Role of WT1, B-cell lymphoma 2, Ki-67 (Mib1), and Her2/Neu as diagnostic and prognostic immunomarkers in ovarian serous and endometroid carcinoma.

BACKGROUND: Ovarian cancer is the fifth common cause of death due to cancer in women. It constitutes 3% of all cancers in females and 15%-20% of genital malignancy. Most of the ovarian cancers are serous type followed by the endometrioid type. Sometimes, glands of these two carcinomas are indistinguishable histologically. It also becomes difficult to differentiate these two types when they are poorly differentiated. AIMS: The aim of this study was to find differences in immunomarker expressions between serous and endometrioid carcinomas and the association of their staining patterns with other clinicopathological prognostic factors. MATERIALS AND METHODS: Immunohistochemical staining for WT1, B-cell lymphoma 2 (Bcl2), Ki67 (MIB1), and Her2/Neu were done in paraffin-embedded tissues of histologically diagnosed 38 cases of ovarian serous and endometrioid carcinomas and staining patterns were correlated with other clinicopathological prognostic factors. RESULTS: Of these 38 cases (21 serous and 17 endometrioid), 24 cases were in Stage I/II and 14 cases Stage III/IV. On the other hand, 16 cases were in low grade, 12 cases intermediate grade, and 10 cases high grade. Twenty of 21 serous carcinomas were strongly positive for WT1, whereas most of the endometrioid carcinomas were negative. Ki67 (MIB1) labeling index and Her2/Neu were higher in both higher grade and stages. On the contrary, the intensity of Bcl2 staining was lower in higher grades and stages lesions. CONCLUSIONS: The use of WT1 may be useful in resolving diagnostic dilemma between serous and endometrioid carcinoma, especially in difficult cases. Ki67, Bcl2, and Her2/Neu may be used as prognostic markers.

Near-set Based Mucin Segmentation in Histopathology Images for Detecting Mucinous Carcinoma.

This paper introducesnear-set based segmentation method for extraction and quantification of mucin regions for detecting mucinouscarcinoma (MC which is a sub type of Invasive ductal carcinoma (IDC)). From histology point of view, the presence of mucin is one of the indicators for detection of this carcinoma. In order to detect MC, the proposed method majorly includes pre-processing by colour correction, colour transformation followed by near-set based segmentation and post-processing for delineating only mucin regions from the histological images at 40×. The segmentation step works in two phases such as Learn and Run.In pre-processing step, white balance method is used for colour correction of microscopic images (RGB format). These images are transformed into HSI (Hue, Saturation, and Intensity) colour space and H-plane is extracted in order to get better visual separation of the different histological regions (background, mucin and tissue regions). Thereafter, histogram in H-plane is optimally partitioned to find set representation for each of the regions. In Learn phase, features of typical mucin pixel and unlabeled pixels are learnt in terms of coverage of observed sets in the sample space surrounding the pixel under consideration. On the other hand, in Run phase the unlabeled pixels are clustered as mucin and non-mucin based on its indiscernibilty with ideal mucin, i.e. their feature values differ within a tolerance limit. This experiment is performed for grade-I and grade-II of MC and hence percentage of average segmentation accuracy is achieved within confidence interval of [97.36 97.70] for extracting mucin areas. In addition, computation of percentage of mucin present in a histological image is provided for understanding the alteration of such diagnostic indicator in MC detection.

Automated characterization and counting of Ki-67 protein for breast cancer prognosis: A quantitative immunohistochemistry approach.

Ki-67 protein expression plays an important role in predicting the proliferative status of tumour cells and deciding the future course of therapy in breast cancer. Immunohistochemical (IHC) determination of Ki-67 score or labelling index, by estimating the fraction of Ki67 positively stained tumour cells, is the most widely practiced method to assess tumour proliferation (Dowsett et al. 2011). Accurate manual counting of these cells (specifically nuclei) due to complex and dense distribution of cells, therefore, becomes critical and presents a major challenge to pathologists. In this paper, we suggest a hybrid clustering algorithm to quantify the proliferative index of breast cancer cells based on automated counting of Ki-67 nuclei. The proposed methodology initially pre-processes the IHC images of Ki-67 stained slides of breast cancer. The RGB images are converted to grey, L*a*b*, HSI, YCbCr, YIQ and XYZ colour space. All the stained cells are then characterized by two stage segmentation process. Fuzzy C-means quantifies all the stained cells as one cluster. The blue channel of the first stage output is given as input to k-means algorithm, which provides separate cluster for Ki-67 positive and negative cells. The count of positive and negative nuclei is used to calculate the F-measure for each colour space. A comparative study of our work with the expert opinion is studied to evaluate the error rate. The positive and negative nuclei detection results for all colour spaces are compared with the ground truth for validation and F-measure is calculated. The F-measure for L*a*b* colour space (0.8847) provides the best statistical result as compared to grey, HSI, YCbCr, YIQ and XYZ colour space. Further, a study is carried out to count nuclei manually and automatically from the proposed algorithm with an average error rate of 6.84% which is significant. The study provides an automated count of positive and negative nuclei using L*a*b*colour space and hybrid segmentation technique. Computerized evaluation of proliferation index can aid pathologist in assessing breast cancer severity. The proposed methodology, further, has the potential advantage of saving time and assisting in decision making over the present manual procedure and could evolve as an assistive pathological decision support system.

Quantitative microscopic evaluation of mucin areas and its percentage in mucinous carcinoma of the breast using tissue histological images.

Mucinous carcinoma (MC) of the breast is very rare (∼1-7% of all breast cancers), invasive ductal carcinoma. Presence of pools of extracellular mucin is one of the most important histological features for MC. This paper aims at developing a quantitative computer-aided methodology for automated identification of mucin areas and its percentage using tissue histological images. The proposed method includes pre-processing (i.e., colour space transformation and colour normalization), mucin regions segmentation, post-processing, and performance evaluation. The proposed algorithm achieved 97.74% segmentation accuracy in comparison to ground truths. In addition, the percentage of mucin present in the tissue regions is calculated by the mucin index (MI) for grading MC (pure, moderately, minimally mucinous).

Evaluation of vaginal cytology in female sex-workers: A study in a tertiary hospital of eastern India.

BACKGROUND: Papanicolaou (Pap) test is an important and easy diagnostic tool to detect any abnormalities on vaginal cytology. Pap test is routinely done in women of reproductive age group in many countries. AIM: The aim of this study was to detect spectrum of abnormalities in female sex workers (FSWs) on vaginal cytology. MATERIALS AND METHODS: A total of 60 cases were included over a period of 1 year (July, 2011-June, 2012). The age range of the patients was 14-61 years. Pap stained slides were evaluated by two consultant cytopathologists and reported as normal smear, inflammatory smear, specific infection, low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL), atypical squamous cell of undetermined significance (ASCUS), squamous cell carcinoma (SCC) and atypical glandular cell of undetermined significance (AGUS). RESULTS: Most of the smears were abnormal accounting for 86.7% of total cases (52/60). Incidence of HSIL was very high in FSWs. Out of 60 cases, 8 normal smears (13.3%), 12 inflammatory smears (20%), 3 cases of infections (5%), 9 cases of LSIL (15%), 23 cases of HSIL (38.3%), 2 cases (3.3%) each of ASCUS and SCC and 1 case (1.3%) AGUS were encountered. CONCLUSIONS: Close follow-up and histologic examinations are necessary to avoid unnecessary spread of the neoplastic disease and untimely death of the patients. Awareness about diseases in FSWs and diagnostic utility of Pap test are also a must.

Primary pleomorphic rhabdomyosarcoma of breast: Report of a rare neoplasm.

Primary sarcoma of the breast is very rare and constitutes less than 1% of all breast cancers. Herein, we report a case of pleomorphic rhabdomyosarcoma (PRMS) of the right breast in a 49-year-old female patient presented with a mass (7 cm × 6.5 cm). Mammography and ultrasonography suspected a malignant lesion and a diagnosis of poorly differentiated carcinoma was made on fine needle aspiration cytology. Modified radical mastectomy was carried out. Histopathological examination revealed a high grade stromal sarcoma with rhabdoid morphology and multinucleated tumor giant cells. The tumor cells were strongly positive for desmin, vimentin and Myo D1 focally. The tumor cells were immunonegative for cytokeratin, epithelial membrane antigen (EMA), CD34, CD45, SMA, S100, CD68 and HMB45. A final diagnosis of PRMS was rendered. Surgical margins were free and no metastasis was seen in axillary lymph nodes. Neither post-operative radiotherapy nor adjuvant chemotherapy was given and the patient has remained disease free 12 months post-operatively.

The Bethesda system for reporting thyroid fine needle aspirates: A cytologic study with histologic follow-up.

BACKGROUND: The Bethesda system for reporting thyroid cytopathology represents a major step towards standardization, reproducibility, improved clinical significance, and greater predictive value of thyroid fine needle aspirates (FNAs). AIMS: To elucidate the utility of the Bethesda system in reporting thyroid FNAs. MATERIALS AND METHODS: We retrospectively reviewed thyroid FNAs between April 2009 and March 2012, classified them using the Bethesda system, found out the distribution of cases in each Bethesda category, and calculated the malignancy risk for each category by follow-up histopathology. RESULTS: Of the 1020 FNAs, 1.2% were non-diagnostic, 87.5% were benign, 1% were atypical follicular lesion of undetermined significance (AFLUS), 4.2% were suspicious for follicular neoplasm (SFN), 1.4% were suspicious for malignancy (SM), and 4.7% malignant. Of 69 cases originally interpreted as non-diagnostic, 12 remained non-diagnostic after re-aspiration. In 323 cases, data of follow-up histopathologic examination (HPE) were available. Rates of malignancy reported on follow-up HPE were non-diagnostic 0%, benign 4.5%, AFLUS 20%, SFN 30.6%, SM 75%, and malignant 97.8%. CONCLUSIONS: Reviewing the thyroid FNAs with the Bethesda system allowed a more specific cytological diagnosis. In this study, the distribution of cases in the Bethesda categories differed from some studies, with the number of benign cases being higher and the number of non-diagnostic and AFLUS cases being lower. The malignancy risk for each category correlated well with other studies. The Bethesda system thus allows standardization in reporting, improves perceptions of diagnostic terminology between cytopathologists and clinicians, and leads to more consistent management approaches.

Endoscopic Dacryocystorhinostomy: A Study at IPGMER, Kolkata.

Endoscopic sinus surgery has developed a new era in the treatment of chronic dacryocystitis. A prospective study was conducted by taking 67 patients having epiphora. Endo DCR was performed under local anaesthesia. Post operative care was given by means of alkaline nasal douching, lacrimal sac massage and endoscopic examination on 1st, 3rd, 6th week. Success rate was found to be 95.5 % which is comparable to external DCR and with added advantages.

Auricular perichondritis in a tertiary rural hospital

@#<p><strong>OBJECTIVE:</strong> Auricular perichondritis refers to inflammation involving the perichondrium of the external ear. It is a very serious disease which may lead to permanent deformity of the pinna. We describe the predisposing factors, pathogenic organisms, interventions and residual deformities in a prospective consecutive sample of patients.<br /><br /><strong>METHODS:<br />Design:</strong> Prospective Cohort Study<br /><strong>Setting:</strong> Tertiary Rural Government Teaching Hospital<br /><strong>Participants:</strong> All patients presenting with auricular perichondritis for a period of one year between March 2011 and February 2012 were consecutively enrolled and a clinical history and demographic details were obtained. Routine hematologic, blood biochemical examinations and culture / sensitivity of discharge from the pinna were conducted, and empiric intravenous ciprofloxacin was administered and continued if confirmed by culture and sensitivity. Those sensitive to co-amoxiclav, ceftazidime or amikacin were shifted to those medications. Medications were shifted to oral forms when available and indicated by resolution of acute inflammation, wound healing and no growth on cultures. Parenteral medications were maintained until the same parameters were achieved. Surgical incision and drainage was also performed when indicated, followed by a standardized wound care regimen. Follow up was for six months ending with assessment of pinna deformity.<br /><br /><strong>RESULTS:</strong> Of the total study population of 50, 76% were male and 24% were female; 15-75 years of age (range 60 years) displaying male predominance and clustering in the fourth decade of life. The most common predisposing factors were trauma from motor vehicle accidents (30%) followed by high ear piercing (22%). <em>Pseudomonas aeruginosa</em> (48%) followed by <em>Staphylococcus aureus</em> (20%) were the most common organisms isolated. All were managed with intravenous antibiotics but 76% also required surgical intervention. Sixty-eight percent developed residual deformities of the pinna with 50% being total and 18% being partial. <br /><br /><strong>CONCLUSION</strong>: Auricular perichondritis is a frightening disease which requires early management. As <em>Pseudomonas aeruginosa</em> is the most common organism, antipseudomonal antibiotics should be started as early as possible. Despite medical and surgical intervention, residual deformities may ensue.</p>

A study on outcome of underlay, overlay and combined techniques of myringoplasty.

Myringoplasty is a procedure which deals on repair of the tympanic membrane. This procedure can be done via postaural, endaural or endomeatal route. Various grafts such as temporalis fascia, vein graft, perichondrium are used. The technique can be categorized as underlay, overlay, interlay or its combination depending on the placement of the graft material. This study was done to compare underlay, overlay and combined technique in terms of the closure of the membrane defect, postoperative complications and over all success rates. Apart from few complications, this study revealed over all success rate was best with combined technique but the difference was not significant statistically when the methods are comparable among them.

Clinico-epidemiological profile of chronic suppurative otitis media patients attending a tertiary care hospital.

The cross-sectional study was conducted to assess the clinico-epidemiological profile, perceptions and clinical profile of the chronic suppurative otitis media (CSOM) patients in a tertiary care hospital. A pretested questionnaire was used containing open questions and the patients were assessed clinically. Most patients (31.2%) were from 0-10 years age group and were males (58.8%). Majority of them (96%) lived in "kuccha" houses/slums, 76.8% practised unhygienic ear pricking, 36.8% poured oil in their ears, 70.8% bathed in ponds/rivers, 52.8% had ear discharge for more than 1 year. Among the respondents, 17.2% knew that CSOM was contagious, 24% thought CSOM ran in family, 20% knew CSOM is preventable. There was low threat perception and long time to seek care. Patients mostly presented with earache, deafness and discharge, most had deafness and safe variety of CSOM. More than half had comorbidities. Most of the previous study findings corroborated with the present study. Here was a substantial delay between the onset and treatment seeking due to lack of awareness and low threat perception. Pain and complications were the triggers for care-seeking. Education about the disease, strengthening the frontline workers and good referral system are suggested.

An arrow penetrating at base of the skull successfully removed.

A 30-year-old male presented with accidental injury with an arrow which referred to us from a peripheral village hospital. It was found that the arrow was penetrating through the nasal bones. An xray skull lateral view showed the tip of the arrow penetrating into the posterior wall of the sphenoid sinus. As the patient had no clinical evidence of neurological or vascular injury, he was immediately operated upon and the arrow was removed. Patient was discharged in good condition and a 3-month follow-up was normal.

A study of surgical management of chronic suppurative otitis media with cholesteatoma and its outcome.

OBJECTIVE: Aim of this study is evaluation of course of improvement of surgically treated cases of chronic suppurative otitis media (CSOM) with cholesteatoma; it includes hearing status, condition of mastoid cavity, study of different, natural and surgical condition and recurrence of disease within the study period. DESIGN: It is a prospective study. SETTINGS: This study was conducted in a premiere government hospital in Kolkata between May 2007 to April 2008. PATIENTS: Total 40 patients between age group of 6-70 years were included in the present study which includes 19 males and 21 females. INTERVENTION: Surgical interventions were done in all the cases. Different types of mastoidectomy with or without tympanoplasty was done according to extent of disease process. OUTCOME: Audiometrically documentable hearing improvement occurred in 35% cases (p = 14), in rest of the ears hearing status remained unaltered. At the end of 6 months follow up 92.5% (p = 14) in rest (p = 37) operated ears become completely dry. Five percent cases (p = 2) presented with facial paralysis; among them one patient improved completely and another patient improved from grade V to grade III facial paralysis. No patient developed any post operative intracranial complications and recurrence of cholesteatema not found in 6 months follow up. Meatal stenosis developed in 5% cases (p = 2) at the end of 6 months. CONCLUSION: Surgery is mainstay of treatment in CSOM with cholesteatoma. Eradication of disease, prevention of complication, maintenance and restoration of hearing, and giving the patient a non-discharging ear are main aim of treatment.

A study on role of nasal endoscopy for diagnosis and management of epistaxis.

Epistaxis is one of the commonest ENT emergencies. It affects people of all ages, more commonly males. Bleeding may be due to local nasal pathology, systemic diseases, or sometimes no specific cause is found, which is called idiopathic epistaxis. This descriptive study was designed to evaluate role of nasal endoscopy for diagnosis and management of epistaxis in a tertiary care hospital. Sixty-two selected patients with active epistaxis or with history of epistaxis within last 24 hours were evaluated and underwent nasal endoscopy. Patients were managed following standard protocols. This study demonstrated a bimodal distribution with incidence peaks in below 20 years and above 50 years age groups. Males were affected nearly twice commonly as females. Anterior nasal bleeding was noted in majority. Most common cause was found to be hypertension closely followedby trauma. Nasal endoscopy helped to localise bleeding points in majority of the cases. Cases were managed accurately with the help of endoscopes and any local disease was eradicated thoroughly. During follow-up, endoscopy helped in early detection of recurrences and thereby prevented complications. Epistaxis can be controlled very efficiently by electro or chemical cauterisation with the help of endoscopes, the source can be localised more efficiently. In cases of failure to localise or access of bleeding points, anterior and/or posterior nasal packing can control majority of nosebleeds. In majority of cases non-surgical interventions were sufficient.

Clinicopathological study of otogenic brain abscess.

Chronic suppurative otitis media (CSOM) is one of the commonest disease entities encountered in otolaryngology practice. Due to poor economic conditions poor hygiene lack of education and death of knowledge about the disease and its complications, we frequently encounter patients of CSOM with extracranial and intracranial complications. Among the intracranial complications otogenic brain abscess is one of the dreadest. With the improvement of healthcare and accessibility of the poor people to healthcare system and development of CT scan, MRI as diagnostic tools incidence of otogenic brain abscess is becoming less due to early diagnosis and interventions. The objectives of this study are to assess the incidence of brain abscesses to explore age, sex, variation to study clinical presentations and common pathogens responsible.