Formulation Development and Evaluation of Paclitaxel Loaded Solid Lipid Nanoparticles Using Glyceryl Monostearate.

Solid Lipid Nanoparticles (SLNs) are important because of their size and stability. SLNs have been reported as an alternative drug delivery device to traditional polymeric nanoparticles. SLNs are in submicron range (50- 1000nm) and are composed of physiologically tolerated lipid components. At room temperature the particles are in solid state. They are made up of bio-compatible and bio-degradable materials capable of incorporating lipophilic and hydrophilic drugs. Paclitaxel is a Di-terpenoid Pseudo-alkaloid having anti-neoplastic activity particularly against primary epithelial, ovarian carcinoma, Breast cancer, Colon Cancer, Brain Cancer, Lungs cancer and AIDs Related Kaposi’s Sarcoma. Paclitaxel is an effective drug against Aggressive Cancer’s because it adversely affect the process of cell division by preventing restructuring. The present study is to investigate the probability of incorporating paclitaxel in SLNs using Glyceryl Mono-stearate (GMS) as a lipid matrix, poly-oxy ethylene (Brij 97) as a surfactant, soya-lecithin as a co-emulsifier. Paclitaxel loaded SLNs are prepared by Solvent emulsification and evaporation method using ultra sonication and optimization of critical process variables were carried out to develop stable SLNs. The average particle size of SLNs was found to be 63nm ± 5.77 with Poly dispersity index (PDI) 0.272 ± 0.02 and entrapment efficiency was found 94.58%. The stability studies and zeta potential were performed at refrigerated temperature (2-8 OC) indicating no significant increase in particle size after one month storage. In-vitro release studies showed initial burst release followed by controlled release for 48hrs (about 73%). The release profile was fitted into Higuchi’s model (r2=0.9774). The drug diffuses from SLNs at a comparatively slower rate as the distance for diffusion increases.

Diagnostic dilemma in histopathology report following robot assisted laparoscopic prostatectomy: Tumour 'hide and seek'.

INTRODUCTION: Widespread PSA (prostate specific antigen) screening has resulted in stage migration of prostate cancer. Smaller tumor volumes are being detected in radical prostatectomy specimens. This has coincided with increasing reports about the ‘vanishing cancer phenomenon.’ AIMS: To analyse the cases of robot assisted laparoscopic prostatectomy (RALP) at our institute in which the pre operative prostate biopsy was positive for adenocarcinoma but no tumor could be identified in the final histopathology, and to review the literature for possible reasons for such a phenomenon. MATERIALS AND METHODS: Nine patients were identified out of a total of 184 cases of RALP in which the final histopathology did not correlate with the initial biopsy report. The initial biopsy slides as well as the final histopathology slides were reviewed by a second pathologist. The specimens were processed in entirety and additional sections were taken until no tissue was left. RESULTS: Two patients had cancer diagnosed on TURP (transurethral resection of prostate) chips, while the remaining patients had undergone TRUS biopsy for elevated PSA. The final histopathological diagnosis was benign prostatic hyperplasia in two patients, chronic prostatitis in four patients, and acute florid prostatitis in one patient, granulomatous prostatitis with glandulostromal hyperplasia in one patient and TCC (transitional cell carcinoma) of prostate in one patient. CONCLUSION: Most cases of pT0 are due to inability of routine histopathological analysis to identify minute tumor focus. Urologists need to be aware of this in view of the potential medico legal implications.

HIV associated renal disease: A pilot study from north India.

Background & objectives: HIV/AIDS patients may have renal involvement also, however, Indian data are sparse. The present study was done to find the spectrum of renal diseases in HIV/AIDS patients in north India. Methods: In this prospective pilot study, HIV positive patients aged >18 yr were screened for renal involvement [serum creatinine >1.5 mg% and/or significant proteinuria (>500 mg /day)]. Patients who were positive on screening were followed up prospectively and underwent kidney biopsy if indicated. Results: A total of 526 patients were screened, of these, 91 (17.3%) were found to have renal involvement. Group A (Treatment naïve) comprised 392 patients who were not on antiretroviral treatment (ART) and group B (patients on ART) comprised 134 patients. More patients (74/392, 18.9%) in group A had renal involvement as compared to patients in group B (17/134, 12.7%). Of the 91 patients with renal involvement, 26 were followed up and underwent kidney biopsy. Thirteen patients had only proteinuria and another 13 had renal dysfunction with or without proteinuria. Most common histological diagnosis was mesangioproliferative glomerulonephritis (mes PGN) (10/26). Two patients had collapsing FSGS (focal segmental glomerulosclerosis) and three patients had immune complex glomerulonephritis. Seven patients had acute kidney injury, whom six totally recovered from their renal function. All patients with mesPGN tolerated angiotensin converting enzyme (ACE) inhibitors well. There was mixed response of collapsing FSGS to steroids. Both patients with MPGN (membranoproliferative glomerulonephritis) did well on low dose of steroid and ART. Interpretation & conclusions: Renal involvement was found to be common in HIV positive patients (17.3%). A low occurrence of renal involvement found in patients already on ART suggests some renoprotective effect of ART. Our preliminary results showed that collapsing FSGS was not rare in Indian HIV positive population, but classical HIV associated nephropathy was not seen. Longitudinal studies with robust study design and large sample size need to be done to confirm the findings.

Prevalence of renal involvement in Indian patients with systemic sclerosis.

BACKGROUND: Renal involvement in systemic sclerosis (SSc) either in the form of scleroderma renal crisis (SRC) or nonrenal crisis abnormalities has been reported to be in the range of 60-80%. Renal involvement is thought to be rare in Indian patients with SSc. However, there is paucity of data. AIMS: To study the frequency and pattern of renal involvement in Indian patients with SSc. SETTINGS AND DESIGN: A single center prospective, cross sectional study. MATERIALS AND METHODS: We prospectively evaluated the patients with SSc attending the Rheumatology Clinic. All patients were evaluated for renal involvement. All patients underwent measurement of blood pressure, urine examination, glomerular filtration rate (GFR) estimation using Cockcroft-Gault formula and kidney biopsy when indicated. STATISTICAL ANALYSIS: Statistical analysis was performed using SAS 8.0 statistical package. RESULTS: Eighty-seven patients were included in the study from July 2001 to December 2004. Mean age of patients was 36.88 +/- 12.51 years. About 30% of patients had diffuse cutaneous SSc. None of these patients had SRC either at enrollment in the study or during follow-up. Eleven (12.6%) patients had hypertension. Six (6.9%) patients had abnormal urinary findings in the form of either active urinary sediment or significant proteinuria. Only one patient had azotemia (plasma creatinine > 1.8 mg/dl). Calculated GFR CONCLUSION: SRC is very rare in Indian patients with SSc. However, non-renal crisis abnormalities appear to be as common in Indian patients as compared to the western literature.

Frequency and role of hepatitis-C virus and type II cryoglobulinemia in membranoproliferative glomerulonephritis.

BACKGROUND: Many studies have claimed a major role of chronic hepatitis-C virus (HCV) infection in immune-mediated diseases such as membranoproliferative glomerulonephritis (MPGN). Chronic HCV infection is also known to produce essential mixed cryoglobulinemia (EMC), which in turn may manifest as vasculitis and cryoglobulinemic MPGN. OBJECTIVE: The aim of the study therefore, was to determine frequency of association and pathogenetic role of HCV infection as well as that of EMC in MPGN patients. METHODS: Fifty-three adult patients of MPGN were studied for HCV, HBsAg, EMC, C3, anti-nuclear antibody (ANA), rheumatoid factor serologically. Histopathology, immunofluorescence (IF) were conducted in all patients and electron microscopy (EM) in those who were found HCV positive. Simultaneously 37 follow-up patients of HCV associated chronic hepatitis were investigated for EMC, renal functions and urinalysis done for evidence of glomerulonephritis (GN). RESULTS: Thirteen percent MPGN patients were HCV positive, however, no viral particle could be seen in electron microscopy in glomeruli of these patients. There was no serologic evidence of HCV induced immune complex GN. None of the MPGN patients showed cryoglobulinaemia. Similarly none from HCV associated chronic hepatitis group had EMC nor showed evidences of glumerulonephritis. CONCLUSION: Thirteen percent of adult MPGN patients in north India were seropositive for HCV, indicating significant association. However, clear evidence in favour of its pathogenetic role was lacking in our study. Secondly, this study reveals that MPGN is non-cryoglobulinemic and HCV is not a major cause in our population compared to what is reported from other countries. These observations need confirmation by a larger study.

Comparative evaluation of grading of breast carcinomas in fine needle aspirates by two methods.

BACKGROUND & OBJECTIVES: Fine needle aspiration (FNA) material is increasingly being used to determine various prognostic parameters in patients with carcinoma breast. It was recommended that cytological grading should be part of all FNA reports of breast carcinoma so that preoperative prognostication could be done on it. However, the most reliable method for cytological grading that closely reflects the most widely used histological grading system is yet to be determined. This study was undertaken to compare results of two cytological grading methods to see which corresponded better to the histological grade. METHODS: In a double-blind study, cytological grading of 52 patients with breast carcinoma was carried out using two different grading systems (Robinson's and Mouriquand's methods) and the grades compared with the histological grading to find out which reflected histological grades more closely. RESULTS: By Robinson's method 28.8, 46.2 and 25 per cent aspirates were graded as I, II and III respectively. Using Mouriquand's grading 9.6, 69.2 and 21.2 per cent aspirates were graded correspondingly. Comparison of the two methods showed a concordance in 40 (76.9%) cases. Both cytological grading systems showed a concordance of 71.2 per cent with the histological grading. Robinson's cytological grading showed a diagnostic accuracy of 80.76 per cent with 77.77 per cent specificity while Mouriquand's method had an accuracy of 84.60 per cent with 33.33 per cent specificity. INTERPRETATION & CONCLUSION: Though the Robinson's and Mouriquand's grading systems were found to have similar concordance with histological grading, Robinson's method was considered better because of its simplicity, and specificity.

An Asymptomatic Young Female with Chronic Hepatitis-B Presenting as Minimal Change Glomerulonephritis with Nephrotic Syndrome

Hepatitis B is a worldwide problem with high prevalence rate in our country. Kidney involvement is common in chronic hepatitis B patients. The usual pattern is membranous glomerulonephritis in children and Type I membranoproliferative glomerulonephritis in adults. A previously asymptomatic young female suffering from chronic hepatitis B with portal hypertension presented with nephrotic syndrome. Kidney biopsy revealed minimal change glomerulonephritis which is rarely seen in association with hepatitis B. Patient recovered following administration of oral steroids. We reconunend screening of all patients of nephrotic syndrome for chronic hepatitis B with viral markers in addition to HBsAg.

Pulmonary mucormycosis in a diabetic renal transplant patient.

A 50-year-old male, diabetic, post transplant patient had recurrent pneumonia. The first episode responded well to antibiotics, while on the second occasion he had a necrotising pneumonia, which developed into a thick-walled cavity. Despite antibiotics, his condition rapidly deteriorated precluding bronchoscopy or percutaneous biopsy. Post-mortem lung biopsy revealed typical hyphae of mucormycosis.

Hepatitis C virus related cryoglobulinemic glomerulonephritis.

A 46 years male presented with skin rash and acute nephritic syndrome. He had history of jaundice four months back. Rheumatoid factor and cryoglobulins were present in the serum. Although anti-HCV antibodies were negative, HCV RNA was detected by polymerase chain reaction. Kidney biopsy showed membranoproliferative glomerulonephritis and thrombi in the lumen of the glomerular capillary loops. His renal functions improved with steroids.

Embryonal sarcoma of the liver mimicking a hydatid cyst in an adult.

Undifferentiated (embryonal) sarcoma of the Liver is a rare malignant mesenchymal tumour. This presents predominantly in late childhood. It carries a dismal prognosis. Surgical resection with or without adjuvant therapy remains the mainstay of treatment. The present report describes a sixteen year old girl who presented with a large hepatic mass mimicking a hydatid cyst: clinically, radiologically and on serology.

A study of autoimmune markers in hepatitis C infection.

BACKGROUND & OBJECTIVES: Hepatitis C virus (HCV) infection is associated with several autoimmune markers. Despite HCV being common in India, no information on this aspect is available. This study was undertaken to ascertain the frequency and clinical significance of autoimmune markers like rheumatoid factor (RF), antinuclear antibodies (ANA), antibodies to double stranded deoxyribonucleic acid (dsDNA), anti neutrophil cytoplasmic antibody (ANCA), anti smooth muscle antibodies (ASMA), anti liver kidney microsomal 1 antibodies (anti LKM1), anti gastric parietal cell antibodies (anti GPCA), anti mitochondrial antibodies (AMA), anti cardiolipin antibodies (ACL) and cryoglobulins in HCV infection and to determine the effect of treatment on these markers. METHODS: Twenty five patients with chronic hepatitis C and 25 healthy controls were studied. Cryoglobulins were detected by cryoprecipitation, RF by latex agglutination, anti dsDNA and ACL by ELISA while indirect immunofluorescence was used to detect all other autoantibodies. RESULTS: Eighteen patients (72%) demonstrated autoimmune markers. RF, cryoglobulins and anti LKM1 antibodies were the most frequently detected markers (in 32% patients each). ASMA, perinuclear ANCA (pANCA), ANA and anti GPCA were seen in 24, 20, 12 and 4 per cent patients respectively. None of the patients exhibited ACL, AMA or antibodies to dsDNA. No antibodies were detected in healthy controls. Sixty per cent of the patients had rheumatological symptoms. Of the seven patients followed up after treatment with alpha interferon, only two exhibited persistence of RF, while symptoms and other markers disappeared. INTERPRETATION & CONCLUSION: Rheumatological symptoms and autoimmune markers are common in HCV infection and are usually overlooked. Patients with unexplained joint pains and/or palpable purpura should be screened for HCV. Further studies are needed to delineate fully the link between infection and autoimmunity.

Classical polyarteritis nodosa and microscopic polyangiitis--a clinicopathologic study.

OBJECTIVE: To describe the clinical spectrum, laboratory features, histopathological findings and treatment outcome in patients with classical polyarteritis nodosa (PAN) and microscopic polyangiitis (MPA). MATERIAL AND METHODS: Patients with PAN and MPA seen at a large teaching hospital in north India over a period of five years (1994-99) were included in the present study. RESULTS: We encountered five patients with PAN and six patients with MPA during the study period. Of the five patients with PAN, two had systemic disease while three had limited PAN. The patients with limited PAN included two with cutaneous PAN and one with PAN confined to the nerves. Constitutional symptoms, musculoskeletal complaints, peripheral neuropathy and skin lesions dominated the clinical picture. Fifty percent of the MPA patients presented as pulmonary renal syndrome. All the patients with PAN were HBsAg and ANCA negative and had normal urinalysis findings. In contrast, all patients with MPA demonstrated an active urine sediment and 83.3% were pANCA positive. Some of the rare features encountered by us were the presence of antiphospholipid syndrome and extensive interstitial lung disease in MPA, and spontaneous recovery in one patient with systemic PAN. Treatment outcome was better in PAN as compared with MPA. CONCLUSIONS: The clinical spectrum of PAN and MPA is quite varied. A good outcome is possible with the use of corticosteroids and cyclophosphamide.

Use of complement receptor 1 (CD35) assay in the diagnosis and prognosis of immune complex mediated glomerulopathies.

Complement Receptor 1 (CR1) is a polymorphic glycoprotein expressed on erythrocytes, leukocytes and glomerular podocytes and has a major role in immune complex processing. In addition, it regulates the complement cascade activation by preventing formation of classical and alternative pathway convertases and by acting as a cofactor for Factor I mediated cleavage of C3. In this study, we have examined the expression of erythrocyte CR1 (E-CR1) and glomerular CR1 (G-CR1) in different kinds of nephropathies using ELISA and immunofluorescence microscopy to understand their role in immune complex (IC) mediated renal diseases. E-CR1 was significantly reduced in all categories of lupus nephritis in comparison to normal subjects and non-IC renal diseases. However, other IC mediated diseases like IgA nephropathy and membranoproliferative glomerulonephritis had normal E-CR1 levels. G-CR1 showed distinct differences between IC and non-IC mediated diseases. G-CR1 was virtually absent in lupus kidneys. In other IC mediated diseases, there was a correlation of G-CR1 expression to the IC and complement fragment deposition. G-CR1 serves as a useful diagnostic marker for IC mediated diseases while E-CR1 is useful as a prognostic marker to monitor the course of disease after the treatment has initiated.

Imagecytometry: a new tool for diagnosis of glomerular haematuria.

Differentiation between glomerular and nonglomerular haematuria is a major challenge in clinical medicine, which is very important for a definitive diagnosis and management in individual cases. Phase contrast microscopy of red cells in urine is the standard practice for diagnosis of glomerular haematuria. Urine cell flowcytometry is recently being used for such diagnosis. In this context, the role of determination of haemoglobin content of urine red cells is not know. Application of image analysis to study the red cells in urine may be more objective and accurate for the diagnosis. The present study has been undertaken to evaluate the urine red cells with the help of an automated computerized image analysis system for determination of hemoglobin content by integrated optical density (IOD). The morphometric parameters were also analyzed. The glomerular RBCs were significantly smaller in diameter, area and perimeter than nonglomerular RBCs with a greater variation in shape and lower [OD (p<0.0001 to <0.00002). With the help of morphometric parameters the percentage of cases diagnosed correctly varied from 90 to 95. The IOD helped to diagnose 100% cases. Thus application of this new technique may be very useful diagnostic tool in the investigation of haematuria.