Conventional invasive diagnostic imaging techniques do not adequately resolve complex Type B and C coronary lesions, which present unique challenges, require personalized treatment and result in worsened patient outcomes. These lesions are often excluded from large-scale non-invasive clinical trials and there does not exist a validated approach to characterize hemodynamic quantities and guide percutaneous intervention for such lesions. This work identifies key biomarkers that differentiate complex Type B and C lesions from simple Type A lesions by introducing and validating a coronary angiography-based computational fluid dynamic (CFD-CA) framework for intracoronary assessment in complex lesions at ultrahigh resolution. Among 14 patients selected in this study, 7 patients with Type B and C lesions were included in the complex lesion group including ostial, bifurcation, serial lesions and lesion where flow was supplied by collateral bed. Simple lesion group included 7 patients with lesions that were discrete,  Owing to their anatomic, physiological and functional difficulties, conventional diagnostic metrics are inadequate for assessing disease progression in complex lesions4. Secondary hemodynamic biomarkers are promising metrics for predicting outcome of complex lesions and thus guiding future treatment decisions. Thus, there is an urgent need to determine intracoronary hemodynamic quantities that can differentiate complex coronary lesions on a per-patient basis. Non-invasive computational diagnostic approaches can be particularly useful in assessing complex lesions. However, despite their usefulness in simple, single coronary lesions, the utility of these methods to diagnose complex coronary lesions remains elusive as these patients have been excluded in most clinical trials using such computational models6. In this study, we present a computational fluid dynamic (CFD) framework that accurately determines coronary arterial tree physiology at ultra-high resolutions. This method enables quantitative evaluation of intra-arterial hemodynamics in complex coronary diseases, such as ostial lesion, bifurcation lesion and serial lesion, with high accuracy and prediction reliability for the first time. The accuracy of our framework is determined by comparing derived diagnostic metrics to clinical measurements, such as fractional flow reserve (FFR) in both simple and complex coronary lesions.Complex coronary lesions, such as bifurcation and ostial lesions, are found in more than 20% of patients suffering from coronary diseases who are at high risk of developing secondary major adverse cardiac events4. A similar story prevails for ostial lesions which respond poorly to conventional interventional procedures and present higher procedural and medium-term complications8. Due to the morphological complexity of these lesions, data on the intracoronary hemodynamic characteristics in complex lesions remains unavailable8. This study addresses this limitation by proposing a CFD framework that naturally incorporates realistic patient anatomy and derives FFR and the local flow quantities, such as velocity and shear stress. FFR is the trans-stenotic pressure ratio measured to determine whether an arterial blockage should be stented. To supplement FFR, accurate biomechanical models can be used to establish a clinically-relevant metric to diagnose and predict disease progression.While it is known that using conventional diagnostic metrics to guide revascularization planning can reduce the need for unnecessary intervention in single main branch lesions, such benefits for complex coronary lesions lack compelling clinical evidence. Further, complications such as in-stent restenosis (ISR) occur in 30% of the bifurcation lesion cases following intervention9. CA offers high spatial and temporal resolution, superior to CT angiography, and can therefore can be used to incorporate the anatomical detail needed for complex arterial anatomy10.Complex lesions are challenging to model with existing computational frameworks due to anatomic and functional difficulties which require (1) a high-resolution imaging modality and (2) physiological parameters that can lead to instabilities and inaccuracies in numerical approaches. Therefore, to fully elucidate the anatomy-function relationship of arterial hemodynamics in complex lesions, our computational framework comprises two steps: first, reconstructing complete arterial trees using coronary angiography (CA) imaging data and, second, performing hemodynamic simulations using HARVEY (v2) - a massively parallel CFD solver. Using 3D geometries derived from CA data with secondary and tertiary bifurcations is a key development and has required integration of a biplane reconstruction algorithm11. Using parallel computing, this framework enables ultra-high resolution physiological modeling in reasonable execution times, thus meeting the high computational demand needed for these in silico experiments. Fast and accurate non-invasive functional arterial assessment is achieved by using the massively parallel CFD solver HARVEY (v2)13. With this CFD-CA framework, we expect to non-invasively derive diagnostic metrics, such as FFR with lower errors and reclassification rates, and resolve local hemodynamics for pathophysiological analysis in all identifiable vessels in an angiogram. We believe such hemodynamic profiling at the clinical level will be especially useful in complex coronary lesion which continue to suffer from high rate of surgical complications and major adverse cardiac events8.Our CFD framework allows physiological maps to be overlaid on conventional imaging data which can be directly used to determine hemodynamic differences across the entire arterial tree. The size of the smallest vessels that can be modeled imposes restrictions on maximum achievable resolution due to the large computational load15. To validate the reconstructed geometries, an experienced interventional cardiologist compared the anatomy of the reconstructed arterial models to the original patient angiogram used as input to the reconstruction algorithm. The arterial architecture was compared in terms of number of vessels, vessel type , branching patterns (bifurcation and trifurcation) and stenoses anatomy . A quantitative comparison was established by aligning the reconstructed models to match arterial orientation in the angiograms and measuring the ostium diameter and minimal luminal diameter of the lesion using ImageJ (v1.52k)16. The numerical results of these measurements can be found in Supplementary Table 12. LBM has been extensively used to efficiently model hemodynamics in patient-specific geometries22. More recent studies have leveraged LBM-based CFD solvers to derive pressure fields required to compute FFR24. In contrast to other CFD solvers, LBM-based solvers allow efficient distribution of computational load and high-resolution simulations on millions of processors17. HARVEY (v2) used the 3D coronary arterial mesh in standard stereolithography (STL) format as input to the model.This study uses 14 patient datasets acquired from Brigham and Women’s Hospital which had 2D angiograms and invasively measured FFR values at the time of coronary angiography  and coronary dominance measures for each patient under resting or basal flow conditions  and the dependent variable is measured with error. For the resting gradient data we computed an intercept of Bland Altman analysis further confirmed the agreement between CFD-CA and invasive measures. Figure 33. The computed CFD-CA FFR was also categorized in the same way and assessed for comparison with invasive FFR  local hemodynamics differences exist between complex and non-complex lesions included in this study, and (2) differences in local hemodynamics are exacerbated for complex lesion cases which could help understand worsened outcomes in patients with coronary artery disease.37. Therefore, supplementing the knowledge of FFR with a rich 3D hemodynamic environment, by investigating spatial flow heterogeneity under resting and hyperemic conditions, can help in prospectively evaluating frequently witnessed adverse events related to complex lesions.We test these hypothesis for the specific lesions included in this study. Our rationale for these hypotheses was that previous studies have shown that arterial shear stress,  1.5 Pa, is considered atheroprotective and >2.5 Pa can lead to pro-atherosclerotic plaque rupture38. We tested our hypotheses by analyzing two characteristic hemodynamic risk-factors: endothelial shear stress (ESS) and velocity at resting and hyperemic states40. ESS was averaged over the period of cardiac cycle and calculated as time-averaged endothelial shear stress (TAESS) and velocity was computed as the circumferential average velocity. Both quantities were then calculated along the entire lesion length. Figures We note resting gradient and FFR values between complex and non-complex lesions do not necessarily differ, and local physiological changes that arise directly due to the complicated anatomy of complex lesions could better help understand disease progression Fig. . This phTo test our hypotheses, we compared ESS and velocity for patients in complex and non-complex lesion groups. Figure p = 0.03] and 0.73 ± 0.18 [p = 0.09] respectively. And at hyperemic conditions, the ratio for ESS and velocity was 0.62 ± 0.2 [p = 0.03] and 0.7 ± 0.21 [p = 0.08] respectively.To determine the statistical significance of these hemodynamic differences we used a paired ratio t-test. Pairing between complex and non-complex case was done based on the inlet boundary condition. Specifically, a complex and non-complex case with similar average velocity flow at the ostium was paired. Averaged velocity at the ostium was computed by dividing the cardiac output, determined based on the coronary dominance, and ostium area. The cardiac output, coronary dominance and ostial diameter were measured by an experienced interventional cardiologist. This pairing allowed us to assess the hemodynamic differences arising due to the unique morphology of complex lesions compared to simple single, lesions, despite the similar inlet flow rate. For resting condition, the ratio of complex and non-complex lesions for ESS and velocity was 0.69 ± 0.1 [41, we believe these findings can be clinically important and used to guide treatment in patients with complex lesions. However, it is also worth noting that even simple, single lesions with high severity or flow rates could also lead to high ESS and velocity. And as such, these findings could be dependent on the choice of cases. Therefore, to provide a general evidence for ESS and velocity differences between lesion types the range of complex lesion cases should be representative of most anatomic variants and severity of different complex cases, and likewise for simple lesion cases. Overall such an analysis requires a large clinical study.For the specific cases included in this study, the ESS and velocity differences do not arise based just on the stenosis severity, but in fact due to the unique anatomy of complex lesions, since the percentage of stenosis in some complex lesion cases was less severe than the corresponding simple lesion case. In summary we note that for the particular cases in this study, ESS was significantly different between complex and non-complex lesion at both resting and hyperemic conditions and marginal significance was observed for velocity. As high ESS has been shown to predict adverse cardiac events, such as myocardial infarction42. As anticipated, CFD-CA FFR values were sensitive to anatomic variations attributed to percent stenoses, and FFR variation as high as 12% was noted for the 30% increase in percent stenoses  FFR predictions remained relatively unaltered, <2%, with variations in these clinical parameters for a given patient geometry Fig. . We thenses Fig. . Similar25. Thus several groups have attempted to apply computational models to medical imaging data to derive FFR with clinical accuracy, but continue to exclude complex lesions46. Furthermore, imaging techniques such as CT angiography, optical coherence tomography and rotational coronary angiography are neither universally available nor routinely performed in clinical practice, which limits the application of computational models that rely on these imaging datasets46. This study presents ultra-high resolution personalized CFD simulations to compute conventional diagnostic metrics and intra-arterial physiology in complex and non-complex coronary lesion using widely available coronary angiography imaging data.Recent years have witnessed dramatic increase in computational approaches for diagnosing cardiovascular diseases, which continue to dominate healthcare costs and are projected to be over 1 trillion dollars by 2035, according to the American Heart Association. Treatment of complex coronary lesions is involved in more than 20% of the total number of PCI procedures performed and post-interventional complications such as in-stent restenosis occurs in 30% of the cases. PCI challenges for complex lesion are further compounded by the limited accessibility and the high procedural and operational costs of traditional diagnostic methods, which have restricted its wide scale utilization and paved the way for computational methods that can non-invasively derive FFR in routine clinical practice49. This study interrogates ESS as a potential biomarker and builds on the current understanding in literature, while advancing it in a critical way. Capturing coronary physiology described by local hemodynamics throughout the arterial tree is crucial in order to examine the consequences of impaired coronary flow and plaque development. Physiological assessment of the arterial tree can enable an accurate and non-invasive approach in the diagnosis and treatment of coronary artery disease. Patient-specific characterization of local hemodynamic risk factors which have been previously shown to affect formation and progression of atherosclerosis, such as ESS and velocity, can account for complications in certain patient and complex lesion types that otherwise cannot be explained by ubiquitous procedural factors40. Such characterization is important because local hemodynamic metrics hold the potential to be clinically invaluable to physicians as traditional diagnostic metrics, such as FFR, do not differ between complex lesion anatomies relative to Type A lesions which have simpler morphology. Thus, by providing information about known hemodynamic risk factors, our CFD-CA methodology can aid clinical decision-making. In particular, spatial heterogeneity of local hemodynamic quantities holds enormous clinical relevance as it could provide grounds for prophylactic interventions and potentially avert any future adverse cardiac events, such as myocardial infarction41.The major findings of this study are: (1) traditional pressure-based metrics, such as FFR, do not significantly differ between non-complex Type A coronary lesions compared to complex lesions, (2) intra-arterial hemodynamics, which were previously unavailable, for complex coronary lesion can be accurately derived from our CFD-CA method, and (3) ESS and intra-lesion velocity are important biomarkers which significantly differ between complex and non-complex lesions and can help understand the reason for such patients developing secondary adverse cardiac events. These findings are critical as high ESS is predictive of clinical outcomes such as myocardial infarction and often alters atherosclerotic plaque characteristics10. For example compared to CTA, CA has spatial resolution of 150-200 mum vs 300-400 mum and temporal resolution of 10ms vs 80-190ms10. Therefore, deriving 3D arterial geometries based on CA renders itself to the capability of reconstructing complex anatomic regions and including greater number of side branches11. Accurate anatomy has a direct impact on the fidelity of CFD simulations, thus by relying on CA data, we were able to simulate flow in complex coronary lesions. We achieve this by using a validated 3D reconstruction algorithm as described in9 that uses pair of 2D angiograms and creates the 3D coronary skeleton by detecting vessel centerlines and cross-sectional diameter. This reconstruction algorithm enables capturing anatomic details of third and fourth order branching patterns and vessel tortuousity up to 1mm in diameter. However, a closely related to simulating flow in these complex 3D arterial geometries is that it poses formidable computational load. To this end, we coupled our CA reconstructions with a massively parallel CFD solver that efficiently scales on millions of processors and can accurately simulate flow in different arterial vasculature50. HARVEY (v2) has previously been validated with in vitro experiments in patient-derived 3D vascular geometries18. Despite the complexity and anatomic detail of arterial models, these geometries could be used without the need for mesh refinement on exterior surfaces because HARVEY (v2) is nearly agnostic to mesh triangulation. Such features enable integration of flow from smaller side branches to the main branch flow, creating realistic arterial tree physiology. Therefore, we methodologically advance existing state-of-art studies, which non-invasively compute diagnostic metrics using angiograms for 3D arterial reconstruction.The capability of our method to assess complex lesions comes from combining high-resolution coronary angiographic imaging data and a massively parallel hemodynamic simulator. CA has higher spatial and temporal resolution compared to other commonly used imaging modalities in 3D coronary flow models45. Such single vessel arterial models preclude the inclusion of collateral vessel bed and bifurcation lesions. Other angiogram-based studies, which incorporate an arterial tree up to first or second order vessel branching, have used a reduced-order lumped parameter model for computational modeling51. In reduced order models, the fluid dynamic equations are not solved, and instead the arterial network is treated as an electrical circuit and the vessel segments as resistors, which lowers the computational complexity52. With reduced order models, local intra-arterial physiology, WSS and velocity fields, cannot be obtained. Therefore, we note that existing CA-based computational studies either rely on a simplified 3D geometry or computational algorithm. Such simplifications potentially limit any complex lesion and local hemodynamic analysis for arterial networks and quantities, for example ESS and velocity, cannot be derived53. Therefore, to the best of our knowledge, this is the first CA-based 3D CFD method to simulate hyperemia and compute physiological flow parameters in complex and simple, single coronary lesions.Previous studies that have used routine angiographic data for non-invasive physiological assessment of coronary arteries typically use a simplified geometric model incorporating only the main vessel-of-interest with branching up to a single side vessel51. We establish the robustness of our CFD-CA methodology by performing sensitivity analysis and report near independence of computed FFR with respect to variability in clinical measurements, such as cardiac output, hematocrit and heart rate  and FFR  for both simple and complex lesion groups. These results hold equivalent-to-better limits of agreement with other current state-of-the-art computational FFR modelsTraditional diagnostic metrics such as resting gradient and FFR do not significantly differ between complex and non-complex lesions. Therefore, we investigated intra-arterial hemodynamics differences, such as ESS and velocity, which can help understand the reason for worsened clinical outcome of patients with complex lesions such as bifurcation, ostial and serial lesions.The physiological information, in the form of pressure, ESS and velocity for the complete arterial tree in primary, secondary and tertiary branches, is not clinically available to physicians. This analysis can be utilized to study prospective progression of atherosclerosis in arterial segments secondary to the main diseased vessels.By deriving the local hemodynamic physiology, we were able to study for the first time previously intractable relationship between complex and non-complex coronary lesions for pressure-based metrics, such as FFR and intra-arterial hemodynamics on a per-patient basis. This investigation is significant for two reasons:34. Second limitation is the small number of patient cases, and the high diagnostic accuracy reported for the 14 patient datasets may be serendipitous. Furthermore, the complex lesion also excluded vulnerable plaques because CA cannot easily detect calcification54. However, despite of excluding vulnerable plaques and the small cohort size, the patient population in this study represents a different complex lesion types and the boundary conditions were uniquely tuned to each patient case. Thus, the results reported here are encouraging and hypothesis-generating, and warrant study in a larger patient cohort with outcome data.There are several limitation in this study. First, wall motion is not considered this study. However, rigid walls is a common assumption used in computational fluid dynamic studies evaluating coronary hemodynamics and FFR, as it is not highly sensitive to hemodynamic conditions such as heart rate, blood pressure, and myocardial contractility57, another limitation of this work is that our CFD-CA FFR validation using invasive measurements does not directly establish the accuracy of velocity and ESS. However, in our previous works we validated HARVEY simulation using 3D printing and experimental results for physiologically relevant flows in vascular geometries50. In these works, complete velocity profile was validated with particle image velocimetry (PIV) data using physiological blood flow profiles and 3D printed aorta and femoral geometries, which differ in the type of flow and physical scale. As for the ESS calculation, the strain rate tensor 58. Therefore, the computational results are currently obtained offline, however, the use of massively parallel CFD solver enables near real-time ( 30 minutes) analysis.While effectiveness and robustness of the LBM for computing ESS in vascular regimes is well-establishedThe long-term goal of this study is to improve patient outcome by developing patient-specific multiscale CFD models based on conventional CA data. This approach is significant as the CFD-CA methodology enables seamless integration with routine cardiac catheterization laboratories by relying on conventional angiogram data. The proposed CFD technology solves fluid dynamic equations on every point of the 3D arterial geometry for patient cases displaying any degree of anatomic lesion complexity. Such unique features provide a complete road map of coronary physiology which describes the lesion-specific vessel and downstream effects on other vessels in the arterial tree. Our framework has minimal manual interaction for setting parameters and negligible variance with respect to uncertainties in model inputs. This invariance provides high prediction reliability of key hemodynamic risk factors and diagnostic metrics which is crucial for wide scale deployment in clinical decision-making.This study does not involve any experiments on humans and/or the use of human tissue samples. This study was approved by the Partners Human Research Committee Institutional Review Board who waived informed consent for the study  (v2) is based on the LBM, an alternate approach to solving the Navier-Stokes equations governing the fluid flow. Derived from the Boltzmann equation of kinetic theory, LBM describes the fluid by a probability distribution function x according to discrete velocities p and velocity u, are computed locally as moments of 61:61. The HARVEY (v2) implementation uses the standard three dimensional D3Q19 velocity discretization and the single relaxation time BGK collision kernel. The code is written in C and C++, using OpenMP and MPI for parallelization. Wall shear stress is evaluated at fluid points adjacent to the wall. The wall shear stress vector 57 :57. The pressure in lattice units was converted into physical units (mmHg) by using the pressure coefficient equation:u are the reference density and velocity, respectively. Further details about the numerical implementation, parallelization, and scaling of HARVEY (v2) can be found in17.CFD simulations were performed using HARVEY (v2), a computational hemodynamics software package26.To model the coronary circulation, we simulated blood as an incompressible Newtonian fluid with density of 1060 kg/m62. The Poiseuille equation is given as28:R is resistance of blood with viscosity 28. Vessel radius has a stronger effect on the overall flow compared to vessel length and viscosity, such that small changes in radius result in dramatic changes in vessel resistance and overall volume. This equation provides an important description for relating flow with vessel anatomy. With the assumption of Newtonian flow, it can be used for modeling physiological  and pathological  changes based on vessel radius and how that affect pressure and flow volume28.It is important to set up a CFD simulation individualized for each patient case by incorporating: cardiac output, heart rate, coronary dominance, blood pressure and vessel sizes. For the inlet boundary condition, the transient flow was considered using velocity waveform tuned to the patient parameters. Transient (or pulsatile) simulations are more realistic than steady simulations as they capture the periodically changing velocity during the cardiac cycle. Pulsatile ESS and pressure derived from transient simulations were thus varying in magnitude, unidirectional and averaged over the period of the cardiac cycle. A Poiseuille profile was prescribed at the inlet and finite difference boundary conditions were imposed63. The resistance based boundary conditions assume linear relationship between flow rate and pressure, but can be uniquely prescribed to each vessel outlet of the arterial network. Such an implementation for coronary circulation, while computationally more expensive than constant pressure boundary conditions, is needed to ensure the physiological behavior of arterial network as each arterial outlet offers different flow resistance and ESS due to different radii of arterial outlets63. To prevent noise, we averaged the diameter for each arterial outlet up to a length of 1.5mm from the vessel endings. Thus, for all patient cases, the coronary arterial microcirculation resistance (R) for each outlet i was computed based on the vessel diameter, mean flow (Q) and mean aortic pressure (P) using the following equation63:mum as the reference for spatial convergence and 5 cardiac cycles  and CFD-CA FFR (dependent variable). Continuous variables, such as the ESS and velocity, were described by mean and standard deviation. A two-tailed paired ratio t-test was used to assess whether mean differences between local hemodynamic quantities (ESS and velocity) were significant. A value of Supplementary Information


Gluconobacter is a genus of acetic acid bacteria (AAB) whose members have been shown to function as insect symbionts. Here, we report the complete genome sequence of Gluconobacter cerinus, isolated from field-collected Drosophila suzukii using a hybrid assembly approach. The data provide essential insights into the metabolic functions of the symbiont to the host. Gluconobacter is a genus of acetic acid bacteria (AAB) whose members have been shown to function as insect symbionts. Here, we report the complete genome sequence of Gluconobacter cerinus, isolated from field-collected Drosophila suzukii using a hybrid assembly approach. The data provide essential insights into the metabolic functions of the symbiont to the host. Gluconobacter (family Acetobacteraceae) is a member of the acetic acid bacteria (AAB) group. Recent research has highlighted AAB as widespread symbionts in insects, particularly insects of the orders Diptera, Hymenoptera, and Hemiptera  and shown to support host egg production  platforms.The bacterial genus emiptera . For exaoduction . RecentlD. suzukii were dissected and homogenized in phosphate-buffered saline (PBS). The homogenate was plated onto De Man, Rogosa and Sharpe (MRS) agar and incubated at 37°C for 48 h. Bacterial colonies were picked and grown in MRS broth overnight before being pelleted by centrifugation. Total genomic DNA was isolated using a phenol-chloroform protocol described previously  under the accession numbers SRR13680735 (ONT) and SRR13680736 (Illumina).This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under the accession number


Diet-induced obesity (DIO) reduces the orosensory perception of lipids in rodents and in some humans. Although bariatric surgery partially corrects this alteration, underlying mechanisms remain poorly understood. To explore whether metabolic changes might explain this fat taste disturbance, plasma metabolome analyses, two-bottle choice tests and fungiform papillae (Fun) counting were performed in vertical sleeve gastrectomized (VSG) mice and sham-operated controls. An exploratory clinic study was also carried out in adult patients undergone a VSG. In mice, we found that (i) the VSG reduces both the plasma neurotoxic signature due to the tryptophan/kynurenine (Trp/Kyn) pathway overactivation and the failure of fat preference found in sham-operated DIO mice, (ii) the activity of Trp/Kyn pathway is negatively correlated to the density of Fun, and (iii) the pharmacological inhibition of the Kyn synthesis mimics in non-operated DIO mice the positive effects of VSG . In humans, a reduction of the plasma Kyn level is only found in patients displaying a post-surgery improvement of their fat taste sensitivity. Altogether these data provide a plausible metabolic explanation to the degradation of the orosensory lipid perception observed in obesity. Recent studies suggest that the sense of taste might be compromised in obesity contributing to unhealthy food choices . By provBariatric surgeries are efficient procedures used to induce long-term weight loss and to improve associated comorbidities frequently found in obese patients . VerticaCollectively these data show that the obesity state plays a role in the efficiency of the orosensory perception of lipids and raise a basic question: how do obesity and bariatric surgeries interfere with the fatty taste detection? To explore this issue without preconception, a global analysis of plasma metabolites was performed in combination with behavioral preference tests and Fun counting in both DIO mice after VSG or sham-operation (Sham). A human exploratory study was also undertaken two weeks and six months after a vertical sleeve gastrectomy.These studies were conducted in the strict accordance with the European guidelines for the care and use of laboratory animals and the protocols were approved by the French National Animal Ethics Committee (APAFIS#12728-201712151028893 and APAFIS#19877-2019032109244847). Six-week-old C57Bl/6 male mice were purchased from Charles River Laboratories (France). They were individually housed in a controlled environment  and allowed free access to tap water and chow. Mice were fed either a standard chow  or an obesogenic high fat diet   for two n = 11) and in lean and obese controls after 9 weeks under HFD . After surgery, DIO mice were maintained on HFD until sacrifice.Firstly, to explore the molecular mechanisms by which bariatric surgery might affect the fat taste, a non a priori investigation  was undertaken after VSG in obese mice  by addition of 1-methyl tryptophan  in the drinking water according to [n = 15).Secondly, to determine the putative role played by the Trp/Kyn pathway on the orosensory perception of lipids, a pharmacological inhibition of the first rate-limiting enzyme of the Kyn pathway  was induced in non-operated DIO mice  were used to suture the pyloric vessels along the greater stomach curve. The gastrostomy was performed on the anatomical line from the pyloric region to the cardiac part of the stomach. The incision site was closed with 8.0 Prolene by using a running suture from the gastro-esophageal junction. Eighty percents of the stomach were removed. A special care has been taken to avoid any damage to neural and vascular systems especially at the level of esophagogastric area. The abdominal skin and muscle were closed with 5.0 Prolene interrupted sutures. The controls were sham-operated.The VSG surgical method has been fully described and video-captured previously . BrieflyTo assess the metabolic efficiency of surgery, oral glucose tolerance tests (OGTT) were performed before and after surgery  Figure B. Mice tTwo-bottle choice tests were performed for 12h at the beginning of the dark period on individually housed mice according to previously published procedure . In brie2 area was analyzed to standardize the Fun counting.After sacrificing isoflurane-anaesthetized mice by cervical dislocation, the tongues were removed, washed in 0.09% NaCl and stored in 10% neutral buffered formalin at 4 °C for 2 days. To determine the fungiform papillae density, the tongues were stained for 20 min with a methylene blue solution  and then washed in PBS before observation. A picture of the anterior dorsal tongue was captured using a light macroscope equipped with a camera , and then a 20mmv/v), before storage at −80 °C until the LC-MS analysis. The metabolomic profile of the samples was determined using a high-performance liquid chromatograph adapted to polar compounds  coupled to a high-resolution mass spectrometer . The mass spectra were collected using a high resolving power  by alternating positive and negative ionization modes. The identification of metabolites in humans and mice was performed using their respective mass and retention time according to the Cribiom and Profilomic databases, respectively.Metabolomics analyses were performed by Profilomic SA  on plasma from blood collected at the end of the first experiment  who were candidates for a VSG were selected according to the following exclusion criteria: treatment able to modify taste perception, surgical care of obesity already performed, buccal withering, diabetes (use of hypoglycemic drugs or fasted plasma glucose level >7 mmol/L), smoking habits, chronic inflammatory pathologies.The exploratory human study was realized in patients from the HumanFATaste2 cohort which is described elsewhere . This stAll subjects received detailed information about the study and provided a written consent. To avoid gender-mediated bias , this stTwo complementary investigations were performed 2 weeks before and 6 months after VSG, in the morning, in overnight fasted patients. Firstly, a targeted plasma metabolite analysis was realized by LC-MS/MS. Plasma C-reactive protein (CRP) levels used as inflammatory marker were assayed using human CRP Elisa kit . Secondly, the oral lipid perception threshold was determined using the 3-ascending force-choice (3-AFC) test according to a previously published procedure . In brieFor the metabolomic analysis, a partial-least-squares (PLS) analysis and multiblock hierarchical PLS were performed with SIMCA P + 12  as described ,37. ANOVOral glucose tolerance tests (OGTT) were performed 2 days before and 2 weeks after surgery A. VSG siTo generate new insights into how nutritional obesity disturbs the orosensory perception of dietary lipids, an untargeted analysis of plasma metabolites was undertaken. Nine metabolites were identified as discriminant the VSG-DIO mice from Sham-DIO controls A. The arInterestingly, VSG broadly corrected the altered plasma metabolite signature found in Sham-DIO mice despite that mice were maintained on HFD. The lower plasma mHis, AAA and sebA levels observed in VSG-DIO mice D suggestTo delineate the phenotypic difference between VSG-DIO and Sham-DIO mice, we first performed behavioral tests. We found that the preference for oily solution was lower in Sham-DIO than in Sham-L mice. Interestingly, this phenomenon was partially corrected in VSG-DIO animals suggesting a deleterious effect of obesity beyond the HFD A. Long-tTo further explore whether a change in the peripheral detection system might explain the improvement of fat preference found in the VSG mice, an analysis of the Fun density in the anterior dorsal tongue was undertaken. Consistent to previous published data , a reducSensory innervation is essential for the gustatory papillae renewal . QA in eTo explore this hypothesis, the pharmacological inhibition of the Kyn pathway was performed in non-operated DIO mice. In order to reduce the Trp degradation found in DIO mice, IDO-1 activity was inhibited by addition of 1-methyl TRP (1MT) in the drinking water  Figure A. After Consistent with the literature ,52, the n = 32) 2 weeks before and 6-months after VSG. This procedure led to a massive body weight loss  and of the perception of oily solution in the mouse. Nevertheless, whether these impacts are direct or indirect remains to be determined. Another limitation concerns the functional link between the Kyn pathway and the Fun papillae density which is based on associations rather than causal relationships. This question needs to be confirmed by mechanistic studies. Nevertheless, our data provide a possible mechanistic explanation to the relationship between variation in the Fun number and inflammatory status previously reported by Kaufman and collaborators ,48. It iIn conclusion, this study strongly suggests that Trp/Kyn metabolism is one of regulatory factors in the cross-talk between nutritional obesity, bariatric surgery and taste of fat. A better understanding of molecular mechanisms responsible for obesity-mediated changes of the orosensory perception of energy-dense foods might lead to new nutritional and/or pharmacological strategies facilitating compliance with healthy dietary recommendations useful to avoid a weight rebound after bariatric surgery.


Dysregulation of the glutamatergic system underlies the pathophysiology of depression. Both negative and positive modulation of NMDARs exert rapid and sustained antidepressant effects by reversing the dysregulated glutamatergic system. Research in the past decades has identified key signaling pathways activated by these rapid acting antidepressants. Here, we review the converging signaling mechanisms shared by rapid acting antidepressants and discuss the recent progress on distinct actions of NMDAR antagonists and NMDAR positive modulators to trigger rapid antidepressant actions. Major depressive disorder (MDD) is a debilitating psychiatric disorder and one of the leading causes of disability worldwide. Although the monoaminergic hypothesis of depression dominated the field for several decades, research in the past two decades demonstrates that the glutamatergic system plays a central role in the pathophysiology of MDD . ConventHere, we review previous and current research on the mechanism underlying the actions of rapastinel and compare with ketamine, including both unique convergent mechanisms underlying the actions of these rapid antidepressants. We discuss novel NMDAR-PAMs and their therapeutic potential for treatment of depression and neuropsychiatric disorders.Rapastinel is a threonine-proline-proline-threonine (TPPT) tetrapeptide derived from a hypervariable region cloned and sequenced from the monoclonal antibody B6B216 ,9 ongoinA receptor agonist) was used to inhibit glutamatergic neuronal activation; the results demonstrated that muscimol completely blocked the increase in phospho-ERK and BDNF release induced by either ketamine or rapastinel. Together, these findings indicate that two different rapid acting antidepressants with opposing NMDAR effects share common molecular and cellular mechanisms.Recently, our studies focused on the converging cellular and molecular mechanisms in the antidepressant actions of rapastinel, an NMDAR-PAM as well as the NMDAR antagonist, ketamine. Our previous studies reported that activation of the mTORC1 signaling pathway , through ERK/AKT activation acts as a key underlying convergent mechanism for the fast-acting antidepressant properties of ketamine . In addiin vivo to reveal mechanistic differences between ketamine and rapastinel. This study demonstrated that like ketamine, a single dose of rapastinel rapidly activated mTORC1 pathway in medial prefrontal cortex (mPFC). Infusion of rapamycin, the selective mTORC1 inhibitor, blocked the antidepressant-like behavioral actions of rapastinel, demonstrating a requirement for the mTORC1 pathway. Further, rapastinel increased the number and function of spine synapses in the apical dendritic tuft of layer V pyramidal neurons in the mPFC, an effect also reported after ketamine treatment [in vivo, this study identified similar molecular and cellular mechanisms for ketamine and rapastinel. However, in this study, the authors also reported some differences. Rapastinel is a centrally active, intravenously administered  peptide, so Liu et al. [Liu et al.  further reatment . Therefou et al.  injectedIt is also likely that ketamine and rapastinel could act at NMDARs on different cell types or recruit unknown distinct signaling pathways that converge on ERK or mTOR signaling. Electrophysiological studies also demonstrated that rapastinel significantly increased the frequency of EPSCs to hypocretin, a measure of thalamocortical synapses. However, rapastinel did not increase the frequency of EPSCs to 5-HT, a measure of cortical-cortical response mediated by the 5-HT2A receptor. Previous studies revealed that ketamine increases both hypocretin- and 5-HT induced ESPC frequency ,12,17,18While both rapastinel and ketamine physically target NMDA receptors and share converging molecular signaling mechanisms, the two molecules have opposing actions on NMDARs. Ketamine is a non-competitive NMDAR antagonist while rapastinel acts as an NMDAR-PAM but how both these drugs exert similar rapid and sustained antidepressant effects and activate convergent downstream signaling pathways has been an unanswered question. One of the most accepted hypotheses that addresses this question is that ketamine and rapastinel could modulate NMDARs on different cell types in mPFC. Recent studies from our lab addressed this issue by using a Crerecombinase dependent viral shRNA approach to produce cell type specific knockdown of the GluN2B subunit of NMDARs in different Cre-lines  . GluN2B Ketamine and rapastinel produce rapid and sustained antidepressant effects. Both molecules increase mTORC1, pERK and pAKT, and BDNF release, resulting in increased synaptic number and function that are associated with antidepressant responses . However


Using mid-infrared plasmons to trigger visible surface enhanced Raman spectroscopy signals within a nanocavity represents new opportunities for fundamental investigation of light–matter interaction within quantum regimes, requiring improved sensing capabilities enabled by well-designed nano/microstructures and characterization systems. Using a variety of nanoantenna structures, the incident optical field can be concentrated into deep subwavelength volumes and realize significant localized-field enhancement 2, which is promising to develop enhanced nonlinear optics3, surface photoctalysis5 and surface enhanced spectroscopic sensing technologies7. It is generally believed that smaller gaps between metallic nanopatterns will result in stronger localized field enhancment due to optically driven free electrons coupled within the gap. However, due to the diffraction limit of classic optics, it is a grand challenge to couple and confine the light into these deep subwavelength volumes. Therefore, the capability to manipulate light within these extremely deep subwavelength scales is promising to introduce revolutionary advances in light–matter interaction and pave the way towards light manipulation within extremely small and even quantum regimes8.The unprecedented ability of light concentration within metallic nanostructures has attracted significant research interests in recent years9. Absorption peaks in infrared absorption spectroscopy generally correspond to Raman peaks in SERS spectra. These two technologies are usually considered as complementary technologies that can produce more sensitive capabilities when performed simultaneously. However, since the vibrational absorption signal of surface enhanced infrared absorption spectroscopy is proportional to |E/E0|2 in contrast to |E/E0|4 in SERS, light trapping and localization structures for both infrared wavelengths and visible to near infrared wavelengths are highly desired12. In addition to the development of better light localization nano/micro-structures, another key insight is that it is essential to combine engineered nano/micro-structures with optimized macro-optical systems to boost the overall sensing capability. In particular, a well-designed system may introduce new capabilities to further manipulate the optical field within the tiny volume, which was not possible before. For instance, an earlier example employed two atomic-force microscope tips with metal particles to manipulate the tip-tip distance accurately13. By characterizing the optical scattering spectrum and current simultaneously, a quantum scale limit of 0.3 nm was revealed at which tunneling plasmonics start to dominate, representing the upper limit for localized plasmonic enhancement. With smaller gaps, this enhancement will decrease due to the emergence of quantum-tunneling charge-transfer plasmons, uncovering new areas for quantum plasmonics14 and extreme nonlinear interactions3 within nanometer scales.In a recent issue published in 2021, Wang and Tian et al. overviewed the latest research progress from nano/micro-photonic structures to macro-optical deisngs, and explained their implimentations in surface enhanced Raman spectroscopy (SERS) and infrared absorption spectrocopy technologies15. The authors employed a carefully designed optical system to focus the visible and mid-IR beams to excite plasmons within a designed nanogap and correlated the light–matter interaction in different spectral regimes . As illustrated in Fig. 14). By modulating the mid-IR perturbation signals, the SERS signal at the visible wavelength emitted from the molecules within the gap between the Au sphere and the planar film can be controlled, which is similar to the general principle of optical modulators and/or optical switches. Relying on advanced characterization capabilities , this work observed the time-dependent rise and decay of the SERS signal in a few hundred nano-seconds, opening a way to explore the optical manipulation of SERS signals of trace chemical molecules using mid-IR perturbation.Recentlly, Chikkaraddy and Baumberg reported another example to demonstrate this type of combination: i.e. using mid-infrared (mid-IR) plasmons to trigger visible SERS signals within a nanocavityAlthough the repeatability of this sophisticated characterization needs further improvement, this combined sensing system with nanophotonic structures and advanced detection schemes indicates a new regime for fundamental investigation. Remarkably, it opens the door to manipulate the optical signature of chemical molecules from tiny volumes and study new mechanisms within quantum regimes. It will also enable the exploration of building blocks for efficient light trapping and localized field concentration that will yield important technological breakthroughs for on-chip integrated photonics and optical modulators and sensors in bio/chemical systems.


Lactobacillus species have been studied for over 30 years in their role as commensal organisms in the human gut. Recently there has been a surge of interest in their abilities to natively and recombinantly stimulate immune activities, and studies have identified strains and novel molecules that convey particular advantages for applications as both immune adjuvants and immunomodulators. In this review, we discuss the recent advances in Lactobacillus-related activity at the gut/microbiota interface, the efforts to probe the boundaries of the direct and indirect therapeutic potential of these bacteria, and highlight the continued interest in harnessing the native capacity for the production of biogenic compounds shown to influence nervous system activity. Taken together, these aspects underscore Lactobacillus species as versatile therapeutic delivery vehicles capable of effector production at the lumenal-mucosal interface, and further establish a foundation of efficacy upon which future engineered strains can expand. Lactobacillus species have traditionally maintained positive association with gastrointestinal health. This review summarizes recent research relating to human health associated Lactobacillus administration on cellular and systematic levels. As techniques in molecular and synthetic biology continue to advance, the potential to increase the therapeutic scope and efficacy of Lactobacillus cultures increases regarding target molecule production, immune system influence, vaccination and nervous system activities.Bacteria such as  The rising incidence of systemic diseases is one of the many driving factors for therapeutic development ,2. The ein vitro and in vivo in an effort to elucidate the host-beneficial mechanisms [Lactobacillus, a collection of Gram-positive bacteria many of which are generally regarded as safe food-grade organisms commonly found as commensals in the animal gut microbiota. These bacteria have been studied for both their industrial applications in the production of flavor compounds and other fermentative abilities [Lactobacillus belongs to the larger lactic acid bacteria (LAB) group, for which genetic tools have been developed in efforts to increase their potential as immune adjuvants and cell factories charged with the secretion of bioactive molecules [Lactobacillus-related activity at the gut-microbiota interface such as the immunomodulating effects of these bacteria on immune and epithelial cells. We further highlight the discovery efforts for native compounds produced by these bacteria that can be delivered directly to the epithelia for immunomodulation, as well as the use of recombinant techniques to produce living vaccines to stimulate adaptive immunity. Finally, we discuss recent insights into the role of these bacteria in the production of and influence on native biogenic compounds capable of stimulating the nervous system in order to emphasize the direct and indirect therapeutic potential of these bacteria for the treatment of a diverse set of diseases.The composition of the microbiome is variable and highly elusive, but certain bacterial groups have arisen through study in past decades to be recognized for their probiotic characteristics, wherein health benefits observed in human populations can be correlated to the inclusion of these bacteria in their customary diets . As a rechanisms . Many ofbilities , in addibilities . Lactobaolecules . In effeLactobacillus culture administration can vary depending both on the type utilized and the ailment being studied. The majority of studies follow a formula where a disease model is generated by a well-characterized agonist and then treated with the Lactobacillus culture or derivative. The resulting analysis of cytokine panels and cell surface markers then implicate how the host system responded to the treatment. Different results are desirable depending on the targeted therapeutic avenue. For example, studies on immune system priming aimed at adding protection against pathogen infection would be considered successful with specific T-cell differentiation patterns indicating immune system activation via well-studied Toll-like receptor (TLR) signaling pathways [Lactobacillus species are being studied to treat overactive immunity such as with models of colitis characterized by unchecked inflammation events, the specific changes that signify success are of those that attenuate the host immune response such as increases of the anti-inflammatory cytokines TGF-β, IL-10 and IL-22. The establishment of a disease profile that can be reversed or reduced by the addition of the Lactobacillus component is therefore promising evidence for use of a specific Lactobacillus strain as a living therapeutic.The probiotic properties of LAB have been studied for decades . The stupathways . In thisLactobacillus cultures on cytokine stimulation in a variety of formats. The introduction of Lactobacillus into model systems does not go unnoticed by host immune systems regardless of administration strategy. Considering that cytokines coordinate host responses to different immune events, few exogenous elements can avoid initiating cytokine activity. The innate immune system acts as one of the first lines of defense against pathogen invasion, and its stimulation involves the activation of TLRs [Lactobacillus species, T-cell differentiation will be crudely lumped into the categories of an active form  responsible for the secretion of pro-inflammatory cytokines , and a modulating form (regulatory T or TReg) responsible for regulation of active T cells via secretion of anti-inflammatory cytokines (TGF-β and IL-10). The role of monocytes in directing immune system activity is also an important factor affected by Lactobacillus cultures, specifically dendritic cells (DCs) and macrophages for their TLR signaling tendencies in addition to the T cell differentiating cytokines they produce.A wealth of data in recent years has been collected on the influence of  of TLRs . This ac of TLRs , therefoLactobacilli species on monocytes and naive T cells has been noted in recent years. Investigators have observed interesting changes in immune cell behaviors in the presence of not only the organisms themselves, but also spent culture media and recombinantly produced cell wall components. For example, macrophages produce decreased amounts of TNF-α, IL-1β and IL-17 in the presence of L. plantarum isolates from cocoa fermentation [H cell populations from peripheral blood mononuclear cell (PBMC) cultures in addition to increased amounts of IL-10 from mononuclear cells, indicating the ability of this Lactobacillus culture to both attenuate inflammatory signaling through direct monocyte contact and influence T-cell maturation in mixed cultures. These results are not uncommon in this field, as many investigators have noted similar changes in monocyte behavior as well as T-cell maturation. Other examples include the increase in TH1 populations in high-fat diet mouse models caused by L. rhamnosus and L. plantarum [reg populations in enteritis mouse models with L. casei [L. paracasei and L. reuteri [L. plantarum [L. rhamnosus with IgA [Lactobacilli on an organismal scale, wherein the skewing of T-cell populations and DC activity to a host-beneficial profile is detailed more thoroughly in the individual studies.The effect of different entation . The samlantarum , the incL. casei  and in s reuteri . Effectslantarum  and L. rwith IgA . StudiesLactobacilli cultures found interesting results regarding their adhesion properties with respect to the phagocytotic activity of macrophages. The pilus encoded by the spaCBA genes from L. rhamnosus strain GG (LGG) was recently found to be integral for direct macrophage contact resulting in anti-inflammatory TLR2 signaling [L. casei cultures [spaCBA deletion mutants of the spaCBA pilus did not have the same effect on signaling [L. acidophilus cultures using DCs, where investigators narrowed the cause down to alternate signaling processes dependent on activation of pathways downstream of TLR2 itself. The data showed that upon uptake of L. acidophilus, endosomal degradation as well as downstream activation of pro-inflammatory genes such as IFN-β was prevented [Lactobacilli inducing both pro-inflammatory [Lactobacillus cultures and monocytes [Investigators attempting to identify more specific molecular mechanisms responsible for the immune cell priming and immunomodulation by ignaling . Studiescultures . These rignaling . Similarrevented . While tammatory ,30 and sammatory , the indonocytes ,29 demonLactobacillus on mammalian cells involves increasing anti-inflammatory activity. While inflammation is an integral part of immunity, there are a number of diseases characterized by unchecked inflammation that damages host tissue [Reg cells, and some cases of inflammation damage occur when these populations and associated signaling pathways are out of balance from unknown external or auto-induced stimuli. The anti-inflammatory influences of Lactobacillus species therefore provide a therapeutic strategy with minimal impact.The most established effect of t tissue . FurtherLactobacillus species on immune cell maturation and differentiation discussed above accounts for one facet of their potential therapeutic scope. In addition to their effects on naive lymphocytes, different Lactobacillus species can stimulate anti-inflammatory cytokine production in a number of different scenarios  or 2,4,5-trinitrobenzenesulfonic acid (TNBS) are characterized by high levels of pro-inflammatory cytokines , and upon the administration of Lactobacillus there are trends of increased IL-10 and TGF-β as well as decreased pro-inflammatory markers similar to that seen with cell culture experiments. One particular study showed the anti-inflammatory effects described above with L. sakei food isolates as comparable with 50 mg/kg of the anticolitic drug sulfasalazine [Lactobacillus cultures reduced TNF-α and IL-1β by 72.6 and 52.9%, respectively, and increased IL-10 to 56.1% in TNBS-induced mouse models. These effects exemplify the potential viability of this anti-inflammatory therapy, as there was no significant difference between Lactobacillus and sulfasalazine treatments [et al. with L. acidophilus, decreases in IL-17, IL-23 and TGF-β were noted upon treatment of the inflammation model. Similar to previously mentioned cell culture studies where observed cytokine changes are potentially confusing, this is a case where both pro- and anti-inflammatory cytokines decrease. However, the investigators were able to utilize the wealth of available data to conclude these changes were caused by decreases in TH17 populations, and they ultimately represent an anti-inflammatory response. While these results are not typical among recent studies, they represent the clarity that animal studies bring to otherwise complex issues resulting from therapeutic assessments. Similarly, many recent studies have used the wealth of data accumulated from animal studies to conclude anti-inflammatory outcomes from Lactobacillus therapies in colitis [K. pneumoniae infection models [Animal studies can be associated with variability and difficulty assigning contributions of specific cell sources, but they are far superior for observing the broad therapeutic outcomes of salazine . In thiseatments . As ment colitis ,43,45, e colitis  and K. pn models .Lactobacillus cultures that offer intriguing results. One particular study involved L. casei administration to healthy adults, from whom macrophages and T cells were then extracted for ex vivo studies [Lactobacillus species for an anti-inflammatory therapy. Adults with metabolic syndrome were administered L. reuteri in a separate study, wherein microbiome composition was altered in addition to decreases in IL-6 and soluble vascular adhesion molecule 1 as metabolic syndrome symptom markers [L. plantarum supplements revealed similar results as in mouse studies with decreases in plasma IFN-γ and TNF-α in addition to increases in IL-10 [Lactobacilli species was shown to enhance immunomodulation and thus prevent subsequent associated tissue damage [There are a few recent human studies involving the effects of  studies . Investi markers . Studiesin IL-10 . The impe damage .Lactobacilli on immune functions is intriguing, especially when considering a therapeutic strategy to combat a systemic inflammatory disorder. Interactions of Lactobacillus cultures with epithelial cells comprise another aspect of this therapeutic potential given their role as a physical barrier between lumenal and humoral spaces. As epithelial layers can act as avenues of pathogen invasion in the respiratory, urogenital and gastrointestinal tracts, the influence of Lactobacillus cultures on barrier maintenance and their protective effects against pathogen adhesion and invasion are two points of interest for this review.The effect of Lactobacillus species, these bacteria similarly are involved in the maintenance of intestinal barrier integrity.The gut microbiome has a complex role in its interactions with host nervous and immune systems, but the benefits of this relationship are tethered to the gut itself being a self-contained environment. The intestinal barrier therefore protects the bloodstream from the diverse gut microbiome residing in the lumenal spaces . While tLactobacillus has existed for decades and is summarized in Lactobacillus species were used to reverse the effects of pathogen- or chemical-induced breakdown of barrier functions. The majority of studies show that Lactobacillus treatments maintain trans-epithelial electrical resistance measurements in the face of barrier antagonists as well as decrease pro-inflammatory cytokine concentrations. The effects of Lactobacillus treatment on TJP expression are also noted in Lactobacillus species in both cell culture and animal studies. Lactobacillus treatment including maintenance of the proteins ZO-1 (a TJP) and MUC2 (involved in mucus layer integrity) via differential staining microscopy  infection. These results act as a reminder of the interplay between multiple cell types as well as the importance and benefit of studying diverse co-culture systems in addition to animal models. As mentioned above, utilizing multiple cell types increases the complexity of the data, but it allows for a broader assessment of therapeutic efficacy.One particularly interesting study involved the use of tion NKs . Primed Lactobacillus species to host cells can influence immune signaling as described above, it also efficiently blocks pathogen adhesion and subsequent tissue damage. The adhesion characteristics of Lactobacillus have recently been studied in a couple of contexts. In one example, the purified adhesin proteins MUB and CmbA from L. reuteri were shown to upregulate IFN-γ secretion from TH1 cells as well as IL-10, TNF-α, IL-6 and IL-12 from monocyte-derived DCs [L. rhamnosus GG with epithelial cells [Lactobacillus pretreatment has also been used to prevent pathogen invasion in a number of studies  and exopolysaccharides (EPS), however, are still under investigation  that are both strain- and environment-specific . While sirulence ,99–103. irulence  and act irulence , with thirulence . Investiirulence ,114–116.irulence , pro-infirulence ,118,119 irulence ,120. Simirulence ,122, theLactobacillus species possess a number of characteristics that make them ideal vehicles for biomolecule delivery. In addition to their abilities to influence the immune system described above, these native microbiome constituents can survive the harsh gastrointestinal tract and have subsequently been the focus of engineering efforts [Lactobacillus species offer tunability in delivery for the adjustment of therapeutic contact time, exposure and clearance from host systems.In fact,  efforts ,123 for  efforts ,125 in a efforts ,127, LacLactococcus lactis secretion of bioactive IL-10, effectively modulating inflammatory symptoms in colitis mouse models [Lactobacillus species to prime adaptive immunity [LAB have been used for mucosal delivery of therapeutics for decades. Initial efforts in this field demonstrated methods for e models , and sube models  and dispe models  of IL-10e models , TGF-β1 e models  and serie models . Interesimmunity  in orderLactobacillus species present an ideal balance between immunogenicity and reactogenicity to efficiently achieve immune activation  such as acetate, butyrate [LAB have been known to generate or influence host concentrations of biogenic compounds for some time. Many of these are decarboxylated amino acids such as tyramine, tryptamine, histamine or γ-aminobutyric acid (GABA) that have various effects on the host nervous system. While histamine has been well studied in the context of immunity for its activity during inflammation when released by mast cells, tryptamine and tyramine (among others) are TAAR1 agonists and GABA is known to act as an integral inhibitory neurotransmitter in the human adult brain. The actions of the affected signaling pathways and their physiological implications have been reviewed recently , castingbutyrate  and propbutyrate  in the gbutyrate  and actibutyrate  dependinLactobacillus species to produce SCFAs, investigators have recently examined the effect of different bacterial species on SCFA homeostasis in animal models. Studies have noted alterations in SCFA abundances and ratios in addition to the associated anti-inflammatory benefits with L. rhamnosus [L. plantarum [L. johnsonii [L. sakei [, L. reuteri [L. acidophilus [L. paracasei [L. plantarum [Lactobacillus cultures specifically producing SCFAs and influencing host cell surface receptor expression, the actual homeostasis is a complicated dance within the gut that both reflects and sculpts the community structure. Some of the nuances to this system include the effects of antimicrobial peptides (bacteriocins) produced by LAB, capable of killing specific groups of bacteria thereby indirectly altering SCFA ratios [Based on previous observations of the abilities for different hamnosus , L. planlantarum ,169, L. ohnsonii , L. sakeL. sakei , L. reut reuteri  and L. adophilus . Similararacasei  and L. plantarum . Deeper lantarum ,177 despA ratios ,179. DesAdministering therapeutics that influence neurotransmitter activity is an intriguing subject, but there are always associated concerns of efficacy, dosage and safety. Considering the complexity of the human nervous system, one question as to the feasibility of bacterial production is whether the generated doses can produce the desired outcome. In the case of GABA, the gut represents a large pool in its generation, consumption, and activity , therefoLactobacillus brevis has been considered a high-capacity GABA-producing organism for over a decade, shown to convert up to 30 mM of L-monosodium glutamate (MSG) to GABA (~3 g/l) in 4 h [L. brevis has led to its popularity in animal testing to discern whether the production in situ can effect measurable change in neural activity. Recent animal studies have shown that this culture can influence activity in the vagus nerve through serotonin 5-HT3 receptors [Lactobacillus administration to different diseases associated with nervous system and hormone signaling. One such disease targeted for study is Type 2 Diabetes, considering the role GABA plays in glucagon release from β cells, and as such investigators have shown L. brevis to positively affect glucose reduction in diabetic animal models [In vitro studies with activated immune cells have also demonstrated immunomodulatory capabilities of these GABA-producing cultures [) in 4 h . While t) in 4 h , this checeptors , openingl models ,186. In cultures .Lactobacillus cultures have also been shown to affect neural activity independent of GABA production. These cases are less determinative without a specific molecule to assign responsibility, but the results are nevertheless intriguing. In studies with both cell culture [Lactobacillus species were shown to cause increases in brain-derived neurotrophic factor that were positively correlated with cognitive activity [ culture ,189 and  culture ,191, Lacactivity ,191. TheLactobacillus have provided increased evidence of their potential as living therapeutic delivery systems. Various species have been shown to influence T-cell differentiation patterns reflecting both immune-priming and immunomodulating pathways in different scenarios. Specific interactions with monocytes have identified signaling pathways initiated by different bacteria to uncover a differential phagocytic end point among bacterial sources, with the immune cells appearing to differentiate between bacterial species. These revelations comprise the foundation of results echoed throughout a wealth of cell culture and animal model studies that overwhelmingly support the role of the Lactobacillus species as enhancers of immune regulatory activity. When administered as a pretreatment, investigators were able to take advantage of immune-priming characteristics of some strains to stimulate the immune system and initiate antibody generation, thereby enhancing pathogen protection in a long-term prophylactic way as a living vaccine delivery system. While many of the effects of Lactobacillus are caused by factors yet to be elucidated, studies have shown specific culture products such as LTA, EPS, surface layer proteins, SCFAs and GABA to display a variety of effects on both cellular and organismal levels. The potential to deliver a variety of therapeutic options is high with Lactobacillus, especially when considering cooperative activity from multiple species. An example of this scope is shown with the improvement of disease symptoms in diabetic models caused by Lactobacillus production of GABA in addition to similar strains being used to reduce tissue damage from pathogen-induced inflammation and improve cognitive responses [The previous 5 years of research with esponses .Lactobacillus [Lactobacillus species to further optimize therapeutic strategies. These longer term goals will be difficult and could require extensive metabolic engineering to produce, given the complexity of strain-specific LTA and EPS modifications.The optimal understanding of the underlying mechanisms has yet to be reached in order to support the universal deployment of such therapeutics. Many of the tested species have been highly studied with very promising results, but it is hard to imagine these therapeutic strategies as being welcomed by the medical field without further investigations into the identity and molecular mechanisms of species-specific effector molecules to understand and further optimize their therapeutic potential . While sbacillus . The speL. rhamnosus GG adhesion protein SpaC to enhance interactions with host cells. While the potential for widespread therapeutic use is great, any incremental step toward these goals requires confirmation from parallel cell culture and animal studies to assess the organismal impact of any treatments. A number of studies reviewed here have done such parallel studies (Lactobacillus administration for the production and localized delivery of native bioactive compounds.While a multitude of naturally isolated species have been shown to be effective, there are a number of opportunities available to improve therapeutic efficacy given the identification of necessary mechanisms. As the composition of the gut microbiome has been correlated with therapeutic outcomes, novel antimicrobial peptides generated by recently pioneered machine learning methods  could be studies , but accLactobacillus species as widely accepted and commonly used therapeutic delivery vehicles is an attainable goal. The tools required for genetic manipulation, cultivation, isolation and testing are already available in most molecular biology labs today. The avenue of vaccine delivery is already a streamlined laboratory workflow whose primary current limitation is the selection of antigens to recombinantly express for the Lactobacillus-facilitated exposure to subjects. Without the next generation engineering goals of altering metabolism, adding adhesion properties and generating novel gut community structuring strategies mentioned above, there are still a number of well-studied and readily available strains that have been demonstrated capable of treating various immune and neural-centered abnormalities. The establishment of a next generation of Lactobacillus strains would solidify their reputation as therapeutic delivery vehicles with accessible capabilities for scaled-up production and modification. These attributes would allow treatment strategies to quickly evolve as new research emerges regarding the gut microbiome influence on host neural and immune behaviors.The successful implementation of Lactobacillus strains have been demonstrated in numerous studies to regulate inflammation, protect the epithelia from pathogen invasion, reinforce the intestinal barrier through the influence of tight junction proteins and stimulate the adaptive immune system.Microbial engineering strategies have emerged to modify these bacteria for the production of bioactive compounds to supplement native probiotic activities.Lactobacillus species to display antigens and act as living vaccines that efficiently stimulate specific adaptive immunity through interaction with the gut mucosa.Further protein engineering strategies have been used to decorate Lactobacillus-influenced levels of neurotransmitters and short-chain fatty acids highlight the potential for disease modulation as shown in animal models.Links between different disease progression mechanisms and the Lactobacillus species are emerging synthetic biology platforms that can be paired with machine learning combinatorial methods for bacteriocin production to beneficially sculpt gut microbiome composition for SCFA homeostasis, pathogen control and reinforced immunity.


ABSTRACT IMPACT: This proof-of-concept study demonstrates that systematically altering limb dynamics with two exoskeleton devices alters ingrained, bilateral upper-limb coordination with the potential to rehabilitate functional reaching in chronic stroke survivors OBJECTIVES/GOALS: Advances in virtual reality and exoskeleton technologies have allowed researchers to alter upper limb coordination with more precision than ever before. The goal of this study was to systematically enhance the use of the nondominant limb during a bimanual reaching task, with an eye towards improving rehabilitative strategies post stroke. METHODS/STUDY POPULATION: Healthy, right-handed volunteers performed a bimanual reaching task in virtual reality (VR) space while simultaneously moving under the influence of two exoskeleton devices. The VR task had participants move a shared cursor, displayed at the midpoint between the hands, to targets arranged at shoulder and eye levels and located at 70% of full arm extension. Two exoskeleton devices applied either resistive torque to the dominant limb or assistive torque to the non-dominant limb. Three-dimensional hand position data were recorded at 50 Hz and analyzed offline. The primary outcome measure was relative contribution, calculated as the ratio of dominant/non-dominant displacement. RESULTS/ANTICIPATED RESULTS: Preliminary results from 3 participants showed that during baseline trials, when no torque was applied by the exoskeletons, relative contribution was 50.6% in favor of the dominant hand, with the dominant hand reaching on average 1.1cm farther than the left. When the exoskeletons resisted movement in the dominant limb while simultaneously assisting movement in the non-dominant limb, relative contribution was 49.7% indicating an increase in non-dominant limb usage. Further analysis showed that this effect was driven by one participant who reached 3.7cm farther with her non-dominant hand compared to baseline. DISCUSSION/SIGNIFICANCE OF FINDINGS: These pilot data suggest our testing platform is capable of altering normal coordination patterns and is likely the result of participants adopting an optimal control strategy imposed by the shared cursor. These findings will form the basis for a rehabilitation intervention to promote the use of the paretic limb in chronic stroke survivors.


The Omicron variant of SARS-CoV-2 is emerging in communities where people were previously infected with SARS-CoV-2 and are now being vaccinated, or where many people have received two or three coronavirus vaccination doses. More than 130 countries around the globe have implemented booster dose programs for tackling omicron endemics. Despite early findings shows that booster doses may improve omicron protection, more research is needed to establish vaccination efficacy. This short communication tries to critically discuss the research work findings around booster dose strategy for omicron endemics. Booster dose is to boost the immune response sometime later after primary immunization. In the case of coronavirus disease 2019 (COVID-19), it was recommended to have 2 injections of most of the vaccines, and 6 months later they are seeing a decline in immunity . As per The WHO named the newly categorized variant of concern (VOC), the Omicron variant, a “severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)”, due to its ominous earlier prestige . First dThe supplemental dose of mRNA vaccines now distributed may result in more rigorous and long-lasting host defense, precluding the need for additional boosters. Furthermore, the development of novel vaccines that provide protection against less mutable coronavirus proteins may result in broader protection . AdditioMore than 130 countries around the globe have implemented booster dose programs, although the coverage rates for complete primary vaccination are below 30% in the majority of these countries. The major goal to halt this pandemic is to achieve 100% primary vaccination coverage globally with a secondary objective of full vaccination of elderly and immunocompromised individuals for a substantial reduction in severe disease and mortality. Given the ongoing logistic challenges in global vaccine availability and equity, each nation’s vaccine-booster dosage-policy decisions must balance public health advantages for their populations with support for global equality in vaccine access, which is required to address virus mutation and the effect of the pandemic. Indications of decreasing vaccine efficacy, namely a loss in coverage against severe illness in high-risk groups, necessitates the development of vaccination techniques tailored for severe disease prevention, including the tailored use of booster immunization.


We present a 62-year-old gentleman with rapidly forming abdominal pseudoaneurysms due to segmental arterial mediolysis (SAM). With rupture of his pseudoaneurysms, he underwent angiography and successful coil embolisation. In this case, we demonstrate the potential for rapid progression of pseudoaneurysms in SAM, with the need for prompt diagnosis and urgent endovascular intervention. SAM most commonly affects coeliac, mesenteric and renal arteries, and its presentation varies from chronic mesenteric ischaemia to acute life-threatening haemorrhage thus posing both a diagnostic and therapeutic challenge. We report a case of SAM in a 62-year-old gentleman presenting with rapid formation and rupture of pseudoaneurysms who was successfully treated with early coil embolisation.Segmental arterial mediolysis (SAM) is an uncommon but critical condition with up to a 60% mortality rate in the acute phase from catastrophic hemorrhage. It is a non-inflammatory, non-atherosclerotic vasculopathy with segmental lysis of the tunica medial layer of abdominal arteries.A 62-year old gentleman presented with sudden onset, severe generalised abdominal pain. He had normal vitals signs but on examination demonstrated generalised peritonism. Biochemistry showed lactataemia of 4.2 mmol/L but was otherwise unremarkable. Computer tomography (CT) imaging of his abdomen was equivocal for an early superior mesenteric artery dissection. Spontaneous and complete resolution of the patient’s symptoms occurred within an hour of presentation and he was admitted for monitoring. Twenty-four hours later the patient experienced the same severe, sudden onset, abdominal pain accompanied by hypotension and syncope. A CT mesenteric angiogram demonstrated two pseudoaneurysms arising from the splenic artery with associated rupture of the proximal pseudoaneurysm and retroperitoneal haemorrhage  and a low suspicion of alternative arteriopathies . Negative inflammatory markers such as antinuclear antibodies and antineutrophil cytoplasmic antibodies contributed to the diagnosis of SAM. However, tissue diagnosis, if obtainable, can assist in differentiating mimics of SAM include anti-neutrophil cytoplasmic autoantibody-associated vasculitis, polyarteritis nodosa, C and mycotic aneurysmal infections. In cases where the early phase of SAM when the diagnosis is unclear, a combination of tissue diagnosis, vasculitis biomarkers and infection screens can be useful in distinguishing similar arteriopathies.SAM is a rare vascular disorder with unknown aetiology first described by Slavin and Gonzalez-Vitale in 1976. Shenouda et al. reviewed 85 cases of SAM with 24 cases utilising endovascular intervention and 36 cases undergoing emergency open laparotomy. Open surgery was associated with poorer outcomes with 9% mortality compared to 0% in those undergoing endovascular treatment. However, open surgery is valuable in the event of failed endovascular intervention or in the elective setting for vessel ligation for known stable pseudoaneurysms. Alternatively, patients who survived the acute phase of the disease can be managed medically with anti-hypertensives, antiplatelets or anticoagulation in combination with serial CT imaging. Fortunately, disease regression and stability were observed in many patients after resolution of their initial acute episode. Several studies have concluded after the acute phase, majority of patients demonstrated regression or stability of disease.Treatment for SAM in the acute setting includes emergent endovascular intervention, open surgery or medical management.Vascular surgeons should be aware of SAM as a differential for patients presenting with acute abdominal pain. SAM as a life-threatening condition requiring the need for early angiography and embolization via interventional radiology, rather than diagnostic open exploratory laparotomy. This case contributes to our understanding of the variability in SAM’s clinical presentation and knowledge of its potential to rapidly propagate pseudoaneurysms at risk of rupture. It emphasises the need for vigilance when approaching abdominal pain with initially unclear aetiology.


Pseudomonas phages and provides a selective advantage when these replicate in some host strains. Throughout the lytic cycle, the phage nucleus compartmentalizes proteins according to function and protects the phage genome from host defense mechanisms. Early during infection, the PhuZ spindle positions the newly formed phage nucleus at midcell and, later in the infection cycle, the spindle rotates the nucleus while delivering capsids and distributing them uniformly on the nuclear surface, where they dock for DNA packaging. During the co-infection of two different nucleus-forming jumbo phages in a bacterial cell, the phage nucleus establishes Subcellular Genetic Isolation that limits the potential for viral genetic exchange by physically separating co-infection genomes, and the PhuZ spindle causes Virogenesis Incompatibility, whereby interacting components from two diverging phages negatively affect phage reproduction. Thus, the phage nucleus and PhuZ spindle are defining cell biological structures that serve roles in both the life cycle of nucleus-forming jumbo phages and phage speciation.Bacteriophages and their bacterial hosts are ancient organisms that have been co-evolving for billions of years. Some jumbo phages, those with a genome size larger than 200 kilobases, have recently been discovered to establish complex subcellular organization during replication. Here, we review our current understanding of jumbo phages that form a nucleus-like structure, or “Phage Nucleus,” during replication. The phage nucleus is made of a proteinaceous shell that surrounds replicating phage DNA and imparts a unique subcellular organization that is temporally and spatially controlled within bacterial host cells by a phage-encoded tubulin (PhuZ)-based spindle. This subcellular architecture serves as a replication factory for jumbo  Mimiviridae contain actin-related genes, called viractins  , 2019. AWhen procapsids reach the phage nucleus at the depolymerizing plus end of the filament, they dock onto the nuclear surface and initiate the DNA packaging process. As the incoming procapsid arrives at the same subcellular location as the previous procapsid, the PhuZ spindle also rotates the phage nucleus when the two filament ends of the spindle from each side push the structure transversely in order to provide a new surface for docking . This roPseudomonas PhiKZ-like viruses: 201Phi2-1, PhiPA3, and PhiKZ . The phage nucleus thus establishes “Subcellular Genetic Isolation,” a condition limiting the opportunity for co-infecting viruses to recombine due to subcellular spatial restrictions (Viruses are known to have very high rates of genetic exchange that can occur when two phages infect the same host cell . Recent rictions . Subcellrictions . For exarictions , poxvirurictions  and nuclrictions , assemblrictions . Thus, fAnother mechanism that limits the potential for genetic exchange between jumbo phages is “Virogenesis Incompatibility,” in which components from two distinct phages interact to negatively affect phage reproduction . We haveA second Virogenesis Incompatibility factor is the PhiPA3 protein gp210, a nuclease which normally resides within the PhiPA3 nucleus. Transporting PhiPA3 gp210 into the PhiKZ nucleus greatly reduces PhiKZ production, indicating the incompatibility of this protein with the production of PhiKZ particles. PhiPA3 gp210 is a Virogenesis Incompatibility factor that could contribute to the divergence of PhiPA3 and PhiKZ by specifically reducing PhiKZ production under conditions in which PhiPA3 and PhiKZ formed a single, shared nucleus during co-infection . Thus, tSimilar to Subcellular Genetic Isolation, we expect Virogenesis Incompatibility to also be widespread among both eukaryotic and prokaryotic viruses. Since presumably all viruses require the production of virion particles from a limited set of self-assembling components, such as capsids, tails, tail fibers, portals, terminases, nuclear shells, PhuZ spindles, etc., divergence between any of these polymerizing proteins can potentially result in Virogenesis Incompatibility during co-infection, as we have demonstrated for the PhuZ spindle. Virogenesis Incompatibility has also been reported for eukaryotic segmented viruses: influenza A and B . ConsistThe eukaryotic nucleus is the defining structure of eukaryotic cells. It is a membrane-bound organelle with a lamina protein scaffold inside that separates genomic DNA from the cytoplasm and serves the key purpose of uncoupling transcription and translation . RecentlVC, EB, and JP contributed on the conception and flow of this minireview, wrote the draft, revised the manuscript, and approved it for publication. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.