Performance of a real time PCR for leishmaniasis dignosis using a L. (L.) infantum hypothetical protein as target in canine samples

Visceral leishmaniasis represents an important public health issue in different parts of the world, requiring that measures be put in place to control the spread of the disease worldwide. The canine leishmaniasis diagnosis is not easy based on clinical signs, since dogs may not develop the infection with recognizable signs. Thus, the laboratorial diagnosis is essential to ascertain the incidence and prevalence of canine leishmaniasis especially in areas with major control efforts. Although, the diagnosis can be performed by the use of different approaches, the molecular methods such as PCR have become an indispensable tool for leishmaniases diagnosis. A TaqMan assay for real-time PCR (Linj31-qPCR) was developed to determine the parasite occurrence in clinical cases of leishmaniasis. The assay targets an L. (L.) infantum hypothetical protein region. The specificity of the assay was verified by using Leishmania World Health Organization reference strains including parasites belonging to subgenus L. (Leishmania), subgenus L. (Viannia), other Leishmania species and Trypanosoma cruzi. The sensitivity was verified by using isolates of L. (L.) amazonensis and L. (L.) infantum. The usefulness of the assay for diagnosis was ascertained by testing 277 samples from dogs in regions endemic for visceral and/or cutaneous leishmaniasis and from regions in which leishmaniasis was not endemic in São Paulo State, Brazil. Diagnosis of canine visceral leishmaniasis (CVL) was determined on these animals by conventional PCR and three serological tests. The dog samples were divided into four groups. I, dogs with CVL (n = 101); II, dogs with other diseases and without CVL (n = 97); III, dogs with American cutaneous leishmaniasis (n = 7), and, IV, dogs without CVL (n = 72) from areas where leishmaniasis was not endemic as control group. Results indicated that Linj31-qPCR was able to identify parasites belonging to subgenus L. (Leishmania) with no cross-amplification with other parasite subgenera. The Linj31-qPCR detected Leishmania parasites DNA in 98% of samples from Group I. In conclusion this methodology can be used as routine diagnostic tools to detect parasites from subgenus Leishmania.

Cerebral and ocular toxoplasmosis related with IFN-y, TNF-a, and IL-10

This study analyzed the synthesis of Interferon gamma (IFN-γ), Tumor Necrosis Factor alpha (TNF-α), and Interleukin 10 (IL-10) in chronically infected patients which developed the symptomatic disease as cerebral or ocular toxoplasmosis. Blood from 61 individuals were divided into four groups: Cerebral toxoplasmosis/AIDS patients (CT/AIDS group) (n = 15), ocular toxoplasmosis patients (OT group) (n = 23), chronic toxoplasmosis individuals (CHR group) (n = 13) and healthy individuals (HI group) (n = 10). OT, CHR, and HI groups were human immunodeficiency virus (HIV) seronegative. The diagnosis was made by laboratorial (PCR and ELISA) and clinical subjects. For cytokine determination, peripheral blood mononuclear cells (PBMC) of each patient were isolated and stimulated in vitro with T. gondii antigen. IFN-γ, TNF-α, and IL-10 activities were determined by ELISA. Patients from CT/AIDS and OT groups had low levels of IFN-γ when were compared with those from CHR group. These data suggest the low resistance to develop ocular lesions by the low ability to produce IFN-γ against the parasite. The same patients, which developed ocular or cerebral toxoplasmosis had higher TNF-α levels than CHR individuals. High TNF-α synthesis contribute to the inflammatory response and damage of the choroid and retina in OT patients and in AIDS patients caused a high inflammatory response as the TNF-α synthesis is not affected since monocytes are the major source this cytokine in response to soluble T. gondii antigens. IL-10 levels were almost similar in CT/AIDS and OT patients but low when compared with CHR individuals. The deviation to Th2 immune response including the production of anti-inflammatory cytokines, such as IL-10 may promote the parasite's survival causing the tissue immune destruction. IL-10 production in T. gondii-infected brains may support the persistence of parasites as down-regulating the intracerebral immune response. All these indicate that OT and CT/AIDS patients produced low levels of IL-10 (Th2 response) and IFN-γ (Th1 response). They produced high TNF-α suggesting a high inflammatory response triggered by the parasite.

Anillos de reconstrucción en la cirugía de revisión de las artroplastias de cadera / Acetabular rings in revision total hip replacement

Objetivo. Evaluar los resultados radiográficos de los anillos de reconstrucción utilizados en la cirugía de revisión de grandes defectos acetabulares. Material y método. Se ha estudiado la evolución radiológica de 13 anillos de reconstrucción utilizados en grados 3 y 4, con un seguimiento mínimo de 25 meses y máximo de 60 (media de seguimiento: 37 meses). En todos los casos se utilizaron aloinjertos debajo del anillo y cúpula cementada. Las mediciones radiológicas usadas tomaban como referencia la posición, altura y profundidad del anillo, así como la evolución del centro de la cabeza protésica. También se registró la presencia de complicaciones y la recuperación de la dismetría de extremidades. Resultados. Se presentaron tres complicaciones, una lesión vascular intraoperatoria, una luxación posterior y una infección superficial. El descenso medio del centro de rotación fue de 4 cm. En ningún caso fue necesario reintervenir a los pacientes. La dismetría de extremidades disminuyó de 6,3 cm en el preoperatorio a 3,1 cm en el postoperatorio inmediato. Después no se encontraron modificaciones en las mediciones radiográficas. Conclusiones. La utilización de un anillo de reconstrucción en la cirugía de revisión acetabular permite descender el centro de rotación de la cadera y recuperar precozmente la estructura y depósito óseo sin complicaciones graves (AU)

Purpose. To assess the radiographic results of reconstruction rings used in surgical procedures conducted to revise large acetabular defects. Materials and methods. A study was carried out to analyze the radiological evolution of 13 acetabular rings used in grade 3 and 4 patients, with a follow-up between 25 and 60 months (mean: 37 months). In all cases, allografts were placed underneath the ring and the cemented cup. The radiological measurements taken used as their landmarks the position, height and depth of the ring as well as the evolution of the center of the prosthetic head. The appearance of complications was also recorded, as was the restoration of limb dysmetry. Results. There were three complications, an intraoperative vascular lesion, a posterior dislocation as well as a superficial infection. The average downward migration of the center of rotation was 4 cm. In no case was it necessary to reoperate. Limb dysmetry went down from a preop value of 6.3 cm to 3.1 at the immediate postop. Alter this, no changes were found in the radiographical measurements. Conclusions. The use of an acetabular ring in revision THR enables the bringing down of the hip's center of rotation as well as an early restoration of both the structure of the area an the bone stock without any serious complications (AU)

Osteointegración de los vástagos no cementados en las artroplastias de cadera. ¿Son necesarias las revisiones clínicas anuales? / Osteointegration of uncemented stems in hip arthroplasty. Is annual clinical follow-up necessary?

Objetivo. Conocer los resultados de un vástago no cementado de cadera con exclusivo apoyo metafisario y un seguimiento mínimo de 5 años. Material y método. Estudio prospectivo de 338 vástagos del modelo Bihapro® implantados entre los años 1993 y 1998 para evaluar los cambios radiográficos durante este período. El vástago utilizado es anatómico con dos zonas, una metafisaria de superficie porosa e hidroxiapatita y otra distal lisa y afilada para evitar el contacto con el hueso cortical. Se estudiaron los criterios radiográficos de osteointegración a los 6 y 60 meses de la cirugía. Resultados. La serie final está formada por 316 vástagos. Los puentes óseos o puntos de soldadura fueron visibles en el 77,2% a los 6 meses y en el 83,9% a los 5 años. Las líneas radiotransparentes se detectaron en el 5,1% a los 6 meses y en el 4,7% a los 60 meses de seguimiento. La atrofia del calcar era evidente en el 18% a los 6 meses y en el 25% a los 60 meses. Las diferencias fueron estadísticamente significativas. Conclusiones. Los vástagos femorales no cementados de las artroplastias de cadera con apoyo metafisario y recubrimiento poroso e hidroxiapatita muestran signos radiográficos de integración ósea a los 6 meses que aumentan a los 60 meses, por lo que en los pacientes asintomáticos no es necesario realizar controles radiográficos anuales

Aim. To examine the outcome of uncemented hip stems with metaphyseal support alone and a minimum follow-up of 5 years. Materials and methods. Prospective study of 338 BiHAPro™ stems implanted between 1993 and 1998 to evaluate radiographic changes in this period. The stem is anatomically shaped with two sections: a metaphyseal section with a porous surface and hydroxyapatite coating and a tapered polished distal section designed to prevent cortical contact. Radiographic criteria were assessed at 6 and 60 months of surgery. Results. The final series consisted of 316 stems. Bone bridges or fusion points were visible in 77.2% at 6 months and in 83.9% at 5 years. Radiolucent lines were seen in 5.1% at 6 months and in 47% at 60 months of follow-up. Calcar atrophy was evident in 18% at 6 months and in 25% at 60 months. Differences were statistically significant. Conclusions: Uncemented femoral stems with metaphyseal support and a porous surface and hydroxyapatite coating show radiographic signs of osteointegration at 6 months that increase at 60 months, so it is not necessary to make annual radiographic revisions in asymptomatic patients

Extensively hydroxyapatite-coated stems in revision hip arthroplasty: A radiological mid-term follow-up study.

Femoral revision surgery involves the placement of a stable stem and the regeneration of the damaged bone structure. We retrospectively reviewed 36 cases of femoral revision surgery in which an extensively hydroxyapatite-coated porous stem was implanted with a five-year minimum follow-up. Nine of the cases had type I defects, 12 had type II defects, six had III-A defects and nine had III-B defects. The mean follow-up was 7.7 years (range 5-11). No cases of revision surgery, subsidence or displacement were detected. In 35 cases the integration of the stem had been achieved, the bone structure had been restored and there was no fibrous interface visible between the implant and the bone. The fully hydroxyapatite-coated stems thus proved a valuable alternative for femoral revision surgery in mild and moderate bone defects. (Hip International 2005; 15: 38-45).

Características, patrón de manejo y pronósticodel cáncer colorrectal / Characteristics, pattern of handling and prognosis of the colorectal cancer

Objetivos: investigar en una población de pacientes de nuestro ámbito clínico con cáncer colorrectal la distribución de sus edades, posibles diferencias en cuanto a los síntomas de presentación, características clínico-biológicas de sus tumores, diferencias en el tratamiento y en el pronóstico. Emplazamiento: Hospital de Jove de Gijón y el Hospital de referencia, Central de Asturias de Oviedo durante el periodo 1975-1999.Material y métodos: se realizó un estudio retrospectivo entre los meses de julio de 2001 a enero de 2002 que incluyo 473 pacientes del ámbito urbano y rural, diagnosticados de cáncer colorrectal. En todos se realizó el diagnóstico histológico de adenocarcinoma colorrectal. Se investigaron las características clásicas de los pacientes y de sus tumores así como su contenido de ADN, fase S, tipo de tratamiento y pronóstico de la enfermedad. Resultados: la edad media de los pacientes fue de 67,5 años (intervalo: 25-90 años). La década de mayor presentación fue la de los 70 años (33,2 por ciento) seguida de los 60 (32,6 por ciento) y la incidencia más baja correspondió a los <40 años (2,1 por ciento). La forma de presentación clínica que se evidenció con más frecuencia correspondió a la rectorragia (60,4 por ciento), se guida de la alteración del ritmo intestinal (47,9 por ciento) y el dolor (46,9 por ciento). En relación con las edades de los pacientes, encontramos diferencia significativa en cuanto al dolor (p<0,005), rectorragia, alteración del ritmo intestinal, tenesmo y adelgazamiento (p<0,0001). Existió una asociación estadística mente significativa entre la edad y la localización del tumor, siendo la localización derecha más frecuente en los pacientes de edad más avanzada (p<0,034). Asimismo, existieron diferencias significativas en cuanto al tratamiento en función de la edad de los pacientes, los de edad más avanzada recibieron menos frecuentemente tratamiento complementario (p<0,0001). Conclusiones: el presente estudio confirma que el cáncer colorrectal se presenta predominantemente en edades más avanzadas, que en los pacientes ancianos existe una predisposición de localización tumoral en el colon derecho, que no existe diferencia significativa de otras características tumorales ni del pronóstico de la enfermedad en función de la edad de los pacientes (AU)

[Expression and clinical significance of collagenase-3 (MMP-13) in gastric cancer]. / Expresión y significación clínica de la MMP-13 (colagenasa-3) en el cáncer gástrico.

BACKGROUND: MMP-13 (collagenase-3) is a metalloproteinase with potent degradative activity against a variety of elements of the extracellular matrix. Its expression has been described in some human carcinomas, where it seems to play a role in tumor progression and metastasis. The objective of this study was to investigate the expression and clinical significance of MMP-13 in gastric carcinomas. PATIENTS AND METHOD: MMP-13 expression was analyzed by immunohistochemistry in resected specimens from 44 patients with gastric adenocarcinoma. The mean ( standard error) follow-up period was 21.4 3.2 months. RESULTS: A total of 14 gastric carcinomas (31.8%) showed positive immunostaining for MMP-13. The percentage of MMP-13-positive tumors was significantly (p = 0.009) higher in stage IV carcinomas (69.2%) than in lower stages (I: 22.2%; II: 12.5%; and III: 14.3%), as well as in nonresectable tumors (R1 and R2) (61.5%) than in resectable carcinomas (R0) (19.4%) (p = 0.017). Likewise, MMP-13 tumor expression was significantly associated with shortened overall survival in both the entire group of patients (p = 0.0006) and in the subgroup of patients with resectable tumors (p = 0.018). CONCLUSIONS: Our results suggest that, in patients harboring gastric adenocarcinoma, MMP-13 tumor expression is associated with higher tumor aggressiveness and a poor prognosis.

Cytosolic levels of an estrogen-induced breast cancer-associated peptide (TFF1/pS2) in colorectal cancer: clinical significance and relationship with steroid receptors.

BACKGROUND: The Trefoil Factor 1 (TFF1/pS2), a peptide consisting of 60 amino acids, is the most abundant estrogen-induced messenger RNA in MCF-7 breast cancer cells and is also expressed by colorectal carcinomas. The objective of this work was to evaluate the cytosolic TFF1 content in colorectal carcinomas, its possible relationship with estrogen and progesterone receptors as well as with clinicopathological tumor parameters, and its potential prognostic significance. METHODS: Cytosolic TFF1 levels were examined by immunoradiometric assay in 178 patients with resectable colorectal cancer. The mean follow-up period was 32 months. RESULTS: There was a wide variability of cytosolic TFF1 levels in tumor-surrounding mucosa samples (0.09-42.5 ng/mg protein) as well as in tumors (0.01-270 ng/mg protein). Comparison of paired mucosa and carcinoma samples showed significantly higher TFF1 levels in tumors (mean: 17.1 ng/mg protein) than in mucosa samples (10 ng/mg protein) (p = 0.027). TFF1 levels were significantly higher in mucosa samples surrounding distal colon and rectal tumors (p = 0.0001) and in tumor samples obtained from older patients (p = 0.007). However, there were no significant differences in tumor TFF1 levels with respect to clinicopathological parameters such as the patient's sex, tumor location, stage, histological grade, ploidy, S-phase, or tumor estrogen and progesterone receptors. In addition, there was no significant relationship between tumor TFF1 levels and disease outcome. CONCLUSIONS: TFF1 may play an as yet undetermined role in the tumorigenesis of colorectal carcinomas. However, cytosolic levels of TFF1 do not seem to have any prognostic significance in colorectal carcinomas.

Importancia de la edad como factor pronóstico determinante de la variabilidad de las características clínico-patológicas de las pacientes con cáncer de mama / Importance of the age as factor determining prognosis of the variability of the clinical-pathological characteristics of patients with breast cancer

Objetivo: analizar la importancia de la edad como factor pronóstico en el cáncer de mama, lo cual podría tener importantes implicaciones en el tratamiento y manejo clínico de las pacientes. Material y métodos: estudio retrospectivo de 769 pacientes intervenidas quirúrgicamente por cáncer de mama (1983-1999). El tiempo medio de seguimiento fue de 34,4 meses. Se realizaron estudios bioquímicos para la determinación de receptores de estrógenos y progesterona, contenido de ADN y fase S. Resultados: la edad media de las 769 pacientes fue de 59 años, y la década de mayor presentación del cáncer de mama fue la de los 50 años (26,5 per cent). Encontramos una asociación estadísticamente significativa entre la edad de las pacientes y el tamaño tumoral (p<0,0001), grado histológico (p<0,018), contenido de receptores de estrógenos (p<0,014) y fase S (p<0,019). Las pacientes de menor edad tuvieron un mayor número de casos de tumores de menor ta maño, histológicamente menos diferenciados y con receptores estrogénicos negativos. El análisis univariante demostró que el mayor tamaño tumoral, el estadio nodal positivo y el grado histológico indiferenciado de los tumores estuvieron significativa y positivamente asociados con un menor tiempo libre de enfermedad y supervivencia total de las pacientes. Sin embargo, no existieron diferencias significativas en el pronóstico entre los diferentes grupos de pacientes clasificadas en función de sus distintas edades. Conclusiones: el presente estudio demuestra diferencias en las características clínicas, morfológicas y biológicas de los carcinomas mamarios en función de la edad de las pacientes. Sin embargo , nuestros resultados no demostraron diferencias significativas en el pronóstico de las pacientes en función de su edad. Ello podría tener importantes implicaciones de cara al tratamiento y manejo clínico de las pacientes, principalmente de aquellas de más avanzada edad (AU)