Ki67 expression at Kasai portoenterostomy as a prognostic factor in patients with biliary atresia.

BACKGROUND: Biliary atresia is a rare paediatric biliary obliteration disease with unknown aetiology, and is the most common indication for paediatric liver transplantation (LT). However, no consensus for predicting Kasai portoenterostomy (KP) outcomes using liver histological findings exists. Ki67 is a popular biomarker for measuring and monitoring cellular proliferation. METHODS: Ki67 (clone, MIB-1) liver parenchyma expression was measured by immunohistochemical staining of samples from living donors and patients with biliary atresia to assess its value in predicting outcomes after KP. RESULTS: Of 35 children with biliary atresia, 13 were native liver survivors (NLS), 17 were non-NLS, and five had primary LT. The median proportion of Ki67 immunostained areas in donors and patients with biliary atresia at KP was 0·06 and 0·99 per cent respectively. Univariable analysis identified a high proportion of Ki67 areas, high Ki67 cell numbers and high Ki67-positive/leucocyte common antigen-positive cell numbers at KP as significant predictors of poor native liver survival after KP (hazard ratio 9·29, 3·37 and 12·17 respectively). The proportion of Ki67 areas in the non-NLS group was significantly higher than that in the NLS group (1·29 versus 0·72 per cent respectively; P = 0·001), and then decreased at LT (0·32 per cent versus 1·29 per cent at KP; P < 0·001). CONCLUSION: This study has demonstrated the clinical data and time course of Ki67 expression in patients with biliary atresia. High Ki67 expression at KP may be an important predictor of native liver survival following the procedure.

ANTECEDENTES: La atresia biliar (biliary atresia, BA) es una enfermedad pediátrica rara que consiste en una obstrucción biliar de etiología desconocida, y es la indicación pediátrica más frecuente de trasplante hepático (liver transplantation, LT). Sin embargo, no existe consenso para predecir los resultados de la portoenterostomía de Kasai (Kasai portoenterostomy, KP) en base a los hallazgos histológicos hepáticos. El Ki67 es un biomarcador conocido para medir y controlar la proliferación celular. MÉTODOS: Se midieron los niveles de expresión del parénquima hepático de Ki67 (clon, MIB-1) por tinción inmunohistoquímica de las muestras de cinco donantes vivos y 35 pacientes con BA, para evaluar su valor predictivo de los resultados de la KP. RESULTADOS: Los pacientes con BA incluían 13 sobrevivientes con hígado nativo (native liver survivors, NLS), 17 no NLS y 5 pacientes que se sometieron inicialmente a LT. La proporción media de las áreas de expresión de Ki67 en donantes y pacientes con BA en KP fue de 0,06% y 0,99%, respectivamente. El análisis univariado identificó una alta proporción de áreas de Ki67, un alto número de células Ki67, un alto número de células Ki67 positivas (+)/leucocitos (LCA/CD45) + en KP como predictores significativos de una peor supervivencia del hígado nativo después de KP (cociente de riesgos instantáneos, hazard ratio, HR 9,29, 3,37 y 12,17, respectivamente). La proporción de las áreas Ki67 fueron significativamente superiores en los pacientes sin NLS que en los pacientes con NLS (P = 0,001). Entre los pacientes sin hígado nativo, los niveles de Ki67 disminuyeron posteriormente de acuerdo con la presencia de una lesión hepática irreparable, tales como son los hígados con BA en LT (en KP versus en LT = 1,29% versus 0.32%; P < 0,001). CONCLUSIÓN: Demostramos los datos clínicos y la evolución temporal de la expresión de Ki67 en los pacientes con BA. El alto nivel de expresión de Ki67 en KP puede ser un predictor importante para la supervivencia del hígado nativo después de KP.

The Warburg effect and tumour immune microenvironment in extramammary Paget's disease: overexpression of lactate dehydrogenase A correlates with immune resistance.

BACKGROUND: Extramammary Paget's disease (EMPD) is a rare malignant skin cancer. One of the hallmarks of cancers, including EMPD, is an enhancement of aerobic glycolysis, which is also known as the Warburg effect. In the last step of glycolysis, the enzyme lactate dehydrogenase A (LDHA) catalyzes the conversion of pyruvate to lactic acid, the accumulation of which contributes to the creation of an acidic tumour microenvironment. This in turn results in immunosuppression in various types of cancers. However, the contribution of these pathways has not been well-studied in EMPD. OBJECTIVE: To investigate the significance of the Warburg effect and its contribution to the tumour immune microenvironment in EMPD. METHODS: The mRNA expression levels of molecules involved in glycolysis and immune-related cytokines were examined by ddPCR. The number of immune cells was assessed by immunohistochemistry (IHC). RESULTS: The levels of two glycolytic enzymes, HK2 and LDHA, in tumour tissues were significantly increased compared to those in paired-normal tissues. IHC analyses revealed increased numbers of PD-L1+ , PD-1+ , CD163+ M2 macrophages, Iba1+ macrophages and Foxp3+ Tregs that were associated with high LDHA levels in EMPD. ddPCR demonstrated that multiple cytokines including IL-4, IL-6, IL-10, TGF-ß and CCL-2 were upregulated and associated with high LDHA levels in EMPD. Statistical analyses showed that IL-6 mRNA expression correlated with the number of CD163+ , Iba-1+ and Foxp3+ cells. CONCLUSION: The Warburg effect contributes to immunomodulation in the tumour microenvironment and further elucidation may lead to better understanding of the pathogenesis of EMPD.

Morphological lymphocytic reaction, patient prognosis and PD-1 expression after surgical resection for oesophageal cancer.

BACKGROUND: Immune checkpoint inhibitors, such as antibody against programmed cell death protein (PD-1), have demonstrated antitumour effects in patients with malignancies, including oesophageal cancer. A lymphocytic reaction observed by pathological examination is a manifestation of the host immune response to tumour cells. It was hypothesized that a stronger lymphocytic reaction to tumours might be associated with favourable prognosis in oesophageal cancer. METHODS: Using a database of resected oesophageal cancers, four morphological components of lymphocytic reactions (peritumoral, intranest, lymphoid and stromal) to tumours were evaluated in relation to clinical outcome, PD-1 expression by immunohistochemistry and total lymphocyte count in blood. RESULTS: Resected oesophageal cancer specimens from 436 patients were included in the study. Among the four morphological components, only peritumoral reaction was associated with patient prognosis (multivariable P for trend <0·001); patients with a higher peritumoral reaction had significantly longer overall survival than those with a lower reaction (multivariable hazard ratio 0·48, 95 per cent c.i. 0·34 to 0·67). The prognostic effect of peritumoral reaction was not significantly modified by other clinical variables (all P for interaction >0·050). Peritumoral reaction was associated with total lymphocyte count in the blood (P < 0·001), supporting the relationship between local immune response and systemic immune competence. In addition, higher morphological peritumoral reaction was associated with high PD-1 expression on lymphocytes in tumours (P = 0·034). CONCLUSION: These findings should help to improve risk-adapted therapeutic strategies and help stratify patients in the future clinical setting of immunotherapy for oesophageal cancer.

ANTECEDENTES: Los inhibidores de los puntos de control inmunitario (checkpoints) (p.ej. los anticuerpos anti-PD-1) han demostrado efectos antitumorales en pacientes con tumores malignos, incluido el cáncer de esófago. La reacción linfocítica detectada en estudios anatomopatológicos es una manifestación de la respuesta inmune del huésped a las células tumorales. Se estableció la hipótesis de que una mayor reacción linfocítica a los tumores podría asociarse con un mejor pronóstico en el cáncer de esófago. MÉTODOS: Usando una base de datos de 436 cánceres de esófago resecados, se evaluaron cuatro componentes morfológicos (peritumoral, intra-epitelial, linfoide y estromal) de las reacciones linfocíticas a tumores en relación con los resultados clínicos, la expresión inmunohistoquímica de PD-1 y el recuento total de linfocitos en sangre. RESULTADOS: De los cuatro componentes, solamente la reacción peritumoral se asoció con el pronóstico del paciente (P multivariable para tendencia < 0,001): los pacientes con mayor reacción peritumoral presentaron una supervivencia global significativamente más prolongada que aquellos pacientes con menor reacción peritumoral (cociente de riesgos instantáneos multivariable, hazard ratio, HR: 0,48; i.c. del 95%: 0,34 -0,67; P <0,001). El efecto pronóstico de la reacción peritumoral no se modificó significativamente por otras variables clínicas (todas las P para la interacción > 0,05). La reacción peritumoral se asoció con el recuento total de linfocitos en la sangre (P < 0,001), lo que respalda la relación entre la respuesta inmune local y la competencia inmune sistémica. Además, una elevada reacción morfológica peritumoral se asoció con una alta expresión de PD-1 en linfocitos tumorales (P = 0,034). CONCLUSIÓN: Estos hallazgos deberían ayudar a mejorar las estrategias terapéuticas adaptadas al riesgo y contribuir a estratificar a los pacientes en el entorno clínico futuro de la inmunoterapia para los pacientes con cáncer de esófago.

Possible involvement of the M2 anti-inflammatory macrophage phenotype in growth of human gliomas.

Within tumours, many non-neoplastic cells such as fibroblasts, endothelial cells, and macrophages assist tumour growth by producing various growth factors and pro-angiogenic cytokines. Various tumour-derived molecules drive tumour-associated macrophages towards an anti-inflammatory phenotype (M2) and thus promoting tumour growth. Here we investigated microglia/macrophage differentiation in glioma tissues by means of immunostaining of paraffin-embedded glioma samples. The number of microglia/macrophages with positive staining for CD163 and CD204, which are believed to be markers for M2 macrophages, was correlated with the histological grade of the gliomas. The ratio of M2 macrophages in the tumour-associated microglia/macrophages was also associated with the histological grade. Culture supernatant from the glioma cell line can stimulate macrophages to develop into the M2 phenotype in vitro. Macrophage colony-stimulating factor (M-CSF), which strongly induces M2 polarization of macrophages, was significantly correlated with histological malignancy and with the proportion of M2 microglia/macrophages in vivo. In addition, the proportion of M2 microglia/macrophages and M-CSF expression in tumour cells correlated well with proliferation of glioblastoma cells. These results suggest that tumour-derived M-CSF induces a shift of microglia/macrophages towards the M2 phenotype, which influences tumour growth. Evaluation of the proportion of M2 microglia/macrophages and M-CSF expression in tumour tissue would be useful for assessment of microglia/macrophage proliferative activity and the prognosis of patients with gliomas.

Identification of Lck-derived peptides applicable to anti-cancer vaccine for patients with human leukocyte antigen-A3 supertype alleles.

The identification of peptide vaccine candidates to date has been focused on human leukocyte antigen (HLA)-A2 and -A24 alleles. In this study, we attempted to identify cytotoxic T lymphocyte (CTL)-directed Lck-derived peptides applicable to HLA-A11(+), -A31(+), or -A33(+) cancer patients, because these HLA-A alleles share binding motifs, designated HLA-A3 supertype alleles, and because the Lck is preferentially expressed in metastatic cancer. Twenty-one Lck-derived peptides were prepared based on the binding motif to the HLA-A3 supertype alleles. They were first screened for their recognisability by immunoglobulin G (IgG) in the plasma of prostate cancer patients, and the selected candidates were subsequently tested for their potential to induce peptide-specific CTLs from peripheral blood mononuclear cells of HLA-A3 supertype(+) cancer patients. As a result, four Lck peptides were frequently recognised by IgGs, and three of them - Lck(90-99), Lck(449-458), and Lck(450-458) - efficiently induced peptide-specific and cancer-reactive CTLs. Their cytotoxicity towards cancer cells was mainly ascribed to HLA class I-restricted and peptide-specific CD8(+) T cells. These results indicate that these three Lck peptides are applicable to HLA-A3 supertype(+) cancer patients, especially those with metastasis. This information could facilitate the development of peptide-based anti-cancer vaccine for patients with alleles other than HLA-A2 and -A24.

Expression of APOBEC3G in kidney cells.

The apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G (APOBEC3G), a member of the APOBEC family possessing DNA mutator activity through cytosine deamination, is reported to play an important role in host defense against infections such as those of hepatitis B virus and human immunodeficiency virus. Here, we examined the expression of APOBEC3G in human kidney cells to better understand its biological role against infection. APOBEC3G was immunohistochemically detectable in kidney mesangial cells and also to some extent in kidney epithelial tubular cells. In addition, overexpression of APOBEC3G was shown in renal carcinoma tissues and cell lines. APOBEC3G expression was upregulated by inflammatory cytokines, such as interferon, interleukin-6, and tumor necrosis factor. These results may provide new insight into the role of APOBEC3G in host defense against viral infection and cancer.

Abdominal helical CT: evaluation of optimal doses of intravenous contrast material--a prospective randomized study.

PURPOSE: To determine the optimal dose of intravenous contrast material for helical computed tomography (CT) of the abdomen on the basis of patient weight. MATERIALS AND METHODS: A prospective randomized study of helical CT of the abdomen was performed by using different doses of intravenous contrast material in 221 patients (mean body weight, 57.3 kg) who were assigned randomly to three groups receiving 1.5, 2.0, or 2.5 mL/kg or a fixed dose of 100 mL of iopamidol 300. The degree of enhancement was scored visually. The CT numbers (HU) of the aorta, portal vein, liver, and pancreas were obtained at three levels of the abdomen. RESULTS: In arterial enhancement, the 2.0 mL/kg, 2.5 mL/kg, and fixed-dose groups were significantly better than the 1.5 mL/kg group, but there was no significant difference among the 2.0 mL/kg, 2.5 mL/kg, or fixed-dose groups. The degree of enhancement of the liver, pancreas, and portal vein increased with larger doses. At visual analysis, hepatic parenchymal enhancement was graded as good or excellent in 64% of the 1.5 mL/kg, 85% of the 2.0 mL/kg, 94% of the 2.5 mL/kg, and 65% of the fixed-dose groups. CONCLUSION: When dose was tailored to patient weight, the use of 2.0-2.5 mL/kg of intravenous contrast material produced better results than did 1.5 mL/kg or a fixed dose. Arterial enhancement did not differ among the 2.0 mL/kg, 2.5 mL/kg, or fixed-dose groups.

MR imaging of Tornwaldt's cysts.

OBJECTIVE: The purpose of this study was to assess the characteristics of Tornwaldt's cysts as revealed by routine MR studies. MATERIALS AND METHODS: We retrospectively reviewed MR images of the brain in 1208 consecutive subjects who ranged in age from 3 weeks to 93 years (mean, 57.1 years). The signal intensity, shape, and size of Tornwaldt's cysts were assessed. Patients with Tornwaldt's cysts were then questioned about the presence of persistent nasal discharge, occipital headaches, and halitosis and an unpleasant taste in the mouth and about a history of adenoidectomy. RESULTS: Tornwaldt's cysts were found in 23 patients (1.9%) who ranged in age from 39 to 78 years (mean, 57.3 years). Of the 23 Tornwaldt's cysts, all were isointense to CSF on T2-weighted images and hyperintense to gray matter on the fluid-attenuated inversion-recovery images. The cysts showed high signal intensity compared with muscle on T1-weighted images. Nineteen cysts were round and four were oval. The mean size of the lesions was 6.0 mm in the major axis and 5.5 mm in the minor axis. Two patients with Tornwaldt's cysts had persistent nasal discharge and occipital headaches, and another patient had occipital headaches alone. None of the patients had undergone an adenoidectomy. CONCLUSION: Lesions consistent with Tornwaldt's cysts were found in 1.9% of the routine MR studies of the brain. The cysts had high signal intensity on T1-weighted, T2-weighted, and fluid-attenuated inversion-recovery images.

CT evaluation of gastric lesions with three-dimensional display and interactive virtual endoscopy: comparison with conventional barium study and endoscopy.

OBJECTIVE: This study was undertaken to assess the feasibility of three-dimensional (3D) CT rendering using shaded-surface display (SSD) and ray sum display and virtual endoscopic images of the stomach for simultaneous evaluation of intraluminal and extraluminal abnormalities compared with conventional upper gastrointestinal barium studies and endoscopy. SUBJECTS AND METHODS: Our prospective study consisted of 39 patients with gastric lesions (17 gastric carcinomas, nine gastric polyps, five gastric varices, five gastric submucosal tumors, one lymphoma, one case of Menetrier's disease, and one gastric erosion) detected by endoscopy and barium study. All 3D CT images were reconstructed using SSD, ray sum display, and virtual endoscopic techniques. Three-dimensional images were evaluated for ability to reveal the range and morphologic features of the gastric lesions. RESULTS: All SSD, ray sum display, and virtual endoscopic images successfully revealed five of the eight early-stage gastric carcinomas and all nine advanced-stage gastric carcinomas. Submucosal tumors were revealed on 3D CT approximately as well as on conventional endoscopy. Interactive evaluation of virtual endoscopic images and multiplanar reconstructions provided useful information regarding intraluminal and submucosal gastric involvement by gastric varices, submucosal tumor, advanced gastric carcinomas, and lymphoma. This kind of information could not be obtained by conventional endoscopy or double-contrast study. CONCLUSION: Three-dimensional CT used in conjunction with virtual CT endoscopy proved helpful in identifying gastric lesions. Also, virtual CT endoscopic images with the interactive display of multiplanar reconstructions proved useful in identifying both intraluminal and submucosal components.

Usefulness of diffusion-weighted MRI with echo-planar technique in the evaluation of cellularity in gliomas.

The purpose of this study was to evaluate the utility of diffusion-weighted magnetic resonance imaging (MRI) with echo-planar imaging (EPI) technique in depicting the tumor cellularity and grading of gliomas. Twenty consecutive patients (13 men and 7 women, ranging in age from 13 to 69 years) with histologically proven gliomas were examined using a 1.5 T superconducting imager. Tumor cellularity, analyzed with National Institutes of Health Image 1.60 software on a Macintosh computer, was compared with the minimum apparent diffusion coefficient (ADC) and the signal intensity on the T2-weighted images. The relationship of the minimum ADC to the tumor grade was also evaluated. Tumor cellularity correlated well with the minimum ADC value of the gliomas (P = 0.007), but not with the signal intensity on the T2-weighted images. The minimum ADC of the high-grade gliomas was significantly higher than that of the low-grade gliomas. Diffusion-weighted MRI with EPI is a useful technique for assessing the tumor cellularity and grading of gliomas. This information is not obtained with conventional MRI and is useful for the diagnosis and characterization of gliomas.

CT angiography in vascular intervention for steno-occlusive diseases: role of multiplanar reconstruction and source images.

The aim of this study was to assess the efficacy of CT angiography for steno-occlusive diseases before and after interventional procedures, focusing on the role of multiplanar reconstruction (MPR) and source images. 17 patients with 20 steno-occlusive lesions underwent CT angiography before and after interventional procedures. For each lesion, the percentage stenosis obtained on CT angiography was compared with that on conventional angiography. In addition, MPR and source images were evaluated for the presence of wall thickening and calcification before interventional procedures, and the presence of dissection and luminal shape after interventional procedures. These findings were compared with those of conventional angiography. Although the percentage stenosis depicted on CT angiography correlated well with that on conventional angiography, MPR and source images clearly demonstrated the effect of intervention and the residual stenosis. MPR and source images clearly depicted wall thickening, wall calcification, the presence of dissection and the luminal shape. CT angiography provides useful information before intervention, while MPR and source images are of value in evaluating arterial wall abnormalities and morphological changes associated with interventional procedures.

MR of epidermoids with a variety of pulse sequences.

PURPOSE: To assess the usefulness of fluid-attenuated inversion recovery (FLAIR) and constructive interference in steady state (CISS) sequences in depicting epidermoid tumors. METHODS: Six patients with surgically confirmed epidermoid tumors in the subarachnoid space were examined with T1-weighted MR imaging with a spin-echo sequence, and with T2- and proton density-weighted imaging with a fast spin-echo sequence, a FLAIR sequence, and a CISS sequence. In the qualitative analysis, three observers compared the five sequences for visibility of tumors and presence of artifacts. A quantitative analysis was also performed by measuring the contrast-to-noise ratio. RESULTS: On visual assessment, the FLAIR sequence depicted all tumors as hyperintense relative to cerebrospinal fluid. The CISS sequence depicted all tumors as hypointense relative to cerebrospinal fluid and was considered to show tumor extension better than the FLAIR sequence. At quantitative analysis, the mean contrast-to-noise ratios of tumor to cerebrospinal fluid on T1-, T2-, and proton density-weighted images, and on FLAIR and CISS sequences were 2.85, 3.41, 4.42, 16.13, and 20.23, respectively. The contrast-to-noise ratios for the FLAIR and CISS sequences were significantly higher than those for the T1-, T2-, and proton density-weighted sequences. The contrast-to-noise ratio was not significantly different between FLAIR and CISS sequences, although the CISS sequence was slightly superior. CONCLUSION: CISS and FLAIR sequences depicted epidermoid tumors in the subarachnoid spaces better than conventional spin-echo images did. The CISS sequence produced a relatively constant contrast between the tumors and less artifactual interference.

T2 shortening in the visual cortex: effect of aging and cerebrovascular disease.

PURPOSE: To evaluate the effect of aging and cerebrovascular disease on T2 shortening in the visual cortex at MR imaging. METHODS: MR images of 72 neurologically normal subjects (45 men and 27 women, 35 to 92 years old) and 32 (13 men and 19 women, 54 to 92 years old) with cerebrovascular disease were evaluated retrospectively. On T2-weighted spin-echo images, the signal intensity of the visual, motor, and sensory cortices was divided into three grades and compared with the signal intensity of the frontal subcortical white matter. RESULTS: Decreased signal intensity (grade III) was rarely seen in the visual and sensory cortices of the neurologically normal subjects who were less than 60 years old. The signal intensity of the motor cortex decreased rapidly after the age of 50 years. At 61 to 70 years of age, 53% of these subjects had grade III intensity, and at age 71 years or older, 94% had reached grade III. The frequency of progression from grade I to grade III was lower in the visual cortex than in the motor cortex; 22% of these subjects had grade III appearance at age 61 to 70 years, and at age 71 years older, 56% had reached grade III. In patients with cerebrovascular disease who were older than 60 years of age, the frequency of grade III signal intensity in the visual cortex was almost equal to that in the neurologically normal subjects. CONCLUSIONS: T2 shortening in the visual cortex is frequently seen in neurologically normal older persons. These findings are compatible with a previously reported histochemical study of normal iron deposition in the visual cortex. Cerebrovascular disease has no effect on T2 shortening in the visual cortex.