The Co-Use of Stromal Vascular Fraction and Bone Marrow Concentrate for Tendon Healing.

OBJECTIVE: The Achilles tendon is the most frequently injured tendon in the human body, despite being the strongest. Many conventional treatments including medication, surgical interventions, and physical therapy are available, however, the desired results are often not achieved. Stromal vascular fraction (SVF) and bone marrow concentrate (BMC) are two additional cellular treatment options. The purpose of this study is to evaluate the effect of SVF and BMC, used as a combination, for the treatment of Achilles tendon injuries. METHODS: Five male New Zealand rabbits were used for each of the 6 study groups. A 3-mm of SVF and BMC were injected on the Achilles tendons at certain ratios. The histological results were classified by the Movin grading system for tendon healing. The collagen type-I and type-III structures in the tendons were examined by immunohistochemical evaluation. The expressions of tendon-specific genes were also examined by using the RT-PCR method to analyze tendon healing. RESULTS: Histological and immunohistochemical evaluation indicated that tendons receiving the SVF and BMAC mixture performed better than control and individual groups (p < 0.05). Moreover, RT-PCR evaluation showed that mixture-receiving groups were the closest similar to the uninjured group (p < 0.05). CONCLUSION: The combined use of BMC and SVF improved Achilles tendon healing when compared to the individual use of each mixture.

Idiopathic granulomatous hypophysitis mimicking adenoma.

Hypophysitis is a rare disease of pituitary gland, which, although it is usually a primary lesion, can also occur secondary to systemic conditions. Granulomatous hypophysitis is an inflammatory disease condition which accounts for less than 1% of all cellular lesions and can mimic adenoma. A 32-year-old woman presented with weight gain, galactorrhea and blurred vision. The MRI showed a cystic, nodular lesion in the intermediate lobe of the pituitary gland and the initial diagnosis was adenoma. She underwent surgery and the histopathology revealed granulomas composed of epithelioid histiocytes, multinuclear giant cells and mononuclear inflammatory cells. Inflammatory diseases of the pituitary gland are much less frequent than pituitary adenomas and idiopathic granulomatous hypophysitis is extremely rare. Histopathology and the ruling out of a systemic cause are the gold standards for its diagnosis.

Idiopathic granulomatous hypophysitis mimicking adenoma

Hypophysitis is a rare disease of pituitary gland, which, although it is usually a primary lesion, can also occur secondary to systemic conditions. Granulomatous hypophysitis is an inflammatory disease condition which accounts for less than 1% of all cellular lesions and can mimic adenoma. A 32-year-old woman presented with weight gain, galactorrhea and blurred vision. The MRI showed a cystic, nodular lesion in the intermediate lobe of the pituitary gland and the initial diagnosis was adenoma. She underwent surgery and the histopathology revealed granulomas composed of epithelioid histiocytes, multinuclear giant cells and mononuclear inflammatory cells.Inflammatory diseases of the pituitary gland are much less frequent than pituitary adenomas and idiopathic granulomatous hypophysitis is extremely rare. Histopathology and the ruling out of a systemic cause are the gold standards for its diagnosis.(AU)

La hipofisitis es una enfermedad rara de la glándula pituitaria, y a pesar de ser fundamentalmente una enfermedad primaria, puede ser también secundaria a enfermedades sistémicas. La hipofisitis granulomatosa es una enfermedad inflamatoria que representa menos del 1% de todas las lesiones celulares, y remedar al adenoma. Mujer de 32 años de edad ingresada en el hospital con aumento de peso, galactorrea y visión borrosa. La RM reflejó una lesión quística y nodular en el lóbulo intermedio de la glándula pituitaria, que fue operada con diagnóstico primario de adenoma. El examen microscópico reveló granulomas formados por histiocitos epitelioides, células gigantes multinucleares y células inflamatorias mononucleares. Las enfermedades inflamatorias de la glándula pituitaria son muy raras en comparación con los adenomas pituitarios. La hipofisitis granulomatosa idiopática es una de ellas. El criterio de referencia para este diagnóstico es realizar un examen histopatológico, y descartar una causa sistémica.(AU)

Can chlorogenic acid reduce oxidative stress and in an experimental spinal cord injury?

BACKGROUND: We aimed to investigate antioxidant and neuroprotective properties of chlorogenic acid in spinal cord injury (SCI). METHODS: Twenty-one rats were divided into three groups. Laminectomy was performed in group L (n=7), spinal cord trauma was induced in group T (n=7), and spinal cord trauma was induced and chlorogenic acid treatment was started in group C (n=7). Blood samples were collected to analyze baseline values and the 12th h, 1st day, 3rd day, and 5th day catalase, native thiol (NT), total thiol (TT), disulfide (SS), SS/TT, SS/NT, and NT/TT levels. Functional analysis with Basso-Beattie and Bresnahan scores was performed at the same time points. Total antioxidant status (TAS), total oxidative stress, oxidative stress index, and cyclooxygenase-2 (Cox-2) were examined in the spinal cord of rats euthanized on day 7; results were statistically analyzed. RESULTS: On day 7, catalase levels in Group C were significantly higher than baseline levels, whereas those in Group T were significantly lower than baseline levels; Group L showed no significant difference (p=0.008). SS values on day 7 were lower in Group T than in Groups C and L. Group C showed the lowest decrease in NT/TT level after trauma. On day 7, SS/TT level was high in Group T but stable in Groups C and L (p=0.04). Histopathological examination revealed significantly lower Cox-2 and TAS levels in Group C than in Group T (p=0.003, p=0.017, respectively). CONCLUSION: In this study, SCI was primarily examined through thiol-SS balance, and it was demonstrated by experimental models that chlorogenic acid has antioxidant and neuroprotective effects in SCI.

Diagnostic approach in TFE3-rearranged renal cell carcinoma: a multi-institutional international survey.

Transcription factor E3-rearranged renal cell carcinoma (TFE3-RCC) has heterogenous morphologic and immunohistochemical (IHC) features.131 pathologists with genitourinary expertise were invited in an online survey containing 23 questions assessing their experience on TFE3-RCC diagnostic work-up.Fifty (38%) participants completed the survey. 46 of 50 participants reported multiple patterns, most commonly papillary pattern (almost always 9/46, 19.5%; frequently 29/46, 63%). Large epithelioid cells with abundant cytoplasm were the most encountered cytologic feature, with either clear (almost always 10/50, 20%; frequently 34/50, 68%) or eosinophilic (almost always 4/49, 8%; frequently 28/49, 57%) cytology. Strong (3+) or diffuse (>75% of tumour cells) nuclear TFE3 IHC expression was considered diagnostic by 13/46 (28%) and 12/47 (26%) participants, respectively. Main TFE3 IHC issues were the low specificity (16/42, 38%), unreliable staining performance (15/42, 36%) and background staining (12/42, 29%). Most preferred IHC assays other than TFE3, cathepsin K and pancytokeratin were melan A (44/50, 88%), HMB45 (43/50, 86%), carbonic anhydrase IX (41/50, 82%) and CK7 (32/50, 64%). Cut-off for positive TFE3 fluorescent in situ hybridisation (FISH) was preferably 10% (9/50, 18%), although significant variation in cut-off values was present. 23/48 (48%) participants required TFE3 FISH testing to confirm TFE3-RCC regardless of the histomorphologic and IHC assessment. 28/50 (56%) participants would request additional molecular studies other than FISH assay in selected cases, whereas 3/50 participants use additional molecular cases in all cases when TFE3-RCC is in the differential.Optimal diagnostic approach on TFE3-RCC is impacted by IHC and/or FISH assay preferences as well as their conflicting interpretation methods.

Robot Assisted Radical Cystectomy Outcomes in Micropapillary and Plasmacytoid Variants.

PURPOSE: To compare the patients who underwent robot assisted radical cystectomy (RARC) and extended pelvic lymph node dissection (EPLND) and whose pathology result was reported as micropapillary variant (MV), plasmacytoid variant (PV) and pure urothelial carcinoma (PUC). MATERIALS AND METHODS: The data of 133 patients who underwent RARC and EPLND with the postoperative pathology results reported as MV, PV and PUC were analyzed. According to the postoperative pathology results, patients were divided into two groups in initial analyses as variant pathologies group (n=14) and PUC group (n=119). In secondary analyses, patients were divided into three groups as MV group (n=7), PV group (n=7) and PUC group (n=119). The operative data, oncologic outcomes and complications were compared between the groups. RESULTS: Median operation time and estimated blood loss were significantly increased in variant pathologies group (P <0.001 and P = .001, respectively). The postoperative pathological T stage, positive surgical margin rate and lymph node involvement were also significantly increased in variant pathologies (P = .001, P = 0.004, P <0.001, respectively). Kaplan-Meier analysis revealed significant decrease in OS and CSS times in PV group compared to PUC group (P = .048 and P = .016, respectively). CONCLUSION: MV and PV are rarely seen variant pathologies with higher pathological T stages. RARC is a minimally invasive surgical technique that can be performed successfully by an experienced surgical team with low morbidity rates and similar oncological results, even in challenging cases.

Multicenter Multireader Evaluation of an Artificial Intelligence-Based Attention Mapping System for the Detection of Prostate Cancer With Multiparametric MRI.

OBJECTIVE. The purpose of this study was to evaluate in a multicenter dataset the performance of an artificial intelligence (AI) detection system with attention mapping compared with multiparametric MRI (mpMRI) interpretation in the detection of prostate cancer. MATERIALS AND METHODS. MRI examinations from five institutions were included in this study and were evaluated by nine readers. In the first round, readers evaluated mpMRI studies using the Prostate Imaging Reporting and Data System version 2. After 4 weeks, images were again presented to readers along with the AI-based detection system output. Readers accepted or rejected lesions within four AI-generated attention map boxes. Additional lesions outside of boxes were excluded from detection and categorization. The performances of readers using the mpMRI-only and AI-assisted approaches were compared. RESULTS. The study population included 152 case patients and 84 control patients with 274 pathologically proven cancer lesions. The lesion-based AUC was 74.9% for MRI and 77.5% for AI with no significant difference (p = 0.095). The sensitivity for overall detection of cancer lesions was higher for AI than for mpMRI but did not reach statistical significance (57.4% vs 53.6%, p = 0.073). However, for transition zone lesions, sensitivity was higher for AI than for MRI (61.8% vs 50.8%, p = 0.001). Reading time was longer for AI than for MRI (4.66 vs 4.03 minutes, p < 0.001). There was moderate interreader agreement for AI and MRI with no significant difference (58.7% vs 58.5%, p = 0.966). CONCLUSION. Overall sensitivity was only minimally improved by use of the AI system. Significant improvement was achieved, however, in the detection of transition zone lesions with use of the AI system at the cost of a mean of 40 seconds of additional reading time.

Use of Aspirin and Statins in Relation to Inflammation in Benign Prostate Tissue in the Placebo Arm of the Prostate Cancer Prevention Trial.

Aspirin and statin use may lower the risk of advanced/fatal prostate cancer, possibly by reducing intraprostatic inflammation. To test this hypothesis, we investigated the association of aspirin and statin use with the presence and extent of intraprostatic inflammation, and the abundance of specific immune cell types, in benign prostate tissue from a subset of men from the placebo arm of the Prostate Cancer Prevention Trial. Men were classified as aspirin or statin users if they reported use at baseline or during the 7-year trial. Presence and extent of inflammation were assessed, and markers of specific immune cell types (CD4, CD8, FoxP3, CD68, and c-KIT) were scored, in slides from end-of-study prostate biopsies taken irrespective of clinical indication, per trial protocol. Logistic regression was used to estimate associations between medication use and inflammation measures, adjusted for potential confounders. Of 357 men included, 61% reported aspirin use and 32% reported statin use. Prevalence and extent of inflammation were not associated with medication use. However, aspirin users were more likely to have low FoxP3, a T regulatory cell marker [OR, 5.60; 95% confidence interval (CI), 1.16-27.07], and statin users were more likely to have low CD68, a macrophage marker (OR, 1.63; 95% CI, 0.81-3.27). If confirmed, these results suggest that these medications may alter the immune milieu of the prostate, which could potentially mediate effects of these medications on advanced/fatal prostate cancer risk.

FDG Hepatic Superscan Due to Metastatic Infiltration of Prostate Cancer.

A 69-year-old man with prostate cancer presented to the hospital with 2 weeks' history of fever, abdominal distension, and fatigue. Laboratory findings showed signs of acute liver failure, and marked elevation of lactate dehydrogenase and tumor marker levels. Abdominal CT showed hepatomegaly with multiple hypodense lesions in both lobes, suggesting metastases. FDG PET/CT scan shows hypermetabolism unusually in the liver with significantly suppressed heart and brain activity, reminiscent of an FDG hepatic superscan. The hypermetabolic lesions confirmed with Tru-Cut needle biopsy of the liver as metastasis of prostate cancer.

Autologous stem cell-derived chondrocyte implantation with bio-targeted microspheres for the treatment of osteochondral defects.

BACKGROUND: Chondral injury is a common problem around the world. Currently, there are several treatment strategies for these types of injuries. The possible complications and problems associated with conventional techniques lead us to investigate a minimally invasive and biotechnological alternative treatment. Combining tissue-engineering and microencapsulation technologies provide new direction for the development of biotechnological solutions. The aim of this study is to develop a minimal invasive tissue-engineering approach, using bio-targeted microspheres including autologous cells, for the treatment of the cartilage lesions. METHOD: In this study, a total of 28 sheeps of Akkaraman breed were randomly assigned to one of the following groups: control (group 1), microfracture (group 2), scaffold (group 3), and microsphere (group 4). Microspheres and scaffold group animals underwent adipose tissue collection prior to the treatment surgery. Mesenchymal cells collected from adipose tissue were differentiated into chondrocytes and encapsulated with scaffolds and microspheres. Osteochondral damage was conducted in the right knee joint of the sheep to create an animal model and all animals treated according to study groups. RESULTS: Both macroscopic and radiologic examination showed that groups 3 and 4 have resulted better compared to the control and microfracture groups. Moreover, histologic assessments indicate hyaline-like cartilage formations in groups 3 and 4. CONCLUSION: In conclusion, we believe that the bio-targeted microspheres can be a more effective, easier, and safer approach for cartilage tissue engineering compared to previous alternatives.

Diagnosing atypia of undetermined significance in thyroid fine needle aspiration samples using nuclear scoring.

BACKGROUND: Atypia of undetermined significance (AUS) is an indeterminate category in the Bethesda system for reporting thyroid cytopathology. Cytological features described as atypia are not always observed in every case, and it is difficult to determine how the small population of cells with enlarged nuclei, a few grooves, and rare elongated nuclei should be classified. Therefore, there is inter-intra observer variability considering these cell types, even though the cytological criteria are well defined. Therefore, this study aimed to establish a nuclear scoring system to help in the differential diagnosis of AUS. METHODS: Fine needle aspiration (FNA) samples that showed AUS and had surgical follow-up were included in this study. The aspirate was scored for the presence of intanuclear cytoplasmic inclusions, nuclear grooves, overlapping, enlargement, and elongation individually. The total nuclear score for each case was calculated. Statistical analysis of the association between each nuclear feature and the presence of papillary thyroid cancer (PTC) in the surgical specimens was performed. Cut-off points from the total score of these nuclear features were also calculated. RESULTS: Nuclear grooves and overlapping were more common in malignant cases (p < 0.001 and p = 0.048, respectively). A cut-off point of ≥5.5 for the total score was sensitive and specific for defining malignancy. CONCLUSION: The risk of PTC was higher in nodules with more prominent nuclear overlapping or nuclear groove in their FNA samples. In order to achieve a more confident AUS diagnosis, our scoring system can be helpful for thyroid FNA samples.

Prognostic significance of the programmed death ligand 1 expression in clear cell renal cell carcinoma and correlation with the tumor microenvironment and hypoxia-inducible factor expression.

BACKGROUND: Clear cell renal cell carcinoma (ccRCC) is the most common renal malignancy. Hypoxia-inducible factors, HIF-1α and HIF-2α, are expressed in the majority of ccRCC. Targeting immune checkpoints with the blockade of PD-1 and its ligand PD-L1 reorganizes T-cell activity in tumor microenvironment and provides important antitumor responses. PD-L1 upregulation has been found to be hypoxia-inducible factor (HIF) dependent. Our aim is to demonstrate the association between PD-L1 and HIF expression and to reveal the role of PD-L1 in prognosis and its association with tumor microenvironment. METHODS: Surgical specimens from 145 patients diagnosed with ccRCC, who had undergone radical or partial nephrectomy, were retrospectively analyzed. Immunohistochemistry on tissue microarrays (TMA) was performed to demonstrate expressions of PD-L1, HIF-1α, and HIF-2α in tumor cells and PD-1, CD4, and CD8 in lymphocytes to assess lymphocyte density in tumor microenvironment. RESULTS: PD-L1 tumor cell expression was detected in 20/125 (13.8%) cases, which correlated with higher levels of PD-1, CD4, CD8 and HIF-2α expression. Low or high expression of HIF-1α was similar in PD-L1-positive cases. When PD-L1-positive cases were compared with negative ones, there was no significant difference in terms of prognostic factors. However, the number of WHO/ISUP grade 3-4 tumors was significantly higher in PD-L1-positive cases than in negative ones. CONCLUSION: PD-L1 tumor cell expression is strongly associated with increased HIF-2α expression and presence of dense lymphocytic infiltration in ccRCCs. Our findings confirm that PD-L1 positivity is associated with high ISUP nucleolar grade. The association between PD-L1, HIF, and lymphocyte density in tumor microenvironment must be clarified and especially taken into account in combination treatment.

A Comparative Study of Receptor-Targeted Magnetosome and HSA-Coated Iron Oxide Nanoparticles as MRI Contrast-Enhancing Agent in Animal Cancer Model.

Magnetosomes are specialized organelles arranged in intracellular chains in magnetotactic bacteria. The superparamagnetic property of these magnetite crystals provides potential applications as contrast-enhancing agents for magnetic resonance imaging. In this study, we compared two different nanoparticles that are bacterial magnetosome and HSA-coated iron oxide nanoparticles for targeting breast cancer. Both magnetosomes and HSA-coated iron oxide nanoparticles were chemically conjugated to fluorescent-labeled anti-EGFR antibodies. Antibody-conjugated nanoparticles were able to bind the MDA-MB-231 cell line, as assessed by flow cytometry. To compare the cytotoxic effect of nanoparticles, MTT assay was used, and according to the results, HSA-coated iron oxide nanoparticles were less cytotoxic to breast cancer cells than magnetosomes. Magnetosomes were bound with higher rate to breast cancer cells than HSA-coated iron oxide nanoparticles. While 250 µg/ml of magnetosomes was bound 92 ± 0.2%, 250 µg/ml of HSA-coated iron oxide nanoparticles was bound with a rate of 65 ± 5%. In vivo efficiencies of these nanoparticles on breast cancer generated in nude mice were assessed by MRI imaging. Anti-EGFR-modified nanoparticles provide higher resolution images than unmodified nanoparticles. Also, magnetosome with anti-EGFR produced darker image of the tumor tissue in T2-weighted MRI than HSA-coated iron oxide nanoparticles with anti-EGFR. In vivo MR imaging in a mouse breast cancer model shows effective intratumoral distribution of both nanoparticles in the tumor tissue. However, magnetosome demonstrated higher distribution than HSA-coated iron oxide nanoparticles according to fluorescence microscopy evaluation. According to the results of in vitro and in vivo study results, magnetosomes are promising for targeting and therapy applications of the breast cancer cells.

Active nano/microbilayer hemostatic agents for diabetic rat bleeding model.

Patients with diabetes mellitus have an increased cardiovascular risk due to the abnormality of hemostatic system components. Therefore, hemostasis is an important concept when considering that diabetics are under risk due to potential bleeding complications during surgical operation. The aim of our study was to examine the efficiency of a fabricated nano/microbilayer hemostatic dressing for bleeding control in diabetic patients. For this purpose, we prepared a nano/microbilayer hemostatic dressing that has a porous sublayer, including chitosan (CTS), bacterial cellulose (BC) as basement and active agents in coagulation cascade, such as vitamin K (Vit K), protamine sulfate (PS), and kaolin (Kao) as a filler and an upper layer consisting of silk fibroin (SF) or SF/phosphatidylcholine (PC) blend to achieve complete hemostasis in diabetic rats. Coagulative performances of the prepared hemostatic dressings were examined by the determination of bleeding time, blood loss, and mortality rate through diabetic rat femoral artery injury model. The percent of diabetic rat blood absorption was found to be 247 ± 5% for gauze as opposed to 2214 ± 56% for SF-coated PS/BC/CTS. Vit K-reinforced within 138 s and SF-coated BC/CTS hemostatic dressings within 144 s showed a rapid coagulation time. In vivo coagulation studies demonstrated that hemostatic agent-reinforced BC/CTS hemostatic dressing, especially PS/BC/CTS showed a significant hemostatic plug formation. This study suggests that the high positive charge and porosity give to these hemostatic agents reinforced hemostatic dressings the ability to rapidly swell and to promote the accumulation of red blood cells and platelets through electrostatic interactions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1573-1585, 2017.

Peripheral Zone Inflammation Is Not Strongly Associated With Lower Urinary Tract Symptom Incidence and Progression in the Placebo Arm of the Prostate Cancer Prevention Trial .

BACKGROUND: Intraprostatic inflammation has been associated with lower urinary tract symptom (LUTS) progression. However, prior studies used tissue removed for clinical indications, potentially skewing inflammation extent or biasing the association. We, therefore, evaluated inflammation and LUTS incidence and progression in men who underwent biopsy of the prostate peripheral zone irrespective of indication. MATERIALS AND METHODS: We developed nested case-control sets in men in the placebo arm of the Prostate Cancer Prevention Trial who were free of clinical BPH and had a protocol-directed year 7 biopsy. Cases had baseline IPSS <15 and year 7 IPSS of 8-14 (low, N = 47), 15-19 (incident moderate, N = 42), or ≥20 (incident high, N = 44). Controls had baseline and year 7 IPSS <8 (N = 41). For progression from IPSS <8, cases had baseline to year 7 IPSS slope >75th percentile (N = 46) and controls had a slope <25th percentile (N = 45). For progression from IPSS = 8-14, cases had a slope >75th percentile (N = 46) and controls had a slope <25th percentile (N = 46). We reviewed three H&E-stained biopsy cores per man to determine prevalence of ≥1 core with inflammation and mean extent (%) of tissue area with inflammation. RESULTS: Inflammation prevalence in low cases (64%) was similar to controls (66%), but higher in moderate (69%) and high (73%) cases (P-trend = 0.4). Extent did not differ across LUTS categories (P-trend = 0.5). For progression from IPSS < 8, prevalence (65%, P = 0.9) and extent (2.5%, P = 0.8) in cases did not differ from controls (64%, 2.7%). For progression from IPSS 8-14, prevalence in cases (52%) was lower than in controls (78%, P = 0.009), while extent was higher in cases (5.3%) than controls (3.6%), especially in men with ≥1 core with inflammation (10.1% versus 4.6%, P = 0.06). CONCLUSION: Peripheral zone intraprostatic inflammation is not strongly associated with LUTS incidence or progression. Prostate 76:1399-1408, 2016. © 2016 Wiley Periodicals, Inc.

Molecular evidence that invasive adenocarcinoma can mimic prostatic intraepithelial neoplasia (PIN) and intraductal carcinoma through retrograde glandular colonization.

Prostate cancer often manifests as morphologically distinct tumour foci and is frequently found adjacent to presumed precursor lesions such as high-grade prostatic intraepithelial neoplasia (HGPIN). While there is some evidence to suggest that these lesions can be related and exist on a pathological and morphological continuum, the precise clonal and temporal relationships between precursor lesions and invasive cancers within individual tumours remain undefined. Here, we used molecular genetic, cytogenetic, and histological analyses to delineate clonal, temporal, and spatial relationships between HGPIN and cancer lesions with distinct morphological and molecular features. First, while confirming the previous finding that a substantial fraction of HGPIN lesions associated with ERG-positive cancers share rearrangements and overexpression of ERG, we found that a significant subset of such HGPIN glands exhibit only partial positivity for ERG. This suggests that such ERG-positive HGPIN cells either rapidly invade to form adenocarcinoma or represent cancer cells that have partially invaded the ductal and acinar space in a retrograde manner. To clarify these possibilities, we used ERG expression status and TMPRSS2-ERG genomic breakpoints as markers of clonality, and PTEN deletion status to track temporal evolution of clonally related lesions. We confirmed that morphologically distinct HGPIN and nearby invasive cancer lesions are clonally related. Further, we found that a significant fraction of ERG-positive, PTEN-negative HGPIN and intraductal carcinoma (IDC-P) lesions are most likely clonally derived from adjacent PTEN-negative adenocarcinomas, indicating that such PTEN-negative HGPIN and IDC-P lesions arise from, rather than give rise to, the nearby invasive adenocarcinoma. These data suggest that invasive adenocarcinoma can morphologically mimic HGPIN through retrograde colonization of benign glands with cancer cells. Similar clonal relationships were also seen for intraductal carcinoma adjacent to invasive adenocarcinoma. These findings represent a potentially undervalued indicator of pre-existing invasive prostate cancer and have significant implications for prostate cancer diagnosis and risk stratification.

Cytoplasmic PTEN protein loss distinguishes intraductal carcinoma of the prostate from high-grade prostatic intraepithelial neoplasia.

Intraductal carcinoma of the prostate is a marker of aggressive disease. However, intraductal carcinoma exists on a morphologic continuum with high-grade prostatic intraepithelial neoplasia (PIN) and distinguishing intraductal carcinoma from PIN is a common diagnostic dilemma with significant clinical implications. We evaluated whether immunostains for PTEN and ERG can sensitively identify intraductal carcinoma and accurately distinguish it from high-grade PIN. A combined immunostain for PTEN, ERG, p63 and CK903 was developed and validated. Radical prostatectomy specimens with lesions meeting criteria for intraductal carcinoma (n=45), intraductal cribriform proliferations falling short of intraductal carcinoma (n=15), and PIN lesions (n=39) were retrospectively identified and assessed for PTEN and ERG. Cytoplasmic PTEN loss was identified in 84% (38/45) of the intraductal carcinoma and 100% (15/15) of intraductal cribriform proliferation cases. In contrast, cytoplasmic PTEN loss was never observed in PIN (0/39; P<0.0001). Of the 53 cases of intraductal carcinoma or intraductal cribriform proliferation with cytoplasmic PTEN loss, it was homogeneously lost in 42 cases (79%). Weak, focal nuclear positivity for PTEN was retained in 31 of these 42 cases (74%). ERG expression was identified in 58% (26/45) of intraductal carcinoma and 67% (10/15) of intraductal cribriform proliferations compared with 13% (5/39) of PIN. Concordance between the PTEN/ERG status of the intraductal carcinoma lesions and the concurrent invasive carcinoma was high (>95% and P<0.0001 for each), and substantially less for PIN and the concurrent invasive tumor (83% for PTEN and 67% for ERG; P=NS for each). Cytoplasmic PTEN loss occurs in the majority of intraductal carcinoma and intraductal cribriform proliferation cases. Cytoplasmic PTEN loss was never observed in PIN (100% specificity). Our study identifies PTEN loss as a potentially useful marker to distinguish intraductal carcinoma from PIN and provides a plausible molecular explanation for why intraductal carcinoma is associated with poor prognosis.

Lymphatic and capillary invasion patterns in triple negative breast cancer.

Triple negative breast cancer (TNBC) comprises approximately 15 to 20 per cent of all breast cancer cases. Many studies have detected less lymph node metastasis in TNBC than sporadic breast cancers. In this study, we studied capillary and lymphatic invasion in tumors of patients with TNBC. To differentiate the capillary invasion and lymphovascular invasion. We used the Antihuman CD34 and antihuman D2-40 antibodies. Antihuman CD34 antibodies stain the blood vessels and lymphatics. However, antihuman D2-40 antibodies stain lymphatics specifically. Two experienced pathologists blinded to clinical data evaluated capillary and lymphatic invasion existence in 39 TNBC patients' tumor samples. Tumor samples were immunohistochemically stained with CD34 (endothelial cell marker) and D2-40 (podoplanin, a membrane protein, specific for lymphatic endothelium). The CD34-positive samples were categorized into two groups depending on their reaction with D2-40: lymphatic (D2-40-positive) and capillary (D2-40-negative) invasion. We have detected vascular invasion in 15 of 39 samples (38.5%) with CD34. Among those, capillary invasion was found in 14 (35.9%) and lymphatic invasion in three (7.7%) and both in two (5.1%) tumors. We did not find any significant correlation among capillary invasion, lymphatic invasion, vascular invasion, tumor grade, menopause status, history of cancer, and TNM. Capillary invasion is more commonly observed than lymphatic invasion in patients with TNBC. This finding supports the fact that more hematogenous metastasis (spreading) and less lymph node metastasis are seen in patients with TNBC.

EGFR expression and gene copy number in triple-negative breast carcinoma.

Most basal-like breast carcinomas are estrogen receptor negative, progesterone receptor negative, and cerb-B2/HER-2/neu negative--the so-called triple-negative breast carcinomas--with high epidermal growth factor receptor (EGFR) expression, which makes EGFR a target of treatment. We evaluated EGFR expression by immunohistochemistry (IHC) with two different clones (EGFR.31G7 and EGFR.25) and gene copy number by fluorescence in situ hybridization (FISH) with Locus specific identifier EGFR/CEP 7 dual probe in 62 triple-negative breast carcinomas. Any complete or incomplete membranous and/or cytoplasmic expression was regarded as IHC positive. Cases showing gene amplification (a ratio of EGFR gene to chromosome 7 of ≥ 2 or 15 copies per cell in ≥ 10% of cells) and high polysomy (≥ 4 copies in ≥ 40% of cells) were considered FISH po sitive. We detected EGFR.31G7 positivity in 38 of 62 cases (61.4%), which was composed of 12 of 62 (19.4%) cytoplasmic, 14 of 62 (22.6%) incomplete membranous, and 12 of 62 (19.4%) complete membranous staining. Among 38 of 49 (77.6%) EGFR.25-positive cases, 7 of 49 (14.3%) exhibited cytoplasmic, 10 of 49 (20.4%) exhibited incomplete membranous, and 21 of 49 (42.9%) exhibited complete membranous staining pattern. Ten of 62 (16.1%) FISH-positive cases were identified; 1 of 62 (1.6%) showed amplification, and the rest showed high polysomy. All FISH-positive cases were also found to be IHC positive (P = 0.01) by both EGFR clones. The amplified case displayed strong complete membranous staining with both clones. Among the high polysomic cases; 4 of 9 (44.4%) incomplete membranous, 4 of 9 (44.4%) complete membranous and 1 of 9 (11.1%) cytoplasmic expression of EGFR.31G7, and 6 of 8 (75%) complete membranous and 2 of 6 (25%) cytoplasmic expression of EGFR.25 were detected. Here, we report that membranous EGFR expression is associated with increased gene copy number (P = 0.035 for EGFR.31G7 and P = 0.026 for EGFR.25 clone). Because the markers to predict anti-EGFR treatment response in other system tumors such as EGFR mutation and amplification seem to be rare events in breast cancer, membranous staining pattern of EGFR might be the best way to decide the patient eligibility for anti-EGFR therapy.