Integrated Biofilm Modification and Transcriptional Analysis for Improving Fengycin Production in Bacillus amyloliquefaciens.

Although fengycin exhibits broad-spectrum antifungal properties, its application is hindered due to its low biosynthesis level and the co-existence of iturin A and surfactin in Bacillus amyloliquefaciens HM618, a probiotic strain. In this study, transcriptome analysis and gene editing were used to explore the potential mechanisms regulating fengycin production in B. amyloliquefaciens. The fengycin level of B. amyloliquefacien HM-3 (∆itu-ΔsrfAA) was 88.41 mg/L after simultaneously inhibiting the biosyntheses of iturin A and surfactin. The knockout of gene eps associated with biofilm formation significantly increased the fengycin level of the strain HM618, whereas the fengycin level decreased 32.05% after knocking out sinI, a regulator of biofilm formation. Transcriptome analysis revealed that the differentially expressed genes, involved in pathways of amino acid and fatty acid syntheses, were significantly down-regulated in the recombinant strains, which is likely associated with a decrease of fengycin production. The knockout of gene comQXPA and subsequent transcriptome analysis revealed that the ComQXPA quorum sensing system played a positive regulatory role in fengycin production. Through targeted genetic modifications and fermentation optimization, the fengycin production of the engineered strain HM-12 (∆itu-ΔsrfAA-ΔyvbJ) in a 5-L fermenter reached 1.172 g/L, a 12.26-fold increase compared to the fengycin level in the strain HM-3 (∆itu-ΔsrfAA) in the Erlenmeyer flask. Taken together, these results reveal the underlying metabolic mechanisms associated with fengycin synthesis and provide a potential strategy for improving fengycin production in B. amyloliquefaciens.

An Engineered Microbial Consortium Provides Precursors for Fengycin Production by Bacillus subtilis.

Fengycin has great potential for applications in biological control because of its biosafety and degradability. In this study, the addition of exogenous precursors increased fengycin production by Bacillus subtilis. Corynebacterium glutamicum was engineered to produce high levels of precursors (Thr, Pro, Val, and Ile) to promote the biosynthesis of fengycin. Furthermore, recombinant C. glutamicum and Yarrowia lipolytica providing amino acid and fatty acid precursors were co-cultured to improve fengycin production by B. subtilis in a three-strain artificial consortium, in which fengycin production was 2100 mg·L-1. In addition, fengycin production by the consortium in a 5 L bioreactor reached 3290 mg·L-1. Fengycin had a significant antifungal effect on Rhizoctonia solani, which illustrates its potential as a food preservative. Taken together, this work provides a new strategy for improving fengycin production by a microbial consortium and metabolic engineering.

Long-term oncological outcomes of robotic versus laparoscopic gastrectomy for gastric cancer: multicentre cohort study.

BACKGROUND: The aim of this multicentre cohort study was to compare the long-term oncological outcomes of robotic gastrectomy (RG) and laparoscopic gastrectomy (LG) for patients with gastric cancer. METHODS: Patients with gastric cancer who underwent radical gastrectomy by robotic or laparoscopic approaches from 1 March 2010 to 31 December 2018 at 10 high-volume centres in China were selected from institutional databases. Patients receiving RG were matched 1 : 1 by propensity score with patients undergoing LG. The primary outcome was 3-year disease-free survival. Secondary outcomes were overall survival and disease recurrence. RESULTS: Some 2055 patients who underwent RG and 4309 patients who had LG were included. The propensity score-matched cohort comprised 2026 RGs and 2026 LGs. Median follow-up was 41 (i.q.r. 39-58) months for the RG group and 39 (38-56) months for the LG group. The 3-year disease-free survival rates were 80.8% in the RG group and 79.5% in the LG group (log rank P = 0.240; HR 0.92, 95% c.i. 0.80 to 1.06; P = 0.242). Three-year OS rates were 83.9 and 81.8% respectively (log rank P = 0.068; HR 0.87, 0.75 to 1.01; P = 0.068) and the cumulative incidence of recurrence over 3 years was 19.3% versus 20.8% (HR 0.95, 0.88 to 1.03; P = 0.219), with no difference between groups. CONCLUSION: RG and LG in patients with gastric cancer are associated with comparable disease-free and overall survival.

An Adaptive, Lightweight, Body-Powered System for Prosthetic Hands Equipped with a Selectively Lockable Differential Mechanism.

The need for more practical, robust, and affordable prosthetic hands for amputees has led to significant advancements in their functionality. However, the challenge of designing prostheses that balance dexterity, functionality, and affordability still persists. The lack of prosthetic designs that can efficiently address the needs of amputees for both heavy manual labor and social interactions, while also being lightweight and dexterous, is still evident. This paper presents a design for an anthropomorphic, adaptive, lightweight, body-powered prosthetic hand built for performing Activities of Daily Living using a selectively lockable differential mechanism. The proposed differential mechanism allows the users to flex and lock the prosthesis' fingers in a wide range of poses and grasps. The performance of the body-powered prosthesis is experimentally validated with three different types of experiments: i) object grasping, ii) total grasping strength, and iii) individual finger force exertion. The resultant prosthetic hand is lightweight and comfortable to wear and is adequately capable of grasping a wide range of objects, including items commonly used in cleaning and cooking applications where waterproofness is essential.

Enhancing fengycin production in the co-culture of Bacillus subtilis and Corynebacterium glutamicum by engineering proline transporter.

Fengycin possesses antifungal activity but has limited application due to its low yields. Amino acid precursors play a crucial role in fengycin synthesis. Herein, the overexpression of alanine, isoleucine, and threonine transporter-related genes in Bacillus subtilis increased fengycin production by 34.06%, 46.66%, and 7.83%, respectively. Particularly, fengycin production in B. subtilis reached 871.86 mg/L with the addition of 8.0 g/L exogenous proline after enhancing the expression of the proline transport-related gene opuE. To overcome the metabolic burden caused by excessive enhancement of gene expression for supplying precursors, B. subtilis and Corynebacterium glutamicum which produced proline, were co-cultured, which further improved fengycin production. Fengycin production in the co-culture of B. subtilis and C. glutamicum in shake flasks reached 1554.74 mg/L after optimizing the inoculation time and ratio. The fengycin level in the fed-batch co-culture was 2309.96 mg/L in a 5.0-L bioreactor. These findings provide a new strategy for improving fengycin production.

Achieving ultrastability and efficient lithium storage capacity with high-energy iron(II) oxalate anode materials by compositing Ge nano-conductive sites.

Transition metal oxalates (TMOxs, represented by iron oxalate) have attracted considerable interest in anode materials due to their excellent lithium storage properties and consistent cyclic performance. Although investigations into their electrochemical capabilities and lithium storage mechanisms are gradually deepening, the complex and varied electrochemical reactions in the initial cycle, poor inherent conductivity, and high irreversible capacity constrain their further development. Herein, to solve the above-mentioned problems, we controlled the hydrothermal synthesis conditions of iron oxalate with the assistance of organic solvents, which induced the growth of iron oxalate crystals with nano Ge metal as the core. The metal Ge space sites compounded to the stacked iron oxalate particles act as conductive nodes and metal frames, which enhances both the strength of iron oxalate samples and electronic conductivity and lithium-ion diffusion inside the electrode materials. This special structure enhances the electrochemical activity of iron oxalates and improves their lithium storage capability. The iron oxalate @ nano Ge metal composite (FCO@Ge-1) exhibits an excellent cycling performance and an appreciable reversible specific capacity (1090 mA h g-1 after 200 cycles at 1 A g-1). The obvious polarization and variation of the electrochemical reaction in the initial cycle of iron oxalate are reduced by compositing nano Ge metal. It is demonstrated that nano Ge metal can promote reversible capacity retention from 67.72% to 80.69% in the early cycles. The distinctive structure of iron oxalate @ nano Ge metal composite provides a fresh pathway to enhance oxalate electrochemical reversible lithium storage activity and develop high-energy electrode material by constructing composite space conductive sites.

Robotic Gastrectomy Versus Laparoscopic Gastrectomy for Gastric Cancer: A Multicenter Cohort Study of 5402 Patients in China.

OBJECTIVE: A large-scale multicenter retrospective cohort study was conducted to compare the short- and long-term outcomes of robotic gastrectomy (RG) and laparoscopic gastrectomy (LG) for gastric cancer. SUMMARY OF BACKGROUND DATA: RG is being increasingly used worldwide, but data from large-scale multicenter studies on the short- and long-term oncologic outcomes of RG versus LG are limited. The potential benefits of RG compared with LG for gastric cancer remain controversial. METHODS: Data from eligible patients who underwent RG or LG for gastric cancer of 11 experienced surgeons from 7 centers in China between March 2010 and October 2019 were collected. The RG group was matched 1:1 with the LG group by using propensity score matching. The primary outcome was postoperative complications. RESULTS: After propensity score matching, a well-balanced cohort of 3552 patients was included for further analysis. The occurrence of overall complications (12.6% vs 15.2%, P = 0.023) was lower in the RG group than in the LG group. RG was associated with less blood loss (126.8 vs 142.5 mL, P < 0.001) and more retrieved lymph nodes in total (32.5 vs 30.7, P < 0.001) and in suprapancreatic areas (13.3 vs 11.6, P < 0.001).The long-term oncological outcomes were comparable between the two groups. CONCLUSIONS: The results of this multicenter study demonstrate that RG is a safe and effective treatment for gastric cancer when performed by experienced surgeons, although longer operation time and higher costs are still concerns about RG. This study provides evidence suggesting that RG may represent an alternative surgical treatment to LG.

Mutualistic microbial community of Bacillus amyloliquefaciens and recombinant Yarrowia lipolytica co-produced lipopeptides and fatty acids from food waste.

Bioconversion is an important method for transforming food waste (FW) into high value-added products, rendering it harmless, and recycling resources. An artificial microbial consortium (AMC) was constructed to produce FW-based lipopeptides in order to investigate the strategy of FW bioconversion into value-added products. Exogenous fatty acids as a precursor significantly improved the lipopeptide production of Bacillus amyloliquefaciens HM618. To enhance fatty acid synthesis and efflux in AMC, the recombinant Yarrowia lipolytica YL21 (strain YL21) was constructed by screening 12 target genes related to fatty acids to replace exogenous fatty acids in order to improve lipopeptide production. The levels of fengycin, surfactin, and iturin A in the AMC of strains HM618 and YL21 reached 76.19, 192.80, and 31.32 mg L-1, increasing 7.24-, 12.13-, and 3.23-fold compared to the results from the pure culture of strain HM618 in flask with Landy medium, respectively. Furthermore, free fatty acids were almost undetectable in the co-culture of strains HM618 and YL21, although its level was around 1.25 g L-1 in the pure culture of strain YL21 with Landy medium. Interestingly, 470.24 mg L-1 of lipopeptides and 18.11 g L-1 of fatty acids were co-produced in this AMC in a bioreactor with FW medium. To our knowledge, it is the first report of FW biotransformation into co-produce of lipopeptides and fatty acids in the AMC of B. amyloliquefaciens and Y. lipolytica. These results provide new insights into the biotransformation potential of FW for value-added co-products by AMC.

Improved Production of Fengycin in Bacillus subtilis by Integrated Strain Engineering Strategy.

Fengycin is a lipopeptide with broad-spectrum antifungal activity. However, its low yield limits its commercial application. Therefore, we iteratively edited multiple target genes associated with fengycin synthesis by combinatorial metabolic engineering. The ability of Bacillus subtilis 168 to manufacture lipopeptides was restored, and the fengycin titer was 1.81 mg/L. Fengycin production was further increased to 174.63 mg/L after knocking out pathways associated with surfactin and bacillaene synthesis and replacing the native promoter (PppsA) with the Pveg promoter. Subsequently, fengycin levels were elevated to 258.52 mg/L by upregulating the expression of relevant genes involved in the fatty acid pathway. After blocking spore and biofilm formation, fengycin production reached 302.51 mg/L. Finally, fengycin production was increased to approximately 885.37 mg/L after adding threonine in the optimized culture medium, which was 488-fold higher compared with that of the initial strain. Integrated strain engineering provides a strategy to construct a system for improving fengycin production.

[Influence of COVID-19 Prevention and Control Measures on PM2.5 Concentration, Particle Size Distribution, Chemical Composition, and Source in Zhengzhou, China].

The COVID-19 lockdown was a typical occurrence of extreme emission reduction, which presented an opportunity to study the influence of control measures on particulate matter. Observations were conducted from January 16 to 31, 2020 using online observation instruments to investigate the characteristics of PM2.5 concentration, particle size distribution, chemical composition, source, and transport before (January 16-23, 2020) and during (January 24-31, 2020) the COVID-19 lockdown in Zhengzhou. The results showed that the atmospheric PM2.5 concentration decreased by 4.8% during the control period compared with that before the control in Zhengzhou. The particle size distribution characteristics indicated that there was a significant decrease in the mass concentration and number concentration of particles in the size range of 0.06 to 1.6 µm during the control period. The chemical composition characteristics of PM2.5 showed that secondary inorganic ions (sulfate, nitrate, and ammonium) were the dominant component of PM2.5, and the significant increase in PM2.5 was mainly owing to the decrease in NO3- concentration during the control period. The main sources of PM2.5 identified by the positive matrix factorization (PMF) model were secondary sources, combustion sources, vehicle sources, industrial sources, and dust sources. The emissions from vehicle sources, industrial sources, and dust sources decreased significantly during the control period. The results of analyses using the backward trajectory method and potential source contribution factor method indicated that the effects of transport from surrounding areas on PM2.5 concentration decreased during the control period. In summary, vehicle and industrial sources should be continuously controlled, and regional combined prevention and control should be strengthened in the future in Zhengzhou.

Facile Dissolution-Crystallization Strategy to Achieve Rapid and Uniform Distribution of Sulfur on Porous Carbon for Lithium-Sulfur Batteries.

In this work, we proposed a facile dissolution-crystallization strategy based on density functional theory calculations to achieve rapid as well as uniform distribution of sulfur on porous carbon. Sulfur-containing solution can completely penetrate porous material and in preference remove into the pores under the influence of capillary force, and sulfur tends to crystallize on the defective even non-defective carbon matrix rather than agglomerate. The S/PC composites prepared by this method can still achieve uniform distribution of sulfur when the sulfur content is as high as 85%. All operations can be completed within a few minutes without any heating. Compared with common melt-diffusion and vapor-phase infusion, this approach has lower energy consumption and is simple, safe, continuous, and rapid.

An Accessible, Open-Source Dexterity Test: Evaluating the Grasping and Dexterous Manipulation Capabilities of Humans and Robots.

Evaluating the dexterity of human and robotic hands through appropriate benchmarks, scores, and metrics is of paramount importance for determining how skillful humans are and for designing and developing new bioinspired or even biomimetic end-effectors (e.g., robotic grippers and hands). Dexterity tests have been used in industrial and medical settings to assess how dexterous the hands of workers and surgeons are as well as in robotic rehabilitation settings to determine the improvement or deterioration of the hand function after a stroke or a surgery. In robotics, having a comprehensive dexterity test can allow us to evaluate and compare grippers and hands irrespectively of their design characteristics. However, there is a lack of well defined metrics, benchmarks, and tests that quantify robot dexterity. Previous work has focused on a number of widely accepted functional tests that are used for the evaluation of manual dexterity and human hand function improvement post injury. Each of these tests focuses on a different set of specific tasks and objects. Deriving from these tests, this work proposes a new modular, affordable, accessible, open-source dexterity test for both humans and robots. This test evaluates the grasping and manipulation capabilities by combining the features and best practices of the aforementioned tests, as well as new task categories specifically designed to evaluate dexterous manipulation capabilities. The dexterity test and the accompanying benchmarks allow us to determine the overall hand function recovery and dexterity of robotic end-effectors with ease. More precisely, a dexterity score that ranges from 0 (simplistic, non-dexterous system) to 1 (human-like system) is calculated using the weighted sum of the accuracy and task execution speed subscores. It should also be noted that the dexterity of a robotic system can be evaluated assessing the efficiency of either the robotic hardware, or the robotic perception system, or both. The test and the benchmarks proposed in the study have been validated using extensive human and robot trials. The human trials have been used to determine the baseline scores for the evaluation system. The results show that the time required to complete the tasks reduces significantly with trials indicating a clear learning curve in mastering the dexterous manipulation capabilities associated with the imposed tasks. Finally, the time required to complete the tasks with restricted tactile feedback is significantly higher indicating its importance.

An Adaptive, Affordable, Open-Source Robotic Hand for Deaf and Deaf-Blind Communication Using Tactile American Sign Language.

Currently, ~ 1.5 million American deaf-blind individuals depend on the availability of interpreting services to communicate in their primary conversational language, tactile American Sign Language (ASL). In an effort to give the deaf-blind community access to a device that facilitates independent communication using tactile ASL, we developed TATUM (Tactile ASL Translational User Mechanism). TATUM employs 15 degrees of actuation in a hand-wrist system that is capable of signing the 26-letter ASL alphabet. Leveraging Interpres, an independent cloud-based service, all servo sequences that render desired fingerspelled letters and ASL words are stored in a web application programming interface (API). A validation study including both deaf and deaf-blind participants confirmed that the TATUM hand mimics a human hand both in size and feel. The current design of TATUM attained an average recognition rate of 94.7% in visual validation, indicative of the potential to support deaf and deaf-blind individuals in communicating via visual and tactile ASL.

On Differential Mechanisms for Underactuated, Lightweight, Adaptive Prosthetic Hands.

Over the last decade underactuated, adaptive robot grippers and hands have received an increased interest from the robotics research community. This class of robotic end-effectors can be used in many different fields and scenarios with a very promising application being the development of prosthetic devices. Their suitability for the development of such devices is attributed to the utilization of underactuation that provides increased functionality and dexterity with reduced weight, cost, and control complexity. The most critical components of underactuated, adaptive hands that allow them to perform a broad set of grasp poses are appropriate differential mechanisms that facilitate the actuation of multiple degrees of freedom using a single motor. In this work, we focus on the design, analysis, and experimental validation of a four output geared differential, a series elastic differential, and a whiffletree differential that can incorporate a series of manual and automated locking mechanisms. The locking mechanisms have been developed so as to enhance the control of the differential outputs, allowing for efficient grasp selection with a minimal set of actuators. The differential mechanisms are applied to prosthetic hands, comparing them and describing the benefits and the disadvantages of each.

Robotic- vs laparoscopic-assisted proctectomy for locally advanced rectal cancer based on propensity score matching: Short-term outcomes at a colorectal center in China.

BACKGROUND: Reports in the field of robotic surgery for rectal cancer are increasing year by year. However, most of these studies enroll patients at a relatively early stage and have small sample sizes. In fact, studies only on patients with locally advanced rectal cancer (LARC) and with relatively large sample sizes are lacking. AIM: To investigate whether the short-term outcomes differed between robotic-assisted proctectomy (RAP) and laparoscopic-assisted proctectomy (LAP) for LARC. METHODS: The clinicopathological data of patients with LARC who underwent robotic- or laparoscopic-assisted radical surgery between January 2015 and October 2019 were collected retrospectively. To reduce patient selection bias, we used the clinical baseline characteristics of the two groups of patients as covariates for propensity-score matching (PSM) analysis. Short-term outcomes were compared between the two groups. RESULTS: The clinical features were well matched in the PSM cohort. Compared with the LAP group, the RAP group had less intraoperative blood loss, lower volume of pelvic cavity drainage, less time to remove the pelvic drainage tube and urinary catheter, longer distal resection margin and lower rates of conversion (P < 0.05). However, the time to recover bowel function, the harvested lymph nodes, the postoperative length of hospital stay, and the rate of unplanned readmission within 30 days postoperatively showed no difference between the two groups (P > 0.05). The rates of total complications and all individual complications were similar between the RAP and LAP groups (P > 0.05). CONCLUSION: This retrospective study indicated that RAP is a safe and feasible method for LARC with better short-term outcomes than LAP, but we have to admit that the clinically significant of part of indicators are relatively small in the practical situation.

Adaptive, Tendon-Driven, Affordable Prostheses for Partial Hand Amputations: On Body-Powered and Motor Driven Implementations.

Adaptive, tendon-driven and affordable prosthetic devices have received an increased interest over the last decades. Prosthetic devices range from body-powered solutions to fully actuated systems. Despite the significant progress in the field, most existing solutions are expensive, heavy, and bulky, or they cannot be used for partial hand amputations. In this paper, we focus on the development of adaptive, tendon-driven, glove-based, affordable prostheses for partial hand amputations and we propose two compact and lightweight devices (a body powered and a motor driven version). The efficiency of the devices is experimentally validated and their performance is evaluated using two different types of tests: i) grasping tests that involve different everyday objects and ii) tests that assess the force exertion capabilities of the proposed prostheses.

Functional kinomics establishes a critical node of volume-sensitive cation-Cl- cotransporter regulation in the mammalian brain.

Cell volume homeostasis requires the dynamically regulated transport of ions across the plasmalemma. While the ensemble of ion transport proteins involved in cell volume regulation is well established, the molecular coordinators of their activities remain poorly characterized. We utilized a functional kinomics approach including a kinome-wide siRNA-phosphoproteomic screen, a high-content kinase inhibitor screen, and a kinase trapping-Orbitrap mass spectroscopy screen to systematically identify essential kinase regulators of KCC3 Thr991/Thr1048 phosphorylation - a key signaling event in cell swelling-induced regulatory volume decrease (RVD). In the mammalian brain, we found the Cl--sensitive WNK3-SPAK kinase complex, required for cell shrinkage-induced regulatory volume decrease (RVI) via the stimulatory phosphorylation of NKCC1 (Thr203/Thr207/Thr212), is also essential for the inhibitory phosphorylation of KCC3 (Thr991/Thr1048). This is mediated in vivo by an interaction between the CCT domain in SPAK and RFXV/I domains in WNK3 and NKCC1/KCC3. Accordingly, genetic or pharmacologic WNK3-SPAK inhibition prevents cell swelling in response to osmotic stress and ameliorates post-ischemic brain swelling through a simultaneous inhibition of NKCC1-mediated Cl- uptake and stimulation of KCC3-mediated Cl- extrusion. We conclude that WNK3-SPAK is an integral component of the long-sought "Cl-/volume-sensitive kinase" of the cation-Cl- cotransporters, and functions as a molecular rheostat of cell volume in the mammalian brain.

Comprehensive identification of host modulators of HIV-1 replication using multiple orthologous RNAi reagents.

RNAi screens have implicated hundreds of host proteins as HIV-1 dependency factors (HDFs). While informative, these early studies overlap poorly due to false positives and false negatives. To ameliorate these issues, we combined information from the existing HDF screens together with new screens performed with multiple orthologous RNAi reagents (MORR). In addition to being traditionally validated, the MORR screens and the historical HDF screens were quantitatively integrated by the adaptation of an established analysis program, RIGER, for the collective interpretation of each gene's phenotypic significance. False positives were addressed by the removal of poorly expressed candidates through gene expression filtering, as well as with GESS, which identifies off-target effects. This workflow produced a quantitatively integrated network of genes that modulate HIV-1 replication. We further investigated the roles of GOLGI49, SEC13, and COG in HIV-1 replication. Collectively, the MORR-RIGER method minimized the caveats of RNAi screening and improved our understanding of HIV-1-host cell interactions.

Protein interaction discovery using parallel analysis of translated ORFs (PLATO).

Identifying physical interactions between proteins and other molecules is a critical aspect of biological analysis. Here we describe PLATO, an in vitro method for mapping such interactions by affinity enrichment of a library of full-length open reading frames displayed on ribosomes, followed by massively parallel analysis using DNA sequencing. We demonstrate the broad utility of the method for human proteins by identifying known and previously unidentified interacting partners of LYN kinase, patient autoantibodies, and the small-molecules gefitinib and dasatinib.

Systematic identification of synergistic drug pairs targeting HIV.

The systematic identification of effective drug combinations has been hindered by the unavailability of methods that can explore the large combinatorial search space of drug interactions. Here we present multiplex screening for interacting compounds (MuSIC), which expedites the comprehensive assessment of pairwise compound interactions. We examined ∼500,000 drug pairs from 1,000 US Food and Drug Administration (FDA)-approved or clinically tested drugs and identified drugs that synergize to inhibit HIV replication. Our analysis reveals an enrichment of anti-inflammatory drugs in drug combinations that synergize against HIV. As inflammation accompanies HIV infection, these findings indicate that inhibiting inflammation could curb HIV propagation. Multiple drug pairs identified in this study, including various glucocorticoids and nitazoxanide (NTZ), synergize by targeting different steps in the HIV life cycle. MuSIC can be applied to a wide variety of disease-relevant screens to facilitate efficient identification of compound combinations.