Molecular characterization of the PhoPQ-PmrD-PmrAB mediated pathway regulating polymyxin B resistance in Klebsiella pneumoniae CG43

<p>Abstract</p> <p>Background</p> <p>The cationic peptide antibiotic polymyxin has recently been reevaluated in the treatment of severe infections caused by gram negative bacteria.</p> <p>Methods</p> <p>In this study, the genetic determinants for capsular polysaccharide level and lipopolysaccharide modification involved in polymyxin B resistance of the opportunistic pathogen <it>Klebsiella pneumoniae </it>were characterized. The expressional control of the genes responsible for the resistance was assessed by a LacZ reporter system. The PmrD connector-mediated regulation for the expression of <it>pmr </it>genes involved in polymyxin B resistance was also demonstrated by DNA EMSA, two-hybrid analysis and <it>in vitro </it>phosphor-transfer assay.</p> <p>Results</p> <p>Deletion of the <it>rcsB</it>, which encoded an activator for the production of capsular polysaccharide, had a minor effect on <it>K. pneumoniae </it>resistance to polymyxin B. On the other hand, deletion of <it>ugd </it>or <it>pmrF </it>gene resulted in a drastic reduction of the resistance. The polymyxin B resistance was shown to be regulated by the two-component response regulators PhoP and PmrA at low magnesium and high iron, respectively. Similar to the control identified in <it>Salmonella</it>, expression of <it>pmrD </it>in <it>K. pneumoniae </it>was dependent on PhoP, the activated PmrD would then bind to PmrA to prolong the phosphorylation state of the PmrA, and eventually turn on the expression of <it>pmr </it>for the resistance to polymyxin B.</p> <p>Conclusions</p> <p>The study reports a role of the capsular polysaccharide level and the <it>pmr </it>genes for <it>K. pneumoniae </it>resistance to polymyxin B. The PmrD connector-mediated pathway in governing the regulation of <it>pmr </it>expression was demonstrated. In comparison to the <it>pmr </it>regulation in <it>Salmonella</it>, PhoP in <it>K. pneumoniae </it>plays a major regulatory role in polymyxin B resistance.</p

Characterization of novel transforming growth factor-beta type I receptors found in malignant pleural effusion tumor cells

<p>Abstract</p> <p>Background</p> <p>Tumors expressing a transforming growth factor-beta type I receptor (TβRI) mutant with sequence deletions in a nine-alanine (9A) stretch of the signal peptide are reported to be highly associated with disease progression. Expression of this mutant could interfere with endogenous TGFβ signaling in the cell. However, little is known about the importance of the remaining part of the signal peptide on the cellular function of TβRI.</p> <p>Results</p> <p>We cloned and identified four new in-frame deletion variants of TβRI, designated DM1 to DM4, in pleural effusion-derived tumor cells. Intriguingly, DM1 and DM2, with a small region truncated in the putative signal peptide of TβRI, had a serious defect in their protein expression compared with that of the wild-type receptor. Using serial deletion mutagenesis, we characterized a region encoded by nucleotides 16–51 as a key element controlling TβRI protein expression. Consistently, both DM1 and DM2 have this peptide deleted. Experiments using cycloheximde and MG132 further confirmed its indispensable role for the protein stability of TβRI. In contrast, truncation of the 9A-stretch itself or a region downstream to the stretch barely affected TβRI expression. However, variants lacking a region C-terminal to the stretch completely lost their capability to conduct TGFβ-induced transcriptional activation. Intriguingly, expression of DM3 in a cell sensitive to TGFβ made it significantly refractory to TGFβ-mediated growth inhibition. The effect of DM3 was to ablate the apoptotic event induced by TGFβ.</p> <p>Conclusion</p> <p>We identified four new transcript variants of TβRI in malignant effusion tumor cells and characterized two key elements controlling its protein stability and transcriptional activation. Expression of one of variants bestowed cancer cells with a growth advantage in the presence of TGFβ. These results highlight the potential roles of some naturally occurring TβRI variants on the promotion of tumor malignancy.</p

Self-supervised learning-based general laboratory progress pretrained model for cardiovascular event detection

The inherent nature of patient data poses several challenges. Prevalent cases amass substantial longitudinal data owing to their patient volume and consistent follow-ups, however, longitudinal laboratory data are renowned for their irregularity, temporality, absenteeism, and sparsity; In contrast, recruitment for rare or specific cases is often constrained due to their limited patient size and episodic observations. This study employed self-supervised learning (SSL) to pretrain a generalized laboratory progress (GLP) model that captures the overall progression of six common laboratory markers in prevalent cardiovascular cases, with the intention of transferring this knowledge to aid in the detection of specific cardiovascular event. GLP implemented a two-stage training approach, leveraging the information embedded within interpolated data and amplify the performance of SSL. After GLP pretraining, it is transferred for TVR detection. The proposed two-stage training improved the performance of pure SSL, and the transferability of GLP exhibited distinctiveness. After GLP processing, the classification exhibited a notable enhancement, with averaged accuracy rising from 0.63 to 0.90. All evaluated metrics demonstrated substantial superiority (p < 0.01) compared to prior GLP processing. Our study effectively engages in translational engineering by transferring patient progression of cardiovascular laboratory parameters from one patient group to another, transcending the limitations of data availability. The transferability of disease progression optimized the strategies of examinations and treatments, and improves patient prognosis while using commonly available laboratory parameters. The potential for expanding this approach to encompass other diseases holds great promise.Comment: published in IEEE Journal of Translational Engineering in Health & Medicin

Rapid Detection of Heterogeneous Vancomycin-Intermediate Staphylococcus aureus Based on Matrix-Assisted Laser Desorption Ionization Time-of-Flight: Using a Machine Learning Approach and Unbiased Validation

Heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) is an emerging superbug with implicit drug resistance to vancomycin. Detecting hVISA can guide the correct administration of antibiotics. However, hVISA cannot be detected in most clinical microbiology laboratories because the required diagnostic tools are either expensive, time consuming, or labor intensive. By contrast, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) is a cost-effective and rapid tool that has potential for providing antibiotics resistance information. To analyze complex MALDI-TOF mass spectra, machine learning (ML) algorithms can be used to generate robust hVISA detection models. In this study, MALDI-TOF mass spectra were obtained from 35 hVISA/vancomycin-intermediate S. aureus (VISA) and 90 vancomycin-susceptible S. aureus isolates. The vancomycin susceptibility of the isolates was determined using an Etest and modified population analysis profile–area under the curve. ML algorithms, namely a decision tree, k-nearest neighbors, random forest, and a support vector machine (SVM), were trained and validated using nested cross-validation to provide unbiased validation results. The area under the curve of the models ranged from 0.67 to 0.79, and the SVM-derived model outperformed those of the other algorithms. The peaks at m/z 1132, 2895, 3176, and 6591 were noted as informative peaks for detecting hVISA/VISA. We demonstrated that hVISA/VISA could be detected by analyzing MALDI-TOF mass spectra using ML. Moreover, the results are particularly robust due to a strict validation method. The ML models in this study can provide rapid and accurate reports regarding hVISA/VISA and thus guide the correct administration of antibiotics in treatment of S. aureus infection

Sarcocrassocolides M–O, Bioactive Cembranoids from the Dongsha Atoll Soft Coral Sarcophyton crassocaule

Three new cembranoids, sarcocrassocolides M–O (1–3), have been isolated from the soft coral Sarcophyton crassocaule. The structures of the metabolites were determined by extensive spectroscopic analysis. Compounds 1–3 were shown to exhibit moderate cytotoxicity toward a limited panel of cancer cell lines and display significant in vitro anti-inflammatory activity in LPS-stimulated RAW264.7 macrophage cells by inhibiting the expression of the iNOS protein

Stripe Helical Magnetism and Two Regimes of Anomalous Hall Effect in NdAlGe

We report the magnetic and electronic transport properties of the inversion and time-reversal symmetry breaking Weyl semimetal NdAlGe. This material is analogous to NdAlSi, whose helical magnetism presents a rare example of a Weyl-mediated collective phenomenon, but with a larger spin-orbit coupling. Our neutron diffraction experiments revealed that NdAlGe, similar to NdAlSi, supports an incommensurate Ising spin density wave ($T_{\text{inc}}=6.8$ K) with a small helical spin canting of 3$^\circ$ and a long-wavelength of $\sim$ 35 nm, which transitions to a commensurate ferrimagnetic state below $T_{\text{com}}=5.1$ K. Using small-angle neutron scattering, we showed that the zero-field cooled ferrimagnetic domains form stripes in real space with characteristic length scales of 18 nm and 72 nm parallel and perpendicular to the [110] direction, respectively. Interestingly, for the transport properties, NdAlSi does not exhibit an anomalous Hall effect (AHE) that is commonly observed in magnetic Weyl semimetals. In contrast to NdAlSi, we identify two different AHE regimes in NdAlGe that are respectively governed by intrinsic Berry curvature and extrinsic disorders/spin fluctuations. Our study suggests that Weyl-mediated magnetism prevails in this group of noncentrosymmetric magnetic Weyl semimetals NdAl$X$, but transport properties including AHE are affected by material-specific extrinsic effects such as disorders, despite the presence of prominent Berry curvature.Comment: Preprint, 16 pages, 6 main figures, 6 supplementary figure