Construction of a recombinant adenovirus co-expressing truncated human prostate-specific membrane antigen and mouse 4-1BBL genes and its effect on dendritic cells

Our aim was to construct a recombinant adenovirus co-expressing truncated human prostate-specific membrane antigen (tPSMA) and mouse 4-1BBL genes and to determine its effect on dendritic cells (DCs) generated from bone marrow suspensions harvested from C57BL/6 mice for which the effect of 4-1BBL on DCs is not clear, especially during DCs processing tumor-associated antigen. Replication deficient adenovirus AdMaxTM Expression System was used to construct recombinant adenovirus Ad-tPSMA-internal ribosome entry site-mouse 4-1BBL (Ad-tPSMA-IRES-m4-1BBL) and Ad-enhanced green fluorescent protein. Day 7 proliferating DC aggregates generated from C57BL/6 mice were collected as immature DCs and further mature DCs were obtained by lipopolysaccharide activated immature DCs. After DCs were exposed to the recombinant adenovirus with 250 multiplicity of infection, the expression of tPSMA and m4-1BBL proteins were detected by Western blot, and the apoptosis and phenotype of DCs were analyzed by flow cytometry. Cytokines (IL-6 and IL-12) in the supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Proliferation of T cells was detected by allogeneic mixed lymphocyte reactions. The tPSMA and m4-1BBL proteins were expressed correctly. The apoptosis rate of DCs transfected with Ad-tPSMA-IRES-m4-1BBL was 14.6 percent, lower than that of control DCs. The expression of co-stimulatory molecules [CD80 (81.6 ± 5.4 percent) and CD86 (80.13 ± 2.81 percent)] up-regulated in Ad-tPSMA-IRES-m4-1BBL-pulsed DCs, and the level of IL-6 (3960.2 ± 50.54 pg/mL) and IL-12 (249.57 ± 12.51 pg/mL) production in Ad-tPSMA-IRES-m4-1BBL-transduced DCs were significantly higher (P < 0.05) than those in control DCs. Ad-tPSMA-IRES-m4-1BBL induced higher T-cell proliferation (OD450 = 0.614 ± 0.018), indicating that this recombinant adenovirus can effectively enhance the activity of DCs.

Refining the assessment of the sensitivity and specificity of diagnostic tests, with applications to prostate cancer screening and non-small cell lung cancer staging

Los cálculos del rendimiento de las pruebas diagnósticas suelen presentarse en tablas de 2x2 con filas horizontales para los resultados positivos y negativos obtenidos con la prueba evaluada, y con columnas verticales para los resultados positivos y negativos obtenidos con el patrón de oro. Esta manera de presentar los datos visualmente, así como las sensibilidades y especificidades basadas en ella, le imprimen carácter binario a la prueba y al patrón de oro. Pero cuando los resultados de la prueba pertenecen a una de varias categorías ordenadas, a menudo se utilizan curvas de las características funcionales de la prueba (o curvas ROC, por receiver operator characteristic curve) para indicar que esta no es binaria. Tratar el patrón de oro como si fuese binario también es problemático porque implica que toda enfermedad se comporta uniformemente, con el resultado de que a todos los casos se les trata como si fuesen intercambiables. No obstante, hay ciertos tumores, por ejemplo, que exigen más tratamiento que otros y que por lo tanto también exigen mayor detección. En el presente trabajo proponemos el uso de una tabla refinada que clasifica a los tumores en función de lo que se sabe de su susceptibilidad al tratamiento, con lo cual se pretende lograr una evaluación más informativa de las pruebas que la proporcionada por la tabla de 2x2. A manera de ejemplo presentamos una tabla de 2x3 en la cual se refina la medición del antígeno específico de la próstata (AEP) teniendo en cuenta el resultado de la palpación rectal. Dicho resultado se usa como indicador de la necesidad de tratar los cánceres prostáticos que se detectan o que no se detectan mediante la prueba del AEP. Un segundo ejemplo aplica los mismos conceptos a la tomografía por emisión de positrones y a la tomografía computadorizada cuando se usan para la estadificación del cáncer pulmonar no microcítico. Se usaría más información si se adoptara la estructura de 2x3 para configurar la tabla.

Molecular detection of prostate specific antigen in patients with prostate cancer or benign prostae hyperplasia the first investigation from Iran

Prostate cancer is the second common form of cancer in men. Detection of circulating Prostate Specific Antigen [PSA] transcripts has effectively been used for early diagnosis of prostate cancer cells. This investigation employed a reverse transcriptase polymerase chain reaction [RT-PCR] technique to distinguish the patients with either localized or metastatic prostate cancer [CaP] vs. Benign Prostate Hyperplasia [BPH] and control subjects, as compared with clinical and pathological records. With reservation of ethical issues, blood samples were collected from 60 cases. Based on pathological and clinical findings, 25 patients [20 with localized cancer, 5 with metastatic], 22 with BPH, and 13 healthy [including 3 females] subjects as negative controls, were selected from Shariati, Mehrad, Sina,, Khatam and Atie Hospitals in Tehran, Iran. RT-PCR for a 260 bp PSA transcript was then performed. Clinical and pathological records were used for the assessment and comparison of PSA RT-PCR results. None of the control subjects and BPH [with 7 exceptions] were found positive by RT-PCR [Relative specificity= 72.7%]. In patients with prostate cancer, 21 out of 25 were found PSA positive [Relative sensitivity=83.4%] and the remaining 3 have been shown to be PSA negative [Positive predictive value= 83.4%]. All of 5 metastatic patients [100%] revealed PSA positive results. Our data reflects the clinical relevance and significance of RT-PCR results as assessed with clinical and pathological examinations. PSA RT-PCR might be used as a powerful means for diagnosis, even when either pathological or clinical findings are negative, and could be employed for further molecular epidemiology surveys

O PSA como marcador tumoral é confiável? / PSA as tumor marker is reliable?

Considerado como um dos melhores marcadores tumorais, o antígeno prostático específico (PSA) é largamente utilizado. Valores acima de 4,ng/ml podem significar câncer de próstata (CaP). No entanto, doenças benignas podem alterar o seu valor e o câncer de próstata pode apresentar-se sem alterá-lo. O exame digital da próstata (EDP) e a dosagem de PSA são as melhores formas de diagnosticar e acompanhar os pacientes com mais de 40 anos.

One of the best tumors markers ever known, the prostate-specific antigen (PSA) is well advantage today. Values above 4.ng/ml can demonstrate prostate cancer. Although benign diseases would change the values of PSA and prostate cancers would not. Digital prostate exam (DPE) and PSA are the best way to diagnostic and watch men with over 40 years.

Morphological Feature correlation with serum tumour markers in prostatic carcinoma

To find out Gleason grades, scores and to see the correlation of these morphological features with tumour markers in prostatic carcinoma. Design: A descriptive study. Place and Duration of Study: The study was conducted at the Departments of Histopathology and Chemical Pathology, Armed Forces Institute of Pathology, Rawalpindi, over a period of one year. Materials and Fifty cases of prostatic carcinoma were studied. Gleason grades and score of tumour were determined by doing haematoxylin and eosin [H and E] staining. Pre-operative serum prostatic acid phosphatase [PAP] and prostate-specific antigen [PSA] assays were carried out in these cases. The patients seen were between 50-102 years of age with an average of 70.9 years. There were 49 cases of adenocarcinoma and 01 case of mixed adeno and transitional cell carcinoma of prostate. Twenty-eight [56%] patients had Gleason score of 5-7. Twenty-nine [58%] patients were having serum PSA levels between 10.0 ng/ml and 50.0 ng/ml. Thirteen [26%] cases showed PSA assays > 50 ng/ml. The sensitivity of PSA test was 84% in these cases. Thirty-five [70%] patients were having PAP values > 3.7 U/l [sensitivity 70%]. The Gleason grading system is a specific morphological predictor. The serum PSA showed better sensitivity and specificity with Gleason grades and scores as compared to serum PAP. The serum PAP levels showed better correlation with morphological features as compared to serum PSA

Autoanticuerpos contra antígeno específico en hiperplasia benigna y cáncer de próstata / Autoantibodies againts prostatic specific antigen in benign hyperplasia and prostate cancer

Por medio de ELISA (Enzyme linked immunosorbent assay), se llevó a cabo la determinación de autoanticuerpos contra el antígeno específico de próstata en los sueros de dos grupos de pacientes: uno con hiperplasia prostática benigna y otro con cáncer de próstata. Los resultados encontrados confirman la presencia de autoanticuerpos en los sueros de ambos grupos, sin embargo, no se demostró diferencia estadísticamente significativa en la concentración de autoanticuerpos contra el antígeno específico de próstata