Cancer cells display abnormal redox metabolism. Autophagy, anoikis and reactive oxygen species (ROS) play a regulatory role during metastasis. LC3 is a well-known essential molecule for autophagy. Therefore, we wanted to explore the molecular interplay between autophagy, anoikis, and ROS in relation to LC3B. We observed enhanced LC3B level along with increased expression of p62 and modulation of other autophagy-related molecules (Atg 3, 5, 7, 12, 16L1 and Beclin1) by inducing oxidative-stress in ovarian cancer cells using a ROS-producing pro-oxidant molecule. Surprisingly, enhanced LC3B was unable to induce autophagosome formation rather promoted anoikis. ROS-induced inhibition of autophagosome-formation is possibly due to the instability of autophagy initiator, ULK1 complex. Moreover, such upregulation of LC3B via ROS enhanced several apoptotic molecules. Silencing LC3B reduced these apoptotic molecules and increased when overexpressed, suggesting its role in apoptosis. Furthermore, LC3B-dependent apoptosis was decreased by inhibiting ROS, indicating a possible link between ROS, LC3B, and apoptosis. Additionally, ROS-induced enhanced LC3B promoted detachment-induced cell death (anoikis). This was further reflected by reduced cell adhesion molecules (integrin-β3 and focal adhesion kinase) and mesenchymal markers (snail and slug). Our in vitro experimental data was further confirmed in primary tumors developed in syngeneic mice, which also showed ROS-mediated LC3B enhancement along with reduced autophagosomes, integrin-β3 and focal adhesion kinase ultimately leading to the decreased tumor mass. Additionally, primary cells from high-grade serous carcinoma patient’s ascites exhibited LC3B enhancement and autophagy inhibition through ROS which provided a clinical relevance of our study. Taken together, this is the first evidence for a non-canonical role of LC3B in promoting anoikis in contrast to autophagy and may, therefore, consider as a potential therapeutic target molecule in ovarian cancer. Taken together, autophagy-inhibition may be an alternative approach to induce apoptosis/anoikis in cancer.

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5-Fluorouracil (5-FU) alone or in combination with other drugs is the main basis of chemotherapeutic treatment in colorectal cancer although patients with microsatellite instability generally show resistance to 5-FU treatment. The present investigation is focussed on the mechanistic insight of a pure herbal carbazole alkaloid, mahanine, as a single or in combination with 5-FU in colon cancer. We demonstrated that mahanine-induced apoptosis involved reactive oxygen species (ROS)-mediated nuclear accumulation of PTEN and its interaction with p53/p73. Mahanine and 5-FU in combination exerted synergistic inhibitory effect on cell viability. This combination also enhanced ROS production, increased tumour suppressor proteins and suppressed chemo-migration. Taken together, our results revealed that mahanine can be a potential chemotherapeutic agent with efficacy to reduce the concentration of toxic 5-FU in colon cancer.

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Black tea processing consists of four steps, namely withering, CTC, fermentation and drying, while cup quality made tea mostly controlled by fermentation step. This study evaluated biochemical changes at different stages of black tea processing in Bangladesh and determined the optimum fermentation time. Samples were collected from different tea processing stages to measure major phytochemicals and time intervals during fermentation to measure theaflavins and thearubigins ratio. Caffeine content was the least susceptible to processing steps. Biochemical changes started at withering, cell maceration and enzymatic oxidation started at CTC processing, thus the major reduction in the reducing sugar (20.46 to 04.95 ppm), catechin (16.88 to 7.95 ppm) and polyphenol (42.30 to 30.73 ppm) occurred here. The significant changes appeared during fermentation when polyphenol content decreased from 44.66 to 18.23 and catechin from 17.41 to 03.98 ppm due to the breakdown of these compounds to theaflavins (TF) and thearubigins (TR). The TF and TR ratio increased with fermentation time, and the highest of 1:8.4 was found at 50 min, which turned into 1:10 in the final product. The made tea quality parameters were comparable or better at fermentation time of 50 min than the quality of the black tea available in market.

T raumatic gastrointestinal tract (GIT) injuries are uncommon, representing less than 10% of all injuries in trauma patients (1), with a higher incidence in penetrating rather than blunt trauma (2). Small bowel is the most common site of alimentary tract injury in trauma followed by colorectal injuries, while gastroduodenal injuries comprise less than 2% of all injuries in trauma (3). In the current scenario of preferential nonoperative management in trauma, detection of GIT injury remains an important indication for urgent laparotomy as delayed diagnosis and surgery are associated with high morbidity and mortality (4). Also the type of operative management, prognosis, and outcome vary according to the number and site of GIT injuries (5).Multidetector computed tomography (MDCT) is a proven diagnostic modality for detection of traumatic GIT injuries with a reported sensitivity of 70%-95% and specificity of 92%-100% (6-11). Various CT signs of GIT and mesenteric injuries have been described such as bowel wall discontinuity, extraluminal air (either free or perivisceral), bowel wall thickening, abnormal bowel wall enhancement and intramural air, while mesenteric signs include mesenteric infiltration, active vascular contrast extravasation, beading, and abrupt termination of mesenteric vessels. These signs have reportedly variable incidences, sensitivities, and specificities; but if present, can lead prompt laparotomy and surgical exploration of the GIT to look for direct evidence of injury (12)(13)(14)(15)(16).MDCT with its capabilities of fast scanning, thin-section acquisition, and multiplanar reformatting has greatly enhanced the potential for detecting direct evidence of bowel in- 29From the Departments of Radiology (A.K.  dratinkumar@gmail.com; A.P., S.G., R.D., S.P.) and Surgery (A.G., S.K.), Jai Prakash Narayana Apex Trauma Centre, All India Institute of Medical Sciences, New Delhi, India. E M E R G E N C Y R A D I O LO G Y O R I G I N A L A R T I C L E PURPOSEWe aimed to assess the performance of computed tomography (CT) in localizing site of traumatic gastrointestinal tract (GIT) injury and determine the diagnostic value of CT signs in site localization. METHODSCT scans of 97 patients with surgically proven GIT or mesenteric injuries were retrospectively reviewed by radiologists blinded to surgical findings. Diagnosis of either GIT or mesenteric injuries was made. In patients with GIT injuries, site of injury and presence of CT signs such as focal bowel wall hyperenhancement, hypoenhancement, wall discontinuity, wall thickening, extramural air, intramural air, perivisceral infiltration, and active vascular contrast leak were evaluated. RESULTSOut of 97 patients, 90 had GIT injuries (70 single site injuries and 20 multiple site injuries) and seven had isolated mesenteric injury. The overall concordance between CT and operative findings for exact site localization was 67.8% (61/90), partial concordance rate was 11.1% (10/90), and discordance rate was 21.1% (19/90). For single site localization, concordance...

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Several drugs elicit their therapeutic efficacy by modulating multiple cellular targets and possess varied polypharmacological actions. The identification of the molecular targets of a potent bioactive molecule is essential in determining its overall polypharmacological profile. Experimental procedures are expensive and time-consuming. Therefore, computational approaches are actively implemented in rational drug discovery. Here, we demonstrate a computational pipeline, based on reverse virtual screening technique using several consensus scoring strategies, and perform structure-based kinase profiling of 12 FDA-approved drugs. This target prediction showed an overall good performance, with an average AU-ROC greater than 0.85 for most drugs, and identified the true targets even at the top 2% cutoff. In contrast, 10 non-kinase binder drugs exhibited lower binding efficiency and appeared in the bottom of ranking list. Subsequently, we validated this pipeline on a potent therapeutic molecule, mahanine, whose polypharmacological profile related to targeting kinases is unknown. Our target-prediction method identified different kinases. Furthermore, we have experimentally validated that mahanine is able to modulate multiple kinases that are involved in cross-talk with different signaling molecules, which thereby exhibits its polypharmacological action. More importantly, in vitro kinase assay exhibited the inhibitory effect of mahanine on two such predicted kinases' (mTOR and VEGFR2) activity, with IC values being ∼12 and ∼22 μM, respectively. Next, we generated a comprehensive drug-protein interaction fingerprint that explained the basis of their target selectivity. We observed that it is controlled by variations in kinase conformations followed by significant differences in crucial hydrogen-bond and van der Waals interactions. Such structure-based kinase profiling could provide useful information in revealing the unknown targets of therapeutic molecules from their polypharmacological behavior and would assist in drug discovery.