Correlation of light and electron microscopic morphometric parameters of glomerular capillaries with serum creatinine and proteinuria.

Waste products in the bloodstream are filtered by the glomerular capillaries in the kidneys and excreted into the urine. When making a differential diagnosis of kidney diseases, structural assessment of glomeruli using histological, ultrastructural, and immunological studies is crucial. This study assessed the microscopic and ultrastructural morphometric parameters of glomerular capillaries and examined their correlation with serum creatinine and proteinuria. A total of 60 kidney biopsy cases received by the transmission electron microscope (TEM) laboratory for diagnosis were included in the study. Toluidine blue stained 300 nm thick sections of TEM tissue blocks were scanned for glomerular morphometry by a whole slide imaging system, and the estimation of Bowman's capsule (BC) area, glomerular capillary lumen diameter (GCLD), glomerular capillary density (GCD), glomerular capillary surface area density (GCSA), and percentage of glomerular capillary lumen space (%GCLS) was performed with QuPath software. TEM images of 70 nm thick sections were used for the evaluation of endothelial fenestration diameter (EFD), glomerular basement membrane (GBM) thickness, and podocyte foot process (PFP) effacement. Proteinuria and serum creatinine showed positive correlations with GBM thickness and PFP effacement. Negative correlations of serum creatinine were observed with EFD, %GCLS, and GCSA. Hence, glomerular filtration is greatly affected by the total area of the glomerular capillary surface and structural changes of GBM. Reduction of glomerulus filtration due to foot process effacement and thickening of GBM results in damage to the filtration barrier leading to the leakage of plasma protein into urine.

Bisphosphonate Osteonecrosis in a Case of Langerhans Cell Histiocytosis: Report of a Case and Review of Literature.

Langerhans Cell histiocytosis is a rare multisystem disease originating from dendritic cell lines primarily characterized by osteolytic lesion of the skeleton. Chemotherapy along with bisphosphonates have been used to control the progression of the disease and prevent the osteolysis associated with the disease. Bisphosphonate osteonecrosis of the jaws is a common finding following treatment with bisphosphonates for osteoporosis, metastatic tumors to the bone or the osteolytic lesions of multiple myeloma. The article highlights a rare case of bisphosphonate osteonecrosis of the mandible in a case of long-standing Langerhans cell histiocytosis.

Myeloid sarcoma: an uncommon presentation of myeloid neoplasms; a case series of 4 rare cases reported in a tertiary care institute.

Myeloid sarcoma (MS) is a rare extramedullary neoplasm of myeloid cells, which can arise before, concurrently with, or following hematolymphoid malignancies. We report 04 such cases of MS, diagnosed in this institute over a period of 6 years, during various phases of their respective myeloid neoplasms/leukemias. These cases include MS occurring as a relapse of AML (Case 1), MS occurring as an initial presentation of CML (Case 2), MS occurring during ongoing chemotherapy in APML (Case 3), and MS presenting as a progression of MDS to AML (Case 4). In the absence of relevant clinical history and unemployment of appropriate immunohistochemical (IHC) studies, these cases have a high risk of being frequently misdiagnosed either as Non-Hodgkin's Lymphoma (NHL) or small round cell tumors or undifferentiated carcinomas, which may further delay their management, making an already bad prognosis worse. This case series has been designed to throw light on the varied presentation of MS and the lineage differentiation of its neoplastic cells through the application of relevant IHC markers along with their clinical correlation.

Comparative performance analysis of 2D and 3D gamma metrics for patient specific QA in VMAT using Octavius 4D with 2D-Array 1500.

INTRODUCTION: Gamma pass percentage (GPP) is the predominant metric used for Patient Specific Quality Assurance (PSQA) in radiation therapy. The dimensionality of the measurement geometry in PSQA has evolved from 2D planar to 3D planar, and presently to state-of-the-art 3D volumetric geometry. We aim to critically examine the performance of the three-dimensional gammas vis-à-vis the older gamma metrics of lower dimensionality to determine their mutual fungibility in PSQA, using clinically approved Volumetric Arc Therapy (VMAT) plans. METHODS AND MATERIALS: Gamma pass percentages derived from PSQA for VMAT plans using Octavius 4D phantom with 2D-Array 1500 and its proprietary software were recorded. 2D planar, 3D planar, and 3D volumetric gamma pass percentages were retrospectively extracted for multiple treatment plans at three sites, using three acceptance limits, and for two modes of normalization. The differences in mean pass percentages, and the pairwise correlation between geometries were calculated within limits of statistical significance. RESULTS: A significant increase in mean pass rates was observed from 2D planar to 3D planar geometries. The difference was less pronounced from 3D planar to 3D volumetric. 2D planar v/s 3D planar showed a significant degree of correlation among themselves, which was not seen against most of the 3D volumetric pass rates. CONCLUSION: The mean gamma pass rates show conclusive evidence of the benefits of shifting from 2D planar to higher dimensions measurement geometries, but the benefits of using 3D volumetric compared to 3D planar is not always unequivocal. The correlations show mixed results regarding the interdependence of pass percentages at different geometries.

Clinical audit of dose-escalated radical radiotherapy for advanced cervical carcinoma using a pragmatic protocol (3 fractions of 8 Gy HDR brachytherapy).

INTRODUCTION: Recent image-guided brachytherapy data suggests, dose-escalation to a cumulative EQD2 (equivalent dose delivered at 2 Gy/#) of ≥87 Gy is associated with significantly better disease control. We present a clinical audit of a pragmatic radical radiotherapy protocol for advanced cervical cancer, using fewer fractions of brachytherapy than in the presently most popular protocol. MATERIAL & METHODS: Between July 2015 and December 2018, 96 consecutive advanced cervical carcinoma patients were treated by pelvic external beam radiotherapy (EBRT) (50 Gy/25fractions/5 weeks) ± weekly intravenous chemotherapy followed by image guided high dose rate (HDR) brachytherapy, using intracavitary/interstitial/hybrid techniques (intended point A dose: 8 Gy/fractions) × 3 fractions (cumulative target EQD2 ≥ 86 Gy). Insertion was done individually for each fraction of treatment. RESULTS: All patients completed their intended radiation protocol. 93.8% patients achieved complete response, while 6.2% patients achieved only partial response; no patients had stable/progressive disease. Out of the patients with partial response, 4.2% (4 out of 5 cases) cases of central/nodal residual disease underwent salvage surgery. At a median follow up of 21 months, 8.3% (8) patients had local failure, 1.1% (1) had nodal failure and 3.1% (3) had distant failures. Median Failure Free Survival was 29 months (26.5-31.5 months). On follow up, 6.3% and 3.2% patients had grade 2 or worse rectal and bladder morbidities respectively. CONCLUSION: The protocol under study has been safe and effective in achieving dose-escalated radical chemoradiation in advanced cervical carcinoma. The use of fewer insertions of brachytherapy is logistically easier & can also be expected to improve compliance.

Dosimetric analysis of Deep Inspiratory Breath-hold technique (DIBH) in left-sided breast cancer radiotherapy and evaluation of pre-treatment predictors of cardiac doses for guiding patient selection for DIBH.

INTRODUCTION: The risk of radiotherapy-associated cardiovascular disease has been a concern for decades in breast cancer survivors. The objective of our study is to evaluate the dosimetric benefit of Deep Inspiratory Breath-hold technique (DIBH) on organs-at-risk (OAR) sparing in left-sided breast cancer radiotherapy and to find out pre-treatment predictors of cardiac doses for guiding patient selection for DIBH. MATERIAL AND METHODS: Pre-radiotherapy planning CT scans were done in Free Breathing (FB) and in DIBH [using Active Breathing Coordinator system (ABC™)] in 31 left sided breast cancer patients. 3DCRT plans were generated for both scans. Comparison of anatomical and dosimetric variables were done using paired t test and correlation was evaluated using Pearson correlation. Linear regression was used to get independent predictors of cardiac sparing and Receiver Operating Characteristic (ROC) curve analysis was done to find out the specific threshold of the predictors. RESULTS: There was a 39.15% reduction in mean heart dose in DIBH compared to FB (2.4 Gy vs 4.01 Gy) (p < 0.001), 19% reduction in maximum Left Anterior Descending (LAD) dose and a 9.9% reduction in ipsilateral lung mean dose (p = 0.036) with DIBH. A significant correlation was observed between reduction in Heart Volume in Field (HVIF) and Maximum Heart Depth (MHD) with reduction in mean heart dose. Reduction in HVIF (ΔHVIF) independently predicted cardiac sparing. CONCLUSION: DIBH leads to significant reduction in OAR doses and is suggested for all patients of left-sided breast cancer undergoing radiotherapy. However, HVIF and MHD predicted for cardiac sparing and threshold criteria of ΔHVIF and ΔMHD may be used by centres with high workload to select patients for DIBH.

Myeloid sarcoma: an uncommon presentation of myeloid neoplasms; a case series of 4 rare cases reported in a tertiary care institute

Myeloid sarcoma (MS) is a rare extramedullary neoplasm of myeloid cells, which can arise before, concurrently with, or following hematolymphoid malignancies. We report 04 such cases of MS, diagnosed in this institute over a period of 6 years, during various phases of their respective myeloid neoplasms/leukemias. These cases include MS occurring as a relapse of AML (Case 1), MS occurring as an initial presentation of CML (Case 2), MS occurring during ongoing chemotherapy in APML (Case 3), and MS presenting as a progression of MDS to AML (Case 4). In the absence of relevant clinical history and unemployment of appropriate immunohistochemical (IHC) studies, these cases have a high risk of being frequently misdiagnosed either as Non-Hodgkin's Lymphoma (NHL) or small round cell tumors or undifferentiated carcinomas, which may further delay their management, making an already bad prognosis worse. This case series has been designed to throw light on the varied presentation of MS and the lineage differentiation of its neoplastic cells through the application of relevant IHC markers along with their clinical correlation.

Retrospective study on performance of constancy check device in Linac beam monitoring using Statistical Process Control.

AIM: To examine the application of Statistical Process Control (SPC) and Ishikawa diagrams for retrospective evaluation of machine Quality Assurance (QA) performance in radiotherapy. BACKGROUND: SPC is a popular method for supplementing the performance of QA techniques in healthcare. This work investigates the applicability of SPC techniques and Ishikawa charts in machine QA. MATERIALS AND METHODS: SPC has been applied to recommend QA limits on the particular beam parameters using the QUICKCHECK webline QA portable constancy check device for 6 MV and 10 MV flattened photon beams from the Elekta Versa HD linear accelerator (Linac). Four machine QA parameters - beam flatness, beam symmetry along gun target direction and left-right direction, and beam quality factor (BQF) - were selected for retrospective analysis. Shewhart charts, Exponentially Weighted Moving Average (EWMA) charts and Cumulative Sum (CUSUM) charts were obtained for each parameter. The root causes for a failure in machine QA were broken down into an Ishikawa diagram enabling the user to identify the root cause of error and rectify the problem subsequently. RESULTS: Shewhart charts and EWMA charts applied could detect loss in control in one variable in the 6 MV beams and in all four variables in 10 MV beams. CUSUM charts detected offsets in the readings. The Ishikawa chart exhaustively included the possible errors that lead to loss of control. CONCLUSION: SPC is proven to be effective for detection of loss in control in machine QA. The Ishikawa chart provides the set of probable root causes of machine error useful while troubleshooting.

Rapid in-situ hybridization and immunohistochemistry: a pilot comparative study of two rapid diagnostic techniques for establishing monoclonality in plasma cell dyscrasias.

BACKGROUND: Light chain restriction needs to be established on the paraffin embedded tissue in certain types of plasma cell dyscrasias when serum levels of monoclonal immunoglobulins and light chain assays in the urine and serum may be normal. Rapid-in-situ-hybridisation (RISH) is thought to be a superior to immunohistochemistry (IHC) for kappa and lambda staining due to brighter and crisp staining without any background. METHODS: Fifty cases were included in this pilot study. Serum light chain restriction status of the case was taken as gold standard. The results of standard IHC for kappa and lambda immunoglobulins on the bone marrow biopsy of these cases was compared with RISH performed by the two commercially available kits. The results of the two methods were compared for sensitivity, need to repeat the test and background staining. RESULTS: The study found that in IHC first run sensitivity was 58% which improved to 88% after the second run. For RISH the sensitivity was 100%. CONCLUSION: Rapid-in-situ-hybridisation (RISH) is a superior technique to IHC for detecting kappa and lambda light chain in plasma cells. The test is as labour intensive and time consuming as the routine IHC but has no background staining with more bright and crisp staining quality.

Pseudoxanthomatous or xanthelasmoid mastocytosis: Reporting a rare entity.

Mastocytosis is a disease characterized by abnormal and pathologic increase in mast cells in the cutaneous tissue and extracutaneous organs such as the bone marrow, liver, spleen, lymph node and gastrointestinal tract. Cutaneous mastocytosis comprises of four major clinical variants: solitary and multiple mastocytomas, urticaria pigmentosa, diffuse cutaneous mastocytosis and telangiectasia macularis eruptiva perstans. Cutaneous mastocytosis of the xanthelasmoid type is a rare variant of diffuse mastocytosis. It is clinically characterized by the typical yellowish hue and is accompanied histologically by mast cells infiltrating far into the lower dermis. Here we report one such rare case.

Clinical Stratification of High-Grade Ovarian Serous Carcinoma Using a Panel of Six Biomarkers.

Molecular stratification of high-grade serous ovarian carcinoma (HGSC) for targeted therapy is a pertinent approach in improving prognosis of this highly heterogeneous disease. Enabling the same necessitates identification of class-specific biomarkers and their robust detection in the clinic. We have earlier resolved three discrete molecular HGSC classes associated with distinct functional behavior based on their gene expression patterns, biological networks, and pathways. An important difference revealed was that Class 1 is likely to exhibit cooperative cell migration (CCM), Class 2 undergoes epithelial to mesenchymal transition (EMT), while Class 3 is possibly capable of both modes of migration. In the present study, we define clinical stratification of HGSC tumors through the establishment of standard operating procedures for immunohistochemistry and histochemistry based detection of a panel of biomarkers including TCF21, E-cadherin, PARP1, Slug, AnnexinA2, and hyaluronan. Further development and application of scoring guidelines based on expression of this panel in cell line-derived xenografts, commercial tissue microarrays, and patient tumors led to definitive stratification of samples. Biomarker expression was observed to vary significantly between primary and metastatic tumors suggesting class switching during disease progression. Another interesting feature in the study was of enhanced CCM-marker expression in tumors following disease progression and chemotherapy. These stratification principles and the new information thus generated is the first step towards class-specific personalized therapies in the disease.

To evaluate immunoreactivity of cyclooxygenase-2 in cases of endometrial carcinoma and correlate it with expression of p53 and vascular endothelial growth factor.

CONTEXT: Increased levels of prostaglandins have been detected in cancers of several anatomic sites, including those of endometrium. Several studies have shown that cyclooxygenase-2 (COX-2) expression is aberrantly increased in various human epithelial cancers, and cellular up-regulation of COX-2 may be a common mechanism in epithelial carcinogenesis. AIMS: To examine the expressions of COX-2, p53 and vascular endothelial growth factor (VEGF) in endometrial cancer and their relationships with clinicopathologic characteristics. SETTING AND DESIGN: A retrospective observational study in a tertiary care center of academic and research potential. SUBJECTS AND METHODS: Sections from fifty cases of endometrial carcinoma were stained imunohistochemically with COX-2, p53, and VEGF. The expressions of COX-2, p53, and VEGF in endometrial cancer were examined. RESULTS: COX-2 was positive 19 cases (38%) of endometrial carcinoma. The COX-2 immunopositivity was 50%, 28%, and 41% in Grade 1, Grade 2, and Grade 3; and 27%, 46%, 67%, and 100% cases of Stage I, II, III, IV, respectively. p53 was positive in 24 cases (48%); 0%, 33%, and 67% in Grade 1, Grade 2, and Grade 3; and 27%, 77%, 83%, and 100% cases of Stage I, II, III, IV, respectively. VEGF was positive in 21 cases (42%); of which 0%, 33%, and 67% cases were in Grade 1, Grade 2, and Grade 3; and 17%, 77%, 83%, and 100% cases were in Stage I, II, III, IV, respectively. CONCLUSION: The expression of COX-2 increase with stage of the endometrial tumor and with the expression of p53 and VEGF in the endometrial carcinomas. COX-2 inhibitors may have role in the prevention of endometrial carcinomas in high-risk cases and in preventing recurrences after treatment.

Identification of Sentinel Lymph Nodes Using Contrast-Enhanced Ultrasound in Breast Cancer.

Sentinel lymph node (SLN) biopsy has become the standard of care in axillary staging of breast cancer patients who are clinically node negative as it reduces the morbidity of axillary nodal dissection. SLN biopsy using blue dye and radioisotopes have high identification rates but its limitations include anaphylaxis, disposal of radioactive waste, and potential second surgery in up to 35% of patients who show nodal metastases on SLN biopsy. Contrast-enhanced ultrasound (CEUS) has the potential for SLNs to be identified without the aforementioned risks. CEUS involves the administration of intravenous contrast agents containing microbubbles of perfluorocarbon or nitrogen gas. The bubbles greatly affect ultrasound backscatter and increase vascular contrast in a similar manner to intravenous contrast agents used in CT and MRI. It is safe and easily performed with no requirement for ionizing radiation and no risk of nephrotoxicity. Microbubbles are taken up by lymph nodes when injected directly into tissues, including sub-areolar injection in the breast cancer patient. This method may prove valuable in patients with ductal carcinoma in situ, where operative SLN biopsy remains controversial, and in women undergoing prophylactic mastectomies for high risk. This technique may also have a role after neoadjuvant chemotherapy where frequently there is fibrosis in the treated SLNs.

Role of E-cadherin and cyclin D1 as predictive markers of aggression and clonal expansion in head and neck squamous cell carcinoma.

OBJECTIVES: Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide. Inconsistency in various histopathologic features for predicting nodal metastasis and overall prognosis and a better understanding of molecular mechanisms of tumourigenesis have shifted the focus to a search for more definitive predictive markers. To identify the role of two immunohistochemical (IHC) markers, E-cadherin and cyclin D1, as predictive markers of aggressiveness in HNSCC and to assess clonal expansion of tumour cells. MATERIALS AND METHODS: A total of 66 cases of HNSCC with neck node dissection were studied. IHC was performed on primary tumour sections and lymph nodes showing metastatic deposits. Histopathological parameters such as tumour grade and TNM stage together with nodal status were compared according to expression of the two markers. Fischer's chi-square test was used to assess the correlation between the two markers and histopathological parameters. RESULTS: Out of 66 cases studied, 37 showed LN metastasis. Most of the patients were male, and the most common tumour site was buccal mucosa. We found a significant association between loss of E-cadherin and node metastasis (P<0.001) and higher TNM stage (P<0.001). Cyclin D1 overexpression was significantly associated with only nodal metastasis (P=0.007). No significant association with tumour grade was found for either marker. The subgroup of E-cadherin loss with cyclin D1 overexpression was associated with the maximum incidence of nodal metastasis and higher TNM stage, highlighting the importance of using a combination of these two markers. A significant association was noted between the expression of markers at the primary site and at nodal deposits, indicating clonal expansion. CONCLUSION: A combination of the two markers E-cadherin and cyclin D1 can predict prognosis in HNSCC, although tumour heterogeneity may affect this association in some cases.

CD44 positive/CD24 negative (stem cell like property) breast carcinoma cells as marker of tumor aggression.

BACKGROUND: Cells with stem cell like properties in solid organ malignancies like breast and pancreas have been studied over the last decade and have been found to be associated with poor prognosis. Presence of CD44 positive and CD24 negative tumor cells in breast carcinoma (cells with 'stem cell' like property) as marker of aggressiveness and poor prognosis was checked for association with various markers of disease aggression like age at presentation, size of tumor, histological grade of tumor, triple negative status, level of micro-vessel density, and nodal status. METHODS: Single and double staining immunohistochemistry protocol was used for CD24 and CD44 staining. The staining protocol was repeated with more contemporary techniques using fluorescent chromogen also. RESULTS: 52 cases, all females who underwent modified radical mastectomy at a tertiary care hospital over a period of 3 years, were evaluated. No association was found between presence of stem cells and size of lesion, histological grade, triple negative status or micro-vessel density. However, significant association was found with respect to younger age of presentation (p value = 0.044). 20 out of 25 cases with nodal metastasis were positive for presence of stem cells (p value is 0.0003). Further, 18 of these 20 cases also had stem cells in the metastatic nodule. Fluorescent chromogens (FITC and Cyanine Red) revealed similar results. CONCLUSION: Cases positive for stem cells showed earlier onset of disease and proneness to nodal metastasis.

Sclerosing angiomatoid nodular transformation of spleen masquerading as a splenic abscess.

Here we report an incidental case of sclerosing angiomatoid nodular transformation (SANT) in an open splenectomy specimen from a young man who presented with insidious abdominal pain. The condition was mimicking as a splenic abscess. Histopathological and immunohistochemical analysis confirmed this to be SANT. SANT presents a diagnostic challenge to clinicians, radiologists, as well as pathologists because of its extreme rarity. Therefore, it must be included as a differential diagnosis in cases with nonspecific pain or radiological splenic mass. This case highlights that SANT can present with varied clinical features and at a wide range of ages. A high index of suspicion is required for its diagnosis.

Cytomorphology of columnar cell variant of papillary carcinoma thyroid: A case report and review of the literature.

A 58 years old lady reported with history of progressively increasing lump in the neck. Patient had earlier undergone sub-total thyroidectomy (details not available) in a private institute one year back. Fine needle aspiration cytology (FNAC) of the present lump revealed features of papillary carcinoma thyroid. Patient subsequently underwent total thyroidectomy along with excision of a tumor nodule in the larynx. Gross examination of the specimen revealed a tumor nodule in the right lobe of the thyroid. Microscopic examination of the tumor nodule in the thyroid and larynx revealed a columnar cell variant of papillary carcinoma thyroid. Very few reports describing the cytomorphologic features of this variant of papillary carcinoma are available in the published literature. These reports highlight the absence or paucity of nuclear grooves and intranuclear inclusions in this variant. We describe a case of columnar cell variant of papillary carcinoma where nuclear grooves were prominently seen. In addition, we report the occurrence of rosette-like structures which were brought out better on FNA smears. These rosette-like structures have not been emphasized earlier in the published literature. The cytomorphologic features of this rare variant are also reviewed in this report.