Shuffling of Promoters for Multiple Genes To Optimize Xylose Fermentation in an Engineered Saccharomyces cerevisiae Strain▿ †

We describe here a useful metabolic engineering tool, multiple-gene-promoter shuffling (MGPS), to optimize expression levels for multiple genes. This method approaches an optimized gene overexpression level by fusing promoters of various strengths to genes of interest for a particular pathway. Selection of these promoters is based on the expression levels of the native genes under the same physiological conditions intended for the application. MGPS was implemented in a yeast xylose fermentation mixture by shuffling the promoters for GND2 and HXK2 with the genes for transaldolase (TAL1), transketolase (TKL1), and pyruvate kinase (PYK1) in the Saccharomyces cerevisiae strain FPL-YSX3. This host strain has integrated xylose-metabolizing genes, including xylose reductase, xylitol dehydrogenase, and xylulose kinase. The optimal expression levels for TAL1, TKL1, and PYK1 were identified by analysis of volumetric ethanol production by transformed cells. We found the optimal combination for ethanol production to be GND2-TAL1-HXK2-TKL1-HXK2-PYK1. The MGPS method could easily be adapted for other eukaryotic and prokaryotic organisms to optimize expression of genes for industrial fermentation

Biomechanical and Clinical Study of Rod Curvature in Single-Segment Posterior Lumbar Interbody Fusion

Objective: Pedicle screw fixation is a common technique used in posterior lumbar interbody fusion (PLIF) surgery for lumbar disorders. During operation, rod contouring is often subjective and not satisfactory, but only few studies focused on the rod-contouring issue previously. The aim of the study was to explore the effect of the rod contouring on the single-segment PLIF by the finite element (FE) method and retrospective study. Methods: A FE model of the lumbosacral vertebrae was first reconstructed, and subsequently single-segmental (L4/5) PLIF surgeries with four rod curvatures (RCs) were simulated. Herein, three RCs were designed by referring to centroid, Cobb, and posterior tangent methods applied in the lumbar lordosis measurement, and zero RC indicating straight rods was included as well. Clinical data of patients subjected to L4/5 segmental PLIF were also analyzed to verify the correlation between RCs and clinical outcome. Results: No difference was observed among the four RC models in the range of motion (ROM), intersegmental rotation angle (IRA), and intradiscal pressure (IDP) under four actions. The posterior tangent model had less maximum stress in fixation (MSF) in flexion, extension, and axial rotation than the other RC models. Patients with favorable prognosis had larger RC and positive RC minus posterior tangent angle (RC-PTA) of fused segments with respect to those who had poor prognosis and received revision surgery. Conclusion: All RC models had similar biomechanical behaviors under four actions. The posterior tangent-based RC model was superior in fixation stress distribution compared to centroid, Cobb, and straight models. The retrospective study demonstrated that moderate RC and positive RC-PTA were associated with better postoperative results.</p

Open-Vocabulary 3D Detection via Image-level Class and Debiased Cross-modal Contrastive Learning

Current point-cloud detection methods have difficulty detecting the open-vocabulary objects in the real world, due to their limited generalization capability. Moreover, it is extremely laborious and expensive to collect and fully annotate a point-cloud detection dataset with numerous classes of objects, leading to the limited classes of existing point-cloud datasets and hindering the model to learn general representations to achieve open-vocabulary point-cloud detection. As far as we know, we are the first to study the problem of open-vocabulary 3D point-cloud detection. Instead of seeking a point-cloud dataset with full labels, we resort to ImageNet1K to broaden the vocabulary of the point-cloud detector. We propose OV-3DETIC, an Open-Vocabulary 3D DETector using Image-level Class supervision. Specifically, we take advantage of two modalities, the image modality for recognition and the point-cloud modality for localization, to generate pseudo labels for unseen classes. Then we propose a novel debiased cross-modal contrastive learning method to transfer the knowledge from image modality to point-cloud modality during training. Without hurting the latency during inference, OV-3DETIC makes the point-cloud detector capable of achieving open-vocabulary detection. Extensive experiments demonstrate that the proposed OV-3DETIC achieves at least 10.77 % mAP improvement (absolute value) and 9.56 % mAP improvement (absolute value) by a wide range of baselines on the SUN-RGBD dataset and ScanNet dataset, respectively. Besides, we conduct sufficient experiments to shed light on why the proposed OV-3DETIC works

Fermentation kinetics for xylitol production by a Pichia stipitis D-Xylulokinase mutant previously grown in spent sulfite liquor

Spent sulfite pulping liquor (SSL) contains lignin, which is present as lignosulfonate, and hemicelluloses that are present as hydrolyzed carbohydrates. To reduce the biological oxygen demand of SSL associated with dissolved sugars, we studied the capacity of Pichia stipitis FPL-YS30 (xyl3 Delta) to convert these sugars into useful products. FPL-YS30 produces a negligible amount of ethanol while converting xylose into xylitol. This work describes the xylose fermentation kinetics of yeast strain P.stipitis FPL-YS30. Yeast was grown in rich medium supplemented with different carbon sources: glucose, xylose, or ammonia-base SSL. The SSL and glucose-acclimatized cells showed similar maximum specific growth rates (0.146 h(-1)). The highest xylose consumption at the beginning of the fermentation process occurred using cells precultivated in xylose, which showed relatively high specific activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49). However, the maximum specific rates of xylose consumption (0.19 g(xylose)/g(cel) h) and xylitol production (0.059 g(xylitol)/g(cel) h) were obtained with cells acclimatized in glucose, in which the ratio between xylose reductase (EC 1.1.1.21) and xylitol dehydrogenase (EC 1.1.1.9) was kept at higher level (0.82). In this case, xylitol production (31.6 g/l) was 19 and 8% higher than in SSL and xylose-acclimatized cells, respectively. Maximum glycerol (6.26 g/l) and arabitol (0.206 g/l) production were obtained using SSL and xylose-acclimatized cells, respectively. The medium composition used for the yeast precultivation directly reflected their xylose fermentation performance. The SSL could be used as a carbon source for cell production. However, the inoculum condition to obtain a high cell concentration in SSL needs to be optimized

Biomechanical and Clinical Study of Rod Curvature in Single-Segment Posterior Lumbar Interbody Fusion

Objective: Pedicle screw fixation is a common technique used in posterior lumbar interbody fusion (PLIF) surgery for lumbar disorders. During operation, rod contouring is often subjective and not satisfactory, but only few studies focused on the rod-contouring issue previously. The aim of the study was to explore the effect of the rod contouring on the single-segment PLIF by the finite element (FE) method and retrospective study. Methods: A FE model of the lumbosacral vertebrae was first reconstructed, and subsequently single-segmental (L4/5) PLIF surgeries with four rod curvatures (RCs) were simulated. Herein, three RCs were designed by referring to centroid, Cobb, and posterior tangent methods applied in the lumbar lordosis measurement, and zero RC indicating straight rods was included as well. Clinical data of patients subjected to L4/5 segmental PLIF were also analyzed to verify the correlation between RCs and clinical outcome. Results: No difference was observed among the four RC models in the range of motion (ROM), intersegmental rotation angle (IRA), and intradiscal pressure (IDP) under four actions. The posterior tangent model had less maximum stress in fixation (MSF) in flexion, extension, and axial rotation than the other RC models. Patients with favorable prognosis had larger RC and positive RC minus posterior tangent angle (RC-PTA) of fused segments with respect to those who had poor prognosis and received revision surgery. Conclusion: All RC models had similar biomechanical behaviors under four actions. The posterior tangent-based RC model was superior in fixation stress distribution compared to centroid, Cobb, and straight models. The retrospective study demonstrated that moderate RC and positive RC-PTA were associated with better postoperative results.</p

Biomechanical and Clinical Study of Rod Curvature in Single-Segment Posterior Lumbar Interbody Fusion

Objective: Pedicle screw fixation is a common technique used in posterior lumbar interbody fusion (PLIF) surgery for lumbar disorders. During operation, rod contouring is often subjective and not satisfactory, but only few studies focused on the rod-contouring issue previously. The aim of the study was to explore the effect of the rod contouring on the single-segment PLIF by the finite element (FE) method and retrospective study. Methods: A FE model of the lumbosacral vertebrae was first reconstructed, and subsequently single-segmental (L4/5) PLIF surgeries with four rod curvatures (RCs) were simulated. Herein, three RCs were designed by referring to centroid, Cobb, and posterior tangent methods applied in the lumbar lordosis measurement, and zero RC indicating straight rods was included as well. Clinical data of patients subjected to L4/5 segmental PLIF were also analyzed to verify the correlation between RCs and clinical outcome. Results: No difference was observed among the four RC models in the range of motion (ROM), intersegmental rotation angle (IRA), and intradiscal pressure (IDP) under four actions. The posterior tangent model had less maximum stress in fixation (MSF) in flexion, extension, and axial rotation than the other RC models. Patients with favorable prognosis had larger RC and positive RC minus posterior tangent angle (RC-PTA) of fused segments with respect to those who had poor prognosis and received revision surgery. Conclusion: All RC models had similar biomechanical behaviors under four actions. The posterior tangent-based RC model was superior in fixation stress distribution compared to centroid, Cobb, and straight models. The retrospective study demonstrated that moderate RC and positive RC-PTA were associated with better postoperative results.</p

Open-Vocabulary Point-Cloud Object Detection without 3D Annotation

The goal of open-vocabulary detection is to identify novel objects based on arbitrary textual descriptions. In this paper, we address open-vocabulary 3D point-cloud detection by a dividing-and-conquering strategy, which involves: 1) developing a point-cloud detector that can learn a general representation for localizing various objects, and 2) connecting textual and point-cloud representations to enable the detector to classify novel object categories based on text prompting. Specifically, we resort to rich image pre-trained models, by which the point-cloud detector learns localizing objects under the supervision of predicted 2D bounding boxes from 2D pre-trained detectors. Moreover, we propose a novel de-biased triplet cross-modal contrastive learning to connect the modalities of image, point-cloud and text, thereby enabling the point-cloud detector to benefit from vision-language pre-trained models,i.e.,CLIP. The novel use of image and vision-language pre-trained models for point-cloud detectors allows for open-vocabulary 3D object detection without the need for 3D annotations. Experiments demonstrate that the proposed method improves at least 3.03 points and 7.47 points over a wide range of baselines on the ScanNet and SUN RGB-D datasets, respectively. Furthermore, we provide a comprehensive analysis to explain why our approach works.Comment: I want to update this manuscrip

Locating Hazardous Chemical Leakage Source Based on Cooperative Moving and Fixing Sensors

In dealing with sudden hazardous chemical leakage accidents, the key to solving the evacuation and transfer of personnel and important property is to determine the location of the leakage source and the information of the source strength to gauge the scope of the impact of leakage. The particle swarm optimization algorithm with an adaptive mutation factor is applied to the inverse calculation of leakage source strength to obtain the leakage source information, and the leakage source location problem is transformed into an optimization problem. The mobile sensor is then introduced into the fixed sensor network. The mobile sensor moving strategy based on an extended Kalman filter is proposed. The estimated value of the previous moment and the current time are used to update the estimation of the state variable, and then the mobile strategy is planned. The interference of the random error of the optimization algorithm on the path planning of the mobile sensor is reduced by introducing the optimized result memory and, thus, location efficiency is improved. Simulation results showed that the proposed method, which combines mobile with fixed sensors, greatly expanded the monitoring function of the network, reduced the number of fixed sensors, and enhanced the positioning accuracy

Biomechanical Evaluation of the Cross-link Usage and Position in the Single and Multiple Segment Posterior Lumbar Interbody Fusion

Objective: Previous studies have neither explored the usage of cross-links nor investigated the optimal position of the cross-links in posterior lumbar interbody fusion (PLIF). This study evaluates biomechanical properties of cross-links in terms of different fixation segments and optimal position in single- and multi-segment posterior lumbar interbody fusion. Methods: Two finite element (FE) models of instrumented lumbosacral spine with single-(L4/5) and multi-segment (L3-S1) PLIF surgery were simulated. On the basis of the two models, the benefits of the usage of cross-links were assessed and compared with the status of no application of cross-links. Moreover, the effects of position of cross-links on multi-segment PLIF surgery were studied in Upper, Middle, and Lower positions. Results: No significant difference was found in the range of motion (ROM), intersegmental rotational angle (IRA) of adjacent segments, and intradiscal pressure (IDP) regardless of the usage of cross-links in the single-segment PLIF surgery, while the cross-link increased the maximum von Mises stress in the fixation (MSF) under the axial rotation (53.65 MPa vs 41.42 MPa). In the multi-segment PLIF surgery, the usage of cross-links showed anti-rotational advantages indicated by ROM (Without Cross-link 2.35o, Upper, 2.24o; Middle, 2.26o; Lower, 2.30o) and IRA (Without Cross-link 1.19o, Upper, 1.08o; Middle, 1.09o; Lower, 1.13o). The greatest values of MSF were found in without cross-link case under the flexion, lateral bending, and axial rotation (37.48, 62.61, and 86.73 MPa). The application of cross-links at the Middle and Lower positions had lower values of MSF (48.79 and 69.62 MPa) under the lateral bending and axial rotation, respectively. Conclusion: The application of cross-links was not beneficial for the single-segment PLIF, while it was found highly advantageous for the multi-segment PLIF. Moreover, the usage of cross-links at the Middle or Lower positions resulted in a better biomechanical stability.</p