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.

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).

[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.

Evaluation of blood pressure control after bilateral glomectomy: effects of propranolol treatment.

The effects of bilateral glomectomy on arterial blood pressure and heart rate and the response to the pressor tests were studied in a women of 37. Arterial pressure and ECG were recorded by the continuous unrestricted Oxford method. Hypotension, increased blood pressure variability and tachycardia rapidly developed 2 days after surgery, indicating sympathetic hyperactivity and/or deactivation of vagal tone. Wide blood pressure fluctuations were present on standing and abnormal blood pressure responses to head up tilting and to the Valsalva maneuver were observed reflecting a less prompt and precise control of blood pressure. Treatment with propranolol reduced tachycardia and blood pressure variability. After 5 and 17 months of this therapy propranolol was withheld for 36 h. Tachycardia was still present and blood pressure fluctuations were recorded on assuming the upright posture. However, the heart rate and pressure response to the Valsalva maneuver were normal after 17 months, indicating an improvement of the reflex control of blood pressure.