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Accurate identification of fungal species is crucial for mycological research, relying significantly on experienced taxonomists’ ability to recognize fungal morphology. With the dwindling number of taxonomists, developing a rapid, accurate, and automated fungal identification method is essential, promising to markedly decrease the time and resources needed for such tasks. To address the challenges in the automatic identification of nematode-trapping fungi (NTF, belong to Orbiliaceae, Orbiliomycetes, a group of filamentous fungi that catch nematodes in soil), specifically focusing on their conidia which are often aggregated and morphologically similar, we developed an enhanced method. Our approach integrates an efficient channel attention (ECA) mechanism within the backbone layer of YOLOv8l, improving the algorithm’s capability to detect global information effectively. This adjustment significantly optimizes the detection and differentiation of limited features and small-sized conidia among aggregated samples. The experimental outcomes demonstrated that our method increases precision by 0.3%, recall by 1.8%, and mean average precision (mAP) by 0.4%, surpassing the performance of existing target detection algorithms. In addition, this method accurately identified the aggregated fungal conidia and distinguished morphologically similar spores. In conclusion, the system accurately detects and recognizes the conidia of NTF in mixed spore environments, thus providing crucial technical support for the automatic detection and early prediction of NTF.

Long COVID, also known for post-acute sequelae of COVID-19, describes the people who have the signs and symptoms that continue or develop after the acute COVID-19 phase. Long COVID patients suffer from an inflammation or host responses towards the virus approximately 4 weeks after initial infection with the SARS CoV-2 virus and continue for an uncharacterized duration. Anyone infected with COVID-19 before could experience long-COVID conditions, including the patients who were infected with SARS CoV-2 virus confirmed by tests and those who never knew they had an infection early. People with long COVID may experience health problems from different types and combinations of symptoms over time, such as fatigue, dyspnea, cognitive impairments, and gastrointestinal (GI) symptoms (e.g., nausea, vomiting, diarrhea, decreased or loss of appetite, abdominal pain, and dysgeusia). The critical role of the microbiome in these GI symptoms and long COVID were reported in clinical patients and experimental models. Here, we provide an overall view of the critical role of the GI tract and microbiome in the development of long COVID, including the clinical GI symptoms in patients, dysbiosis, viral–microbiome interactions, barrier function, and inflammatory bowel disease patients with long COVID. We highlight the potential mechanisms and possible treatment based on GI health and microbiome. Finally, we discuss challenges and future direction in the long COVID clinic and research.

Carbothermal reduction using biochar (BC) is a green and effective method of synthesizing BC-supported nanoscale zero-valent iron (nanoFe0) composites. However, the effect of BC surface area on the structure, distribution, and performance such as the heavy metal uptake capacity of nanoFe0 particles remains unclear. Soybean stover-based BCs with different surface areas (1.7 − 1 472 m2/g) were prepared in this study. They have been used for in-situ synthesis BCs-supported nanoFe0 particles through carbothermal reduction of ferrous chloride. The BCs-supported nanoFe0 particles were found to be covered with graphene shells and dispersed onto BC surfaces, forming the BC-supported graphene-encapsulated nanoFe0 (BC-G@Fe0) composite. These graphene shells covering the nanoFe0 particles were formed because of gaseous carbon evolved from biomass carbonization reacting with iron oxides/iron salts. Increasing BC surface area decreased the average diameters of nanoFe0 particles, indicating a higher BC surface area alleviated the aggregation of nanoFe0 particles, which resulted in higher heavy metal uptake capacity. At the optimized condition, BC-G@Fe0 composite exhibited uptake capacities of 124.4, 121.8, 254.5, and 48.0 mg/g for Cu2+, Pb2+, Ag+, and As3+, respectively (pH 5, 25 °C). Moreover, the BC-G@Fe0 composite also demonstrated high stability for Cu2+ removal from the fixed-bed continuous flow, in which 1 g of BC-G@Fe0 can work for 120 h in a 4 mg/L Cu2+ flow continually and clean 28.6 L Cu2+ contaminated water. Furthermore, the BC-G@Fe0 composite can effectively immobilize the bioavailable As3+ from the contaminated soil, i.e., 5% (w) of BC-G@Fe0 composite addition can immobilize up to 92.2% bioavailable As3+ from the contaminated soil.

High-entropy alloys (HEAs) have attracted considerable attention for their exceptional microstructures and properties. Discovering new HEAs with desirable properties is crucial, but traditional design methods are laborious and time-consuming. Fortunately, the emerging Machine Learning (ML) offers an efficient solution. In this study, composition-microhardness data pairs from various alloy systems were collected and expanded using a Generative Adversarial Network (GAN). These data pairs were converted into empirical parameter-microhardness pairs. Then Active Learning (AL) was employed to screen the Al-Co-Cr-Cu-Fe-Ni system and identify the eXtreme Gradient Boosting (XGBoost) as the optimal ML master model. Millions of data training iterations employing the XGBoost sub-model and accuracy evaluations using the Expected Improvement (EI) algorithm establish the relationship between HEA compositions and microhardness. The proposed sub-model aligns well with experimental data, wherein four Al-rich compositions exhibit ultra-high microhardness (>740 HV, with a maximum of ∼780.3 HV) and low density (

Elastic materials for seat foundations come in a variety of materials, shapes, and dimensions. However, it is difficult to measure the stress-strain relationships of many elastic material combinations by conventional uniaxial compression tests. This study investigated the stress-strain relations of elastic material combinations for foam foundations using finite element analysis (FEA). First, the stress-strain relations of single-layer polymer foams and a combination of double-layer polymer foams with covering were quantified by uniaxial compression tests, and axial tensile tests quantified the properties of the covering material of the fabric. Then, based on the Ogden foam and Ogden constitutive equation of Ansys Workbench 19.2, the test data of single-layer polymer foams and covering were fitted by a non-linear least square method, and a combination of double-layer polymer foams with the covering is predicted by FEA. When the strain was 10% to 65%, the stress error between FEA and test results dropped from 95.68% to -5.08%.

The present study was conducted to determine the effects of dietary phosphorus (P) levels on growth performance, body composition, liver histology and enzymatic activity, and expression of lipid metabolism-related genes in spotted seabass (Lateolabrax maculatus). Seven diets were prepared to contain available P levels of 0.48% (the control group), 0.69%, 0.89%, 1.10%, 1.28%, 1.51% and 1.77% and feed fish (4.26 ± 0.03 g) to satiety twice daily for 10 weeks. Significantly higher weight gain and specific growth rate were recorded at P levels of 0.69%–1.51% compared to the control group. Feed conversion ratio decreased with increasing P levels up to 0.89% and increased thereafter. The lowest liver lipid content, viscerosomatic index and lipid content of whole-body were obtained in the 0.89%-P group among dietary treatments. P and calcium (Ca) contents in whole body were increased, while liver triglyceride and cholesterol contents were decreased with increasing dietary P levels from 0.48% to 1.77%. The highest activity of hepatic lipase was recorded in the 1.10%-P group among dietary treatments. Compared to the control group, 1.10% P enhanced the proportion of HUFA and reduced the proportion of SFA and MUFA. The histological observations showed that P deficiency (0.48%) led to the vacuolization of hepatocytes and increased number of lipid droplets. Meanwhile, overall liver tissue structure was improved when P level increased to 1.28%. Compared to the control group, expression of lipid metabolism-related genes such as FAS, ACC-2 and SREBP-1 was decreased at 0.89%–1.10% P group while an opposite trend was observed in the expression of PPARa2 and CPT-1 genes. The current study showed that 0.89% dietary P levels could promote growth performance of spotted seabass and reduce lipid accumulation in the liver. A broken-line regression analysis based on weight gain showed that the optimum dietary P level (available P) for juvenile spotted seabass reared in freshwater was 0.72%.

Abstract Background Venetoclax (Ven) combined with intensive chemotherapy was proven effective in the management of acute myeloid leukemia (AML). However, the severe and prolonged myelosuppression remains a concern to worry about. To explore more appropriate combination regimens, we designed Ven combining daunorubicin and cytarabine (DA 2 + 6) regimen as induction therapy, aimed to evaluate the effectiveness and safety in adults de novo AML. Methods A phase 2 clinical trial was performed in 10 Chinese hospitals to investigate Ven combined with daunorubicin and cytarabine (DA 2 + 6) in patients with AML. The primary endpoints were overall response rate (ORR), comprising of complete remission (CR), complete remission with incomplete blood cell count recovery (CRi), and partial response (PR). Secondary endpoints included measurable residual disease (MRD) of bone marrow assessed by flow cytometry, overall survival (OS), event-free survival (EFS), disease-free survival (DFS), and the safety of regimens. This study is a currently ongoing trial listed on the Chinese Clinical Trial Registry as ChiCTR2200061524. Results Overall, 42 patients were enrolled from January 2022 to November 2022; 54.8% (23/42) were male, and the median age was 40 (range, 16–60) years. The ORR after one cycle of induction was 92.9% (95% confidence interval [CI], 91.6–94.1; 39/42) with a composite complete response rate (CR + CRi) 90.5% (95% CI, 89.3–91.6, CR 37/42, CRi 1/42). Moreover, 87.9% (29/33) of the CR patients with undetectable MRD (95% CI, 84.9–90.8). Grade 3 or worse adverse effects included neutropenia (100%), thrombocytopenia (100%), febrile neutropenia (90.5%), and one mortality. The median neutrophil and platelet recovery times were 13 (5–26) and 12 (8–26) days, respectively. Until Jan 30, 2023, the estimated 12-month OS, EFS, and DFS rates were 83.1% (95% CI, 78.8–87.4), 82.7% (95% CI, 79.4–86.1), and 92.0% (95% CI, 89.8–94.3), respectively. Conclusion Ven with DA (2 + 6) is a highly effective and safe induction therapy for adults with newly diagnosed AML. To the best of our knowledge, this induction therapy has the shortest myelosuppressive period but has similar efficacy to previous studies.

ABSTRACTThe microbiota plays critical roles in regulating the function and health of the intestine and extraintestinal organs. A fundamental question is whether an intestinal-microbiome-breast axis exists during the development of breast cancer. If so, what are the roles of host factors? Vitamin D receptor (VDR) involves host factors and the human microbiome. Vdr gene variation shapes the human microbiome, and VDR deficiency leads to dysbiosis. We hypothesized that intestinal VDR protects hosts against tumorigenesis in the breast. We examined a 7,12-dimethylbenzanthracene (DMBA)-induced breast cancer model in intestinal epithelial VDR knockout (VDRΔIEC) mice with dysbiosis. We reported that VDRΔIEC mice with dysbiosis are more susceptible to breast cancer induced by DMBA. Intestinal and breast microbiota analysis showed that VDR deficiency leads to a bacterial profile shift from normal to susceptible to carcinogenesis. We found enhanced bacterial staining within breast tumors. At the molecular and cellular levels, we identified the mechanisms by which intestinal epithelial VDR deficiency led to increased gut permeability, disrupted tight junctions, microbial translocation, and enhanced inflammation, thus increasing tumor size and number in the breast. Furthermore, treatment with the beneficial bacterial metabolite butyrate or the probiotic Lactobacillus plantarum reduced breast tumors, enhanced tight junctions, inhibited inflammation, increased butyryl-CoA transferase, and decreased levels of breast Streptococcus bacteria in VDRΔIEC mice. The gut microbiome contributes to the pathogenesis of diseases not only in the intestine but also in the breast. Our study provides insights into the mechanism by which intestinal VDR dysfunction and gut dysbiosis lead to a high risk of extraintestinal tumorigenesis. Gut-tumor-microbiome interactions represent a new target in the prevention and treatment of breast cancer.

ObjectiveThe mechanism by which migraines produce inner ear-related symptoms is not well understood. Previous studies have found that the latency of auditory brainstem response (ABR) in animal models of migraine has changed, but the threshold has not changed significantly. Therefore, it is necessary to establish a better animal model with both migraine and hearing loss to explore the relationship between migraine and auditory function deeply.MethodsIn this study, the rat model of migraine was induced by postauricular injection of nitroglycerin (NTG), and the effect on the auditory function of the inner ear was explored by comparing with intraperitoneal injection of nitroglycerin. The rats were given the drug repeatedly on alternate days, a total of 5 dosing, with the body weight monitored during the drug administration. The tactile threshold of the rats' forepaw was measured using von-Frey filaments and auditory function was assessed by ABR.ResultsThe results showed that the baseline tactile threshold of rats gradually decreased during the modeling process, and hyperalgesia appeared. Postauricular injection of NTG did not affect the weight gain of rats, while intraperitoneal injection of NTG showed slow or even negative weight gain. The ABR threshold of Click, 4 and 8 kHz of postauricular NTG injection rats increased, the latency was prolonged, and the ABR threshold in the right ear was higher than that in the left ear.ConclusionsWe demonstrated that postauricular injection of nitroglycerin may be safer and more effective than intraperitoneal injection of nitroglycerin in the process of creating rat migraine model without affecting the weight gain. Postauricular injection of nitroglycerin has more damage to the auditory function of rats. Therefore, the migraine model rat induced by postauricular injection of nitroglycerin may be a new model of cochlear migraine.

Backgrounds and Aims: Transfer RNA (tRNA) is the most extensively modified RNA in cells. Queuosine modification is a fundamental process for ensuring the fidelity and efficiency of translation from RNA to protein. In eukaryotes, Queuosine tRNA (Q-tRNA) modification relies on the intestinal microbial product queuine. However, the roles and potential mechanisms of Q-containing tRNA (Q-tRNA) modifications in inflammatory bowel disease (IBD) are unknown. Methods: We explored the Q-tRNA modifications and expression of QTRT1 (queuine tRNA-ribosyltransferase 1) in patients with IBD by investigating human biopsies and reanalyzing datasets. We used colitis models, QTRT1 knockout mice, organoids, and cultured cells to investigate the molecular mechanisms of Q-tRNA modifications in intestinal inflammation. Results: QTRT1 expression was significantly downregulated in ulcerative colitis and Crohn’s disease patients. The 4 Q-tRNA–related tRNA synthetases (asparaginyl-, aspartyl-, histidyl-, and tyrosyl-tRNA synthetase) were decreased in IBD patients. This reduction was further confirmed in a dextran sulfate sodium–induced colitis model and interleukin-10–deficient mice. Reduced QTRT1 was significantly correlated with cell proliferation and intestinal junctions, including downregulation of β-catenin and claudin-5 and the upregulation of claudin-2. These alterations were confirmed in vitro by deleting the QTRT1 gene from cells and in vivo using QTRT1 knockout mice. Queuine treatment significantly enhanced cell proliferation and junction activity in cell lines and organoids. Queuine treatment also reduced inflammation in epithelial cells. Moreover, altered QTRT1-related metabolites were found in human IBD. Conclusions: tRNA modifications play an unexplored novel role in the pathogenesis of intestinal inflammation by altering epithelial proliferation and junction formation. Further investigation of the role of tRNA modifications will uncover novel molecular mechanisms for the prevention and treatment of IBD.

Staphylococcus aureus (S. aureus) is a common pathogen that can cause many human diseases, such as skin infection, food poisoning, endocarditis, and sepsis. These diseases can be minor infections or life-threatening, requiring complex medical management resulting in substantial healthcare costs. Meanwhile, as the critically ignored “organ,” the intestinal microbiome greatly impacts physiological health, not only in gastrointestinal diseases but also in disorders beyond the gut. However, the correlation between S. aureus infection and intestinal microbial homeostasis is largely unknown. Here, we summarized the recent progress in understanding S. aureus infections and their interactions with the microbiome in the intestine. These summarizations will help us understand the mechanisms behind these infections and crosstalk and the challenges we are facing now, which could contribute to preventing S. aureus infections, effective treatment investigation, and vaccine development.

Polymerase chain reaction (PCR) is a widely used technique in gene expression analysis, diagnostics, and various molecular biology applications. However, the accuracy and sensitivity of PCR can be compromised by primer–template mismatches, potentially leading to erroneous results. In this study, we strategically designed 111 primer–template combinations with varying numbers, types, and locations of mismatches to meticulously assess their impact on qPCR performance while two distinctly different types of DNA polymerases were used. Notably, when a single-nucleotide mismatch occurred at the 3’ end of the primer, we observed significant decreases in the analytical sensitivity (0–4%) with Invitrogen™ Platinum™ Taq DNA Polymerase High Fidelity, while the analytical sensitivity remained unchanged with Takara Ex Taq Hot Start Version DNA Polymerase. Leveraging these findings, we designed a highly specific PCR to amplify Babesia while effectively avoiding the genetically close Theileria. Through elucidating the critical interplay between types of DNA polymerases and primer–template mismatches, this research provides valuable insights for improving PCR accuracy and performance. These findings have important implications for researchers aiming to achieve robust qPCR results in various molecular biology applications.

Ehrlichia canis, a prominent tick-borne pathogen causing canine monocytic ehrlichiosis (CME), is one of the six recognized Ehrlichia species worldwide. Despite its widespread presence in ticks and host dogs in China, comprehensive genomic information about this pathogen remains limited. This study focuses on an in-depth analysis of E. canis YZ-1, isolated and cultured from an infected dog in China. The complete genome of E. canis YZ-1 was sequenced (1,314,789 bp, 1022 genes, 29% GC content, and 73% coding bases), systematically characterizing its genomic elements and functions. Comparative analysis with representative genomes of Ehrlichia species, including E. canis strain Jake, E. chaffeensis, Ehrlichia spp., E. muris, E. ruminantium, and E. minasensis, revealed conserved genes, indicating potential evolutionary connections with E. ruminantium. The observed reduction in virulence-associated genes, coupled with a type IV secretion system (T4SS), suggests an intricate balance between pathogenicity and host adaptation. The close relationship with E. canis Jake and E. chaffeensis, alongside nuanced genomic variations with E. ruminantium and E. mineirensis, underscores the need to explore emerging strains and advancements in sequencing technologies continuously. This genetic insight opens avenues for innovative medications, studies on probiotic resistance, development of new detection markers, and progress in vaccine development for ehrlichiosis. Further investigations into the functional significance of identified genes and their role in host–pathogen interactions will contribute to a more holistic comprehension of Ehrlichia’s biology and its implications for pathogenicity and transmission.

Highlights BC-G@Fe0 nanocomposite was produced through two carbothermal reduction routes. The biomass impregnating-carbonizing route produced graphene-encapsulated iron carbide (Fe3C) nanoparticles. The pyrolyzing-biochar-impregnating-carbonizing route produced graphene-encapsulated nanoscale zero-valent iron particles. BC-G@Fe0 nanocomposites exhibited good heavy metal ion removal performance.

The purpose of this study was to measure the vestibular function and plasma fibrinogen level in the nitroglycerin (NTG)-induced chronic migraine rats, and explore the effect of defibrinogenation on migraine and associated vestibular dysfunction. The chronic migraine rat model was built by recurrent NTG injection. Batroxobin was administrated as a defibrinogenating drug. We measured the mechanical withdrawal threshold, vestibular function, and fibrinogen level of the rats 30 min before and 2 h following the model establishment, as well as 1 h after batroxobin administration. The results showed that vestibular function was impaired in NTG-induced chronic migraine rats. The fibrinogen levels were increased following repeated NTG injections. However, defibrinogenation did not affect either aggravating or alleviating mechanical hyperalgesia or vestibular dysfunction in the migraine model rats. These findings suggest that the NTG-induced chronic migraine rat model can be used for research on migraine-associated vestibular symptoms. Albeit the association between elevated fibrinogen levels and migraine attacks can be observed, the role of excessive fibrinogen in the pathogenesis of chronic migraine is yet to be determined.

In the development of colon cancer, the intestinal dysbiosis and disruption of barrier function are common manifestations. In the current study, we hypothesized that host factors, e.g., vitamin D receptor deficiency or adenomatous polyposis coli (APC) mutation, contribute to the enhanced dysbiosis and disrupted barrier in the pathogenesis of colorectal cancer (CRC). Using the human CRC database, we found enhanced tumor-invading bacteria and reduced colonic VDR expression, which was correlated with a reduction of Claudin-10 mRNA and protein. In the colon of VDRΔIEC mice, deletion of intestinal epithelial VDR led to lower protein of tight junction protein Claudin-10. Lacking VDR and a reduction of Claudin-10 are associated with an increased number of tumors in the mice without myeloid VDR. Intestinal permeability was significantly increased in the mice with myeloid VDR conditional deletion. Further, mice with conditional colonic APC mutation showed reduced mucus layer, enhanced bacteria in tumors, and loss of Claudin-10. Our data from human samples and colon cancer models provided solid evidence- on the host factor regulation of bacterial translocation and dysfunction on barriers in colonic tumorigenesis. Studies on the host factor regulation of microbiome and barriers could be potentially applied to risk assessment, early detection, and prevention of colon cancer.

The microbiota plays essential roles in health and disease, in both the intestine and the extra-intestine. Dysbiosis of the gut microbiota causes dysfunction in the intestine, which leads to inflammatory, immune, and infectious diseases. Dysbiosis is also associated with diseases beyond the intestine via microbial translocation or metabolisms. The in situ breast microbiome, which may be sourced from the gut through lactation and sexual contact, could be altered and cause breast diseases. In this review, we summarize the recent progress in understanding the interactions among the gut microbiome, breast microbiome, and breast diseases. We discuss the intestinal microbiota, microbial metabolites, and roles of microbiota in immune system. We emphasize the novel roles and mechanisms of the microbiome (both in situ and gastrointestinal sourced) and bacterial products in the development and progression of breast cancer. The intestinal microbial translocation suggests that the gut microbiome is translocated to the skin and subsequently to the breast tissue. The gut bacterial translocation is also due to the increased intestinal permeability. The breast and intestinal microbiota are important factors in maintaining healthy breasts. Micronutrition queuine (Q) is derived from a de novo synthesized metabolite in bacteria. All human cells use queuine and incorporate it into the wobble anticodon position of specific transfer RNAs. We have demonstrated that Q modification regulates genes critical in tight junctions and migration in human breast cancer cells and a breast tumor model. We further discuss the challenges and future perspectives that can move the field forward for prevention, diagnosis, and treatment of breast diseases.

The novel coronavirus Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has emerged and is responsible for the Coronavirus Disease 2019 global pandemic. Coronaviruses, including SARS-CoV-2, are strongly associated with respiratory symptoms during infection, but gastrointestinal symptoms, such as diarrhea, vomiting, nausea, and abdominal pain, have been identified in subsets of COVID-19 patients. This article focuses on gastrointestinal symptoms and pathophysiology in COVID-19 disease. Evidence suggests that the gastrointestinal tract could be a viral target for SARS-CoV-2 infection. Not only is the SARS-CoV-2 receptor ACE2 highly expressed in the GI tract and is associated with digestive symptoms, but bleeding and inflammation are observed in the intestine of COVID-19 patients. We further systemically summarize the correlation between COVID-19 disease, gastrointestinal symptoms and intestinal microbiota. The potential oral-fecal transmission of COVID-19 was supported by viral RNA and live virus detection in the feces of COVID-19 patients. Additionally, the viral balance in the GI tract could be disordered during SARS-CoV-2 infection which could further impact the homeostasis of the gut microbial flora. Finally, we discuss the clinical and ongoing trials of treatments/therapies, including antiviral drugs, plasma transfusion and immunoglobulins, and diet supplementations for COVID-19. By reviewing the pathogenesis of SARS-CoV-2 virus, and understanding the correlation among COVID-19, inflammation, intestinal microbiota, and lung microbiota, we provide perspective in prevention and control, as well as diagnosis and treatment of the COVID-19 disease.

Bovine leukemia virus (BLV) is widespread in global cattle populations, but the effects of its infection on milk quantity and quality have not been clearly elucidated in animal models. In this study, 30 healthy first-lactation cows were selected from ≈2,988 cows in a BLV-free farm with the same criteria of parity, age, lactation number, as well as milk yield, SCS, and composition (fat, protein, and lactose). Subsequently, these cows were randomly assigned to the intervention (n = 15) or control (n = 15) group, and reared in different cowsheds. Cows in the intervention group were inoculated with 1 × phosphate-buffered solution (PBS) resuspended in peripheral blood mononuclear cells (PBMC) from a BLV-positive cow, while the controls were inoculated with the inactivated PBMC from the same individual. From June 2016 to July 2021, milk weight (kg) was automatically recorded by milk sensors, and milk SCS and composition were originated from monthly performed dairy herd improvement (DHI) testing. Fluorescence resonance energy transfer (FRET)–qPCR and ELISA showed that cows in the intervention group were successfully infected with BLV, while cows in the control group were free of BLV for the entire period. At 45 days post-inoculation (DPI), the numbers of whole blood cells (WBCs) (P = 0.010), lymphocytes (LYMs) (P = 0.002), and monocytes (MNCs) (P = 0.001) and the expression levels of IFN-γ (P = 0.013), IL-10 (P = 0.031), and IL-12p70 (P = 0.008) increased significantly in the BLV infected cows compared to the non-infected. In lactation numbers 2–4, the intervention group had significantly higher overall milk yield (P < 0.001), fat (P = 0.031), and protein (P = 0.050) than the control group, while milk SCS (P = 0.038) and lactose (P = 0.036) decreased significantly. Further analysis indicated that BLV infection was associated with increased milk yield at each lactation stage in lactation numbers 3–4 (P = 0.021 or P < 0.001), but not with SCS and milk composition. Together, this 4-year longitudinal study revealed that artificial inoculation of BLV increased the milk yield in cows in this BLV challenge model.

Abstract The static lateral load–slip behavior of a single-shear plywood-to-plywood single-staple connection (SPSC) was investigated experimentally. A mechanics-based approach was used to develop mechanical models for deriving estimation equations for critical lateral loads of SPSCs based on failure modes of staple legs and connection member materials developed during static lateral loading process. Experimental results indicated that the static lateral load–slip behavior of SPSCs can be characterized with three major stages. This experiment provided the evidence that the ultimate lateral load capacity of SPSCs was partially governed by staple direct withdrawal load capacity in main members. The proposed mechanical models were verified experimentally as a valid means for deriving estimation equations for critical lateral loads of SPSCs evaluated in this study.

Purpose: Brain areas frequently implicated in language recovery after stroke comprise perilesional sites in the left hemisphere and homotopic regions in the right hemisphere. However, the neuronal mechanisms underlying language restoration are still largely unclear. Methods and materials: In the present study, we investigated the brain function in 15 patients with poststroke aphasia and 30 matched control subjects by combining the regional homogeneity (ReHo) and amplitudes of low-frequency fluctuation (ALFF) analysis methods based on resting-state fMRI. Results: Compared to the control subjects, the patients with aphasia exhibited increased ReHo and ALFF values in the ipsilateral perilesional areas and increased ReHo in the contralesional right middle frontal gyrus. Conclusions: The increased spontaneous brain activity in patients with poststroke aphasia during the recovery period, specifically in the ipsilateral perilesional regions and the homologous language regions of the right hemisphere, has potential implications for the treatment of patients with aphasia.

Abstract With PCR becoming one of the most important and widely-used diagnostic tools for infectious diseases of poultry, an urgent need has developed for an endogenous internal control (EIC) that monitors the quality and quantity of poultry DNA in test samples. In this study we developed a SYBR-qPCR to target the poultry homolog of the hydroxymethylbilane synthase (HMBS) gene as an EIC for avian species. The avian HMBS-based qPCR was very sensitive, detecting one HMBS gene copy in a 20 µL reaction, and is highly specific for avian species. It amplified DNA from 11 organs and tissues of chickens showing it can be used as an EIC on a large variety of samples. The application of the established EIC on clinically and experimentally infected samples demonstrated that false negativity and result variations could result from samples being collected using different operators, techniques, preservatives, and storage times. The high sensitivity and specificity of the avian HMBS-based qPCR, its ability to quantify DNAs extracted from a wide range of tissues and poultry species along with its usefulness in reducing false negativity in PCR results associated with inadequate sampling and storage degradation makes it an ideal EIC for poultry DNA and RNA PCR diagnostics. The study also highlights the importance of appropriate sampling and storage of samples in ensuring accuracy of molecular diagnostic testing.

ObjectivesThe objectives of this study were to accelerate the non-contrast-enhanced four-dimensional magnetic resonance angiography (4D MRA) based on pseudocontinuous arterial spin labeling combined with the Keyhole and View-sharing (4D-PACK) procedure using the Compressed SENSE (C-SENSE) and to improve intracranial vasculopathy evaluations for clinical purposes.Methods4D-PACK acquisition with different C-SENSE and SENSE acceleration factors was performed on 29 healthy volunteers and six patients by means of a 3.0 T MR system. Two radiologists used a 4-grade scale to qualitatively assess the vessel visualization of the middle cerebral artery (MCA) and used a 5-grade scale to qualitatively examine the image quality of 4D-PACK axial source images. Interobserver agreement was assessed by determining the weighted kappa statistic. The contrast-to-noise ratio (CNR) and arterial transmit time (ATT) were calculated in four segments of the MCA. The repeated measures one-way ANOVA for CNR and the Friedman test for source images and vessel visualization were used to analyse the differences in five sequences.Results(1) At the M4 segment, C-SENSE5 acquisition (scores, 2.72 ± 0.53) and C-SENSE6.5 (scores, 2.55 ± 0.57) provided similar vessel visualization compared with SENSE4.5 (scores, 2.72 ± 0.46); however, C-SENSE8 (scores, 1.79 ± 0.49) and C-SENSE10 (scores, 1.52 ± 0.51) had lower scores (P < 0.050). (2) The source image quality of C-SENSE5 (scores, 4.55 ± 0.51), C-SENSE6.5 (scores, 4.03 ± 0.33), and C-SENSE8 (scores, 3.48 ± 0.51) acquisition was higher than that of SENSE4.5 (scores, 3.07 ± 0.26) (P < 0.001). (3) CNRs of different MCA segments for C-SENSE5 and C-SENSE6.5 acquisitions were not significantly different compared with that of SENSE4.5 acquisition. However, the CNRs were significantly lower for C-SENSE8 (M1: 45.85 ± 13.91, M2: 27.08 ± 9.92, M4: 7.93 ± 4.49) and C-SENSE10 (M1: 37.94 ± 9.92, M2: 23.51 ± 9.0, M4: 6.78 ± 4.12) than for SENSE4.5 (M1: 55.49 ± 13.39, M2: 36.94 ± 11.02, M4: 10.18 ± 5.15) in each corresponding segment (P < 0.050). ATTs in all MCA segments within different accelerating C-SENSE factors were obviously correlated with SENSE4.5.ConclusionC-SENSE6.5 acquisition could be used to evaluate both the intracranial macrovascular and distal arteries, which could reduce the acquisition time by 18% (5 min 5 s) compared with SENSE4.5. Moreover, C-SENSE8 acquisition (37% acceleration, 3 min 54 s) could be used for routine screening and clinical diagnosis of intracranial macrovascular disease with balanced image quality.

Abstract Background Migraine is a severe and disabling brain disorder, and the exact neurological mechanisms remain unclear. Migraineurs have altered pain perception, and headache attacks disrupt their sensory information processing and sensorimotor integration. The altered functional connectivity of sub-regions of sensorimotor brain areas with other brain cortex associated with migraine needs further investigation. Methods Forty-eight migraineurs without aura during the interictal phase and 48 age- and sex-matched healthy controls underwent resting-state functional magnetic resonance imaging scans. We utilized seed-based functional connectivity analysis to investigate whether patients exhibited abnormal functional connectivity between sub-regions of sensorimotor brain areas and cortex regions. Results We found that patients with migraineurs without aura exhibited disrupted functional connectivities between the sensorimotor areas and the visual cortex, temporal cortex, posterior parietal lobule, prefrontal areas, precuneus, cingulate gyrus, sensorimotor areas proper and cerebellum areas compared with healthy controls. In addition, the clinical data of the patients, such as disease duration, pain intensity and HIT-6 score, were negatively correlated with these impaired functional connectivities. Conclusion In patients with migraineurs without aura, the functional connectivities between the sensorimotor brain areas and other brain regions was reduced. These disrupted functional connectivities might contribute to abnormalities in visual processing, multisensory integration, nociception processing, spatial attention and intention and dysfunction in cognitive evaluation and modulation of pain. Recurrent headache attacks might lead to the disrupted network between primary motor cortex and temporal regions and between primary somatosensory cortex and temporal regions. Pain sensitivity and patient quality of life are closely tied to the abnormal functional connectivity between sensorimotor regions and other brain areas.

Background: Plasmodium relictum is one of the most important avian malaria species which is mainly seen in wild birds, with infections reported in more than 70 different species and at high prevalence. Aim: The aim of this study was to determine the molecular prevalence of Plasmodium spp. in mosquitoes collected in China. Method: A Plasmodium -specific fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) was established in this study to analyze five species of mosquitoes (1,620 Culex pipiens pallens, 806 Aedes albopictus, 377 Armigeres subalbatus, 168 Anopheles sinensis, and 80 Culex tritaeniorhynchus) collected in hand nets from homes in 25 provinces of China. Results: Only females originated from six provinces were determined to be positive (0.6%, 10/1,809). Plasmodium species were detected in three mosquito species, Cx. pipiens pallens (0.5%, 8/1,620), An. sinensis (0.6%, 1/168) and Ar. subalbatus (0.3%, 1/377). Of the three mosquito species positive for P. relictum, only Cx. pipiens pallens is known to feed on birds and is recognized as the natural vector of P. relictum. Conclusion: This is the first time that P. relictum has been detected in An. sinensis and Ar. subalbatus. P. relictum, the agent of avian malaria, was present in mosquitoes in China, including in mosquito species not previously thought to be the vectors.

Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China. Author summary The mosquitoes Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus are native to China and the major vectors in the transmission of arboviruses, protozoans and nematodes. Recently, an innovative biocontrol strategy has been developed and evaluated based on the ability of Wolbachia to induce cytoplasmic incompatibility (CI), as well as interfere with the infection and replication of pathogens. Since the ability to induce CI largely depends on the density and diversity of Wolbachia, we investigated and characterized the natural infection of Wolbachia in above-mentioned five species of field-collected mosquitoes in 25 provinces of China. The results showed that the positive rates of Wolbachia infection were high in mosquitoes of Ae. albopictus, Ar. subalbatus and Cx. pipiens in large-scale regions of China and low in Cx. tritaeniorhynchus in Guizhou province. Phylogenetic analysis based on Wolbachia surface protein (wsp) gene and five multilocus sequence typing (MLST) loci indicated the high polymorphism of Wolbachia in Ae. albopictus, and low polymorphisms in Ar. subalbatus and Cx. pipiens. This finding contributes to the understanding of the nationwide distribution of Wolbachia and the potential application of this biocontrol strategy in China.

ObjectivesTo explore the differences between intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) and diffusion-weighted imaging (DWI) in evaluating the histopathological characters of pancreatic ductal adenocarcinoma (PDAC).MethodsThis retrospective study enrolled 50 patients with PDAC confirmed by pathology from December 2018 to May 2020. All patients underwent DWI and IVIM-DWI before surgeries. Patients were classified into low- and high-fibrosis groups. Apparent diffusion coefficient (ADC), diffusion coefficient (D), false diffusion coefficient (D*), and perfusion fraction (f) were measured by two radiologists, respectively in GE AW 4.7 post-processing station, wherein ADC values were derived by mono-exponential fits and f, D, D* values were derived by biexponential fits. The tumor tissue was stained with Sirius red, CD34, and CK19 to evaluate fibrosis, microvascular density (MVD), and tumor cell density. Furthermore, the correlation between ADC, D, D*, and f values and histopathological results was analyzed.ResultsThe D values were lower in the high-fibrosis group than in the low-fibrosis group, while the f values were opposite. Further, no statistically significant differences were detected in ADC and D* values between the high- and low-fibrosis groups. The AUC of D and f values had higher evaluation efficacy in the high- and low-fibrosis groups than ADC values. A significant negative correlation was established between D values, and fibrosis and a significant positive correlation were observed between f values and fibrosis. No statistical difference was detected between DWI/IVIM parameters values and MVD or tumor cell density except for the positive correlation between D* values and tumor cell density.ConclusionsD and f values derived from the IVIM model had higher sensitivity and diagnostic performance for grading fibrosis in PDAC compared to the conventional DWI model. IVIM-DWI may have the potential as an imaging biomarker for predicting the fibrosis grade of PDAC.

Abstract Background Increasing evidence has suggested that the cerebellum is associated with pain and migraine. In addition, the descending pain system of the brainstem is the major site of trigeminal pain processing and modulation and has been discussed as a main player in the pathophysiology of migraine. Cerebellar and brainstem structural changes associated with migraineurs remain to be further investigated. Methods Voxel-based morphometry (VBM) (50 controls, 50 migraineurs without aura (MWoAs)) and diffusion tensor imaging (DTI) (46 controls, 46 MWoAs) were used to assess cerebellum and brainstem anatomical alterations associated with MWoAs. We utilized a spatially unbiased infratentorial template toolbox (SUIT) to perform cerebellum and brainstem optimized VBM and DTI analysis. We extracted the average diffusion values from a probabilistic cerebellar white matter atlas to investigate whether MWoAs exhibited microstructure alterations in the cerebellar peduncle tracts. Results MWoAs showed decreased fractional anisotropy (FA) in the vermis VI extending to the bilateral lobules V and VI of the cerebellum. We also found higher axial diffusivity (AD), mean diffusivity (MD), and radial diffusivity (RD) in the right inferior cerebellum peduncle tract in MWoAs. MWoAs exhibited both reduced gray matter volume and increased AD, MD and RD in the spinal trigeminal nucleus (SpV). Conclusion MWoAs exhibited microstructural changes in the cerebellum and the local brainstem. These structural differences might contribute to dysfunction of the transmission and modulation of noxious information, trigeminal nociception, and conduction and integration of multimodal information in MWoAs. These findings further suggest involvement of the cerebellum and the brainstem in the pathology of migraine without aura.

Abstract Background Sorting Nexin 27 (SNX27) belongs to a family of sortin nexins and possesses a unique binding domain at the C-terminus which mediates protein-protein interaction in intracellular trafficking, membrane remodeling, organelle motility, and tight junctions. However, its role in cancer development, especially in vivo, remains largely unknown. Methods We have generated a stable SNX27 knockdown clone in a highly aggressive breast cancer cell line MDA-MB-231 using an inducible lentiviral shRNA system. Cell migration and proliferation of SNX27 knockdown (KD) cells were compared with wild-type (WT) cells by MTT and wound healing assay, respectively. The differences in colony formation between SNX27-KD and WT cells were detected by soft agar culture and matrigel 3D culture. Furthermore, tumor growth was examined in a xenograft nude mouse model using SNX27-KD and WT MDA-MB-231 cells. The critical EMT (epithelial-mesenchymal transition) regulators were examined in vitro and in vivo. Results The wound healing assay showed that SNX27 knockdown significantly decreased cell motility and proliferation. Colony formation in soft agar showed that the SNX27 knockdown cells formed significantly fewer and smaller colonies than the parental MDA-MB-231 cells. Western blots and immunostaining showed that knockdown of SNX27 led to increased expression of E-cadherin and β-catenin proteins, which facilitate adhesion formation and reverse EMT. EMT is a cellular program that allows polarized, immotile epithelial cells to convert to motile mesenchymal cells, promoting carcinoma invasion. The expression levels of Vimentin, the transcription factor of EMT, and tight junction protein Claudin-5, were significantly diminished in the SNX27 knockdown cells. The expression of PCNA, the cell proliferation marker, was increased in SNX27-KD cells transfected with E-cadherin siRNA. In a xenograft nude mouse model, we found that knockdown of SNX27 significantly inhibited tumor growth. The tumors from mice with SNX27-KD cells showed less proliferation compared to tumors from mice injected with wildtype cells. The increase in E-cadherin and β-catenin and decrease in Vimentin and Claudin-5 were observed in tumors of mice injected with SNX27-KD cells. Conclusions Our data have demonstrated that SNX27 plays a crucial role in tumor growth in vitro and in vivo.

Cellulose nanocrystal (CNC) has been applied in various fields due to its nano-structure, high aspect ratio, specific surface area and modulus, and abundance of hydroxy groups. In this work, CNC suspensions with different concentrations (0.4, 0.6, and 0.8%) were used as the adjuvant to improve the dispersion ability of multilayer graphene (MLG) in aqueous suspension, which is easy to be aggregated by van der Waals force between layers. In addition, N-methyl-2-pyrrolidone, ethanol, and ultrapure water were used as control groups. Zeta potential analysis and Fourier transform infrared spectroscopy showed that the stability of MLG/CNC has met the requirement, and the combination of CNC and MLG was stable in aqueous suspension. Results from transmission electron microscopy, Fourier transform infrared spectroscopy, and absorbance showed that MLG had a better dispersion performance in CNC suspensions, compared to the other solutions. Raman spectrum analysis showed that the mixtures of 1.0 wt% MLG with 0.4% CNC had the least defects and fewer layers of MLG. In addition, it is found that CNC suspension with 0.8% concentration showed the highest ability to disperse 1.0 wt% MLG with the most stable performance in suspension. Overall, this work proved the potential application of CNC as adjuvant in the field of graphene nanomaterials.

Mortality and morbidity from SARS-CoV2 (COVID-19) infections in children remains low, including an exceedingly low rate of horizontal and vertical transmission. However, unforeseen complications to childhood health have emerged secondary to the pandemic. Few studies to date have examined unintended complications of the pandemic in newborns and infants. In this Commentary, we discuss the impact that COVID-19 may have on inheritance of the newborn microbiome and its assembly throughout the first years of life. In the early stages of the pandemic when vertical transmission of COVID-19 was poorly understood, several studies reported increased rates of C-sections in COVID-19 positive women. Initial recommendations discouraged COVID-19 positive mothers from breastfeeding and participating in skin-to-skin care, advising them to isolate during their window of infectivity. These shifts in perinatal care can adversely impact microbial colonization during the first 1000 days of life. While obstetrical and neonatal management have evolved to reflect our current knowledge of perinatal transmission, we are observing other changes in early life exposures of infants, including increased attention to hygiene, fewer social interactions, and decreased global travel, all of which are major drivers of early-life gut colonization. Composition of the gut microbiota in adults directly impacts severity of infection, suggesting a role of microbial communities in modulating immune responses to COVID-19. Conversely, the role of the intestinal microbiome in susceptibility and severity of COVID-19 in newborns and children remains unknown. The onset of adulthood diseases is related to the establishment of a healthy gut microbiome during childhood. As we continue to define COVID-19 biology, further research is necessary to understand how acquisition of the neonatal microbiome is affected by the pandemic. Furthermore, infection control measures must be balanced with strategies that promote microbial diversity to impart optimal health outcomes and potentially modulate susceptibility of children to COVID-19.

Vitamin D receptor (VDR) deficiency is associated with cancer, infection, and chronic inflammation. Prior research has demonstrated VDR regulation of bacteria; however, little is known regarding VDR and viruses. We hypothesize that VDR deficiency impacts on the intestinal virome and viral-bacterial interactions. We specifically deleted VDR from intestinal epithelial cells (VDRΔIEC), Paneth cells (VDRΔPC), and myeloid cells (VDRΔLyz) in mice. Feces were collected for shotgun metagenomic sequencing and metabolite profiling. To test the functional changes, we evaluated pattern recognition receptors (PRRs) and analyzed microbial metabolites. Vibrio phages, Lactobacillus phages, and Escherichia coli typing phages were significantly enriched in all three conditional VDR-knockout mice. In the VDRΔLyz mice, the levels of eight more virus species (2 enriched, 6 depleted) were significantly changed. Altered virus species were primarily observed in female VDRΔLyz (2 enriched, 3 depleted) versus male VDRΔLyz (1 enriched, 1 depleted). Altered alpha and beta diversity (family to species) were found in VDRΔLyz. In VDRΔIEC mice, bovine viral diarrhea virus 1 was significantly enriched. A significant correlation between viral and bacterial alterations was found in conditional VDR knockout mice. There was a positive correlation between Vibrio phage JSF5 and Cutibacterium acnes in VDRΔPC and VDRΔLyz mice. Also, there were more altered viral species in female conditional VDR knockout mice. Notably, there were significant changes in PRRs: upregulated TLR3, TLR7, and NOD2 in VDRΔLyz mice and increased CLEC4L expression in VDRΔIEC and VDRΔPC mice. Furthermore, we identified metabolites related to virus infection: decreased glucose in VDRΔIEC mice, increased ribulose/xylulose and xylose in VDRΔLyz mice, and increased long-chain fatty acids in VDRΔIEC and VDRΔLyz female mice. Tissue-specific deletion of VDR changes the virome and functionally changes viral receptors, which leads to dysbiosis, metabolic dysfunction, and infection risk. This study helps to elucidate VDR regulating the virome in a tissue-specific and sex-specific manner.

Infectious agents, including viruses, bacteria, fungi, and parasites, have been linked to pathogenesis of human cancers, whereas viruses and bacteria account for more than 99% of infection associated cancers. The human microbiome consists of not only bacteria, but also viruses and fungi. The microbiome co-residing in specific anatomic niches may modulate oncologic potentials of infectious agents in carcinogenesis. In this review, we focused on interactions between viruses and bacteria for cancers arising from the orodigestive tract and the female genital tract. We examined the interactions of these two different biological entities in the context of human carcinogenesis in the following three fashions: (1) direct interactions, (2) indirect interactions, and (3) no interaction between the two groups, but both acting on the same host carcinogenic pathways, yielding synergistic or additive effects in human cancers, e.g., head and neck cancer, liver cancer, colon cancer, gastric cancer, and cervical cancer. We discuss the progress in the current literature and summarize the mechanisms of host-viral-bacterial interactions in various human cancers. Our goal was to evaluate existing evidence and identify gaps in the knowledge for future directions in infection and cancer.

Tensile fatigue performances of selected natural rattan strips (NRSs) and synthetic rattan strips (SRSs) were evaluated by subjecting them to zero-to-maximum constant amplitude cyclic tensile loading. Experimental results indicated that a fatigue life of 25,000 cycles began at the stress level of 50% of rattan material ultimate tensile strength (UTS) value for NRSs evaluated. Rattan core strips’ fatigue life of 100,000 cycles started at the stress level of 30% of its UTS value. Rattan bast strips could start a fatigue life of 100,000 cycles at a stress level below 30% of material UTS value. SRSs didn’t reach the fatigue life of 25,000 cycles until the applied stress level reduced to 40% of material UTS value and reached the fatigue life of 100,000 cycles at the stress level of 40% of material UTS value. It was found that NRSs’ S-N curves (applied nominal stress versus log number of cycles to failure) could be approximated by S=σou(1−H×log10⋅Nf). The constant H values in the equation were 0.10 and 0.08 for bast and core materials, respectively.

Abstract Phenotypic determination of antimicrobial resistance in bacteria is very important for diagnosis and treatment, but sometimes this procedure needs further genetic evaluation. Whole-genome sequencing plays a critical role in deciphering and advancing our understanding of bacterial evolution, transmission, and surveillance of antimicrobial resistance. In this study, whole-genome sequencing was performed on nineteen clinically extraintestinal Escherichia coli isolates from chicken, cows and swine and showing different antimicrobial susceptibility. A total of 44 different genes conferring resistance to 11 classes of antimicrobials were detected in 15 of 19 E. coli isolates (78.9%), and 22 types of plasmids were detected in 15/19 (78.9%) isolates. In addition, whole-genome sequencing of these 19 isolates identified 111 potential virulence factors, and 53 of these VFDB-annotated genes were carried by all these 19 isolates. Twelve different virulence genes were identified while the most frequent ones were gad (glutamate decarboxylase), iss (increased serum survival) and lpfA (long polar fimbriae). All isolates harbored at least one of the virulence genes. The findings from comparative genomic analyses of the 19 diverse E. coli isolates in this study provided insights into molecular basis of the rising multi-drug resistance in E. coli.

Abstract Antimicrobial resistance against colistin has emerged worldwide and is threatening the efficacy of colistin treatment of multi-resistant Gram-negative bacteria. In this study, PCRs were used to detect mcr genes (mcr-1, mcr-2, mcr-3) in 213 anal and 1,339 nasal swabs from pigs (n = 1,454) in nine provinces of China, and 1,696 cloacal and 1,647 oropharyngeal samples from poultry (n = 1,836) at live-bird markets in 24 provinces. The mcr-1 prevalences in pigs (79.2%) and geese (71.7%) were significantly higher than in chickens (31.8%), ducks (34.6%) and pigeons (13.1%). The mcr-2 prevalence in pigs was 56.3%, significantly higher than in chickens (5.5%), ducks (2.3%), geese (5.5%) and pigeons (0%). The mcr-3 prevalences in pigs (18.7%), ducks (13.8%) and geese (11.9%) were significantly higher than in chickens (5.2%) and pigeons (5.1%). In total, 173 pigs and three chickens were positive for all three mcr genes. The prevalences of the mcr were significantly higher in nasal/oropharyngeal swabs than in the anal /cloacal swabs. Phylogenetic studies identified 33 new mcr-2 variants and 12 new mcr-3 variants. This study demonstrates high prevalences of mcr in pigs and poultry in China, and indicates there is need for more thorough surveillance and control programs to prevent further selection of colistin resistance.

Abstract Objective Colistin resistance has emerged worldwide and has been threatening the efficacy of one of the last-resort antimicrobials used for treatment of multidrug resistant Gram-negative bacteria. While five colistin resistance genes (mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5) have been described, few data are available on the prevalence of mcr-genes other than mcr-1 in human samples. Results In this study, the presence of five currently described colistin resistance genes (mcr 1–5) in vaginal swabs of women undergoing infertility evaluation was reported. Most samples were found to be positive for the mcr-4 (12.7%), followed by two for the mcr-2 (1.5%), two for the mcr-3 (1.5%), one for the mcr-1 (0.7%), and one for the mcr-5 (0.7%). Phylogenetic comparison demonstrated identical (mcr-1, mcr-2, mcr-3, mcr-5) or similar (mcr-4) nucleotide sequences of human samples and those of animal origins from the same city, suggesting the potential transmission of mcr genes from animals to humans. This is the first detection of mcr-2, mcr-4 and mcr-5 genes in human samples, and warrants further research to determine the spread of the mcr genes and elucidate the full epidemiology of colistin resistance genes in humans.

The polarization degree of objects in the marine background are affected by infrared radiation from sea surface. Taking into account the radiation coupling effect (RCE), a degree of linear polarization (DoLP) model is deduced. The DoLP of painted aluminum plates at different observation angles are simulated. The simulation results show the trend of the DoLP of the object decreases first and then increases as the observation angle θO, with the minimum value at θO=53∘. Nevertheless, we get a monotonically increasing trend and the minimum value is at θO=0∘ without considering RCE. The experimental results accord closely with those of the simulation with RCE. This conclusion is useful for the polarization detection and identification of infrared objects in the marine background.

Abstract Background Chlamydia suis is an important, globally distributed, highly prevalent and diverse obligate intracellular pathogen infecting pigs. To investigate the prevalence and genetic diversity of C. suis in China, 2,137 nasal, conjunctival, and rectal swabs as well as whole blood and lung samples of pigs were collected in 19 regions from ten provinces of China in this study. Results We report an overall positivity of 62.4% (1,334/2,137) of C. suis following screening by Chlamydia spp. 23S rRNA-based FRET-PCR and high-resolution melting curve analysis and confirmatory sequencing. For C. suis-positive samples, 33.3 % of whole blood and 62.5% of rectal swabs were found to be positive for the C. suis tetR(C) gene, while 13.3% of whole blood and 87.0% of rectal swabs were positive for the C. suis tet(C) gene. Phylogenetic comparison of partial C. suis ompA gene sequences revealed significant genetic diversity in the C. suis strains. This genetic diversity was confirmed by C. suis-specific multilocus sequence typing (MLST), which identified 26 novel sequence types among 27 examined strains. Tanglegrams based on MLST and ompA sequences provided evidence of C. suis recombination amongst the strains analyzed. Conclusions Genetically highly diverse C. suis strains are exceedingly prevalent in pigs. As it stands, the potential pathogenic effect of C. suis on pig health and production of C. suis remains unclear and will be the subject of further investigations. Further study is also required to address the transmission of C. suis between pigs and the risk of 'spill-over' and 'spill-back' of infections to wild animals and humans.

Abstract Chlamydia trachomatis, Mycoplasma spp., Neisseria gonorrhoeae and Treponema pallidum are sexually transmitted pathogens that threaten reproductive health worldwide. In this study, vaginal swabs obtained from women (n = 133) that attended an infertility clinic in China were tested with qPCRs for C. trachomatis, Mycoplasma spp., N. gonorrhoeae, T. pallidum and tetracycline resistance genes. While none of vaginal swabs were positive for N. gonorrhoeae and T. pallidum, 18.8% (25/133) of the swabs were positive for Chlamydia spp. and 17.3% of the swabs (23/133) were positive for Mycoplasma species. All swabs tested were positive for tetracycline resistance gene tet(M) which is the most effective antibiotic for bacterial sexually transmitted infections. The qPCRs determined that the gene copy number per swab for tet(M) was 7.6 times as high as that of C. trachomatis 23S rRNA, and 14.7 times of Mycoplasma spp. 16S rRNA. In China, most hospitals do not detect C. trachomatis and Mycoplasma spp. in women with sexually transmitted infections and fertility problems. This study strongly suggests that C. trachomatis and Mycoplasma spp. should be routinely tested in women with sexually transmitted infections and infertility in China, and that antimicrobial resistance of these organisms should be monitored. Further studies are warranted to determine the prevalences in different regions and associated risk factors.

Abstract Background As there is little data on vector-borne diseases of cats in the Caribbean region and even around the world, we tested feral cats from St Kitts by PCR to detect infections with Babesia, Ehrlichia and spotted fever group Rickettsia (SFGR) and surveyed them for antibodies to Rickettsia rickettsii and Ehrlichia canis. Results Whole blood was collected from apparently healthy feral cats during spay/ neuter campaigns on St Kitts in 2011 (N = 68) and 2014 (N = 52). Sera from the 52 cats from 2014 were used to detect antibodies to Ehrlichia canis and Rickettsia rickettsii using indirect fluorescent antibody tests and DNA extracted from whole blood of a total of 119 cats (68 from 2011, and 51 from 2014) was used for PCRs for Babesia, Ehrlichia and Rickettsia. We could not amplify DNA of SFG Rickettsia in any of the samples but found DNA of E. canis in 5% (6/119), Babesia vogeli in 13% (15/119), Babesia gibsoni in 4% (5/119), mixed infections with B. gibsoni and B. vogeli in 3% (3/119), and a poorly characterized Babesia sp. in 1% (1/119). Overall, 10% of the 52 cats we tested by IFA for E. canis were positive while 42% we tested by indirect fluorescent antibody (IFA) for R. rickettsii antigens were positive. Conclusions Our study provides the first evidence that cats can be infected with B. gibsoni and also indicates that cats in the Caribbean may be commonly exposed to other vector-borne agents including SFGR, E. canis and B. vogeli. Animal health workers should be alerted to the possibility of clinical infections in their patients while public health workers should be alerted to the possibility that zoonotic SFGR are likely circulating in the region.

Abstract Background Migraine constitute a disorder characterized by recurrent headaches, and have a high prevalence, a high socio-economic burden and severe effects on quality of life. Our previous fMRI study demonstrated that some brain regions are functional alterations in migraineurs. As the function of the human brain is related to its structure, we further investigated white and gray matter structural alterations in migraineurs. Methods In current study, we used surface-based morphometry, voxel-based morphometry and diffusion tensor imaging analyses to detect structural alterations of the white matter and gray matter in 32 migraineurs without aura compared with 32 age- and gender-matched healthy controls. Results We found that migraineurs without aura exhibited significantly increased gray matter volume in the bilateral cerebellar culmen, increased cortical thickness in the lateral occipital-temporal cortex, decreased cortical thickness in the right insula, increased gyrification index in left postcentral gyrus, superior parietal lobule and right lateral occipital cortex, and decreased gyrification index in the left rostral middle frontal gyrus compared with controls. No significant change in white matter microstructure was found in DTI analyses. Conclusion The significantly altered gray matter brain regions were known to be associated with sensory discrimination of pain, multi-sensory integration and nociceptive information processing and were consistent with our previous fMRI study, and may be involved in the pathological mechanism of migraine without aura.

Palm petioles fibers (PPF) are used widely in China to make mattresses. The changes of the properties of PPF after heat treatment is an important factor that should be considered. This study focuses on the fiber tensile test after heat treatment under different temperatures and at different times. As the temperature of treatment was increased, the Young’s modulus of PPF increased, while the breaking strength and elongation declined. The turning point of the tensile properties was 160 °C. The heating time also had a significant influence on the tensile properties. As the heating time increased, the Young’s modulus grew, while the breaking strength and elongation declined.

Bartonella are vector-borne hemotropic bacteria that infect a wide variety of hosts, including people. While there are PCR assays that can identify individual or groups of Bartonella, there is no reliable molecular method to simultaneously detect all species while maintaining genus specificity and sensitivity. By comparing highly conserved 16S rRNA sequences of the better-recognized Bartonella spp. on GenBank, we selected primers and probes for a genus-specific pan-Bartonella FRET-qPCR. Then, a gltA-based Bartonella PCR was established by selecting primers for a highly variable region of gltA, of which the sequenced amplicons could identify individual Bartonella spp. The pan-Bartonella FRET-qPCR did not detect negative controls (Brucella spp., Anaplasma spp., Rickettsia spp., Coxiella burnetii, and Wolbachia) but reliably detected as few as two copies of the positive control (Bartonella henselae) per reaction. There was complete agreement between the pan-Bartonella FRET-qPCR and the gltA-based Bartonella PCR in detecting Bartonella in convenience test samples from China and St. Kitts: cats (26%; 81/310), Ctenocephalides felis (20%; 12/60), cattle (24%; 23/98), and donkeys (4%; 1/20). Sequencing of the gltA-based Bartonella PCR products revealed B. henselae (70%; 57/81) and B. clarridgeiae (30%; 24/81) in cats and C. felis (67%; 8/12, and 33%; 4/12, respectively) and B. bovis in cattle (23.5%; 23/98) and donkeys (4.0%; 1/24). The pan-Bartonella FRET-qPCR and gltA-based Bartonella PCR we developed are highly sensitive and specific in detecting recognized Bartonella spp. in a single reaction. The pan-Bartonella FRET-qPCR is convenient requiring no gel electrophoresis and providing copy numbers, while the gltA-based Bartonella PCR reliably differentiates individual Bartonella species. The use of these PCRs should greatly facilitate large-scale surveillance studies and the diagnosis of infections in clinical samples.

Rickettsia felis belongs to spotted fever group Rickettsia and is an emerging human pathogen most commonly transmitted by a range of fleas and ticks. While recent evidence has suggested mosquitoes are infected with R. felis, there is little information about the role of mosquitoes in the organism’s transmission. In this study, around 100 mosquitoes were collected monthly between 2013 and 2014 from the same residential dwelling at Yangzhou, China. The collected mosquitoes were identified for their species and gender, followed by gltA-based PCR and hydroxymethylbilane synthase-based PCR to determine the prevalence of Rickettsia and blood meal. Three mosquito species (Culex pipiens: 76%, 996/1,304; C. tritaeniorhynchus: 17%, 216/1,304; Aedes albopictus: 7%, 92/1,304) were identified. For 1,088 female mosquitoes, 31% of them n=336 were positive for blood meal and 7% n=77 carried R. felis DNA. In a strong contrast, none of the 216 male mosquitoes were positive for blood meal but two males were positive for Rickettsia. Interestingly, 63% of R. felis-positive mosquitoes (50/79) were negative for blood meal, being significantly higher than 37% of mosquitoes and being positive for both R. felis and blood meal P=0.008. Furthermore, we compared the prevalence of Rickettsia and blood meal in the mosquitoes collected in the months with temperature below and above 23°C, the minimum temperature required for mosquito egg hatching. Mosquitoes captured in the months below 23°C showed significant higher positivity of R. felis(71/936, 7.6% vs. 8/368, 2.2%; P=0.002) and blood meal (294/936, 31.4% vs. 36/368, 9.8%; P

Avermectin is commonly used in aquaculture systems for pest control in recent decades in China. However, no information is provided for the toxic effect to the important commercial species, Chinese mitten crab, Eriocheir sinensis. To investigate the aquatic toxicity of avermectin, an acute toxic test was performed in this study. The results showed that the 48 h- and 96 h- LC50 were 1.663 and 0.954 mg/L, respectively. For further research, crabs were exposed to sublethal concentrations of 0.03, 0.06, 0.12, 0.24 and 0.48 mg/L. Levels of antioxidants, including superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) were significantly (P