Reliability of fine needle aspiration biopsy in large thyroid nodules.

OBJECTIVE: Fine needle aspiration biopsy provides one of the most important data that determines the treatment algorithm of thyroid nodules. Nevertheless, the reliability of fine needle aspiration biopsy is controversial in large nodules. The aim of this study was to evaluate the adequacy of fine needle aspiration biopsy in thyroid nodules that are four cm or greater. MATERIAL AND METHODS: We retrospectively examined 219 patients files who underwent thyroidectomy for thyroid nodules that were greater than four centimeter between May 2007 and December 2012. Seventy-four patients with hyperthyroidism, and 18 patients without preoperative fine needle aspiration cytology were excluded from the study. Histopathologic results after thyroidectomy were compared with preoperative cytology results, and sensitivity and specificity rates were calculated. RESULTS: False-negativity, sensitivity and specificity rates of fine needle aspiration biopsy of thyroid nodules were found to be 9.7%, 55.5%, and 85%, respectively. Within any nodule of the 127 patients, 28 (22.0%) had thyroid cancer. However, when only nodules of at least 4 cm were evaluated, thyroid cancer was detected in 22 (17.3%) patients. CONCLUSION: In this study, fine needle aspiration biopsy of large thyroid nodules was found to have a high false-negativity rate. The limitations of fine-needle aspiration biopsy should be taken into consideration in treatment planning of thyroid nodules larger than four centimeters.

Importance of Clinical Decision Making by Experienced Pediatric Surgeons When Children Are Suspected of Having Acute Appendicitis: The Reality in a High-Volume Pediatric Emergency Department.

OBJECTIVE: The aim of the present study was to compare the pediatric appendicitis score (PAS), the Alvarado score (AS), white blood cell count (WBC), absolute neutrophil count (ANC), C-reactive protein (CRP) level, procalcitonin level, and ultrasound (US) data, with the appendectomy decisions of pediatric surgeons diagnosing acute appendicitis (AA) in a real-life setting; this was a top-level, high-volume pediatric emergency department (PED) in a developing country. METHODS: The study was conducted prospectively between January 2012 and June 2013 in the PED of the Tepecik Teaching and Research Hospital in Izmir, Turkey. The study was observational in nature; no attempt was made to influence indications for exploration or the timing thereof. Children aged 4 to 18 years presenting to the PED on suspicion of AA were included. The WBC, ANC, CRP level, and procalcitonin level were measured, and US was performed on all patients on admission. The PAS and AS were calculated. An operative decision was made by each pediatric surgeon who had the results of laboratory and radiological tests. The criterion standard for AA was histopathological assessment. RESULTS: Upon receiver operating curve (ROC) analysis, the areas under the ROCs (AUROCs) of the WBC, ANC, CRP level, procalcitonin level, US positivity, PAS, AS, and decisions of pediatric surgeons supported by laboratory and US data were 0.734, 0.741, 0.671, 0.675, 0.670, 0.831, 0.794, and 0.910, respectively. When US data were employed only in cases with PASs 4 to 7, the sensitivity increased but specificity decreased. The sensitivity and specificity of pediatric surgical decisions were 100% and 82.50%, respectively. The difference between the PAS AUROC and the pediatric surgeon decision-making AUROC was significant (P = 0.0393; 95% confidence interval, 0.0470-0.226). CONCLUSIONS: Good pediatric surgical decision making supported by laboratory and US data for those suspected of AA may be the most effective diagnostic tool in a high-volume PED in a developing country.

Congenital Complete Tracheal Ring in a Neonate: A Case Report.

Abnormalities of the upper airway tract lead to congenital high airway obstruction and may complicate neonatal airway management in the delivery room. Congenital complete tracheal rings are a rare and unusual tracheal anomaly, usually presenting in the neonate or infant as respiratory distress. The clinical presentation can vary from almost asymptomatic patients to near-fatal airway obstruction. It may exist as an isolated entity, or in association with other congenital malformations, in particular, cardiac anomalies along with vascular rings and pulmonary slings. Other associated anomalies have also been reported, for example, chromosomal anomalies, malformation of other parts of the respiratory tract, esophagus and skeletal systems. Here, we report an extreme case of VACTERL/TACRD association presented with congenital complete tracheal ring, encephalocele, bilateral radial agenesis with absent thumbs, equinovalgus deformity on right foot, low-set ears and micrognathia.

Investigating KRAS/BRAF mutation in oropharyngeal squamous cell carcinomas: a preliminary study.

OBJECTIVES: This study aims to investigate the role of KRAS/BRAF gene mutation in the pathogenesis of oropharyngeal squamous cell carcinoma (OSCC). PATIENTS AND METHODS: A total of 26 OSCC patients (23 males, 3 females; mean age 60 years; range 41 to 77 years) diagnosed between January 2003 and November 2013 were included in the study. The methods used in our study were quantitative fluorescence polymerase chain reaction for KRAS/BRAF mutation analysis. RESULTS: Ten of the tumors were located at the tongue base, 12 in the tonsil and four at the floor of mouth. The mean tumor size was 3.8 cm. Six of the tumors were well differentiated, 18 were moderately differentiated and two were poorly differentiated. All cases were analyzed for KRAS and BRAF gene mutations and none of them showed gene mutations. CONCLUSION: We could not find any relation between OSCC and KRAS/BRAF gene mutations in our short case file. The role of mutations should be analyzed in larger series in OSCC to predict new targeted therapy modalities.

Does single receptor positive breast carcinoma differ with regard to age, tumor grade, and HER2 amplification status?

PURPOSE: To investigate whether there is a difference in patient and tumor characteristics in cases with single receptor positive (SRP) (ER-/PR+ and ER+/PR-) breast carcinoma in comparison with the double receptor positive (DRP) (ER+/PR+) and double receptor negative (DRN) (ER-/PR-) tumors. METHODS: A total of 255 breast cancer patients were categorized on the basis of their tumor hormonal receptor phenotype, age, grade, and HER2 amplification status. The study focused on the SRP phenotype (ER+/PR- and ER-/PR+) and compared it with the DRP (ER+/PR+rpar; and DRN (ER-/PR-) tumors. RESULTS: There were 103 (40.3%) DRP tumors, 98 (38.4%) DRN tumors and 54 (21%) SRP tumors, 41 (16.1%) of which were ER+/PR- and 13 (5.1%) were ER-/PR+. Compared to DRP tumors, the SRP group was more likely to be associated with grade 3 tumors and higher frequency of HER2 amplification status. ER-/PR+ tumors were more likely to be associated with younger age at diagnosis compared to ER+/PR- tumors. HER2 amplification, age, and grade were not significantly different between ER-/PR+ and DRN groups. Compared to the DRN group, the ER+/PR- group had lower grade. CONCLUSIONS: Our findings demonstrated that SRP phenotype including ER+/PR- and ER-/PR+ tumors is different from DRP group with regard to age, grade, and HER2 amplification status. Moreover, our data showed that ER-/PR+ tumors are associated with younger age.

Bladder cancer diagnosis from bladder wash by Fourier transform infrared spectroscopy as a novel test for tumor recurrence.

This study proposes Fourier Transform Infrared (FTIR) spectroscopy as a more sensitive, rapid, non-destructive and operator-independent analytical diagnostic method for bladder cancer recurrence from bladder wash than other routinely used urine cytology and cystoscopy methods. A total of 136 patients were recruited. FTIR spectroscopic experiments were carried out as a blind study, the classification results of which were then compared with those of cytology and cystoscopy. Firstly, 71 samples (n = 37; bladder cancer and n = 34; control) were studied with transmittance FTIR spectroscopy. After achieving successful differentiation of the groups, to develop a more rapid diagnostic tool and check the reproducibility of the results, the work was continued with different samples (n = 65 as n = 44; bladder cancer and n = 21; control), using the reflection mode (ATR) of FTIR spectroscopy by a different operator. The results revealed significant alterations in moleculer content in the cancer group. Based on the spectral differences, using transmittance FTIR spectroscopy coupled with chemometrics, the diseased group was successfully differentiated from the control. When only carcinoma group was taken into consideration a sensitivity value of 100% was achieved. Similar results were also obtained by ATR-FTIR spectroscopy. This study shows the power of infrared spectroscopy in the diagnosis of bladder cancer.

Natural Killer-Like T-Cell Lymphoma Localized to the Terminal Ileum: Case Report.

Intestinal intraepithelial lymphocytes are non-organized lymphoid populations that are composed of heterogeneous subsets with diverse ontogeny and phenotypes, and the differential diagnosis is crucial. A 43-year-old male patient underwent an emergency laparotomy due to a perforated mass of the terminal ileum. A right hemicolectomy plus small bowel resection was performed. Histopathological examination showed medium to large cells with vesicular nuclei, including marked nucleoli with large, colorless cytoplasm. No signs of celiac disease were found in the adjacent mucosa. The tumor cells were immunohistochemically CD45+, CD3+, CD4+, CD8+, CD56+, Pan-Cytokeratin-, CD20-, CD79a-, CD5- and CD30-. Endomysial antibody and antigliadin antibody, IgM and IgG tests; and anti-Ebstein Barr virus latent membrane protein all proved negative. Finally, the histopathological diagnosis of tumor mass was natural killer-like T-cell lymphoma. Primary intestinal cytotoxic natural killer-like T-cell lymphoma is a rare entity, which is difficult to distinguish from other T-cell lymphomas. In addition to microscopic evaluation, immunohistochemical analysis and serological tests are essential to reach a definitive diagnosis.

Primary squamous cell carcinoma of the urethral diverticulum mimicking prostate cancer: Case report and review of the literature.

Primary urethral carcinomas are uncommon, with urothelial carcinoma as the most common subtype. Urethral diverticulum is also rarely seen in men. A 44-year-old male presented with voiding symptoms. Abdominoperineal resection, prostatectomy, bladder neck excision, and proximal urethral excision were performed. A pathological examination revealed a well-differentiated squamous cell carcinoma (SCC) located inside an urethral diverticulum. We report this unusual case because primary SCC of the male urethral diverticulum is extremely rare. To our knowledge, our patient is only the second reported case.

Determination of HER2 and p53 Mutations by Sequence Analysis Method and EGFR/Chromosome 7 Gene Status by Fluorescence in Situ Hybridization for the Predilection of Targeted Therapy Modalities in Immunohistochemically Triple Negative Breast Carcinomas in Turkish Population.

Triple negative breast cancer (TNBC), an agressive subtype accounts nearly 15 % of all breast carcinomas. Conventional chemotherapy is the only treatment modality thus new, effective targeted therapy methods have been investigated. Epidermal growth factor receptor (EGFR) inhibitors give hope according to the recent studies results. Also therapeutic agents have been tried against aberrant p53 signal activity as TNBC show high p53 mutation rates. Our aim was to detect the incidence of mutations/amplifications identified in TNBC in our population. Here we used sequence analysis to detect HER2 (exon 18-23), p53 (exon 5-8) mutations; fluorescence in situ hybridization (FISH) method to analyse EGFR/chromosome 7 centromere gene status in 82 immunohistochemically TNBC. Basaloid phenotype was identified in 49 (59.8 %) patients. EGFR amplification was noted in 5 cases (6.1 %). All EGFR amplified cases showed EGFR overexpression by immunohistochemistry (IHC). p53 mutations were identified in 33 (40.2 %) cases. Almost 60 % of the basal like breast cancer cases showed p53 mutation. Only one case showed HER2 mutation (exon 20:g.36830_3). Our results showed that gene amplification is not the unique mechanism in EGFR overexpression. IHC might be used in the decision of anti-EGFR therapy in routine practice. p53 mutation rate was lower than the rates reported in the literature probably due to ethnic differences and low sensitivity of sanger sequences in general mutation screening. We also established the rarity of HER2 mutation in TNBC. In conclusion EGFR and p53 are the major targets in TNBC also for our population.

Clinical, demographic and histopathological prognostic factors for urothelial carcinoma of the bladder.

INTRODUCTION: Our aim is to evaluate the influence of clinical and histopathological parameters, including age, gender, tumor stage, grade, tumor differentiation, necrosis, lymphovascular/perineural invasion (LVI/PNI) and concomitant carcinoma in situ (CIS), on outcomes of patients with urothelial carcinoma of the bladder (UCB). MATERIAL AND METHODS: A total of 84 patients who underwent radical cystectomy (RC) (n = 11) and radical cystoprostatectomy (n = 73) for muscle-invasive bladder cancer at our hospital between 2007-2013, were included in the study. RESULTS: The mean age of patients at diagnosis was 66.1, of whom 75 were males and 9 were females. Of the 84 patients, 38 were ≤65 years and 46 were >65 years. Mean tumor diameter was 3.66 cm. There were 38 cases which showed divergent differentiations. Concomitant CIS was observed in 30 tumors, 41 cases showed tumor necrosis, 44 PNI and 61 LVI. The rate of overall survival (OS) in patients aged ≤65 years was statistically significantly higher than in those aged >65 years. A negative statistical relationship was found between OS with lymph node metastasis (LNM) and tumor differentiation. On the other hand, necrosis did not remain significant on multivariate analysis. No statistically significant relationship was found between smoking, tumor stage, PNI, LVI and concomitant CIS and OS. CONCLUSIONS: In this study, advanced age, LNM, tumor differentiation were found to be independent prognostic risk factors associated with OS after RC. These additional factors, which may explain the different clinical course in patients with similar tumor stage and lymph node status, should be taken into consideration in treatment planning.

Anterior apical cores in the initial prostate biopsy does not increase detection of significant prostate cancer.

PURPOSE: To examine the effect of routine sampling anterior apical cores in the initial prostate biopsy among patients that 14-cores of prostate biopsy (PB) planned. MATERIALS AND METHODS: Five-hundred twenty-eight patients with increased prostate-specific antigen (PSA) levels and/or abnormal digital rectal examination underwent transrectal ultrasound and initial PB between November 2012 and October 2013. We performed routine 12-cores extended PB, plus 2 anterior apex samples that were taken from the junction of urethra and apex of the prostate. Site-specific and unique cancer detection rate, tumor characteristics, the presence of clinically insignificant prostate cancer (PCa) (clinical stage ≤ T1, serum PSA level of < 10 ng/mL, biopsy Gleason score ≤ 6, number of positive biopsy cores ≤ 3 and no core with > 50% involvement) and biopsy-related pain were evaluated. RESULTS: PCa was detected in 147 of 451 patients (32.6%). The lateral base of the prostate was the most affected area with 128 of 451 patients (28.3%), followed by unique cancer detection, with 17 of 40 patients (43.5%). Anterior apex (n = 6) was in third place after the lateral apex (n = 8). The patients diagnosed by anterior apex cores were all clinically insignificant PCa. The cancer diagnosis rate would be 31% if 12-cores biopsy was used, but the rate was found to be 32.6% in 14-cores biopsy (P = .016). Average biopsy pain, right anterior apex biopsy pain, and left anterior apex biopsy pain were found to register at 0.61, 1.06 and 1.08 points in the visual analog scale pain score, respectively. When right and left anterior apex biopsy pain is compared to average biopsy pain, the pain level was found to be statistically significantly higher in the biopsies of right and left anterior apex (P = .040 and P = .042, respectively). CONCLUSION: The gold standard for the diagnosis of PCa is at least 8 cores PB. According to our results, although most PCa diagnosis is carried out with 14-cores PB, it should not be forgotten that these patients might have clinically insignificant PCa.

Problems In Determining Her2 Status In Breast Carcinoma.

OBJECTIVE: Human epidermal growth factor receptor 2 (HER2) oncoprotein is overexpressed in 15-25% of breast carcinomas and associated with poor outcome. Assessment of HER2 status accurately is important to select patients who will benefit from targeted therapy. MATERIALS AND METHODS: In this study immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) were used to determine the HER2 status in 308 breast carcinoma cases of which 129 were consultation. The major problems in determining HER2 status and the reasons of discordant results between methods were discussed. RESULTS: HER2 expression was (-) in 124, (+) in 29, (++) in 92, (+++) in 63 cases. 25 of 76 cases consulted as (++) were evaluated as (++) and 15 of 35 cases consulted as (+++) were evaluated as (+++). HER2 amplification was found in 88 (28.6%) of 308 cases by FISH. 3 of 124 (-), 1 of 29 (+), 22 of 92 (++), 62 of 63 (+++) cases were amplified by FISH. The relation between HER2 expression and amplification was statistically significant (p<0.001). Centromere 17 (CEN 17) region amplification was noted in 11 cases of which 2 were (+++), 9 were (++). 6 of the 11 cases showed focal low level, 1 of them showed diffuse high level amplification. CONCLUSION: The concordance rate between IHC (+++) cases and FISH was 95.4% for consultation cases, 100% for our cases. The final concordance rate for both case groups was 98.4%. The possible reasons of discrepancy were triple negativity, preanalytical and analytical procedures of consultation cases and trucut samples.

Isolated metastasis of uterine leiomyosarcoma to the pancreas: Report of a case and review of the literature.

INTRODUCTION: Metastatic tumors of the pancreas are uncommon and rarely detectable clinically. Metastases to the pancreas are rare. We present a patient with pancreatic metastases from a leiomyosarcoma of the uterus and review the literature about the clinical features of pancreatic metastasis and its surgical management. PRESENTATION OF CASE: A 40-year-old woman, who underwent hysterectomy, left oophorectomy, omentectomy and lymp node dissection for leiomyosarcoma of the uterus. At the follow up, the patient complained of non-specific abdominal discomfort. Preoperative diagnosis were pancreatic pseudocyst, cystadenoma or cystadenocarcinoma. At laparotomy, a cystic mass was found in the tail of the pancreas which was invased to the transverse colon mesenterium and the spleen. Distal pancreatectomy with splenectomy and transverse colon resection was performed. Histologically, the tumor was evaluated as poorly differentiated leiomyosarcoma. DISCUSSION: Metastatic lesions of the pancreas are uncommon and less than 2% of all pancreatic malignancies. However a few cases of leiomyosarcoma with metastases to the pancreas have been reported in the literature. Before deciding that the lesion in the pancreas was metastasis, primary leiomyosarcoma of the pancreas had to be ruled out. Histologically, leiomyosarcoma of the pancreas contains interlacing spindle cells with varying degrees of atypia and pleomorphism. The surgical approach to the pancreatic metastases must be aimed complete excision of the tumor with a wide negative margin of clear tissue and maximum preservation of pancreatic remnant if possible. CONCLUSION: In the absence of widespread metastatic disease, aggressive surgical approach with negative margins must be aimed.

Huge congenital haemangioma of the tongue.

Haemangiomas, the most common type of benign vascular tumours, are rare in the oral cavity. Some of these lesions are congenital and show symptoms in late childhood or early adult life. A 32-years-old woman presented with a huge lesion on her tongue which caused dysphagia and dysphasia. She had first noticed the lesion when she was 6. Her obstructive symptoms started when she was 28 and, despite various medical treatments, the size of the lesion gradually increased. Magnetic resonance imaging revealed a 7 x 5 x 3 cm mass on the right side of the tongue. Because of severe functional and cosmetic problems, the lesion was excised with partial haemiglossectomy. Histopathological examination was consistent with intramuscular haemangioma. Haemangiomas are benign tumours with a benign course and are rarely seen on the tongue. They have clinical importance when localised in the oral cavity. Different treatment modalities exist, but in cases of large tumours, surgery may be the mainstay treatment.

Importance of P53, Ki-67 expression in the differential diagnosis of benign/malignant phyllodes tumors of the breast.

BACKGROUND: Conventionally growth pattern, stromal overgrowth, stromal cellularity and stromal mitotic activity are the main parameters in the grading of phyllodes tumors (PTs). Recent studies revealed that both p53 and Ki-67 expressions are correlated with grade of PTs of the breast. Expression of hormone receptors and overexpression/amplification of HER2 has been studied in PTs to discover the roles of these markers as new treatment modalities. MATERIALS AND METHOD: We studied 26 PT cases. Seventeen benign and nine malignant PTs were re-evaluated as regards stromal cellularity mitotic activity, p53/Ki-67 expression rates and the relation between these parameters. Estrogen receptor and progesterone receptor (ER, PR) positivity were determined by counting nuclear staining in five high-power fields. Also, the presence of any HER2 staining and staining patterns were documanted. RESULTS: Stromal cellularity, mitotic rate, p53 and Ki-67 expression rates were all correlated with benign and malignant histologic subgroups (P = 0.000-0.001). Ki-67 and p53 expressions were statistically significantly correlated with histologic subgroups, stromal cellularity and mitotic rate (P < 0.005). ER and PR expressions in the epithelial component were not statistically significant between the two groups. HER2 showed different staining patterns in the epithelial component, and there was no staining in the stromal component. CONCLUSION: Ki-67 and p53 expression rates were statistically significantly correlated with grade of mammary PTs; therefore, they can be used in the determination of tumor grade, especially for the differential diagnosis of benign and malignant tumors. Malignant and benign tumors did not differ significantly in terms of hormone receptor and HER2 expression. HER2 expression showed different patterns in the epithelial component of the PTs.

HER2 status in gastric cancer: a comparison of two novel in situ hybridization methods (IQ FISH and dual color SISH) and two immunohistochemistry methods (A0485 and HercepTest™).

In contrast to breast HER2 testing, the optimal ISH method and antibody for gastric HER2 testing are unclear. The aim of this study was to find out gastric HER2 positivity rates in our institutional data, and to compare the two novel ISH methods with A0485 antibody and HercepTest™. IHC and ISH were carried out on gastrectomy specimens of 88 patients up to the standardly advised procedure protocols, and interpretations were also carried out up to widely accepted international protocols., HER2 expression was (-) in 65, (+) in 5, (++) in 6, and (+++) in 12 cases by A0485 IHC. IHC (+) 4 cases and (++) 3 cases were (-) by HercepTest™. One IHC (-) amplified case was (++) by HercepTest™. All A0485 and HercepTest™ (+++) 12 cases were amplified by ISH. HER2 amplification was detected in 18 (20.4%) and in 15 (17.2%) cases by SISH and FISH, respectively. Of the 18 cases, 4 showed focal heterogeneous low level amplification by SISH. Focal amplification was noted in only 2 cases by FISH. The HER2 status of our gastric cancer file is 17.2% by FISH, 20.4% by SISH. The concordance between HercepTest™/A0485 IHC and ISH is perfect in (+++) cases. Equivocal results (++) with any IHC method should be clarified by one of the molecular methods (SISH and FISH). Probably up to the higher level of heterogeneity of gastric carcinomas, there is a 4.5% dilemma of cases that are negative or weakly positive by conventional IHC methods. Therefore, regarding HER2 status in gastric carcinoma, the reliability of IHC methods should be checked.

An unusual case of trisomy 18 associated with paucity of bile ducts.

A case of neonatal cholestasis associated with Trisomy 18 (Edward's syndrome) is presented. A 3-day-old boy was referred to our clinic due to respiratory distress, elevated serum direct bilirubin levels, a systolic heart murmur, growth restriction and micrognathia. Liver biopsy and chromosomal analysis revealed paucity of intrahepatic bile ducts and Trisomy 18. Extrahepatic biliary atresia was reported in only a few patients with Trisomy 18. To our knowledge, we described for the first time a patient with Trisomy 18 and neonatal cholestasis associated with paucity of interlobular bile ducts.

Paratesticular sarcomas: our case series.

OBJECTIVE: Paratesticular tumors, which comprise a heterogeneous group of entities, are often described in case reports in the literature. In this study, we present histomorphological, immunohistochemical and clinical features of six cases with paratesticular sarcoma. MATERIAL AND METHODS: Six cases with paratesticular sarcoma diagnosed in our hospital between 1997 and 2012 were included in this study. Information regarding treatment modalities, tumor recurrence, metastasis, and survival were obtained from archival patient records. Hematoxylin-eosin sections of the cases were examined, and immunohistochemical analyses were performed for markers including smooth muscle actin, desmin, Ki67, CD34, S100 and myogenin. Percentage of staining in five high-power fields were counted to document Ki67 and p53 nuclear positivity rates. RESULTS: Of the 6 paratesticular sarcoma cases, 3 were rhabdomyosarcomas, 2 leiomyosarcomas and 1 liposarcoma. The case with sclerotic-type liposarcoma showed two recurrences during the 15-year follow-up period. Two cases with the diagnosis of leiomyosarcoma presented with lung metastases at the time of diagnosis, and 1 patient died of the disease at 7(th) month. Of the 3 cases with rhabdomyosarcomas, 2 patients were lost to postoperative follow up. The other patient presented with liver and prevertebral metastasis at the 3(rd) month and died of the disease in the 14(th) month. The Ki67 proliferation index was significantly higher for one case with rhabdomyosarcoma, and 2 cases with leiomyosarcoma. Differences in p53 expression were not statistically significant between the cases. CONCLUSION: Paratesticular tumors belong to a heterogeneous group of tumors that can follow different clinical courses. This study showed that the most important features in determining prognosis are histopathological subtype and tumor grade.