RESUMO
INTRODUCTION: As large scale metabolic phenotyping is increasingly employed in preclinical studies and in the investigation of human health and disease the current LC-MS/MS profiling methodologies adopted for large sample sets can result in lengthy analysis times, putting strain on available resources. As a result of these pressures rapid methods of untargeted analysis may have value where large numbers of samples require screening. OBJECTIVES: To develop, characterise and evaluate a rapid UHP-HILIC-MS-based method for the analysis of polar metabolites in rat urine and then extend the capabilities of this approach by the addition of IMS to the system. METHODS: A rapid untargeted HILIC LC-MS/MS profiling method for the analysis of small polar molecules has been developed. The 3.3 min separation used a Waters BEH amide (1 mm ID) analytical column on a Waters Synapt G2-Si Q-Tof enabled with ion mobility spectrometry (IMS). The methodology, was applied to the metabolic profiling of a series of rodent urine samples from vehicle-treated control rats and animals administered tienilic acid. The same separation was subsequently linked to IMS and MS to evaluate the benefits that IMS might provide for metabolome characterisation. RESULTS: The rapid HILIC-MS method was successfully applied to rapid analysis of rat urine and found, based on the data generated from the data acquired for the pooled quality control samples analysed at regular intervals throughout the analysis, to be robust. Peak area and retention times for the compounds detected in these samples showed good reproducibility across the batch. When used to profile the urine samples obtained from vehicle-dosed control and those administered tienilic acid the HILIC-MS method detected 3007 mass/retention time features. Analysis of the same samples using HILIC-IMS-MS enabled the detection of 6711 features. Provisional metabolite identification for a number of compounds was performed using the high collision energy MS/MS information compared against the Metlin MS/MS database and, in addition, both calculated and measured CCS values from an experimentally derived CCS database. CONCLUSION: A rapid metabolic profiling method for the analysis of polar metabolites has been developed. The method has the advantages of speed and both reducing sample and solvent consumption compared to conventional profiling methods. The addition of IMS added an additional dimension for feature detection and the identification of metabolites.
Assuntos
Ensaios de Triagem em Larga Escala/métodos , Metabolômica/métodos , Urina/química , Animais , Líquidos Corporais , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida/métodos , Humanos , Masculino , Metaboloma , Controle de Qualidade , Ratos/urina , Ratos Sprague-Dawley/urina , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/métodosRESUMO
Aditoprim (ADP) is a newly developed antibacterial agent in veterinary medicine. The metabolism and disposition of ADP in swine, broilers, carp and rats were investigated by using a radio tracer method combined with a radioactivity detector and a liquid chromatography/ion trap/time-of-flight mass spectrometry. After a single oral administration, more than 94% of the dose was recovered within 14 d in the four species. The urine excretion was dominant in swine and rats, making up 78% of the dose. N-monodesmethyl-ADP, N-didesmethyl-ADP, and 10 new metabolites were characterized. These metabolites were biotransformed from the process of demethylation, α-hydroxylation, N-oxidation, and NH2-glucuronidation. After an oral dose for 7 d, ADP-derived radioactivity was widely distributed in tissues, and high concentrations were especially observed in bile, liver, kidney, lung, and spleen. The radioactivity in the liver was eliminated much more slowly than in other tissues, with a half-life of 4.26, 3.38, 6.69, and 5.21 d in swine, broilers, carp, and rats, respectively. ADP, N-monodesmethyl-ADP, and N-didesmethyl-ADP were the major metabolites in edible tissues. Notably, ADP was detected with the highest concentration and the longest duration in these tissues. These findings indicated that ADP is the marker residue and the liver is the residue target tissue.
Assuntos
Difosfato de Adenosina/metabolismo , Fígado/química , Trimetoprima/análogos & derivados , Administração Oral , Animais , Carpas/urina , Galinhas/urina , Cromatografia Líquida , Espectrometria de Massas , Ratos/urina , Suínos/urina , Distribuição Tecidual , Trimetoprima/administração & dosagem , Trimetoprima/farmacocinética , Trimetoprima/urinaRESUMO
In addition to having selective potency against the molecular target(s), a compound must be able to reach its intended site of action in vivo in sufficient quantity and for the appropriate duration of time to exert a biological effect. The fate of a compound after in vivo administration depends upon its absorption, distribution, metabolism, and excretion (ADME), as well as its target residence time. The concentration of the compound in the blood, plasma, and other tissues represents the sum of all of these processes. Described in this unit are protocols for administering a compound by various routes to rats and for collecting the appropriate samples to determine the pharmacokinetics profile. The basic terms used in pharmacokinetics studies are defined, and representative examples are given to illustrate important variables to consider.
Assuntos
Farmacocinética , Anestesia por Inalação , Anestésicos Inalatórios , Animais , Coleta de Amostras Sanguíneas , Fezes/química , Feminino , Isoflurano , Masculino , Preparações Farmacêuticas/administração & dosagem , Preparações Farmacêuticas/análise , Preparações Farmacêuticas/sangue , Preparações Farmacêuticas/urina , Radioisótopos/farmacocinética , Ratos/sangue , Ratos/metabolismo , Ratos/urina , Distribuição TecidualRESUMO
The recent high-profile death of a British Olympic rower from leptospirosis has raised awareness to this uncommon but potentially fatal disease. The re-emergence of the disease abroad is well documented in the literature, but less is known about cases in the UK. The increase in participation in water sports, foreign travel and often a combination of the two, has increased the exposure of tourists subsequently returning to the UK from areas of high prevalence. Leptospirosis is a zoonotic infection. The bacteria are shed in the urine of animals to the environment from where humans are infected by incidental hosts. There is a wide spectrum of severity of symptoms, from a self-limiting febrile illness to fatal pulmonary haemorrhage, renal or liver failure. It is thought that cases remain unrecognized every year in the UK, largely due to the mild nature of symptoms and the wide differential for febrile illness and partly due to lack of awareness among clinicians. This review examines the epidemiology of leptospirosis in the UK, over the period 2006-10, the clinical features, diagnostic techniques and treatment.
Assuntos
Leptospirose/diagnóstico , Leptospirose/epidemiologia , Doença de Weil/diagnóstico , Doença de Weil/epidemiologia , Adolescente , Adulto , Idoso , Animais , Criança , Feminino , Humanos , Leptospirose/transmissão , Masculino , Pessoa de Meia-Idade , Ratos/urina , Ratos/virologia , Reino Unido/epidemiologia , Doença de Weil/transmissão , Adulto Jovem , ZoonosesRESUMO
Urine of rats and mice is the main source of allergenic proteins that can enter the respiratory tract of laboratory animal care workers. Little is known about the levels and determinants of these exposures in the United States. We investigated the relationship between activities in animal facilities and levels of personal exposure to allergen by collecting personal breathing zone dust samples from 7 caretakers during full workdays for 1 wk. Mice and rat urinary allergens in inhalable dust were quantified via immunoassay. The activities of the sampled workers were observed, and the methods of preventing exposure to allergens were recorded. Mouse urinary allergen was detected in 20 of 39 measurements, yielding a geometric mean of 0.8 ng/m(3) with a maximum of 24 ng/m(3). Washing and cleaning cages and the number of mice handled daily were the most important determinants of personal exposure to mouse urinary allergen, as identified by using multiple linear regressions that explained 51% of total variance. Personal exposures to mouse urinary allergen were associated with day-to-day variation of tasks rather than characteristics of workers. Where potential for personal exposure is the highest, protective measures (N95 masks and cage dumping stations) appeared to be used, as is appropriate. Rat urinary allergen was detected in 4 of 39 measurements; detectable concentrations were between 0.8 and 39 ng/m(3). Only persons who handled rats were exposed to rat urinary allergen. The current findings are valuable for establishing exposure levels against which comparisons of improvement or deterioration of personal exposures can be made.
Assuntos
Alérgenos/efeitos adversos , Criação de Animais Domésticos , Animais de Laboratório/imunologia , Poeira/imunologia , Exposição Ocupacional , Aerossóis , Alérgenos/imunologia , Alérgenos/urina , Animais , Animais de Laboratório/urina , Gatos , Feminino , Humanos , Laboratórios , Masculino , Máscaras , Camundongos/imunologia , Camundongos/urina , Exposição Ocupacional/prevenção & controle , Material Particulado/efeitos adversos , Material Particulado/análise , Roupa de Proteção , Coelhos , Ratos/imunologia , Ratos/urina , Suínos , Fatores de TempoRESUMO
BACKGROUND: Sensitization to rats and mice can develop in laboratory animal workers exposed to only one species. Reasons for this dual sensitization are unclear but may reflect a genetic predisposition to developing allergy (atopy) or alternatively cross-reactivity between rat and mouse urinary allergens. We examined cross-reactivity between rat and mouse urine and the effect atopy has on dual sensitization in laboratory animal workers. METHODS: In a cross-sectional study the frequency of sensitization to rat and/or mouse was analysed in 498 employees exposed to both rat and mouse at work and 220 to rat only. RAST inhibitions, western blots and blot inhibitions were carried out on a subset of five individuals to assess cross-reactivity. RESULTS: Fourteen per cent of workers were sensitized to rats and 9% to mouse. Over half (62%) of rat sensitized individuals were also mouse sensitized and the majority (91%) of mouse sensitized individuals were also rat sensitized. IgE cross-reactivity was demonstrated between rat and mouse urine using RAST inhibitions. Rates of atopy did not differ between rat only sensitized individuals compared with those sensitized to both species. Sensitization to cats and rabbits was more common amongst those with dual sensitization. CONCLUSIONS: Dual sensitization to rat and mouse reflects IgE cross-reactivity rather than atopy. Individuals with dual sensitization are more likely to be sensitized to other animal allergens. These findings will have implications for individuals working with only one rodent species who develop sensitization and symptoms to be aware of the potential for allergy to other species.
Assuntos
Alérgenos/urina , Animais de Laboratório/imunologia , Hipersensibilidade/etiologia , Camundongos/imunologia , Exposição Ocupacional , Ratos/imunologia , Adolescente , Adulto , Idoso , Alérgenos/química , Animais , Western Blotting , Reações Cruzadas , Estudos Transversais , Feminino , Humanos , Imunoglobulina E/sangue , Masculino , Camundongos/urina , Pessoa de Meia-Idade , Modelos Moleculares , Ratos/urina , Testes CutâneosRESUMO
In conventional pharmacological studies, intersubject differences within an animal strain are normally neglected, leading to variations in pharmacological outcomes in response to the same stimulus. Using two classical experimental models, the Streptozotocin (STZ)-induced diabetic model of Wistar rats and the high-energy, diet-induced obesity model of Sprague-Dawley rats, we demonstrate that the different outcomes of STZ or diet intervention are closely associated with variation in predose (baseline) urinary metabolic profiles of the rats. The pharmacometabonomic analysis of predose metabolic profiles indicates that the intersubject difference is, to a great extent, associated with gut-microbiota, which predisposes different pathophysiological outcomes upon diet alteration or chemical stimulus. We hypothesize that there may exist an important association between observations from these two models and the obese/diabetic human population in that subtle variations in metabolic phenotype may predetermine different systems' responses to xenobiotic perturbation, ultimately leading to varied pathophysiological processes. Results from two independent models also suggest that the pharmacometabonomics approach is of great importance in the study of pharmacology and clinical drug evaluations, where endogenous metabolite signatures of predose individuals should be taken into consideration to minimize intersubject difference and the resulting variation in the postdose pharmacological outcomes.
Assuntos
Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Intestinos/microbiologia , Ratos/urina , Xenobióticos/farmacologia , Animais , Glicemia/análise , Peso Corporal , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/urina , Dieta , Metabolismo Energético , Obesidade/metabolismo , Obesidade/urina , Ratos/microbiologia , Ratos Sprague-Dawley , Ratos WistarRESUMO
Rats are used routinely for the discovery of new pharmaceuticals and for toxicology testing to fulfill regulatory requirements. In 1999, a survey showed that 80% of all rodents housed in toxicology studies were housed in wire-bottom cages. However, both the National Research Council and Association for the Assessment and Accreditation of Laboratory Animal Care, International, recommend housing rats in solid-bottom cages with bedding. In this study 2 groups of male Sprague Dawley rats were housed in the same room for 4 wk and provided the same food and water by the same husbandry staff person. The only variable in the study was the type of housing. One group was housed in solid-bottom polycarbonate cages with bedding and the other group in standard wire-bottom caging. Clinical pathology laboratory evaluations of complete blood count, serum chemistries, urinalysis, urine creatinine, urine corticosterone, blood coagulation, and hepatic cytochrome P450 isoenzyme mRNA levels were performed. No clinically relevant differences were found between the 2 groups for any of the laboratory data.
Assuntos
Abrigo para Animais , Ratos/metabolismo , Testes de Toxicidade , Bem-Estar do Animal , Animais , Contagem de Células Sanguíneas/veterinária , Testes de Coagulação Sanguínea/veterinária , Peso Corporal , Corticosterona/urina , Creatinina/urina , Sistema Enzimático do Citocromo P-450/metabolismo , Testes Hematológicos/veterinária , Abrigo para Animais/normas , Ciência dos Animais de Laboratório/normas , Fígado/metabolismo , Masculino , RNA Mensageiro/metabolismo , Ratos/sangue , Ratos/urina , Ratos Sprague-Dawley , Soro/químicaRESUMO
BACKGROUND: Risk analysis of laboratory animal work presupposes allergen monitoring with sensitive methods. Commercial ELISA kits have recently become available for the detection of mouse (Mus m 1) and rat (Rat n 1) urinary allergen from settled dust samples and air samples with high allergen levels. OBJECTIVE: Our aims were to enhance the sensitivities of the commercial ELISA kits for low aeroallergen levels (less than 1 ng/m(3)) and to test these methods with air samples collected from an animal facility. METHODS: Personal and stationary air samples were collected from an animal facility during various tasks of laboratory animal work and from various premises of the animal facility. RESULTS: The sensitivities of the ELISA assays were improved with a careful choice of analysis parameters and reagents. The detection limits of 0.1 ng/m(3) for Mus m 1 and 0.8 ng/m(3) for Rat n 1 were established. The sensitized assays enabled detection of mouse and rat aeroallergens also from premises in which animals or dirty cages were not present during sampling. CONCLUSION: These sensitive assays will help to perform risk assessment in laboratory animal work. However, there remains a lack of standardized analytic procedures and occupational exposure limits for laboratory animal allergens.
Assuntos
Poluentes Ocupacionais do Ar/análise , Alérgenos/urina , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Animais de Laboratório/urina , Pesquisa Biomédica , Camundongos/imunologia , Camundongos/urina , Ratos/imunologia , Ratos/urina , Sensibilidade e EspecificidadeRESUMO
The objective of this report was to derive a simplified approximate estimate of the ion-activity product of calcium oxalate (AP(CaOx)) in rat urine. The relative effect of each urine variable was assessed by means of iterative computerised approximation with the EQUIL2 program. A basic urine composition was chosen from literature and experimental data. The most pronounced influence on AP(CaOx )was recorded for urinary calcium, oxalate, citrate, magnesium and volume. Based on these calculations, an AP(CaOx) index(RAT) was formulated: [formula; see text]. For a 24-h urine sample, factor A takes the value 4067 and factor F should be set to 0.015. Conclusion. A simplified approximate estimate of AP(CaOx) was derived for rat urine. There was a reasonably good correspondence between AP(CaOx) index(RAT) and AP(CaOx), as derived from EQUIL2 ( r=0.890), provided the other urine variables do not deviate very much from that in the basic composition.
Assuntos
Oxalato de Cálcio/química , Oxalato de Cálcio/urina , Modelos Biológicos , Ratos/urina , Animais , ÍonsRESUMO
Interest in the in vivo biological activities of olive oil phenolics is rapidly growing, and different models and vehicles of administration are used worldwide. Matters of practicality determine the use of rats rather than humans as the model of choice. Also, growing interest in nutraceuticals is leading to the formulation of compounds containing olive oil phenols. In this study, we compared metabolism and urinary excretion of hydroxytyrosol [(HT), the most representative phenol of olive oil] between rats and humans by evaluating excretion of HT and its major metabolite, homovanillyl alcohol. Also, we compared human excretion of HT when consumed as a natural component of extra virgin olive oil, when added to refined olive oil, or when added to yogurt (as an approximation of functional food). Urinary excretion of HT was greater in humans than in rats, a species with a high basal excretion of HT and its metabolites. The high (234% of HT administered) excretion of free HT suggests that hydrolysis of oleuropein administered in humans (still an unresolved issue) occurs in vivo. Moreover, human HT excretion was much higher after its administration as a natural component of olive oil (44.2% of HT administered) than after its addition to refined olive oil (23% of HT administered) or yogurt (5.8% of dose or approximately 13% of that recorded after virgin olive oil intake). These data suggest that the rat is not the appropriate model for the study of HT metabolism and that HT-containing functional foods should be carefully formulated.
Assuntos
Antioxidantes/metabolismo , Veículos Farmacêuticos/farmacologia , Álcool Feniletílico/análogos & derivados , Álcool Feniletílico/urina , Ratos/urina , Animais , Combinação de Medicamentos , Humanos , Hidrólise , Glucosídeos Iridoides , Iridoides , Azeite de Oliva , Álcool Feniletílico/administração & dosagem , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Piranos/metabolismo , Especificidade da Espécie , IogurteRESUMO
We have investigated the biological fluids--serum, cerebrospinal fluid, and urine--of three strains of rats; the present data extend our database (also available on-line) and may be of interest for pharmacological and toxicological investigation. Specifically, we have defined reference maps of the major protein components in cerebrospinal fluid and urine. Compartment-specific isoforms were recognized for transferrin and transthyretin. Mass spectrometric data established the cleavage site of the signal peptide and identified the N-terminal blocking group of prostaglandin D synthase from rat cerebrospinal fluid. A previously undescribed member of the family of low molecular mass rat urinary proteins was characterized as containing a sequence similar, but not identical, to the N-terminal region of rat urinary protein-2 (RUP-2), and divergent from RUP-1.
Assuntos
Proteínas Sanguíneas/análise , Proteínas do Líquido Cefalorraquidiano/análise , Bases de Dados de Proteínas , Eletroforese em Gel Bidimensional , Ratos/metabolismo , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Urina/química , Sequência de Aminoácidos , Animais , Feminino , Variação Genética , Internet , Masculino , Dados de Sequência Molecular , Peso Molecular , Isoformas de Proteínas/análise , Proteínas/análise , Proteínas/classificação , Proteinúria/urina , Ratos/sangue , Ratos/líquido cefalorraquidiano , Ratos/urina , Ratos Endogâmicos Lew , Ratos Endogâmicos WKY , Ratos Sprague-Dawley , Especificidade da EspécieRESUMO
This study investigated the direct and indirect effects of male Norway rat (Rattus norvegicus) urine on reproductive, developmental, and fecundity parameters in the dam and her female offspring. Twenty-two dams and litters were studied: 11 in male urine and 11 in distilled water conditions. Only dams were exposed to male urine (or distilled water) from days 14 to 29 postpartum. Significant effects found for the dams exposed to male urine (compared to those only exposed to distilled water) included (i) the second lactational estrus was delayed by 2 days, (ii) vaginal opening and first estrus were 1 day later for female offspring, (iii) the first estrous cycle after vaginal opening was also shorter for their offspring, and (iv) female offspring subsequently produced larger litters than female offspring from dams only exposed to distilled water. Thus, urine from males had direct effects on the timing of the second lactational estrus in dams and indirect effects (mediated by the dam) on developmental and reproductive parameters of her female offspring. Taken as a whole, these results suggest that pheromones in Norway rats may be complex in their effects, context-dependent, and only fully revealed in ecologically relevant contexts. Further study is required to determine whether these effects occur and have biological functions in natural populations.
Assuntos
Animais Recém-Nascidos/crescimento & desenvolvimento , Ratos/urina , Reprodução/fisiologia , Animais , Peso ao Nascer/fisiologia , Estro/fisiologia , Feminino , Tamanho da Ninhada de Vivíparos/fisiologia , Masculino , Ratos Sprague-Dawley , Urina/fisiologiaRESUMO
BACKGROUND: Recent studies in a few industries have shown that the likelihood of IgE-mediated sensitization increases with increasing exposure. The shape of the exposure-response relationships and modification by age, sex, and smoking habit has hardly been studied. OBJECTIVE: The purpose of this study was to determine exposure sensitization relationships for rat sensitization and to evaluate the influence of atopy, smoking habits, and sex. METHODS: Data from 3 cross-sectional studies in The Netherlands, the United Kingdom, and Sweden were used and involved 1062 animal laboratory workers. Selection criteria were harmonized, and this resulted in a study population of 650 animal laboratory workers (60.6% female) with less than 4 years of exposure. Air allergen levels were assessed previously and converted on the basis of an interlaboratory allergen analysis comparison. Available sera were analyzed for the presence of specific antibodies against common allergens (house dust mite, cat, dog, and grass and birch pollen) and work-related allergens (rat and mouse urinary proteins). Questionnaire items on work-related respiratory symptoms, hours worked with rats per week, job performed, smoking habits, and sex were used in this analysis RESULTS: The prevalence of work-related sensitization to rat urinary allergens (IgE >0.7 KU/L) was 9.7 % (n = 63). Thirty-six of the sensitized workers had work-related symptoms (asthma or rhinitis). Two hundred forty-eight workers (38.2%) were atopic (defined as specific IgE to 1 of the common allergens). The sensitization rate increased with increasing air allergen exposure. Atopic workers exposed to low levels of allergen had a more than 3-fold increased sensitization risk compared with nonexposed atopic workers. For atopic subjects, the risk increased little with increasing exposure, whereas for nonatopic subjects, a steadily increasing risk was observed. Smoking and sex did not modify the sensitization risk. CONCLUSION: Rat urinary allergen-sensitization risk increased with increasing exposure intensity. Workers who were atopic had a clearly elevated sensitization risk at low allergen exposure levels.
Assuntos
Alérgenos/urina , Técnicos em Manejo de Animais , Hipersensibilidade/imunologia , Doenças Profissionais/imunologia , Exposição Ocupacional/efeitos adversos , Ratos/imunologia , Alérgenos/imunologia , Animais , Animais de Laboratório/imunologia , Animais de Laboratório/urina , Estudos Transversais , Feminino , Humanos , Hipersensibilidade/diagnóstico , Imunoglobulina E/sangue , Masculino , Camundongos , Ratos/urina , Hipersensibilidade Respiratória , Fatores de Risco , Distribuição por Sexo , FumarRESUMO
Previous studies by the authors demonstrated that the response of urinary aquaporin-2 (AQP2) excretion to dDAVP (deamino-8-D-arginine vasopressin) infusion is an index of vasopressin action on the kidney (N Engl J Med 332: 1540-1545, 1995). In the study presented here, the characteristics of urinary excretion of AQP2 were examined further. An RIA suitable for AQP2 in the urine was established. Relatively high concentrations of detergent and bovine serum albumin in the RIA buffer allowed analysis of urine samples with a wide range of concentrations and increased the sensitivity of the assay. AQP2 in the urine existed as a high molecular weight form of approximately 190 kD by HPLC analysis. The mean urinary AQP2 concentration corrected for creatinine in spot urine samples of healthy subjects who voided in the morning was 1081 +/- 699 fmol/mg creatinine (mean +/- SD, n = 208). The amount of daily excretion of AQP2 in the urine was the same in men and women. Urinary AQP2 content was not affected by age of the subjects and showed a positive correlation with urine osmolality. Finally, the fraction of AQP2 excreted in the urine compared with whole kidney content was determined in the rat. Approximately 3% of AQP2 in the kidney was excreted daily, and this fraction did not change when rats were dehydrated for 3 d. These data demonstrate the necessity of establishing well-designed protocols to use urinary AQP2 as a marker of AVP action.
Assuntos
Aquaporinas , Canais Iônicos/urina , Ratos/urina , Adulto , Idoso , Animais , Aquaporina 2 , Aquaporina 6 , Cromatografia Líquida de Alta Pressão , Desidratação/urina , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Radioimunoensaio , Ratos Sprague-Dawley , Valores de ReferênciaRESUMO
OBJECTIVES: To develop an assay to measure airborne mouse urinary protein (MUP) and to assess the occupational exposure to MUP in the workforce of three establishments as part of an epidemiological study examining the influence of aeroallergen exposure on the development of allergic respiratory disease. METHODS: Personal air samples were collected from nine exposure groups during a workshift. A sensitive and reproducible competitive inhibition assay, which used rabbit antisera specific for MUP, was developed and used to measure the occupational exposure to MUP. RESULTS: The personal measurements of MUP showed that people with direct contact with mice (animal technicians) had the highest exposure followed in decreasing order by those working with anaesthetised animals or their tissue (postmortem workers and scientists) and those with indirect contact with mice (supervisors, office workers, and slide production workers). The only difference in concentrations of MUP between the three establishments were found for cage cleaners, which reflected differences in working practises for this exposure category. Air samples collected during the performance of specific tasks showed that high exposures to MUP were associated with handling mice, indirect contact with mice, and washing floors. CONCLUSIONS: Exposure to mouse urinary proteins has been measured in the occupational environment. This information can be used to determine the relation between exposure to MUP and the development of allergic and respiratory disease.
Assuntos
Poluentes Ocupacionais do Ar/análise , Pessoal de Laboratório Médico , Camundongos/urina , Exposição Ocupacional/análise , Proteínas/análise , Análise de Variância , Animais , Humanos , Técnicas Imunológicas , Ratos/urina , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Surprisingly, many inner-city residents have antibodies to Leptospira interrogans. The manner in which these persons acquire this organism in the absence of recognized occupational, recreational, or epidemic risk factors is not known. OBJECTIVE: To study the epidemiology of patients with leptospirosis who acquired L. interrogans in inner-city Baltimore, Maryland. DESIGN: Epidemiologic investigation. SETTING: Inner-city university hospital. PATIENTS: Three inner-city residents who developed leptospirosis. MEASUREMENTS: Trapping rats in alleys where the patients may have acquired L. interrogans; polymerase chain reaction (PCR) analysis of patients serum and cerebrospinal fluid specimens and rat tissues to determine the presence of leptospiral DNA; and serologic testing of serum from patients and rats by microagglutination assay to confirm L. interrogans infection. RESULTS: Three patients developed leptospirosis after probable percutaneous exposure to rat (Rattus norvegicus) urine in Baltimore alleys. A PCR assay detected L. interrogans DNA in samples of body fluid obtained from the first two patients at presentation (one in cerebrospinal fluid, the other in serum). Results of PCR done on serum drawn from the third patient after antibiotic therapy began were negative. A microagglutination test showed that all patients had high levels of antibodies to the L. interrogans serogroup icterohaemorrhagiae. In 19 of 21 rats that were trapped in the alleys where the patients had sustained lacerations before illness developed, kidney or brain tissues were positive by PCR for the presence of L. interrogans. CONCLUSIONS: A population was discovered to be at risk for acquiring L. interrogans: urban residents who are sporadically exposed to rat urine in the inner city. Inner-city rats often carry L. interrogans. Polymerase chain reaction can quickly establish the diagnosis of leptospirosis and is useful for epidemiologic study. An endemic substrate for the transmission of the organism is present in inner-city Baltimore. Leptospirosis may become increasingly recognized in deteriorating inner cities in which rat populations are expanding.
Assuntos
Leptospirose/epidemiologia , Saúde da População Urbana , Adulto , Animais , Anticorpos Antibacterianos/análise , Baltimore/epidemiologia , DNA Bacteriano/análise , Vetores de Doenças , Feminino , Humanos , Leptospira interrogans/isolamento & purificação , Leptospirose/diagnóstico , Leptospirose/transmissão , Masculino , Reação em Cadeia da Polimerase , Ratos/microbiologia , Ratos/urina , Fatores de RiscoRESUMO
Urine volume (UV), Glomerular filtration rate (GFR) and Fraction of reabsorption (FR) of water, Ca, IP, Na+, K+, Cl-, Glu and UN, which are significant in kidney function analysis, were investigated and compared in three rat strains (WI, SD and F344). UV/body weight ratios were higher in SD rats as compared to other strains. The GFR difference was not significant, but the FRH2O difference was significant in three rat strains. FR of Ca, K+ and Glu showed no significant differences in three different rat strains. SD and F344 were significantly lower than WI in FR of IP. FR of Na+, Cl- and UN were significantly different in three rat strains without Cl- between SD and F344, and UN between WI and SD.
Assuntos
Taxa de Filtração Glomerular/veterinária , Rim/fisiologia , Ratos/fisiologia , Ratos/urina , Animais , Cálcio/urina , Eletrólitos/urina , Glucose/metabolismo , Masculino , Fósforo/urina , Ratos Endogâmicos F344 , Ratos Sprague-Dawley , Ratos Wistar , Ureia/urinaRESUMO
BACKGROUND: In rats, urine has been identified as a major source of the allergens that cause laboratory animal allergy, an important occupational health problem. METHODS AND RESULTS: Urinary proteins and allergens of Wistar rats were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Proteins excreted by male and female rats during puberty were similar and of low molecular weight. In adulthood, moderate increases in diffuse staining of 26 and 21 kd occurred in female urine. In males the 17 kd protein increased dramatically and the 23 and 21 kd proteins moderately. The urine excretion of high-molecular-weight proteins (75 to 63 kd) increased with age in males (females not studied). Immunoblot studies with six sera showed allergens in urine of male and female rats of all ages, three of which were present in all urine (75, 68, and 21 kd). Three allergens (17, 16, and 15 kd) in female urine may be allergenically similar to the 17 kd allergen in adult male urine. Adult male urine allergens were studied further with sera from 83 rat-hypersensitive subjects. Major allergens were identified at 23, 21, and 17 kd, and all sera had IgE to one or more of these proteins. Twenty-seven percent had IgE to 68 and 63 kd allergens. Minor allergens were identified at 75, 51, and 44 kd. CONCLUSIONS: Rat urine is an important source of the major allergens associated with rat hypersensitivity. Age and sex markedly influence the protein and allergenic constituents of rat urine.
Assuntos
Alérgenos/urina , Proteinúria/urina , Ratos/urina , Envelhecimento/fisiologia , Animais , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Immunoblotting , Masculino , Teste de Radioalergoadsorção , Ratos Wistar , Caracteres Sexuais , Dodecilsulfato de SódioRESUMO
The performance of a clinical urinary test-strip reader Clinitek 200 was evaluated for dog and rat urines, in the context of pre-clinical toxicology studies. No major discrepancies were found between data generated by visual estimation or automatic measurement. Analysis of spiked samples showed good agreement between actual concentrations and Clinitek 200 responses for ketone bodies and glucose although a lack of sensitivity was found for the latter. Results for proteins showed over- or underestimation in dog and rat urines respectively at low concentrations, and overestimation at high concentrations in both species. Reproducibility of responses was excellent for ketone bodies, glucose and proteins but was weaker for haemoglobin and bilirubin. High bilirubin concentrations were found to interfere with the haemoglobin reaction. The pH measurements were found to be accurate only around pH 7. Specific gravity measurements were unreliable. Overall, the Clinitek 200 as a screening tool proved sufficiently reliable in the measurement of all parameters tested, with the exception of specific gravity.