Charan Singh Rayat

In Vitro Cytotoxicity of Multiwalled and Single-Walled Carbon Nanotubes on Human Cell Lines

Carbon nanotubes are the building blocks for future electronics, materials, health care devices etc. In order to explore the health care applications of nanotubes in diagnosis and drug delivery, it is important to understand their toxicity. In the present study the in vitro responses were seen when carbon nanotubes were exposed to human cell lines. It typically involves dispersion of multiwall and single walled carbon nanotubes, as well as carbon black and quartz as reference material within the cell culture medium followed by their subsequent addition to human cell lines. MTT 3-(4, 5 dimethylthiazol-2 yl) 2, 5-diphenyl tetrazolium bromide, a tetrazole assay was performed; cell viability was measured by observing their absorbance using ELISA (enzyme-linked immunosorbent assay) reader. Cell viabilities at different concentrations i.e. 50, 10, 5, 3, 1 μg mL−1 were studied. In MTT assay, it was observed that cell viability increases with decrease in concentration of single walled and multiwall carbon nanotubes. We found that both behave almost in the same manner in terms of viability in case of MTT assay. Cellular uptake of FITC (fluorescein isothiocyanate) labeled carbon nanotubes were imaged using confocal microscopy and transmission electron microscope.

Collagenofibrotic glomerulopathy—a review

Collagenofibrotic glomerulopathy (CG) is a rare cause of idiopathic nephrotic syndrome characterized by massive accumulation of atypical Type III collagen fibrils within the mesangial matrix and subendothelial space of the glomeruli. A definite diagnosis can be established when typical histological findings are supported by electron microscopy. This disease exhibits indolent progression and as yet has no specific treatment. The present article reviews the clinicopathological features, epidemiology and proposed mechanisms of pathogenesis of CG. A search of the English language literature identified 38 cases of CG, of which 22 are reported from Asian countries. An additional three cases are being reported from this Institute in India and are illustrated herein. These reports contribute to a better understanding of this disease, which although not as prevalent, should be considered as a differential diagnosis in cases of mesangiocapillary form of glomerular injury.

Correlation of Proteinuria with Podocyte Foot Process Effacement in IgA Nephropathy: An Ultrastructural Study

Abstract Background: Proteinuria is an uncommon clinical manifestation of IgA nephropathy and is usually seen in cases with severe lesions like endocapillary proliferation. However, it is occasionally seen even with cases with mild glomerular manifestations and may even be of nephrotic range. Predictor: Podocyte foot process effacement. Outcome: Severity of proteinuria. Measurements: Podocyte foot process effacement was measured. Morphometric analysis was performed on transmission electron microscope images using a computerized digital photomicrograph system (BioWizard 4.2 Image analysis software, New Delhi, India). Proteinuria was measured quantitatively assigned into five grades. Results: It was found that as the extent of proteinuria increased, the effacement ratio also increased, and this was most significant between “no” proteinuria and the rest of the categories. Conclusion: Nephrotic presentation in IgA nephropathy is a known phenomenon and in certain cases may show near normal glomerular morphology with severe foot process effacement on EM being the only significant finding to explain the proteinuria. Proteinuria in these cases shows a significant correlation with degree of foot process effacement. Renal biopsy is important in these cases because they are known to have a better prognosis and are usually steroid responsive.

Early recurrence of proliferative glomerulonephritis with monoclonal immunoglobulin deposits in a renal allograft.

Proliferative glomerulonephritis with monoclonal immunoglobulin deposits (PGNMIDs) is a clinico-pathologic entity, the recurrence of which in the renal allograft has only recently been described. A 55-year-old male presented with rapid deterioration of renal function. Light microscopy showed membranoproliferative glomerulonephritis with kappa light chain restriction and only one sub-class of IgG. He subsequently underwent renal transplant. Two months later, he developed acute graft dysfunction. Renal biopsy showed a recurrence of the disease. Work up for multiple myeloma was positive. Membranoproliferative pattern of injury in the posttransplant setting has a wide range of differential diagnosis, PGNMID being one of them.

Clinico-pathologic spectrum of C3 glomerulopathy-an Indian experience

BackgroundC3 glomerulopathy (C3GP) is characterized by deposition of complement C3 with absence/traces of immunoglobulins in the glomeruli and categorized into dense deposit disease (DDD), C3 glomerulonephritis (C3GN), complement factor H related protein 5(CFHR5) nephropathy etc. Collaborative efforts of pathologists, complement biologists and nephrologists worldwide are expanding the histomorphological pattern and laboratory findings related to C3GP. Hence, we studied point prevalence and morphological spectrum of C3GP in Indian patients to correlate morphological patterns with standard therapies and outcome of the patients.MethodsRetrospective analysis of renal biopsies (2007-2012,n-4565), which on immunofluorescence (IF) had C3 dominant deposits with absence or trace amount of immunoglobulin was carried out. Histopathology and electronmicroscopy (EM) were reviewed; cases were re-classified as DDD and C3GN. Histomorphological patterns of both groups were compared and correlated with treatment. Clinical details and follow up of patients were retrieved from the department of nephrology.ResultsThere were 31 cases (0.7%) of C3GP sub-classified as DDD (n-13) and C3GN (n-14). It was difficult to sub-classify 4 cases since EM showed overlapping features. C3GN and DDD had distinct clinical characteristics and disease outcome, though pathological features were overlapping. Majority of C3GP patients were males and were in 2nd to 4th decade of life. Nephrotic syndrome in DDD and nephritic-nephrotic presentation in C3GN patients was more common.Hypertension and oliguria were more often observed in C3GN than DDD. Membranoproliferative pattern (MPGN) was commonest pattern in DDD; other patterns seen were mesangial proliferative, mesangial expansive/nodular, exudative and crescentic. C3GN also had all the above patterns, the predominant ones being MPGN and mesangial proliferative. Limited follow-up revealed response to therapy only in C3GN (33%). Progression to ESRD was 33% in DDD and 10% cases in C3GN.ConclusionC3GP comprise 0.7% of all renal biopsies. MPGN pattern was the commonest morphological pattern in DDD whereas MPGN and mesangial proliferative pattern were equally dominant patterns in C3GN. EM of 4 cases (13%) showed intermediate features. Evaluation of alternate complement pathway must be done in all cases to identify the point of dysregulated alternate complement pathway and to confirm the diagnosis in ambiguous cases.Virtual slidesThe virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1730070964135632

Spectrum of pulmonary adenocarcinoma with ultrastructural correlation: An autopsy study from northern India

BACKGROUND In the present study, we have evaluated the use of electron microscopy in subtyping pulmonary adenocarcinomas, comparing the ultrastructural findings with the diagnosis rendered by light microscopy. MATERIALS AND METHODS The gross and histologic features of 16 autopsy cases of pulmonary adenocarcinoma were analyzed and compared with electron microscopic features. The cytologic phenotypes of these cases of well-differentiated pulmonary adenocarcinoma were determined by electron microscopic examination. More than 200 cells in each case were examined, and the tumors were classified according to the predominant feature noted. RESULTS Eight cases were of Clara cell origin and one case each of type II pneumocyte and bronchial surface cell type. The remaining 6 cases lacked definite discernible features of differentiation towards any specific cell type, other than presence of small nuclear clefts in occasional nuclei. Tumors with Clara cell differentiation were low cuboidal with apical snouts. Type II pneumocyte tumor failed to reveal any characteristic definable as light microscopic feature. CONCLUSION Ultrastructural examination is the only definite means of identification of various cell types in the respiratory epithelium and hence forms an invaluable tool in classification of pulmonary adenocarcinoma.

Podocin and Beta Dystroglycan expression to study Podocyte-Podocyte and basement membrane matrix connections in adult protienuric states

BackgroundPodocytes can be the primary site of injury or secondarily involved in various protienuric states. Cross talk between adjacent foot processes and with basement membrane is important for slit diaphragm function. Does expression of podocyte associated proteins in kidney biopsies alter with site/type of primary injury? Genetic mutations of podocin result in steroid resistant FSGS. Can protein expression of podocin predict resistant cases to initiate further genetic evaluation?MethodsAdult patients (n-88) with protienuria- minimal change disease(MCD)-22, focal segmental glomerulosclerosis(FSGS)-21,membranous glomerulonephritis(MGN)-25 and IgA nephropathy(IgAN)-20 were selected for immunohistochemistry with podocin and beta dystroglycan . Results were graded (0 - 3+scale )and compared with control biopsies and internal control. Treatment and follow up (6 months -2 ½ years) of FSGS and MCD cases were collected.ResultsThere was intense to moderate staining of the podocytes with podocin and β dystroglycan in the glomeruli in all cases (MCD, FSGS, IgAN and MGN) except for weak staining with β dystroglycan in 3 cases of MCD. There was loss of immunostains in areas of segmental/global sclerosis. There was no significant difference in the staining pattern between the groups. In primary podocytopathies, staining pattern did not differ between steroid resistant, sensitive or dependent cases.ConclusionsImmunohistochemical expression of podocin and β dystroglycan does not differ in nephropathies which have different site of injury depending on absence (MCD and FSGS) or presence of immune deposits and their localization (MGN and IgAN). Podocin and β dystroglycan staining did not differentiate steroid sensitive and resistant cases, hence, does not give clue to initiate genetic studies. However, analysis of bigger cohort may be required.SummaryPodocin and β dystroglycan immunohistochemistry was done to analyze podocyte - podocyte and podocyte -basement membrane matrix connections in adult protienuric states. Primary podocytopathies i.e. MCD and FSGS and secondary podocytopathy due to immune complex deposition i.e. MGN (subepithelial) and IgAN (mesangial) were analyzed. There was no difference in staining patterns between primary and secondary podocytopathies or between steroid sensitive, resistant and dependent cases of FSGS and MCD.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2258608781052786

Acute renal failure due to Dengue myositis: a rare cause of pigment cast nephropathy

Dengue is a common viral infection of the tropics caused by four serotypes of Dengue virus belonging to the family of flavivirus. Clinical presentation of symptomatic Dengue virus infections can be wide, from a mild febrile illness to a life-threatening shock syndrome. We present a case of a young male with unusual presentation of Dengue as acute renal failure secondary to rhabdomyolysis.

Pigmented intraventricular tumor in a young adult: Pigmented central neurocytoma

A 22-year-old man presented with complaints of progressively worsening headache for the past 6 months. Clinical examination revealed papilledema. There were no sensory or motor deficits. Radiology showed an intraventricular heterogeneous lesion occupying mainly the body of right lateral ventricle with some growth into the left lateral ventricle (Fig. 1). The features suggested central neurocytoma. He was operated through an endoscopic approach that required conversion to open surgery due to intraoperative bleeding. The tumor was fleshy and vascular. Although near total excision was achieved, he succumbed to ventriculitis.