Diesel degradation capability and environmental robustness of strain Pseudomonas aeruginosa WS02.

Petroleum hydrocarbon (PHC) degrading bacteria have been frequently discovered. However, in practical application, a single species of PHC degrading bacterium with weak competitiveness may face environmental pressure and competitive exclusion due to the interspecific competition between petroleum-degrading bacteria as well as indigenous microbiota in soil, leading to a reduced efficacy or even malfunction. In this study, the diesel degradation ability and environmental robustness of an endophytic strain Pseudomonas aeruginosa WS02, were investigated. The results show that the cell membrane surface of WS02 was highly hydrophobic, and the strain secreted glycolipid surfactants. Genetic analysis results revealed that WS02 contained multiple metabolic systems and PHC degradation-related genes, indicating that this strain theoretically possesses the capability of oxidizing both alkanes and aromatic hydrocarbons. Gene annotation also showed many targets which coded for heavy metal resistant and metal transporter proteins. The gene annotation-based inference was confirmed by the experimental results: GC-MS analysis revealed that short chain PHCs (C10-C14) were completely degraded, and the degradation of PHCs ranging from C15-C22 were above 90% after 14 d in diesel-exposed culture; Heavy metal (Mn2+, Pb2+ and Zn2+) exposure was found to affect the growth of WS02 to some extent, but not its ability to degrade diesel, and the degradation efficiency was still maintained at 39-59%. WS02 also showed a environmental robustness along with PHC-degradation performance in the co-culture system with other bacterial strains as well as in the co-cultured system with the indigenous microbiota in soil fluid extracted from a PHC-contaminated site. It can be concluded that the broad-spectrum diesel degradation efficacy and great environmental robustness give P. aeruginosa WS02 great potential for application in the remediation of PHC-contaminated soil.

Maximum surgical blood order schedule for flap reconstruction in oral and maxillofacial cancer patients.

BACKGROUND: We established a MSBOS for flap reconstruction in oral and maxillofacial cancer patients. METHOD: We enrolled 2080 cases of oral and maxillofacial flap reconstruction from January 1, 2010 to December 31, 2021. Patient data were collected, including age, sex, BMI, preoperative Hb levels, ASA grade, T stage, flap type, tumor location, and bone flap. Scoring criteria were established based on a multivariate model of independent risk variables and their odds ratios. Two flap-type groups were divided into low-risk, intermediate-risk and high-risk groups by the scoring criteria, and analyzed using univariate and multivariate logistic regression. Perioperative transfusion analysis identified independent risk factors at various Hb levels. The cumulative percentage of patients requiring perioperative blood transfusion for each surgical procedure was calculated to establish the MSBOS. RESULTS: (1) Regression analysis showed that BMI, tumor T staging, ASA grade, preoperative Hb level (male: Hb < 130 g/L, female: Hb < 120 g/L), and bone flap were independent risk factors for perioperative blood transfusion. (2) Regression analysis showed that independent risk factors for perioperative transfusion included the following: BMI, tumor T3-T4 stage, ASA III, IV grade, and free flap/pediculated flap/bone flap in patients with different Hb levels; T3-T4 stage, ASA grade III-IV in mildly anemic patients; and ASA grade III-IV in moderately anemic patients. (3) A MSBOS was established for flap reconstruction in head and neck cancer patients. CONCLUSION: A MSBOS for head and neck cancer procedures was reduced by approximately 30% perioperative blood preparation while ensuring that clinical blood use standards were met. It help optimize blood inventory, and save blood resources.

The temporal shift of peri-implant microbiota during the biofilm formation and maturation in a canine model.

OBJECTIVES: Although the mature peri-implant biofilm composition is well studied, there is very little information on the succession of in vivo dental implant colonization. The aim of this study was to characterize the temporal changes and diversity of peri-implant supra-mucosal and sub-mucosal microbiota during the process of the plaque maturation. MATERIALS AND METHODS: Dental implants (n = 25) were placed in the mandible of 3 beagle dogs. Illumina MiSeq sequencing of the hypervariable V3-V4 region of the 16S rRNA gene amplicons was used to characterize the supra/sub-mucosal microbiota in the peri-implant niches at 1day (T1), 7days (T2), 14days (T3), 21days (T4) and 28days (T5) after Phase Ⅱ surgery of the healing abutment placement. QIIME, Mothur, LEfSe and R-package were used for downstream analysis. RESULTS: A total of 1184 operational taxonomic units (OTUs), assigned into 22 phyla, 264 genera and 339 species were identified. In supra-mucosal niches, the alpha parameters of shannon, sobs and chao1 displayed significant differences between T1 and other time-points. However, in sub-mucosal niches, only sobs, chao1, and ace indexes displayed significant differences between T1 and T3, and T1 and T5. Beta-diversity showed statistically significant difference between T1 and T2, T3, T4, T5 within both sub-mucosal and supra-mucosal plaque. The phyla Bacteroidetes, Proteobacteria and Firmicutes were the most dominant phyla of both sub-mucosal and supra-mucosal niches at all time-points and Firmicutes increased during the maturation of peri-implant plaque. At the genus level, Neisseria decreased significantly after T1 suggesting the establishment of an anaerobic microenvironment. A decrease of Porphyromonas during the formation of sub-mucosal microbial community was also detected. Co-occurrence network analysis exhibited a more complicated co-occurrence relationship of bacterial species in the sub-mucosal niches. Fusobacterium nucleatum, Filifactor villosus, and some other species may play a crucial role in biofilm maturation. CONCLUSIONS: The present results suggested that the development of peri-implant biofilm followed a similar pattern to dental plaque formation. Sub-mucosal biofilm may go through a more complicated procedure of maturation than supra-mucosal biofilm.

A method for measuring banana pseudo-stem phenotypic parameters based on handheld mobile LiDAR and IMU fusion.

Diameter and height are crucial morphological parameters of banana pseudo-stems, serving as indicators of the plant's growth status. Currently, in densely cultivated banana plantations, there is a lack of applicable research methods for the scalable measurement of phenotypic parameters such as diameter and height of banana pseudo-stems. This paper introduces a handheld mobile LiDAR and Inertial Measurement Unit (IMU)-fused laser scanning system designed for measuring phenotypic parameters of banana pseudo-stems within banana orchards. To address the challenges posed by dense canopy cover in banana orchards, a distance-weighted feature extraction method is proposed. This method, coupled with Lidar-IMU integration, constructs a three-dimensional point cloud map of the banana plantation area. To overcome difficulties in segmenting individual banana plants in complex environments, a combined segmentation approach is proposed, involving Euclidean clustering, Kmeans clustering, and threshold segmentation. A sliding window recognition method is presented to determine the connection points between pseudo-stems and leaves, mitigating issues caused by crown closure and heavy leaf overlap. Experimental results in banana orchards demonstrate that, compared with manual measurements, the mean absolute errors and relative errors for banana pseudo-stem diameter and height are 0.2127 cm (4.06%) and 3.52 cm (1.91%), respectively. These findings indicate that the proposed method is suitable for scalable measurements of banana pseudo-stem diameter and height in complex, obscured environments, providing a rapid and accurate inter-orchard measurement approach for banana plantation managers.

Extracellular vesicle surface display of αPD-L1 and αCD3 antibodies via engineered late domain-based scaffold to activate T-cell anti-tumor immunity.

Extracellular vesicles (EVs) are emerging as promising carriers for the delivery of therapeutic biologics. Genetic engineering represents a robust strategy for loading proteins of interest into EVs. Identification of EV-enriched proteins facilitates protein cargo loading efficiency. Many EV-enriched proteins are sorted into EVs via an endosomal sorting complex required for transport (ESCRT)-dependent pathway. In parallel, viruses hijack this EV biosynthesis machinery via conserved late domain motifs to promote egress from host cells. Inspired by the similarity of biogenesis between EVs and viruses, we developed a synthetic, Late domain-based EV scaffold protein that enables the display of a set of single chain variable fragments (scFvs) on the EV surface. We named this scaffold the Late domain-based exosomal antibody surface display platform (LEAP). We applied the LEAP scaffold to reprogramme HEK293T cell-derived EVs to elicit T-cell anti-tumor immunity by simultaneously displaying αPD-L1 and αCD3 scFvs on the EV surface (denoted as αPD-L1×αCD3 bispecific T-cell engaging exosomes, BiTExos). We demonstrated that αPD-L1×αCD3 BiTExos actively redirected T cells to bind to PD-L1+ tumor cells, promoting T-cell activation, proliferation and tumoricidal cytokine production. Furthermore, the αPD-L1×αCD3 BiTExos promoted T-cell infiltration into the tumor microenvironment to mitigate the tumor burden in vivo. Our study suggested that the LEAP scaffold may serve as a platform for EV surface display and could be applied for a broad range of EV-based biomedical applications.

Changes in slow-wave sleep characteristics in Parkinson's disease patients with mild-moderate depression.

INTRODUCTION: Depression and sleep disturbances are commonly seen non-motor symptoms in patients with Parkinson's disease (PD). This study used polysomnography to examine the relationship between mild-moderate depression in PD and sleep characteristics, particularly slow wave activities (SWA). METHODS: 59 PD patients were split into two groups: nd-PD (n = 27) (patients with PD without depression) and d-PD (n = 32) (patients with PD with mild-moderate depression). Their clinical features, polysomnography parameters, and demographics were evaluated. Early and late sleep SWA spectrum densities and overnight SWA decline in different brain regions were particularly analyzed. RESULTS: Non-rapid eye movement 3 (N3) sleep duration and percentage were greater in the d-PD group. N3 percentage was linked to depression (p = 0.014). During late sleep, higher SWA (0.5-4Hz) in the frontal and central regions, higher low-SWA (0.5-2Hz) in the whole brain, central and occipital regions, and higher high-SWA (2-4Hz) in the frontal region was observed in the d-PD group. During early sleep, there was also higher low-SWA (0.5-2Hz) in the occipital region. Patients in d-PD group exhibited reduced overnight high-SWA (2-4Hz) decline (Δhigh-SWA) in the whole brain and occipital regions. Δhigh-SWA(2-4Hz) in the occipital region were associated with depression (p = 0.049). CONCLUSION: PD patients with mild-moderate depression have impaired slow wave sleep, exhibiting as increased N3 sleep, SWA, and reduced overnight SWA decline. This implies that synaptic strength reduction during sleep and impaired synaptic homeostasis regulation may be associated with depression in PD. Reduced overnight high-SWA decline in the occipital region may serve as a novel electrophysiological biomarker for indicating depression in PD.

A manganese-oxidizing bacterium-Enterobacter hormaechei strain DS02Eh01: Capabilities of Mn(II) immobilization, plant growth promotion and biofilm formation.

While biogenic Mn oxides (BioMnOx) generated by Mn(II)-oxidizing bacteria (MOB) have attracted increasing attention, a MOB strain isolated from Mn-polluted sediments was identified and assigned as Enterobacter hormaechei DS02Eh01. Its Mn(II) immobilization activity, plant growth-promoting traits, and biofilm formation capability were investigated. The results showed that strain DS02Eh01 was found to be able to tolerate Mn(II) up to 122 mM. The strain immobilized Mn(II) in aquatic media mainly through extracellular adsorption, bio-oxidation and pH-induced precipitation as well as manganese oxidation. DS02Eh01-derived BioMnOx are negatively charged and have a larger specific surface area (86.70 m2/g) compared to the previously reported BioMnOx. The strain can immobilize Mn(II) at extreme levels, for instance, when it was exposed to 20 mM Mn(II), about 59% of Mn(II) were found immobilized and 17% of Mn(II) were converted to MnOx. The SEM and TEM observation revealed that the DS02Eh01-derived BioMnOx were aggregates doped with granules and microbial pellets. The precipitated Mn(II) and the Mn(III)/Mn(IV) oxides co-existed in BioMnOx, in which Mn(II) and Mn(IV) were found dominant with Mn(II) accounting for 49.6% and Mn(IV) accounting for 41.3%. DS02Eh01 possesses plant growth-promoting traits and biofilm formation capacity even under Mn(II) exposure. Mn(II) exposure at 5 mM was found to stimulate strain DS02Eh01 to form biofilms, from which, the extracted EPS was mainly composed of aromatic proteins. This study reveals that E. hormaechei strain DS02Eh01 possesses the potential in environmental ecoremediation via coupling processes of macrophytes extraction, biochemical immobilization and biosorption.

Influence of implant protrusion length on non-grafting osteotome sinus floor elevation with simultaneous implant: a 3- to 9-year retrospective study.

BACKGROUND: This study analyzed the influence of implant protrusion length (IPL) on the possible factors that affect the long-term outcomes utilizing non-grafting osteotome sinus floor elevation (OSFE) with simultaneous implant placement, and to explore the optimal range of IPL. MATERIALS AND METHODS: A retrospective study design was adopted. The clinical and radiographic data of 105 implants in 65 patients were collected after 3-9 (mean 5.04) years follow-up. IPL was divided into three groups (group1, IPL<2mm; group2, 2mm≤IPL<4mm; group3, IPL≥4mm). Endo-sinus bone gain (ESBG), peri-implant marginal bone loss (MBL), bone to implant contact length (BICL), and percentage of ESBG (%ESBG) were used to evaluate non-grafting OSFE. A Kaplan-Meier analysis was performed to assess the cumulative survival rate. Multiple linear regression model was used to explore the relationship between the possible influence factors and ESBG. Analysis of variance (ANOVA) was applied to explore the correlation of IPL with ESBG, MBL, BICL, and %ESBG. RESULTS: A total of 102 implants in 62 patients fulfilled the survival criteria, giving the cumulative survival rates of 96.4% and 94.1% for implant-based analysis and patient-based analysis, respectively. The mean ESBG, MBL, and BICL at the latest follow-up were 1.95±0.88 mm, 0.58±0.68 mm, and 5.51±1.47 mm. ESBG was found to be positively correlated to IPL. A significant decreased bone formation efficiency was found when IPL was over 4 mm (P=0.02). CONCLUSIONS: An optimal range of IPL within 4 mm was recommended for better long-term outcomes when applying non-grafting OSFE with simultaneous implant placement.

Different Cell and Tissue Behavior of Micro-/Nano-Tubes and Micro-/Nano-Nets Topographies on Selective Laser Melting Titanium to Enhance Osseointegration.

BACKGROUND AND PURPOSE: Micro-/nano-tubes (TNTs) and micro-/nano-nets (TNNs) are the common and sensible choice in the first step of combined modifications of titanium surface for further functionalization in the purpose of extended indications and therapeutic effect. It is important to recognize the respective biologic reactions of these two substrates for guiding a biologically based first-step selection. MATERIALS AND METHODS: TNTs were produced by anodic oxidation and TNNs were formed by alkali-heat treatment. The original selective laser melting (SLM) titanium surface was set as control. Surface characterization was evaluated by scanning electron microscopy, surface roughness, and water contact angle measurements. Osteoclastogenesis and osteogenesis were measured. MC3T3-E1 cells and RAW 264.7 cells were used for in vitro assay in terms of adhesion, proliferation, and differentiation. In vivo assessments were taken on Beagle dogs with micro-CT and histological analysis. RESULTS: TNN and TNT groups performed decreased roughness and increased hydrophilicity compared with SLM group. For biological detections, the highest ALP activity and osteogenesis-related genes expression were observed in TNT group followed by TNN group (P <0.05). Interestingly, when it comes to the osteoclastogenesis, TNNs displayed lowest TRAP activity and osteoclastogenesis-related genes expression and TNTs were lower than SLM but higher than TNNs (P <0.05). BV/TV around implants was highest in TNT group after 4 weeks (P <0.05). HE, ALP and TRAP staining showed that osteogenic and osteoclastic activity around TNTs were both higher than TNNs (P <0.05). CONCLUSION: TNNs and TNTs have dual advantages in promotion of osteogenesis and inhibition of osteoclastogenesis. Furthermore, TNNs showed better capability in inhibiting osteoclast activity while TNTs facilitated stronger osteogenesis. Our results implied that TNT substrates would take advantage in early application after implantation, while diseases with inappropriate osteoclast activity would prefer TNN substrates, which will guide a biologically based first-step selection on combined modification for different clinical purposes.

Antibacterial Evaluation of Lithium-Loaded Nanofibrous Poly(L-Lactic Acid) Membranes Fabricated via an Electrospinning Strategy.

Lithium (Li) reportedly has anti-bacterial properties. Thus, it is an ideal option to modify barrier membranes used for guided bone regeneration to inhibit the bacterial adhesion. The aims of this study were to fabricate and characterize nanofibrous poly(L-lactic acid) (PLLA) membranes containing Li, and investigate their antibacterial effects on Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in vitro. Li (5%Li, 10%Li, and 15%Li)-loaded nanofibrous PLLA membranes were fabricated using an electrospinning technique, and characterized via scanning electron microscopy, X-ray photoelectron spectroscopy, X-ray diffraction, a contact angle measuring device, and a universal testing machine. Sustained release of Li ions was measured over a 14-day period and biocompatibility of the Li-PLLA membranes was investigated. Evaluation of bacterial adhesion and antibacterial activity were conducted by bacterial colony counting, LIVE/DEAD staining and inhibition zone method using P.gingivalis and A.actinomycetemcomitans. Of the three Li-loaded membranes assessed, the 10%Li-PLLA membrane had the best mechanical properties and biocompatibility. Adhesion of both P.gingivalis and A.actinomycetemcomitans on Li-PLLA membranes was significantly lower than adhesion on pure PLLA membranes, particularly with regard to the 10%Li and 15%Li membranes. Significant antibacterial activity of Li-PLLA were also observed against according to the inhibition zone test. Given their better mechanical properties, biocompatibility, and antibacterial activity, PLLAs with 10%Li are a better choice for future clinical utilization. The pronounced antibacterial effects of Li-loaded PLLA membranes sets the stage for further application in guided bone regeneration.