Role of rapid on-site evaluation in CT-guided fine needle aspiration cytology of lung nodules

Objective: To prospectively investigate the value of rapid on-site evaluation (ROSE) in transthoracic fine needle aspiration cytology (FNAC) of patients with pulmonary nodules. Computed tomography (CT)-guided FNA is commonly employed for the diagnosis of lung lesions and the most common reason for not being able to provide a diagnosis in FNA is inadequacy of samples. Materials and Methods: This was a prospective study conducted in the departments of pathology and radiology of our cancer centre. This study had approval from the institutional review board and ethical committee of our institute. Fifty consecutive cases undergoing CT-guided transthoracic FNAC in our centre were included in the study. The smear submitted for ROSE was stained using toluidine blue stain. The specimen adequacy and diagnosis in ROSE was compared with that of the final cytology report, and the concordance regarding adequacy and diagnosis were noted. Results: Smears were adequate in 34 cases (68%) and inadequate in 16 cases (32%) Out of the 16 inadequate cases, 5 (31%) were converted to adequate due to the application of ROSE, thus increasing the adequate number of cases to 39 (78%). A diagnosis of malignancy was made in all 39 adequate cases. Sensitivity of ROSE in determining adequacy was 92% and specificity was 100%. The most common malignancy was adenocarcinoma in 26 cases. Pnemothorax occurred in 2 cases. No significant complications occurred in other cases. Conclusion: CT-guided FNA with ROSE is a safe and useful tool in the diagnostic work-up of lung cancer patients. A multidisciplinary approach along with onsite evaluation of adequacy will increase the diagnostic utility of cytology in lung lesions

Curcumin Loaded-PLGA Nanoparticles Conjugated with Tet-1 Peptide for Potential Use in Alzheimer’s Disease

Alzheimer’s disease is a growing concern in the modern world. As the currently available medications are not very promising, there is an increased need for the fabrication of newer drugs. Curcumin is a plant derived compound which has potential activities beneficial for the treatment of Alzheimer’s disease. Anti-amyloid activity and anti-oxidant activity of curcumin is highly beneficial for the treatment of Alzheimer’s disease. The insolubility of curcumin in water restricts its use to a great extend, which can be overcome by the synthesis of curcumin nanoparticles. In our work, we have successfully synthesized water-soluble PLGA coated- curcumin nanoparticles and characterized it using different techniques. As drug targeting to diseases of cerebral origin are difficult due to the stringency of blood-brain barrier, we have coupled the nanoparticle with Tet-1 peptide, which has the affinity to neurons and possess retrograde transportation properties. Our results suggest that curcumin encapsulated-PLGA nanoparticles are able to destroy amyloid aggregates, exhibit antioxidative property and are non-cytotoxic. The encapsulation of the curcumin in PLGA does not destroy its inherent properties and so, the PLGA-curcumin nanoparticles can be used as a drug with multiple functions in treating Alzheimer’

Solid-pseudopapillary neoplasm of the pancreas: A classical presentation with unique paranuclear dot like immunostaining with CD 99

A 32-year-old lady presented with a history of abdominal pain and upper abdominal discomfort of 3 months duration. Her imaging studies done at a local hospital showed a solid-cystic mass involving head of the pancreas. The patient was referred to our surgical oncology department. On examination, there was a nontender mass in the epigastrium. An ultrasound scan guided fine-needle aspiration (FNA) was done which was showing classical features of solid-pseudo papillary neoplasm of the pancreas. With this preoperative diagnosis patient was taken up for surgery. Per operatively, there was a solid-cystic mass in the head of the pancreas. Pancreaticoduodenectomy was done. Histopathology and immunohistochemistry (IHC) confirmed the diagnosis of solid-pseudo papillary neoplasm of the pancreas. Apart from the routine IHC panel, CD 99 immunostain was also done which demonstrated the characteristic paranuclear dot-like staining observed in previous studies in the literature

Proteomic profiling of high glucose primed monocytes identifies cyclophilin A as a potential secretory marker of inflammation in type 2 diabetes

Hyperglycemia is widely recognized to be a potent stimulator of monocyte activity, which is a crucial event in the pathogenesis of atherosclerosis. We analyzed the monocyte proteome for potential markers that would enhance the ability to screen for early inflammatory status in Type 2 diabetes mellitus (T2DM), using proteomic technologies. Monocytic cells (THP-1) were primed with high glucose (HG), their protein profiles were analyzed using 2DE and the downregulated differentially expressed spots were identified using MALDI TOF/MS. We selected five proteins that were secretory in function with the help of bioinformatic programs. A predominantly downregulated protein identified as cyclophilin A (sequence coverage 98%) was further validated by immunoblotting experiments. The cellular mRNA levels of cyclophilin A in various HG-primed cells were studied using qRT-PCR assays and it was observed to decrease in a dose-dependent manner. LC-ESI-MS was used to identify this protein in the conditioned media of HG-primed cells and confirmed by Western blotting as well as ELISA. Cyclophilin A was also detected in the plasma of patients with diabetes. We conclude that cyclophilin A is secreted by monocytes in response to HG. Given the paracrine and autocrine actions of cyclophilin A, the secreted immunophilin could be significant for progression of atherosclerosis in type 2 diabetes. Our study also provides evidence that analysis of monocyte secretome is a viable strategy for identifying candidate plasma markers in diabetes

TGA Analysis of PLGA nanoparticle, raw curcumin and PLGA-curcumin nanoparticle.

<p>Curcumin undergoes a gradual thermal decomposition when compared to the rapid decomposition in PLGA and PLGA-curcumin nanoparticles.</p

Flow cytometry analysis of the uptake of the targeted and non-targeted nanoparticles based on the fluorescent intensity of the nanoparticle uptake in GI-1 glioma cells.

<p>A- uptake of curcumin-PLGA nanoparticles; B- uptake of Tet-1 conjugated curcumin-PLGA nanoparticles. Enhanced uptake of Tet-1 attached nanoparticles were observed when compared to the curcumin-PLGA nanoparticles (without Tet-1).</p

Cytotoxicity analyses results by Alamar Blue assay (A) and MTT assay (B) after 24 hour incubation with the nanoparticle.

<p>Three different concentrations of the test samples were added to the cells and incubated for 24 hours before adding the respective assay reagents. We have observed that the nanoparticles were highly biocompatible.</p

Confocal imaging of nanoparticle uptake by GI-1 cells.

<p>A, B, C- PLGA-curcumin nanoparticles (without Tet-1 peptide) are easily taken up by the cell and are found distributed all though the cell cytoplasm. D, E, F- Nanoparticles targeted with Tet-1 peptide show greater affinity towards the cell soma and nucleus. A& D- Bright-field image, B& E- stained with DAPI nuclear stain observed in blue, C&F- autofluorescent PLGA-curcumin nanoparticles are observed in green. The scale bars on all the images correspond to 20 µm.</p

Anti-amyloid activity of PLGA-curcumin nanoparticles.

<p>A- Amyloid protein (Aβ) aggregates, B- Aβ and PLGA-curcumin nanoparticles after 12 hour co-incubation, C- Aβ and PLGA-curcumin nanoparticles after 24 hour co-incubation, D- Aβ and PLGA-curcumin nanoparticles after 48 hour co-incubation, E- Tet-1 conjugated PLGA-curcumin nanoparticles, F- Aβ and Tet-1 attached PLGA-curcumin nanoparticles after 12 hour co-incubation, G- Aβ and Tet-1 attached PLGA-curcumin nanoparticles after 24 hour co-incubation, H- Aβ and Tet-1 attached PLGA-curcumin nanoparticles after 48 hour co-incubation. It is observed that the nanoparticles are able to attach with the amyloid aggregates and help in disaggregation of the amyloid protein aggregates.</p