Phase 3 RCT comparing docetaxel-platinum with docetaxel-platinum-5FU as neoadjuvant chemotherapy in borderline resectable oral cancer.

BACKGROUND: Neoadjuvant chemotherapy (NACT) with TPF (docetaxel, cisplatin, and 5FU) is one of the treatment options in very locally advanced oral cancer with a survival advantage over PF (cisplatin and 5FU). TP (docetaxel and cisplatin) has shown promising results with a lower rate of adverse events but has never been compared to TPF. METHODS: In this phase 3 randomized superiority study, adult patients with borderline resectable locally advanced oral cancers were randomized in a 1:1 fashion to either TP or TPF. After the administration of 2 cycles, patients were evaluated in a multidisciplinary clinic and further treatment was planned. The primary endpoint was overall survival (OS) and secondary endpoints were progression-free survival (PFS) and adverse events. RESULTS: 495 patients were randomized in this study, 248 patients in TP arm and 247 in TPF arm. The 5-year OS was 18.5% (95% CI 13.8-23.7) and 23.9% (95% CI 18.1-30.1) in TP and TPF arms, respectively (Hazard ratio 0.778; 95% CI 0.637-0.952; P = 0.015). Following NACT, 43.8% were deemed resectable, but 34.5% underwent surgery. The 5-year OS was 50.7% (95% CI 41.5-59.1) and 5% (95%CI 2.9-8.1), respectively, in the surgically resected versus unresected cohort post NACT (P < 0.0001). Grade 3 or above adverse events were seen in 97 (39.1%) and 179 (72.5%) patients in the TP and TPF arms, respectively (P < 0.0001). CONCLUSION: NACT with TPF has a survival benefit over TP in borderline resectable oral cancers, with an increase in toxicity which is manageable. Patients who undergo surgery achieve a relatively good, sustained survival.

A Comparative Study Evaluating the Quality of Life and Survival Outcomes in Patients Receiving Chemotherapy Versus Oral Tyrosine Kinase Inhibitor in the Third Line and Beyond Setting for Advanced NSCLC.

Introduction: The outcomes in advanced NSCLC have improved owing to the availability of more effective systemic and improved supportive care. This has increased the number of patients who seek treatment in the third line and beyond setting. We conducted this study to compare the quality of life (QoL), toxicity, and outcomes in patients receiving chemotherapy and EGFR tyrosine kinase inhibitors (TKIs) in this setting. Methods: In this phase 3, randomized, open-label study, patients with stage III or IV NSCLC with disease progression on at least two prior lines of chemotherapy, with a life expectancy of at least 3 months, without prior EGFR TKI exposure, and stable brain metastases (if any) were included. Patients were randomized to receive chemotherapy (gemcitabine or docetaxel or paclitaxel or vinorelbine) or an EGFR TKI (erlotinib or gefitinib). The primary end point was the change in QoL at 8 to 10 weeks; the secondary outcomes were safety and overall survival (OS). Patients underwent clinical evaluation at every visit, and toxicity was assessed as per Common Terminology Criteria for Adverse Events version 4.03. A radiological tumor response assessment was done every 8 to 12 weeks from the start of therapy. The QoL was assessed using the EORTC QLQ C30 and LC13 questionnaires. The change in QoL scores was calculated as the difference between scores at baseline and scores at 8 to 10 weeks (Δ) for each QoL domain. The Mann-Whitney U test was used to compare the mean difference (Δ) for each domain. OS and progression-free survival (PFS) were determined using the Kaplan-Meier method and Cox proportional regression analysis. Results: A total of 246 patients were enrolled in the study, with 123 in each arm. There was a male predominance with 69.1% male patients in the chemotherapy arm and 70.7% in the EGFR TKI arm. The median age of patients in the chemotherapy arm was 54 years and 55 years in the chemotherapy and EGFR TKI arms, respectively. There was no significant difference in the change in QoL at baseline and the second visit (Δ) in both arms in all domains of EORTC QLQ C30 except cognitive function (p = 0.0045) and LC13 except alopecia (0.01249). The mean Δ Global Health Status was -28 in the chemotherapy arm and -26.8 in the EGFR TKI arm; this was not statistically significant (p = 0.973). The median follow-up was 88.1 months (95% confidence interval [CI]: 39.04-137.15). On the intention-to-treat analysis, the median PFS was 3.13 months (95% CI: 2.15-4.11) in the chemotherapy arm and 2.26 months (95% CI: 2.1-2.43) in the EGFR TKI arm, with hazard ratio at 1.074 (95% CI: 0.83-1.38) (p = 0.58). There were 120 deaths in each arm. The median OS was 7.63 months (95% CI: 5.96-9.30) in the chemotherapy arm and 7.5 months in the EGFR TKI arm (95% CI: 5.85-9.14); hazard ratio at 1.033 (95% CI: 0.80-1.33) (p = 0.805). The toxicity profile was similar in both arms except for a significantly higher incidence of fatigue (p = 0.043), peripheral neuropathy (0.000), alopecia, hypokalemia (0.037), and pedal edema (0.007) in the chemotherapy arm and dry skin (p = 0.000) and skin rash (p = 0.019) in the EGFR TKI arm. Conclusions: There was no significant difference in most QoL scales (except cognitive function and alopecia), OS, and PFS of patients with advanced NSCLC receiving an EGFR TKI as compared with chemotherapy TKI in the third-line setting. The toxicity profile is consistent with the known toxicities of the agents.

Recurrent idiopathic pancreatitis complicating as emphysematous pancreatitis and gastroduodenal artery pseudoaneurysm: A rare case report.

Emphysematous pancreatitis (EP) is a rare and potentially fatal condition of the pancreas. It is associated with gas-forming bacteria and is characterized by the presence of gas in or around the pancreas. It is identified by a computed tomography scan of the abdomen. Although predisposing factors are not precisely known, diabetes mellitus, which predisposes to gas gangrene, is seen to be commonly associated with patients of EP. EP being potentially fatal requires immediate management. Surgery is generally indicated in EP. However, EP can also managed conservatively. In our case, the patient developed recurrent pancreatitis, the cause being idiopathic, and the second episode of acute pancreatitis was complicated by EP and gastroduodenal artery pseudoaneurysm.

Résumé La pancréatite emphysémateuse (EP) est une condition rare et potentiellement mortelle du pancréas. Il est associé à des bactéries de formation de gaz et se caractérise par la présence de gaz dans ou autour du pancréas. Il est identifié par une tomodensitométrie calculée de l'abdomen. Bien que les facteurs prédisposants ne soient pas précisément connus, le diabète sucré, qui prédispose à la gangrène du gaz, est considéré comme communément associé aux patients du PE. EP étant potentiellement mortel nécessite une gestion immédiate. La chirurgie est généralement indiquée dans EP. Cependant, EP peut également gérer de manière conservatrice. Dans notre cas, le patient a développé une pancréatite récurrente, la cause étant idiopathique, et le deuxième épisode de pancréatite aiguë a été compliqué par l'EP et le pseudo-anévrisme de l'artère gastroduodénale. Mots-clés: Pancréatite emphysémateuse, pseudo-anévrisme gastroduodénal, pancréatite.

Modified FOLFIRINOX compared to Gemcitabine & nab-Paclitaxel in advanced pancreatic ductal adenocarcinoma - results of a match-pair analysis.

Background & objectives: FOLFIRINOX and gemcitabine plus nab-paclitaxel (GN) are the most commonly used regimens in advanced pancreatic ductal adenocarcinomas (PDACs). As there is limited data on comparison of these two regimens, the present study was aimed to compare survivals and tolerance for both regimens through a match-pair analysis. Methods: The data of 350 patients with metastatic and locally advanced PDAC, treated between January 2013 and December 2019, were retrieved. A 1:1 matching, using age and performance status, without replacement was performed by using nearest neighbour matching method. Results: A total of 260 patients (130 modified FOLFIRINOX and 130 GN) were matched. The median overall survival (OS) was 12.98 months [95% confidence interval (CI) 7.257-8.776 months] in modifications of FOLFIRINOX (mFOLFIRINOX) cohort and 12.06 months (95% CI 6.690-8.88 months) in GN group (P=0.080). The incidence of grade 3 and 4 infections, diarrhoea, oral mucositis, and fatigue was higher with mFOLFIRINOX. Patients who received second line therapy had improved OS as compared to those who did not (14.06 vs. 9.07 months, P<0.001). Interpretation & conclusions: GN and mFOLFIRINOX appear to have similar survival outcomes in an unselected match paired patient population with advanced PDAC. A markedly increased incidence of non-myelosuppressive grade 3 and grade 4 side-effects and lack of survival improvements suggest a need for nuanced use of the mFOLFIRINOX regimen. Administration of second-line chemotherapy improves OS in patients with advanced PDAC.

Proteogenomic identification of Hepatitis B virus (HBV) genotype-specific HLA-I restricted peptides from HBV-positive patient liver tissues.

The presentation of virus-derived peptides by HLA class I molecules on the surface of an infected cell and the recognition of these HLA-peptide complexes by, and subsequent activation of, CD8+ cytotoxic T cells provides an important mechanism for immune protection against viruses. Recent advances in proteogenomics have allowed researchers to discover a growing number of unique HLA-restricted viral peptides, resulting in a rapidly expanding repertoire of targets for immunotherapeutics (i.e. bispecific antibodies, engineered T-cell receptors (TCRs), chimeric antigen receptor T-cells (CAR-Ts)) to infected tissues. However, genomic variability between viral strains, such as Hepatitis-B virus (HBV), in combination with differences in patient HLA alleles, make it difficult to develop therapeutics against these targets. To address this challenge, we developed a novel proteogenomics approach for generating patient-specific databases that enable the identification of viral peptides based on the viral transcriptomes sequenced from individual patient liver samples. We also utilized DNA sequencing of patient samples to identify HLA genotypes and assist in target selection. Liver samples from 48 HBV infected patients, primarily from Asia, were examined to reconstruct patient-specific HBV genomes, identify regions within the human chromosomes targeted by HBV integrations and obtain a comprehensive view of HBV peptide epitopes using our HLA class-I (HLA-I) immunopeptidomics discovery platform. Two previously reported HLA associated HBV-derived peptides, HLA-A02 binder FLLTRILTI (S194-202) from the large surface antigen and HLA-A11 binder STLPETTVVRR (C141-151) from the capsid protein were validated by our discovery platform, but both were detected at very low frequencies. In addition, we identified and validated, using heavy peptide analogues, novel strain-specific HBV-HLA associated peptides, such as GSLPQEHIVQK (P606-616) and variants. Overall, our novel approach can guide the development of bispecific antibody, TCR-T, or CAR-T based therapeutics for the treatment of HBV-related HCC and inform vaccine development.

Treatment pattern and outcomes in de novo T790M-mutated non-small cell lung cancer.

Introduction: Limited data exists for non-small cell lung cancer (NSCLC) patients harbouring de novo T790M mutation. Methods: NSCLC patients, with de novo T790M, who registered at our institute between 01/03/2015 and 31/12/2019, were considered for retrospective analysis of treatment pattern and clinical outcomes, i.e., progression-free survival (PFS) and overall survival (OS). Results: Of 1,542 epidermal growth factor receptor (EGFR)-mutated patients, 40 (2.59%) had de novo T790M. Most were male (27, 67.5%) and smokers (23, 57.5%). The commonest site of metastasis was the lungs (31, 77.5%), while 7 (17.5%) had central nervous system (CNS) involvement. Additional EGFR gene mutations and anaplastic lymphoma kinase (ALK) positivity were observed in 20 (50.0%) and 4 (10.0%) cases, respectively. The first-line systemic therapy and the number of patients receiving it were as follows: osimertinib by 14 (35.0%), first-generation EGFR tyrosine kinase inhibitors (TKIs) by 10 (25.0%), gefitinib + chemotherapy by 3 (7.5%), chemotherapy by 7 (17.5%) and gefitinib + bevacizumab by 2 (5%). One patient defaulted before starting any treatment. Hence, 39 were considered for survival analysis. The median PFS and OS for the entire cohort were 10.4 (95% CI = 7.6-19.7) months and 24.9 (95% CI = 15.7-NA) months, respectively. The median PFS for patients on osimertinib was 19.8 (95% CI = 11.6-28.0) months versus 8.8 (95% CI = 6.6-10.9) months for those on other systemic therapy. No CNS involvement, use of osimertinib or first-generation EGFR TKI plus chemotherapy or ALK inhibitor in ALK-positive cases prognosticated better PFS. When compared to other systemic therapies, osimertinib improved PFS in patients with or without additional EGFR mutations, although it was statistically significant for the former group only (p = 0.002). Conclusion: The incidence of de novo T790M is low. Osimertinib in frontline therapy provides promising outcomes.

High-Power Holmium with MOSES Technology or Thulium Fiber Laser in MiniPerc with Suction: A New Curiosity.

Purpose: To present initial clinical comparison between high-power holmium laser with MOSES technology (HPH-M) and thulium fiber laser (TFL) during mini-percutaneous nephrolithotomy (PCNL) for renal calculi with specific emphasis on fragmentation efficiency, fragment size distribution, and stone-free rates (SFRs). Materials and Methods: Between August 2018 and December 2019, we performed mini-PCNL for renal calculi <3 cm using HPH-M (Lumenis, Israel) or TFL (Urolase SP, IPG Photonics). Data were collected prospectively in our institutional stone registry. Propensity score matching (1:1) was performed using stone size and density as predictors resulting in matched cohort of 51 patients in each group. Mini-PCNL with active suction sheath was standard across all patients. Primary end-point was SFR at immediate postprocedure and 1 month using CT/kidney, ureter, and bladder radiograph. Stone fragments were retrieved and segregated to assess proportion of dust (<1 mm), small (1-3 mm), and large (>3 mm) fragments. Result: Both groups were comparable in terms of stone size (p = 0.74), volume (p = 0.17), and density (p = 0.69). SFR at 48 hours was 78.43% in HPH-M group and 68.63% in TFL group. Patients with residual fragments were completely clear at 1 month. Lasing time (678.6 vs 551.95 seconds; p = 0.17), stone fragmentation rate (4.6 vs 5.2 mm3/s; p = 0.23), and total laser energy (21.9 vs 16.3 KJ; p = 0.09) were comparable in both arms. Both groups produced similar dusting (46.8 vs 46.41%; p = 0.93). TFL produced a greater proportion of fragments >3 mm (36% vs 22.68%, p = 0.002). On subset analysis based on stone density, all outcome parameters were comparable except a shorter total operative time with TFL (p ≤ 0.05). Conclusion: HPH-M and TFL showed similar SFR. Within constraints of the laser fiber size and energy settings, both modalities were equivalent in terms of fragmentation efficiency and proportion of dusting across stone densities.

A prospective comparative study of mini-PCNL using Trilogy™ or thulium fibre laser with suction.

INTRODUCTION: There has been recent introduction of novel lithotripters and high-power lasers for stone disintegration. With miniaturization of PCNL, there is need of effective disintegration and faster stone-clearance. This study aimed to evaluate efficiency of Trilogy™ and Thulium fibre laser (TFL) in mini-percutaneous nephrolithotomy (mini-PCNL). METHODS: This is prospective study comparing efficiency and outcomes of Trilogy™ and TFL in mini-PCNL between January 2019 and February 2020. Primary objective was to compare stone fragmentation rates, with secondary objectives beings stone-free rates and complications. RESULTS: There were 60 mini-PCNL with suction using either Trilogy™ or TFL energy source. Mean stone size and density were 27.60 ± 10.17 mm, 22.04 ± 9.69 mm (p = 0.05) and 1172.9 ± 313.5HU, 1308.9 ± 333.9HU (p = 0.10) for Trilogy™ and TFL, respectively. Using 3D doctor imaging software from CT images, mean stone volumes were 3718.9 ± 3038.7mm3 for Trilogy™ and 3425.9 ± 3096.1mm3 for TFL(p = 0.77). Using probe-activation time or lasing time, stone-fragmentation rate was 5.98 ± 4.25mm3/sec for Trilogy™ and 3.95 ± 1.00mm3/sec for TFL(p = 0.015). Treatment time (puncture to complete clearance) was 32.48 ± 15.39 min for Trilogy™ and 28.63 ± 18.56 min for TFL(p = 0.38). Haemoglobin drop was 1.19 ± 0.76gm/dl for Trilogy™ and 0.99 ± 0.74gm/dl for TFL (p = 0.30). Trilogy™ arm had 96.6% complete clearance and TFL had 76.6% in TFL at 48 h. One patient in Trilogy™ arm required auxiliary RIRS for residual stone. Both arms had complete stone clearance at 1 month follow-up. Trilogy™ arm had 3 Clavien-Dindo grade-II complications while TFL had 2 Clavien-Dindo grade-II complications (UTI requiring antibiotics). There was no blood transfusion in either of arm. CONCLUSION: Trilogy™ had significantly better stone fragmentation rate than TFL in managing renal stones. However, stone-free rates and complications were comparable for Trilogy™ and TFL.

Size Distribution of Fragments by High-power Holmium Laser Lithotripsy in MiniPCNL with Suction.

PURPOSE OF REVIEW: Data are lacking on distribution of size of fragments created with the laser lithotripsy, stone density, and composition. Dust (< 1 mm) and small fragments (1-3 mm) may be too small to be efficiently removed with forceps compared to larger fragments (> 3 mm). We aim to report the size distribution of fragments formed during holmium laser lithotripsy. RECENT FINDINGS: One hundred ten patients with renal calculi < 3 cm underwent miniPCNL using ClearPetra sheath (Well Lead Medical Co., China) with controllable irrigation and suction system that minimizes fragment dispersion and maximizes fragment aspiration. Moses holmium laser (Lumenis INC, Israel) was used with predefined laser energy settings for lithotripsy. Proportion of fragment size < 1 mm was 46.36 ± 16.68%, 1-3 mm was 28.18 ± 10.01%, and > 3 mm was 25.19 ± 16.18% for the entire cohort. Complete stone clearance at 48 h was achieved in 77.3% cases. The remaining 22.7% patients had complete clearance at 1-month follow-up CT. In all the stone density, volume, and composition groups, majority of fragments created were either < 1 mm or 1-3 mm. Only 25% fragments were > 3 mm that would get aspirated out through the sheath with suction or could be retrieved with forceps. The combination of a high-power holmium laser and suction would help in complete stone clearance with effective aspiration of smaller stone fragments and dust simultaneously during lasing.

An observational study to see the correlation between trends of platelet counts and immature platelet fraction in dengue infection.

Dengue, a common tropical viral disease, often has complications of thrombocytopenia causing bleeding and warranting blood transfusion. This is costly in low-income settings. We conducted an observational study using a relatively new parameter called immature platelet fraction which indicates regeneration of platelets by the bone marrow. Our study on 124 dengue patients showed a strong correlation between platelet count and immature platelet fraction and we observed that 96.1% and 97.4% patients showed rise in platelet count at 24 and 48 h, respectively. In the absence of bleeding, platelet transfusion was prevented in 64% of patients with an IPF level of 10% or more.

A clinical experience of thulium fibre laser in miniperc to dust with suction: a new horizon.

PURPOSE: To report safety and efficacy of mini-PCNL with suction attached to sheath combined with high-power Thulium Fibre laser (TFL). The secondary aim was to evaluate optimal laser settings for maximum stone dusting. MATERIALS AND METHODS: Prospective, single arm study was conducted from June 2019-December 2019 using miniPCNL with suction and TFL in 54 patients with renal stones < 3 cm. Stone fragments for each laser setting were independently retrieved and segregated according to size(< 1 mm,1-3 mm, > 3 mm) and weighed. Xray/CT scan imaging was performed in all patients within 48 h and 30 days to assess stone clearance. Optimal laser settings were evaluated for maximum dusting. RESULTS: Mean stone size was 18.32 ± 6.37 mm, volume was 2337.75 ± 1996.84mm3 and stone density was 1300.55 ± 435.32 HU. Total operative time was 39.85 ± 20.52 min, laser time was 10.08 ± 7.41 min and stone fragmentation rate was 5.02 ± 3.93 mm3/s. The procedure was completely tubeless in 37.04%, nephrostomy tube in 37.04% and DJ stent placed in 25.92%. Postoperatively, three patients had urinary infection (Clavien 2). Complete stone clearance at 48 h was achieved in 35 (64.8%) cases. 19 patients (35.2%) who had residual fragments at 48 h, had 100% clearance at one month on CT/Xray KUB. CONCLUSIONS: MiniPCNL using a nephrostomy sheath with suction along with high power Thulium Fibre Laser is safe and effective modality for lithotripsy. An initial laser setting of 0.2 J and 125-200 Hz was optimal for maximum dusting and simultaneous aspiration. Randomized comparative studies with other energy sources are being considered.

Mandibular incisor extraction treatment in Angle's Class I malocclusion with peg-shaped maxillary lateral incisors.

Accurate diagnosis and treatment planning are essential for obtaining ideal treatment result in cases involving mandibular incisor extraction. This case report describes a 15-year-old female with balanced soft-tissue profile, peg-shaped maxillary lateral incisors, and moderate mandibular anterior crowding treated with a mandibular incisor extraction. Ideal overbite and overjet were achieved. "Black triangle" formation was avoided due to the bodily movement of mandibular incisors and the use of uprighting springs for ideal axial inclination of mandibular incisors. A mandibular incisor extraction can be an effective treatment option in carefully selected clinical situations.

Myostatin deficiency but not anti-myostatin blockade induces marked proteomic changes in mouse skeletal muscle.

Pharmacologic blockade of the myostatin (Mstn)/activin receptor pathway is being pursued as a potential therapy for several muscle wasting disorders. The functional benefits of blocking this pathway are under investigation, in particular given the findings that greater muscle hypertrophy results from Mstn deficiency arising from genetic ablation compared to post-developmental Mstn blockade. Using high-resolution MS coupled with SILAC mouse technology, we quantitated the relative proteomic changes in gastrocnemius muscle from Mstn knockout (Mstn(-/-) ) and mice treated for 2-weeks with REGN1033, an anti-Mstn antibody. Relative to wild-type animals, Mstn(-/-) mice had a two-fold greater muscle mass and a >1.5-fold change in expression of 12.0% of 1137 quantified muscle proteins. In contrast, mice treated with REGN1033 had minimal changes in muscle proteome (0.7% of 1510 proteins >1.5-fold change, similar to biological difference 0.5% of 1310) even though the treatment induced significant 20% muscle mass increase. Functional annotation of the altered proteins in Mstn(-/-) mice corroborates the mutiple physiological changes including slow-to-fast fiber type switch. Thus, the proteome-wide protein expression differs between Mstn(-/-) mice and mice subjected to specific Mstn blockade post-developmentally, providing molecular-level insights to inform mechanistic hypotheses to explain the observed functional differences.

Computational fluid dynamics for the assessment of upper airway changes in skeletal Class III patients treated with mandibular setback surgery.

OBJECTIVE: To analyze and compare pharyngeal airflow characteristics pre- and post-mandibular setback surgery in patients with Class III skeletal dysplasia using cone beam computed tomography (CBCT) and computational fluid dynamics (CFD). MATERIALS AND METHODS: Records of 29 patients who had received orthodontic treatment along with mandibular setback surgery were obtained. CBCT scans were obtained at three time points: T1 (before surgery), T2 (average of 6 months after surgery), and T3 (average of 1 year after surgery). Digitized pharyngeal airway models were generated from these scans. CFD was used to simulate and characterize pharyngeal airflow. RESULTS: Mean airway volume was significantly reduced from 35,490.324 mm3 at T1 to 24,387.369 mm3 at T2 and 25,069.459 mm3 at T3. Significant increase in mean negative pressure was noted from 3.110 Pa at T1 to 6.116 Pa at T2 and 6.295 Pa at T3. There was a statistically significant negative correlation between the change in airway volume and the change in pressure drop at both the T2 and T3 time points. There was a statistically significant negative correlation between the amount of mandibular setback and change in pressure drop at the T2 time point. CONCLUSIONS: Following mandibular setback surgery, pharyngeal airway volume was decreased and relative mean negative pressure was increased, implying an increased effort required from a patient for maintaining constant pharyngeal airflow. Thus, high-risk patients undergoing a large amount of mandibular setback surgery should be evaluated for obstructive sleep apnea and the proposed treatment plan be revised based on the risk for potential airway compromise.

Discovery of potential and selective COX-1 inhibitory leads using pharmacophore modelling, in silico screening and in vitro evaluation.

Cyclooxygenase -1 (COX-1) selective inhibitors are anticipated to be potential therapeutic agents for thrombosis, tumorigenesis, atherosclerosis, neuroprotection, and oxidative stress. In this study, a 3D-QSAR pharmacophore model was developed for potent and selective COX-1 inhibition based on 44 compounds from four different scaffolds using Phase, Schrödinger. One (hydrogen-bond) acceptor, one hydrophobic, and two aromatic sites (AHRR) contribute to COX-1 inhibitory activity. Test and decoy sets were used to corroborate the best hypothesis and the validated hypothesis was used to screen the SPECS database. The resultant hits were filtered by standard precision (SP) and extra precision (XP) modes of docking using Glide, Schrödinger which yielded five hits. Free energy calculations were carried out to quantify the affinity differences of the hits towards COX enzymes. These five hits were subjected to in vitro COX (ovine) inhibitory activity studies. The hits displayed potent COX-1 inhibitory activity and good selectivity versus COX-2 enzyme. The compounds also protected the nitric oxide (NO) induced cell death mediated by COX-1 in mouse macrophages cell line. Hence, we hypothesize that these compounds could be promising leads for the design of superior COX-1 inhibitors and insights gained from further exploration of the same could provide pertinent clues for the treatment of the conditions mentioned above.

Microwave and magnetic (M(2) ) proteomics of the experimental autoimmune encephalomyelitis animal model of multiple sclerosis.

We hypothesized that quantitative MS/MS-based proteomics at multiple time points, incorporating rapid microwave and magnetic (M(2) ) sample preparation, could enable relative protein expression to be correlated to disease progression in the experimental autoimmune encephalomyelitis (EAE) animal model of multiple sclerosis. To test our hypothesis, microwave-assisted reduction/alkylation/digestion of proteins from brain tissue lysates bound to C8 magnetic beads and microwave-assisted isobaric chemical labeling were performed of released peptides, in 90 s prior to unbiased proteomic analysis. Disease progression in EAE was assessed by scoring clinical EAE disease severity and confirmed by histopathologic evaluation for central nervous system inflammation. Decoding the expression of 283 top-ranked proteins (p <0.05) at each time point relative to their expression at the peak of disease, from a total of 1191 proteins observed in four technical replicates, revealed a strong statistical correlation to EAE disease score, particularly for the following four proteins that closely mirror disease progression: 14-3-3ε (p = 3.4E-6); GPI (p = 2.1E-5); PLP1 (p = 8.0E-4); PRX1 (p = 1.7E-4). These results were confirmed by Western blotting, signaling pathway analysis, and hierarchical clustering of EAE risk groups. While validation in a larger cohort is underway, we conclude that M(2) proteomics is a rapid method to quantify putative prognostic/predictive protein biomarkers and therapeutic targets of disease progression in the EAE animal model of multiple sclerosis.

Immunoenrichment microwave and magnetic proteomics for quantifying CD47 in the experimental autoimmune encephalomyelitis model of multiple sclerosis.

We hypothesized that quantitative MS/MS-based proteomics at multiple time points, incorporating immunoenrichment prior to rapid microwave and magnetic (IM(2) ) sample preparation, might enable correlation of the relative expression of CD47 and other low abundance proteins to disease progression in the experimental autoimmune encephalomyelitis (EAE) animal model of multiple sclerosis. To test our hypothesis, anti-CD47 antibodies were used to enrich for low abundance CD47 prior to microwave and magnetic proteomics in EAE. Decoding protein expression at each time point, with CD47-immunoenriched samples and targeted proteomic analysis, enabled peptides from the low abundance proteins to be precisely quantified throughout disease progression, including: CD47: 86-99, corresponding to the "marker of self" overexpressed by myelin that prevents phagocytosis, or "cellular devouring," by microglia and macrophages; myelin basic protein: 223-228, corresponding to myelin basic protein; and migration inhibitory factor: 79-87, corresponding to a proinflammatory cytokine that inhibits macrophage migration. While validation in a larger cohort is underway, we conclude that IM(2) proteomics is a rapid method to precisely quantify peptides from CD47 and other low abundance proteins throughout disease progression in EAE. This is likely due to improvements in selectivity and sensitivity, necessary to partially overcome masking of low abundance proteins by high abundance proteins and improve dynamic range.