RESUMEN
Human cryptosporidiosis remains underdiagnosed, and rapid/accurate diagnosis is of clinical importance. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive, time-consuming, and requires skillful experience. Thus, we aimed to evaluate the usefulness of a coproantigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, we evaluated the performances of a commercial ELISA (CoproELISA Cryptosporidium kit, Savyon Diagnostics, Israel) for the detection of Cryptosporidium spp. in random clinical stool samples through a multicenter study. The sensitivity and specificity for coproantigen ELISA were 98.86% and 94.32%, respectively. The coproantigen ELISA results indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis, and may be useful for large-scale epidemiological studies of cryptosporidiosis.
RESUMEN
Cryptosporidiosis is currently recognized worldwide as a leading cause of moderate to severe diarrhea. In Europe, large water- and foodborne outbreaks have been reported, highlighting the widespread distribution of the parasite and its important health impact. Surveillance networks have been progressively set up and the aim of this study was to present recent epidemiological data obtained in France from 2017 to 2019 by the National Reference Center-Expert Laboratory of cryptosporidiosis (Centre National de Référence-Laboratoire Expert cryptosporidioses CNR-LE). Data were obtained from online reports of volunteer network participants and stools were sent to the CNR-LE for species identification and GP60 genotyping. During this period, data from 750 online reports were available. Cryptosporidiosis occurred predominantly in young children (<5 years old) and in young adults, especially during late summer. Most patients were immunocompetent (60%), and deaths were reported only in immunocompromised patients. Cryptosporidium parvum was largely predominant (72% of cases) over C. hominis (24%) and some other uncommon species. C. parvum GP60 subtypes IIa and IId were the most represented, which suggests frequent zoonotic transmission. For C. hominis, subtypes IbA10G2 and IaA22R2 were predominant.
Asunto(s)
Parasitosis Intestinales , Parásitos , Refugiados , Animales , Estudios Transversales , Heces , Humanos , Parasitosis Intestinales/diagnóstico , ItaliaRESUMEN
Purpose/Aims: Infectious keratitis is a major cause of visual impairment and blindness worldwide. Common difficulties in treating fungal keratitis prompt new therapeutic possibilities. In this study, intrastromal voriconazole and posaconazole, and topical posaconazole were tested for their potential to obtain therapeutic cornea concentrations. Materials and Methods: Pharmacokinetics of triazole intracorneal/eye drop administration was studied in rats. Sixty-two rats were treated either by voriconazole or posaconazole. Twenty-nine and 33 rats received intrastromal injection of voriconazole solution (1 µl, 10 mg/ml) and posaconazole solution (1 µl, 18 mg/ml), respectively, administered under microscopic examination with a 32 gauge needle in the left cornea. Posaconazole (1.8% solution) eye drops were used. Cornea and plasma concentrations were determined using 2D HPLC separation and tandem MS, at 30 min, 3 h, 6 h, 24 h, 48 h, 72 h, and 144 h (6 days) post-intrastromal injection. The entire rat cornea was used for chromatography analyses. Results: In anesthetized rats, single intracorneal injection resulted, after 30 min, in respectively, >300 ng/mg and >260 ng/mg cornea concentrations, dropping to low levels within hours, while staying low in plasma. The effect of hourly posaconazole eye drops resulted in >10 ng/mg cornea concentration, which was maintained with instillations every 2 and then every 4 h. Conclusion: Our results show that there is little interest of intrastromal triazole administration due to the short duration of high cornea concentrations obtained after intracorneal injection. Posaconazole eye drops maintain therapeutic cornea concentrations in rats and could be used to treat severe infectious keratitis.
Asunto(s)
Antifúngicos/farmacocinética , Córnea/metabolismo , Triazoles/farmacocinética , Voriconazol/farmacocinética , Administración Oftálmica , Animales , Antifúngicos/administración & dosificación , Cromatografía Liquida , Sustancia Propia/efectos de los fármacos , Inyecciones Intraoculares , Masculino , Pruebas de Sensibilidad Microbiana , Soluciones Oftálmicas , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem , Voriconazol/administración & dosificaciónRESUMEN
BACKGROUND: Effective stool concentration is essential in microscopically based diagnosis of human intestinal parasite infections. OBJECTIVE: To compare the performances of 4 concentration commercial kits and 1 homemade procedure in 96 clinical stool specimens that tested positive for the detection of 9 helminth and 8 protozoa parasites. METHODS: The presence or absence of parasite forms was microscopically determined under conditions of standard practice. Also, we established the accuracies, concentration factors, and extraction yields. RESULTS: No difference was observed between procedures for preconcentration specimens that tested positive. However, for preconcentration specimens that tested negative, we discovered that the homemade procedure was the most effective, and 2 of the 4 commercial kits were discovered to be satisfactory for routine applications. CONCLUSIONS: For all parasites, procedures with biphasic solvents exhibited higher performances than organic solvent-free procedures. For the first time, the effectiveness of commercial concentration kits has been evaluated on several common stool parasites, and the results suggest that improvement of commercial procedures is possible.
Asunto(s)
Técnicas de Laboratorio Clínico/métodos , Pruebas Diagnósticas de Rutina/métodos , Heces/parasitología , Parasitosis Intestinales/diagnóstico , Parásitos/aislamiento & purificación , Manejo de Especímenes/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
[This corrects the article DOI: 10.1371/journal.pone.0194058.].
RESUMEN
BACKGROUND: Cryptosporidium spp. infections are the most frequent parasitic cause of diarrhea in humans and cattle. However, asymptomatic cases are less often documented than symptomatic cases or cases with experimentally infected animals. Cryptosporidium (C.) hominis infection accounts for the majority of pediatric cases in several countries, while C. parvum is a major cause of diarrhea in neonatal calves. In cattle Cryptosporidium spp. infection can be caused by C. parvum, C. bovis, C.andersoni and C. ryanae, and recently, reports of cattle cases of C. hominis cryptosporidiosis cases suggest that the presence of C. hominis in calves was previously underestimated. METHODOLOGY/PRINCIPAL FINDINGS: From February to November 2015, Cryptosporidium spp. infected calves were detected in 29/44 randomly included farms from 5 geographic regions of France. C. hominis and C. parvum were found in 12/44 and 26/44 farms, respectively with higher C. hominis prevalence in the western region. In 9 farms, both C. parvum and C. hominis were detected. Eighty-six of 412 (73/342 asymptomatic and 13/70 symptomatic) one to nine-week-old calves shed C. hominis or C. parvum oocysts (15 and 71 calves, respectively), with no mixed infection detected. The predominant C. hominis IbA9G3 genotype was present in all regions, and more frequent in the western region. An incompletely characterized Ib, and the IbA13G3, IbA9G2 and IbA14G2 genotypes were present only in the western region. For C. parvum, the most frequent genotype was IIaA16G3R1 with no geographic clustering. Most C. hominis infected calves were asymptomatic, with some exceptions of IbA9G2 and IbA9G3 isolates, while C. parvum IIaA16G3R1 was associated with symptoms. CONCLUSIONS/SIGNIFICANCE: Present results indicate for the first time that in several geographic regions of France, C. hominis was present in about one fifth of both asymptomatic and symptomatic infected calves, with isolated genotypes likely associated with human infection. Further investigations are aimed at documenting direct or indirect transmissions between livestock and humans.
Asunto(s)
Infecciones Asintomáticas/epidemiología , Enfermedades de los Bovinos/epidemiología , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Cryptosporidium/fisiología , Animales , Animales Recién Nacidos/parasitología , Bovinos , Enfermedades de los Bovinos/parasitología , Criptosporidiosis/transmisión , Cryptosporidium/genética , Cryptosporidium/aislamiento & purificación , Diarrea/epidemiología , Diarrea/parasitología , Diarrea/veterinaria , Heces/parasitología , Francia/epidemiología , Genotipo , Humanos , Oocistos/aislamiento & purificación , PrevalenciaRESUMEN
Background: Acanthamoeba keratitis (AK) is a sight-threatening infectious disease. Its effective and safe medical therapy remains highly debated. Recently, voriconazole, a monotriazole with noted in vitro activity against a large variety of fungi, has been successfully used both topically and systemically to treat human AK cases. Objectives: To measure anti-Acanthamoeba polyphaga in vitro activity, anti-rat AK efficiency and rat cornea penetration of eye-drop and oral voriconazole. Methods: A. polyphaga was maintained in axenic cultures. In vitro, amoebicidal and cysticidal activities of voriconazole were measured using an XTT assay. AK lesions of Sprague Dawley rats were scored from grade 0 to grade 3. For 21 days, from day 7 post-infection, voriconazole (1% solution) eye drops were instilled or voriconazole was administered by gavage (60 mg/kg/day). After killing, superficial corneal epithelium scrapings were cultured and analysed by PCR, and eye-globe histology was performed. Cornea and plasma concentrations were determined using 2D HPLC separation and tandem MS. Results: In vitro, voriconazole inhibited trophozoite proliferation with an IC50 value of 0.02 mg/L and an IC90 value of 2.86 mg/L; no cysticidal effect was found. In AK rats, eye drops reduced clinical worsening from day 7 to day 14 post-infection and oral voriconazole was not effective. Voriconazole cornea concentrations were directly dependent on the frequency of eye-drop instillations, which resulted in lower plasma concentrations, whilst oral voriconazole resulted in lower cornea concentrations. Conclusions: Present data underline the need for high-frequency eye-drop instillation regimens for efficient AK therapy.
Asunto(s)
Queratitis por Acanthamoeba/tratamiento farmacológico , Acanthamoeba/efectos de los fármacos , Antiprotozoarios/farmacología , Córnea/efectos de los fármacos , Voriconazol/farmacología , Acanthamoeba/genética , Administración Oral , Animales , Antiprotozoarios/administración & dosificación , Cultivo Axénico , Córnea/parasitología , Masculino , Pruebas de Sensibilidad Microbiana , Soluciones Oftálmicas/administración & dosificación , Soluciones Oftálmicas/farmacología , Ratas , Ratas Sprague-Dawley , Voriconazol/administración & dosificaciónRESUMEN
Somatostatins are proteins that are involved in gastrointestinal function. However, little is known with regard to somatostatin receptor subtype (SSTR) expression changes that occur in the jejunum during low-grade inflammation and during subsequent octreotide treatment. The aim of the present study was to investigate the expression of SSTRs in the jejunums of Cryptosporidium parvum (C. parvum)-infected rats by immunohistochemisty, reverse transcription (RT) PCR and quantitative real-time RT-PCR assays. Five-day-old suckling Sprague-Dawley rats (n = 15 for each group) were orally gavaged with 105 Nouzilly isolate (NoI) oocysts. Rats then received 50 µg/kg/day of octreotide by intraperitoneal injection from day 10 to day 17 post-infection. Animals were sacrificed on days 7 and 14 post-infection for immunohistochemical analysis and on days 14, 35 and 50 for mRNA expression analysis of SSTR subtypes. Histological analysis of jejunum tissues demonstrated infection of C. parvum along the villus brush border on day 7 post-infection and infection clearance by day 14 post-infection. Real-time PCR analysis indicated that in the inflamed jejunum, a significant increase in SSTR1 and SSTR2 expression was observed on day 14 post-infection. Octreotide therapy down-regulated the expression of SSTR2 on day 37 post-infection but significantly increased expression of SSTR1, SSTR2 and SSTR3 on day 50 post-infection. The results indicate that specific SSTRs may regulate the inflammatory pathway in the rat intestinal inflammation model.
Asunto(s)
Antiinflamatorios/farmacología , Criptosporidiosis/metabolismo , Cryptosporidium parvum , Regulación de la Expresión Génica/efectos de los fármacos , Enfermedades del Yeyuno/metabolismo , Yeyuno/efectos de los fármacos , Octreótido/farmacología , Receptores de Somatostatina/biosíntesis , Animales , Animales Lactantes , Antiinflamatorios/administración & dosificación , Criptosporidiosis/patología , Inflamación , Mucosa Intestinal/metabolismo , Enfermedades del Yeyuno/patología , Yeyuno/metabolismo , Yeyuno/patología , Octreótido/administración & dosificación , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Somatostatina/genéticaRESUMEN
The aim of this work was to document molecular epidemiology of Rasamsonia argillacea species complex isolates from cystic fibrosis (CF) patients. In this work, 116 isolates belonging to this species complex and collected from 26 CF patients and one patient with chronic granulomatous disease were characterized using PCR amplification assays of repetitive DNA sequences and electrophoretic separation of amplicons (rep-PCR). Data revealed a clustering consistent with molecular species identification. A single species was recovered from most patients. Rasamsonia aegroticola was the most common species, followed by R. argillacea sensu stricto and R. piperina, while R. eburnea was not identified. Of 29 genotypes, 7 were shared by distinct patients while 22 were patient specific. In each clinical sample, most isolates exhibited an identical genotype. Genotyping of isolates recovered from sequential samples from the same patient confirmed the capability of R. aegroticola and R. argillacea isolates to chronically colonize the airways. A unique genotype was recovered from two siblings during a 6-month period. In the other cases, a largely dominant genotype was detected. Present results which support the use of rep-PCR for both identification and genotyping for the R. argillacea species complex provide the first molecular evidence of chronic airway colonization by these fungi in CF patients.
Asunto(s)
Fibrosis Quística/complicaciones , Eurotiales/clasificación , Eurotiales/aislamiento & purificación , Micosis/diagnóstico , Micosis/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Análisis por Conglomerados , Electroforesis , Eurotiales/genética , Genotipo , Humanos , Técnicas Microbiológicas/métodos , Epidemiología Molecular , Micosis/microbiología , Secuencias Repetitivas de Ácidos Nucleicos/genéticaRESUMEN
PURPOSE: The Scedosporium apiospermum species complex usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF), but little is known about the molecular epidemiology of the airway colonization. METHODS: Polymerase chain reaction (PCR) amplification of repetitive sequences (rep-PCR) was applied to the retrospective analysis of a panel of isolates already studied by random amplification of polymorphic DNA (RAPD) and comprising 63 isolates recovered from sputa from 9 CF patients. Results were compared to those obtained previously by RAPD, and herein by beta-tubulin (TUB) gene sequencing and Multilocus Sequence Typing (MLST). RESULTS: Within the panel of isolates studied,S. apiospermum sensu stricto and Scedosporium boydii, as expected, were the predominant species with 21 and 36 isolates, respectively. Four isolates from one patient were identified as Scedosporium aurantiacum, whereas two isolates belonged to the Pseudallescheria ellipsoidea subgroup of S. boydii rep-PCR analysis of these isolates clearly differentiated the three species and P. ellipsoidea isolates, whatever the rep-PCR kit used, and also permitted strain differentiation. When using the mold primer kit, results from rep-PCR were in close agreement with those obtained by MLST. For both S. apiospermum and S. boydii, 8 genotypes were differentiated by rep-PCR and MLST compared to 10 by RAPD. All S. aurantiacum isolates shared the same RAPD genotype and exhibited the same rep-PCR profile and sequence type. CONCLUSIONS: These results illustrate the efficacy of rep-PCR for both species identification within the S. apiospermum complex and genotyping for the two major species of this complex.Abstract presentation: Part of this work was presented during the 18th Congress of the International Society for Human and Animal Mycology, Berlin (Germany), June 2012.S. Giraud, C. Godon, A. Rougeron, J.P. Bouchara and L. Favennec are members of the ECMM/ISHAM working group on Fungal respiratory infections in Cystic Fibrosis(Fri-CF).
Asunto(s)
Tipificación Molecular/métodos , Micosis/microbiología , Reacción en Cadena de la Polimerasa/métodos , Técnica del ADN Polimorfo Amplificado Aleatorio/métodos , Scedosporium/genética , Fibrosis Quística/microbiología , Humanos , Filogenia , Scedosporium/clasificación , Esputo/microbiologíaRESUMEN
The intestinal protozoan Giardia duodenalis includes 2 genetically distinct assemblages, A and B, which are responsible for human infections. Little is known so far on the genotypes of G. duodenalis human isolates in France. The present characterization of 19 French clinical isolates was aimed at determining their genotype patterns and associations with clinical symptoms, and in vivo metronidazole resistance, respectively. Based on both triose-phosphate isomerase (tpi) and ß-giardin (bg) gene sequences, twelve isolates were identified as assemblage A, and 7 as assemblage B for the 2 gene loci. Sub-genotyping heterogeneities were observed in 15/19 isolates attributed to either A or B assemblage. They include frequent mismatches and intra-assemblage discordances and mixed positions, which were found more frequently in tpi than in bg sequences, and in assemblage B than in assemblage A sequences. No association was found between sub-genotypes, clinical symptoms and metronidazole sensitivity. Present data underline the need for improvements in the standardization of G. duodenalis multilocus genotyping approach for further molecular epidemiologic studies of giardiasis.
Asunto(s)
Proteínas del Citoesqueleto/genética , Giardia lamblia/clasificación , Giardiasis/parasitología , Proteínas Protozoarias/genética , Triosa-Fosfato Isomerasa/genética , Adulto , Animales , Secuencia de Bases , Niño , Preescolar , ADN Protozoario/química , ADN Protozoario/genética , Heces/parasitología , Francia , Genotipo , Giardia lamblia/genética , Giardia lamblia/aislamiento & purificación , Humanos , Lactante , Ratones , Datos de Secuencia Molecular , Tipificación de Secuencias Multilocus , Filogenia , Análisis de Secuencia de ADNRESUMEN
Thirty-nine new thiazolide/thiadiazolide compounds were compared with the nitrothiazole nitazoxanide for activity against Cryptosporidium parvum development in HCT-8 cells. Twenty-seven agents exerted > or =90% inhibition. Agents with a lower 50% inhibitory concentration (IC(50)) than nitazoxanide were either NO(2) or halogen 5 substituted on the thiazole moiety. Other 5 substitutions such as methyl, C(3)H(7), C(6)H(11), H, SO(2)CH(3), and SCH(3) negatively impacted activity. Five-substituted deacetylated analogues exhibited higher IC(50)s than their acetylated counterparts. Halogeno-thiazolide/thiadiazolides may provide valuable nitro-free alternatives to nitazoxanide.
Asunto(s)
Cryptosporidium parvum/efectos de los fármacos , Tiadiazinas/farmacología , Tiazoles/farmacología , Antiprotozoarios/farmacología , Cryptosporidium parvum/crecimiento & desarrollo , Humanos , Concentración 50 Inhibidora , Nitrocompuestos , Pruebas de Sensibilidad Parasitaria , Relación Estructura-Actividad , Tiadiazinas/química , Tiazoles/químicaRESUMEN
Cryptosporidium spp. are a cause of self-limited diarrhea in immunocompetent hosts. In immunocompetent rats, Cryptosporidium parvum infection induced digestive hypersensitivity, a key pathophysiological factor in functional digestive disorders such as irritable bowel syndrome (IBS). In such a rat model, we sought to document whether jejunal hypersensitivity depends on C. parvum isolate and is associated with a mast cell accumulation. Five-day-old rats were orally administered 10(5) oocysts of either Nouzilly (NoI) or Iowa (IoI) C. parvum isolate. NoI-infected rats exhibited the lowest food intake on days 7 and 14 postinfection (p.i.). On day 7 p.i., small intestine villus atrophy, crypt hyperplasia, and inflammatory cell infiltration were prominent in NoI-infected rats, with higher numbers of Cryptosporidium forms than in IoI-infected rats. Compared to uninfected control rats, jejunal intraepithelial lymphocytes (IELs) were increased only in NoI-infected rats on day 14 p.i. On day 50 p.i., jejunal hypersensitivity to distension was found only in NoI-infected rats; this hypersensitivity is associated with activated mast cell accumulation. The number of mast cells in the jejunal lamina propria was increased from day 36 p.i. in NoI-infected rats and only at day 120 p.i. in IoI-infected rats. Our data suggest that both the severity of infection (weight loss, reduced food intake, villus atrophy, and IEL accumulation) and the onset of a jejunal hypersensitivity after infection in association with an activated mast cell accumulation are isolate dependent and related to NoI infection. This cryptosporidiosis rat model is a relevant model for the study of underlying mechanisms of postinfectious IBS-like symptoms.
Asunto(s)
Criptosporidiosis/inmunología , Criptosporidiosis/fisiopatología , Cryptosporidium parvum/genética , Yeyuno/microbiología , Mastocitos/inmunología , Animales , Criptosporidiosis/patología , Cryptosporidium parvum/inmunología , Cryptosporidium parvum/patogenicidad , Modelos Animales de Enfermedad , Inmunohistoquímica , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Yeyuno/inmunología , Yeyuno/patología , Umbral del Dolor , RatasRESUMEN
OBJECTIVE: To investigate the relation between the adrenal production of gluco- and mineralocorticoids, the inflammatory status and the outcome in critically ill patients with liver cirrhosis. DESIGN: Prospective descriptive study. SETTING: Medical intensive care unit (ICU) in a university hospital. PATIENTS: Fifty consecutive patients with liver cirrhosis. INTERVENTIONS: A corticotropin stimulation test within 12h following ICU admission. Plasma cortisol concentration was measured before and after the test. Renin and aldosterone concentrations, as well as interleukin-6 (IL-6) level to assess the pro-inflammatory status, were measured only before the test. Impaired adrenal function was defined as cortisol response to the test less than 9microg/dl. Hyperreninemic hypoaldosteronism syndrome was defined as basal renin over aldosterone ratio (RRA) higher than 2. MEASUREMENTS AND RESULTS: Forty-one (82%) patients had impaired adrenal function, and 26 patients (52%) presented with RRA > 2. Patients with RRA > 2 exhibited greater disease severity and organ dysfunction scores at baseline, higher levels of serum renin and IL-6, and a greater ICU mortality rate, but risk-adjusted mortality rates were not different between the two groups. Renin and IL-6 plasma concentrations were positively correlated. Finally, in a Cox regression analysis, independent predictors of 30-day mortality were hyperreninemic hypoaldosteronism syndrome, IL-6 higher than 400pg/ml and severe renal failure. CONCLUSIONS: Adrenal dysfunction was common in critically ill cirrhotic patients. Hyperreninemic hypoaldosteronism syndrome was related to a greater pro-inflammatory status and degree of acute organ failure, and was independently associated with a worse prognosis.
Asunto(s)
Enfermedad Crítica , Hipoaldosteronismo/metabolismo , Interleucina-6/análisis , Cirrosis Hepática/fisiopatología , Síndrome , Anciano , Estudios de Cohortes , Femenino , Glucocorticoides/biosíntesis , Humanos , Unidades de Cuidados Intensivos , Interleucina-6/sangre , Interleucina-6/metabolismo , Cirrosis Hepática/inmunología , Masculino , Persona de Mediana Edad , Mineralocorticoides/biosíntesis , Estudios Prospectivos , Sistema Renina-AngiotensinaRESUMEN
A cohort of 10 Hashimoto's encephalopathy (HE) patients, 33 patients with unrelated neurological symptoms, 12 Hashimoto's thyroiditis patients and 4 healthy adult donors was studied to explore the neurological targets of anti-thyroperoxidase (TPO) autoantibodies (aAb) in HE. High levels of anti-TPO aAb were only detected in HE group's cerebrospinal fluids. In immunofluorescence assays on monkey brain cerebellum sections, both HE patients' sera and anti-TPO monoclonal antibodies (mAb) were able to bind cerebellar cells expressing glial fibrillary acid protein. Normal human astrocytes from primary cultures also reacted with anti-TPO mAb. Specific astrocyte binding of anti-TPO aAb suggests a role of these aAb in the HE pathogenesis.
Asunto(s)
Astrocitos/metabolismo , Autoanticuerpos/análisis , Cerebelo/patología , Enfermedad de Hashimoto/inmunología , Yoduro Peroxidasa/inmunología , Tiroglobulina/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Autoanticuerpos/inmunología , Células Cultivadas , Cerebelo/metabolismo , Femenino , Feto , Proteína Ácida Fibrilar de la Glía/metabolismo , Enfermedad de Hashimoto/patología , Humanos , Yoduro Peroxidasa/sangre , Yoduro Peroxidasa/líquido cefalorraquídeo , Masculino , Persona de Mediana Edad , Radioinmunoensayo , Tiroglobulina/sangre , Tiroglobulina/líquido cefalorraquídeo , Glándula Tiroides/metabolismo , Glándula Tiroides/patología , Factores de TiempoRESUMEN
AIM: To quantify the intraepithelial lymphocytes (IELs) and to document the membrane expression of CD4, CD8, TCRgamma delta and adhesion and/or activation-associated molecules (CD103, CD28, CD44, CD69, HLA-DR, CD95/Fas) in the duodenal mucosa of patients with functional dyspepsia (FD) in order to provide arguments for an immunological process in FD. METHODS: Twenty-six FD patients according to Rome II criteria (20 were H pylori negative) were studied and compared to 12 healthy adults. IELs were isolated from five duodenal biopsy samples, then quantified by microscopy and flow cytometry while the membrane phenotypes were determined by cytofluorometry. RESULTS: Duodenal histological examination was normal. In H pylori negative patients, the number of IELs was not different from that in healthy controls. Median percentage expression of CD4, CD8, or TCRgamma delta and CD103, CD44, CD28, CD69 on CD3+ IELs, among the adhesion/activation associated molecules tested, was not different from that in healthy controls. In contrast, the median percentage expression of CD95/Fas [22 (9-65) vs 45 (19-88), P=0.03] and HLA-DR expressing CD3+ IELs [4 (0-30) vs 13 (4-42), P=0.04] was significantly lower in the H pylori negative FD group than in healthy controls, respectively. The number of IELs was significantly greater in H pylori positive FD patients than in healthy controls [median ratio for 100 enterocytes 27.5 (6.7-62.5) vs 10.8 (3-33.3), P=0.02] due to a higher number of CD8+ CD3+ IELs. CONCLUSION: In H pylori negative FD patients, the phenotypic characterization of IELs suggests that we cannot exclude a role of IELs in FD.
Asunto(s)
Duodeno/inmunología , Dispepsia/inmunología , Mucosa Intestinal/patología , Linfocitos T/metabolismo , Adolescente , Adulto , Anciano , Biopsia , Complejo CD3/metabolismo , Antígenos CD8/metabolismo , Estudios de Casos y Controles , Moléculas de Adhesión Celular/metabolismo , Recuento de Células , Membrana Celular/inmunología , Duodeno/metabolismo , Duodeno/patología , Dispepsia/metabolismo , Femenino , Humanos , Mucosa Intestinal/metabolismo , Masculino , Persona de Mediana Edad , Fenotipo , Estudios ProspectivosRESUMEN
In 5-day-old immunocompetent Sprague-Dawley rats infected with either 10(2) or 10(5) Cryptosporidium parvum oocysts, transient infection resulted 120 days later in increased cardiovascular depressor response to jejunal distension and jejunal myeloperoxidase activity (P < 0.05). Nitazoxanide treatment normalized jejunal sensitivity (P < 0.001) but not myeloperoxidase levels (P > 0.05). Data warrant further evaluation of the role of early cryptosporidiosis in the development of chronic inflammatory gut conditions.
Asunto(s)
Criptosporidiosis/inmunología , Cryptosporidium parvum , Hipersensibilidad/inmunología , Yeyuno/inmunología , Animales , Animales Recién Nacidos , Criptosporidiosis/fisiopatología , Femenino , Hipersensibilidad/parasitología , Hipersensibilidad/fisiopatología , Yeyuno/parasitología , Yeyuno/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
PURPOSE: To develop a rat model of chronic Acanthamoeba polyphaga keratitis suitable for pharmacologic assessment of therapeutic agents. METHODS: An A. polyphaga isolate (ATCC #50495) was grown in peptone-yeast extract-glucose medium. Five-weeks-old, Sprague-Dawley male rats were injected with 10(3) or 10(4) trophozoites in the left cornea stromal layer. A subconjunctival injection of 0.14, 0.28, or 0.57 mg long-acting betamethasone was performed weekly. At the end of experiments, rats were killed; the superficial corneal epithelium gently scraped and cultured; and globes histologically examined. Topical polyhexamethylene biguanide (PHMB), hexamidine diisethionate, and miltefosine (hexadecylphosphocholine) were administered topically as eye drops 3 times a day at concentrations of 0.02%, 0.1%, and 0.01% respectively. In vitro minimal inhibitory concentration (MIC) and fractional inhibitory concentration values were measured in A. polyphaga cultures. RESULTS: In infected eyes, lesions consisted of the sequential appearance within 2 weeks of edema, infiltrates, and/or abscesses. On day 35 postinfection, a combination of 10(4) parasites with a regimen of 0.28 mg/week betamethasone resulted in the highest ratio of rats with abscesses. Presence of A. polyphaga was confirmed histologically and inconsistently in cultures. In rats optimally prepared as said earlier, agents were administered on day 6 postinfection. A combination of PHMB and hexamidine diisethionate exerted a synergistic effect and was more effective than PHMB, hexamidine diisethionate, or miltefosine alone. In vitro, PHMB (MIC = 14.6 microM) and hexamidine diisethionate (MIC = 555 microM) exerted a synergistic effect (fractional inhibitory concentration = 0.06), and miltefosine exhibited antiamoebal activity (MIC = 27.4 microM). CONCLUSIONS: In this study, a rat model of chronic A. polyphaga keratitis was obtained and found suitable for assessment of pharmacologic agents. It provides an in vivo approach of drug resistance, pathogenicity, and physiopathologic mechanisms of chronic amoebic keratitis.
Asunto(s)
Queratitis por Acanthamoeba/tratamiento farmacológico , Antiprotozoarios/uso terapéutico , Betametasona/uso terapéutico , Modelos Animales de Enfermedad , Glucocorticoides/uso terapéutico , Acanthamoeba/efectos de los fármacos , Acanthamoeba/aislamiento & purificación , Queratitis por Acanthamoeba/parasitología , Queratitis por Acanthamoeba/patología , Administración Tópica , Animales , Benzamidinas/uso terapéutico , Biguanidas/uso terapéutico , Enfermedad Crónica , Sustancia Propia/efectos de los fármacos , Sustancia Propia/parasitología , Sustancia Propia/patología , Evaluación de Medicamentos/métodos , Quimioterapia Combinada , Masculino , Pruebas de Sensibilidad Parasitaria , Fosforilcolina/análogos & derivados , Fosforilcolina/uso terapéutico , Ratas , Ratas Sprague-Dawley , Organismos Libres de Patógenos EspecíficosRESUMEN
Rhodococcus equi infection is considered the most common cause of pneumonia in foals less than 6 months of age. Immunization of foals and/or mares may become a procedure of choice for prevention. The present work documents the antibody response of neonate foals to R. equi virulence-associated protein (Vap) vaccine candidate peptides. A mixture of 4 R. equi (ATCC 33701) Vap peptides was selected based on their hydrophilicity and recognition by naturally acquired IgG antibodies from 13 adult horses and 33 neonate foals from France and Japan. They were combined with a water-based nanoparticular adjuvant to promote a protective immune response including both Th1 cytokine pattern and antibody response. A single intramuscular injection resulted in an IgG antibody response 30 days later, although inconsistently. In responding animals, no bias in IgG subclass distribution was observed, and antibody response was associated with enhanced serum opsonic activity. In conclusion, data indicate that synthetic Vap peptides combined with nanoparticular adjuvant were immunogenic and resulted in a significant increase in IgG antibodies against the corresponding virulent R. equi strain in a majority of foals.