RESUMO
The pursuit of construction sustainability has driven the use of partially or wholly waste-based eco-materials. New applications are being sought for recycled aggregate (RA) to further the use of this material and ensure the survival of the construction and demolition waste (C&DW) industry. RA, currently used in the construction of pavements, fills and embankments and only incipiently to manufacture structural or non-structural concrete, is a mere 8.99% of the total aggregate extracted in EU countries where RA is produced. Against this backdrop, the utility of this study lies in the overview afforded of RA typology, the application of the product in bound or unbound materials, the pursuit of new applications, structural or otherwise, and the assessment of daily maximum output by C&DW recycling facilities in Spain. The findings show that irrespective of its origin, RA most commonly adopts the form of wet mix macadam, gravel or sand used primarily in unbound applications with only dubious quality standards. Plant managers contend that RA from clean waste can feasibly be used in bound applications that require higher-performance materials. Maximum daily output varies widely, with capacity under 200 t d-1 in 30% of the plants. One of the conclusions drawn is that the current business model is in need of revision, with an emphasis on aggregate quality control (certification) and the adoption of technology for separating out impurities, pollutants and undesirable materials to improve the quality of RA.
Assuntos
Indústria da Construção , Gerenciamento de Resíduos , Materiais de Construção , Resíduos Industriais , Reciclagem , EspanhaRESUMO
In the present work, we carried out a morphological, biometrical and molecular study of the species Archaeopsylla erinacei (Bouché, 1835) and their subspecies: Archaeopsylla erinacei erinacei (Bouché, 1835) and Archaeopsylla erinacei maura (Jordan & Rothschild, 1912) isolated from hedgehogs (Erinaceus europaeus) from different geographical regions (Seville and Corse). We have found morphological differences in females of A. erinacei from the same geographical origin that did not correspond with molecular differences. We suggest that some morphological characters traditionally used to discriminate females of both subspecies should be revised as well as we set the total length of the spermatheca as a valid criterion in order to discriminate between both subspecies. The Internal Transcribed Spacers 1 and 2 (ITS1, ITS2) and partial 18S rRNA gene, and partial cytochrome c-oxidase 1 (cox1) and cytochrome b (cytb) mtDNA gene sequences were determined to clarify the taxonomic status of these taxa and to assess intra-specific and intra-population similarity. In addition, a phylogenetic analysis with other species of fleas using Bayesian and Maximum Likelihood analysis was performed. All molecular markers used, except 18S, showed molecular differences between populations corresponding with geographical origins. Thus, based on the phylogenetic and molecular study of two nuclear markers (ITS1, ITS2) and two mitochondrial markers (cox1 and cytb), as well as concatenated sequences of both subspecies, we reported the existence of two geographical genetic lineages in A. erinacei corresponding with two different subspecies: A. e. erinacei (Corse, France) and A. e. maura (Seville, Spain), that could be discriminated by polymerase chain reaction-linked random-fragment-length polymorphism.
Assuntos
Sifonápteros/anatomia & histologia , Sifonápteros/genética , Animais , DNA Mitocondrial/análise , DNA Espaçador Ribossômico/análise , Feminino , França , Proteínas de Insetos/análise , Masculino , Filogenia , RNA Ribossômico 18S/análise , Análise de Sequência de DNA , Análise de Sequência de RNA , Sifonápteros/classificação , Sifonápteros/enzimologia , EspanhaRESUMO
In the present work, we carried out a comparative molecular study of Stenoponia tripectinata tripectinata isolated from Mus musculus from the Canary Islands, Spain. The Internal Transcribed Spacers 1 and 2 (ITS1, ITS2) and 18S ribosomal RNA partial gene and cytochrome c-oxidase 1 (cox1) mitochondrial DNA partial gene sequences of this subspecies were determined to clarify the taxonomic status of this subspecies and to assess inter-population variation and inter-specific sequence differences. In addition, we have carried out a comparative phylogenetic study with other species of fleas using Bayesian, Maximum Parsimony, Maximum Likelihood and Neighbor-Joining analysis. A geographical signal was detected between the cox1 partial gene sequences of S. t. tripectinata isolated from M. musculus from different islands and those isolated from Apodemus sylvaticus from the Iberian Peninsula. Our results assess the monophyletic origin of Stenoponiinae and a different genetic lineage from Ctenophthalmidae. Thus, the elevation of subfamily Stenoponiinae to family level (Stenoponiidae) is suggested.
Assuntos
Variação Genética , Filogenia , Sifonápteros/genética , Animais , DNA Intergênico/química , DNA Mitocondrial/química , Complexo IV da Cadeia de Transporte de Elétrons/química , Complexo IV da Cadeia de Transporte de Elétrons/genética , Funções Verossimilhança , Camundongos , Filogeografia , RNA Ribossômico 18S/química , Sifonápteros/anatomia & histologia , Espanha , Especificidade da EspécieRESUMO
In the present work, a comparative morphological, biometrical and molecular study of Ctenocephalides spp. isolated from dogs (Canis lupus familiaris) from different geographical regions (Spain, Iran, and South Africa) has been carried out. The internal transcribed spacer 1 (ITS1) sequences of Ctenocephalides felis and Ctenocephalides canis collected from dogs from different geographical regions have been determined to clarify the taxonomic status of these species and to assess intraspecific variation and interspecific sequence differences. In addition, a phylogenetic analysis based on ITS1 sequences has been performed. Four different morphological populations were observed in the individuals of C. felis collected from dogs from different geographical locations. Nevertheless, the comparative study of the ITS1 sequences of the different morphological populations observed in C. felis did not show molecular differences. The results showed clear molecular differences between C. felis and C. canis and some specific recognition sites for endonucleases were detected between both species. Thus, BfrBI and DraI sites have diagnostic value for specific determination in C. felis. The phylogenetic tree based on the ITS1 sequences of C. felis and C. canis revealed that all the populations of C. felis from different geographical regions clustered together and separated, with high bootstrap values, from C. canis. We conclude that ITS1 region is a useful tool to approach different taxonomic and phylogenetic questions in Ctenocephalides species.
Assuntos
Biometria , Ctenocephalides/anatomia & histologia , Ctenocephalides/genética , Doenças do Cão/parasitologia , Variação Genética , Filogeografia , Animais , Análise por Conglomerados , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Cães , Ectoparasitoses/parasitologia , Ectoparasitoses/veterinária , Irã (Geográfico) , Dados de Sequência Molecular , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , África do Sul , EspanhaRESUMO
BACKGROUND: Electronic health records (EHRs) are used to register important health-related information, such as allergic conditions, and contribute to the safety and quality of medical care. OBJECTIVES: To evaluate the use of allergy alert entries in EHRs and to establish the allergy profile of hospitalized patients. METHODS: Allergy data recorded in EHRs were analyzed in a cross-sectional, observational, descriptive study of patients admitted to the hospital from January 1 through June 30, 2011. RESULTS: A total of 15,534 patients were admitted to the hospital during the study period. The rate of inclusion of allergy information in the EHRs was 64.4%. In 2,106 patients an alert was activated to declare an allergy, intolerance, or any other type of adverse reaction. Drugs were the most common responsible agent (74.4%), followed by foods (12.6%) and materials (4.8%). Entries for drug allergy or intolerance were more common in females (64.8%) than males, with a significant statistical difference (P < .01), and increased proportionally with age. Entries for food allergy or intolerance were also more common in females (58.0%) than males (P < .01), but this trend was reversed in the 0- to 15-year-old age group. By contrast, the entries for food allergy or intolerance decreased proportionally with age. In 7,907 cases the EHRs revealed that patients were free of allergies, intolerances, or any other type of adverse reactions. CONCLUSION: Drug allergy was the most frequently reported allergic condition, followed by foods and materials. Allergy alerts vary depending on age and sex. The proper use of a system for allergy alerts included in EHRs provides valuable information about hospitalized patients, contributing to the improvement of clinical practice.
Assuntos
Fatores Etários , Registros Eletrônicos de Saúde/estatística & dados numéricos , Hipersensibilidade/epidemiologia , Sistemas de Registro de Ordens Médicas/estatística & dados numéricos , Fatores Sexuais , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Alérgenos/efeitos adversos , Alérgenos/imunologia , Criança , Pré-Escolar , Estudos Transversais , Feminino , Hospitalização , Humanos , Hipersensibilidade/fisiopatologia , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Prevalência , Adulto JovemRESUMO
OBJECTIVE: To characterize patterns of chronic ocular disease in patients with Stevens-Johnson syndrome (SJS) and its variant toxic epidermal necrolysis (TEN), and to describe their response to treatment. METHODS: Retrospective case series. A review of hospital records of 30 patients (60 eyes) with ocular manifestations of SJS or TEN was carried out. The principal outcome measure was to identify and classify the patterns of chronic ocular disease in SJS and TEN. The secondary outcome measure was the response to treatment. RESULTS: Patterns of chronic ocular disease observed after the acute episode included: mild/moderate SJS, severe SJS, ocular surface failure (SJS-OSF), recurrent episodic inflammation (SJS-RI), scleritis (SJS-S) and progressive conjunctival cicatrisation resembling mucous membrane pemphigoid (SJS-MMP). The median follow-up was 5 years (range 0-29). 19 patients (29 eyes (48%)) developed SJS-OSF, SJS-RI, SJS-S or SJS-MMP during follow-up. SJS-OSF was present in 12 patients (18 eyes (30%)). In 5 patients (eight eyes) this developed 1 year after the acute illness, without any further inflammatory episodes; it was associated with SJS-RI in 1 patient (2 eyes), with SJS-RI and SJS-S in 1 patient (1 eye), with SJS-S in 1 patient (1 eye) and with SJS-MMP in 4 patients (6 eyes). Episodes of SJS-RI occurred in 4 patients (7 eyes (12%)). The median time from acute disease to the first episode of SJS-RI was 8.5 years (range 5-63). SJS-S developed in 2 patients (4 eyes (7%)), of which 2 eyes subsequently developed SJS-OSF. SJS-MMP developed in 5 patients (10 eyes (16.6%)). The median duration from the acute stage to the diagnosis of SJS-MMP was 2 years (range 1-14). Immunosuppressive therapy successfully controlled inflammation in 10/10 patients with SJS-MMP, SJS-RI or SJS-S. CONCLUSION: Ocular disease in SJS/TEN is not limited solely to the sequelae of the acute phase illness. Patients and physicians need to know that ocular disease progression, due to surface failure and/or acute inflammatory conditions, may occur at variable periods following the acute disease episode. Recognition of this, and prompt access to specialist services, may optimise management of these uncommon patterns of disease in SJS.
Assuntos
Anti-Inflamatórios/uso terapêutico , Oftalmopatias/etiologia , Síndrome de Stevens-Johnson/complicações , Adolescente , Adulto , Idoso , Criança , Doença Crônica , Oftalmopatias/tratamento farmacológico , Oftalmopatias/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Síndrome de Stevens-Johnson/tratamento farmacológico , Síndrome de Stevens-Johnson/patologiaRESUMO
At the present work, we carried out a morph-biometrical and molecular study of Trichuris species isolated from Camelus dromedarius from Iran and from Ovis aries from South Africa comparatively with other species of Trichuris from different herbivorous hosts and geographical regions. The population from camels from Iran was identified as Trichuris globulosa. Two different morphometrically populations of Trichuris sp. from sheep from South Africa were identified: Trichuris ovis and Trichuris skrjabini. Ribosomal data did not reveal significate differences in the ITS2 sequences between T. ovis and T. globulosa to assess a specific determination. The mitochondrial data suggest that T. globulosa constitute a different genetic lineage to T. ovis. Cytochrome c-oxidase and cytochrome b partial gene sequences corroborated the existence of a different genetic lineage of T. ovis from sheep of South Africa that would be closely related to the populations of T. globulosa from camels from Iran. The cytochrome c-oxidase and cytochrome b partial gene sequences of T. globulosa have been reported for the first time.
Assuntos
Doenças dos Ovinos/parasitologia , Tricuríase/veterinária , Trichuris/classificação , Animais , Camelus/parasitologia , Citocromos b/genética , DNA de Helmintos/genética , DNA Mitocondrial/genética , DNA Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Feminino , Genes de Helmintos , Herbivoria , Masculino , Filogenia , Análise de Sequência de DNA , Ovinos , Especificidade da Espécie , Tricuríase/parasitologia , Trichuris/anatomia & histologia , Trichuris/genéticaRESUMO
Levels in serum of components of complement were studied in a group of 10 patients with mixed cryoglobulinemia. The profiles found in most patients showed decreased levels of the early complement components C1, C4, and C2, with normal levels of C3. Experiments performed to define the mechanism(s) responsible for this unusual complement profile showed that activation of the early complement components in serum was due to the activation of the classical pathway by mixed cryoglobulins. They also showed that the characteristic lack of effect on C3 was due to the action of a previously unrecognized regulatory mechanism upon C3 convertase of the classical pathway mediated by 2 normal serum proteins, namely, the C4 binding protein (C4-bp) and the C3b inactivator (C3bINA).
Assuntos
Ativação do Complemento , Proteínas do Sistema Complemento , Crioglobulinas , Enzimas Ativadoras do Complemento/metabolismo , Complemento C4 , HumanosRESUMO
The over-all synthesis of DNA, RNA and protein was studied during the asexual cell cycle of Plasmodium falciparum. A method for stringent synchronization of the culture was developed. Rates of synthesis of the three macromolecules were determined every four hours by labelling the parasites with radioactive precursors during 30 min pulses. There was a peak of synthesis of DNA at 44 hours. Between 33 and 45 hours the rate of synthesis of DNA was exponential. Maximum RNA synthesis was reached at 36 hours. For proteins there were two peaks, one at 24 and the other at 40 hours. Partial degradation of the RNA synthesized by schizonts was detected.
Assuntos
DNA/biossíntese , Plasmodium falciparum/metabolismo , Biossíntese de Proteínas , RNA/biossíntese , Ciclo Celular , Plasmodium falciparum/citologia , Fatores de TempoRESUMO
Common variable immunodeficiency (CVID) is a heterogeneous immunodeficiency syndrome characterized by hypogammaglobulinemia, recurrent bacterial infections, and various immunologic abnormalities. The clinical presentation is generally that of recurrent pyogenic sinopulmonary infections. Our objectives were to study the prevalence of lung involvement and the response to intravenous immunoglobulin replacement therapy in 19 patients with CVID. Nineteen patients (12 men) with a mean age (SD) of 33.1 (17.1) years had a previous diagnosis of CVID and were treated with intravenous immunoglobulin replacement. All patients underwent complete pulmonary function tests and high-resolution computed tomography (HRCT) examination. Bronchiectasis was diagnosed in 11 (58%) patients and eight (42%) were multi-lobar bronchiectasis. Chronic airflow limitation (CAL) was present in 10 (53%) patients and a restrictive pattern was seen in one case. Eleven patients (58%) presented a decrease in single-breath carbon monoxide diffusing capacity of the lung (DL(CO)). Before intravenous immunoglobulin replacement therapy (INIRT), 84% of patients had suffered from at least one episode of pneumonia. Episodes of lower respiratory tract infection decreased significantly from 0.28 per patient and year before replacement therapy to 0.16 per patient and year after treatment. The mean duration of replacement therapy was 7.5 years. In conclusion lung involvement was frequent in patients with CVID. Long-term administration of intravenous gammaglobulin resulted in a substantial reduction of pneumonic episodes.
Assuntos
Imunodeficiência de Variável Comum/complicações , Doenças Respiratórias/etiologia , Adolescente , Adulto , Idoso , Distribuição de Qui-Quadrado , Criança , Imunodeficiência de Variável Comum/diagnóstico , Imunodeficiência de Variável Comum/terapia , Feminino , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Testes de Função Respiratória , Doenças Respiratórias/diagnóstico , Doenças Respiratórias/terapia , Estatísticas não Paramétricas , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
Adults of Trichuris skrjahini have been isolated from the cecum of caprine hosts (Capra hircus), Trichuris ovis and Trichuris globulosa from Ovis aries (sheep) and C. hircus (goats), and Trichuris leporis from Lepus europaeus (rabbits) in Spain. Genomic DNA was isolated and the ITS1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced by polymerase chain reaction (PCR) techniques. The ITS1 of T. skrjabini, T. ovis, T. globulosa, and T. leporis was 495, 757, 757, and 536 nucleotides in length, respectively, and had G + C contents of 59.6, 58.7, 58.7, and 60.8%, respectively. Intraindividual variation was detected in the ITSI sequences of the 4 species. Furthermore, the 5.8S sequences of T. skrjabini, T. ovis, T. globulosa, and T. leporis were compared. A total of 157, 152, 153, and 157 nucleotides in length was observed in the 5.8S sequences of these 4 species, respectively. There were no sequence differences of ITS1 and 5.8S products between T. ovis and T. globulosa. Nevertheless, clear differences were detected between the ITS1 sequences of T. skrjabini, T. ovis, T. leporis, Trichuris muris, and T. arvicolae. The ITS2 fragment from the rDNA of T. skrjabini was sequenced. A comparative study of the ITS2 sequence of T. skrjabini with the previously published ITS2 sequence data of T. ovis, T. leporis, T. muris, and T. arvicolae suggested that the combined use of sequence data from both spacers would be useful in the molecular characterization of trichurid parasites.
Assuntos
DNA Espaçador Ribossômico/química , Doenças das Cabras/parasitologia , Coelhos/parasitologia , Doenças dos Ovinos/parasitologia , Tricuríase/veterinária , Trichuris/genética , Animais , Sequência de Bases , Ceco/parasitologia , Sequência Consenso , DNA de Helmintos/química , Cabras , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 5,8S/genética , Ovinos , Espanha , Tricuríase/parasitologia , Trichuris/classificaçãoRESUMO
Perioperative management of angiotensin-converting enzyme inhibitors (ACEI) is controversial because of associated hypertensive episodes during induction and maintenance of anesthesia. A 71-year-old woman with a non-functioning thyroid node was scheduled for thyroid lobectomy. Her medical history included high blood pressure and she was being chronically treated with ACEI, which were taken until the morning of surgery. After induction of anesthesia, arterial hypotension refractory to crystalloid therapy developed and worsened in spite of administration of a gelatin-type colloid (Gelafundina). The patient did not respond to ephedrine or dopamine and required stabilization with adrenalin in continuous perfusion for 12 hours. Later evolution was satisfactory and recovery took place without sequelae. We discuss the anesthetic implications of chronic ACEI treatment and possible hemodynamic repercussions of associated administration with gelatin-type solutions or human albumin.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/efeitos adversos , Cilazapril/efeitos adversos , Hipotensão/induzido quimicamente , Complicações Intraoperatórias/induzido quimicamente , Complicações Pós-Operatórias/induzido quimicamente , Idoso , Inibidores da Enzima Conversora de Angiotensina/administração & dosagem , Cilazapril/administração & dosagem , Coloides/uso terapêutico , Soluções Cristaloides , Dopamina/uso terapêutico , Resistência a Medicamentos , Efedrina/uso terapêutico , Epinefrina/administração & dosagem , Epinefrina/uso terapêutico , Feminino , Gelatina/uso terapêutico , Humanos , Hipertensão/complicações , Hipertensão/tratamento farmacológico , Hipotensão/tratamento farmacológico , Infusões Parenterais , Complicações Intraoperatórias/tratamento farmacológico , Soluções Isotônicas , Substitutos do Plasma/uso terapêutico , Complicações Pós-Operatórias/tratamento farmacológico , Nódulo da Glândula Tireoide/complicações , Nódulo da Glândula Tireoide/cirurgia , TireoidectomiaRESUMO
Comparative morphological, biometrical and molecular studies of Trichuris discolor isolated from Bos taurus from Spain and Iran was carried out. Furthermore, Trichuris ovis isolated from B. taurus and Capra hircus from Spain has been, molecularly, analyzed. Morphological studies revealed clear differences between T. ovis and T. discolor isolated from B. taurus but differences were not observed between populations of T. discolor isolated from different geographical regions. Nevertheless, the molecular studies based on the amplification and sequencing of the internal transcribed spacers 1 and 2 ribosomal DNA and 16S partial gene mitochondrial DNA showed clear differences between both populations of T. discolor from Spain and Iran suggesting two cryptic species. Phylogenetic studies corroborated these data. Thus, phylogenetic trees based on ITS1, ITS2 and 16S partial gene sequences showed that individuals of T. discolor from B. taurus from Iran clustered together and separated, with high bootstrap values, of T. discolor isolated from B. taurus from Spain, while populations of T. ovis from B. taurus and C. hircus from Spain clustered together but separated with high bootstrap values of both populations of T. discolor. Furthermore, a comparative phylogenetic study has been carried out with the ITS1and ITS2 sequences of Trichuris species from different hosts. Three clades were observed: the first clustered all the species of Trichuris parasitizing herbivores (T. discolor, T. ovis, Trichuris leporis and Trichuris skrjabini), the second clustered all the species of Trichuris parasitizing omnivores (Trichuris trichiura and Trichuris suis) and finally, the third clustered species of Trichuris parasitizing carnivores (Trichuris muris, Trichuris arvicolae and Trichuris vulpis).
Assuntos
Doenças dos Bovinos/parasitologia , DNA Mitocondrial/genética , DNA Espaçador Ribossômico/genética , RNA Ribossômico 16S/genética , Tricuríase/veterinária , Trichuris/genética , Animais , Sequência de Bases , Bovinos , Doenças dos Bovinos/epidemiologia , Demografia , Feminino , Marcadores Genéticos , Irã (Geográfico)/epidemiologia , Masculino , Dados de Sequência Molecular , Espanha/epidemiologia , Tricuríase/epidemiologia , Tricuríase/parasitologia , Trichuris/fisiologiaRESUMO
The partial mitochondrial cytochrome c-oxidase subunit 1 gene (cox 1) and partial mitochondrial 16S ribosomal DNA of Trichuris skrjabini (Baskakov 1924) isolated from Capra hircus have been amplified and sequenced. The analyses of multiple sequence alignments of mitochondrial 16S rDNA and cox 1 of T. skrjabini revealed high homology with those of Trichinella species. For the first time, the mitochondrial DNA gene sequences of one species of trichurid nematode have been cited.
Assuntos
DNA de Helmintos/genética , DNA Mitocondrial/genética , DNA Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Trichuris/genética , Animais , DNA de Helmintos/química , DNA Mitocondrial/química , DNA Ribossômico/química , Cabras/parasitologia , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência , Trichinella/genética , Trichuris/enzimologia , Trichuris/isolamento & purificaçãoRESUMO
In this paper, a morphological and biometrical study by optical microscopy and scanning electronic microscopy (SEM) of Trichuris suis isolated from different hosts (Sus scrofa domestica and Sus scrofa scrofa) and Trichuris trichiura isolated from chimpanzee, has been carried out. Our results demonstrate the existence of typical pericloacal papillae in both species. Biometrical parameters of T. suis and T. trichiura overlapped but males and females of T. trichiura tended to be shorter and thinner than those of T. suis. Our results suggest that T. suis and T. trichiura cannot be differentiated using standard procedures as morphological and biometrical determinations. Thus, the ITS1-5.8S-ITS2 region of the ribosomal DNA was sequenced to allow a differentiation between T. suis and T. trichiura on genetic level. The ITS1 and ITS2 sequences derived from T. trichiura eggs isolated from feces of primates (Colobus guereza kikuyensis and Nomascus gabriellae) showed clear differences to the respective sequences of T. suis derived from eggs of different porcine hosts. The 5.8S gene was similar between the two species. Sequences obtained from different populations of the same species showed no significant differences indicating that the ITS1-5.8S-ITS2 sequences reported in this study are representative for T. trichiura and T. suis, respectively. Phylogenetic relationships have been determined attending to the ITS1 and ITS2 sequences from different species of the genus Trichuris. In conclusion, T. trichiura and T. suis are considered to be closely related but genetically different species. Both species can be easily and reliably distinguished by a PCR-RFLP analysis of the ITS1 and ITS2 sequences with different restriction enzymes.
Assuntos
Tricuríase/veterinária , Trichuris/anatomia & histologia , Trichuris/classificação , Animais , Sequência de Bases , Análise por Conglomerados , DNA de Helmintos/química , DNA de Helmintos/genética , DNA Intergênico/química , DNA Intergênico/genética , Feminino , Masculino , Microscopia/métodos , Microscopia Eletrônica de Varredura/métodos , Dados de Sequência Molecular , Pan troglodytes/parasitologia , Filogenia , RNA Ribossômico 5,8S/química , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Homologia de Sequência , Sus scrofa/parasitologia , Tricuríase/parasitologia , Trichuris/genética , Trichuris/isolamento & purificaçãoRESUMO
Adult worms of Trichuris ovis and T. globulosa were collected from Ovis aries (sheep) and Capra hircus (goats). T. suis was isolated from Sus scrofa domestica (swine) and T. leporis was isolated from Lepus europaeus (rabbits) in Spain. Genomic DNA was isolated and a ribosomal internal transcribed spacer (ITS2) was amplified and sequenced using polymerase-chain-reaction (PCR) techniques. The ITS2 of T. ovis and T. globulosa was 407 nucleotides in length and had a GC content of about 62%. Furthermore, the ITS2 of T. suis and T. leporis was 534 and 418 nucleotides in length and had a GC content of about 64.8% and 62.4%, respectively. There was evidence of slight variation in the sequence within individuals of all species analyzed, indicating intraindividual variation in the sequence of different copies of the ribosomal DNA. Furthermore, low-level intraspecific variation was detected. Sequence analyses of ITS2 products of T. ovis and T. globulosa demonstrated no sequence difference between them. Nevertheless, differences were detected between the ITS2 sequences of T. suis, T. leporis, and T. ovis, indicating that Trichuris species can reliably be differentiated by their ITS2 sequences and PCR-linked restriction-fragment-length polymorphism (RFLP).
Assuntos
DNA de Helmintos/genética , DNA Espaçador Ribossômico/genética , Trichuris/classificação , Trichuris/genética , Animais , Composição de Bases , Sequência de Bases , Enzimas de Restrição do DNA , DNA de Helmintos/química , DNA Espaçador Ribossômico/química , Genes de RNAr , Variação Genética , Cabras/parasitologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Coelhos/parasitologia , Mapeamento por Restrição , Ovinos/parasitologia , Especificidade da Espécie , Suínos/parasitologiaRESUMO
Impression cytology is a simple and painless procedure that allows the collection of the superficial layers of the conjunctival epithelium. Each sample is assigned a grade of epithelial metaplasia, and goblet cell density is calculated in each one. We have studied the superior and temporal bulbar conjunctiva of dry eyes and have compared it with that of normal controls. In normal and dry eyes we find a statistically significant difference both in goblet cell density and grade of metaplasia, between superior and temporal bulbar conjunctiva. The differences in grade of metaplasia and goblet cell density between normal and dry eyes are significant in the superior conjunctiva, but in the temporal conjunctiva we only find significant differences in grade of metaplasia.
Assuntos
Técnicas Citológicas , Ceratoconjuntivite Seca/patologia , Idoso , Túnica Conjuntiva/patologia , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Trichuris muris has been isolated from murid hosts ( Apodemus sylvaticus and Mus musculus) and Trichuris arvicolae from arvicolid rodents in Barcelona, Spain. Genomic DNA was isolated and the ITS1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced using polymerase chain reaction techniques. The ITS2 of both populations isolated from Apodemus and Mus was 382 nucleotides in length and had a GC content of about 60.73%, while the ITS2 of T. arvicolae was 442 nucleotides in length and had a GC content of about 59.8%. Furthermore, the ITS1 of Trichuris from murids was 448 nucleotides in length and had a GC content of about 56.47%, while T. arvicolae was 446 nucleotides in length and had 57.62% of GC content. A total of 161 and 173 nucleotides were observed along the 5.8S gene of T. murisand T. arvicolae, respectively; This difference in nucleotides was due to the insertion of a DNA segment (transposon) in the 5.8S sequence of the latter species. Slight intraindividual and intraspecific variations were detected in the rDNA of both species. The presence of microsatellites was observed in all of the individuals assayed. Sequence analysis of the internal transcribed spacers and the 5.8S gene demonstrated no sequence differences between T. muris isolated from both of its murid hosts. Nevertheless, clear differences were detected between the ITS2, ITS1 and 5.8S gene of T. muris and T. arvicolae. This corroborates the existence of two separate Trichuris species in murid and arvicolid hosts. Furthermore, a phylogenetic analysis was carried out and endonucleases restriction maps were elaborated for both species.
Assuntos
Arvicolinae/parasitologia , DNA Espaçador Ribossômico/análise , Muridae/parasitologia , RNA Ribossômico 5,8S/genética , Trichuris/isolamento & purificação , Animais , Sequência de Bases , DNA de Helmintos/análise , Camundongos , Modelos Genéticos , Dados de Sequência Molecular , Técnicas de Amplificação de Ácido Nucleico , Filogenia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Trichuris/genéticaRESUMO
PURPOSE: To investigate the presence of three C-C chemokines (monocyte chemotactic protein-1 [MCP-1], macrophage inflammatory protein-1alpha [MIP-1alpha], and MIP-1beta) in vitreous samples from eyes with proliferative vitreoretinopathy (PVR), proliferative diabetic retinopathy (PDR), or retinal detachment (RD). SUBJECTS AND METHODS: Vitreous samples were obtained and assayed by solid phase enzyme-linked immunosorbent assay for the chemokines MCP-1, MIP-1alpha, and MIP-1beta in a prospective study of 43 consecutive patients. Eighteen samples from cadaveric control eyes were also assayed. RESULTS: Monocyte chemotactic protein-1 was detected in all samples. The vitreous of eyes with vitreoretinal disorders showed significantly higher levels than the vitreous of cadaveric control eyes (P < or = 0.05). Median level (5th-95th percentile) in the PVR cases (n = 20) was 890.18 pg/mL (286.04-1806.20); in RD (n = 8), 296.69 pg/mL (171.44-1310.02); and in PDR (n = 15), 434.60 pg/mL (124.56-1092.94). In the cadaveric control eyes (n = 18), median level was 83.97 pg/mL (26.09-208.38). Macrophage inflammatory protein-1alpha and MIP-1beta were not detected in any samples. CONCLUSION: Monocyte chemotactic protein-1 might be involved in the recruitment of macrophages and monocytes into the vitreous of eyes with proliferative vitreoretinal disorders.
Assuntos
Quimiocinas CC/metabolismo , Retinopatia Diabética/metabolismo , Vitreorretinopatia Proliferativa/metabolismo , Corpo Vítreo/metabolismo , Biomarcadores , Cadáver , Quimiocina CCL2/metabolismo , Quimiocina CCL3 , Quimiocina CCL4 , Retinopatia Diabética/cirurgia , Ensaio de Imunoadsorção Enzimática , Seguimentos , Humanos , Proteínas Inflamatórias de Macrófagos/metabolismo , Descolamento Retiniano/metabolismo , Descolamento Retiniano/cirurgia , Vitrectomia , Vitreorretinopatia Proliferativa/cirurgia , Corpo Vítreo/cirurgiaRESUMO
The complete internal transcribed spacer 1 (ITS1), 5.8S rDNA and ITS2 region of the ribosomal DNA from 11 species of rhinonyssid mites ( Tinaminyssus columbae, T. minisetosum, T. sartbaevi, T. bubulci, T. melloi, T. streptopelioides, Sternostoma fulicae, S. boydi, S. strandtmanni, S. turdi, Rhinonyssus tringae) were sequenced to assess the utility of this genomic region in resolving taxonomic questions in this group and to estimate phylogenetic relationships between species. Two different geographic locations of T. melloi and T. streptopelioides were analyzed to detect intraspecies variation. Our study shows that ribosomal sequences can help to discriminate between T. melloi and T. sartbaevi, which are morphologically very close and difficult to separate by classic methods. The resulting phylogenetic tree shows some differences from the current taxonomy of the family Rhinonyssidae. This study appeals for the revision of the taxonomic status of S. boydi and closely related species which parasitize aquatic birds and suggests the synonymy of S. boydi and S. strandtmanni, despite the different hosts of the two mites.