Genomic characterization of the novel bacteriophage IME183, infecting Klebsiella pneumoniae of capsular type K2.

Klebsiella pneumoniae causes a wide range of serious and life-threatening infections. Klebsiella phage IME183, isolated from hospital sewage, exhibited lytic activity against K. pneumoniae of capsular type K2. Transmission electron microscopy revealed that phage IME183 has a head with a diameter of 50 nm and a short tail. Its genome is 41,384 bp in length with a GC content of 52.92%. It is predicted to contain 50 open reading frames (ORFs). The results of evolutionary analysis suggest that phage IME183 should be considered a member of a new species in the genus Przondovirus.

Analysis of the diversity of tick-borne viruses at the border areas in Liaoning Province, China.

Ticks play a significant role in transmitting arboviruses, which pose a risk to human and animal health. The region of Liaoning Province, China, with abundant plant resources with multiple tick populations, has reported several tick-borne diseases. However, there remains a scarcity of research on the composition and evolution of the tick virome. In this study, we conducted the metagenomic analysis of 561 ticks in the border area of Liaoning Province in China and identified viruses related to known diseases in humans and animals, including severe fever with thrombocytopenia syndrome virus (SFTSV) and nairobi sheep disease virus (NSDV). Moreover, the groups of tick viruses were also closely related to the families of Flaviviridae, Parvoviridae, Phenuiviridae, and Rhabdoviridae. Notably, the Dabieshan tick virus (DBTV) of the family Phenuiviridae was prevalent in these ticks, with the minimum infection rate (MIR) of 9.09%, higher than previously reported in numerous provinces in China. In addition, sequences of tick-borne viruses of the family Rhabdoviridae have first been reported from the border area of Liaoning Province, China, after being described from Hubei Province, China. This research furthered the insight into pathogens carried by ticks in the northeastern border areas of China, offering epidemiological information for possible forthcoming outbreaks of infectious diseases. Meanwhile, we provided an essential reference for assessing the risk of tick bite infection in humans and animals, as well as for exploring into the evolution of the virus and the mechanisms of species transmission.

Genomic representation predicts an asymptotic host adaptation of bat coronaviruses using deep learning.

Introduction: Coronaviruses (CoVs) are naturally found in bats and can occasionally cause infection and transmission in humans and other mammals. Our study aimed to build a deep learning (DL) method to predict the adaptation of bat CoVs to other mammals. Methods: The CoV genome was represented with a method of dinucleotide composition representation (DCR) for the two main viral genes, ORF1ab and Spike. DCR features were first analyzed for their distribution among adaptive hosts and then trained with a DL classifier of convolutional neural networks (CNN) to predict the adaptation of bat CoVs. Results and discussion: The results demonstrated inter-host separation and intra-host clustering of DCR-represented CoVs for six host types: Artiodactyla, Carnivora, Chiroptera, Primates, Rodentia/Lagomorpha, and Suiformes. The DCR-based CNN with five host labels (without Chiroptera) predicted a dominant adaptation of bat CoVs to Artiodactyla hosts, then to Carnivora and Rodentia/Lagomorpha mammals, and later to primates. Moreover, a linear asymptotic adaptation of all CoVs (except Suiformes) from Artiodactyla to Carnivora and Rodentia/Lagomorpha and then to Primates indicates an asymptotic bats-other mammals-human adaptation. Conclusion: Genomic dinucleotides represented as DCR indicate a host-specific separation, and clustering predicts a linear asymptotic adaptation shift of bat CoVs from other mammals to humans via deep learning.

Survey of mosquito species and mosquito-borne viruses in residential areas along the Sino-Vietnam border in Yunnan Province in China.

Mosquitoes are capable of carrying complex pathogens, and their feeding habits on the mammalian blood can easily mediate the spread of viruses. Surveillance of mosquito-based arbovirus enables the early prevention and control of mosquito-borne arboviral diseases. The climate and geography of Yunnan Province in China are ideal for mosquitoes. Yunnan shares borders with several other countries; therefore, there exists a high risk of international transmission of mosquito-mediated infectious diseases. Previous studies have focused more on the Sino-Laos and Sino-Myanmar borders. Therefore, we focused on the neighborhoods of Malipo and Funing counties in Wenshan Prefecture, Yunnan Province, China, which are located along the Sino-Vietnam border, to investigate the species of mosquitoes and mosquito-borne viruses in the residential areas of this region. This study collected 10,800 mosquitoes from 29 species of 8 genera and grouped to isolate mosquito-borne viruses. In total, 62 isolates were isolated and classified into 11 viral categories. We demonstrated a new distribution of mosquito-borne viruses among mosquitoes in border areas, including Tembusu and Getah viruses, which can cause animal outbreaks. In addition, Dak Nong and Sarawak viruses originating from Vietnam and Malaysia, respectively, were identified for the first time in China, highlighting the complexity of mosquito-borne viruses in the Sino-Vietnam border region. The awareness of the importance of viral surveillance and prevention measures in border areas should be further encouraged to prevent future outbreaks of potentially infectious diseases.

The diagnosis and treatment for a patient with cancer of unknown primary: A case report.

Background: Cancer of unknown primary (CUP) is a class of metastatic malignant tumors whose primary location cannot be determined. The diagnosis and treatment of CUP are a considerable challenge for clinicians. Herein, we report a CUP case whose corresponding primary tumor sites were successfully identified, and the patient received proper treatment. Case report: In February 2022, a 74-year-old woman was admitted to the Medical Oncology Department at Sir Run Run Shaw Hospital for new lung and intestinal tumors after more than 9 years of breast cancer surgery. After laparoscopically assisted right hemicolectomy, pathology revealed mucinous adenocarcinoma; the pathological stage was pT2N0M0. Results from needle biopsies of lung masses suggested poorly differentiated cancer, ER (-), PR (-), and HER2 (-), which combined with the clinical history, did not rule out metastatic breast cancer. A surgical pathology sample was needed to determine the origin of the tumor tissue, but the patient's chest structure showed no indications for surgery. Analysis of the tumor's traceable gene expression profile prompted breast cancer, and analysis of next-generation amplification sequencing (NGS) did not obtain a potential drug target. We developed a treatment plan based on comprehensive immunohistochemistry, a gene expression profile, and NGS analysis. The treatment plan was formulated using paclitaxel albumin and capecitabine in combination with radiotherapy. The efficacy evaluation was the partial response (PR) after four cycles of chemotherapy and two cycles combined with radiotherapy. Conclusion: This case highlighted the importance of identifying accurate primary tumor location for patients to benefit from treatment, which will provide a reference for the treatment decisions of CUP tumors in the future.

Characterization of the novel temperate Staphylococcus haemolyticus phage IME1365_01.

The presence of a novel functional prophage, IME1365_01, was predicted from bacterial high-throughput sequencing data and then successfully induced from Staphylococcus haemolyticus by mitomycin C treatment. Transmission electron microscopy showed that phage IME1365_01 has an icosahedral head (43 nm in diameter) and a long tail (172 nm long). This phage possesses a double-stranded DNA genome of 44,875 bp with a G+C content of 35.35%. A total of 63 putative open reading frames (ORFs) were identified in its genome. BLASTn analysis revealed that IME1365_01 is similar to Staphylococcus phage vB_SepS_E72, but with a genome homology coverage of only 26%. The phage genome does not have fixed termini. In ORF24 of phage IME1365_01, a conserved Toll-interleukin-1 receptor domain of the TIR_2 superfamily (accession no. c123749) is located at its N-terminus, and this might serve as a component of an anti-bacterial system. In conclusion, we developed a platform to obtain active temperate phage from prediction, identification, and induction from its bacterial host. After mass screening using this platform, numerous temperate phages and their innate anti-bacterial elements can provide extensive opportunities for therapy against bacterial (especially drug-resistant bacterial) infections.

A SARS-CoV-2-Related Virus from Malayan Pangolin Causes Lung Infection without Severe Disease in Human ACE2-Transgenic Mice.

Coronavirus disease 2019 (COVID-19), which is caused by the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is the most severe emerging infectious disease in the current century. The discovery of SARS-CoV-2-related coronaviruses (SARSr-CoV-2) in bats and pangolins in South Asian countries indicates that SARS-CoV-2 likely originated from wildlife. To date, two SARSr-CoV-2 strains have been isolated from pangolins seized in Guangxi and Guangdong by the customs agency of China, respectively. However, it remains unclear whether these viruses cause disease in animal models and whether they pose a transmission risk to humans. In this study, we investigated the biological features of a SARSr-CoV-2 strain isolated from a smuggled Malayan pangolin (Manis javanica) captured by the Guangxi customs agency, termed MpCoV-GX, in terms of receptor usage, cell tropism, and pathogenicity in wild-type BALB/c mice, human angiotensin-converting enzyme 2 (ACE2)-transgenic mice, and human ACE2 knock-in mice. We found that MpCoV-GX can utilize ACE2 from humans, pangolins, civets, bats, pigs, and mice for cell entry and infect cell lines derived from humans, monkeys, bats, minks, and pigs. The virus could infect three mouse models but showed limited pathogenicity, with mild peribronchial and perivascular inflammatory cell infiltration observed in lungs. Our results suggest that this SARSr-CoV-2 virus from pangolins has the potential for interspecies infection, but its pathogenicity is mild in mice. Future surveillance among these wildlife hosts of SARSr-CoV-2 is needed to monitor variants that may have higher pathogenicity and higher spillover risk. IMPORTANCE SARS-CoV-2, which likely spilled over from wildlife, is the third highly pathogenic human coronavirus. Being highly transmissible, it is perpetuating a pandemic and continuously posing a severe threat to global public health. Several SARS-CoV-2-related coronaviruses (SARSr-CoV-2) in bats and pangolins have been identified since the SARS-CoV-2 outbreak. It is therefore important to assess their potential of crossing species barriers for better understanding of their risk of future emergence. In this work, we investigated the biological features and pathogenicity of a SARSr-CoV-2 strain isolated from a smuggled Malayan pangolin, named MpCoV-GX. We found that MpCoV-GX can utilize ACE2 from 7 species for cell entry and infect cell lines derived from a variety of mammalian species. MpCoV-GX can infect mice expressing human ACE2 without causing severe disease. These findings suggest the potential of cross-species transmission of MpCoV-GX, and highlight the need of further surveillance of SARSr-CoV-2 in pangolins and other potential animal hosts.

Virome in healthy pangolins reveals compatibility with multiple potentially zoonotic viruses.

Previous studies have identified multiple viruses in dead or severely diseased pangolins, but descriptions of the virome in healthy pangolins are lacking. This poses a greater risk of cross-species transmission due to poor preventive awareness and frequent interactions with breeders. In this study, we investigated the viral composition of 34 pangolins with no signs of disease at the time of sampling and characterized a large number of arthropod-associated viruses belonging to 11 families and vertebrate viruses belonging to eight families, including those with pathogenic potential in humans and animals. Several important vertebrate viruses were identified in the pangolins, including parvovirus, pestivirus, and picobirnavirus. The picobirnavirus was clustered with human and grey teal picobirnaviruses. Viruses with cross-species transmission ability were also identified, including circovirus, rotavirus, and astrovirus. Our study revealed that pangolins are frequently exposed to arthropod-associated viruses in the wild and can carry many vertebrate viruses under natural conditions. This study provides important insights into the virome of pangolins, underscoring the importance of monitoring potential pathogens in healthy pangolins to prevent outbreaks of infectious diseases in domesticated animals and humans.

N-glycoproteomic profiling revealing novel coronavirus therapeutic targets potentially involved in Cepharanthine's intervention.

The Coronavirus disease 2019 (COVID-19) has posed a serious threat to global health and the world economy. Antiviral therapies targeting coronavirus are urgently required. The Cepharanthine (CEP) is a traditional Chinese herbal extract. Our previous research revealed that CEP has a very potent anti-coronavirus effect, but its mechanism of action was not fully understood. To investigate the effect of novel coronavirus on protein glycosylation in infected cells and to further investigate the mechanism of action of CEP against coronavirus, a cellular model using coronavirus GX_P2V infection of Vero E6 cells was established. The effect of coronavirus GX_P2V on host cell protein glycosylation was investigated by N-glycoproteomic analysis, and the antagonistic effect of CEP on the abnormal protein glycosylation caused by coronavirus was analyzed. The results showed that GX_P2V could cause abnormal changes in protein glycosylation levels in host cells, while CEP could partially antagonize the abnormal protein glycosylation caused by GX_P2V. In addition, we also found that CEP could regulate the glycosylation level of coronavirus S protein. In conclusion, this article provides important ideas about the infection mechanism of novel coronaviruses, providing evidence for CEP as a promising therapeutic option for coronavirus infection.

Isolation and genomic characterization of a novel Autographiviridae bacteriophage IME184 with lytic activity against Klebsiella pneumoniae.

Klebsiella pneumoniae, a multidrug resistant bacterium that causes nosocomial infections including septicemia, pneumonia etc. Bacteriophages are potential antimicrobial agents for the treatment of antibiotic resistant bacteria. In this study, a novel bacteriophage IME184, was isolated from hospital sewage against clinical multi-drug resistant Klebsiella pneumoniae. Transmission electron microscopy and genomic characterization exhibited this phage belongs to the Molineuxvirinae genus, Autographiviridae family. Phage IME184 possessed a double-stranded DNA genome composed of 44,598 bp with a GC content of 50.3%. The phage genome encodes 57 open reading frames, out of 26 are hypothetical proteins while 31 had assigned putative functions. No tRNA, virulence related or antibiotic resistance genes were found in phage genome. Comparative genomic analysis showed that phage IME184 has 94% similarity with genomic sequence of Klebsiella phage K1-ULIP33 (MK380014.1). Multiplicity of infection, one step growth curve and host range of phage were also measured. According to findings, Phage IME184 is a promising biological agent that infects Klebsiella pneumoniae and can be used in future phage therapies.

Molecular dissection of the first Staphylococcus cohnii temperate phage IME1354_01.

In staphylococcal phage research, studies specific to coagulase-negative staphylococci (CoNS) remain severely under-represented, and the number of temperate bacteriophages is limited. This investigation identifies a novel temperate phage IME1354_01 from the strain Staphylococcus cohnii IME1354, which was isolated from the skin of a patient with foot ulcer disease. The phage IME1354_01 is the first isolated temperate phage of S. cohnii, and was determined to have a long-tail morphology using TEM. Its genome was found to be a 42,706-bp linear dsDNA molecule with a GC content of 34%. The integration of IME1354_01 occurred using a tRNA-Ser coding gene, and it did not affect tRNA-Ser function. The genome of IME1354_01 is most closely related to that of the temperate Staphylococcus arlettae phage vB_Sars_BM31 with 10% homology coverage and 83.73% nucleotide identity. In addition, they showed similarities mainly in the DNA replication, DNA packaging and partial morphogenesis modules. We propose that a new genus should be created for IME1354_01 based on the intergenomic similarities (maximum is 23%) obtained from the VIRIDIC calculations. The isolation and in-depth study of the novel phage, IME1354_01, will improve our understanding of the evolutionary relationship between temperate phages and their hosts.

Characterization and complete genome sequence analysis of a newly isolatedphage against Vibrio parahaemolyticus from sick shrimp in Qingdao, China.

Foodborne diseases have become a serious havoc, where antimicrobial resistance is throwing significant challenges on daily basis. With the increase of drug-resistant bacteria and food-borne infection associated with Vibrio parahaemolyticus, new and effective strategies were needed to control the emergence of vibriosis. Lytic bacteriophages come up as a promising way to resist the pathogenic population in various applications. In this study, a V. parahaemolyticus specific phage vB_VpS_PG28 was isolated from sewage in the seafood market. Results showed vB_VpS_PG28, is strictly a lytic bacteriophage and has a relatively large burst size of 103 plaque-forming units per infected cell. Comparative genomic and bioinformatic analyses proved that vB_VpS_PG28 is a new bacteriophage that had a homologous relation with Vibrio phages of family Siphoviridae, especially with phage VH2_2019, but transmission electron microscopy of vB_VpS_PG28 morphology characterized its morphology is similar to that of Myoviridae family. In silico analysis indicated that the vB_VpS_PG28 genome consists of 82712 bp (48.08% GC content) encoding 114 putative ORFs without tRNA,and any gene associated with resistance or virulence factors has not been found. The bacteriophage in the present study has shown significant outcomes in order to control bacterial growth under in vitro conditions. Thus, we are suggesting a beneficiary agent against foodborne pathogens. Further, to ensure the safe usage of phage oral toxicity testing is recommended.

Evaluation of Modified Calcium Removal Algorithm in dual energy CT of Internal Carotid Artery.

PURPOSE: To evaluate the performance of a dual-energy (DE) calcium removal software based on a modified three-material decomposition algorithm in assessing the stenosis of the internal carotid artery (ICA) in comparison with mixed images using digital subtraction angiography (DSA) as the reference standard. METHODS: Forty-six patients (38 men; 67±8 years old), including 154 calcified ICA segments C1-C2 (59), C3-C5 (63), C6 (24), and C7 (8), were recruited in this retrospective study. Mixed images and virtual non-calcium (VNCa) images using the modified dual-energy computed tomography (DECT) algorithm were reconstructed. The differences between VNCa and DSA images vs. mixed and DSA images of degree of stenosis were compared. The intraclass correlation coefficient (ICC) was used for assessing the agreement between VNCa, mixed images, and DSA. RESULTS: The degree of stenosis differed significantly between mixed and DSA images in the C3-C5 (30%±17.9% vs. 23.0%±16.9%, p = 0.026) and C6 (38.3%±15.4% vs. 28.5%±13.3%, p = 0.023) segments. The stenosis of VNCa images showed no significant difference with DSA images in all segments (all p > 0.05). The ICCs between VNCa and DSA images (0.86-0.97) were higher than those between the mixed and DSA images (0.68-0.96) in all segments. CONCLUSION: The performance of a modified three-material decomposition DECT algorithm for calcium removal in ICA stenosis evaluation, particularly for the C3-C5 and C6 ICA segments, was promising.

Characteristics and genome analysis of a novel bacteriophage IME1323_01, the first temperate bacteriophage induced from Staphylococcus caprae.

Temperate phages play an important role in the evolution of bacteria. So far, lytic phages have been wildly reported, but there is still limited knowledge regarding temperate phages in the genome of pathogenic Staphylococcus caprae. Here we present the characteristics and genome analysis of a novel bacteriophage IME1323_01, which is the first isolated bacteriophage of S. caprae. The phage genome is a 44282-bp linear dsDNA molecule with a GC content of 34.18%, which is similar to its host. The genome of IME1323_01 is most closely related with that of temperate phage IME1318_01, whereas the homology coverage is just 34%. Genome and proteome analyses confirmed the lysogenic nature of phage IME1323_01, which encodes the typical lysogen-related proteins integrase, CI, Cro, and anti-repressor proteins. Genomic and phylogenetic analysis revealed that phage IME1323_01 is a newly discovered phage, which belongs to subfamily Azeredovirinae in the family Siphoviridae. The goal of this study is to increase our knowledge about the phages of S. caprae and expand our armamentarium against the escalating threat of pathogenic bacteria.

First report of norovirus sequences isolated from raccoon dogs in mainland China.

Noroviruses can infect humans and a wide variety of other mammalian hosts, causing varying degrees of diarrhea. In this study, two novel norovirus genomes were identified for the first time in farmed raccoon dogs, designated as raccoon dog noroviruses BUCT-K1 and BUCT-K4. Neither the farmers nor the raccoon dogs had symptoms (e.g., diarrhea) at the time of sample collection. We collected 14 stool samples from two farms, and 85.7% (12/14) of the samples were norovirus positive by RT-PCR. The two norovirus genomes have the highest identity to Dog/Z7/19/CH, suggesting that the norovirus might have been transmitted from dogs to raccoon dogs. Genomic and evolutionary analyses indicated that different directions of evolution occurred following the spread of the norovirus to the raccoon dogs. This study has increased knowledge of norovirus-infected animal species and has provided additional information on the norovirus family.

Characterization and Genomic Analysis of BUCT549, a Novel Bacteriophage Infecting Vibrio alginolyticus With Flagella as Receptor.

Vibrio alginolyticus is one of the most important of pathogens that can infect humans and a variety of aquatic animals, and it can cause food poisoning and septicemia in humans. Widely used antibiotics are gradually losing their usefulness, and phages are gaining more attention as new antibacterial strategies. To have more potential strategies for controlling pathogenic bacteria, we isolated a novel V. alginolyticus phage BUCT549 from seafood market sewage. It was classified as a new member of the family Siphoviridae by transmission electron microscopy and a phylogenetic tree. We propose creating a new genus for BUCT549 based on the intergenomic similarities (maximum is 56%) obtained from VIRIDIC calculations. Phage BUCT549 could be used for phage therapy due to its stability in a wide pH (3.0-11.0) range and high-temperature (up to 60°C) environment. It had a latent period of 30-40 min and a burst size of 141 PFU/infected bacterium. In the phylogenetic tree based on a terminase large subunit, BUCT549 was closely related to eight Vibrio phages with different species of host. Meanwhile, our experiments proved that BUCT549 has the ability to infect a strain of Vibrio parahaemolyticus. A coevolution experiment determined that three strains of tolerant V. alginolyticus evaded phage infestation by mutating the MSHA-related membrane protein expression genes, which caused the loss of flagellum. This research on novel phage identification and the mechanism of infestation will help phages to become an integral part of the strategy for biological control agents.

A novel CT-based radiomic nomogram for predicting the recurrence and metastasis of gastric stromal tumors.

Our study aimed to explore the value of applying the CT-based radiomic nomogram for predicting recurrence and/or metastasis (RM) of gastric stromal tumors (GSTs). During the past ten years, a total of 236 patients with GST were analyzed retrospectively. According to the postoperative follow-up classification, the patients were divided into two groups, namely non-recurrence/metastasis group (non-RM) and RM group. All the cases were randomly divided into primary cohort and validation cohort according to the ratio of 7:3. Standardized CT images were segmented by radiologists using ITK-SNAP software manually. Texture features were extracted from all segmented lesions, then radiomic features were selected and the radiomic nomogram was built using least absolute shrinkage and selection operator (LASSO) method. The clinical features with the greatest correlation with RM of GST were selected by univariate analysis, and used as parameters to build the clinical feature model. Eventually, model of radiomic and clinical features were fitted to construct the clinical + radiomic feature model. The performance of each model was evaluated by the area under receiver operating characteristic (ROC) curve (AUC). A total of 1223 features were extracted from all the segmentation regions of each case, and features were selected via the least absolute shrinkage and LASSO binary logistic regression model. After deletion of redundant features, four key features were obtained, which were used as the parameters to build a radiomic signature. The AUCs of radiomic nomogram in primary cohort and validation cohort were 0.816 and 0.946, respectively. The AUCs of clinical + radiomic feature model in primary cohort and validation cohort were 0.833 and 0.937, respectively. Using DeLong test, the differences of AUC values between radiomic nomogram and clinical + radiomic feature model in primary cohort (P = 0.840) and validation cohort (P = 0.857) were not statistically significant. To sum up, CT-based radiomic nomogram is of great potential in predicting the RM of GST non-invasively before operation.