CI 16.3-114.4, P < 0.001), and 323.8 (95% CI 213.9-490.2, P < 0.001), respectively. Deceased-donor type transplant significantly reduced SP (odds ratio [OR] = 4.62, 95% CI 1.36-15.69, P = 0.014) and SC (OR = 6.29, 95% CI 1.89-20.98, P = 0.003) rates, and younger age positively affected SP (OR = 0.11; 95% CI 0.03-0.04, P = 0.001). Adverse events were mild, and renal function showed no change post vaccination. CONCLUSION: RTR vaccinated with either an adjuvanted or non-adjuvanted monovalent influenza vaccine presented poor response compared with healthy controls. Post-vaccination adverse events were mild, and no rejection episode or renal dysfunction was observed.


Subject(s)
Antibodies, Viral/blood , Influenza A Virus, H1N1 Subtype/immunology , Influenza Vaccines/immunology , Influenza, Human/prevention & control , Kidney Transplantation/immunology , Adjuvants, Anesthesia , Brazil/epidemiology , Humans , Influenza, Human/epidemiology , Influenza, Human/virology , Pandemics
8.
J Agric Food Chem ; 59(12): 6568-74, 2011 Jun 22.
Article in English | MEDLINE | ID: mdl-21574572

ABSTRACT

The production of volatile compounds by microbial communities of cork samples taken during the cork manufacturing process was investigated. The majority of volatiles were found in samples collected at two stages: resting after the first boiling and nontreated cork disks. Volatile profiles produced by microbiota in both stages are similar. The releasable volatile compounds and 2,4,6-trichloroanisole (TCA) produced in cork-based culture medium by five isolated fungal species in pure and mixed cultures were also analyzed by gas chromatography coupled with mass spectrometry (GC-MS).The results showed that 1-octen-3-ol and esters of fatty acids (medium chain length C8-C20) were the main volatile compounds produced by either pure fungal species or their mixture. Apparently, Penicillium glabrum is the main contributor to the overall volatile composition observed in the mixed culture. The production of releasable TCA on cork cannot be attributed to any of the assayed fungal isolates.


Subject(s)
Fungi/metabolism , Quercus/chemistry , Quercus/microbiology , Volatile Organic Compounds/metabolism , Anisoles/analysis , Fungi/isolation & purification , Volatile Organic Compounds/analysis
9.
Transplant Proc ; 41(3): 962-4, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19376399

ABSTRACT

OBJECTIVE: Arrhythmogenic right ventricular dysplasia (ARVD) is a myocardial disease of familiar, origin where the myocardium is replaced by fibrofatty tissue predominantly in the right ventricle. Herein we have presented the clinical courses of 4 patients with ARVD who underwent orthotopic heart transplantation. PATIENTS AND METHODS: Among 358 adult patients undergoing heart transplantation, 4 (1.1%) displayed ARVD. The main indication for transplantation was the progression to heart failure associated with arrhythmias. All 4 patients displayed rapid, severe courses leading to heart failure with left ventricular involvement and uncontrolled arrhythmias. RESULTS: In all cases the transplantation was performed using a bicaval technique with prophylactic tricuspid valve annuloplasty. One patient developed hyperacute rejection and infection, leading to death on the 7th day after surgery. The other 3 cases showed a good evolution with clinical remission of the symptoms. Pathological study of the explanted hearts confirmed the presence of the disease. CONCLUSIONS: ARVD is a serious cardiomyopathy that can develop malignant arrhythmias, severe ventricular dysfunction with right ventricular predominance, and sudden cardiac death. Orthotopic heart transplantation must always be considered in advanced cases of ARVD with malignant arrhythmias or refractory congestive heart failure with or without uncontrolled arrhythmias, because it is the only way to remit the symptoms and the disease.


Subject(s)
Arrhythmias, Cardiac/etiology , Arrhythmias, Cardiac/surgery , Heart Transplantation/methods , Ventricular Dysfunction, Right/surgery , Adolescent , Cardiomyopathies/etiology , Cardiomyopathies/genetics , Cardiomyopathies/surgery , Chromosomes, Human , Electrocardiography , Female , Genes, Dominant , Heart Transplantation/mortality , Humans , Male , Young Adult
10.
Vet Parasitol ; 160(3-4): 225-9, 2009 Mar 23.
Article in English | MEDLINE | ID: mdl-19091475

ABSTRACT

The objective of this work was to carry out a study on caprine toxoplasmosis in the state of Minas Gerais, Brazil. To determine the prevalence of toxoplasmosis in goats in Minas Gerais, 767 sera from goats were tested by ELISA (enzyme-linked immunosorbent assay) and IFAT (indirect fluorescence antibody test). The prevalence of antibodies to Toxoplasma gondii was 43.0% and 46.0% by ELISA and IFAT, respectively. It was observed that 26.8% of the goats show low-avidity IgG to T. gondii. These results suggest the presence of animals in recent phase of toxoplasmosis in Minas Gerais. The risk factors for toxoplasmosis in goats were: age over 36 months (OR=1.21; IC 95% 1.02-1.44), use of pen (OR=1.83; IC 95%1.01-3.31) and pure breed animals (OR=2.49; IC 95% 1.11-5.59).


Subject(s)
Antibodies, Protozoan/blood , Goat Diseases/epidemiology , Toxoplasma/immunology , Toxoplasmosis, Animal/epidemiology , Age Factors , Animals , Brazil/epidemiology , Breeding , Confidence Intervals , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Fluorescent Antibody Technique, Indirect/methods , Fluorescent Antibody Technique, Indirect/veterinary , Goats , Immunoglobulin G/blood , Male , Odds Ratio , Risk Factors , Seroepidemiologic Studies
11.
Bone Marrow Transplant ; 38(1): 37-40, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16715111

ABSTRACT

Reimmunization guidelines have recommended the inactivated HAV vaccine for hematopoietic stem cell transplant (HSCT) recipients living in or traveling to areas where hepatitis A is endemic. As a shift from high to medium hepatitis A endemicity has been observed in several countries in Latin America, we conducted a retrospective study to evaluate the prevalence of hepatitis A pre-bone marrow transplant (BMT) and the loss of specific antibodies in consecutive stored serum samples from 77 BMT recipients followed up from 82 to 1530 days. The prevalence of HAV antibodies was 92.2% before BMT. As vaccine was not available in Brazil when the samples were taken, it was assumed that this prevalence reflects natural infection. Survival analysis showed that the probability of becoming seronegative was 4.5% (+/-2.6%), 7.9% (+/-3.4%), 10.1% (+/-4.0%), 23.4% (+/-9.6%) at 1, 2, 3 and 4 years after transplant, respectively. The loss of HAV antibodies was significantly associated with longer follow-up (P=0.0015), younger age (P=0.049) and acute graft-versus-host disease (P=0.035). As most reimmunization protocols start around day +365, in developing countries with similar HAV endemicity, BMT recipients should have serological screening before HAV vaccination and the inactivated vaccine should be advised to those seronegative.


Subject(s)
Antibody Formation , Bone Marrow Transplantation/adverse effects , Hepatitis A Antibodies/blood , Adolescent , Adult , Antibody Formation/immunology , Brazil/epidemiology , Child , Child, Preschool , Follow-Up Studies , Hepatitis A/blood , Hepatitis A/epidemiology , Humans , Middle Aged , Prevalence , Retrospective Studies , Risk Factors , Seroepidemiologic Studies , Serologic Tests , Survival Analysis
12.
J Hered ; 96(5): 600-2, 2005.
Article in English | MEDLINE | ID: mdl-15994414

ABSTRACT

The giant anteater (Myrmecophaga tridactyla) is found from Belize and Guatemala to Paraguay and Argentina. Its conservation status is considered vulnerable by IUCN. Here we report the isolation and characterization of six microsatellite loci. Positive loci for (GT)(n) were isolated using a magnetic bead hybridization selection protocol. The number of alleles per locus as well as the heterozygosity and PCR conditions are described. These loci will be useful for studying population structure, genetic diversity, and paternity in M. tridactyla wild populations.


Subject(s)
Microsatellite Repeats/genetics , Xenarthra/genetics , Animals , Base Sequence , DNA Primers , Heterozygote , Molecular Sequence Data , Sequence Analysis, DNA
13.
J Autoimmun ; 17(3): 223-8, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11712860

ABSTRACT

The detection of anti-actin (AAA) by immunofluorescence is hindered by the presence of a serum factor. To better understand how it interferes with AAA detection, we tested sera from 20 patients with autoimmune hepatitis, and from 21 healthy adults, diluted 1:10 and prepared as follows: (A) diluted with PBS; (B) inactivated at 56 degrees C, and diluted with PBS; (C) diluted with 34 mM EDTA/PBS; (D) heated and diluted with EDTA/PBS. To reveal AAA, a fluorescein-labelled anti-human IgG was used in the process of indirect immunofluorescence. In a parallel assay, the substrate, acetone-fixed human fibroblasts, was preincubated with sera prepared as if it were to identify AAA, but instead, a rhodamine-phalloidin was used to identify F-actin, by direct immunofluorescence. All sera from patients were reactive to AAA when heat-inactivated and/or calcium-chelated, and 60% of them when diluted with unmodified sera (P=0.004). F-actin continued to be present after preincubation with heat-inactivated or calcium-chelated sera from patients and healthy controls, and in 41.5% of reactions with unmodified serum (P=0.0000001). The heat inactivation and the calcium chelation were both efficient procedures for maintaining the microfilament structure intact after serum incubation and, therefore, for identifying AAA.


Subject(s)
Actins/immunology , Actins/metabolism , Autoantibodies/blood , Calcium/physiology , Hot Temperature , Microfilament Proteins/physiology , Actin Depolymerizing Factors , Actins/blood , Adult , Autoantibodies/analysis , Calcium/blood , Destrin , Fluorescent Antibody Technique , Hepatitis, Autoimmune/blood , Hepatitis, Autoimmune/immunology , Humans , Microfilament Proteins/blood , Polymers/metabolism
14.
Transplantation ; 71(3): 412-7, 2001 Feb 15.
Article in English | MEDLINE | ID: mdl-11233903

ABSTRACT

BACKGROUND: Antigenemia and quantitative polymerase chain reaction (PCR) are widely used for cytomegalovirus (CMV) diagnosis after heart transplantation due to their enhanced predictive values for disease detection when specific cut-off values are used. The purpose of this study was to compare, in the same patient setting, the predictive values of quantitative PCR and antigenemia for CMV disease detection, using specific cut-off values. METHODS: Thirty heart transplant receptors were ch prospectively monitored for active CMV infection and disease detection, using quantitative PCR and anti- po genemia. Positive and negative predictive values for pr CMV disease detection were calculated using cut-off pr values for both antigenemia (5 and 10 positive cells/300,000 neutrophils) and quantitative-PCR (50,000 and 100,000 copies/10(6) leukocytes). RESULTS: Active CMV infection was diagnosed in 93.3% of patients and CMV disease in 23.3%. The positive and negative predictive (%) values for CMV disease detection were 35/100 and 46.7/100, respectively, for quantitative PCR and antigenemia. Using 5 and 10 positive cells/300,000 neutrophils as cut-off values for antigenemia, the positive and negative predictive values (%) for disease detection were respectively 63.6/100 and 70/100. For quantitative PCR, the positive and th negative predictive values (%) for cut-off values of to 50,000 and 100,000 copies/10(6) leukocytes were 53.8/100 and 60/94.1, respectively. CONCLUSION: In our series, antigenemia and quantitative-PCR had enhanced and similar predictive values for CMV disease detection when specific cut-off values were used. The choice between these two methods for disease detection may rely less on their efficiency and more on the experience and familiarity with them.


Subject(s)
Antigens, Viral/blood , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/immunology , Heart Transplantation/adverse effects , Polymerase Chain Reaction/methods , Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Time Factors
15.
J Chromatogr A ; 889(1-2): 51-7, 2000 Aug 11.
Article in English | MEDLINE | ID: mdl-10985535

ABSTRACT

The adsorption and desorption capacities of 11 different solid-phase extraction sorbents were tested for the preconcenration of pigments of various Hungarian red wines. The concentrates were evaluated by multiwavelengh spectrophotometry combined with a spectral mapping technique (SPM) and by reversed-phase high-performance liquid chromatography. The highest (10-fold) concentration of pigments was achieved on octadecylsilica sorbent. It can be used five times without losing adsorption and desorption characteristics. SPM indicated that multiwavelength spectrophotometry can be employed for the differentiation of red wines. Comparison of the chromatograms of pigments with and without preconcentration showed that preconcentration makes possible the separation and detection of pigments present in low concentration in red wines.


Subject(s)
Chromatography, High Pressure Liquid/methods , Pigments, Biological/isolation & purification , Wine/analysis , Spectrum Analysis , Wine/classification
16.
J Chromatogr Sci ; 38(4): 145-50, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10766480

ABSTRACT

The soluble color pigments of raisin are separated by reversed-phase thin-layer chromatography (TLC), and the capacity of TLC-Fourier transform infrared (FTIR) with both on-line and off-line coupling is assessed for the identification of the main fraction. TLC has also been used as a pilot technique for the development of a gradient elution method for the separation of pigments by high-performance liquid chromatography (HPLC). On-line TLC-FTIR cannot be used for identification because of the strong adsorbance of the stationary phase. Off-line TLC-FTIR combined with the retention behavior of the main pigment fraction indicates that it is a polymer, caramel-like compound composed of erythrose and fructose monomers. Baseline separation of pigments is achieved by HPLC using TLC as a pilot method.


Subject(s)
Chromatography, High Pressure Liquid/methods , Chromatography, Thin Layer/methods , Fruit/chemistry , Reproducibility of Results , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared
17.
Hepatology ; 23(5): 1098-104, 1996 May.
Article in English | MEDLINE | ID: mdl-8621140

ABSTRACT

In autoimmune hepatitis (AIH), the smooth-muscle antibody is specific for polymerized actin. Detection of antiactin antibody (AAA) has been hampered by technical problems. We have investigated AAA in 30 sera from patients with liver diseases and smooth-muscle antibody. AAA was detected by indirect immunofluorescence in 1:40, 1:80, and 1:160 dilutions. Five techniques were performed using fibroblasts: with vinblastine (A); without drugs (B); with sodium citrate (C); without drugs but with heat serum inactivation (D); and with sodium citrate and heat serum inactivation (E). For comparative analysis, we considered: the total number of AAA-positive sera regardless of the dilution in which reactivity was observed, as well as in each dilution separately; and the comparison of AAA intensity between 1:40 x 1:80, 1:40 x 1:160, and 1:80 x 1:160 dilutions. AAA was more positive in techniques B, C, D, and E than in A (P < .001) in general, and in each dilution separately. AAA was more positive in technique D than in B in 1:40 (P = .0005) and 1:80 dilutions (P = .03), as well as in E than in C (P = .0001) in 1:40 dilution. Techniques B and D yielded results similar to C and E, respectively. AAA staining was significantly more intense in 1:80 and 1:160 than in 1:40 dilution in A, B, and C; it was both significantly less intense in 1:80 and 1:160 than in 1:40 dilution and in 1:80 than in 1:160 in techniques D and E. We concluded that heat inactivation increased AAA seropositivity/intensity in 1:40 and 1:80 dilutions, preventing false-negative results; actin polymerization with sodium citrate did not enhanced AAA seropositivity/intensity. The technique with vinblastine was the least effective.


Subject(s)
Actins/immunology , Actins/metabolism , Autoantibodies/blood , Autoantibodies/analysis , Cells, Cultured , Citrates , Citric Acid , Female , Fibroblasts/immunology , Fluorescent Antibody Technique, Indirect , Hot Temperature , Humans , Liver Diseases/immunology , Male , Muscle, Smooth/immunology , Polymers , Vinblastine
18.
Rev Inst Med Trop Sao Paulo ; 38(3): 165-9, 1996.
Article in English | MEDLINE | ID: mdl-9163979

ABSTRACT

The prevalence of antibodies against cytomegalovirus (CMV) and the incidence of CMV infection were tested in 98 children aged 5 to 36 months who attended the day-care center of a University hospital in São Paulo. At the beginning of the study the overall prevalence of anti-CMV IgG antibodies was 44% (43/98). Saliva and/or urine samples were obtained from 38 of the 43 children that were seropositive at the beginning of the study for isolation of the virus, and 52.6% of these children were found to excrete CMV in one of the two materials. Among the 37 children that were initially seronegative from whom it was possible to obtain a new blood sample 6 to 12 months later, 22 (59.5%) presented seroconversion. The rate of viral excretion through urine or saliva from the children that seroconverted was 50%. These results indicate that CMV infection is frequent and occurs early among the children who attend this day-care center. However, controlled studies using molecular epidemiology techniques are needed to define more precisely the role of day-care centers in CMV dissemination.


Subject(s)
Antibodies, Viral/isolation & purification , Child Day Care Centers , Cytomegalovirus Infections/epidemiology , Cytomegalovirus/immunology , Brazil , Child, Preschool , Cytomegalovirus/isolation & purification , Cytomegalovirus Infections/blood , Female , Humans , Incidence , Infant , Male , Prevalence
19.
J Chemother ; 7(2): 126-32, 1995 Apr.
Article in English | MEDLINE | ID: mdl-7666118

ABSTRACT

Simultaneous administration of antacids containing magnesium or aluminium and ciprofloxacin or other quinolones decreases the gastrointestinal absorption of those antibacterial agents. Current speculation about the mechanism of this interaction has focused on drug-cation chelation. The present study was designed to detect the protonation in solutions and the formation of the complex species at the pH levels typical of the gastrointestinal tract. It involves the study of ciprofloxacin in aqueous solutions containing Al3+ and (or) Mg2+ by combining the results of potentiometric and spectroscopic (1H nuclear magnetic resonance) techniques. Calculations were only performed for data in the range 4.5 < pH < 5.5 (pH levels typical of gastrointestinal tract) and the results of both methods are made self-consistent, assuming an equilibrium model including complex species MHL, MLOH (where H2L denotes ciprofloxacin and M is Al3+ or Mg2+); their formation constants are given.


Subject(s)
Aluminum/chemistry , Aluminum/metabolism , Antacids/chemistry , Antacids/metabolism , Ciprofloxacin/chemistry , Ciprofloxacin/metabolism , Magnesium/chemistry , Magnesium/metabolism , Antacids/pharmacology , Cations/chemistry , Ciprofloxacin/pharmacology , Drug Interactions , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , Potentiometry
20.
Rev. Inst. Med. Trop. Säo Paulo ; 33(4): 243-50, jul.-ago. 1991. tab
Article in Portuguese | LILACS | ID: lil-108389

ABSTRACT

Com o objetivo de determinar a prevalencia da infeccao pelo Citomegalovirus (CMV) em pacientes com AIDS, bem como relacionar os achados clinicos virologicos decorrentes desta infeccao com as repercussoes anatomopatologicas, estudamos 50 pacientes adultos atendidos entre abril de 1986 a junho de 1987, em dois hospitais publicos de Sao Paulo (HSP e HSPE). Estes pacientes foram acompanhados clinica e laboratorialmente, por periodo medio de 2 meses com coletas seriadas de sangue, urina e saliva. Foram realizados isolamento do CMV em monocamadas de fibroblastos humanos e testes sorologicos de Imunofluorescencia Indireta (IFI-IgG/IgM) e Reacao Imunoenzimatica (ELISA-IgG). No momento da admissao no estudo 20 por cento (10/50) dos pacientes apresentavam anticorpos IgM CMV especificos e 100 por cento (50/50) deles anticorpos IgG (IFI). Durante o acompanhamento, 5 pacientes inicialmente IgM negativos tornaram-se IgM positivos, sugerindo reativacao ou reinfeccao pelo CMV. O CMV foi isolado de sangue periferico em 12,5 por cento, da urina em 23,2 por cento, da saliva em 21,9 por cento dos pacientes. Exames anatomopatologicos foram realizados em 24 pacientes, correspondendo a 60 por cento dos pacientes que evoluiram para obito durante o periodo de estudo...


Subject(s)
Adult , Middle Aged , Humans , Male , Female , Acquired Immunodeficiency Syndrome/complications , Cytomegalovirus Infections/complications , Acquired Immunodeficiency Syndrome/blood , Acquired Immunodeficiency Syndrome/diagnosis , Acquired Immunodeficiency Syndrome/urine , Cytomegalovirus Infections/blood , Cytomegalovirus Infections/diagnosis , Cytomegalovirus Infections/urine , Cytomegalovirus/isolation & purification , Saliva/microbiology
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