Promoter Distortion and Opening in the RNA Polymerase II Cleft.

Transcription initiation requires opening of promoter DNA in the RNA polymerase II (Pol II) pre-initiation complex (PIC), but it remains unclear how this is achieved. Here we report the cryo-electron microscopic (cryo-EM) structure of a yeast PIC that contains underwound, distorted promoter DNA in the closed Pol II cleft. The DNA duplex axis is offset at the upstream edge of the initially melted DNA region (IMR) where DNA opening begins. Unstable IMRs are found in a subset of yeast promoters that we show can still initiate transcription after depletion of the transcription factor (TF) IIH (TFIIH) translocase Ssl2 (XPB in human) from the nucleus in vivo. PIC-induced DNA distortions may thus prime the IMR for melting and may explain how unstable IMRs that are predicted in promoters of Pol I and Pol III can open spontaneously. These results suggest that DNA distortion in the polymerase cleft is a general mechanism that contributes to promoter opening.

A Performance Analysis of Security Protocols for Distributed Measurement Systems Based on Internet of Things with Constrained Hardware and Open Source Infrastructures.

The widespread adoption of Internet of Things (IoT) devices in home, industrial, and business environments has made available the deployment of innovative distributed measurement systems (DMS). This paper takes into account constrained hardware and a security-oriented virtual local area network (VLAN) approach that utilizes local message queuing telemetry transport (MQTT) brokers, transport layer security (TLS) tunnels for local sensor data, and secure socket layer (SSL) tunnels to transmit TLS-encrypted data to a cloud-based central broker. On the other hand, the recent literature has shown a correlated exponential increase in cyber attacks, mainly devoted to destroying critical infrastructure and creating hazards or retrieving sensitive data about individuals, industrial or business companies, and many other entities. Much progress has been made to develop security protocols and guarantee quality of service (QoS), but they are prone to reducing the network throughput. From a measurement science perspective, lower throughput can lead to a reduced frequency with which the phenomena can be observed, generating, again, misevaluation. This paper does not give a new approach to protect measurement data but tests the network performance of the typically used ones that can run on constrained hardware. This is a more general scenario typical for IoT-based DMS. The proposal takes into account a security-oriented VLAN approach for hardware-constrained solutions. Since it is a worst-case scenario, this permits the generalization of the achieved results. In particular, in the paper, all OpenSSL cipher suites are considered for compatibility with the Mosquitto server. The most used key metrics are evaluated for each cipher suite and QoS level, such as the total ratio, total runtime, average runtime, message time, average bandwidth, and total bandwidth. Numerical and experimental results confirm the proposal's effectiveness in foreseeing the minimum network throughput concerning the selected QoS and security. Operating systems yield diverse performance metric values based on various configurations. The primary objective is identifying algorithms to ensure suitable data transmission and encryption ratios. Another aim is to explore algorithms that ensure wider compatibility with existing infrastructures supporting MQTT technology, facilitating secure connections for geographically dispersed DMS IoT networks, particularly in challenging environments like suburban or rural areas. Additionally, leveraging open firmware on constrained devices compatible with various MQTT protocols enables the customization of the software components, a crucial necessity for DMS.

Systematic mutagenesis of TFIIH subunit p52/Tfb2 identifies residues required for XPB/Ssl2 subunit function and genetic interactions with TFB6.

TFIIH is an evolutionarily conserved complex that plays central roles in both RNA polymerase II (pol II) transcription and DNA repair. As an integral component of the pol II preinitiation complex, TFIIH regulates pol II enzyme activity in numerous ways. The TFIIH subunit XPB/Ssl2 is an ATP-dependent DNA translocase that stimulates promoter opening prior to transcription initiation. Crosslinking-mass spectrometry and cryo-EM results have shown a conserved interaction network involving XPB/Ssl2 and the C-terminal Hub region of the TFIIH p52/Tfb2 subunit, but the functional significance of specific residues is unclear. Here, we systematically mutagenized the HubA region of Tfb2 and screened for growth phenotypes in a TFB6 deletion background in Saccharomyces cerevisiae. We identified six lethal and 12 conditional mutants. Slow growth phenotypes of all but three conditional mutants were relieved in the presence of TFB6, thus identifying a functional interaction between Tfb2 HubA mutants and Tfb6, a protein that dissociates Ssl2 from TFIIH. Our biochemical analysis of Tfb2 mutants with severe growth phenotypes revealed defects in Ssl2 association, with similar results in human cells. Further characterization of these tfb2 mutant cells revealed defects in GAL gene induction, and reduced occupancy of TFIIH and pol II at GAL gene promoters, suggesting that functionally competent TFIIH is required for proper pol II recruitment to preinitiation complexes in vivo. Consistent with recent structural models of TFIIH, our results identify key residues in the p52/Tfb2 HubA domain that are required for stable incorporation of XPB/Ssl2 into TFIIH and for pol II transcription.

Staphylococcal superantigen-like protein 10 enhances the amyloidogenic biofilm formation in Staphylococcus aureus.

Staphylococcus aureus is a highly infectious pathogen that represents a significant burden on the current healthcare system. Bacterial attachment to medical implants and host tissue, and the establishment of a mature biofilm, play an important role in chronic diseases such as endocarditis, osteomyelitis and wound infections. These biofilms decrease bacterial susceptibility to antibiotics and immune defences, making the infections challenging to treatment. S. aureus produces numerous exotoxins that contribute to the pathogenesis of the bacteria. In this study, we have identified a novel function of staphylococcal superantigen-like protein 10 (SSL10) in enhancing the formation of staphylococcal biofilms. Biofilm biomass is significantly increased when SSL10 is added exogenously to bacterial cultures, whereas SSL2 and SSL12 are found to be less active. Exogenously added SSL10 mask the surface charge of the bacterial cells and lowers their zeta potential, leading to the aggregation of the cells. Moreover, the biofilm formation by SSL10 is governed by amyloid aggregation, as evident from spectroscopic and microscopic studies. These findings thereby give the first overview of the SSL-mediated amyloid-based biofilm formation and further drive the future research in identifying potential molecules for developing new antibacterial therapies against Staphylococcus aureus.

Luminescence Studies on Dy3+ Doped Calcium Aluminum Borosilicate (CABS) Glasses for White Light Emission and Applications in w-LEDs.

Dy3+ doped calcium aluminum borosilicate (CABS) glasses have been synthesized via quick melt quench technique. CABS: xDy3+ glasses (x = 0.1, 0.5, 1, 1.5 and 2 mol%) were subjected to various morphological and photoluminescence studies. X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy were conducted to study the structural and bonding nature of the undoped glass. The excitation spectra of Dy3+ doped CABS glasses under 574 nm emission show many sharp peaks amongst which the transition from 6H15/2 → 6P7/2 (351 nm) had the highest intensity. Under 351 nm excitation, glasses exhibit sharp peaks in the blue, yellow and red regions corresponding to the transitions 4F9/2 → 6H15/2, 6H13/2, 6H11/2 and 6H9/2 respectively. The dipole-dipole nature of the interaction between the Dy3+ ions is confirmed via Dexter theory and Inokuti-Hirayama (I-H) model. CIE coordinates estimated from the emission profiles of these glasses under 351 nm excitation fall in the white region. Considering that these glasses exhibit sharp visible emission under UV excitation, have stable yellow to blue (Y/B) ratios and fast decays with intense energy transfers, we propose to utilise these glasses for white light generation and other white light LED (w-LED) and solid-state lighting (SSL) applications.

Design of Self-Integrating Transient Surface Current Density Sensor Integrated Fiber Transmission Link.

The transient surface current density reflects the external coupling of the electromagnetic pulse (EMP) to the tested device. In this paper, the generation mechanism and measurement principle of conductor surface current density are introduced, and the surface current density distribution irradiated by EMP on a typical aircraft structure is simulated and analyzed. The traditional surface current density is usually measured by B-dot antenna, but its output signal is the differential of the measured signal, so additional integrators or numerical integration of the measured data are required. In this paper, a self-integrating surface current sensor based on optical fiber transmission is designed based on the shielded loop antenna with gap structure. The output signal is the real signal waveform to be measured. Compared with coaxial cables, integrated optical fiber transmission improves the anti-interference ability of long-distance transmission signals. At the same time, the design process of the sensor is introduced in detail. The bandwidth of the sensor is 300 kHz~500 MHz, the sensitivity is calibrated at 1.23 (A·m-1)/mV, and the dynamic range is ±25~1400 A·m-1 (35 dB). The surface current of a metal plate is measured in a bounded wave electromagnetic pulse simulator using a detector developed in this paper. The test results show that the developed sensor has good engineering applicability.

ACG-EmoCluster: A Novel Framework to Capture Spatial and Temporal Information from Emotional Speech Enhanced by DeepCluster.

Speech emotion recognition (SER) is a task that tailors a matching function between the speech features and the emotion labels. Speech data have higher information saturation than images and stronger temporal coherence than text. This makes entirely and effectively learning speech features challenging when using feature extractors designed for images or texts. In this paper, we propose a novel semi-supervised framework for extracting spatial and temporal features from speech, called the ACG-EmoCluster. This framework is equipped with a feature extractor for simultaneously extracting the spatial and temporal features, as well as a clustering classifier for enhancing the speech representations through unsupervised learning. Specifically, the feature extractor combines an Attn-Convolution neural network and a Bidirectional Gated Recurrent Unit (BiGRU). The Attn-Convolution network enjoys a global spatial receptive field and can be generalized to the convolution block of any neural networks according to the data scale. The BiGRU is conducive to learning temporal information on a small-scale dataset, thereby alleviating data dependence. The experimental results on the MSP-Podcast demonstrate that our ACG-EmoCluster can capture effective speech representation and outperform all baselines in both supervised and semi-supervised SER tasks.

Semi-Supervised Anomaly Detection of Dissolved Oxygen Sensor in Wastewater Treatment Plants.

As the world progresses toward a digitally connected and sustainable future, the integration of semi-supervised anomaly detection in wastewater treatment processes (WWTPs) promises to become an essential tool in preserving water resources and assuring the continuous effectiveness of plants. When these complex and dynamic systems are coupled with limited historical anomaly data or complex anomalies, it is crucial to have powerful tools capable of detecting subtle deviations from normal behavior to enable the early detection of equipment malfunctions. To address this challenge, in this study, we analyzed five semi-supervised machine learning techniques (SSLs) such as Isolation Forest (IF), Local Outlier Factor (LOF), One-Class Support Vector Machine (OCSVM), Multilayer Perceptron Autoencoder (MLP-AE), and Convolutional Autoencoder (Conv-AE) for detecting different anomalies (complete, concurrent, and complex) of the Dissolved Oxygen (DO) sensor and aeration valve in the WWTP. The best results are obtained in the case of Conv-AE algorithm, with an accuracy of 98.36 for complete faults, 97.81% for concurrent faults, and 98.64% for complex faults (a combination of incipient and concurrent faults). Additionally, we developed an anomaly detection system for the most effective semi-supervised technique, which can provide the detection of delay time and generate a fault alarm for each considered anomaly.

Genome-Wide Analysis of Strictosidine Synthase-like Gene Family Revealed Their Response to Biotic/Abiotic Stress in Poplar.

The strictosidine synthase-like (SSL) gene family is a small plant immune-regulated gene family that plays a critical role in plant resistance to biotic/abiotic stresses. To date, very little has been reported on the SSL gene in plants. In this study, a total of thirteen SSLs genes were identified from poplar, and these were classified into four subgroups based on multiple sequence alignment and phylogenetic tree analysis, and members of the same subgroup were found to have similar gene structures and motifs. The results of the collinearity analysis showed that poplar SSLs had more collinear genes in the woody plants Salix purpurea and Eucalyptus grandis. The promoter analysis revealed that the promoter region of PtrSSLs contains a large number of biotic/abiotic stress response elements. Subsequently, we examined the expression patterns of PtrSSLs following drought, salt, and leaf blight stress, using RT-qPCR to validate the response of PtrSSLs to biotic/abiotic stresses. In addition, the prediction of transcription factor (TF) regulatory networks identified several TFs, such as ATMYB46, ATMYB15, AGL20, STOP1, ATWRKY65, and so on, that may be induced in the expression of PtrSSLs in response to adversity stress. In conclusion, this study provides a solid basis for a functional analysis of the SSL gene family in response to biotic/abiotic stresses in poplar.

Interaction of ßL- and γ-Crystallin with Phospholipid Membrane Using Atomic Force Microscopy.

Highly concentrated lens proteins, mostly ß- and γ-crystallin, are responsible for maintaining the structure and refractivity of the eye lens. However, with aging and cataract formation, ß- and γ-crystallin are associated with the lens membrane or other lens proteins forming high-molecular-weight proteins, which further associate with the lens membrane, leading to light scattering and cataract development. The mechanism by which ß- and γ-crystallin are associated with the lens membrane is unknown. This work aims to study the interaction of ß- and γ-crystallin with the phospholipid membrane with and without cholesterol (Chol) with the overall goal of understanding the role of phospholipid and Chol in ß- and γ-crystallin association with the membrane. Small unilamellar vesicles made of Chol/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (Chol/POPC) membranes with varying Chol content were prepared using the rapid solvent exchange method followed by probe tip sonication and then dispensed on freshly cleaved mica disk to prepare a supported lipid membrane. The ßL- and γ-crystallin from the cortex of the bovine lens was used to investigate the time-dependent association of ßL- and γ-crystallin with the membrane by obtaining the topographical images using atomic force microscopy. Our study showed that ßL-crystallin formed semi-transmembrane defects, whereas γ-crystallin formed transmembrane defects on the phospholipid membrane. The size of semi-transmembrane defects increases significantly with incubation time when ßL-crystallin interacts with the membrane. In contrast, no significant increase in transmembrane defect size was observed in the case of γ-crystallin. Our result shows that Chol inhibits the formation of membrane defects when ßL- and γ-crystallin interact with the Chol/POPC membrane, where the degree of inhibition depends upon the amount of Chol content in the membrane. At a Chol/POPC mixing ratio of 0.3, membrane defects were observed when both ßL- and γ-crystallin interacted with the membrane. However, at a Chol/POPC mixing ratio of 1, no association of γ-crystallin with the membrane was observed, which resulted in a defect-free membrane, and the severity of the membrane defect was decreased when ßL-crystallin interacted with the membrane. The semi-transmembrane or transmembrane defects formed by the interaction of ßL- and γ-crystallin on phospholipid membrane might be responsible for light scattering and cataract formation. However, Chol suppressed the formation of such defects in the membrane, likely maintaining lens membrane homeostasis and protecting against cataract formation.

Binding of ßL-Crystallin with Models of Animal and Human Eye Lens-Lipid Membrane.

Several discoveries show that with age and cataract formation, ß-crystallin binds with the lens membrane or associates with other lens proteins, which bind with the fiber cell plasma membrane, accompanied by light scattering and cataract formation. However, how lipids (phospholipids and sphingolipids) and cholesterol (Chol) influence ß-crystallin binding to the membrane is unclear. This research aims to elucidate the role of lipids and Chol in the binding of ß-crystallin to the membrane and the membrane's physical properties (mobility, order, and hydrophobicity) with ß-crystallin binding. We used electron paramagnetic resonance (EPR) spin-labeling methods to investigate the binding of ßL-crystallin with a model of porcine lens-lipid (MPLL), model of mouse lens-lipid (MMLL), and model of human lens-lipid (MHLL) membrane with and without Chol. Our results show that ßL-crystallin binds with all of the investigated membranes in a saturation manner, and the maximum parentage of the membrane surface occupied (MMSO) by ßL-crystallin and the binding affinity (Ka) of ßL-crystallin to the membranes followed trends: MMSO (MPLL) > MMSO (MMLL) > MMSO (MHLL) and Ka (MHLL) > Ka (MMLL) ≈ Ka (MPLL), respectively, in which the presence of Chol reduces the MMSO and Ka for all membranes. The mobility near the headgroup regions of the membranes decreases with an increase in the binding of ßL-crystallin; however, the decrease is more pronounced in the MPLL and MMLL membranes than the MHLL membrane. In the MPLL and MMLL membranes, the membranes become slightly ordered near the headgroup with an increase in ßL-crystallin binding compared to the MHLL membrane. The hydrophobicity near the headgroup region of the membrane increases with ßL-crystallin binding; however, the increase is more pronounced in the MPLL and MMLL membranes than the MHLL membrane, indicating that ßL-crystallin binding creates a hydrophobic barrier for the passage of polar molecules, which supports the barrier hypothesis in cataract formation. However, in the presence of Chol, there is no significant increase in hydrophobicity with ßL-crystallin binding, suggesting that Chol prevents the formation of a hydrophobic barrier, possibly protecting against cataract formation.

Disparate age and sex distribution of sessile serrated lesions and conventional adenomas in an outpatient colonoscopy population-implications for colorectal cancer screening?

PURPOSE: Colorectal cancer (CRC) is increasingly diagnosed in individuals aged < 50 years, resulting in advocacy of screening from age 45 years. Despite existing knowledge associating CRC with conventional adenomas, the significance of sessile serrated lesions (SSLs) on the burden of CRC is less detailed. We aimed to provide contemporary estimates for SSL prevalence and examine patient and procedure factors associated with SSL detection. METHODS: Retrospective observational study examining associations between SSL and conventional adenoma detection, polyp histopathology, patient, and procedure characteristics in an outpatient colonoscopy unit over 12 months. RESULTS: From 2097 colonoscopies, SSL detection was 13.8% overall and 12.5% in patients < 50 years. SSLs were mostly proximal in location (64%), and SSL detection was significantly higher in females compared with males (16.2% vs. 11.7%, p = 0.003), particularly in those < 50 years (16.8% vs. 8.6%, p < 0.001). In multivariable analysis, SSL detection was associated with female sex (adjusted odds ratio [aOR] 1.48, 95% confidence interval [CI] 1.15-1.91), synchronous conventional adenoma detection (aOR 1.36, 95% CI 1.04-1.78) and BMI ≥ 25 kg/m2 (aOR 1.34, 95% CI 1.02-1.77). Conventional adenoma detection was 33.6% and associated with age ≥ 50 years (aOR 3.57, 95% CI 2.84-4.47) and synchronous SSL detection (aOR 1.36, 95% CI 1.03-1.79). CONCLUSIONS: We observed age and sex disparities in polyp types and prevalence in this outpatient colonoscopy population. SSLs were most prevalent in females aged < 50 years, suggesting a potential increased susceptibility of young females to SSLs and CRC. Our findings may have implications for the design of CRC screening programs.

Analysis of Texture and Color Enhancement Imaging for Improving the Visibility of Non-polypoid Colorectal Lesions.

INTRODUCTION: An endoscopic system using 5-color light-emitting diodes (LEDs) (EVIS X1; Olympus Co., Tokyo, Japan), which includes texture and color enhancement imaging (TXI), has been released. In this study, we analyzed the effects of TXI on the visibility of non-polypoid colorectal lesions and its diagnostic accuracy. METHODS: We reviewed 101 non-polypoid lesions from 26 patients observed with white light imaging (WLI), narrow band imaging (NBI), and TXI. One representative image of each mode was evaluated by 6 endoscopists using a polyp visibility score of 4 (excellent) to 1 (poor). We calculated the color difference (CD) values for each lesion in the three modes. For tumor characteristics, one representative image of TXI and NBI magnification was evaluated by 3 experts according to a NBI classification. RESULTS: The least squares means [95% confidence interval] of polyp visibility score of TXI (3.42 [3.06-3.77]) was significantly higher than that of WLI (2.85 [2.49-3.20], p < 0.001) but not that of NBI (3.33 [2.98-3.69], p = 0.258). The CD value of TXI (13.3 ± 6.3) was higher than that of WLI (9.7 ± 6.0, p < 0.001) but not that of NBI (13.1 ± 6.8, p = 0.81). For sessile serrated lesions, the CD value of TXI (11.1 ± 4.4) tended to be lower than that of NBI (12.6 ± 6.0, p = 0.07). The diagnostic accuracy and confidence level of magnification for NBI were significantly better than those for TXI (87.1 vs. 80.5%, p = 0.027, 87.5 vs. 62.7%, p < 0.001, respectively). CONCLUSION: TXI showed better visibility than WLI in terms of the endoscopist's score and CD value and may improve polyp detection.

Circumferential submucosal incision prior to endoscopic mucosal resection versus conventional endoscopic mucosal resection for colorectal lesions with endoscopic features of sessile serrated lesions.

BACKGROUND AND AIMS: Sessile serrated lesions (SSLs) are more prone to incomplete resection than conventional adenomas. This study evaluated whether circumferential submucosal incision prior to endoscopic mucosal resection (CSI-EMR) can increase the rate of complete and en bloc resections of colorectal lesions with endoscopic features of SSL. METHODS: Retrospective analyses and propensity score matching were performed for the resection of colorectal lesions ≥ 10 mm with endoscopic features of SSL. RESULTS: After 1:1 ratio matching, 127 lesions in the CSI-EMR group and 127 in the EMR group were selected for analysis. The median size of the lesions was 15 mm (IQR 12-16) in both groups. There was no significant difference in either the complete resection rate or en bloc resection rate between CSI-EMR and EMR groups (96.9% vs. 92.9%, P = 0.155; 92.1% vs. 89.0%, P = 0.391). By contrast, the R0 resection rate was significantly higher in the CSI-EMR group than in the EMR group (89.8% vs. 59.8%, P < 0.001). The median procedure time was significantly longer in the CSI-EMR group than in the EMR group (6.28 min vs. 2.55 min, P < 0.001), whereas there was no significant difference between the two groups in the incidence of adverse events or recurrence rate. Multivariate analysis showed that CSI-EMR was the only factor significantly associated with R0 resection (P < 0.001). CONCLUSIONS: For colorectal lesions with endoscopic features of SSL, CSI-EMR does not increase the complete or en bloc resection rate, but does increase the R0 resection rate. The procedure time is longer for CSI-EMR than EMR. The association of CSI-EMR with R0 resection and non-recurrence should be further evaluated.

Survey on Self-Supervised Learning: Auxiliary Pretext Tasks and Contrastive Learning Methods in Imaging.

Although deep learning algorithms have achieved significant progress in a variety of domains, they require costly annotations on huge datasets. Self-supervised learning (SSL) using unlabeled data has emerged as an alternative, as it eliminates manual annotation. To do this, SSL constructs feature representations using pretext tasks that operate without manual annotation, which allows models trained in these tasks to extract useful latent representations that later improve downstream tasks such as object classification and detection. The early methods of SSL are based on auxiliary pretext tasks as a way to learn representations using pseudo-labels, or labels that were created automatically based on the dataset's attributes. Furthermore, contrastive learning has also performed well in learning representations via SSL. To succeed, it pushes positive samples closer together, and negative ones further apart, in the latent space. This paper provides a comprehensive literature review of the top-performing SSL methods using auxiliary pretext and contrastive learning techniques. It details the motivation for this research, a general pipeline of SSL, the terminologies of the field, and provides an examination of pretext tasks and self-supervised methods. It also examines how self-supervised methods compare to supervised ones, and then discusses both further considerations and ongoing challenges faced by SSL.

Impediment effect of chemical agents (additives) on gluten development in cookie dough.

Most of the countries grow wheat varieties according to the product specificity while others lack such a system of special purpose wheat production. Those countries face problems in quality of bakery products like cookies/biscuits and breads. Cookie manufacturers require low protein to prepare good quality cookies. In case of high protein wheat flour, one solution is to hinder the gluten development during dough formation. It can be achieved by using chemical additives like Sodium Stearoyl 2-Lactylate (SSL), L-Cysteine (CYS) and Lecithin (LEC). So, current study was designed to add these additives in dough to hinder gluten development. For this purpose, wheat flour of two local mills was procured and analyzed for chemical & rheological traits. Furthermore, flour was used to prepare cookies with the addition of chemical additives and these cookies were evaluated for textural hardness, physical parameters, and sensorial characteristics. Results showed that wheat flours have optimum values of chemical & farinographic attributes. Chemical additives significantly affect diameter, thickness, spread factor, hardness, flavor, taste, and texture of cookies. Conclusively, SSL can be added in mixed wheat flour @ 0.5-1% to get cookies of good quality.

Genome sequencing and molecular characterisation of XDR Acinetobacter baumannii reveal complexities in resistance: Novel combination of sulbactam-durlobactam holds promise for therapeutic intervention.

Emerging nosocomial strains of Acinetobacter baumannii are of recent concern as they are expressing extensive drug resistance (XDR). Using whole-genome sequencing and molecular characterisation analysis, the current study reveals the presence of carbapenemase genes in 92.86% of studied Indian isolates. These included blaOXA-51 , blaOXA-23 , blaOXA-58 , and blaNDM genes, with over a third expressing dual carbapenemase genes. As per the MLST scheme, IC2Oxf /CC2Pas was the predominant clone, with 57.14% isolates belonging to this lineage. The presence of these carbapenemase genes resulted in sulbactam (SUL) resistance (MIC: 16-256 µg/ml) in all of the studied isolates. The efficacy of durlobactam (DUR), a novel ß-lactamase inhibitor that also inhibits PBP2 was assessed through in silico intermolecular interaction analysis. Several nonsynonymous single nucleotide polymorphisms were identified in PBP2 (G264S, I108V, S259T) and PBP3 (A515V, T526S) sequences. Minimal variations were recorded in the protein backbone dynamics in active-site motifs of wild-type and mutants, which correlated with negligible binding energy fluctuations for the PBP3-SUL (-5.85 ± 0.04 kcal/mol) and PBP2-DUR (-5.16 ± 0.66 kcal/mol) complexes. Furthermore, higher binding affinities and low inhibition constants were noted in OXA23-DUR (-7.36 kcal/mol; 4.01 µM), OXA58-DUR (-6.44 kcal/mol; 19.07 µM), and NDM-DUR (-6.82 kcal/mol; 10.01 µM) complexes when compared with the conventional drugs avibactam and aztreonam. Stable interaction profiles of DUR with carbapenemases can possibly restore SUL activity against both PBP3WT and PBP3MTs . The study establishes the efficacy of the novel SUL-DUR combination as a successful treatment strategy in combating emerging XDR strains of A. baumannii.

Usefulness of the Japan narrow-band imaging expert team classification system for the diagnosis of sessile serrated lesion with dysplasia/carcinoma.

BACKGROUND: Sessile serrated lesion (SSL) is a colorectal polyp that has malignant potential. However, the dysplastic components within an SSL can be difficult to diagnose with conventional endoscopy, because most SSLs with dysplasia/carcinoma have subtle mucosal features. Many studies have indicated that narrow-band imaging (NBI) observations of colorectal polyps are very useful, accurate predictors of histology. We aimed to verify the usefulness of the Japan NBI Expert Team (JNET) classification system for the diagnosis of SSLs with dysplasia/carcinoma. METHODS: We examined 709 endoscopically or surgically resected lesions that were pathologically diagnosed as SSL, including 647 with no dysplasia, 37 with low-grade dysplasia, 15 with high-grade dysplasia, and 10 with submucosal invasive carcinoma. We retrospectively evaluated their clinicopathologic characteristics and conventional endoscopic and magnifying NBI endoscopic findings using the JNET system. RESULTS: Cases in all groups were more frequently located in the proximal colon. Submucosal invasive carcinomas were significantly larger than no dysplasia and low-grade dysplasia lesions. Almost all studied lesions (96.3%) were covered with a mucus cap. Five hundred and eighty (81.8%) lesions exhibited dark spots inside the crypts, which are NBI findings' characteristic of SSL. As for the JNET classification of magnifying NBI endoscopic findings, all 709 lesions showed Type 1. Six hundred and eighteen (95.5%) SSLs with no dysplasia lesions exhibited Type 1 only, whereas 52 (83.9%) SSLs with dysplasia/carcinoma had a combination of Type 1 and Type 2A, 2B, or 3, corresponding to SSL and dysplasia/carcinoma, respectively. The JNET classification had high sensitivity (83.9%), specificity (95.5%), and overall diagnostic accuracy (94.5%) for diagnosing SSLs with dysplasia/carcinoma. CONCLUSIONS: Use of magnifying NBI endoscopy with the JNET classification might be useful for diagnosing SSLs with dysplasia/carcinoma. This increased awareness may also improve the recognition of SSLs with dysplasia/carcinoma.

Growth of wood-inhabiting yeasts of the Faroe Islands in the presence of spent sulphite liquor.

In the microbial community of decaying wood, yeasts are important for the recycling of nutrients. Nevertheless, information on their biodiversity in this niche in the Northern hemisphere is limited. Wood-colonising yeasts encounter identical and similar growth-inhibitory compounds as those in spent sulphite liquor (SSL), an energy-rich, acid hydrolysate and waste product from the paper industry, which may render them well-suited for cultivation in SSL. In the present study, yeasts were isolated from decaying wood on the Faroe Islands and identified based on sequence homology of the ITS and D1/D2 regions. Among the yeasts isolated, Candida argentea, Cystofilobasidium infirmominiatum, Naganishia albidosimilis, Naganishia onofrii, Holtermanniella takashimae and Goffeauzyma gastrica were new to decaying wood in cold and temperate climates. C. argentea and Rhodotorula are rarely-isolated species, with no previous documentation from cold and maritime climates. The isolates were further tested for growth in a medium with increasing concentrations of softwood SSL. Most grew in the presence of 10% SSL. Isolates of Debaryomyces sp., C. argentea and Rhodotorula sp. were the most tolerant. Representatives of Debaryomyces and Rhodotorula have previously been found in decaying wood. In contrast, the least tolerant isolates belonged to species that are rarely reported from decaying wood. The relative importance of individual inhibitors to yeast growth is discussed. To our knowledge, none of the present yeast species have previously been cultivated in SSL medium. Decaying wood can be a useful future source of yeasts for valorisation of various hydrolysates to industrial chemicals and biofuels.

Liposomes Targeting P21 Activated Kinase-1 (PAK-1) and Selective for Secretory Phospholipase A2 (sPLA2) Decrease Cell Viability and Induce Apoptosis in Metastatic Triple-Negative Breast Cancer Cells.

P21 activated kinases (or group I PAKs) are serine/threonine kinases whose expression is altered in prostate and breast cancers. PAK-1 activity is inhibited by the small molecule "Inhibitor targeting PAK-1 activation-3" (IPA-3), which has selectivity for PAK-1 but is metabolically unstable. Secretory Group IIA phospholipase A2 (sPLA2) expression correlates to increased metastasis and decreased survival in many cancers. We previously designed novel liposomal formulations targeting both PAK-1 and sPLA2, called Secretory Phospholipase Responsive liposomes or SPRL-IPA-3, and demonstrated their ability to alter prostate cancer growth. The efficacy of SPRL against other types of cancers is not well understood. We addressed this limitation by determining the ability of SPRL to induce cell death in a diverse panel of cells representing different stages of breast cancer, including the invasive but non-metastatic MCF-7 cells, and metastatic triple-negative breast cancer (TNBC) cells such as MDA-MB-231, MDA-MB-468, and MDA-MB-435. We investigated the role of sPLA2 in the disposition of these liposomes by comparing the efficacy of SPRL-IPA-3 to IPA-3 encapsulated in sterically stabilized liposomes (SSL-IPA-3), a formulation shown to be less sensitive to sPLA2. Both SSL-IPA-3 and SPRL-IPA-3 induced time- and dose-dependent decreases in MTT staining in all cell lines tested, but SPRL-IPA-3-induced effects in metastatic TNBC cell lines were superior over SSL-IPA-3. The reduction in MTT staining induced by SPRL-IPA-3 correlated to the expression of Group IIA sPLA2. sPLA2 expression also correlated to increased induction of apoptosis in TNBC cell lines by SPRL-IPA-3. These data suggest that SPRL-IPA-3 is selective for metastatic TNBC cells and that the efficacy of SPRL-IPA-3 is mediated, in part, by the expression of Group IIA sPLA2.