High Mitotic Index Research Articles

The Effect of the COVID-19 Lockdown on Melanoma Diagnosis in Italy

The coronavirus disease 2019 (COVID-19) pandemic has led to lockdowns for much of the world. In Italy, all health procedures not directly related to COVID-19 were reduced or suspended, thus limiting patient access to hospitals. Any delay in cancer treatment presents the additional risk of tumors progressing from being curable to incurable. Specifically, melanoma survival rate strictly depends on tumor thickness, which, in turn, is a function of time. To estimate the impact on melanoma progression caused by the reduction in dermatologic services during the COVID-19 lockdown, a retrospective observational cohort study was conducted. This study was designed to compare the clinical and histologic characteristics of the primary melanomas removed in the first 2 months after the end of the lockdown (May-July 2020) in 12 Italian centers characterized by different COVID-19 case frequencies. The control group was represented by the melanomas removed during the same period in the previous 3 years. Overall, 1,124 melanomas were considered: 237 as part of the study group and 887 from the control group (average, 295), with a 20% reduction. Breslow thickness, as well as high-risk histotypes and melanomas with vertical growth, increased for all melanomas. Ulcerated and high mitotic index melanomas increased, particularly in northern Italy. In Italy, the lockdown led to a significant worsening of melanoma severity, causing a staging jump, with a consequent worsening of outcomes.

Histiocytic sarcoma arising from a lymph node: a diagnostic conundrum

Histiocytic Sarcoma (HS) is extremely rare, with only a few hundred cases reported in the literature. The majority of patients present with symptoms due to unifocal or multifocal extra-nodal disease. Less than 20% of these cases show solitary involvement of a lymph node. We report a case of a solitary HS in a 53-year-old woman presenting with a 2.7-cm right groin mass arising from an inguinal lymph node. The initial cytologic examination of the tissue showed a high-grade spindle-shaped morphology with high-grade mitotic activity. A high-grade sarcoma was initially considered considering the absence of normal lymphoid aggregate and the presence of high-grade cytologic features in the cells. To evaluate the tumor in its entirety, the mass was surgically excised. A histological examination of the tumor showed focal rimming of the lymphoid tissue at the periphery and a centrally located stellate necrotic focus. The tumor cells had an epithelioid to spindle cell morphology along with large uniform nuclei and prominent nucleoli. A high mitotic index was present. Immunohistochemistry (IHC) stains showed strong positivity for CD68, CD163, and Vimentin, and were weakly positive for SMA and CD45. Based on the histologic and clinical examination, a diagnosis of HS was made. Multiple malignancies can mimic HS histopathology and the rarity of this tumor makes the diagnosis more challenging. No fine-needle aspiration (FNA) criteria for its diagnosis have been recognized. Herein, we report a rare case of an isolated HS involving a lymph node which resembled high-grade sarcoma on the FNA biopsy to raise awareness among our surgical pathologist colleagues.

Adrenocortical oncocytic tumors: a clinicopathological analysis of 44 cases

Objective: To study the clinicopathological features of adrenocortical oncocytic tumors (ACOT) and to compare the diagnostic values of Lin-Weiss-Bisceglia (LWB) score and Helsinki score. Methods: Forty-four cases of ACOT diagnosed at Beijing Friendship Hospital, China from March 2008 to July 2019 were histologically analyzed to evaluate their malignant potential (benign versus malignant) according to two scoring criteria. Immunohistochemical studies (EnVision method) were also used. Results: There were 23 males and 21 females with an average age of 46 years. Histologically, the tumor cells were arranged in trabecular, chrysanthemum-shaped, glandular and microcapsule structures, while clear cells were rare or absent. Most of the tumor cells were moderately atypical, and intranuclear inclusion bodies were conspicuous. Immunohistochemical staining showed that tumor cells were positive for Melan A, inhibin, Syn and calretinin. The average proliferation index was 3% in benign ACOT, about 5% in ACOT of malignant potential, and>20% in malignant ACOT. According to the LWB score, 61.4% (27/44) of the tumors were on the left side and had multiple lesions. The percentage of benign ACOT was 59.1% (26/44), malignant potential 6.8% (3/44), malignant 34.1% (15/44), respectively. Among the 15 malignant ACOT, the mitotic figures>5/50 HPF were found in 13 cases, necrosis in 11 cases and capsule invasion in 10 cases. According to the Helsinki score, 65.9% (29/44) of the tumors were benign, and 34.1% (15/44) were malignant. There was no significant difference between the two scoring standards (P>0.05). During the follow-up of 9 to 144 months, 31 patients survived without disease and 13 patients relapsed or had metastasis. Conclusions: ACOT more likely be benign than malignant. The left side is more common. Malignant tumors are prone to recurrence and metastasis. The morphological parameters (high mitotic index, necrosis, and capsular invasion) in the LWB scoring standards combined with immunohistochemical parameters (Ki-67) in the Helsinki score are helpful for the diagnosis of malignant ACOT and are important predictors of poor prognosis.

Pathological and Cytological Studies on Hepatocellular Carcinoma in Cattle Slaughtered at Bishoftu Elfora Abattoir, Central Ethiopia.

Hepatocellular carcinoma (HCC) is one of the most common neoplasms that has been described in many domestic animal species. Hence, the disease has significant economic importance; thus, this study aimed to describe the cytopathological characteristics of hepatocellular carcinomas in cattle slaughtered at Bishoftu Elfora Abattoir, Central Ethiopia. A cross-sectional study design with a purposive sampling technique was performed from October 2017 to May 2018 using macroscopic, histopathologic, and cytological methods. For that matter, a total of sixty cattle were assessed for the presence of a hepatic tumor; however, only 1/60 (0.6%) case was found to be affected by hepatocellular carcinomas. On gross examination, hepatocellular carcinomas exhibited soft, white, multifocal nodules (10–40 mm in diameter) on different lobes of the liver. On the cut surface, the tumor revealed a sharply circumscribed border and was divided into lobules by thin connective tissue. The central zone of the tumors exhibited depression with a whitish fibrous area. Moreover, on histopathology, the tumors divulged unencapsulated carcinomatous lesions consisting of a thick, compact, somewhat ambiguous trabecular pattern of arrangement that was unglued by thin collagenous stroma. Cytological studies suggest that the tumor cells showed anisocytosis, anisokaryosis, prominent nucleoli, multinuclearity, palisading arrangements of neoplastic cells, increased N : C ratios, light eosinophilic cytoplasm, high mitotic index, and cytoplasmic and intranuclear vacuoles. In conclusion, cytopathological findings support a diagnosis of HCC in the liver; thus, further studies with a large sample size and use of immunohistochemistry are important for further characterization of hepatocellular carcinomas in cattle.

Preoperative C-Reactive Protein/Albumin Ratio is a Prognostic Indicator for Survival in Surgically Treated Gastrointestinal Stromal Tumors: A Retrospective Cohort Study

BackgroundSystemic inflammation and malnutrition may promote tumor progression. C-reactive protein/albumin ratio (CAR) is linked to the poor long-term survival of several malignant tumors.PurposeTo explore the predictive value of CAR in gastrointestinal stromal tumors (GISTs).MethodsA retrospective study was conducted on 325 patients with primary GIST surgically treated with curative intent from 2009 to 2018. The cut-off point of CAR was set using X-tile software. Kaplan–Meier method and multivariate Cox regression model were used to study the prognostic value of CAR. The time-dependent receiver operating characteristic curve (tROC) was drawn, and the prognostic accuracy of CAR, Glasgow prognostic score (GPS), and National Institute of Health (NIH) risk classification was compared by the area under the curve (AUC).ResultsThe best cut-off point of CAR was 0.55. Increased CAR was associated with the location of the lower digestive tract, larger tumor size, higher mitotic index, higher NIH risk classification, lower ALB, higher CRP, and higher GPS (all p<0.05). Multivariable analysis revealed that CAR (hazard ratio [HR] 2.598, 95% confidence interval [CI] 1.385–4.874; p=0.003) was an independent predictor of overall survival. Additionally, the AUC of CAR was lower than that of NIH risk classification at 2 years (0.601 vs. 0.775, p=0.002) and 5 years (0.629 vs 0.735, p=0.069). However, the AUC of NIH risk classification significantly increased (2-year OS 0.801, p=0.251; 5-year OS 0.777, p=0.011) when combined with CAR.ConclusionCAR is a new independent predictor of poor survival in patients with GIST. CAR combined with NIH risk classification can effectively improve the performance of prognosis prediction.

Stanford parameters stratify the risk of recurrence in gynecologic smooth muscle tumors of uncertain malignant potential.

Smooth muscle tumor of uncertain malignant potential (STUMP) is an ill-defined category of neoplasms, which represent a diagnostic challenge. We aimed to assess whether the Stanford parameters, that is, high mitotic index (≥10/10HPF), significant atypia (moderate-to-severe), and coagulative tumor cell necrosis (CTCN), even when focal or ambiguous, may be used to stratify the risk of recurrence in gynecological smooth muscle tumor of uncertain malignant potential (STUMP). Electronic databases were searched from their inception to October 2019. All studies assessing the Stanford parameters in gynecological STUMP series were included. STUMPs were subdivided according to the presence of the three Stanford parameters: high mitotic index, significant atypia, and CTCN. A Kaplan-Meier survival analysis was performed for recurrence-free survival; hazard ratio (HR) was calculated in each category. Fourteen studies with 219 STUMPs were included. In 15.5% of cases, none of the three Stanford parameters were present, with a recurrence risk of 5.9%; 2.7% of cases showed high mitotic index alone, with a recurrence risk of 0% (HR = not calculable); 43.8% of cases showed significant atypia alone, with a recurrence risk of 18.7% (HR = 3.3; p=0.012); 26.5% of cases showed CTCN alone, with a recurrence risk of 17.2% (HR = 3.1; p=0.029); and 11.4% of cases showed at least two Stanford parameters, with a recurrence risk of 32% (HR = 7.5; p=0.003). Stanford parameters may stratify the risk of recurrence of STUMP. Significant atypia and CTCN, but not high mitotic index, may be stand-alone risk factors for recurrence in STUMP. The presence of at least two Stanford parameters, even if equivocal (e.g., uncertain or focal CTCN, focal significant atypia, mitotic index around 10/10HPF), might still be enough to support a diagnosis of leiomyosarcoma. Further studies are necessary in this field.

Effective Treatment for Primary Locally Aggressive Intermediate and Malignant Soft Tissue Tumors of the Breast

ObjectiveThis study aimed to examine the rare locally aggressive intermediate tumors and malignant primary breast mesenchymal tumors in patients receiving surgical treatment.Summary of background dataLocally aggressive intermediate tumors were subdivided into nonmetastasizing and rarely metastasizing, and the malignant group was subdivided as a single subgroup called able to metastasize. A retrospective examination of surgical notes and clinical charts was carried out reviewing gender, age, symptoms, duration of symptoms, tumor size, clinical presentation, radiation history, kind of surgery undergone, adjuvant radiotherapy, adjuvant chemotherapy, local recurrences, systemic metastases, and mortality.ResultsMitotic index and Ki-67 were statistically different between locally aggressive and malignant groups (P &amp;lt; 0.001). One local recurrence occurred in only 1 patient diagnosed as dermatofibrosarcoma protuberans among all patients. None of the entities showed distant metastasis or mortality. In this result, clear margin of surgery in locally aggressive intermediate group and combination of surgery with radiation therapy in the malignant group was the most important determinant for the prospect of the patients with mean follow-up of 28 months.ConclusionThe main treatment for localized mesenchymal breast tumors is surgery. When the disease is locally advanced or malignant with high mitotic index, radiotherapy with surgery is predominantly used. Targeted therapies are promising with the limited place of chemotherapy.

Correlation of Endoscopic Ultrasonography Features with the Mitotic Index in 2- to 5-cm Gastric Gastrointestinal Stromal Tumors

Background: Predicting the malignancy potential of gastrointestinal stromal tumor (GIST) before resection could improve patient management strategies as gastric GISTs with a low malignancy potential can be safely treated endoscopically, but surgical resection is required for those tumors with a high malignancy potential. This study aimed to evaluate endoscopic ultrasound (EUS) features of 2- to 5-cm gastric GISTs that might be used to predict their mitotic index using surgical specimens as the gold standard. Patients and Methods: Forty-nine patients (30 females and 19 males; mean age 55.1 ± 12.7 years) who underwent EUS examinations, followed by surgical resections of 2- to 5-cm gastric GISTs, were retrospectively reviewed. Results: The mean tumor size was 3.44 ± 0.97 (range 2.1–5.0) cm. A univariate analysis revealed no significant differences in age, sex, and tumor location in the low mitotic index and high mitotic index groups (all p > 0.05). In terms of EUS features, there were no significant differences in the mitotic indexes with respect to the shape, surface lobulation, border regularity, echogenicity, homogeneity, growth patterns, presence of mucosal ulceration, hyperechogenic foci, anechoic spaces, and hypoechoic halos (all p > 0.05). However, the tumor size was larger in the high mitotic index group than that in the low mitotic index group (3.97 ± 1.05 vs. 3.27 ± 0.9 cm, p = 0.03). Conclusion: Conventional EUS features are not reliable for predicting the mitotic index of 2- to 5-cm gastric GISTs. Further modalities for predicting the mitotic index are needed to prevent unnecessary surgical resections in patients with a low risk of malignancy.

Evaluation of Asa River Water in Ilorin, Kwara State, Nigeria for Available Pollutants and their Effects on Mitosis and Chromosomes Morphology in Allium cepa Cells

T: This study evaluated water samples from Asa River in Kwara state, Nigeria, for cytogenotoxicity at 25.0 %, 50.0 %, 100.0 % following the Allium cepa assay. Onions were grown in the water samples for microscopic and macroscopic screenings. Heavy metals and volatile organic pollutants in the water were elucidated using AAS and GCMS techniques. The Water samples except the sample ‘C’ induced higher mitotic index (MI) than the negative control. Root growth was significantly promoted at 25.0 %, and significantly reduced at 50.0 % and 100.0 % of the sample ‘C’ and 100.0 % of the sample ‘A’. Water sample ‘A’ (100.0%) induced highest percentage chromosomal aberrations (CA) while the water samples ‘B’ and ‘C’ induced higher percentage CA than the negative control. Cadmium was detected at a concentration higher than its permissible limit in drinking water. Poly aromatic hydrocarbons, Aromatic amines, Acridine dye, Phenolic and Polychlorinated compounds were detected in the water sample. The observed proliferative, inhibitory, cytotoxic and genotoxic effects of the water samples on A. cepa cells suggest that Asa river was polluted, having potential to adversely affect humans, animals and plants utilizing it along its course.&#x0D; Keywords: Allium cepa; Asa River; cytogenotoxicity; Nigeria; pollutants

Neoadjuvant therapy for gastrointestinal stromal tumors: A propensity score‐weighted analysis

Retrospective and single arm prospective studies have reported clinical benefit with neoadjuvant imatinib for GISTs. In the absence of randomized phase III data, the impact of neoadjuvant systemic therapy (NAT) on survival compared to upfront resection (UR) remains unknown. We identified N=16,308 patients within the National Cancer Database (2004-2016) who underwent resection of localized GIST of the stomach, esophagus, small bowel, and colorectum, with or without ≥3 mon of NAT. Inverse probability of treatment weighting adjusted for covariable imbalance among treatment groups. We estimated the effect of NAT on overall survival with a weighted time-dependent Cox proportional hazards model, and on 90-day postoperative mortality and R0 resection with weighted logistic regressions. 865 (5.3%) patients received NAT, compared to 15,443 (94.7%) who underwent UR. Median NAT duration was 6.3 months. 53.7% of NAT patients were male vs. 48.6% of UR patients, 67.3% vs. 65.1% had primary gastric GIST, and 72.8% vs 49.7% were high-risk. NAT patients had larger tumors and higher mitotic index. >3 months of NAT was associated with a significant survival benefit (weighted HR 0.85 (0.80-0.91)). 90-day postoperative mortality rate was 4/865 (0.5%) among NAT patients vs. 346/15,443 (2.2%). NAT was associated with lower odds of 90-day postoperative mortality. R0 resection rate was not significantly different between groups. In conclusion, despite higher risk features among NAT patients, this analysis suggests that NAT for localized GIST is associated with a modest survival benefit and lower risk of 90-day postoperative mortality, with no difference in likelihood of achieving an R0 resection. This article is protected by copyright. All rights reserved.

Intussusception caused by intestinal neoplasia in mature rainbow trout (Oncorhynchus mykiss, Walbaum 1792).

The aquaculture industry is growing and includes the farming and breeding of more than 580 aquatic species worldwide. The rainbow trout (Oncorhynchus mykiss, Walbaum 1792) is the most commonly bred trout species in Hungary. As broodfish form the basis of most fish farms, investigation into tumours occurring in trout, although under-researched, has proven to be a valuable and necessary field of study. During our investigation, we examined a broodstock of 3- to 6-year-old rainbow trouts (800) affected with idiopathic intestinal tumours (3%) which had consequentially led to ileus (40%). While performing necropsy, initial pathological observations showed intussusceptions. Tumours were discovered upon opening the body cavity, as well as metastasis forming in the livers and in the vessels of the gills. Histopathological and immunohistochemical tests allowed us to identify the neoplasms. The primary adenocarcinoma was found to have been developed within the intestines of the fish. The tumour tissue broke through the basal membrane and infiltrated the propria, protruding asymmetrically into the lumen of the mid-intestines, causing it to narrow significantly. This subsequently led to passage disorders, invagination of the intestinal segment and finally the emaciation of the fish. Histopathological and immunohistochemical inspection of the tumour cells displayed a high mitotic index, confirming malignancy.

Integrating Anatomical, Molecular and Clinical Risk Factors in Gastrointestinal Stromal Tumor of the Stomach

BackgroundAdjuvant imatinib for 3 years is recommended to patients with high-risk gastrointestinal stromal tumor (GIST). Risk stratification is inaccurate, and risk assessments are further complicated by the increased use of neoadjuvant treatment. Anatomical criteria for prognostication have not been investigated.MethodsClinical, molecular, and anatomical variables were retrospectively studied in a population-based cohort of 295 patients with gastric GIST resected between 2000 and 2018. Gastric subsite was divided into the upper, middle, and lower thirds. Growth pattern was classified as luminal, exophytic, or transmural based on imaging and surgical reports.ResultsOf 113 tumors in the upper third of the stomach, 103 (91.2%) were KIT mutated, 7 (6.2%) were PDGFRA mutated, and 104 (92.0%) harbored genotypes sensitive to imatinib. Transmural tumors were strongly associated with a high mitotic index. Five-year recurrence-free survival (RFS) was 71% for patients with transmural tumors versus 96% with luminal or exophytic tumors (hazard ratio [HR] 8.45, 95% confidence interval [CI] 3.69–19.36; p < 0.001), and, in high-risk patients, 5-year RFS was 46% for patients with transmural tumors versus 83% with luminal or exophytic tumors (HR 4.47, 95% CI 1.71–11.66; p = 0.001). Among 134 patients with tumors > 5 cm, there were 29 recurrences. Only five patients with exophytic or luminal tumors had recurrent disease, of whom four had tumor rupture. Five-year RFS for patients with exophytic/luminal tumors >5 cm without rupture was 98%.ConclusionsIn the upper third, over 90% of tumors were sensitive to imatinib. Patients with exophytic or luminal tumors without rupture, irrespective of size, had an excellent prognosis and may not benefit from adjuvant therapy.

Spindle Cell Melanoma Harboring a Nodule of Epitheloid Cell Melanoma Component: A Study of a Diagnostically Challenging Case

Abstract Background: Melanoma is a very heterogeneous human neoplasm. In addition to four major (conventional) histologic subtypes a number of uncommon variants do exist. Objective: An unusual case of a spindle cell melanoma (SCM) containing a demarcated nodule of conventional epitheliod cell melanoma component is described. Material and Methods: A 71-year-old man manifested with a protuberated ulcerated skin tumor arising on the right forearm. The resected biopsy was analyzed immunohistochemically with a variety of anti-human antibodies. Results: The tumor consisted of a highly cellular mass of spindle-shaped cells without any significant intratu-moral fibrosis. In addition, a nodule of epithelioid cell tumor component was present within the lesion. The spindle cell component showed a disperse reactivity for S100 protein and was negative for other melanocytic markers. It exhibited a very high mitotic activity and proliferation Ki-67 index. No melanin pigment was detected. In contrast, the epithelioid cell component was strongly positive for S100 protein, Melan-A/MART-1, HMB-45, and PNL-2. The mitotic and proliferation indices were much less pronounced and melanin deposits were visible. A diagnosis of a non-desmoplastic SCM harboring a nodule of epithelioid cell melanoma component was established. Conclusion: SCM often posses a diagnostic dilemma because its histomorphology is atypical and its immunohistochemical profile may differ from other subtypes of melanomas. The present paper points out this uncommon histopathological entity that may sometimes be encountered in dermatopathological practice and that requires more complex diagnostic approach.

Loss of BRCA1 expression and morphological features associated with BRCA1 promoter methylation status in triple-negative breast cancer.

Aberrant DNA methylation in the BRCA1 promoter region causes epigenetic silencing of BRCA1 gene expression, which is critical for breast cancer development. However, how BRCA1 promoter methylation status alters histological features remains poorly understood. Here, we investigated the possibility to predict BRCA1 promoter methylation status based on the morphological and immunohistochemical features of triple-negative breast cancers (TNBCs). The morphological features of 53 TNBCs were evaluated with hematoxylin-eosin staining, with immunohistochemical staining of BRCA1, androgen receptor, p53, cytokeratin 5/6, and epidermal growth factor receptor. BRCA1 promoter methylation status was used to distinguish BRCA1 promoter-methylated tumors (BPMTs) from BRCA1 promoter-unmethylated tumors (BPUTs) dependent on pathological characteristics. BPMTs comprised approximately 26% of the TNBCs. Immunohistochemical analysis found that BRCA1 protein expression was significantly lower in BPMT compared with BPUT (p = 0.016). Morphologically, BPMTs were associated with high mitotic index (p = 0.017), pushing margin (p = 0.017), a circumscribed growth pattern (p = 0.014), and a syncytial growth pattern (p = 0.034) compared with BPUTs. We then assessed the potential of predicting BRCA1 promoter methylation status by using published score systems based on these morphological characteristics. A receiver operating characteristic analysis showed an area under the curve of 0.80. This study found that BRCA1 promoter methylation status could be derived from morphological features and lower BRCA1 expression of TNBCs, which may help identify suitable cases for target treatment with PARP inhibitors.

Clinicopathological characteristics and outcomes in men with mesothelioma of the tunica vaginalis testis: analysis of published case-series data

PurposeMesothelioma of the tunica vaginalis testis (MTVT) is a rare tumor, and currently, there are no published treatment recommendations.MethodsWe performed a systematic literature review and synthesized clinical presentation, clinicopathological factors associated with metastatic disease, treatment options, and outcomes in men with MTVT.ResultsWe included 170 publications providing data on 275 patients. Metastatic disease occurred in 84/275 (31%) men with malignant MTVT: Most common sites included retroperitoneal lymph nodes (LNs) (40/84, 48%), lungs (30/84, 36%), and inguinal LNs (23/84, 27%).Invasion of the spermatic cord or scrotum was the only risk factor for local recurrence [odds ratio (OR) 3.21, 95% confidence interval (CI) 1.36–7.57]. Metastatic disease was associated with age ≥ 42 years (OR 3.02, 95% CI 1.33–6.86), tumor size ≥ 49 mm (OR 6.17, 95% CI 1.84–20.74), presence of necrosis (OR 8.31, 95% CI 1.58–43.62), high mitotic index (OR 13.36, 95% CI 1.53–116.51) or angiolymphatic invasion (OR 3.75, 95% CI 1.02–13.80), and local recurrence (OR 4.35, 95% CI 2.00–9.44). Complete remission in the metastatic setting was observed in five patients, most of whom were treated with multimodal therapy. Median survival in patients with metastatic disease was 18 months (IQR 7–43).ConclusionMalignant MTVT is a rare but aggressive disease. Since local recurrence is a risk factor for metastatic progression, we recommend aggressive local treatment. Survival and response to any treatment in the metastatic setting are limited.

PARP1 as a Marker of an Aggressive Clinical Phenotype in Cutaneous Melanoma-A Clinical and an In Vitro Study.

(1) Background: Poly(ADP-ribose) polymerase 1) (PARP1) is a pleiotropic enzyme involved in several cellular processes, e.g., DNA damage repair, regulation of mitosis, and immune response. Little is known about the role of PARP1 in melanoma development and progression. We aimed to investigate the prognostic significance of PARP1 expression in cutaneous melanoma through evaluation of mRNA and protein levels of PARP1 in normal melanocytes and melanoma cell lines, as well as in patients’ tissue material from surgical resections. (2) Methods: An in vitro model was based on two types of normal human melanocytes (HEMn-DP and HEMn-LP) and four melanoma cell lines (A375, WM1341D, Hs294T, and WM9). PARP1 mRNA gene expression was estimated using real-time polymerase chain reaction (RT-PCR), whereas the protein level of PARP1 was evaluated by fluorescence confocal microscopy and then confirmed by Western Blotting analysis. The expression of PARP1 was also assessed by immunohistochemistry in formalin-fixed paraffin-embedded tissues of 128 primary cutaneous melanoma patients and correlated with follow-up and clinicopathologic features. (3) Results: The in vitro study showed that melanoma cells exhibited significantly higher PARP1 expression at mRNA and protein levels than normal melanocytes. High PARP1 expression was also associated with the invasiveness of tumor cells. Elevated nuclear PARP1 expression in patients without nodal metastases strongly correlated with significantly shorter disease-free survival (p = 0.0015) and revealed a trend with shorter cancer-specific overall survival (p = 0.05). High PARP1 immunoreactivity in the lymph node-negative group of patients was significantly associated with higher Breslow tumor thickness, presence of ulceration, and a higher mitotic index (p = 0.0016, p = 0.023, and p < 0.001, respectively). In patients with nodal metastases, high PARP1 expression significantly correlated with the presence of microsatellitosis (p = 0.034), but we did not confirm the prognostic significance of PARP1 expression in these patients. In the entire analyzed group of patients (with and without nodal metastases at the time of diagnosis), PARP1 expression was associated with a high mitotic index (p = 0.001) and the presence of ulceration (p = 0.036). Moreover, in patients with elevated PARP1 expression, melanoma was more frequently located in the skin of the head and neck region (p = 0.015). In multivariate analysis, high PARP1 expression was an independent unfavorable prognosticator in lymph node-negative cutaneous melanoma patients. (4) Conclusions: In vitro molecular biology approaches demonstrated enhanced PARP1 expression in cutaneous melanoma. These results were confirmed by the immunohistochemical study with clinical parameter analysis, which showed that a high level of PARP1 correlated with unfavorable clinical outcome. These observations raise the potential role of PARP1 inhibitor-based therapy in cutaneous melanoma.

Neuroendocrine Carcinomas of the Canine Mammary Gland: Histopathological and Immunohistochemical Characteristics.

Invasive mammary carcinomas with neuroendocrine differentiation are rare in women and were reported only once in female dogs. For the present study, ten cases of solid mammary carcinoma positive for chromogramin A in immunohistochemistry were selected. Histopathological characteristics of these tumors were described and immunohistochemical evaluation was performed with chromogranin A, synaptophysin, CD56, NSE, PGP 9.5, pancitokeratin, Ki67, estrogen receptor (ER), and progesterone receptor (PR). The average animal age was 13.2 years old and the average tumor size was 4.8 cm. In total, 70% of the neoplasms were classified as grade III and 30% as grade II by the Nottingham histological grade system. High mitotic index was observed with a mean of 27.5 mitoses in 10 high magnification fields. Only one case showed typical carcinoid tumor characteristics. In addition, vascular invasion was shown in 3 tumors. All carcinomas were positive for chromogran A, while only two cases were reactive to synaptophysin. For PGP 9.2, NSE and CD56, we observed positivity of 100, 90, and 70%, respectively, in the samples, being that no tumor was positive for all the neuroendocrine markers. All neoplasms showed ER and PR in at least 10% of neoplastic cells, while Ki67 varied from 29 to 95%, with mean mitotic index of 67%. Four of the ten animals died within 1 year of the tumor diagnosis. Neuroendocrine neoplasms occur in the canine mammary gland and are propably underdiagnosed. This is due to their non-specific morphological characteristics and the low use of neuroendocrine immunohistochemistric markers the diagnostic routine. More studies are necessary to determine the prognosis of this new histological type.

A Rare Case of Pediatric Gastrointestinal Stromal Tumor

Aim: To discuss the management of rare pediatric tumor, Gastrointestinal Stromal Tumor (GIST)Methods: We report a case of 1 year old male child with transverse colonic GIST. Child presented with upper abdominal lump of one month duration with no other constitutional symptoms. On examination solitary intra-abdominal lump of size 10cm and 8cm, non tender, firm, freely mobile in the umbilical region was found. Contrast enhanced CT abdomen revealed 8.3cm and 6.2cm heterogenous enhancing mass at root of the mesentery. On exploration an exophytic pedunculated mass was arising from mid transverse colon, tumor excised along with 2 cm colonic margin. Counselling done as chemotherapy was not available in children. Child has been under regular followup for the last 18 months. Results: Histopathological examination revealed GIST with high mitotic index, immuno-histochemistry (IHE) markers CD117,CD34 and DOG 1 were positive.Conclusion: Pediatric GIST is very rare and there are no well established therapeutic guidelines for the management of pediatric GIST, so needs close long term followup after surgery and further guidelines need to be established.

Significance of cyclin D1 overexpression for the diagnosis of mantle cell lymphoma: a clinicopathologic comparison of cyclin D1-positive MCL and cyclin D1-negative MCL-like B-cell lymphoma

AbstractMantle cell lymphoma (MCL) is a distinct clinicopathologic entity of non-Hodgkin’s lymphoma, characterized by a monotonous proliferation of small to medium-sized lymphocytes with co-expression of CD5 and CD20, an aggressive and incurable clinical course, and frequent t(11;14)(q13;q32) translocation. We examined 151 cases of lymphoma with MCL morphology from a viewpoint of cyclin D1 overexpression, which is now easily detectable by immunohistochemistry. 128 cases (85%) showed positive nuclear staining for cyclin D1, while the remaining 23 (15%) were negative. Except for cyclin D1 immunohistochemistry, current diagnostic methods, including morphological and phenotypical examinations, could not make this distinction. Although both the cyclin D1-positive and -negative groups were characterized by male predominance, advanced stages of the disease, frequent extranodal involvement, and low CD23 reactivity, the cyclin D1-positive group showed a higher age distribution (P = .04), larger cell size (P = .02), higher mitotic index (P = .01), more frequent gastrointestinal involvement (P = .05), higher international prognostic index score (P = .05), and lower p27KIP1 expression (P < .0001). Of particular interest is that cyclin D1-positive MCL showed significantly worse survival than cyclin D1-negative lymphoma (5-year survival: 30% versus 86%, P = .0002), which was confirmed by multivariate analysis to be independent of other risk factors. These data suggest that cyclin D1-positive and -negative groups may represent different entities and that the former closely fits the characteristics of classical, typical MCL. We therefore propose that cyclin D1-positivity should be included as one of the standard criteria for MCL, and that innovative therapies for this incurable disease should be explored on the basis of the new criteria. (Blood. 2000;95:2253-2261

Co-delivery of EGFR and BRD4 siRNA by cell-penetrating peptides-modified redox-responsive complex in triple negative breast cancer cells

AimsTriple negative breast cancer (TNBC) has drawn more and more attention due to its high mitotic indices, high metastatic rate and poor prognosis. Gene therapy, especially RNA interference (RNAi), has become a promising targeted therapy. However, improvement of transfection efficiency and discovery of target genes are major problems for the delivery of small interfering RNAs (siRNA). Materials and methodsIn the present study, we developed GALA- and CREKA-modified PEG-SS-PEI to deliver siRNAs targeting on EGFR and BRD4 for TNBC therapy. The PEG-SS-PEI/siRNA complexes were prepared by electrostatic interaction and characterized by dynamic light scattering (DLS) and transmission electron microscope (TEM). The release characteristic, stability, cellular uptake and intracellular localization of the complexes were also studied. The effect of the complexes on cell viability was measured in MDA-MB-231 and HUVEC cells. The in vitro anti-tumor activities of the complexes were analyzed by Transwell invasion assay and wound healing assay. The gene silencing effect was evaluated by quantitative real time-polymerase chain reaction (qRT-PCR) and western blot. Key findingsThe results revealed that the GALA- and CREKA-modified PEG-SS-PEI/siRNA complexes showed excellent transfection efficiency with redox-sensitive release profile and good biological compatibility. The complexes protected siRNA from the degradation of RNA enzymes. The complexes significantly inhibited the proliferation, invasion and migration of MDA-MB-231 cells via the synergistic inhibition of EGFR/PI3K/Akt and BRD4/c-Myc pathways. SignificanceTaken together, co-delivery of siEGFR and siBRD4 by GALA-PEG-SS-PEI and CREKA-PEG-SS-PEI may provide a more effective strategy for the treatment of TNBC.