A novel thermophilic strain of Bacillus subtilis with antimicrobial activity and its potential application in solid-state fermentation of soybean meal.

Soybean meal (SBM) is the most important source of plant protein in animal feeds, containing around 41%-48% crude protein. Nevertheless, 70%-80% of these proteins is allergenic antigens that can have adverse implications for the gastrointestinal well-being of animals, especially to young animals. Microbial fermentation is one of the most cost-effective strategies used to reduce allergenic antigens from plant sources. In this study, we report the isolation and characterization of a novel probiotic Bacillus subtilis "L5" strain from lake mud. L5 demonstrated remarkable temperature tolerance across a broad temperature spectrum, thriving at 25°C, 37°C, and 50°C. In addition, antimicrobial assay revealed that L5 exhibits strong antimicrobial activity against Escherichia coli, effectively reducing or eliminating the growth of Gram-negative bacteria in SBM when fermented with L5. When applied to SBM fermentation, L5 efficiently reduced SBM antinutritional factors such as glycinin, ß-conglycinin, trypsin inhibitor, phytic acid, neutral detergent fiber, and acid detergent fiber, which in turn results in an increase in crude protein content and the free amino acid concentration. Our findings on the probiotic and fermentation capabilities of L5 suggest that this novel bacterium has dual functions that make it a strong candidate for improving the nutrient values of feed via its role in fermentation.IMPORTANCESoybean meal (SBM), containing 41%-48% crude protein, is the most important source of plant protein in animal feeds. Unfortunately, 70%-80% of the proteins in SBM is allergenic antigens including trypsin inhibition, ß-conglycinin, and conglycinin, which negatively affect intestine health and function. Microbial solid-state fermentation methods have been applied to animal feeds for decades, to eliminate antinutritional factors. Here, a novel potential probiotic Bacillus subtilis "L5" strain with high enzymatic activity and antimicrobial activity will be a great help to improve the quality and reproducibility of SBM fermentation.

Injectable, Antioxidative, and Tissue-Adhesive Nanocomposite Hydrogel as a Potential Treatment for Inner Retina Injuries.

Reactive oxygen species (ROS) have been recognized as prevalent contributors to the development of inner retinal injuries including optic neuropathies such as glaucoma, non-arteritic anterior ischemic optic neuropathy, traumatic optic neuropathy, and Leber hereditary optic neuropathy, among others. This underscores the pivotal significance of oxidative stress in the damage inflicted upon retinal tissue. To combat ROS-related challenges, this study focuses on creating an injectable and tissue-adhesive hydrogel with tailored antioxidant properties for retinal applications. GelCA, a gelatin-modified hydrogel with photo-crosslinkable and injectable properties, is developed. To enhance its antioxidant capabilities, curcumin-loaded polydopamine nanoparticles (Cur@PDA NPs) are incorporated into the GelCA matrix, resulting in a multifunctional nanocomposite hydrogel referred to as Cur@PDA@GelCA. This hydrogel exhibits excellent biocompatibility in both in vitro and in vivo assessments, along with enhanced tissue adhesion facilitated by NPs in an in vivo model. Importantly, Cur@PDA@GelCA demonstrates the potential to mitigate oxidative stress when administered via intravitreal injection in retinal injury models such as the optic nerve crush model. These findings underscore its promise in advancing retinal tissue engineering and providing an innovative strategy for acute neuroprotection in the context of inner retinal injuries.

Biophysical Electrical and Mechanical Stimulations for Promoting Chondrogenesis of Stem Cells on PEDOT:PSS Conductive Polymer Scaffolds.

The investigation of the effects of electrical and mechanical stimulations on chondrogenesis in tissue engineering scaffolds is essential for realizing successful cartilage repair and regeneration. The aim of articular cartilage tissue engineering is to enhance the function of damaged or diseased articular cartilage, which has limited regenerative capacity. Studies have shown that electrical stimulation (ES) promotes mesenchymal stem cell (MSC) chondrogenesis, while mechanical stimulation (MS) enhances the chondrogenic differentiation capacity of MSCs. Therefore, understanding the impact of these stimuli on chondrogenesis is crucial for researchers to develop more effective tissue engineering strategies for cartilage repair and regeneration. This study focuses on the preparation of poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) conductive polymer (CP) scaffolds using the freeze-drying method. The scaffolds were fabricated with varying concentrations (0, 1, 3, and 10 wt %) of (3-glycidyloxypropyl) trimethoxysilane (GOPS) as a crosslinker and an additive to tailor the scaffold properties. To gain a comprehensive understanding of the material characteristics and the phase aggregation phenomenon of PEDOT:PSS scaffolds, the researchers performed theoretical calculations of solubility parameters and surface energies of PSS, PSS-GOPS, and PEDOT polymers, as well as conducted material analyses. Additionally, the study investigated the potential of promoting chondrogenic differentiation of human adipose stem cells by applying external ES or MS on a PEDOT:PSS CP scaffold. Compared to the group without stimulation, the group that underwent stimulation exhibited significantly up-regulated expression levels of chondrogenic characteristic genes, such as SOX9 and COL2A1. Moreover, the immunofluorescence staining images exhibited a more vigorous fluorescence intensity of SOX9 and COL II proteins that was consistent with the trend of the gene expression results. In the MS experiment, the strain excitation exerted on the scaffold was simulated and transformed into stress. The simulated stress response showed that the peak gradually decreased with time and approached a constant value, with the negative value of stress representing the generation of tensile stress. This stress response quantification could aid researchers in determining specific MS conditions for various materials in tissue engineering, and the applied stress conditions could be further optimized. Overall, these findings are significant contributions to future research on cartilage repair and biophysical ES/MS in tissue engineering.

Isolation and characterization of a novel hydrolase-producing probiotic Bacillus licheniformis and its application in the fermentation of soybean meal.

Soybean meal (SBM) is one of the most important sources of plant-based protein in the livestock and poultry industry. However, SBM contains anti-nutritional factors (ANFs) such as glycinin, ß-conglycinin, trypsin inhibitor and phytic acid that can damage the intestinal health of animals, inevitably reducing growth performance. Fermentation using microorganisms with probiotic potential is a viable strategy to reduce ANFs and enhance the nutritional value of SBM. In this study, a novel potential probiotic Bacillus licheniformis (B4) with phytase, protease, cellulase and xylanase activity was isolated from camel feces. The ability of B4 to tolerate different pH, bile salts concentrations and temperatures were tested using metabolic activity assay. It was found that B4 can survive at pH 3.0, or 1.0% bile salts for 5 h, and displayed high proliferative activity when cultured at 50°C. Furthermore, B4 was capable of degrading glycinin, ß-conglycinin and trypsin inhibitor which in turn resulted in significant increases of the degree of protein hydrolysis from 15.9% to 25.5% (p < 0.01) and crude protein from 44.8% to 54.3% (p < 0.001). After fermentation with B4 for 24 h, phytic acid in SBM was reduced by 73.3% (p < 0.001), the neutral detergent fiber (NDF) and the acid detergent fiber of the fermented SBM were significantly decreased by 38.40% (p < 0.001) and 30.20% (p < 0.05), compared to the unfermented SBM sample. Our results suggested that the effect of solid-state fermented SBM using this novel B. licheniformis (B4) strain, could significantly reduce phytic acid concentrations whilst improving the nutritional value of SBM, presenting itself as a promising alternative to phytase additives.

Serum Osteopontin Level Is Positively Associated with Aortic Stiffness in Patients with Peritoneal Dialysis.

Background: Osteopontin (OPN) is regarded as a proinflammatory and proatherogenic molecule related to atherosclerosis. We aimed to evaluate the relationship between serum OPN and aortic stiffness (AS) of peritoneal dialysis (PD) patients. Methods: OPN and carotid-femoral pulse wave velocity (cfPWV) were measured by a commercial enzyme-linked immunosorbent assay kit and a validated tonometry system, respectively. Patients with cfPWV > 10 m/s were designated into the AS group. Results: Twenty-two patients (31.4%) were segregated into the AS group. Multivariate linear and logistic regression analysis showed that OPN was significantly related to cfPWV and was an independent predictor of AS. The receiver operating characteristic curve analysis showed that OPN was correlated with AS with an area under the curve of 0.903 (95% CI 0.809−0.961, p < 0.001). Conclusions: For PD patients, the serum OPN level was correlated with cfPWV and could play an important role in the process of AS.

Combination therapy of sorafenib and drug-eluting bead transarterial chemoembolization for advanced hepatocellular carcinoma with and without hepatic arteriovenous shunt.

BACKGROUND: To compare the efficacy and safety of combination therapy with sorafenib and drug-eluting bead transarterial chemoembolization (DEB-TACE) in advanced hepatocellular carcinoma (HCC) with or without hepatic arteriovenous shunt (HAVS). METHODS: This retrospective, single-center study enrolled 59 advanced HCC patients treated with combination therapy, of whom 33 (55.9%) patients had HAVS. Tumor response according to the mRECIST criteria was evaluated based on the CT images 1 month after TACE, and changes in the arterial enhancement ratio (AER) of tumors and portal vein tumor thrombosis were also documented. Time-to-progression (TTP), overall survival (OS), and prognostic factors were analyzed. Safety was evaluated with the incidence of TACE-related complications within 6 weeks after TACE. RESULTS: The tumor response between the two groups showed no significant difference in the objective response rate (69.2% in the group without HAVS vs 60.6% in the group with HAVS, p = 0.492) or disease control rate (92.3% vs 87.9%, p = 0.685). The two groups showed comparable TTP (4.23 vs 2.33 months, p = 0.235) and OS (12.77 vs 12.97 months, p = 0.910). A drop in the AER of tumors of more than 20% on post-TACE CT independently predicted better OS. With regard to safety, there was no significant difference between the two groups. CONCLUSION: For advanced HCC, combination therapy had equal efficacy and safety in patients with HAVS compared to those without HAVS, indicating that DEB-TACE is an optional and effective treatment in these patients.

Different FDG-PET metabolic patterns of anti-AMPAR and anti-NMDAR encephalitis: Case report and literature review.

INTRODUCTION: 18 F-fluorodeoxyglucose (FDG)-PET metabolic patterns of brain differ among autoimmune encephalitis with different neuronal surface antigens. In this case report, we compared the topographical relationship of cerebral glucose metabolism and antigen distribution in the patients with anti-NMDAR and anti-AMPAR encephalitis. Literature review summarized the common features of brain metabolism of autoimmune encephalitis. METHODS: The cerebral glucose metabolism was evaluated by FDG-PET/CT during acute-to-subacute stage of autoimmune encephalitis and after treatment. The stereo and quantitative analysis of cerebral metabolism used standardized z-score and visualized on three-dimensional stereotactic surface projection. To map NMDAR and AMPAR in human brain, we adopted genetic atlases from the Allen Institute and protein atlases from Zilles's receptor densities. RESULTS: The three-dimensional stereotactic surface projection displayed frontal-dominant hypometabolism in a 66-year-old female patient with anti-AMPAR encephalitis and occipital-dominant hypometabolism in a 29-year-old female patient with anti-NMDAR encephalitis. Receptor density maps revealed opposite frontal-occipital gradients of AMPAR and NMDAR, which reflect reduced metabolism in the correspondent encephalitis. FDG-PET hypometabolic areas possibly represent receptor hypofunction with spatial correspondence to receptor distributions of the autoimmune encephalitis. The reversibility of hypometabolism was in line with patients' cognitive improvement. The literature review summarized six features of metabolic anomalies of autoimmune encephalitis: (a) temporal hypermetabolism, (b) frontal hypermetabolism and (c) occipital hypometabolism in anti-NMDAR encephalitis, (d) hypometabolism in association cortices, (e) sparing of unimodal primary motor cortex, and (e) reversibility in recovery. CONCLUSIONS: The distinct cerebral hypometabolic patterns of autoimmune encephalitis were representative for receptor hypofunction and topographical distribution of antigenic receptors. The reversibility of hypometabolism marked the clinical recovery of autoimmune encephalitis and made FDG-PET of brain a valuable diagnostic tool.

Predictive imaging for tumor response to drug-eluting microsphere transarterial chemoembolization in patients with BCLC-C advanced hepatocellular carcinoma.

Drug-eluting microsphere transarterial chemoembolization (DEM-TACE) has been introduced to ensure more sustained and tumor-selective drug delivery for permanent embolization of HCC. The aim of this study was to determine the imaging characteristics that related to favourable treatment response in BCLC-C HCC patients treated with DEM-TACE. In total, 64 patients with BCLC-C HCC that treated with DEM-TACE using doxorubicin-eluted microspheres were retrospectively included. The images were assessed at baseline and at 4-12 weeks follow-up after receiving DEM-TACE. Pre- and post-procedural imaging characteristics were analysed by two independent radiologists and treatment response was evaluated using the modified Response Evaluation Criteria in Solid Tumors criteria. Multivariate analysis showed that vascular lake phenomenon (OR = 5.94, p = 0.03*), and homogeneous tumor enhancement (HTE) on cone-beam computed tomography (CBCT) during angiography (OR = 11.66, p < 0.001*) are associated with better radiological response. In contrast, residual tumor blush (OR = 0.11, p < 0.001*) is associated with worse radiological response. In conclusion, the initial tumor burden <50% (p = 0.012*) and HTE on CBCT (p = 0.040*) are good predictors for locoregional tumor control in patients with advanced HCCs, which can potentially improve patients' outcome.

Differential efficacy of genetically swapping GAL4.

Several large or mid-scale collections of Drosophila enhancer traps have been recently created to allow for genetic swapping of GAL4 coding sequences to versatile transcription activators or suppressors such as LexA, QF, split-GAL4 (GAL4-AD and GAL4-DBD), GAL80 and QS. Yet a systematic analysis of the feasibility and reproducibility of these tools is lacking. Here we focused on InSITE GAL4 drivers that specifically label different subpopulations of olfactory neurons, particularly local interneurons (LNs), and genetically swapped the GAL4 domain for LexA, GAL80 or QF at the same locus. We found that the major utility-limiting factor for these genetic swaps is that many do not fully reproduce the original GAL4 expression patterns. Different donors exhibit distinct efficacies for reproducing original GAL4 expression patterns. The successfully swapped lines reported here will serve as valuable reagents and expand the genetic toolkits of Drosophila olfactory circuit research.

N-Propargylated isatin-Mannich mono- and bis-adducts: synthesis and preliminary analysis of in vitro activity against Tritrichomonas foetus.

Cu(I)Cl promoted synthesis of N-propargylated-isatin Mannich mono- and bis-adducts with an extension towards the synthesis of N-propargylated-isatin-7-chloroquinoline conjugates was described. The synthesized scaffolds were evaluated for their in vitro activity against the veterinary protozoal pathogen Tritrichomonas foetus and cytotoxicity against human prostate (PC-3) cancer cell line. The preliminary evaluation data revealed the enhancement in the activity profiles with the introduction of 7-chloroquinoline ring with the most active conjugates 7a, 7c and 7d exhibiting an IC50 of 22.2, 11.3 and 24.5 µM respectively against T. foetus and minimal toxicity against human prostate (PC-3) cell lines.

Use of bisphosphonates and the risk of osteonecrosis among cancer patients: a systemic review and meta-analysis of the observational studies.

PURPOSE: This study aims to systematically review observational studies evaluating the use of bisphosphonates (BPs) and risk of osteonecrosis of the jaw (ONJ) among cancer patients. METHODS: PubMed, Embase, and Cochrane Library were screened from database inception to Aug 2012. Two reviewers independently identified cohort and case-control studies evaluating the use of oral or intravenous (IV) BPs and the risk of ONJ and extracted the characteristics of the studies and risk estimates. Pooled estimates of odds ratios and 95 % confidence intervals were derived by random effects meta-analysis. Subgroup analyses were carried out according to patients' characteristics and route of BP use. RESULTS: We identified eight studies, including 1,389 cases and 569,620 controls. Use of BPs was associated with a significantly increased risk of ONJ (odds ratio (OR) 4.25; 95 % confidence interval (CI) 3.67-5.36; I (2) = 0 %). The summary OR was 4.22 (95 % CI 3.21-5.54; I (2) = 0 %) for adjusted studies. IV BPs were associated with higher risk (OR 4.27; 95 % CI 3.38-5.40; I (2) = 0 %) than oral BPs (OR 1.18; 95 % CI 0.89-1.56; I (2) = 0 %). Hospital-based studies were associated with higher risk estimates than population-based studies. CONCLUSION: The available evidence suggests that use of BPs in cancer patients is associated with a substantial risk for ONJ. Patients receiving IV BP are at the highest risk. It is important to assess oral health before initiating therapy and to avoid dental procedures during the active phase of intravenous BP therapy.

Effectiveness of external respiratory surrogates for in vivo liver motion estimation.

PURPOSE: Due to low frame rate of MRI and high radiation damage from fluoroscopy and CT, liver motion estimation using external respiratory surrogate signals seems to be a better approach to track liver motion in real-time for liver tumor treatments in radiotherapy and thermotherapy. This work proposes a liver motion estimation method based on external respiratory surrogate signals. Animal experiments are also conducted to investigate related issues, such as the sensor arrangement, multisensor fusion, and the effective time period. METHODS: Liver motion and abdominal motion are both induced by respiration and are proved to be highly correlated. Contrary to the difficult direct measurement of the liver motion, the abdominal motion can be easily accessed. Based on this idea, our study is split into the model-fitting stage and the motion estimation stage. In the first stage, the correlation between the surrogates and the liver motion is studied and established via linear regression method. In the second stage, the liver motion is estimated by the surrogate signals with the correlation model. Animal experiments on cases of single surrogate signal, multisurrogate signals, and long-term surrogate signals are conducted and discussed to verify the practical use of this approach. RESULTS: The results show that the best single sensor location is at the middle of the upper abdomen, while multisurrogate models are generally better than the single ones. The estimation error is reduced from 0.6 mm for the single surrogate models to 0.4 mm for the multisurrogate models. The long-term validity of the estimation models is quite satisfactory within the period of 10 min with the estimation error less than 1.4 mm. CONCLUSIONS: External respiratory surrogate signals from the abdomen motion produces good performance for liver motion estimation in real-time. Multisurrogate signals enhance estimation accuracy, and the estimation model can maintain its accuracy for at least 10 min. This approach can be used in practical applications such as the liver tumor treatment in radiotherapy and thermotherapy.