Twenty-one Serratia marcescens, ten Klebsiella pneumoniae, and one Escherichia coli isolate with carbapenem resistance or reduced carbapenem susceptibility were recovered from intensive care units (ICUs) in our hospital. Enterobacterial repetitive intergenic consensus-PCR and pulsed-field gel electrophoresis demonstrated that all the S. marcescens isolates belonged to a clonal strain and the 10 K. pneumoniae isolates were indistinguishable or closely related to each other. The MICs of imipenem, meropenem, and ertapenem for all isolates were 2 to 8 g/ml, except for K. pneumoniae K10 (MICs of 128, 256, and >256 g/ml). Isoelectric focusing, PCRs, and DNA sequencing indicated that all S. marcescens isolates produced KPC-2 and a ␤-lactamase with a pI of 6.5. All K. pneumoniae isolates produced TEM-1, KPC-2, CTX-M-14, and a ␤-lactamase with a pI of 7.3. The E. coli E1 isolate produced KPC-2, CTX-M-15, and a ␤-lactamase with a pI of 7.3. Conjugation studies with E. coli (EC600) resulted in the transfer of reduced carbapenem susceptibility compared to that of the original isolates, and only the bla KPC-2 gene was detected in E. coli transconjugants. Plasmid restriction analysis showed identical restriction patterns among all E. coli transconjugants. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ompK35/36 gene sequence analysis of outer membrane proteins revealed that K. pneumoniae K10 failed to express OmpK36, because of insertional inactivation by an insertion sequence ISEcp1. All these results indicate that KPC-2-producing S. marcescens, K. pneumoniae, and E. coli isolates emerged in ICUs in our hospital. KPC-2 combined with porin deficiency results in high-level carbapenem resistance in K. pneumoniae. The same bla KPC-2 -encoding plasmid was spread among the three different genera.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is emerging as a worldwide public health concern; however, the longterm molecular epidemiological surveillance of clinical CRKP in China is limited. We conducted a retrospective observational study (2008-2018) to assess the prevalence, susceptibility, risk factors and molecular epidemiology of clinical CRKP isolates. We found the prevalence of CRKP increased from 2.5%, 2008 to 15.8%, 2018. CRKP were significantly more frequent among hospitalized patients from ICU, and it was significantly more likely to be isolated from the capital city (Hangzhou) and the patients aged ≥60 years. Additionally, seasons and specimen types were associated with CRKP infections. The main CRKP sequence type (ST) was ST11, and bla KPC-2 was the most prevalent gene variant. Together these data reveal an increasing incidence and resistance trends among CRKP, especially the ST11-bla KPC-2-CRKP, in Zhejiang, during 2008-2018. Our findings are important for hospitals to limit its dissemination and optimize antibiotic administration.

Simulation systems have become an essential component in the development and validation of autonomous driving technologies. The prevailing state-of-the-art approach for simulation is to use game engines or high-fidelity computer graphics (CG) models to create driving scenarios. However, creating CG models and vehicle movements (a.k.a. the assets for simulation) remains a manual task that can be costly and time-consuming. In addition, the fidelity of CG images still lacks the richness and authenticity of real-world images and using these CG images for training leads to degraded performance.In this paper we present a novel approach to address these issues: Augmented Autonomous Driving Simulation (AADS). Our formulation augments real-world pictures with a simulated traffic flow to create photo-realistic simulation images and renderings. More specifically, we use LiDAR and cameras to scan street scenes. From the acquired trajectory data, we generate highly plausible traffic flows for cars and pedestrians and compose them into the background. The composite images can be re-synthesized with different viewpoints and sensor models (camera or LiDAR). The resulting images are photo-realistic, fully annotated, and ready for end-to-end training and testing of autonomous driving systems from perception to planning. We explain our system design and validate our algorithms with a number of autonomous driving tasks from detection to segmentation and predictions.Compared to traditional approaches, our method offers unmatched scalability and realism. Scalability is particularly important for AD simulation and we believe the complexity and diversity of the real world cannot be realistically captured in a virtual environment. Our augmented approach combines the flexibility of a virtual environment (e.g., vehicle movements) with the richness of the real world to allow effective simulation of any location on earth. Summary

Klebsiella pneumoniae has emerged as one of the most important pathogens that frequently encounter in community-acquired or hospital-acquired infections. Timely epidemiological surveillance could greatly facilitate infection control of K. pneumoniae and many deadly pathogens alike. In this study, we evaluated the performance of the IR Biotyper, a Fourier transform infrared (FTIR) spectroscopy system for K. pneumoniae isolates typing through (i) optimizing the culture scheme and defining the cutoff value (COV) range and (ii) comparing with commonly used typing tools such as multi-locus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and whole-genome sequencing (WGS). We found that a non-selective and non-chromogenic medium with 24 AE 2 h incubation gives the best discriminatory power for the IR Biotyper (IRBT). COV evaluation indicated that the IRBT is a robust typing method with good reproducibility. Besides, we observed that the modified H 2 O-EtOH suspensions preparation method could enhance the quality of the spectrum, especially for those hypermucoviscous strains. For the method comparison study, our data demonstrated that FTIR spectroscopy could accurately cluster K. pneumoniae strains. The typing results of the IRBT were almost entirely in concordance with those from PFGE and WGS. Together with the advantages such as low costs and short turnaround time (less than 3h), the IRBT is a promising tool for strain typing that could make real-time outbreak investigation a reality.

Two carbapenem-non-susceptible Klebsiella pneumoniae isolates, Z2554 and Z2110, were collected from a hospital in China and analysed by PFGE. K. pneumoniae Z2554 and Z2110 were genetically unrelated and showed resistance to ertapenem, and reduced susceptibility to imipenem and meropenem. Analysis of their b-lactamases indicated that K. pneumoniae Z2554 produced TEM-1 and CTX-M-14 b-lactamases, whilst Z2110 produced a plasmid-mediated AmpC b-lactamase, DHA-1, in addition to TEM-1 and CTX-M-14. SDS-PAGE analysis of the outer-membrane proteins (OMPs) revealed that both isolates lacked an OMP of~39 kDa (OmpK36), whilst Z2110 had an additional protein with an approximate molecular mass of 26 kDa. Analysis of the OMP-encoding genes demonstrated that the ompK35 sequence of K. pneumoniae Z2554 and Z2110 contained a number of silent mutations. In ompK36, several insertions and deletions of short DNA fragments (1-6 bp) were detected in both isolates. The Nterminal sequence of the~26 kDa protein band identified in Z2110 had no similarity to the sequence of OmpK36. Instead, it shared high similarity with hypothetical protein KPN_03267 originating from K. pneumoniae subsp. pneumoniae MGH 78578. It was concluded that blactamase production combined with OmpK36 deficiency results in ertapenem resistance, and reduced imipenem and meropenem susceptibility, in K. pneumoniae Z2554 and Z2110. OmpK36 may play an important role in the resistance or reduced susceptibility to carbapenems in K. pneumoniae producing AmpC, extended-spectrum b-lactamase or broad-spectrum b-lactamase.