CDK5-USP30 signaling pathway regulates MAVS-mediated inflammation via suppressing mitophagy in MPTP/MPP+ PD model.

The discovery of MPTP, an industrial chemical and contaminant of illicit narcotics, which causes parkinsonism in humans, non-human primates and rodents, has led to environmental pollutants exposure being convicted as key candidate in Parkinson's disease (PD) pathogenesis. Though MPTP-induced mitochondrial dysfunction and neuroinflammation are mainly responsible for the causative issue of MPTP neurotoxicity, the underlying mechanism involved remains unclear. Here, we reveal a novel signaling mechanism of CDK5-USP30-MAVS regulating MPTP/MPP+ induced PD. MPP+ (the toxic metabolite of MPTP) treatment not only led to the increased protein levels of USP30 but also to mitophagy inhibition, mitochondrial dysfunction, and MAVS-mediated inflammation in BV2 microglial cells. Both mitophagy stimulation (Urolithin A administration) and USP30 knockdown relieved MAVS-mediated inflammation via restoring mitophagy and mitochondrial function in MPP+-induced cell model. Notably, MPTP/MPP+-induced CDK5 activation regulated USP30 phosphorylation at serine 216 to stabilize USP30. Moreover, CDK5-USP30 pathway promoted MAVS-mediated inflammation in MPTP/MPP+-induced PD model. Inhibition of CDK5 not only had a protective effect on MPP+-induced cell model of PD via suppressing the upregulation of USP30 and the activation of MAVS inflammation pathway in vitro, but also prevented neurodegeneration in vivo and alleviated movement impairment in MPTP mouse model of PD. Overall, our study reveal that CDK5 blocks mitophagy through phosphorylating USP30 and activates MAVS inflammation pathway in MPTP/MPP+-induced PD model, which suggests that CDK5-USP30-MAVS signaling pathway represents a valuable treatment strategy for PD induced by environmental neurotoxic pollutants in relation to MPTP.

Comparative analysis of antioxidant activities and chemical compositions in the extracts of different edible parts from Camellia tetracocca Zhang (C. tetracocca) with two distinct color characteristics.

The Camellia tetracocca Zhang is a rare and ancient plant, exclusively found in the vicinity of Puan County, Guizhou Province, China. According to leaf color, two distinct variations have been identified: purple C. tetracocca Zhang (PCTZ) and green C. tetracocca (GCTZ). This research was conducted to investigate the antioxidant activities and chemical compositions of different edible parts of PCTZ and GCTZ. Antioxidant activity was evaluated using DPPH, ABTS, HSA, and T-AOC assays, while the content of compounds was determined by HPLC. The findings demonstrated that the antioxidant capacity of PCTZ leaves is significantly superior to that of GCTZ leaves. Notably, theacrine, a rare compound, contains up to 2.075% in PCTZ leaves, indicating potential as a novel natural antidepressant and antioxidant. In conclusion, PCTZ is a distinctive tea beverage and a valuable genetic material for tea tree breeding due to its high theacrine and low caffeine characteristics.

Clinical Report of a Rare Mucinous Staphylococcus Aureus Infection.

BACKGROUND: Staphylococcus aureus is the most common pathogen in suppurative infection, which can cause local suppurative infection, pneumonia, etc. A case of double renal calculi complicated with chronic renal insufficiency and mucinous Staphylococcus aureus infection was analyzed and discussed. METHODS: Bacterial culture, identification, and next-generation sequencing. RESULTS: The mucous colony was identified as Staphylococcus aureus, and the condition improved after symptomatic treatment. CONCLUSIONS: Mucinous Staphylococcus is a rare clinical microorganism, which needs to be verified by experiments to avoid false negative results. Genetic sequencing is used to identify strains if necessary.

Predicting Blood Glucose Concentration after Short-Acting Insulin Injection Using Discontinuous Injection Records.

Diabetes is an increasingly common disease that poses an immense challenge to public health. Hyperglycemia is also a common complication in clinical patients in the intensive care unit, increasing the rate of infection and mortality. The accurate and real-time prediction of blood glucose concentrations after each short-acting insulin injection has great clinical significance and is the basis of all intelligent blood glucose control systems. Most previous prediction methods require long-term continuous blood glucose records from specific patients to train the prediction models, resulting in these methods not being used in clinical practice. In this study, we construct 13 deep neural networks with different architectures to atomically predict blood glucose concentrations after arbitrary independent insulin injections without requiring continuous historical records of any patient. Using our proposed models, the best root mean square error of the prediction results reaches 15.82 mg/dL, and 99.5% of the predictions are clinically acceptable, which is more accurate than previously proposed blood glucose prediction methods. Through the re-validation of the models, we demonstrate the clinical practicability and universal accuracy of our proposed prediction method.

Isolation, characterization and comparison of lytic Epseptimavirus phages targeting Salmonella.

This study describes the characterization and genomic analysis of six lytic Salmonella phages. To examine the feasibility of using these phages as biocontrol agents, we analyzed their genomes and compared them to those of similar phages. These six phages belong to genus Epseptimavirus, family Demerecviridae. We identified the genes of these six phages by comparing their genomes with those of three type phages in subfamily Markadamsvirinae. All six phages examined in this study were obligately lytic and did not carry undesirable genes. Two phages (vB_SalS_1-23 and vB_SalS_3-29) were selected as the representative phages for general characterization and physiological tests. The biocontrol efficacy of the representative phages was determined by comparing the viable counts of recovered host Salmonella ser. Newlands ZC-S1 from treatment and phage-free control samples. The biocontrol experiment showed that the representative phages were able to reduce the counts of ZC-S1 to below 2 log10 CFU/mL (~4.3 log10 CFU/mL reduction) at 3 h post-infection at 37 °C. Furthermore, we investigated the application of these two phages in the control of ZC-S1 contamination in chicken products and on eggshells. When applied to the surfaces of the samples, the phage cocktail (MOI = 100) reduced the ZC-S1 count to below 2 log10 CFU/mL on chicken skin and to undetectable levels (1 log10 CFU/mL) in chicken breast meat, ground chicken meat and eggshell samples (p < 0.01). Compared to the initial experiment, the phage cocktail reduced the ZC-S1 count by 2-4.08 log10 CFU/mL when applied at an MOI = 1 (except in the ground chicken meat group) and by 4.48-5.67 log10 CFU/mL at an MOI = 100 after 7 h. In conclusion, these two phages with lytic effects show a high potential to inhibit the growth of Salmonella contaminants and can be used as candidate biocontrol agents.

The endolysin of the Acinetobacter baumannii phage vB_AbaP_D2 shows broad antibacterial activity.

The emergence and rapid spread of multidrug-resistant bacteria has induced intense research for novel therapeutic approaches. In this study, the Acinetobacter baumannii bacteriophage D2 (vB_AbaP_D2) was isolated, characterized and sequenced. The endolysin of bacteriophage D2, namely Abtn-4, contains an amphipathic helix and was found to have activity against multidrug-resistant Gram-negative strains. By more than 3 log units, A. baumannii were killed by Abtn-4 (5 µM) in 2 h. In absence of outer membrane permeabilizers, Abtn-4 exhibited broad antimicrobial activity against several Gram-positive and Gram-negative bacteria, such as Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Enterococcus and Salmonella. Furthermore, Abtn-4 had the ability to reduce biofilm formation. Interestingly, Abtn-4 showed antimicrobial activity against phage-resistant bacterial mutants. Based on these results, endolysin Abtn-4 may be a promising candidate therapeutic agent for multidrug-resistant bacterial infections.

Hydroxyapatite-incorporation improves bone formation on endosseous PEEK implant in canine tibia.

BACKGROUND: Poly Ether Ether Ketone (PEEK) has been considered as a potential alternative material for endosseous dental implants, for its low elastic modulus, biocompatibility, and low cost in customized device manufacture. Hydroxyapatite-incorporation is supposed to improve the poor osseointegration of PEEK. METHODS: In the present study we analyzed the in vivo response of hydroxyapatite-incorporated PEEK (PEEK-HA) implants in canine tibia. PEEK-HA and PEEK implants were implanted and were examined 4 weeks and 12 weeks after implantation with radiology and histology. Commercial titanium dental implants served as controls. RESULTS: The ratio of bone volume to tissue volume of PEEK-HA implants was higher than that of PEEK implants 4 weeks after implantation in the µ-CT analysis. The bone implant contact of PEEK and PEEK-HA implants showed no statistical difference in the histological examination, but newly-formed bone around PEEK-HA implants showed more signs of mineralization than that around PEEK implants. CONCLUSION: The study suggested that bone formation was improved with hydroxyapatite-incorporation in PEEK. Hydroxyapatite-incorporated PEEK implants may represent a potential material for endosseous dental implant.

A photo-sensitizable phage for multidrug-resistant Acinetobacter baumannii therapy and biofilm ablation.

Antibiotic abuse causes the emergence of bacterial resistance. Photodynamic antibacterial chemotherapy (PACT) has great potential to solve serious bacterial resistance, but it suffers from the inefficient generation of ROS and the lack of bacterial targeting ability. Herein, a unique cationic photosensitizer (NB) and bacteriophage (ABP)-based photodynamic antimicrobial agent (APNB) is developed for precise bacterial eradication and efficient biofilm ablation. Thanks to the structural modification of the NB photosensitizer with a sulfur atom, it displays excellent reactive oxygen species (ROS)-production ability. Moreover, specific binding to pathogenic microorganisms can be provided by bacteriophages. The developed APNB has multiple functions, including bacteria targeting, near-infrared fluorescence imaging and combination therapy (PACT and phage therapy). Both in vitro and in vivo experiments prove that APNB can efficiently treat A. baumannii infection. Particularly, the recovery from A. baumannii infection after APNB treatment is faster than that with ampicillin and polymyxin B in vivo. Furthermore, the strategy of combining bacteriophages and photosensitizers is employed to eradicate bacterial biofilms for the first time, and it shows the excellent biofilm ablation effect as expected. Thus, APNB has huge potential in fighting against multidrug-resistant bacteria and biofilm ablation in practice.

Efficacy of a phage cocktail in controlling phage resistance development in multidrug resistant Acinetobacter baumannii.

The control and treatment of multidrug resistant pathogens infections has become a grand challenge for clinicians worldwide. Virulent phage has long been considered as an effective bactericidal agent, which may be a potentially alternative to antibiotics. However, the rapid development of phage resistance seriously hinders the wide and continuous application of virulent phages. In this study, Acinetobacter baumannii phage vB_AbaS_D0 was isolated, characterized and used to control the phage resistance development in bacterial strains. Transmission electron microscopy analysis of vB_AbaS_D0 indicated it belonged to the Siphoviridae family with an icosahedral head. Its whole genome was 43, 051 bp in size, with a GC content of 45.48% and 55 putative open reading frames. The data showed that vB_AbaS_D0 was a virulent phage. Although vB_AbaS_D0 had a very weak bactericidal activity, a wide range of Acinetobacter baumannii strains were sensitive to it. The results suggested that the cocktail of vB_AbaS_D0 and another Acinetobacter baumannii phage vB_AbaP_D2 could improve the therapeutic efficacy in vivo and in vitro. The resistance mutation frequency of A. baumannii cells infected with D0 or phage cocktail was significantly lower than cells treated with D2 (P < 0.01). Phage therapy in the murine bacteremia model results showed that the percentage of phage resistant mutant occurrence in the phage D0 or cocktail treatment group was significantly lower than in phage D2 treatment group (P < 0.01).

Complete genome analysis of the novel Enterococcus faecalis phage vB_EfaS_AL3.

This work describes the characterization and genome annotation of a new lytic Enterococcus faecalis siphovirus, vB_EfaS_AL3 (referred to as AL3), isolated from wastewater samples collected in Liaoning Province, China. The genome of phage AL3 is composed of linear double-stranded DNA that is 40,789 bp in length with a G + C content of 34.84% and 61 putative protein-coding genes. Phylogenetic and comparative genomic analyses indicate that phage AL3 should be considered a novel phage.

Isolation and characterization of a bacteriophage and its potential to disrupt multi-drug resistant Pseudomonas aeruginosa biofilms.

A lytic Pseudomonas aeruginosa bacteriophage, vB_PaeM_LS1, was isolated and characterized herein. To examine the eligibility of bacteriophage vB_PaeM_LS1 as a therapeutic bacteriophage, we analysed its genome and compared it to similar bacteriophages. Genome of bacteriophage vB_PaeM_LS1 consisted of a linear, double-stranded DNA molecule 66,095 bp in length and with 55.7% G + C content. Neighbor-joining analysis of the large subunit terminase showed that bacteriophage vB_PaeM_LS1 had similarity to the Pbunavirus genus. The potential of the lytic bacteriophage to disrupt Pseudomonas aeruginosa biofilms was assessed by scanning electron microscopy and bacterial counts. This study revealed that the bacteriophage vB_PaeM_LS1 with its lytic effect showed a high potential impact on the inhibition of the growth of Pseudomonas aeruginosa biofilm formation.

Isolation and identification of Salmonella pullorum bacteriophage YSP2 and its use as a therapy for chicken diarrhea.

Salmonella pullorum is the major pathogen that is harmful to the poultry industry in developing countries, and the treatment of chicken diarrhea caused by S. pullorum has become increasingly difficult. In this study, a virulent bacteriophage YSP2, which was able to specifically infect Salmonella, was isolated and characterized. Phage YSP2 was classified in the Siphoviridae family and had a short latent period of 10 min. No bacterial virulence- or lysogenesis-related ORF is present in the YSP2 genome, making it eligible for use in phage therapy. Experiments in vivo investigated the potential use of phages as a therapy against diarrhea in chickens caused by S. pullorum in a chicken diarrhea model, demonstrating that a single oral administration of YSP2 (1 × 1010 PFU/mL, 80 µL/chicken) 2 h after S. pullorum oral administration at a double median lethal dose was sufficient to protect chickens against diarrhea. Gross inspection showed that YSP2 can effectively reduce organ damage and significantly relieve hemorrhage in the intestine and liver tissue. Moreover, YSP2 can maintain a high curative effect when diluted to 108 PFU/mL. In light of its therapeutic effect on chicken diarrhea, YSP2 may serve as an alternative treatment strategy for infections caused by S. pullorum.

A novel biomimetic composite substitute of PLLA/gelatin nanofiber membrane for dura repairing.

OBJECTIVE: Biomimetic design will significantly improve growth and regeneration of dural cells and tissue for better repairing effects and fewer complications in repairing the native dura. This study designed a novel composite, biomimetic substitute based on the characteristics of native dura extracellular matrix. METHODS AND RESULTS: This substitute is expected to rapidly induce cell adhesion, migration, and fast regeneration of neotissue. The material characteristics (contact angle, surface charge, and zeta potential were evaluated), in vitro biological characteristics (cell stretch, connections between cells, cell proliferation) and in vivo tissue regeneration capability of this substitute were evaluated, compared to those of collagen dura substitute, the mostly used dura substitute. The results showed that the surface properties of this composite substitute were more biomimetic to native extracellular matrix than collagen substitute did, together with better cytocompatibility, tissue ingrowth, and neoangiogenesis. This composite substitute further demonstrated in clinical case study its ideal repair effect with no CSF leakage or other adverse reactions. CONCLUSION: In conclusion, the new biomimetic composite substitute provides alternative substitute for dura repairing.

A biomimetic mesh for treating female stress urinary incontinence.

For patients with medium to severe incontinence, sub-urethral support surgery has a high cure rate, but using synthetic meshes leads to some complications such as mesh erosion/exposure and thigh pain. Autologous or acellular extracellular matrix grafts present few complications but have a high recurrence rate. Regensling™ is a new sling product made of a synthetic material with a biomimetic structure, aiming to provide long-term mechanical support with a lower complication rate. To assess the safety and effectiveness of Regensling™, both in vitro and in vivo experiments were performed. The mesh was implanted in the subcutaneous, intramuscular and sub-urethral regions of rabbits. At 4, 12, and 26 weeks post-implantation, the animals were executed and the implants were studied for their mechanical and biocompatible properties. Compared to the control material, the Regensling™ was covered by a thin layer of fibrous tissue with good compliance, and had a milder inflammatory response. During the period of the experiment, Regensling™ showed stable strength with an increasing trend over time.

Evaluation of efficacy and biocompatibility of a new absorbable synthetic substitute as a dural onlay graft in a large animal model.

BACKGROUND: Numerous dura substitutes are commercially available, but no absorbable synthetic dura repair product has been used for both onlay and suture applications. OBJECTIVE: The safety and effectiveness of a new absorbable synthetic substitute composed of Poly-L-lactide microfibers as onlay dural graft were evaluated. METHODS: Physical properties and performance of the microfibrous synthetic dural substitute implanted as an onlay or suturable grafts were compared with these commercial products, including CODMAN ETHISORB™ Dura Patch and DuraGen™ Dural Graft Matrix, in a canine duraplasty model. The cerebrospinal fluid (CSF) leakage, macroscopic and microscopic observation at 30 and 90 days after implantation were investigated. RESULTS: The absorbable synthetic dural substitute exhibited good wettability and conformability. When implanted as an onlay graft, it can prevent CSF leakage and integrate with the surrounding tissue to repair dural defects, indicating its good efficacy and biocompatibility as an onlay graft. CONCLUSION: Based on the excellent physical properties and performances mentioned above, the new absorbable synthetic substitute can be applied as a dural graft for both onlay and suturable applications.

Nanofibrous biomimetic mesh can be used for pelvic reconstructive surgery: A randomized study.

BACKGROUND: Implantation of nonabsorbable polypropylene (PP) mesh in the vagina is the main surgical treatment for pelvic organ prolapse (POP); however, clinical outcomes remain controversial and far from satisfactory. In particular, reducing the exposure or erosion of vaginal implants to obtain improved functional reconstruction is challenging. There is an urgent need for the development of new materials and/or products for POP treatment. A nanofibrous biomimetic mesh was recently developed to address this issue. OBJECTIVE: In this study, the basic properties of the newly developed mesh, including structural characteristics, mechanical properties, biological response of human umbilical cord mesenchymal stem cells in vitro, and tissue regeneration and biocompatibility in vivo, were evaluated and compared with those of Gynemesh™PS. METHODS: Scanning electron microscopy and uniaxial tensile methods were used to evaluate microstructure and mechanical properties, respectively. Mesenchymal stem cell growth on the meshes was observed by fluorescence microscopy to visualize the expression of enhanced red fluorescent protein. Twenty-four mature female Sprague Dawley rats were randomly assigned to two groups: group 1 (nanofibrous biomimetic mesh, Medprin, Germany, n=12) and group 2 (Gynemesh(TM)PS, Ethicon, USA; n=12). The posterior vaginal wall was incised from the introitus, and the mesh was then implanted. Three implants of each type were tested for 1, 4, 8 and 12 weeks. Connective tissue organization, inflammation, vascularization, and regenerated tissue were histologically assessed. RESULTS: The nanofibrous biomimetic mesh is a relatively heavy material and exhibited lower porosity than Gynemesh(TM)PS. The new mesh was stiffer than Gynemesh(TM)PS (p<0.001) but supported human umbilical cord mesenchymal stem cell attachment. Erosion of the grafts did not occur in any animal. The nanofibrous biomimetic mesh was encapsulated by a thicker layer of connective tissue and was associated with significantly greater inflammatory scores compared with Gynemesh(TM)PS. At 12 weeks, the vascularization of the new mesh was greater than that of Gynemesh(TM)PS (p<0.05). No significant difference in the thickness of the smooth muscle layer following implantation was observed between the two groups (p>0.05). CONCLUSIONS: The nanofibrous biomimetic mesh is a candidate for reinforcing pelvic reconstruction. The mesh could be improved by decreasing its weight and stiffness and increasing its porosity. This mesh could serve as a carrier for stem cells in future regenerative medicine and tissue engineering research.

A New Absorbable Synthetic Substitute With Biomimetic Design for Dural Tissue Repair.

Dural repair products are evolving from animal tissue-derived materials to synthetic materials as well as from inert to absorbable features; most of them lack functional and structural characteristics compared with the natural dura mater. In the present study, we evaluated the properties and tissue repair performance of a new dural repair product with biomimetic design. The biomimetic patch exhibits unique three-dimensional nonwoven microfiber structure with good mechanical strength and biocompatibility. The animal study showed that the biomimetic patch and commercially synthetic material group presented new subdural regeneration at 90 days, with low level inflammatory response and minimal to no adhesion formation detected at each stage. In the biological material group, no new subdural regeneration was observed and severe adhesion between the implant and the cortex occurred at each stage. In clinical case study, there was no cerebrospinal fluid leakage, and all the postoperation observations were normal. The biomimetic structure and proper rate of degradation of the new absorbable dura substitute can guide the meaningful reconstruction of the dura mater, which may provide a novel approach for dural defect repair.

Development of Dual Neurotrophins-Encapsulated Electrosupun Nanofibrous Scaffolds for Peripheral Nerve Regeneration.

Nerve growth factor (NGF) is widely used for repairing peripheral nerve injury because of its capability in dominating the survival, migration, proliferation, and differentiation of nerve cells. Monosialoganglioside (GM1), as another kind of nerve growth factor, works for regulating NGF function. In this study, GM1 and NGF were incorporated into the Poly(l-lactic acid-co-ε-caprolactone)/silk fibroin (PLCL/SF) nanofibers by the coaxial electrospinning. The fibers morphology and core­shell structure were characterized by SEM and TEM. The scaffolds demonstrated high tensile stress with good flexibility. In vitro cell viability studies indicated that the scaffolds incorporating both GM1 and NGF played a synergistic effect to enhance Schwann cells (SCs) proliferation and Pheochromocytoma (PC12) cells differentiation, in comparison to the scaffolds only incorporating NGF. Subsequently, the nanofibrous conduit scaffolds (NCSs) were evaluated in vivo in a rabbit sciatic nerve defect model. The NGF/GM1 incorporated NCSs group performed better nerve function recovery than single incorporated group, in consideration of the compound muscle action potential (CMAP) and nerve conduction velocity (NCV) results. Furthermore, hematoxylin and eosin (H&E) staining, toluidine blue (TB) staining, and transmission electron microscope (TEM) analysis displayed better nerve regeneration of NGF/GM1 incorporated NCSs both quantitatively and qualitatively. Therefore, the results indicated the dual neurotrophins-incorporated NCSs had potentials for the application in peripheral nerve repairing.

Physically facilitating drug-delivery systems.

Facilitated/modulated drug-delivery systems have emerged as a possible solution for delivery of drugs of interest to pre-allocated sites at predetermined doses for predefined periods of time. Over the past decade, the use of different physical methods and mechanisms to mediate drug release and delivery has grown significantly. This emerging area of research has important implications for development of new therapeutic drugs for efficient treatments. This review aims to introduce and describe different modalities of physically facilitating drug-delivery systems that are currently in use for cancer and other diseases therapy. In particular, delivery methods based on ultrasound, electrical, magnetic and photo modulations are highlighted. Current uses and areas of improvement for these different physically facilitating drug-delivery systems are discussed. Furthermore, the main advantages and drawbacks of these technologies reviewed are compared. The review ends with a speculative viewpoint of how research is expected to evolve in the upcoming years.

Administration of high-dose macrophage colony-stimulating factor increases bone turnover and trabecular volume fraction.

Macrophage colony-stimulating factor (M-CSF) is a hematopoietic growth factor that plays a critical role in early osteoclastogenesis. To characterize the skeletal effects of M-CSF, we administered soluble M-CSF to mice. It was hypothesized that M-CSF would stimulate bone formation through coupled activity of osteoclasts and osteoblasts. Twenty-four male C57BL/6 J mice (n = 12/group, aged 7 weeks) received subcutaneous injections of human M-CSF [5 mg/(kg day)] or inert vehicle (VEH) for 21 days. M-CSF increased serum bone turnover markers (+57% TRAP-5b and +44% osteocalcin). Microcomputed tomography revealed an anabolic effect on tibial trabecular bone, with higher bone volume fraction (+35%), connectivity density (+79%), and number (+18%), as well as lower trabecular separation (-18%). M-CSF had no significant effect on cortical bone mineral content, geometry, or strength. There was no change in quantitative histomorphometry parameters of femoral cortical bone. These results reveal the complex, site-specific effects of M-CSF. In particular, we have demonstrated an anabolic effect of M-CSF on trabecular bone achieved through coupled activation of osteoblasts. However, in contrast to previous studies, M-CSF was found to have no effect on cortical bone. M-CSF was demonstrated to significantly influence both bone modeling and remodeling in relatively young animals.