An operational robotic pollen monitoring network based on automatic image recognition.

There is high demand for online, real-time and high-quality pollen data. To the moment pollen monitoring has been done manually by highly specialized experts. Here we evaluate the electronic Pollen Information Network (ePIN) comprising 8 automatic BAA500 pollen monitors in Bavaria, Germany. Automatic BAA500 and manual Hirst-type pollen traps were run simultaneously at the same locations for one pollen season. Classifications by BAA500 were checked by experts in pollen identification, which is traditionally considered to be the "gold standard" for pollen monitoring. BAA500 had a multiclass accuracy of over 90%. Correct identification of any individual pollen taxa was always >85%, except for Populus (73%) and Alnus (64%). The BAA500 was more precise than the manual method, with less discrepancies between determinations by pairs of automatic pollen monitors than between pairs of humans. The BAA500 was online for 97% of the time. There was a significant correlation of 0.84 between airborne pollen concentrations from the BAA500 and Hirst-type pollen traps. Due to the lack of calibration samples it is unknown which instrument gives the true concentration. The automatic BAA500 network delivered pollen data rapidly (3 h delay with real-time), reliably and online. We consider the ability to retrospectively check the accuracy of the reported classification essential for any automatic system.

Assessing the Bone Age of Children in an Automatic Manner Newborn to 18 Years Range.

Bone age assessment (BAA) is a radiological process to identify the growth disorders in children. Although this is a frequent task for radiologists, it is cumbersome. The objective of this study is to assess the bone age of children from newborn to 18 years old in an automatic manner through computer vision methods including histogram of oriented gradients (HOG), local binary pattern (LBP), and scale invariant feature transform (SIFT). Here, 442 left-hand radiographs are applied from the University of Southern California (USC) hand atlas. In this experiment, for the first time, HOG-LBP-dense SIFT features with background subtraction are applied to assess the bone age of the subject group. For this purpose, features are extracted from the carpal and epiphyseal regions of interest (ROIs). The SVM and 5-fold cross-validation are used for classification. The accuracy of female radiographs is 73.88% and of the male is 68.63%. The mean absolute error is 0.5 years for both genders' radiographs. The accuracy a within 1-year range is 95.32% for female and 96.51% for male radiographs. The accuracy within a 2-year range is 100% and 99.41% for female and male radiographs, respectively. The Cohen's kappa statistical test reveals that this proposed approach, Cohen's kappa coefficients are 0.71 for female and 0.66 for male radiographs, p value < 0.05, is in substantial agreement with the bone age assessed by experienced radiologists within the USC dataset. This approach is robust and easy to implement, thus, qualified for computer-aided diagnosis (CAD). The reduced processing time and number of ROIs facilitate BAA.

Recombinant Adeno-Associated Viral Vectors (rAAV)-Vector Elements in Ocular Gene Therapy Clinical Trials and Transgene Expression and Bioactivity Assays.

Inherited retinal dystrophies and optic neuropathies cause chronic disabling loss of visual function. The development of recombinant adeno-associated viral vectors (rAAV) gene therapies in all disease fields have been promising, but the translation to the clinic has been slow. The safety and efficacy profiles of rAAV are linked to the dose of applied vectors. DNA changes in the rAAV gene cassette affect potency, the expression pattern (cell-specificity), and the production yield. Here, we present a library of rAAV vectors and elements that provide a workflow to design novel vectors. We first performed a meta-analysis on recombinant rAAV elements in clinical trials (2007-2020) for ocular gene therapies. We analyzed 33 unique rAAV gene cassettes used in 57 ocular clinical trials. The rAAV gene therapy vectors used six unique capsid variants, 16 different promoters, and six unique polyadenylation sequences. Further, we compiled a list of promoters, enhancers, and other sequences used in current rAAV gene cassettes in preclinical studies. Then, we give an update on pro-viral plasmid backbones used to produce the gene therapy vectors, inverted terminal repeats, production yield, and rAAV safety considerations. Finally, we assess rAAV transgene and bioactivity assays applied to cells or organoids in vitro, explants ex vivo, and clinical studies.

A novel agent attenuates cardiotoxicity and improves antitumor activity of doxorubicin in breast cancer cells.

Doxorubicin (Dox) is a well-known chemotherapeutic agent used in the treatment of various cancers. However, Dox-induced cardiotoxicity limits its further clinical use. We have previously reported a small molecular named biotin-conjugated ADTM analog (BAA) that exhibits cytoprotective effects against oxidative stress-induced cell injury in cardiomyoblast H9c2 cells. Here, the protective effects of BAA, indexed by attenuation of the cardiotoxicity induced by Dox as well as synergistic antitumor activity that increases the chemotherapeutic efficacy of Dox were investigated. Our results demonstrated that BAA significantly ameliorated Dox-induced toxicity in the H9c2 cells and zebrafish models. In addition, BAA attenuated Dox-induced endoplasmic reticulum (ER) stress in H9c2 cells. An ER stress inhibitor, 4-phenylbutyric acid, reversed the protective effect of BAA in H9c2 cells. In contrast, in human breast tumor MDA-MB-231 cells, BAA significantly enhanced Dox-induced cytotoxicity through upregulating Dox-induced ER stress response. Taken together, our findings indicate that Dox combined with BAA can significantly enhance its antitumor activity in breast cancer cells and reduce its cardiotoxicity, at least in part, by mediating ER stress activation.

Polycyclic aromatic hydrocarbon exposure, obesity and childhood asthma in an urban cohort.

BACKGROUND: Exposure to traffic-related air pollutants, including polycyclic aromatic hydrocarbons (PAHs) from traffic emissions and other combustion sources, and childhood obesity, have been implicated as risk factors for developing asthma. However, the interaction between these two on asthma among young urban children has not been studied previously. METHODS: Exposure to early childhood PAHs was measured by two week residential indoor monitoring at age 5-6 years in the Columbia Center for Children's Environmental Health birth cohort (n=311). Semivolatile [e.g., methylphenanthrenes] and nonvolatile [e.g., benzo(a)pyrene] PAHs were monitored. Obesity at age 5 was defined as a body mass index (BMI) greater than or equal to the 95th percentile of the year 2000 age- and sex-specific growth charts (Center for Disease Control). Current asthma and recent wheeze at ages 5 and 7 were determined by validated questionnaires. Data were analyzed using a modified Poisson regression in generalized estimating equations (GEE) to estimate relative risks (RR), after adjusting for potential covariates. RESULTS: Neither PAH concentrations or obesity had a main effect on asthma or recent wheeze. In models stratified by presence/absence of obesity, a significant positive association was observed between an interquartile range (IQR) increase in natural log-transformed 1-methylphenanthrene (RR [95% CI]: 2.62 [1.17-5.88] with IQRln=0.76), and 9-methylphenanthrene (2.92 [1.09-7.82] with IQRln=0.73) concentrations and asthma in obese children (n=63). No association in non-obese (n=248) children was observed at age 5 (Pinteraction<0.03). Similar associations were observed for 3-methylphenanthrene, 9-methylphenanthrene, and 3,6-dimethylphenanthrene at age 7. CONCLUSIONS: Obese young children may be more likely to develop asthma in association with greater exposure to PAHs, and methylphenanthrenes in particular, than non-obese children.

Neurodevelopmental toxicity of a ubiquitous disinfection by-product, bromoacetic acid, in Zebrafish (Danio rerio).

Disinfection of public drinking water and swimming pools is crucial for preventing waterborne diseases, but it can produce harmful disinfection by-products (DBPs), increasing the risk of various diseases for those frequently exposed to such environments. Bromoacetic acid (BAA) is a ubiquitous DBP, with toxicity studies primarily focused on its in vitro cytotoxicity, and insufficient research on its neurodevelopmental toxicity. Utilizing zebrafish as a model organism, this study comprehensively explored BAA's toxic effects and uncovered the molecular mechanisms through neurobehavioral analysis, in vivo two-photon imaging, transcriptomic sequencing, pharmacological intervention and molecular biological detection. Results demonstrated BAA induced significant changes on various indicators in the early development of zebrafish. Furthermore, BAA disrupted behavioral patterns in zebrafish larvae across locomotion activity, light-dark stimulation, and vibration stimulation paradigms. Subsequent investigation focused on larvae revealed BAA inhibited neuronal development, activated neuroinflammatory responses, and altered vascular morphology. Transcriptomic analysis revealed BAA-stressed zebrafish exhibited downregulation of visual transduction-related genes and activation of ferroptosis and cellular apoptosis. Neurobehavioral disorders were recovered by inhibiting ferroptosis and apoptosis. This study elucidates the neurodevelopmental toxicity associated with BAA, which is crucial for understanding health risks of DBPs and for the development of more effective detection methods and regulatory strategies.

Exposure to BaA inhibits trophoblast cell invasion and induces miscarriage by regulating the DEC1/ARHGAP5 axis and promoting ubiquitination-mediated degradation of MMP2.

Benz[a]anthracene (BaA), a hazardous polycyclic aromatic hydrocarbon classified by the EPA, is a probable reproductive toxicant. Epidemiological studies suggest that BaA exposure may be a risk factor for recurrent miscarriage (RM). However, the underlying mechanisms are not well understood. This study identified DEC1 as a key gene through RNA-seq and single-cell RNA sequencing analysis. DEC1 expression was found to be downregulated in villous tissues from women with RM and in primary extravillous trophoblasts (EVTs) exposed to BaA. BaA suppressed DEC1 expression by promoting abnormal methylation patterns. Further analysis revealed that ARHGAP5 is a direct target of DEC1 in EVTs, where DEC1 inhibits trophoblast invasion by directly regulating ARHGAP5 transcription. Additionally, BaA destabilized matrix metalloproteinase 2 (MMP2) by activating the aryl hydrocarbon receptor (AhR) and promoting E3 ubiquitin ligase MID1-mediated degradation. In a mouse model, BaA induced miscarriage by modulating the DEC1/ARHGAP5 and MID1/MMP2 axes. Notably, BaA-induced miscarriage in mice was prevented by DEC1 overexpression or MID1 knockdown. These findings indicate that BaA exposure leads to miscarriage by suppressing the DEC1/ARHGAP5 pathway and enhancing the MID1/MMP2 pathway in human EVTs.

Bone age assessment by multi-granularity and multi-attention feature encoding.

Background: Bone age assessment (BAA) is crucial for the diagnosis of growth disorders and the optimization of treatments. However, the random error caused by different observers' experiences and the low consistency of repeated assessments harms the quality of such assessments. Thus, automated assessment methods are needed. Methods: Previous research has sought to design localization modules in a strongly or weakly supervised fashion to aggregate part regions to better recognize subtle differences. Conversely, we sought to efficiently deliver information between multi-granularity regions for fine-grained feature learning and to directly model long-distance relationships for global understanding. The proposed method has been named the "Multi-Granularity and Multi-Attention Net (2M-Net)". Specifically, we first applied the jigsaw method to generate related tasks emphasizing regions with different granularities, and we then trained the model on these tasks using a hierarchical sharing mechanism. In effect, the training signals from the extra tasks created as an inductive bias, enabling 2M-Net to discover task relatedness without the need for annotations. Next, the self-attention mechanism acted as a plug-and-play module to effectively enhance the feature representation capabilities. Finally, multi-scale features were applied for prediction. Results: A public data set of 14,236 hand radiographs, provided by the Radiological Society of North America (RSNA), was used to develop and validate 2M-Net. In the public benchmark testing, the mean absolute error (MAE) between the bone age estimates of the model and of the reviewer was 3.98 months (3.89 months for males and 4.07 months for females). Conclusions: By using the jigsaw method to construct a multi-task learning strategy and inserting the self-attention module for efficient global modeling, we established 2M-Net, which is comparable to the previous best method in terms of performance.

Comparative genomics of Deinococcus radiodurans: unveiling genetic discrepancies between ATCC 13939K and BAA-816 strains.

The Deinococcus genus is renowned for its remarkable resilience against environmental stresses, including ionizing radiation, desiccation, and oxidative damage. This resilience is attributed to its sophisticated DNA repair mechanisms and robust defense systems, enabling it to recover from extensive damage and thrive under extreme conditions. Central to Deinococcus research, the D. radiodurans strains ATCC BAA-816 and ATCC 13939 facilitate extensive studies into this remarkably resilient genus. This study focused on delineating genetic discrepancies between these strains by sequencing our laboratory's ATCC 13939 specimen (ATCC 13939K) and juxtaposing it with ATCC BAA-816. We uncovered 436 DNA sequence differences within ATCC 13939K, including 100 single nucleotide variations, 278 insertions, and 58 deletions, which could induce frameshifts altering protein-coding genes. Gene annotation revisions accounting for gene fusions and the reconciliation of gene lengths uncovered novel protein-coding genes and refined the functional categorizations of established ones. Additionally, the analysis pointed out genome structural variations due to insertion sequence (IS) elements, underscoring the D. radiodurans genome's plasticity. Notably, ATCC 13939K exhibited a loss of six ISDra2 elements relative to BAA-816, restoring genes fragmented by ISDra2, such as those encoding for α/ß hydrolase and serine protease, and revealing new open reading frames, including genes imperative for acetoin decomposition. This comparative genomic study offers vital insights into the metabolic capabilities and resilience strategies of D. radiodurans.

Investigation of the photolysis process of benzo(a)anthracene (BaA) on polyvinyl chloride (PVC) and polystyrene (PS) microplastics: Plastics aging effect, transformation products and toxicity assessment.

Microplastics (MPs) and persistent pollutants (POPs) are new pollutants that are extensively studied worldwide. To fill the gaps that the degradation processes and mechanisms of polycyclic aromatic hydrocarbons (PAHs) on the surface of most MPs are still unclear, the photochemical transformation of benzo(a)anthracene (BaA) on polyvinyl chloride (PVC) MPs and polystyrene (PS) MPs in water were investigated and compared. The photolysis of BaA on the surface of PS in water proceeded easier than that on PVC within the 48 h irradiation period, with the pseudo-first-order rate constant of 0.0489 min-1 and 0.0181 min-1, respectively, which can be ascribed to the smaller particle size and more OH production of PS MPs. Due to the light competition between the chromophore and BaA as well as the light-shielding effect, aged MPs showed an inhibitory effect on the degradation of BaA compared with pristine MPs. For BaA/PVC MPs system, the degradation of BaA in real water was not significantly affected by coexisting ions and humic acid (HA) (p < 0.05), while slight inhibitory effect on the degradation of BaA appeared for PS MPs in different water matrices (UP: 86.97 %, YR: 84.47 %, PR: 81.42 % and HR: 83.21 %). According to the electron paramagnetic resonance (EPR) test, quenching experiment and probe experiment, the relative contribution of direct photolysis (PVC: 82.02 %; PS: 69.54 %) and indirect photolysis (PVC: 17.98 %; PS: 30.46 %) was confirmed. A total of 14 products were identified, and the product types were not affected by plastics aging. The results of the toxicity assessment indicated that although some intermediate products remained toxic to aquatic organisms, the toxicity of most products was lower than that of BaA. This study provides new insights into the environmental fate of PAHs and the role of MPs in the photolysis process of contaminants in surface water.

Effect of 16 pure hydrocarbons on the stabilization and lysis of fish (mudskipper: Boleophthalmus dussumieri) erythrocytes.

The in vitro effects of polycyclic aromatic hydrocarbons (PAHs) on erythrocyte membrane stability of the mudskipper (i.e., Boleophthalmus dussumieri) were tested by using field concentrations, acute and chronic potency divisor concentrations. This was achieved by studying their lytic or antilytic effects on fish erythrocytes in critical hypotonic saline media. The interaction of PAHs acute potency divisor concentrations with mudskipper erythrocyte causes dramatic changes in the structure of the membrane. A significant difference (p<0.05) was found between the control and treatment groups of mudskipper erythrocyte exposed to acute potency divisor concentrations. No significant difference (p>0.05) was observed between the control and the treatment groups of mudskipper erythrocyte exposed to field concentrations. The results showed that chronic potency divisor concentrations of PAHs protect mudskipper erythrocyte against osmotic hemolysis. Our results could be extended to the use of Erythrocyte Osmotic Fragility (EOF) test as a biochemical marker of membrane toxicity in marine pollution biomonitoring. However, results showed that membrane stability is not an appropriate biomarker for PAHs pollution after short exposure duration.

Whole-Blood Metabolomics of a Rat Model of Repetitive Concussion.

Mild traumatic brain injury (mTBI) and repetitive mTBI (RmTBI) are silent epidemics, and so far, there is no objective diagnosis. The severity of the injury is solely based on the Glasgow Coma Score (GCS) scale. Most patients suffer from one or more behavioral abnormalities, such as headache, amnesia, cognitive decline, disturbed sleep pattern, anxiety, depression, and vision abnormalities. Additionally, most neuroimaging modalities are insensitive to capture structural and functional alterations in the brain, leading to inefficient patient management. Metabolomics is one of the established omics technologies to identify metabolic alterations, mostly in biofluids. NMR-based metabolomics provides quantitative metabolic information with non-destructive and minimal sample preparation. We employed whole-blood NMR analysis to identify metabolic markers using a high-field NMR spectrometer (800 MHz). Our approach involves chemical-free sample pretreatment and minimal sample preparation to obtain a robust whole-blood metabolic profile from a rat model of concussion. A single head injury was given to the mTBI group, and three head injuries to the RmTBI group. We found significant alterations in blood metabolites in both mTBI and RmTBI groups compared with the control, such as alanine, branched amino acid (BAA), adenosine diphosphate/adenosine try phosphate (ADP/ATP), creatine, glucose, pyruvate, and glycerphosphocholine (GPC). Choline was significantly altered only in the mTBI group and formate in the RmTBI group compared with the control. These metabolites corroborate previous findings in clinical and preclinical cohorts. Comprehensive whole-blood metabolomics can provide a robust metabolic marker for more accurate diagnosis and treatment intervention for a disease population.

SMANet: multi-region ensemble of convolutional neural network model for skeletal maturity assessment.

Background: Bone age assessment (BAA) is a crucial research topic in pediatric radiology. Interest in the development of automated methods for BAA is increasing. The current BAA algorithms based on deep learning have displayed the following deficiencies: (I) most methods involve end-to-end prediction, lacking integration with clinically interpretable methods; (II) BAA methods exhibit racial and geographical differences. Methods: A novel, automatic skeletal maturity assessment (SMA) method with clinically interpretable methods was proposed based on a multi-region ensemble of convolutional neural networks (CNNs). This method predicted skeletal maturity scores and thus assessed bone age by utilizing left-hand radiographs and key regional patches of clinical concern. Results: Experiments included 4,861 left-hand radiographs from the database of Beijing Jishuitan Hospital and revealed that the mean absolute error (MAE) was 31.4±0.19 points (skeletal maturity scores) and 0.45±0.13 years (bone age) for the carpal bones-series and 29.9±0.21 points and 0.43±0.17 years, respectively, for the radius, ulna, and short (RUS) bones series based on the Tanner-Whitehouse 3 (TW3) method. Conclusions: The proposed automatic SMA method, which was without racial and geographical influence, is a novel, automatic method for assessing childhood bone development by utilizing skeletal maturity. Furthermore, it provides a comparable performance to endocrinologists, with greater stability and efficiency.

Brachiocephalic and Subclavian Artery Aneurysm Treated With Covered Stent.

Endovascular repair for brachiocephalic artery aneurysms (BAAs) and subclavian artery aneurysms (SAAs) is often unclear. We treated 2 such cases percutaneously with covered stents. It may be an alternative to surgical repair for patients with isolated BAA or SAA. (Level of Difficulty: Advanced.).

Deep longitudinal phenotyping of wearable sensor data reveals independent markers of longevity, stress, and resilience.

Biological age acceleration (BAA) models based on blood tests or DNA methylation emerge as a de facto standard for quantitative characterizations of the aging process. We demonstrate that deep neural networks trained to predict morbidity risk from wearable sensor data can provide a high-quality and cheap alternative for BAA determination. The GeroSense BAA model was trained and validated using steps per minute recordings from 103,830 one-week long and 2,599 of up to 2 years-long longitudinal samples and exhibited a superior association with life-expectancy over the average number of steps per day in, e.g., groups stratified by professional occupations. The association between the BAA and effects of lifestyles, the prevalence of future incidence of diseases was comparable to that of BAA from models based on blood test results. Wearable sensors let sampling of BAA fluctuations at time scales corresponding to days and weeks and revealed the divergence of organism state recovery time (resilience) as a function of chronological age. The number of individuals suffering from the lack of resilience increased exponentially with age at a rate compatible with Gompertz mortality law. We speculate that due to the stochastic character of BAA fluctuations, its mean and auto-correlation properties together comprise the minimum set of biomarkers of aging in humans.

Implementation of teledentistry for orthodontic practices.

Recent advances in technology, growing patient demand, and the need for social distancing due to Coronavirus Disease 2019 has expedited adoption of teledentistry in orthodontics as a means of consulting and monitoring a patient without an in-office visit. However, a lack of computer literacy and knowledge of software choices, and concerns regarding patient safety and potential infringement of regulations can make venturing into this new technology intimidating. In this article, various types of teledentistry systems for orthodontic practices, implementation guidelines, and important regulatory considerations on the use of teledentistry for orthodontic purposes are discussed. A thorough evaluation of the intended use of the software should precede commitment to a service. Selected service should be Health Insurance Portability and Accountability Act compliant at minimum and a Business Associate Agreement should be in place for protection of privacy. Ensuring the compatibility of the designated clinic computer with the system's requirements and installation of all safeguards must follow. Appointments should be documented in the same manner as in-office visits and teledentistry patients must be located within the clinician's statutory license boundary. Informed consent forms should include teledentistry or a supplemental teledentistry consent form should be used. Malpractice insurance covers everything usual and customary under the provider's license but the need for cyber liability insurance increases with teledentistry.

Highly self-healable and injectable cellulose hydrogels via rapid hydrazone linkage for drug delivery and 3D cell culture.

To meet the rising demand of injectable hydrogels with self-healing, robustness and biocompatibility for biomedical engineering, the reversible ketoester-type acylhydrazone linkages was used for the fabrication of novel cellulose-based hydrogel. The ketoester-type acylhydrazone bond exchanged rapidly, endowing the hydrogels with highly efficient self-healing performance without any external stimuli under physiological environment, which was hardly achieved with the widely used arylhydrozone bond. The dynamic hydrogels exhibited tunable mechanical property, pH responsiveness, injectability and biocompatibility, demonstrating immense applications prospect for various biomedicines, such as drug and cell delivery. The pH-responsive controlled release of model drug doxorubicin (DOX) loaded in the hydrogel was demonstrated. In addition, benefitting from the excellent biocompatibility and the reversible ketoester-type acylhydrazone bonds, cells were encapsulated in the hydrogels as 3D carrier. The covalent adaptable network intensified injectability of cell-laden hydrogels and improved the long-lasting viability for cell culture, showing great potential in the biomedical field.

Different unfolding pathways of homologous alpha amylases from Bacillus licheniformis (BLA) and Bacillus amyloliquefaciens (BAA) in GdmCl and urea.

Understanding the factors governing stability of proteins is fundamentally and industrially important topic in protein science. Bacterial alpha amylases are industrially important enzymes which are involved in the breakage of α-1, 4-glycosidic bonds in starch. Current study is focussed on elucidating the role of non-covalent interactions in the differential stability of alpha amylases from thermophilic like Bacillus licheniformis (BLA) and mesophilic Bacillus amyloliquefaciens (BAA). The conformational stability of BLA is slightly higher than BAA in GdmCl which are 2.94 and 2.53 kcal/mol respectively. BLA does not unfold even in 8.0 M urea at pH 7.0, while for BAA, the conformational stability in urea is calculated to be 2.22 kcal/mol. A structure-function relationship study of BLA reveals the non-coincidental unfolding by far UV-CD, enzyme activity and tryptophan fluorescence which indicates the presence of partially unfolded intermediates. The existence of intermediates in BLA during GdmCl induced unfolding was further confirmed by ANS fluorescence. The unfolding kinetics of both enzymes showed biphasic nature with slower unfolding of BLA compare to BAA pointing towards the higher kinetic stability of BLA than BAA. Taken together, our work demonstrates that the higher stability of BLA is mainly due to the combination of ionic and hydrophobic interactions.

Bulleyaconitine A Exerts Antianxiety and Antivisceral Hypersensitivity Effects.

Visceral pain is one of the leading causes for abdominal pain in gastroenterological diseases and is still hard to treat effectively. Bulleyaconitine A (BAA) is an aconitine analog and has been used for the treatment of pain. Our previous work suggested that BAA exerted analgesic effects on neuropathic pain through stimulating the expression of dynorphin A in spinal microglia. Here, we investigated the inhibitory effect of BAA on visceral pain and examined whether the expression of dynorphin A in spinal microglia was responsible for its effects. We found that BAA produced significant antivisceral pain effect induced by acetic acid through stimulating dynorphin A expression in spinal microglia. In addition, anxiety and chronic visceral pain are highly prevalent comorbid conditions in clinical research, which is still a problem to be solved. We also aimed to evaluate the effects of BAA on anxiety. A comorbidity model with characteristics of both chronic visceral pain and anxiety was developed by colorectal injection of 2,4,6-trinitrobenzene sulfonic acid and the induction of heterotypic intermittent chronic stress protocol. In comorbid animals, BAA exerted great antianxiety effects. Meanwhile, the antianxiety mechanism of BAA was different with the antivisceral pain mechanism of BAA. In conclusion, our study demonstrated, for the first time, that BAA exerted marked antivisceral pain and antianxiety effects, which expands the analgesic spectrum and clinical application of BAA. Furthermore, it also it provides a better guidance for the clinical use of BAA.

Substrate specificity of the galactokinase from the human gut symbiont Akkermansia muciniphila ATCC BAA-835.

Galactokinases, which catalyze the phosphorylation of galactose and possible other monosaccharides, can provide an activated sugar donor to synthesize sugar-containing molecules. In this study, a novel galactokinase from human gut symbiont Akkermansia muciniphila ATCC BAA-835 (GalKAmu) was expressed and characterized. GalKAmu displayed broad substrate tolerance, with catalytic activity towards Gal (100 %), GalN (100 %), GalA (20.2 %), Glc (52.5 %), GlcNAc (15.5 %), Xyl (<5%), ManNAc (58 %), ManF (37.4 %) and l-Glc (80 %). Most interestingly, this was the first GalK isoform which can tolerate ManNAc. Thus, our characterization of GalKAmu broadens the substrate selection of galactokinases.