Adipose Tissue Hypoxia Correlates with Adipokine Hypomethylation and Vascular Dysfunction.

Obesity is characterized by the accumulation of dysfunctional adipose tissues, which predisposes to cardiometabolic diseases. Our previous in vitro studies demonstrated a role of hypoxia in inducing adipokine hypomethylation in adipocytes. We sought to examine this mechanism in visceral adipose tissues (VATs) from obese individuals and its correlation with cardiometabolic risk factors. We propose an involvement of the hypoxia-inducible factor, HIF1α, and the DNA hydroxymethylase, TET1. Blood samples and VAT biopsies were obtained from obese and non-obese subjects (n = 60 each) having bariatric and elective surgeries, respectively. The analyses of VAT showed lower vascularity, and higher levels of HIF1α and TET1 proteins in the obese subjects than controls. Global hypomethylation and hydroxymethylation were observed in VAT from obese subjects along with promoter hypomethylation of several pro-inflammatory adipokines. TET1 protein was enriched near the promotor of the hypomethylated adipokines. The average levels of adipokine methylation correlated positively with vascularity and arteriolar vasoreactivity and negatively with protein levels of HIF1α and TET1 in corresponding VAT samples, serum and tissue inflammatory markers, and other cardiometabolic risk factors. These findings suggest a role for adipose tissue hypoxia in causing epigenetic alterations, which could explain the increased production of adipocytokines and ultimately, vascular dysfunction in obesity.

Prediction of immune surveillance responsive metastatic prostate cancer in pelvic lymph node and emergence of surveillance unresponsive/resistant metastatic cells.

BACKGROUND: Immune cells (lymphocytes and macrophages) provide the microenvironment for immune surveillance of metastatic prostate cancer (PCa) cells in pelvic lymph nodes. We have hypothesized that degeneration and/or apoptosis of metastatic PCa cells in pelvic lymph nodes can distinguish between aggressive and non-aggressive metastatic disease in patients. Our objective was to define the relationship between metastatic cell lysis and the presence of immune cells. MATERIALS AND METHODS: We studied archival primary PCa (n=38) and cancer-positive regional pelvic nodes (n=32) from the same patients undergoing radical retropubic prostatectomy at the Minneapolis Veterans Affairs Medical Center. RESULTS: Using morphological and immunohistochemical features of immune and metastatic cancer cells, we have identified progression of metastasis in the nodal compartments. Nodal parenchyma contained small, intermediate and large metastatic nodules/tumors. Immune surveillance occurred primarily in small tumors and surveillance was either absent or greatly reduced in intermediate and large tumors in nodes. Metastatic nodules/cells were lysed or became apoptotic when under immune-surveillance, as indicated by pyknotic nuclei and cytoplasm, the latter still had remnants of prostate specific antigen (PSA) staining. Metastatic cells without surveillance did not exhibit morphological features of cell degeneration (lysis) or apoptosis. Metastatic cells under immune-surveillance had an inverse relationship with those without immune-surveillance. This relationship differed from node to node and patient to patient. CONCLUSION: We have shown that at least two populations of metastatic cells were present in the nodes; the first group of cells was under immune surveillance, as indicated by limited to wide-spread cell lysis/apoptosis, and the second group did not exhibit morphological evidence of cell lysis indicating emergence of surveillance-unresponsive (resistant) metastatic cells. These criteria can be used to distinguish metastatic cancer that is expected to be responsive to immunotherapy from that which would show little or no benefit from such treatment. Enhancement of immune surveillance and other treatments can be used to treat surveillance-unresponsive (resistant) disease to improve survival of patients.

Cathepsin B expression is similar in African-American and Caucasian prostate cancer patients.

BACKGROUND: Increased incidence and mortality of prostate cancer (PCa) suggest that U.S. African-American men have more invasive cancer than Caucasian men. Invasive PCa requires several proteases, including the cysteine protease cathepsin B (CB), for degradation of basement membrane and extracellular matrix proteins prior to cancer cell migration across biological compartments. Our objective was to determine whether CB immunostaining patterns, in relation to clinical data, could distinguish invasive PCa in African-American and Caucasian patients. PATIENTS AND METHODS: Fifty Gleason score 6/7 PCa cases were selected for similar clinical data with benign prostatic hyperplasia (BPH) samples as controls. Immunostainings were imaged directly from microscope slides to a computer using a digital camera. Data were quantified using Metamorph software, analyzed using the two-sample t-test and confirmed by multiple regression. RESULTS: Ratios of CB to its endogenous inhibitor stefin A (SA) immunostainings were greater in PCa than BPH, but were not significantly different in PCa of either race. The African-American patients did not show increased CB immunostaining, indicating that the contribution of this protease to invasiveness was similar in both races. CONCLUSION: When veterans received equal medical care at the Minneapolis Veterans Affairs Medical Center, African-American patients did not show increased PCa invasiveness. Our conclusion is supported by analysis of post-surgery serum total PSA levels and cancer cell invasion to margins/capsules, seminal vesicles and/or lymph nodes. Invasiveness of PCa does not appear to be race-dependent. The previous conclusion of race-based differences in PCa requires re-evaluation with respect to the role of proteases (such as CB, matrix metalloproteinase) in invasion and metastasis of cancer cells.

Prediction of pelvic lymph node metastasis by the ratio of cathepsin B to stefin A in patients with prostate carcinoma.

BACKGROUND: Pathologic grade and/or histologic score, extraprostatic extension indicated by invasion of the prostatic capsule, margin, and/or seminal vesicles by prostate cancer cells, serum total prostate-specific antigen (PSA), free PSA, complexed PSA levels and/or their ratios, regional pelvic lymph node metastases, and clinical staging have been used to diagnose and monitor the treatment of prostate carcinoma (PC) patients. The Gleason grading system is also used to grade/score a patient's stage of disease, with lower to higher scores indicating progression of PC. However, Gleason's system cannot be used to distinguish biologically aggressive PCs within a single Gleason score. Our objective was to identify subpopulations (or clones) of aggressive prostate cancers within an individual Gleason score by utilizing biological molecule(s) that also facilitate cancer cell invasion to prostatic stroma and metastasis to the lymph nodes. MATERIALS AND METHODS: Specimens were collected from 97 patients with PC and from 8 patients with benign prostatic hyperplasia. These patients had not been treated with hormonal and/or chemotherapeutic agents before undergoing a prostatectomy at the Minneapolis Veterans Affairs Medical Center. Formalin-fixed, paraffin or paraplast-embedded prostate tissue sections were stained with hematoxylin and eosin for pathologic diagnosis and adjacent sections were stained for for immunohistochemical study. We also collected data on age, race, extraprostatic extension, margin status, seminal vesicle, and lymph node invasion by cancer cells, clinical stage at prostatectomy, and mortality/survival data, including the available presurgery and postsurgery serum total PSA and prostatic acid phosphatase concentrations in patients. Immunohistochemical localization of mouse or rabbit anti-cathepsin B (CB) antibody IgG and mouse antihuman stefin (cystatin) A IgG was quantified using a computer-based image analysis system equipped with Metamorph software. RESULTS: CB and stefin A identified aggressive and less aggressive clones of PCs within an individual Gleason score. Tumors with a Gleason Score of 6 that are similar histologically and morphologically were heterogeneous with respect to the ratios of CB to stefin A (CB > stefin A, CB = stefin A, and CB < stefin A). We also found a significant positive association (P = 0.0066) between ratios of CB and stefin A (CB > stefin A) and the incidence of pelvic lymph node metastases, but not with ratios of CB less than stefin A and/or ratios of CB equal to stefin A. Patients with Gleason 7 PCs had a higher incidence of positive lymph nodes than those with Gleason Score 6 tumors. Our data indicated that mortality rates increased in patients when the ratios of CB were greater than stefin A. CONCLUSIONS: PC within an individual Gleason score is a heterogeneous tumor that contains clones or subpopulations of aggressive and less aggressive tumors that can be defined by the ratios of CB to stefin A. PC with an aggressive clone can be identified when the ratio of CB is greater than that of stefin A. Less aggressive clones are identified when the ratio of CB is less than that of stefin A or when the ratio of CB is equal to that of stefin A. The ratios of CB to stefin A can be used in the differential diagnosis and treatment of patients with PC. This is the first report to identify phenotypes of aggressive and less aggressive PCs within a Gleason score.

Routine bladder biopsy after bacille Calmette-Guérin treatment: is it necessary?

OBJECTIVES: To evaluate the need for routine biopsy at first follow-up after intravesical bacille Calmette-Guérin (BCG) therapy for superficial transitional cell cancer of the bladder (TCCB). We examined the potential role of voided urine cytology and cystoscopy as screening tools to determine which patients should undergo biopsy. METHODS: The records of 71 patients with TCCB who received a total of 95 courses of BCG were reviewed. The pathology findings before BCG and the cystoscopic, voided cytologic, and biopsy results after BCG were recorded. RESULTS: In 54 cases, the cystoscopic and/or cytologic findings were abnormal. In 19 (35%) of these 54 cases, TCCB was found on biopsy after BCG. In 41 cases, both cystoscopy and cytology were normal. In 1 of these 41 cases, a small superficial low-grade TCCB was found on biopsy. An abnormality on voided cytology or cystoscopy was more likely to occur after treatment for carcinoma in situ. CONCLUSIONS: Patients with papillary TCCB who have negative cystoscopic and negative urine cytologic results can safely be spared routine transurethral bladder biopsy with its associated cost and morbidity. However, patients with carcinoma in situ are very likely to have persistent abnormal cytologic or abnormal cystoscopic findings warranting investigation with biopsy and may benefit from routine scheduled biopsy.

Enterocolite necrosante em neonatos a termo, lactentes e pré-escolares / Necrotizing enterocolítis in term neonates, older children and preschools

Os autores analisam 300 autopsias em que foram identificados 71 casos de enterocolite necrosante em recém-nascidos a termo e pré-escolares. Diarréia e desnutriçäo foram fatores predisponentes comumente associados. Dois tipos histopatológicos distintos da doença foram demonstrados: enterocolite necrosante difusa, cujo padräo é semelhante ao descrito em recé-nascidos pré-termo, e enterocolite necrosante pápulo-necrótica, padräo menos freqüente que o difuso, e que se apresenta principalmente como lesöes crônicas isoladas em crianças maiores. Peritonite purulenta ou fibrinosa ocorreu mais comumente por contigüidade à lesäo intestinal, sendo identificados apenas quatro casos de perfuraçäo. Constatou-se que, em analogia ao recém-nascido pré-termo com asfixia perinatal, cuja suscetivilidade a infecçöes é maior devido à imaturidade do seu sistema imunológico, diarréia crônica e desnutriçäo em recém-nascidos a termo e crianças maiores causam deficiência imunológica e alteraçäo na barreira intestinal, que associadas a hipovolemia (por desidrataçäo) e sépsis levam a isquemia e invasäo da mucosa intestinal pela flora micobacteriana local produzindo, freqüentemente, um padräo histológico distinto daquele descrito para enterocolite necrosante em recém-nascidos pré-termo de risco. Os autores chamem atençäo para a desnutriçäo como fator predisponente para o desenvolvimento da doença