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Using a panel of cancer cell lines, we characterized a novel degrader of AKT, MS21. In mutant PI3K–PTEN pathway cell lines, AKT degradation was superior to AKT kinase inhibition for reducing cell growth and sustaining lower signaling over many days. AKT degradation, but not kinase inhibition, profoundly lowered Aurora kinase B (AURKB) protein, which is known to be essential for cell division, and induced G2–M arrest and hyperploidy. PI3K activated AKT phosphorylation of AURKB on threonine 73, which protected it from proteasome degradation. A mutant of AURKB (T73E) that mimics phosphorylation and blocks degradation rescued cells from growth inhibition. Degrader-resistant lines were associated with low AKT phosphorylation, wild-type PI3K/PTEN status, and mutation of KRAS/BRAF. Pan-cancer analysis identified that 19% of cases have PI3K–PTEN pathway mutation without RAS pathway mutation, suggesting that these patients with cancer could benefit from AKT degrader therapy that leads to loss of AURKB.Significance:MS21 depletes cells of phosphorylated AKT (pAKT) and a newly identified AKT substrate, AURKB, to inhibit tumor growth in mice. MS21 is superior to prior agents that target PI3K and AKT due to its ability to selectively target active, pAKT and sustain repression of signaling to deplete AURKB.This article is highlighted in the In This Issue feature, p. 2945

Supplementary Data from Loss of PBRM1 Alters Promoter Histone Modifications and Activates ALDH1A1 to Drive Renal Cell Carcinoma

Supplementary Figure from AKT Degradation Selectively Inhibits the Growth of PI3K/PTEN Pathway–Mutant Cancers with Wild-Type KRAS and BRAF by Destabilizing Aurora Kinase B

Supplementary Table from AKT Degradation Selectively Inhibits the Growth of PI3K/PTEN Pathway–Mutant Cancers with Wild-Type KRAS and BRAF by Destabilizing Aurora Kinase B

Supplementary Figure from Loss of PBRM1 Alters Promoter Histone Modifications and Activates ALDH1A1 to Drive Renal Cell Carcinoma

Supplementary Data from AKT Degradation Selectively Inhibits the Growth of PI3K/PTEN Pathway–Mutant Cancers with Wild-Type KRAS and BRAF by Destabilizing Aurora Kinase B

Figure S1: Expression of GSH synthesis enzymes in ccRCC tumors and adjacent healthy kidney tissue, demonstrating increased GGT1 levels in ccRCC. Figure S2: GGT1 mRNA and protein abundance in ccRCC and renal epithelial cell lines. Figure S3: GGT1 inhibition results in proliferation defect in ccRCC cells. Figure S4: GGT1 knockdown results in cell cycle arrest in ccRCC cell lines. Figure S5: GSH levels and GSH/GSSG ratios are reduced in GGT1 KD tumor xenografts, relative to controls. Figure S6: GSH pathway inhibition leads to proliferation defects in ccRCC cells.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer. While the localized form of this disease can be treated surgically, advanced and metastatic stages are resistant to chemotherapies. Although more innovative treatments, such as targeted or immune-based therapies, exist, the need for new therapeutic options remains. ccRCC presents unique metabolic signatures and multiple studies have reported a significant increase in levels of reduced glutathione (GSH) and its precursors in ccRCC tumor samples compared with normal kidney tissues. These observations led us to investigate the effects of blocking the GSH pathway, particularly the gamma-glutamyltransferase 1 (GGT1) enzyme, in multiple ccRCC cell lines. In this study, we provide in vitro and in vivo evidence that GGT1/GSH pathway inhibition impacts ccRCC cell growth, through increased cell-cycle arrest. Of note, GGT1 inhibition also impairs ccRCC cell migration. Finally, pharmacologic GSH pathway inhibition decreases ccRCC cell proliferation and increases sensitivity to standard chemotherapy. Our results suggest that GGT1/GSH pathway inhibition represents a new strategy to overcome ccRCC chemoresistance.Implications:GGT1/GSH pathway inhibition represents a promising therapeutic strategy to overcome chemoresistance and inhibit progression of ccRCC tumors.

The Covid pandemic has become a serious public health challenge for people across India and other nations. Nowadays, people rely on the online reviews being shared on different review sites to gather information about hospitals like the availability of beds, availability of ventilators, etc. However, since these reviews are large in number and are unstructured, patients struggle to get accurate and reliable information about the hospitals, due to which they end up taking admission into a hospital which might not be appropriate for the specific treatment they require. This paper employs the use of sentiment analysis to understand various online reviews of hospitals and provide valuable information to the patients. Approximately 30,000 + reviews were collected from more than 500 hospitals. The broad objective of the study is to give the patients a comprehensive and descriptive rating of the hospitals based on the online reviews given by different patients. In addition to providing a comprehensive summary, the study has conducted aspect-based analysis where it compares the hospitals based on four different aspects of the hospital viz. “Doctors’ services”, “Staff’s services”, “Hospital facilities”, and “Affordability”. The database containing aspect-based ratings of the hospitals will be of great value to the patients by allowing them to compare and select the best hospital based on the optimum fit of the aspects of their preference. [ FROM AUTHOR] Copyright of New Generation Computing is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full . (Copyright applies to all s.)

The need of the customers to be connected to the network at all times has led to the evolution of mobile technology. Operating systems play a vitol role when we talk of technology. Nowadays, Android is one of the popularly used operating system in mobile phones. Authors have analysed three stable versions of Android, 6.0, 7.0 and 8.0. Incorporating a change in the version after it is released requires a lot of rework and thus huge amount of costs are incurred. In this paper, the aim is to reduce this rework by identifying certain parts of a version during early phase of development which need careful attention. Machine learning prediction models are developed to identify the parts which are more prone to changes. The accuracy of such models should be high as the developers heavily rely on them. The high dimensionality of the dataset may hamper the accuracy of the models. Thus, the authors explore four dimensionality reduction techniques, which are unexplored in the field of network and communication. The results concluded that the accuracy improves after reducing the features.

Using a panel of cancer cell lines, we characterized a novel degrader of AKT, MS21. In mutant PI3K–PTEN pathway cell lines, AKT degradation was superior to AKT kinase inhibition for reducing cell growth and sustaining lower signaling over many days. AKT degradation, but not kinase inhibition, profoundly lowered Aurora kinase B (AURKB) protein, which is known to be essential for cell division, and induced G2–M arrest and hyperploidy. PI3K activated AKT phosphorylation of AURKB on threonine 73, which protected it from proteasome degradation. A mutant of AURKB (T73E) that mimics phosphorylation and blocks degradation rescued cells from growth inhibition. Degrader-resistant lines were associated with low AKT phosphorylation, wild-type PI3K/PTEN status, and mutation of KRAS/BRAF. Pan-cancer analysis identified that 19% of cases have PI3K–PTEN pathway mutation without RAS pathway mutation, suggesting that these patients with cancer could benefit from AKT degrader therapy that leads to loss of AURKB. Significance: MS21 depletes cells of phosphorylated AKT (pAKT) and a newly identified AKT substrate, AURKB, to inhibit tumor growth in mice. MS21 is superior to prior agents that target PI3K and AKT due to its ability to selectively target active, pAKT and sustain repression of signaling to deplete AURKB. This article is highlighted in the In This Issue feature, p. 2945

The study was conducted to evaluate the clinical outcomes of various glaucoma surgeries (trabeculectomy, trabeculectomy with mitomycin-C and glaucoma valve implantation) in cases of refractory glaucoma. When it becomes obvious that medication will not help then alternatives should be tried.These alternatives can be in form of :- Filtration Surgery Termed as trabeculectomy allows drainage of aqueous humor from within the eye to underneath the conjunctiva where it is absorbed. Trabeculectomy with mitomycin C-The use of antifibrotic agents such as mitomycin-C and 5-fluorouracil, along with releasable sutures or laser suture lysis, enhances the longevity of guarded procedures, Drainage Implants Glaucoma drainage devices are designed to divert aqueous humor from the anterior chamber to an external reservoir, where a fibrous capsule forms about 4-6 weeks after surgery and regulates flow. Aims of Study 1. Study the clinical outcomes of trabeculectomy,trabeculetomy with mitomycin-C and glaucoma valve implantation in management of refractory glaucoma patients. 2.Evaluate the complications of glaucoma valve surgery. 3.Study the complications of trabeculectomy,trabeculectomy with mitomycin-C. The proposed study was conducted in upgraded department of ophthalmology,LLRM Medical College,Meerut. Population characteristics were number of eyes treated,diagnosed and follow up.The study was a prospective study with random selection of patients done from the OPD.The randomly selected patients were then divided into three groups, one group was subjected to trabeculectomy, second with trabeculectomy with mitomycin-C and the other group was subjected to glaucoma valve implantation. CRITERIA FOR FAILURE OF SURGERY: Ÿ IOP > 21 mm hg at end of 1 year or Ÿ IOP not reduced by 20% from baseline at end of 1 year or Ÿ IOP

Tumors exhibit widespread enhancer landscape reprogramming compared to normal tissue. The etiology is believed to be largely cell-intrinsic in non-hormonal cancers, attributed to such genomic alterations as focal amplification of non-coding regions, aberrant activation of transcription factors, and non-coding mutations creating de novo transcription factor binding sites. Here, using freshly resected primary CRC tumors and patient-matched adjacent normal colon epithelia, we find divergent epigenetic landscapes between primary CRC tumors and CRC cell lines. We identify a unique super-enhancer signature largely absent in cell culture. Intriguingly, this phenomenon extends to highly recurrent aberrant super-enhancers gained in CRC over patient-matched normal epithelium suggesting novel insight into the etiology of enhancer reprogramming in CRC and its downstream relevance to tumor biology. We find one such super-enhancer activated in epithelial cancer cells due to surrounding inflammation in the tumor microenvironment. CRISPR-dcas9-KRAB interference of this super-enhancer identifies PDZK1IP1 as its target gene. PDZK1IP1 is previously observed to be highly up-regulated in CRC. However, the mechanism behind its transcriptional activation is not fully understood. We restore both the super-enhancer and PDZK1IP1 levels following treatment with cytokines or xenotransplantation into nude mice, thus demonstrating its etiology via local tumor microenvironment acquisition. Deletion of inflammatory transcription factors RELA and STAT3 in human CRC cells inhibits PDZK1IP1 induction in xenografts. PDZK1IP1 appears to be critical for CRC growth in the setting of its super-enhancer induction as xenografts, but not in cell culture where the super-enhancer is absent and expression is largely silent. Building on its known role in glucose uptake via SGLT receptors, we demonstrate mechanistically that PDZK1IP1 enhances the reductive capacity CRC cancer cells via the pentose phosphate pathway using polar metabolomic profiling. We show this activation enables efficient growth under oxidative conditions both in vitro and in vivo, challenging the previous notion that PDZK1IP1 acts as a tumor suppressor in CRC. Collectively, these observations highlight the biologic significance of epigenomic profiling on patient-matched primary specimens and identify this microenvironment-acquired super-enhancer as an oncogenic driver in the setting of the inflamed tumor. Citation Format: Royce Zhou, Jia Xu, Tiphaine Martin, Alexis Zachem, John He, Sait Ozturk, Deniz Demircioglu, Ankita Bansal, Andrew Trotta, Bruno Giotti, Berkley Gryder, Yao Shen, Saul Carcamo, Xuewei Wu, Kaitlyn Bosch, Benjamin Hopkins, Alexander Tsankov, Randolph Steinhagen, Drew Jones, John Asara, Jerry Chipuk, Rachel Brody, Steven Itzkowitz, Iok In Christine Chio, Dan Hasson, Emily Bernstein, Ramon Parsons. A local tumor microenvironment acquired super-enhancer induces an oncogenic driver in colorectal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3481.

Subunits of SWI/SNF chromatin remodeling complexes are frequently mutated in human malignancies. The PBAF complex is composed of multiple subunits, including the tumor-suppressor protein PBRM1 (BAF180), as well as ARID2 (BAF200), that are unique to this SWI/SNF complex. PBRM1 is mutated in various cancers, with a high mutation frequency in clear cell renal cell carcinoma (ccRCC). Here, we integrate RNA-seq, histone modification ChIP-seq, and ATAC-seq data to show that loss of PBRM1 results in de novo gains in H3K4me3 peaks throughout the epigenome, including activation of a retinoic acid biosynthesis and signaling gene signature. We show that one such target gene, ALDH1A1, which regulates a key step in retinoic acid biosynthesis, is consistently upregulated with PBRM1 loss in ccRCC cell lines and primary tumors, as well as non-malignant cells. We further find that ALDH1A1 increases the tumorigenic potential of ccRCC cells. Using biochemical methods, we show that ARID2 remains bound to other PBAF subunits after loss of PBRM1 and is essential for increased ALDH1A1 after loss of PBRM1, whereas other core SWI/SNF components are dispensable, including the ATPase subunit BRG1. In total, this study uses global epigenomic approaches to uncover novel mechanisms of PBRM1 tumor suppression in ccRCC. Implications: This study implicates the SWI/SNF subunit and tumor-suppressor PBRM1 in the regulation of promoter histone modifications and retinoic acid biosynthesis and signaling pathways in ccRCC and functionally validates one such target gene, the aldehyde dehydrogenase ALDH1A1.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer. While the localized form of this disease can be treated surgically, advanced and metastatic stages are resistant to chemotherapies. Although more innovative treatments, such as targeted or immune-based therapies, exist, the need for new therapeutic options remains. ccRCC presents unique metabolic signatures and multiple studies have reported a significant increase in levels of reduced glutathione (GSH) and its precursors in ccRCC tumor samples compared with normal kidney tissues. These observations led us to investigate the effects of blocking the GSH pathway, particularly the gamma-glutamyltransferase 1 (GGT1) enzyme, in multiple ccRCC cell lines. In this study, we provide in vitro and in vivo evidence that GGT1/GSH pathway inhibition impacts ccRCC cell growth, through increased cell-cycle arrest. Of note, GGT1 inhibition also impairs ccRCC cell migration. Finally, pharmacologic GSH pathway inhibition decreases ccRCC cell proliferation and increases sensitivity to standard chemotherapy. Our results suggest that GGT1/GSH pathway inhibition represents a new strategy to overcome ccRCC chemoresistance. Implications: GGT1/GSH pathway inhibition represents a promising therapeutic strategy to overcome chemoresistance and inhibit progression of ccRCC tumors.

Tumors exhibit widespread enhancer landscape reprogramming compared to normal tissue. The etiology is believed to be largely cell-intrinsic in non-hormonal cancers, attributed to such genomic alterations as focal amplification of non-coding regions, aberrant activation of transcription factors, and non-coding mutations creating de novo transcription factor binding sites. Here, using freshly resected primary CRC tumors and patient-matched adjacent normal colon epithelia, we find divergent epigenetic landscapes between primary CRC tumors and CRC cell lines. We identify a unique super-enhancer signature largely absent in cell culture. Intriguingly, this phenomenon extends to highly recurrent aberrant super-enhancers gained in CRC over patient-matched normal epithelium suggesting novel insight into the etiology of enhancer reprogramming in CRC and its downstream relevance to tumor biology. We find one such super-enhancer activated in epithelial cancer cells due to surrounding inflammation in the tumor microenvironment. We restore this super-enhancer and its expressed gene, PDZK1IP1, following treatment with cytokines or xenotransplantation into nude mice, thus demonstrating its etiology via local tumor microenvironment acquisition. Building on its known role in glucose uptake via SGLT receptors, we demonstrate mechanistically that PDZK1IP1 enhances the reductive capacity CRC cancer cells via the pentose phosphate pathway using polar metabolomic profiling. We show this activation enables efficient growth under oxidative conditions both in vitro and in vivo, challenging the previous notion that PDZK1IP1 acts as a tumor suppressor in CRC. Collectively, these observations highlight the biologic significance of epigenomic profiling on patient-matched primary specimens and identify this microenvironment-acquired super-enhancer as an oncogenic driver in the setting of the inflamed tumor.

PI3K/AKT/mTOR signaling pathway is one of the most frequently dysregulated pathway in cancer development and the serine/threonine kinase AKT functions as the key node in this pathway to regulate multiple cellular and physiological processes. AKT inhibitors that competitively bind the ATP pocket have been evaluated in clinical trials and have a tolerable toxicity profile with greater efficacy for tumors with pathway mutations; however, many PI3K/AKT pathway mutant tumors remain resistant. Proteolysis targeting chimeras (PROTACs) are a powerful targeted protein degradation technology that hijacks the cellular ubiquitin-proteasome system to induce selective polyubiquitination and degradation of the target proteins. Using this technology, we designed and developed a library of novel small-molecules putative degraders to degrade AKT by recruiting either the cereblon (CRBN) or von Hippel-Lindau (VHL) E3 ligase, that have the ability to degrade AKT in cells and lower downstream signaling to various extents. Though extensive structure-activity relationship studies on various linkers, E3 ligase ligands, and AKT binding moieties, we identified a novel VHL-recruiting AKT degrader, MS21, and characterized it using a panel of 38 cancer cell lines with diverse genotypes and tissue origins. Our results suggest that efficient pharmacologic degradation of AKT phosphorylated on threonine 308 and serine 473 leads to selective inhibition of the growth of tumor cells with alterations of HER2, PIK3CA, PTEN, or AKT1. In these PI3K/PTEN pathway mutant lines, AKT degradation by MS21 was superior to AKT kinase inhibition for reducing cell growth and sustaining lower signaling over many days, and inhibited tumor cell growths through lowering the level of Aurora Kinase B, which we found to be an AKT substrate protein. AKT degradation but not kinase inhibition profoundly lowered Aurora kinase B (AURKB) protein, which is known to be essential for cell division, and induced G2/M arrest and hyperploidy. PI3K activated AKT phosphorylation of AURKB on threonine 73, which protected it from proteasome degradation. A mutant of AURKB (T73E) that mimics phosphorylation and blocks its degradation rescued cells from growth inhibition by MS21. In addition, resistance to MS21 was found to be associated with low levels of baseline AKT phosphorylation in cells as well as mutation of either KRAS or BRAF, and resistance to MS21 could be overcome by the combination treatment of a MEK inhibitor trametinib with MS21, which not only inhibited MEK but also increased AKT phosphorylation and enhanced AKT degradation. Pan-cancer analysis identified that 19% of cases have PI3K/PTEN pathway mutation without RAS pathway mutation, suggesting that these cancer patients could benefit from AKT degrader therapy that leads to loss of AURKB. Citation Format: Jia Xu, Xufen Yu, Tiphaine C. Martin, Ankita Bansal, Kakit Cheung, Abigail Lubin, Elias Stratikopoulos, Kaitlyn M. Cahuzac, Li Wang, Ling Xie, Royce Zhou, Yudao Shen, Xuewei Wu, Shen Yao, Ruifang Qiao, Poulikos I. Poulikakos, Xian Chen, Jing Liu, Jian Jin, Ramon Parsons. AKT degradation selectively inhibits the growth of PI3K/PTEN pathway mutant cancers with wild type KRAS and BRAF by destabilizing Aurora kinase B [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB195.

Since the onset of COVID-19 pandemic, officials and several others have been making an effort to form informed decisions and take relevant measures to curb the outbreak. Mostly authorities have used standard statistical models, and epidemiological models to determine the outbreak. Although these models have shown to have accuracy in the past, they seem to be highly ineffective during the COVID-19 pandemic, mostly because of the complexities in it's outbreak. With the lack of studies done on the outbreak, it's imperative for us to determine the attributes which could show correlation with the spread for us to improve upon our predictions. Therefore in this paper the authors have tried to find the attributes which best help in the outbreak modelling and have applied those attributes to the traditional regression models to study their effects on the predictions.

Background: Non-Functional Requirements (NFR) have a direct impact on the architecture of the system, thus it is essential to identify NFRs in the initial phases of software development. Aim: The work is based on extraction of relevant keywords from NFR descriptions by employing text mining steps and thereafter classifying these descriptions into one of the nine types of NFRs. Method: For each NFR type, keywords are extracted from a set of pre-categorized specifications using Information-Gain measure. Then models using 8 Machine Learning (ML) techniques are developed for classification of NFR descriptions. A set of 15 projects (containing 326 NFR descriptions) developed by MS students at DePaul University are used to evaluate the models. Results: The study analyzes the performance of ML models in terms of classification and misclassification rate to determine the best model for predicting each type NFR descriptions. The Naïve Bayes model has performed best in predicting “maintainability” and “availability” type of NFRs. Conclusion: The NFR descriptions should be analyzed and mapped into their corresponding NFR types during the initial phases. The authors conducted cost benefit analysis to appreciate the advantage of using the proposed models.

Objective Clear cell renal cell carcinoma (ccRCC) is a subtype of kidney cancer defined by robust lipid accumulation, which prior studies have indicated plays an important role in tumor progression. We hypothesized that the peroxisome proliferator-activated receptor gamma (PPARγ), detected in both ccRCC tumors and cell lines, promotes lipid storage in ccRCC and contributes to tumorigenesis in this setting. PPARγ transcriptionally regulates a number of genes involved in lipid and glucose metabolism in adipocytes, yet its role in ccRCC has not been described. The objective of this study was to elucidate endogenous PPARγ function in ccRCC cells. Methods and results Using chromatin immunoprecipitation followed by deep sequencing (ChIP-seq), we found that PPARγ and its heterodimer RXR occupy the canonical DR1 PPAR binding motif at approximately 1000 locations throughout the genome that can be subdivided into adipose-shared and ccRCC-specific sites. CRISPR-Cas9 mediated, loss-of-function studies determined that PPARγ is dispensable for viability, proliferation, and migration of ccRCC cells in vitro and in vivo. Also, surprisingly, PPARγ deletion had little effect on the robust lipid accumulation that typifies the “clear cell” phenotype of kidney cancer. Conclusion Our results suggest that PPARγ plays neither a tumor suppressive nor oncogenic role in advanced ccRCC, and thus single-agent therapeutics targeting PPARγ are unlikely to be effective for the treatment of this disease. The unique cistrome of PPARγ in ccRCC cells demonstrates the importance of cell type in determining the functions of PPARγ., Highlights • PPARγ expression is elevated in ccRCC patient samples and cell lines relative to renal epithelium. • The PPARγ-RXR cistrome is distinct between ccRCC and adipocytes. • PPARγ is dispensable for viability, proliferation, and migration of ccRCC cells in vitro and in vivo. • Lipid storage and triglyceride synthesis occur independently of PPARγ in ccRCC.

Microbial desalination cell (MDC) is a propitious technology towards water desalination by utilizing wastewater as an energy source. In this study, a multi-chambered MDC was used to bioremediate steel plant wastewater using the same wastewater as a fuel for anodic bacteria. A pure culture of Pseudomonas putida MTCC 1194 was isolated and inoculated to remove toxic phenol. Three different inoculum conditions, namely P. putida (INC-A), a mixture of P. putida and activated sludge (INC-B), and activated sludge alone (INC-C) were employed in an anodic chamber to mainly compare the electricity generation and phenol degradation in MDCs. The study revealed the maximum phenol removal of 82 ± 2.4%, total dissolved solids (TDS) removal of 68 ± 1.5%, and power generation of 10.2 mW/m2 using INC-B. The synergistic interactions between microorganisms, can enhance the toxic phenol degradation and also electricity generation in MDC for onsite wastewater application.

CAPTCHA stands for Completely Automated Public Turing Test to Tell Computers and Humans Apart. CAPTCHAs are used as security mechanism in web applications to differentiate between real users and automated users, also known as bots. Text-based CAPTCHAs are the popularly used CAPTCHA schemes due to their simplicity and thus, they are still being used despite the proposal of several attack mechanisms. In this work, the authors have proposed a novel approach to solve CAPTCHA schemes. In this approach, the authors have used Depth First Search algorithm for the extraction of characters from CAPTCHAs and Convolutional Neural Network for recognizing these extracted characters. The proposed approach was validated on 3000+ CAPTCHA schemes and proved to be efficient by providing an average accuracy of more than 92.0% in detecting CAPTCHA schemes.

Ovarian torsion is a common diagnostic challenge constituting 2.7% to 7.4% of all gynaecological emergencies. It commonly occurs in women of reproductive age however pre-pubertal girls and postmenopausal women can also be affected. Ovarian torsion accounts for approximately 3% of all cases of children with acute abdominal pain and requires immediate surgical intervention. Other common acute adnexal pathologies include simple ovarian cysts (OCs) with or without rupture. Owing to the nonspecific clinical presentation and poor specificity of radiologic tests, the diagnosis of OT in girls remains challenging. Here we are reporting a case of acute abdomen with large cyst on ultrasound clinically presented as torsion later on in the post-operative period diagnosed with acute pancreatitis.

Bansal and Simon discuss strategies to block glutathione synthesis and utilization pathways to inhibit tumor propagation and treatment resistance., Glutathione (GSH) is the most abundant antioxidant found in living organisms and has multiple functions, most of which maintain cellular redox homeostasis. GSH preserves sufficient levels of cysteine and detoxifies xenobiotics while also conferring therapeutic resistance to cancer cells. However, GSH metabolism plays both beneficial and pathogenic roles in a variety of malignancies. It is crucial to the removal and detoxification of carcinogens, and alterations in this pathway can have a profound effect on cell survival. Excess GSH promotes tumor progression, where elevated levels correlate with increased metastasis. In this review, we discuss recent studies that focus on deciphering the role of GSH in tumor initiation and progression as well as mechanisms underlying how GSH imparts treatment resistance to growing cancers. Targeting GSH synthesis/utilization therefore represents a potential means of rendering tumor cells more susceptible to different treatment options such as chemotherapy and radiotherapy.

There are numerous reasons leading to change in software such as changing requirements, changing technology, increasing customer demands, fixing of defects etc. Thus, identifying and analyzing the change-prone classes of the software during software evolution is gaining wide importance in the field of software engineering. This would help software developers to judiciously allocate the resources used for testing and maintenance. Software metrics can be used for constructing various classification models which can be used for timely identification of change prone classes. Search based algorithms which form a subset of machine learning algorithms can be utilized for constructing prediction models to identify change prone classes of software. Search based algorithms use a fitness function to find the best optimal solution among all the possible solutions. In this work, we analyze the effectiveness of hybridized search based algorithms for change prediction. In other words, the aim of this work is to find whether search based algorithms are capable for accurate model construction to predict change prone classes. We have also constructed models using machine learning techniques and compared the performance of these models with the models constructed using Search Based Algorithms. The validation is carried out on two open source Apache projects, Rave and Commons Math. The results prove the effectiveness of hybridized search based algorithms in predicting change prone classes of software. Thus, they can be utilized by the software developers to produce an efficient and better developed software. Used hybridized search based algorithms to identify change prone classes.For empirical validation, two open source projects (Apache Rave,Commons Math) used.Assessed performance of search based algorithms using g-mean and accuracy.Machine learning models constructed and performance compared with hybridised models.Results showed that hybridised models outperformed machine learning models.

During the peptidoglycan (PG) maturation of mycobacteria, the glycan strands are interlinked by both 3-3 (between two meso -diaminopimelic acids [ meso -DAPs]) and 4-3 cross-links (between d -Ala and meso -DAP), though there is a predominance (60 to 80%) of 3-3 cross-links. The dd -carboxypeptidases ( dd -CPases) act on pentapeptides to generate tetrapeptides that are used by ld -transpeptidases as substrates to form 3-3 cross-links. Therefore, dd -CPases play a crucial role in mycobacterial PG cross-link formation. However, the physiology of dd -CPases in mycobacteria is relatively unexplored. In this study, we deleted two dd -CPase genes, msmeg_2433 and msmeg_2432 , both individually and in combination, from Mycobacterium smegmatis mc 2 155. Though the single dd -CPase gene deletions had no significant impact on the mycobacterial physiology, many interesting functional alterations were observed in the double-deletion mutant, viz ., a predominance in PG cross-link formation was shifted from 3-3 cross-links to 4-3, cell surface glycopeptidolipid (GPL) expression was reduced, and susceptibility to β-lactams and antitubercular agents was enhanced. Moreover, the survival rate of the double mutant within murine macrophages was higher than that of the parent. Interestingly, the complementation with any one of the dd -CPase genes could restore the wild-type phenotype. In a nutshell, we infer that the altered ratio of 4-3 to 3-3 PG cross-links might have influenced the expression of surface GPLs, colony morphology, biofilm formation, drug susceptibility, and subsistence of the cells within macrophages. IMPORTANCE The glycan strands in mycobacterial peptidoglycan (PG) are interlinked by both 3-3 and 4-3 cross-links. The dd -CPases generate tetrapeptides by acting on the pentapeptides, and ld -transpeptidases use tetrapeptides as substrates to form 3-3 cross-links. In this study, we showed that simultaneous deletions of two dd -CPases alter the nature of PG cross-linking from 3-3 cross-links to 4-3 cross-links. The deletions subsequently decrease the expression of glycopeptidolipids (significant surface lipid present in many nontuberculous mycobacteria, including Mycobacterium smegmatis ) and affect other physiological parameters, like cell morphology, growth rate, biofilm formation, antibiotic susceptibility, and survival within murine macrophages. Thus, unraveling the physiology of dd -CPases might help us design antimycobacterial therapeutics in the future.

DD-carboxypeptidases (DD-CPases) are low-molecular-mass (LMM) penicillin-binding proteins (PBPs) that are mainly involved in peptidoglycan remodelling, but little is known about the dd-CPases of mycobacteria. In this study, a putative DD-CPase of Mycobacterium smegmatis, MSMEG_2433 is characterized. The gene for the membrane-bound form of MSMEG_2433 was cloned and expressed in Escherichia coli in its active form, as revealed by its ability to bind to the Bocillin-FL (fluorescent penicillin). Interestingly, in vivo expression of MSMEG_2433 could restore the cell shape oddities of the septuple PBP mutant of E. coli, which was a prominent physiological characteristic of DD-CPases. Moreover, expression of MSMEG_2433 in trans elevated beta-lactam resistance in PBP deletion mutants (ΔdacAdacC) of E. coli, strengthening its physiology as a dd-CPase. To confirm the biochemical reason behind such physiological behaviours, a soluble form of MSMEG_2433 (sMSMEG_2433) was created, expressed and purified. In agreement with the observed physiological phenomena, sMSMEG_2433 exhibited DD-CPase activity against artificial and peptidoglycan-mimetic DD-CPase substrates. To our surprise, enzymic analyses of MSMEG_2433 revealed efficient deacylation for beta-lactam substrates at physiological pH, which is a unique characteristic of beta-lactamases. In addition to the MSMEG_2433 active site that favours dd-CPase activity, in silico analyses also predicted the presence of an omega-loop-like region in MSMEG_2433, which is an important determinant of its beta-lactamase activity. Based on the in vitro, in vivo and in silico studies, we conclude that MSMEG_2433 is a dual enzyme, possessing both DD-CPase and beta-lactamase activities.

Algae are a diverse group of photosynthetic organisms with profound bioremediation potential and industrial applications that can reduce the cost of expenditure on energy and fuels. Algae have been applied to various applications from fuels and medicines to wastewater treatment. Potential applications of phycoremediation have instigated laboratories to reinforce the development of algal-based technologies for better exploitation of their bioremediation potential and by-product generation ability. Many species of algae have been studied and utilized for their function in wastewater treatment. Certain technological drawbacks need to be addressed to manage the shortcomings of algae for industrial usage. This review mainly focuses on the state of the art applications of phycotechnology for wastewater remediation. Many problems related to algal technology have also been exemplified and discussed such as contamination, decreasing the resilience time and increasing the biomass of the algal culture. Possible solutions of these bottlenecks have been suggested to better apply phycotechnology for wastewater remediation in industries considering the environmental issues. Recently, the concept of genetic engineering is found very useful that has increased the potential of phycotechnology reducing the resilience time considerably. On the contrary, the negative impact of the algal-based technologies on the environment and economy has also been deliberated in order to effectively utilize and manage this important organism with utmost benefit. Lastly, the review illuminates the scope and options of future research in the area of phycoremediation technologies.

Background: Oral health is an important component of general health, with dental caries affecting a person's ability to eat, speak, or socialize. Dental caries is one of the most prevalent diseases affecting human beings and persists till date as a challenge to the medical and dental profession in particular and the society in general. Aim: The study aims to assess the prevalence of dental caries among 12–15-years-old school-going children of government and private schools in Agra city, Uttar Pradesh, India. Materials and Methods: A crosssectional study was conducted among 12–15–years-old school-going children of government and private schools in Agra city, Uttar Pradesh, India. A total of 1600 school children were through a multistage random sampling procedure. The children were examined according to the World Health Organization criteria, 2013. Information regarding a patient's demographic profile was recorded using a self–administered questionnaire. Chi-square test was used to find the significance of study parameters. The level of significance (P value) was fixed at 0.05. Results: The present study reveals a prevalence of dental caries was almost of the same magnitude in the 12 and 13 years of age group, and it was found to be 28.6% and 28.9%, respectively among government and private school children. It was found that among 14 and 15 years of age groups, dental caries affected 22.9% and 19.6% of the study participants, respectively from both government and private school children. The results were found to be highly statistically significant between government and private school children regarding the number of carious teeth. Conclusion: The prevalence of dental caries was found to be the major public health problem among both the government and private school children of Agra city, which need immediate attention. Therefore, there is a need to develop preventive and promotional oral health strategies to combat this disease.

Software product quality can be enhanced significantly if we have a good knowledge and understanding of the potential faults therein. This paper describes a study to build predictive models to identify parts of the software that have high probability of occurrence of fault. We have considered the effect of thresholds of object-oriented metrics on fault proneness and built predictive models based on the threshold values of the metrics used. Prediction of fault prone classes in earlier phases of software development life cycle will help software developers in allocating the resources efficiently. In this paper, we have used a statistical model derived from logistic regression to calculate the threshold values of object oriented, Chidamber and Kemerer metrics. Thresholds help developers to alarm the classes that fall outside a specified risk level. In this way, using the threshold values, we can divide the classes into two levels of risk - low risk and high risk. We have shown threshold effects at various risk levels and validated the use of these thresholds on a public domain, proprietary dataset, KC1 obtained from NASA and two open source, Promise datasets, IVY and JEdit using various machine learning methods and data mining classifiers. Interproject validation has also been carried out on three different open source datasets, Ant and Tomcat and Sakura. This will provide practitioners and researchers with well formed theories and generalised results. The results concluded that the proposed threshold methodology works well for the projects of similar nature or having similar characteristics.

Cell wall impermeability and active efflux of drugs are among the primary reasons for drug resistance in mycobacteria. Efflux pumps are tripartite membrane localized transport proteins that expel drug molecules outside the cells. Several of such efflux pumps are annotated in mycobacteria, but few have been characterized, like MSMEG\_2991, a putative efflux pump permease of Mycobacterium smegmatis . To substantiate this, we overexpressed MSMEG\_2991 protein in Escherichia coli 2443. Expression of MSMEG\_2991 elevated the resistance towards structurally unrelated groups of antibiotics. An active antibiotic efflux pump nature of MSMEG\_2991 was revealed by assessing the acquisition of ciprofloxacin in the absence and presence of the efflux pump inhibitor, carbonyl cyanide m-chlorophenyl hydrazone, indicating the involvement of proton-motive force (pmf) during the efflux activity. MSMEG\_2991 expression elevated biofilm formation in E. coli by 4-fold, keeping parity to some of the earlier reported efflux pumps. In silico analysis suggested the presence of 12 transmembrane helices in MSMEG\_2991 resembling EmrD efflux pump of E. coli . Based on in vivo and in silico analyses, MSMEG_2991 may be designated as a pmf-mediated multidrug efflux pump protein that expels diverse groups of antibiotics and might as well be involved in the biofilm enhancement.

To review advancements of fluoride in dentistry, a search of 21 electronic databases and World Wide Web was conducted. Relevant journals were hand searched and further information was requested from authors. Inclusion criteria were a predefined hierarchy of evidence and objectives. Study validity was assessed with checklists. Two reviewers independently screened sources, extracted data, and assessed validity. Fluoride has become an important tool in preventive dentistry. Current research is focused on the development of strategies to improve fluoride efficacy. Fluoride therapy in the form of varnish, gel, mouth rinse, or toothpaste has been used extensively as a caries-preventive intervention for over three decades. The purpose of this review is to inform the reader about new research related to the use of fluoride for the prevention of dental caries.

Aging research has been very successful at identifying signaling pathways and evolutionarily conserved genes that extend lifespan with the assumption that an increase in lifespan will also increase healthspan. However, it is largely unknown whether we are extending the healthy time of life or simply prolonging a period of frailty with increased incidence of age-associated diseases. Here we use Caenorhabditis elegans, one of the premiere systems for lifespan studies, to determine whether lifespan and healthspan are intrinsically correlated. We conducted multiple cellular and organismal assays on wild type as well as four long-lived mutants (insulin/insulin-like growth factor-1, dietary restriction, protein translation, mitochondrial signaling) in a longitudinal manner to determine the health of the animals as they age. We find that some long-lived mutants performed better than wild type when measured chronologically (number of days). However, all long-lived mutants increased the proportion of time spent in a frail state. Together, these data suggest that lifespan can no longer be the sole parameter of interest and reveal the importance of evaluating multiple healthspan parameters for future studies on antiaging interventions.

Penicillin-binding protein 5 (PBP5), a dd-carboxypeptidase, maintains cell shape and intrinsic beta-lactam resistance in E. coli. A strain lacking PBP5 loses intrinsic beta-lactam resistance and simultaneous lack of two other PBPs results in aberrantly shaped cells. PBP5 expression in trans complements the shape and restores the lost beta-lactam resistance. PBP5 has an 'Ω-loop'-like region similar to that in class A beta-lactamases. It was previously predicted that Leu182 present in the 'Ω-like' loop of PBP5 corresponds to Glu166 in PER-1 beta-lactamase. Here, we studied the physiological and biochemical effects of the Leu182Glu mutation in PBP5. Upon overexpression in septuple PBP mutants, ~75 % of cells were abnormally shaped and intrinsic beta-lactam resistance maintenance was partially lost. Biochemically, the purified soluble mutated PBP5 (smPBP5) retained low acylation ability for penicillin. The turnover number of smPBP5 for artificial and peptidoglycan mimetic substrates was ~10-fold less than that of the wild-type. Superimposition of the active-site residues of smPBP5 on PBP5 revealed that perturbation in the orientating key residues may explain the low turnover numbers. Therefore, we establish the involvement of Leu182 in maintaining the physiological and biochemical behaviour of E. coli PBP5.

Background: Insulin/IGF-1 signaling plays a central role in longevity across phylogeny. In C. elegans, the forkhead box O (FOXO) transcription factor, DAF-16, is the primary target of insulin/IGF-1 signaling, and multiple isoforms of DAF-16 (a, b, and d/f) modulate lifespan, metabolism, dauer formation, and stress resistance. Thus far, across phylogeny modulation of mammalian FOXOs and DAF-16 have focused on post-translational regulation with little focus on transcriptional regulation. In C. elegans ,w e have previously shown that DAF-16d/f cooperates with DAF-16a to promote longevity. In this study, we generated transgenic strains expressing near-endogenous levels of either daf-16a or daf-16d/f, and examined temporal expression of the isoforms to further define how these isoforms contribute to lifespan regulation. Results: Here, we show that DAF-16a is sensitive both to changes in gene dosage and to alterations in the level of insulin/IGF-1 signaling. Interestingly, we find that as worms age, the intestinal expression of daf-16d/f but not daf-16a is dramatically upregulated at the level of transcription. Preventing this transcriptional upregulation shortens lifespan, indicating that transcriptional regulation of daf-16d/f promotes longevity. In an RNAi screen of transcriptional regulators, we identify elt-2 (GATA transcription factor) and swsn-1 (core subunit of SWI/SNF complex) as key modulators of daf-16d/f gene expression. ELT-2 and another GATA factor, ELT-4, promote longevity via both DAF-16a and DAF-16d/f while the components of SWI/SNF complex promote longevity specifically via DAF-16d/f. Conclusions: Our findings indicate that transcriptional control of C. elegans FOXO/daf-16 is an essential regulatory event. Considering the conservation of FOXO across species, our findings identify a new layer of FOXO regulation as a potential determinant of mammalian longevity and age-related diseases such as cancer and diabetes.

The insulin/IGF-1 signaling (IIS) pathway is a conserved regulator of longevity, development, and metabolism. In Caenorhabditis elegans IIS involves activation of DAF-2 (insulin/IGF-1 receptor tyrosine kinase), AGE-1 (PI 3-kinase), and additional downstream serine/threonine kinases that ultimately phosphorylate and negatively regulate the single FOXO transcription factor homolog DAF-16. Phosphatases help to maintain cellular signaling homeostasis by counterbalancing kinase activity. However, few phosphatases have been identified that negatively regulate the IIS pathway. Here we identify and characterize pdp-1 as a novel negative modulator of the IIS pathway. We show that PDP-1 regulates multiple outputs of IIS such as longevity, fat storage, and dauer diapause. In addition, PDP-1 promotes DAF-16 nuclear localization and transcriptional activity. Interestingly, genetic epistasis analyses place PDP-1 in the DAF-7/TGF-β signaling pathway, at the level of the R-SMAD proteins DAF-14 and DAF-8. Further investigation into how a component of TGF-β signaling affects multiple outputs of IIS/DAF-16, revealed extensive crosstalk between these two well-conserved signaling pathways. We find that PDP-1 modulates the expression of several insulin genes that are likely to feed into the IIS pathway to regulate DAF-16 activity. Importantly, dysregulation of IIS and TGF-β signaling has been implicated in diseases such as Type 2 Diabetes, obesity, and cancer. Our results may provide a new perspective in understanding of the regulation of these pathways under normal conditions and in the context of disease., Author Summary Cells in the body respond to a variety of on/off signals that are relayed in a defined spatial and temporal manner. These signals influence several processes such as growth, fat storage, and the repair of damaged molecules. As humans age, the onset of diseases such as Type 2 Diabetes, obesity, and cancer often results from an imbalance in the levels of on/off signals in the cell. The insulin/IGF-1 signaling pathway is an important regulator of longevity, development, and metabolism across phylogeny. While the protein kinases that activate this pathway have been well studied, less is known about the protein phosphatases that tune down the signals. The roundworm C. elegans has been an excellent model system to study the role of insulin/IGF-1 signaling in the aging process. Here, we identify a new phosphatase that negatively regulates the insulin/IGF-1 pathway to enhance longevity and stress-resistance. Interestingly, the phosphatase achieves this function by tuning down the activity of a conserved TGF-β pathway, a pathway important for development. By reducing TGF-β pathway activity, this phosphatase decreases expression of insulin molecules that may stimulate the insulin/IGF-1 pathway. Our studies not only unravel a new regulator of these pathways, but also point to how they are more linked than previously thought. Both insulin/IGF-1 and TGF-β signaling have been implicated in age-associated diseases, and understanding their connection will provide us with potential therapeutic avenues.

The nematode Caenorhabditis elegans has been employed as a model organism to study human obesity due to the conservation of the pathways that regulate energy metabolism. To assay for fat storage in C. elegans, a number of fat-soluble dyes have been employed including BODIPY, Nile Red, Oil Red O, and Sudan Black. However, dye-labeled assays produce results that often do not correlate with fat stores in C. elegans. An alternative label-free approach to analyze fat storage in C. elegans has recently been described with coherent anti-Stokes Raman scattering (CARS) microscopy. Here, we compare the performance of CARS microscopy with standard dye-labeled techniques and biochemical quantification to analyze fat storage in wild type C. elegans and with genetic mutations in the insulin/IGF-1 signaling pathway including the genes daf-2 (insulin/IGF-1 receptor), rict-1 (rictor) and sgk-1 (serum glucocorticoid kinase). CARS imaging provides a direct measure of fat storage with unprecedented details including total fat stores as well as the size, number, and lipid-chain unsaturation of individual lipid droplets. In addition, CARS/TPEF imaging reveals a neutral lipid species that resides in both the hypodermis and the intestinal cells and an autofluorescent organelle that resides exclusively in the intestinal cells. Importantly, coherent addition of the CARS fields from the C-H abundant neutral lipid permits selective CARS imaging of the fat store, and further coupling of spontaneous Raman analysis provides unprecedented details including lipid-chain unsaturation of individual lipid droplets. We observe that although daf-2, rict-1, and sgk-1 mutants affect insulin/IGF-1 signaling, they exhibit vastly different phenotypes in terms of neutral lipid and autofluorescent species. We find that CARS imaging gives quantification similar to standard biochemical triglyceride quantification. Further, we independently confirm that feeding worms with vital dyes does not lead to the staining of fat stores, but rather the sequestration of dyes in lysosome-related organelles. In contrast, fixative staining methods provide reproducible data but are prone to errors due to the interference of autofluorescent species and the non-specific staining of cellular structures other than fat stores. Importantly, both growth conditions and developmental stage should be considered when comparing methods of C. elegans lipid storage. Taken together, we confirm that CARS microscopy provides a direct, non-invasive, and label-free means to quantitatively analyze fat storage in living C. elegans.

Due to various reasons such as ever increasing demands of the customer or change in the environment or detection of a bug, changes are incorporated in a software. This results in multiple versions or evolving nature of a software. Identification of parts of a software that are more prone to changes than others is one of the important activities. Identifying change prone classes will help developers to take focused and timely preventive actions on the classes of the software with similar characteristics in the future releases. In this paper, we have studied the relationship between various object oriented (OO) metrics and change proneness. We collected a set of OO metrics and change data of each class that appeared in two versions of an open source dataset, 'Java TreeView', i.e., version 1.1.6 and version 1.0.3. Besides this, we have also predicted various models that can be used to identify change prone classes, using machine learning and statistical techniques and then compared their performance. The results are analyzed using Area Under the Curve (AUC) obtained from Receiver Operating Characteristics (ROC) analysis. The results show that the models predicted using both machine learning and statistical methods demonstrate good performance in terms of predicting change prone classes. Based on the results, it is reasonable to claim that quality models have a significant relevance with OO metrics and hence can be used by researchers for early prediction of change prone classes.