A Rare Case of Endolymphatic Sac Tumour: Clinical, Radiological, Pathological and Immunohistochemical Findings with Review of Literature.

Endolymphatic sac tumour (ELST) is a rare low grade malignant epithelial tumour of the petrous temporal bone, thought to arise from papillary epithelium of the endolymphatic sac. They may occur sporadically or in association with Von-Hippel Lindau disease. ELST is extremely rare neoplasm with benign histopathological appearance and clinically destructive behaviour. Because of the rarity of this tumour, it can easily be confused with other tumours such as paraganglioma, middle ear adenoma, metastatic carcinomas or choroid plexus papilloma. We report here a rare case of ELST with review of literature and discuss the differentiating features of ELST from its mimickers, showing a papillary configuration.

Clinico-radiological-pathological correlation in eumycetoma spectrum: Case series.

Maduramycosis or mycetoma is one of the chronic granulomatous diseases commonly seen in tropical regions. Microbiological cultures and biopsy are carried out for the definitive diagnosis of the disease, but they are time-consuming methods. The present study aims to correlate clinical, radiological and pathological features in eumycetoma cases to emphasize the role of "dot in circle" sign leading to early imaging-based diagnosis. Imaging plays an important role in early diagnosis of mycetoma, which has therapeutic implications. "Dot in circle" sign is a recently described sign in mycetoma cases on ultrasound (USG) and magnetic resonance imaging (MRI). We diagnosed four cases of eumycetoma based on clinical and imaging features, which were confirmed with histopathology. The differential diagnosis, which may mimic this sign is also discussed. The "dot in circle" sign is seen on both ultrasound and MRI. This sign is highly specific for mycetomas. Knowledge of this sign can help in early diagnosis of mycetomas preventing misdiagnosis and further complications.

Toxic Leukoencephalopathy: An unusual Presentation by 5-Fluorouracil Infusion.

BACKGROUND: Toxic leukoencephalopathy predominantly affect white matter of the brain parenchyma. Patient presents either in acute, subacute or chronic phase. The clinical presentation may vary, ranging from mild cognitive impairment to severe neurological dysfunction. It can also mimic psychiatric illness. CASE REPORT: A 50-year-old woman was diagnosed with locally advanced carcinoma buccal mucosa. She was planned for Neoadjuvant chemotherapy consisting of Docetaxel, Cisplatin, 5-FU (TPF). During 3rd day of 3rd cycle of 5-fluorouracil (5FU) infusion, patient developed hypoactive delirium and later became comatose state of drowsiness, with Glasgow Coma Scale (GCS) was 5. There was no previous history for the same. Hence, the infusion was stopped. Patient was evaluated with NCCT head. No abnormality was seen on CT scan. MRI brain was done and it showed diffusion restriction with T2 / FLAIR hyper intensities in bilateral centrum semiovale, white matter of bilateral parietal region and in corpus callosum. Patient received symptomatic care, nutrition by ryles tube during this period and she started to improve after 1st week of onset of symptoms. After 3 weeks, MRI was repeated and there was complete resolution of previous findings. CONCLUSIONS: Development of neurological symptoms during 5FU infusion is a rare entity. Henceforth, occurrence of toxic leukoencephalopathy should be kept in mind. Diffusion weighted imaging play an important role in both acute episode of toxic encephalopathy and in follow up.

Potent Acetylcholinesterase Selective and Reversible Homodimeric Agent Based on Tacrine for Theranostics.

Acetylcholinesterase (AChE) has been an important biomarker for diagnosing Alzheimer's disease (AD), due to reduction in AChE activity in post-mortem brains of AD patients. A potent, selective, and reversible homodimeric inhibitor of AChE, 5-amino- N1, N3-bis(2-(1,2,3,4-tetrahydroacridin-9-ylamino)ethyl)isophthalamide (compound 4), was synthesized by using 9-alkyl(1,2,3,4-tetrahydroacridine) pharmacophore with appended functionality. In the present work, we report the synthesis of this bivalent inhibitor of AChE. The homodimeric ligand structure was designed and studied with molecular docking tools, which revealed its high affinity and interactions with active site gorge of AChE, which includes both catalytic active site (CAS) and peripheral active site (PAS). The IC50 value of this bivalent inhibitor for AChE and BuChE were 0.54 ± 0.06 and 32.49 ± 1.2 nM, respectively, with a selectivity ratio of 60.16 toward AChE. The designed ligand also showed potent inhibitory properties on PAS activity as well as on AChE-induced amyloid aggregation with low cytotoxicity on rat hippocampal neurons. The AFM images further corroborated the Aß1-42 aggregation inhibition by compound 4 to an extent similar to bis(7)-tacrine. Moreover, the bivalent ligand was also proven to be of neurogenic potential due to its ability to induce S-phase post-treatment in rat hippocampal neuronal cells. On the basis of initial results, the agent could be further explored for its theranostic value clinically, which gives the possibility of tracing the AChE levels by molecular imaging techniques in correlation with progression of neurocognitive disorders like AD for better therapy response and patient management.

Radiolabeling and Preclinical Evaluation of a New S-Alkylated Cysteine Derivative Conjugated to C-Substituted Macrocycle for Positron Emission Tomography.

A new S-alkylated cysteine-derivatized tumor targeting agent, 2,2'-(12-(2-((2-acetamido-2-carboxyethyl)thio)acetamido)-11,13-dioxo-1,4,7,10-tetraazacyclotridecane-4,7-diyl)diacetic acid was developed for positron emission tomography (PET) imaging. N-Acetyl cysteine (NAC) was conjugated to ATRIDAT as a specific targeting agent toward L-type and ASC amino acid transporter systems in the oncogenic cells. NAC was attached via S-alkylation to prevent its incorporation at undesired recognition sites affecting the signal-to-noise ratio. NAC-ATRIDAT was subjected to gallium-68 complexation with >75% radiolabeling yield. The radiocomplex was purified through the tc18 cartridge to obtain 99.89% radiochemical yield. IC-50 of the NAC-ATRIDAT conjugate was 0.8 mM in A549 cells as evaluated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazonium bromide assay. Binding affinity experiments on A549 cells showed noteworthy binding with KD in the nanomolar range. A time course study showed a Km value of 0.19 µM and Vmax value of 0.49 pmol/µg protein/min showing reasonable tumor kinetics. Efflux studies showed that the synthesized radioligand is transported majorly by LAT followed by the ASC system. Clearance was found to be renal with 7.67 ± 1.48% ID/g uptake at 30 min which substantially declined to 0.52 ± 0.% ID/g at 4 h. A significant uptake of 10.06 ± 1.056% ID/g was observed at the tumor site in mice at 1 h. µPET images revealed a high contrast with a tumor-to-kidney ratio of 4.8 and a tumor-to-liver ratio of 35.85 at 1 h after injection. These preclinical in vitro and in vivo evaluation supports its potential on the way of becoming a successful 68Ga-radiolabeled amino acid-based PET imaging agent.

Rose Bengal attached and dextran coated gadolinium oxide nanoparticles for potential diagnostic imaging applications.

We report here, reverse micelle mediated synthesis of multifunctional dextran (dex) coated Gd2O3 nanoparticles (NPs) carrying rose bengal (RB) dye for magnetic resonance and optical imaging. The diameter of these RB attached dex coated Gd2O3 NPs (Gd-dex-RB NPs) was found to be ~17 nm as measured by TEM. NMR line broadening effect on the surrounding water protons affirmed the paramagnetic nature of these NPs. Optical properties of Gd-dex-RB NPs were validated by UV-Vis and fluorescence spectroscopy. Time dependent release profile of RB from NPs at two different pH of 7.4 and 5.0 revealed that these NPs behave as slow releasing system. In-vitro study revealed that NPs are efficiently taken up by cells and show optical activity in cellular environment. In vitro cell viability (SRB) assay was performed on cancerous (A-549, U-87) and normal (HEK-293) cell lines, showed the absence of cytotoxic effect of Gd-dex-RB NPs. Therefore, such multifunctional NPs can be efficiently used for bio-imaging and optical tracking.

PEG coated and doxorubicin loaded multimodal Gadolinium oxide nanoparticles for simultaneous drug delivery and imaging applications.

We report water-in-oil microemulsion mediated synthesis of PEG1 coated Gd2O3 NPs2 loaded with fluorescent anti-cancer drug dox3 for synchronous drug delivery, optical and MR4 imaging applications. These PEG covered Gd2O3 NPs loaded with dox (Gd-PEG-dox NPs) were found to possess spherical morphology with 13nm size as measured from TEM and the hydrodynamic diameter comes out to be 37nm as determined from DLS. Fluorescence spectra and fluorescence microscopy images confirmed optical activity of the NPs. The paramagnetic nature of NPs was affirmed by NMR line broadening effect on the spectrum of surrounding water protons. Therefore, these particles can be efficiently used as CA5 in MR imaging. In vitro analysis showed significant cellular uptake of particles by A-549 cells. A pH dependent drug release pattern was observed for the NPs. Cell viability assay performed on A-549, PANC-1 and U-87 cancerous cell lines revealed that Gd-PEG-dox NPs are cytotoxic. On the basis of these observations, it can be concluded that these multi-modal paramagnetic NPs promise potential cancer therapy along with optical and MR imaging applications.

Biotinidase Resistant 68Gallium-Radioligand Based on Biotin/Avidin Interaction for Pretargeting: Synthesis and Preclinical Evaluation.

A new macrocyclic system 2,2'-(12-amino-11,13-dioxo-1,4,7,10-tetraazacyclotridecane-4,7-diyl)diacetic acid (ATRIDAT) was designed for coordinating metals in +2 and +3 oxidation states particularly 68Ga(III), for PET imaging. ATRIDAT was conjugated to d-biotin for pretargeting via biotin-avidin interaction. This model provides high tumor targeting efficiency and stability to biotinidase activity leading to modest signal amplification at the tumor site. Cyclization of triethylenetetramine with protected diethylamino malonate resulted in the formation of 13 membered diamide ring. d-Biotin was then anchored on the pendant amine rendering α-methyne carbon to the biotinamide bond which blocks the biotinidase enzyme activity. Biotinidase stability assay showed remarkable stability toward the action of biotinidase with ∼95% remaining intact after treatment following 4 h. Binding affinity experiments such as HABA assay, competitive displacement studies with d-biotin and CD showed high binding affinity of the molecule with avidin in nanomolar range. Biotin conjugate was successfully radiolabeled with 68Ga(III) with radiolabeling efficiency of ∼70% and then purified to get 99.9% radiochemical yield. IC50 of the compound was found to be 2.36 mM in HEK cell line and 0.82 mM in A549 as assessed in MTT assay. In biodistribution studies, the major route of excretion was found to be renal. Significant uptake of 4.15 ± 0.35% was observed in tumor in the avidin pretreated mouse at 1 h. µPET images also showed a high tumor to muscle ratio of 26.8 and tumor to kidney ratio of 1.74 at 1 h post-injection after avidin treatment.

FITC-Dextran entrapped and silica coated gadolinium oxide nanoparticles for synchronous optical and magnetic resonance imaging applications.

We report, microemulsion mediated synthesis of FITC-dextran dye entrapped and silica coated Gd2O3 nanoparticles (NPs) for dual purpose of optical and magnetic resonance imaging, in the present study. TEM image revealed that the average size of the NPs is 18nm and hydrodynamic diameter of the particles as measured by DLS comes out to be about 16nm. Gd2O3 core show paramagnetism which is affirmed by the NMR line broadening effect on neighboring water proton spectrum and also by magnetization curve obtained in VSM analysis. The fluorescence of the entrapped dye is confirmed by the UV-vis and fluorescence spectroscopy. Nanoencapsulation of FITC-dextran fluorophore was found to increase its optical activity and provided a blanket against quenching. Moreover, TGA data revealed that entrapment of dye imparts thermal stability to it and enhances its fluorescence in comparison to bare dye. The release kinetic pattern (at pH 7.4) of the entrapped dye revealed that these particles behave as non-releasing system. The in-vitro cell viability (SRB) assay of the particles done on normal cell line (HEK-293) as well as cancerous cell line (A-549) indicated non-cytotoxic nature of the particles. In a nut-shell, these particles have the potential to be efficiently used for optical and magnetic resonance imaging. We anticipate that further optimization of these particles can be done by either conjugating or entrapping a drug for targeted drug delivery which would open more prospective options in biomedical field.

Preclinical Evaluation of a Potential GSH Ester Based PET/SPECT Imaging Probe DT(GSHMe)2 to Detect Gamma Glutamyl Transferase Over Expressing Tumors.

Gamma Glutamyl Transferase (GGT) is an important biomarker in malignant cancers. The redox processes ensuing from GGT-mediated metabolism of extracellular GSH are implicated in critical aspects of tumor cell biology. Reportedly, Glutathione monoethyl ester (GSHMe) is a substrate of GGT, which has been used for its rapid transport over glutathione. Exploring GGT to be an important target, a homobivalent peptide system, DT(GSHMe)2 was designed to target GGT-over expressing tumors for diagnostic purposes. DT(GSHMe)2 was synthesized, characterized and preclinically evaluated in vitro using toxicity, cell binding assays and time dependent experiments. Stable and defined radiochemistry with 99mTc and 68Ga was optimized for high radiochemical yield. In vivo biodistribution studies were conducted for different time points along with scintigraphic studies of radiolabeled DT(GSHMe)2 on xenografted tumor models. For further validation, in silico docking studies were performed on GGT (hGGT1, P19440). Preclinical in vitro evaluations on cell lines suggested minimal toxicity of DT(GSHMe)2 at 100 µM concentration. Kinetic analysis revealed transport of 99mTc-DT(GSHMe)2 occurs via a saturable high-affinity carrier with Michaelis constant (Km) of 2.25 µM and maximal transport rate velocity (Vmax) of 0.478 µM/min. Quantitative estimation of GGT expression from western blot experiments showed substantial expression with 41.6 ± 7.07 % IDV for tumor. Small animal micro PET (Positron Emission Tomography)/CT(Computed Tomography) coregistered images depicted significantly high uptake of DT(GSHMe)2 at the BMG-1 tumor site. ROI analysis showed high tumor to contra lateral muscle ratio of 9.33 in PET imaging studies. Avid accumulation of radiotracer was observed at tumor versus inflammation site at 2 h post i.v. injection in an Ehrlich Ascites tumor (EAT) mice model, showing evident specificity for tumor. We propose DT(GSHMe)2 to be an excellent candidate for prognostication and tumor imaging using PET/SPECT.