Immunohistochemical evaluation of lymphovascular invasion in carcinoma breast with CD34 and D2-40 and its correlation with other prognostic markers.

BACKGROUND: Carcinoma breast is ever-evolving and becoming increasingly prevalent in India. Numerous prognostic factors based on morphology and immunohistochemistry (IHC) have been established which need to be interconnected to give patients best possible treatment. AIMS: This study aims to confirm and analyze lymphovascular invasion (LVI) detected by hematoxylin and eosin (H and E) using IHC with CD34 and D2-40 and its correlation with other biologic and morphologic prognostic markers. SETTINGS AND DESIGN: This was a prospective study. MATERIALS AND METHODS: Fifty mastectomy specimens diagnosed as infiltrating ductal carcinoma breast on histopathology selected for the study. Evaluation of formalin-fixed paraffin-embedded sections was done using H and E and IHC for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 HER2/neu receptors, CD34, and D2-40 endothelial markers. Correlation of LVI done with prognostic markers of Carcinoma Breast, namely, age of the patient, tumor size, Nottingham grade, lymph node ratio (LNR), Nottingham prognostic index (NPI), ER/PR status, and HER2/neu status. CD34 and D2-40 utilized to distinguish blood vessel, lymph vessel, and retraction artifacts and to calculate lymphatic microvessel density (LMVD) and blood microvessel density (BMVD). STATISTICAL ANALYSIS USED: SPSS Software Package. RESULTS: LVI was associated with younger age (P = 0.001), greater tumor size (P = 0.007), higher Nottingham grade (P = 0.001), higher LNR (P = 0.001), higher NPI (P = 0.001), Negative ER Status (P = 0.001), Negative PR Status (P = 0.002), Positive HER2/neu status (P = 0.021), Higher Intratumoral BMVD (P = 0.016), Peritumoral BMVD (P = 0.001), and Intratumoral LMVD (P = 0.009). Blood vessels more commonly invaded than lymph vessels. Retraction artifacts can be mistaken for LVI without IHC. CONCLUSIONS: D2-40 is a promising marker for lymphatic endothelium. LVI is a poor prognostic marker hence should be evaluated imperatively in all cases of carcinoma breast.

Evaluation of Histologically and Histochemically Proven Cases of Vitiligo and its Correlation with CD4+ and CD8+ Lymphocyte Counts using Flow Cytometry.

INTRODUCTION: Vitiligo is an acquired cutaneous disorder characterized by progressive and selective destruction of melanocytes from the epidermis. Autoimmunity is strongly implicated in its pathogenesis. The destruction of melanocytes has a correlation with the peripheral blood lymphocyte imbalance mainly including Cytotoxic T cells (CD8+cells) and Helper T cells (CD4+cells). The progression of vitiligo is associated with higher CD8+ counts and lower CD4+ counts thus, altering CD4+: CD8+ ratio. AIM: To evaluate the clinically suspected cases of vitiligo histopathologically and histochemically and to establish the co-relation of autoimmunity with the flow cytometric analysis of CD8+ and CD4+ lymphocyte counts. MATERIALS AND METHODS: In this study, 40 patients with proven vitiligo were taken. The destruction of melanocytes was confirmed by Haematoxylin & Eosin (H & E) and by histochemical stains using Dihydroxyphenylacetic Acid (DOPA) reaction. Blood sample from these vitiligo patients and 10 control subjects was taken. Flow Cytometry was used for the determination of CD8+ and CD4+ counts in the vitiligo patients and control subjects. Then CD4+: CD8+ ratio was calculated and comparison between vitiligo patients and control subjects was done. T-test was used for the statistical analysis. RESULTS: There was statistically significant decrease in CD4+: CD8+ ratio. CD4+: CD8+ ratio was decreased in 57.5% cases of vitiligo with increase in CD8+ counts and decreased CD4+ counts. CONCLUSION: It was concluded from this study that cellular immunity might have a role to play in the pathogenesis of vitiligo causing the destruction of melanocytes.

Primary Extra-Gonadal Retroperitoneal Seminoma, Clinically Mimicking Sarcoma.

Extra-gonadal Germ cell tumours (EGCTs) are germ cell tumours that are found in locations other than the gonads. These tumours are seen in young males in the midline, mostly in the mediastinum followed by retroperitoneum, pineal gland, and sacrococcygeal area. While most of these are non-seminomatous, few are seminomatous. We present here a case report of a 25-year-old, mentally retarded male patient, with bilaterally normal testes, who presented with a huge mass in abdomen. The ultrasound and CECT (Contrast Enhanced Computerized Tomography) scan done showed two large retroperitoneal masses, which on FNAC (Fine Needle Aspiration Cytology) suggested germ cell tumour. Surgical resection was done and the subsequent histopathological examination provided the diagnosis of seminoma, which was further confirmed by PAS (Periodic Acid Schiff) staining and Immunohistochemistry for PLAP (Placental Alkaline Phosphatase). Retroperitoneum is a common site for sarcomas, which also present as huge, bulky masses. Thus, EGCTs should be considered in the differential diagnosis of a soft-tissue mass in abdomen, especially arising in the retroperitoneum.

Primary subcutaneous inguinal hydatid cyst: diagnosis by fine needle aspiration cytology.

Hydatid disease or human cystic echinococcosis, recognized by ancient scholars such as Hippocrates, Galen and Rhazes, is one of the oldest diseases known to man. Though hydatid cyst may develop in almost any part of the body, a solitary primary subcutaneous localization is an extremely rare entity. We herein report a case of primary subcutaneous inguinal hydatid cyst which was diagnosed by fine needle aspiration cytology. Radiological examination done subsequently corroborated with the cytodiagnosis of Hydatid cyst and did not show involvement of any other organ or site. Histopathological examination of surgically removed cyst confirmed the diagnosis of Hydatid cyst. Hydatid disease should be considered in the differential diagnosis of all unusual swellings in soft tissues, especially in regions where the disease is endemic.

To evaluate the role of bone marrow aspiration and bone marrow biopsy in pancytopenia.

BACKGROUND: Pancytopenia is not a disease entity but a triad of findings that may result from various disease processes, primarily or secondarily involving the bone marrow. Bone marrow aspiration and biopsy evaluation along with good clinical correlation is of utmost importance to evaluate the causes of pancytopenia and planning further investigations. AIMS: The present study was a prospective clinicohaematological study undertaken to analyse the various causes of pancytopenia by evaluating bone marrow aspiration and biopsy and correlating with clinical findings, complete blood counts and peripheral blood picture. MATERIALS AND METHODS: Fifty patients of pancytopenia were included in the study in which relevant history and physical examination findings were recorded. Bone marrow aspiration and biopsy were performed simultaneously in all cases. Perl's stain was done in all cases and special stains like MPO, PAS and reticulin were also done wherever necessary. RESULTS AND CONCLUSION: The maximum cases of pancytopenia were in the age group of 10 to 30 y with male preponderance. Aplastic anaemia was found to be the most common aetiology of pancytopenia followed by normoblastic erythroid hyperplasia, megaloblastic anaemia, acute leukemias, myelofibrosis, lymphoid neoplasia and iron deficiency anaemia. It was concluded from the study that although the advantages of bone marrow aspiration and biopsy differ, both are complimentary to each other and should be performed simultaneously for a complete bone marrow work up and evaluation. It is only through the correlation of clinical, hematological and bone marrow examination findings that proper evaluation and management of patients of pancytopenia can be made.

Primary endometrial squamous cell carcinoma with extensive squamous metaplasia and dysplasia.

Primary squamous cell carcinoma of endometrium is a rare entity. Only 64 cases have been documented in the literature. We report a case of 60-year-old postmenopausal woman who presented with abdominal distention and blood-stained vaginal discharge for 6-7 months. Clinically, chronic pyometra was considered. Total abdominal hysterectomy was performed and histopathologically, it was diagnosed as a case of primary squamous cell carcinoma of endometrium with extensive squamous metaplasia and dysplasia.

Seroprevalence of hepatitis C antibodies in healthy blood donors--a prospective study.

In the last decade, hepatitis C has emerged from obscurity as a disease (Non A Non B Hepatitis)familiar to only a few experts, to being recognized as a major public health problem. The present study was done to find out the seroprevalence of hepatitis C virus in 5000 blood donors in Patiala and to compare its seroprevalence in voluntary and replacement blood donors. The testing for HCV antibodies was done by ELISA technique using third generation HCV microlisa kit. Seroprevalence of anti-HCV amongst 5000 healthy blood donors was 0.88%. It was 0.58% in voluntary blood donors and 0.95% in replacement blood donors. Anti-HCV positivity was significantly higher amongst males (0.97%) than females (0.59%). Maximum anti-HCV positivity was seen in 41-50 year age group in voluntary donors and 31-40 year age group in replacement group. This study made us to conclude that above results were due to multiple reasons like transfusion of blood and blood products from unscreened donors, medical injections, tattooing, intravenous drug abuse, traditional medicinal practices, sexual promiscuity and lack ofawareness in rural population about the disease and mode of spread.