Myeloid vitamin D receptor regulates Paneth cells and microbial homeostasis.

Cross talk between immune cells and the intestinal crypt is critical in maintaining intestinal homeostasis. Recent studies highlight the direct impact of vitamin D receptor (VDR) signaling on intestinal and microbial homeostasis. However, the tissue-specific role of immune VDR signaling is not fully understood. Here, we generated a myeloid-specific VDR knockout (VDRΔLyz ) mouse model and used a macrophage/enteroids coculture system to examine tissue-specific VDR signaling in intestinal homeostasis. VDRΔLyz mice exhibited small intestine elongation and impaired Paneth cell in maturation and localization. Coculture of enteroids with VDR-/- macrophages increased the delocalization of Paneth cells. VDRΔLyz mice exhibited significant changes in the microbiota taxonomic and functional files, and susceptibility to Salmonella infection. Interestingly, loss of myeloid VDR impaired Wnt secretion in macrophages, thus inhibiting crypt ß-catenin signaling and disrupting Paneth cell differentiation in the epithelium. Taken together, our data have demonstrated that myeloid cells regulate crypt differentiation and the microbiota in a VDR-dependent mechanism. Dysregulation of myeloid VDR led to high risks of colitis-associated diseases. Our study provided insight into the mechanism of immune/Paneth cell cross talk in regulating intestinal homeostasis.

Low phosphorus increases hepatic lipid deposition, oxidative stress and inflammatory response via Acetyl-CoA carboxylase-dependent manner in zebrafish liver cells.

Acetyl-CoA carboxylase (ACC) plays a regulatory role in both fatty acid synthesis and oxidation, controlling the process of lipid deposition in the liver. Given that existing studies have shown a close relationship between low phosphorus (P) and hepatic lipid deposition, this study was conducted to investigate whether ACC plays a crucial role in this relationship. Zebrafish liver cell line (ZFL) was incubated under low P medium (LP, P concentration: 0.77 mg/L) or adequate P medium (AP, P concentration: 35 mg/L) for 240 h. The results showed that, compared with AP-treated cells, LP-treated cells displayed elevated lipid accumulation, and reduced fatty acid ß-oxidation, ATP content, and mitochondrial mass. Furthermore, transcriptomics analysis revealed that LP-treated cells significantly increased lipid synthesis (Acetyl-CoA carboxylases (acc), Stearyl coenzyme A dehydrogenase (scd)) but decreased fatty acid ß-oxidation (Carnitine palmitoyltransferase I (cptI)) and (AMP-activated protein kinase (ampk)) mRNA levels compared to AP-treated cells. The phosphorylation of AMPK and ACC, and the protein expression of CPTI were significantly decreased in LP-treated cells compared with those in AP-treated cells. After 240 h of LP treatment, PF-05175157 (an ACC inhibitor) was supplemented in the LP treatment for an additional 12 h. PF-05175157-treated cells showed higher phosphorylation of ACC, higher protein expression of CPTI, and lower protein expression of FASN, lower TG content, enhanced fatty acid ß-oxidation, increased ATP content, and mitochondrial mass compared with LP-treated cells. PF-05175157 also relieved the LP-induced oxidative stress and inflammatory response. Overall, these findings suggest that ACC is a promising target for treating LP-induced elevation of lipid deposition in ZFL, and can alleviate oxidative stress and inflammatory response.

Lignocellulosic Biomass-Based Carbon Dots: Synthesis Processes, Properties, and Applications.

Carbon dots (CDs), a new type of carbon-based fluorescent nanomaterial, have attracted widespread attention because of their numerous excellent properties. Lignocellulosic biomass is the most abundant renewable natural resource and possesses broad potential to manufacture different composite and smart materials. Numerous studies have explored the potential of using the components (such as cellulose, hemicellulose, and lignin) in lignocellulosic biomass to produce CDs. There are few papers systemically aiming in the review of the state-of-the-art works related to lignocellulosic biomass-derived CDs. In this review, the significant advances in synthesis processes, formation mechanisms, structural characteristics, optical properties, and applications of lignocellulosic biomass-based CDs such as cellulose-based CDs, hemicellulose-based CDs and lignin-based CDs in latest research are reviewed. In addition, future research directions on the improvement of the synthesis technology of CDs using lignocellulosic biomass as raw materials to enhance the properties of CDs are proposed. This review will serve as a road map for scientists engaged in research and exploring more applications of CDs in different science fields to achieve the highest material performance goals of CDs.

Risk Factors for Cerebrovascular Events in Moyamoya Angiopathy Using 4D Flow MRI: A Pilot Study.

BACKGROUND: Little is known about internal carotid artery (ICA) hemodynamics in patients with moyamoya angiopathy (MMA) and its role in cerebrovascular events. PURPOSE: To characterize ICA hemodynamics in MMA patients by 4D flow MRI and investigate its relationship with cerebrovascular events. STUDY TYPE: Prospective. SUBJECTS: Seventy MMA patients (50 years old ± 9, 30 males). FIELD STRENGTH/SEQUENCE: Time-resolved three-directional velocity encoded fast field echo sequence (4D flow) MRI, T1-weighted fast field echo sequence, T2 weighted turbo spin echo sequence, diffusion weighted echo planar imaging; T2-weighted fluid-attenuated inversion recovery turbo spin echo sequence, susceptibility weighted fast field echo sequence, and time-of-flight MR angiography fast field echo sequence at 3.0T. ASSESSMENT: ICA hemodynamics (maximum and average velocity [Vmax , Vavg ], average blood flow [Flowavg ], and wall shear stress) were analyzed based on 4D flow data. Cerebral infarction, defined as the occurrence of events, in 124 brain hemispheres was determined according to clinical symptoms and conventional brain MR imaging. STATISTICAL TESTS: The independent-samples T test was used to evaluate differences in ICA hemodynamics between infarcted and non-infarcted hemispheres. Binary logistic regression was performed to investigate the relationship between ICA hemodynamics and events. A P value < 0.05 was considered statistically significant. RESULTS: Sixty-one infarcted hemispheres (eight hemispheres with acute ischemic damage, 30 with chronic ischemic damage, and 23 with chronic hemorrhagic damage) had cerebrovascular events and 63 non-infarcted hemispheres did not. The hemodynamic parameters in the infarcted hemispheres (Vmax : P < 0.001; Vavg : P = 0.003; and Flowavg : P = 0.004) were significantly lower than those in the non-infarcted hemispheres. However, Vmax (P = 0.052), Vavg (P = 0.107), and Flowavg (P = 0.074) were not significantly different among hemispheres with acute ischemic lesions, chronic ischemic lesions and chronic hemorrhagic lesions. Vmax (odds ratio 3.033, 95% CI: 1.075-8.562) was independently associated with cerebrovascular events. DATA CONCLUSION: Vmax maybe a higher risk factor of cerebrovascular events in MMA patients. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY STAGE: 3.

A deep learning method for the automated assessment of paradoxical pulsation after myocardial infarction using multicenter cardiac MRI data.

OBJECTIVE: The current study aimed to explore a deep convolutional neural network (DCNN) model that integrates multidimensional CMR data to accurately identify LV paradoxical pulsation after reperfusion by primary percutaneous coronary intervention with isolated anterior infarction. METHODS: A total of 401 participants (311 patients and 90 age-matched volunteers) were recruited for this prospective study. The two-dimensional UNet segmentation model of the LV and classification model for identifying paradoxical pulsation were established using the DCNN model. Features of 2- and 3-chamber images were extracted with 2-dimensional (2D) and 3D ResNets with masks generated by a segmentation model. Next, the accuracy of the segmentation model was evaluated using the Dice score and classification model by receiver operating characteristic (ROC) curve and confusion matrix. The areas under the ROC curve (AUCs) of the physicians in training and DCNN models were compared using the DeLong method. RESULTS: The DCNN model showed that the AUCs for the detection of paradoxical pulsation were 0.97, 0.91, and 0.83 in the training, internal, and external testing cohorts, respectively (p < 0.001). The 2.5-dimensional model established using the end-systolic and end-diastolic images combined with 2-chamber and 3-chamber images was more efficient than the 3D model. The discrimination performance of the DCNN model was better than that of physicians in training (p < 0.05). CONCLUSIONS: Compared to the model trained by 2-chamber or 3-chamber images alone or 3D multiview, our 2.5D multiview model can combine the information of 2-chamber and 3-chamber more efficiently and obtain the highest diagnostic sensitivity. CLINICAL RELEVANCE STATEMENT: A deep convolutional neural network model that integrates 2-chamber and 3-chamber CMR images can identify LV paradoxical pulsation which correlates with LV thrombosis, heart failure, ventricular tachycardia after reperfusion by primary percutaneous coronary intervention with isolated anterior infarction. KEY POINTS: • The epicardial segmentation model was established using the 2D UNet based on end-diastole 2- and 3-chamber cine images. • The DCNN model proposed in this study had better performance for discriminating LV paradoxical pulsation accurately and objectively using CMR cine images after anterior AMI compared to the diagnosis of physicians in training. • The 2.5-dimensional multiview model combined the information of 2- and 3-chamber efficiently and obtained the highest diagnostic sensitivity.

Paneth Cell Alertness to Pathogens Maintained by Vitamin D Receptors.

BACKGROUND AND AIMS: Vitamin D exerts a regulatory role over mucosal immunity via the vitamin D receptor (VDR). Although Paneth cells and their products are known to regulate the commensal and pathogenic microbiota, the role that VDRs in Paneth cells play in these responses is unknown. METHODS: We identified the decreased intestinal VDR significantly correlated with reduction of an inflammatory bowel disease risk gene ATG16L1 and Paneth cell lysozymes in patients with Crohn's disease. We generated Paneth cell-specific VDR knockout (VDRΔPC) mice to investigate the molecular mechanisms. RESULTS: Lysozymes in the Paneth cells were significantly decreased in the VDRΔPC mice. Isolated VDRΔPC Paneth cells exhibited weakened inhibition of pathogenic bacterial growth and displayed reduced autophagic responses. VDRΔPC mice had significantly higher inflammation after Salmonella infections. VDRΔPC mice also showed high susceptibility to small intestinal injury induced by indomethacin, a nonsteroidal anti-inflammatory drug. Co-housing of VDRΔPC and VDRlox mice made the VDRΔPC less vulnerable to dextran sulfate sodium colitis, suggesting the transmission of protective bacterial from the VDRlox mice. Thus, a lack of VDR in Paneth cells leads to impaired antibacterial activities and consequently increased inflammatory responses. Genetically and environmentally regulated VDRs in the Paneth cells may set the threshold for the development of chronic inflammation, as observed in inflammatory bowel diseases. CONCLUSIONS: We provide new insights into the tissue-specific functions of VDRs in maintaining Paneth cell alertness to pathogens in intestinal disorders. Targeting the VDR affects multiple downstream events within Paneth cells that inhibit intestinal inflammation and establish host defense against enteropathogens.

Vessel-selective 4D MRA based on ASL might potentially show better performance than 3D TOF MRA for treatment evaluation in patients with intra-extracranial bypass surgery: a prospective study.

OBJECTIVES: To compare bypass patency and intracranial collaterals from the external carotid artery (ECA) by four-dimensional MR angiography (MRA) based on super-selective pseudo-continuous arterial spin labeling (pCASL) combined with the keyhole and view-sharing techniques (4D-sPACK) versus three-dimensional time-of-flight (3D TOF) MRA in patients with extra-intracranial revascularization. METHODS: The MR data of 45 patients administered bypass surgery were collected. The image quality of 4D-sPACK was evaluated using a 4-point grading system according to whether the diagnosis of intracranial collaterals was affected. Anastomosis patency and intracranial collateral visualization from ECA were assessed by two radiologists on 4D-sPACK and 3D TOF MRA, with digital subtraction angiography (DSA) findings as reference. Intracranial collateral assessment employed another 4-point grading system according to the number of vessels shown. Interobserver agreement was assessed with the weighted kappa statistic. RESULTS: Fifty hemispheres in 43 patients were included. The image quality of 4D-sPACK was good in 47 (47/50, 94.0%) hemispheres. 4D-sPACK had a higher sensitivity than 3D TOF MRA (97.73% vs 79.55%) for visualizing anastomoses. There were significant differences between 4D-sPACK (scores, 3.22 ± 1.15) and 3D TOF MRA (scores, 1.80 ± 0.67) in the visualization of intracranial collaterals from ECA (p < 0.001). The interobserver agreement was substantial for intracranial collateral assessment (κ4D-sPACK = 0.788; κ3D TOF MRA = 0.800) and almost perfect for bypass patency (κ4D-sPACK = 0.912; κ3D TOF MRA = 0.816; κDSA = 0.811). CONCLUSION: This pilot study shows that, 4D-sPACK has a better performance than 3D TOF MRA in treatment evaluation of patients after bypass surgery, and has high consistency with DSA. KEY POINTS: • 4D-sPACK is a non-contrast-enhanced dynamic MRA method for the visualization of intracranial vessels. • 4D-sPACK has higher specificity for the diagnosis of anastomosis occlusion. • 4D-sPACK is better than 3D TOF MRA in the visualization of intracranial collaterals in patients after bypass surgery.

Long-Term Prognosis of Tinnitus Associated with Idiopathic Sudden Sensorineural Hearing Loss.

OBJECTIVE: To analyze the factors affecting the long-term prognosis of tinnitus accompanied by unilateral idiopathic sudden sensorineural hearing loss (SSNHL). METHODS: A total of 161 patients with sudden hearing loss (HL) accompanied by tinnitus were enrolled. All patients had two separate telephone follow-ups and were asked about changes in tinnitus. The severity of tinnitus at admission and the outcome at discharge were assessed in terms of the patients' sex, age, level of HL, type of audiogram, etc. Results: The prognosis of tinnitus after SSNHL had no relationship with grades of HL or hearing recovery. Initial tinnitus level was remarkably associated with tinnitus improvement at discharge and was an independent risk factor for the long-term prognosis of residual tinnitus after SSNHL (odds ratio 0.722, 95% confidence interval 0.550-0.949, p = 0.019), and the median recovery time was 23.00 ± 3.80 months. CONCLUSIONS: Residual tinnitus after SSNHL has a tendency of self-recovery. The short-term prognosis of tinnitus may be related to psychological changes caused by hearing recovery, while the long-term prognosis of residual tinnitus after SSNHL is related only to the initial tinnitus level, with a median recovery time of approximately 2 years.

Disrupted functional connectivity between sub-regions in the sensorimotor areas and cortex in migraine without aura.

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.

Deletion of sorting nexin 27 suppresses proliferation in highly aggressive breast cancer MDA-MB-231 cells in vitro and in vivo.

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 ß-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 ß-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.

Antioxidative status, immunological responses, and heat shock protein expression in hepatopancreas of Chinese mitten crab, Eriocheir sinensis under the exposure of glyphosate.

As a broad-spectrum herbicide, glyphosate was extensively utilised in China for several decades. The contradiction between glyphosate spraying and crab breeding in the rice-crab co-culture system has become more obvious. In this study, the antioxidative status and immunological responses of Chinese mitten crab, Eriocheir sinensis, under sublethal exposure of glyphosate were investigated by detecting the antioxidative and immune-related enzyme activity, acetylcholinesterase (AChE) activity and relative mRNA expression of heat shock proteins (HSPs) in hepatopancreas. The results showed that high concentrations of glyphosate (44 and 98 mg/L) could induce significant alteration of superoxide dismutase (SOD), peroxidase (POD), acid phosphatase (ACP), alkaline phosphatase (AKP), and phenoloxidase (PO) activities by first rising then falling during the exposure. However, AChE activity in all treatments including 4.4 mg/L was inhibited markedly after 6 h of exposure. In addition, the relative mRNA expression of HSP 60, HSP 70, and HSP 90 was significantly upregulated at both 48 h and 96 h. These results revealed that glyphosate has a prominent toxic effect on E. sinensis based on antioxidative and immunological response inhibition and AChE activity reduction even at the lowest concentration of 4.4 mg/L, and a protective response by upregulation of HSPs was carried out by the species to ease the environmental stress.

Abnormal neural responses to emotional stimuli in children with primary monosymptomatic nocturnal enuresis.

Nocturnal enuresis (NE) is a common disorder in school-aged children that has been reported to affect nearly 10% of 7-year-old children and affects both the children and their families. Previous studies have shown that the risk of psychosocial difficulties in children with enuresis is elevated. Thus, children with NE may experience negative effects on psychosocial health or emotion processing. Therefore, the aim of this study was to investigate the potential disturbance in emotional processing in children with NE using functional magnetic resonance imaging (fMRI). In this work, we used fMRI and an affective picture task to evaluate brain response changes in children with NE. Two groups, one consisting of 22 children with primary monosymptomatic NE and one with 23 healthy controls, were scanned using fMRI. Compared to the healthy subjects, children with NE mainly showed increased activation when viewing negative vs. neutral pictures in the bilateral medial superior frontal gyrus that extended to the anterior cingulate cortex. Our results demonstrated that children with primary monosymptomatic NE showed abnormal neural responses to emotional stimuli and overactivation in the medial prefrontal and anterior cingulate cortices suggested that children with primary monosymptomatic NE may be hypersensitive in their sensory perception of negative pictures.

Brain Structural Plasticity Associated with Maternal Caregiving in Mothers: A Voxel- and Surface-Based Morphometry Study.

BACKGROUND: Pregnancy constitutes a significant period in the lives of women, after which they often experience numerous crucial physiological and psychological changes. Functional neuroimaging studies have shown longitudinal changes in functional brain activity in mothers responding to infant-related stimuli. However, the structural changes that occur in the brains of mothers after delivery remain to be explored. OBJECTIVE: We aimed to evaluate the structural changes in mothers during the postpartum phase. METHODS: We recruited 35 primiparous mothers and 26 nonmothers to participate in this voxel- and surface-based morphometry study, and 22 mothers were scanned twice with a follow-up of approximately 2 years. RESULTS: Compared to nonmothers, mothers exhibited reduced gray matter (GM) volumes and increased white matter (WM) volumes in regions associated with empathy and reward networks (supplementary motor area, precuneus, inferior parietal lobe, insula, and striatum), decreased cortical thickness in the precentral gyrus and increased gyrification index in the orbitofrontal cortex. Furthermore, mothers showed longitudinal changes in the GM and WM volumes and cortical thickness of several of these regions (including the superior and medial frontal gyrus, insula, limbic lobe, superior and middle temporal gyrus, and precentral gyrus), which have been associated with maternal networks during the postpartum period. Additionally, the changes in GM and WM volumes were related to changes in empathetic abilities in mothers. CONCLUSION: These results suggest that the brains of mothers exhibit adaptive structural dynamic plasticity. These findings provide a neuroanatomical basis for understanding how mothers process emotional sensory information during the postpartum period.

Structural changes of cerebellum and brainstem in migraine without aura.

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.

Seasonal and Gender Differences in Presence of Rickettsia felis and Blood meals Provide Additional Evidence of a Vector Role for Mosquitoes.

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 < 10-4) than in the months above 23°C. Collectively, the seasonal and gender differences of R. felis and blood meal in mosquitoes add to the existing evidence, supporting a potential vector role of mosquitoes in the transmission of R. felis. Studies with a R. felis infection model covering the full life cycle of mosquitoes is necessary to unambiguously prove the transstadial and transovarial transmission of R. felis in mosquitoes.

Molecular Detection of Bartonella spp. in China and St. Kitts.

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.

Housefly (Musca domestica) and Blow Fly (Protophormia terraenovae) as Vectors of Bacteria Carrying Colistin Resistance Genes.

Flies have the capacity to transfer pathogens between different environments, acting as one of the most important vectors of human diseases worldwide. In this study, we trapped flies on a university campus and tested them for mobile resistance genes against colistin, a last-resort antibiotic in human medicine for treating clinical infections caused by multidrug-resistant Gram-negative bacteria. Quantitative PCR assays we developed showed that 34.1% of Musca domestica (86/252) and 51.1% of Protophormia terraenovae (23/45) isolates were positive for the mcr-1 gene, 1.2% of M. domestica (3/252) and 2.2% of P. terraenovae (2.2%, 1/45) isolates were positive for mcr-2, and 5.2% of M. domestica (13/252) and 44.4% of P. terraenovae (20/45) isolates were positive for mcr-3 Overall, 4.8% (9/189) of bacteria isolated from the flies were positive for the mcr-1 gene (Escherichia coli: 8.3%, 4/48; Enterobacter cloacae: 12.5%, 1/8; Providencia alcalifaciens: 11.8%, 2/17; Providencia stuartii: 4.9%, 2/41), while none were positive for mcr-2 and mcr-3 Four mcr-1-positive isolates (two P. stuartii and two P. alcalifaciens) from blow flies trapped near a dumpster had a MIC for colistin above 4 mg/ml. This study reports mcr-1 carriage in Providencia spp. and detection of mcr-2 and mcr-3 after their initial identification in Belgium and China, respectively. This study suggests that flies might contribute significantly to the dissemination of bacteria, carrying these genes into a large variety of ecological niches. Further studies are warranted to explore the roles that flies might play in the spread of colistin resistance genes.IMPORTANCE Antimicrobial resistance is recognized as one of the most serious global threats to human health. An option for treatment of the Gram-negative ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) bacteria with multiple drug resistance was the reintroduction of the older antibiotic colistin. However, a mobile colistin resistance gene (mcr-1) has recently been found to occur widely; very recently, two other colistin resistance genes (mcr-2 and mcr-3) have been identified in Belgium and China, respectively. In this study, we report the presence of colistin resistance genes in flies. This study also reports the carriage of colistin resistance genes in the genus Providencia and detection of mcr-2 and mcr-3 after their initial identification. This study will stimulate more in-depth studies to fully elucidate the transmission mechanisms of the colistin resistance genes and their interaction.

BAALC and ERG expression levels at diagnosis have no prognosis impact on acute myeloid leukemia patients undergoing allogeneic hematopoietic stem cell transplantation.

Brain and acute leukemia, cytoplasmic (BAALC) and ETS-related gene (ERG) expression levels are independent prognostic factors for acute myeloid leukemia (AML); however, their prognostic impacts on AML patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) require further investigation. We studied 71 de novo AML patients treated with allo-HSCT and defined low and high expressers according to the median expression levels of BAALC and ERG at diagnosis respectively. High BAALC expression was associated with wild-type NPM1 (P = 0.000) and RUNX1 mutations (P = 0.027). High ERG expression was associated with FLT3-ITD absence (P = 0.003) and wild-type NPM1 (P = 0.001). BAALC and ERG expression levels were significantly correlated with each other (P = 0.001). Survival analyses including Kaplan-Meier curves and univariate and multivariate analysis consistently reported that there were no significant differences for both event-free survival (EFS) and overall survival (OS) (all P > 0.1), between high versus low BAALC and ERG expressers. Our study suggested that despite of their well-known adverse role in prognosis of AML, neither BAALC nor ERG expression levels at diagnosis had effect on survival of AML patients who underwent allo-HSCT.

Physiological and metabolic differences between visceral and subcutaneous adipose tissues in Nile tilapia (Oreochromis niloticus).

Visceral adipose tissue (VAT) and subcutaneous adipose tissue (SCAT) have different structures and metabolic functions and play different roles in the regulation of the mammal endocrine system. However, little is known about morphology and physiological and metabolic functions between VAT and SCAT in fish. We compared the morphological, physiological, and biochemical characteristics of VAT and SCAT in Nile tilapia and measured their functions in energy intake flux, lipolytic ability, and gene expression patterns. SCAT contained more large adipocytes and nonadipocytes than VAT in Nile tilapia. VAT had higher lipid content and was the primary site for lipid deposition. Conversely, SCAT had higher hormone-induced lipolytic activity. Furthermore, SCAT had a higher percentage of monounsaturated and lower polyunsaturated fatty acids than VAT. SCAT had higher mitochondrial DNA, gene expression for fatty acid ß-oxidation, adipogenesis, and brown adipose tissue characteristics, but it also had a lower gene expression for inflammation and adipocyte differentiation than VAT. SCAT and VAT have different morphological structures, as well as physiological and metabolic functions in fish. VAT is the preferable lipid deposition tissue, whereas SCAT exhibits higher lipid catabolic activity than VAT. The physiological functions of SCAT in fish are commonly overlooked. The present study indicates that SCAT has specific metabolic characteristics that differ from VAT. The differences between VAT and SCAT should be considered in future metabolism studies using fish as models, either in biomedical or aquaculture studies.