Efficacy of Radiofrequency Thermocoagulation of the Thoracic Sympathetic Nerve versus Chemical Excision in Pain Caused by Raynaud's Disease.

Objective: To investigate the effectiveness and safety of computed tomography (CT)-guided radiofrequency thermocoagulation (RFTC) of the thoracic sympathetic nerve versus chemical resection (CTS) for the treatment of pain caused by Raynaud's disease. Methods: Patients who underwent CTS or thoracic sympathetic nerve RFTC between March 2012 and March 2021 were enrolled in this retrospective study. There were 28 cases in the alcohol group (Group A) and 44 in the radiofrequency group (Group R). Visual analog scores (VAS) were collected from patients at different time points, as well as preoperative and postoperative finger end perfusion index (PI) and hand temperature (T). The efficiency, postoperative recurrence rate, complications, and improvement in postoperative quality of life were observed in both groups. Results: Pain scores at different follow-up times after surgery decreased in both groups compared to the preoperative period (P < 0.05). Postoperative T and PI were higher in both groups than preoperatively all (P < 0.05). The recurrence rate was higher in the R group than in the A group. Postoperative complications were observed in 13.6% and 25% of patients in groups R and A, respectively. Meanwhile, the postoperative quality of life improved in both groups, but the radiofrequency (RF) group was better than the alcohol group in terms of improvement in quality of life (P < 0.05). Conclusion: Both CT-guided CTS and RFTC of the thoracic sympathetic nerve provided good treatment outcomes. However, the RF group was superior to the alcohol group in terms of complication rate and quality of life improvement.

Physiological analysis of the improved ε-polylysine production induced by reactive oxygen species.

INTRODUCTION: Epsilon-poly-L-lysine (ε-PL) is produced by Streptomyces species in acidic and aerobic conditions, which inevitably induces rapid generation of reactive oxygen species (ROS). The devastating effects of ROS on biomolecules and cell vitality have been well-studied, while the positive effects of ROS are rarely reported. RESULTS: In this study, we found that a proper dose of intracellular ROS (about 3.3 µmol H2O2 /g DCW) could induce a physiological modification to promote the ε-PL production (from 1.2 to 1.5 g/L). It resulted in larger sizes of colony and mycelial pellets as well as vibrant, aggregated, and more robust mycelia, which were of high capability of ROS detoxication. Physiological studies showed that appropriate doses of ROS activated the metabolism of the pentose phosphate pathway at both transcriptional and enzymatic levels, which was beneficial for biomass accumulation. The biosynthesis of lysine was also promoted in terms of transcriptional regulatory overexpression, increased transcription and enzymatic activity of key genes, larger pools of metabolites in the TCA cycle, replenishment pathway, and diaminoheptanedioic acid pathway. In addition, energy provision was ensured by activated metabolism of the TCA cycle, a larger pool of NADH, and higher activity of the electron transport system. Increased transcription of HrdD and pls further accelerated the ε-PL biosynthesis. SIGNIFICANCE: These results indicated that ROS at proper intracellular dose could act as an inducing signal to activate the ε-PL biosynthesis, which laid a foundation for further process regulation to maintain optimal ROS dose in industrial ε-PL production and was of theoretical and practical significance. KEY POINTS: • A proper dose of intracellular ROS positively influences the ε-PL production. • Proper dose of ROS enhanced the mycelial activity and antioxidative capability. • ROS increased lysine synthesis metabolism, energy provision and pls expression.

Transcriptome and Metabolome Analysis Revealing the Improved ε-Poly-l-Lysine Production Induced by a Microbial Call from Botrytis cinerea.

ε-Poly-l-lysine (ε-PL) is a wide-spectrum antimicrobial agent, while its biosynthesis-inducing signals are rarely reported. This study found that Botrytis cinerea extracts could act as a microbial call to induce a physiological modification of Streptomyces albulus for ε-PL efficient biosynthesis and thereby resulted in ε-PL production (34.2 g/liter) 1.34-fold higher than control. The elicitors could be primary isolated by ethanol and butanol extraction, which resulted in more vibrant, aggregate and stronger mycelia. The elicitor-derived physiological changes focused on three aspects: ε-PL synthase, energy metabolism, and lysine biosynthesis. After elicitor addition, upregulated sigma factor hrdD and improved transcription and expression of pls directly contributed to the high ε-PL productivity; upregulated genes in tricarboxylic acid (TCA) cycle and energy metabolism promoted activities of citrate synthase and the electron transport system; in addition, pool enlargements of ATP, ADP, and NADH guaranteed the ATP provision for ε-PL assembly. Lysine biosynthesis was also increased based on enhancements of gene transcription, key enzyme activities, and intracellular metabolite pools related to carbon source utilization, the Embden-Meyerhof pathway (EMP), the diaminopimelic acid pathway (DAP), and the replenishment pathway. Interestingly, the elicitors stimulated the gene transcription for the quorum-sensing system and resulted in upregulation of genes for other antibiotic production. These results indicated that the Botrytis cinerea could produce inducing signals to change the Streptomyces mycelial physiology and accelerate the ε-PL biosynthesis. IMPORTANCE This work identified the role of microbial elicitors on ε-PL production and disclosed the underlying mechanism through analysis of gene transcription, key enzyme activities, and intracellular metabolite pools, including transcriptome and metabolome analysis. It was the first report for the inducing effects of the "microbial call" to Streptomyces albulus and ε-PL biosynthesis, and these elicitors could be potentially obtained from decayed fruits infected by Botrytis cinerea; hence, this may be a way of turning a biohazard into bioproduct wealth. This study provided a reference for application of microbial signals in secondary metabolite production, which is of theoretical and practical significance in industrial antibiotic production.

Thuricins: Novel Leaderless Bacteriocins with Potent Antimicrobial Activity Against Gram-Positive Foodborne Pathogens.

Bacteriocins are bacterial-derived peptides that exhibit antimicrobial activity and can be used as food preservatives. Here, using the indicator strain Bacillus cereus CMCC63301, we screened and identified a Bacillus thuringiensis LX43 strain that exhibits potent antimicrobial activity and harbors a putative leaderless bacteriocin gene cluster (thn gene cluster). Five novel leaderless bacteriocins, thuricin A1, A2, A3, A4, and A5, encoded by the thn gene cluster, were purified and identified. Thuricin A5 was regarded as a representative and showed remarkable antimicrobial activity against foodborne pathogens B. cereus, Clostridium perfringens, Listeria monocytogenes, and Staphylococcus aureus, likely by damaging their cell envelope. Moreover, thuricin A5 displayed good thermal and pH stability, with no hemolytic activity and cytotoxicity, indicating its wide applicability and biosafety. Furthermore, thuricin A5 effectively inhibited or eradicated foodborne pathogens in skim milk at 25 °C in a dose-dependent manner, affirming its potential for use as a novel biopreservative in foods.

Four Novel Leaderless Bacteriocins, Bacin A1, A2, A3, and A4 Exhibit Potent Antimicrobial and Antibiofilm Activities against Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major bacterial pathogen that causes hospital- and community-acquired infections. Owing to its multidrug resistance, it is imperative to develop new antimicrobial agents to treat MRSA infections. In this study, using genome mining analysis and a culture-based screening method to detect bacteriocin activity, we screened a strain, Bacillus sp. TL12, which harbored a putative leaderless bacteriocin gene cluster (bac gene cluster) and exhibited potent anti-MRSA activity. The antimicrobial agents, products of the bac gene cluster, were purified and identified as four novel leaderless bacteriocins: bacin A1, A2, A3, and A4. Bacin A2 was evaluated as a representative antimicrobial agent and showed remarkable antimicrobial activity against S. aureus, MRSA, and the foodborne pathogens Listeria monocytogenes and Bacillus cereus. Mechanistic experiments revealed that bacin A2 damaged cell membranes and exhibited bactericidal activity against MRSA. Bacin A2 effectively inhibited the formation of S. aureus and MRSA biofilms (>0.5× MIC) and killed the cells in their established biofilms (>4× MIC). The hemolytic and NIH/3T3 cytotoxicity assay results for bacin A2 confirmed its biosafety. Thus, bacins have potential as alternative antimicrobial agents for treating MRSA infections. IMPORTANCE Methicillin-resistant Staphylococcus aureus (MRSA) is a major human pathogen that is difficult to treat because of its resistance to several widely used antibiotics. The present study aimed to identify novel anti-MRSA bacteriocins in a prominent producer of bacteriocins, Bacillus cereus group. Four novel leaderless bacteriocins, bacin A1, A2, A3, and A4, which show potent bactericidal effect against S. aureus and MRSA, were identified in Bacillus sp. TL12. Moreover, bacins inhibited biofilm formation and killed cells in the established biofilms of S. aureus and MRSA. These findings suggest that bacins are promising alternatives to treat MRSA infections.

Long-Term Follow-Up Safety and Effectiveness of CT-Guided Radiofrequency Thermocoagulation of Sphenopalatine Ganglion in Refractory Headache Treatment.

OBJECTIVE: The purpose of this study was to evaluate the effectiveness and safety of computed tomography (CT)-guided radiofrequency thermocoagulation (RFTA) of the sphenopalatine ganglion (SPG) for patients with refractory headache. METHODS: A total of 14 patients with refractory migraine and 10 patients with cluster headache (CH) who underwent CT-guided SPG RF between May 2019 and August 2021 at the Jiaxing First Hospital, located in Jiaxing City, Zhejiang Province, China, were included and analyzed in this retrospective cohort study. Pain score, sleep quality scores, and treatment effects were observed before operation as well as 1 day and 1, 3, 6, 12, and 24 months after surgery. Also, the incidence of facial numbness at different timepoints after operation was evaluated. RESULTS: The frequency and duration of attacks decreased after treatment in patients with migraine, and the shortening of the cluster period and the prolongation of the remission period after treatment in patients with CH indicated that the treatment was effective. The numeric rating scale (NRS) ranged from 0 to 10, where 0 meant no pain and 10 meant the worst imaginable pain. The NRS of patients at 1 day and 1, 3, 6, 12, and 24 months after surgery was significantly lower than before operation (P < 0.05). The treatment of patients with migraine and CH was effective. The overall incidence of numbness in patients with migraine and the total incidence of numbness in patients with CH was recorded. The total incidence of numbness decreased gradually, but no significant difference was detected in the incidence of numbness between the two groups (P > 0.05). No serious adverse reactions, such as orthostatic hypertension, intracranial infection, and visual disturbance, occurred in the patients after operation. CONCLUSION: CT-guided RFTA of the SPG significantly relieves headache symptoms in patients with refractory migraine and CH. It has the advantages of rapid onset, long duration, and a safe and reliable treatment process, making it worthy of clinical application.

Trigeminal extracranial thermocoagulation along with patient-controlled analgesia with esketamine for refractory postherpetic neuralgia after herpes zoster ophthalmicus: A case report.

BACKGROUND: Primary trigeminal neuralgia can achieve satisfactory results through clinical treatment and intervention. The pathogenesis of neuralgia caused by varicella-zoster virus infection of the trigeminal nerve is more complex, and it is still difficult to relieve the pain in some patients simply by drug treatment or surgical intervention. CASE SUMMARY: A 66-year-old woman was hospitalized with herpetic neuralgia after herpes zoster ophthalmicus (varicella-zoster virus infects the ophthalmic branch of the trigeminal nerve). On admission, the patient showed spontaneous, electric shock-like and acupuncture-like severe pain in the left frontal parietal region, and pain could be induced by touching the herpes area. The numerical rating scale (NRS) was 9. There was no significant pain relief after pulsed radiofrequency and thermocoagulation of the ophthalmic branch of the trigeminal nerve. Combined with patient-controlled intravenous analgesia (PCIA) with esketamine, neuralgia was significantly improved. The patient had no spontaneous pain or allodynia at discharge, and the NRS score decreased to 2 points. The results of follow-up 2 mo after discharge showed that the NRS score was ≤ 3, and the Pittsburgh Sleep Quality Index score was 5 points. There were no adverse reactions. CONCLUSION: Trigeminal extracranial thermocoagulation combined with esketamine PCIA may be a feasible method for the treatment of refractory herpetic neuralgia after herpes zoster ophthalmicus.

Toyoncin, a Novel Leaderless Bacteriocin That Is Produced by Bacillus toyonensis XIN-YC13 and Specifically Targets B. cereus and Listeria monocytogenes.

Bacteriocins have attracted increasing interest because of their potential as natural preservatives. Recent studies showed that the Bacillus cereus group is a prominent producer of bacteriocins. Using a laboratory-based screening strategy, we identified a strain in the B. cereus group, Bacillus toyonensis XIN-YC13, with antimicrobial activity against B. cereus. A novel, 70-amino-acid-long leaderless bacteriocin, toyoncin, was purified from the culture supernatant of strain XIN-YC13, and its molecular mass was found to be 7,817.1012 Da. Toyoncin shares no similarity with any other known bacteriocins, and its N-terminal amino acid is formylmethionine rather than methionine. Toyoncin shows good pH and heat stability and exhibits specific antimicrobial activity against two important foodborne pathogens, B. cereus and Listeria monocytogenes. Additionally, toyoncin exerts bactericidal activity and induces cell membrane damage. Toyoncin can also inhibit the outgrowth of B. cereus spores. Preservation assays showed that toyoncin effectively suppressed or eradicated B. cereus and L. monocytogenes in pasteurized skim milk. These results suggest that toyoncin can be used as a new biopreservative against B. cereus and L. monocytogenes in the food industry. IMPORTANCE We identified a novel leaderless bacteriocin, toyoncin, produced by B. toyonensis XIN-YC13. Toyoncin shows good pH and heat stability, and it has specific antimicrobial activity against B. cereus and L. monocytogenes (two important foodborne pathogens), likely by destroying their cell membrane integrity. Toyoncin inhibited the outgrowth of B. cereus spores and effectively inhibited or eliminated B. cereus and L. monocytogenes in a milk model system. These results indicate the potential of toyoncin as a food preservative.

In Silico Analysis Highlights the Diversity and Novelty of Circular Bacteriocins in Sequenced Microbial Genomes.

Consumer demand for "fresh food" with no chemical preservatives has prompted researchers to pay more attention to natural antimicrobial peptides such as bacteriocins. Nisin is currently the most widely used food biopreservative among the bacteriocins; however, its applications are restricted due to its low stability at neutral and alkaline pH values. Circular bacteriocins have potent antimicrobial activity against foodborne pathogens, show exceptional stability, and have great potential to be developed as biopreservatives. Here, we take advantage of the precursor peptides of 15 reported circular bacteriocins to devise an in silico approach to identify potential circular bacteriocins in sequenced microbial genomes. A total of nearly 7,000 putative precursor peptides were identified from 86 species of bacteria and further classified into 28 groups based on their amino acid similarity. Among the groups, 19 showed low similarity (less than 50%) to any known precursor peptide of circular bacteriocins. One novel circular bacteriocin in group 11, cerecyclin, showed the highest identity (34%) to the known circular bacteriocin enterocin NKR-5-3B and was selected for verification. Cerecyclin showed antimicrobial activity against several Gram-positive bacteria, inhibited the outgrowth of Bacillus cereus spores, and did not exhibit hemolysis activity. Moreover, it showed 4-fold- to 8-fold-higher antimicrobial activity against B. cereus and Listeria monocytogenes than nisin A. Cerecyclin also had increased stability compared to nisin A under neutral or alkaline conditions. This work not only identified a promising food biopreservative but also provided a rich source for novel circular bacteriocins.IMPORTANCE Circular bacteriocins are promising biopreservatives, and it is important to identify more novel circular bacteriocins to enhance the current arsenal of antimicrobials. In this study, we used an in silico approach to identify a large number of novel circular bacteriocins and classified these bacteriocins into 28 groups rather than the 2 groups that were described in previous studies. Nineteen groups were novel and had low similarity (less than 50%) to any known precursor peptides of circular bacteriocins; this finding greatly expands the awareness of the novelty and diversity of circular bacteriocins. A novel circular bacteriocin which we named cerecyclin was identified in the B. cereus group; this circular bacteriocin had great antimicrobial activity against some foodborne pathogens and showed extreme stability. This study not only identified a promising food biopreservative but also provided a rich source for the identification of novel circular bacteriocins and the development of new biopreservatives.

Thusin, a Novel Two-Component Lantibiotic with Potent Antimicrobial Activity against Several Gram-Positive Pathogens.

Due to the rapidly increasing prevalence of multidrug-resistant bacterial strains, the need for new antimicrobial drugs to treat infections has become urgent. Bacteriocins, which are antimicrobial peptides of bacterial origin, are considered potential alternatives to conventional antibiotics and have attracted widespread attention in recent years. Among these bacteriocins, lantibiotics, especially two-component lantibiotics, exhibit potent antimicrobial activity against some clinically relevant Gram-positive pathogens and have potential applications in the pharmaceutical industry. In this study, we characterized a novel two-component lantibiotic termed thusin that consists of Thsα, Thsß, and Thsß' (mutation of Thsß, A14G) and that was isolated from a B. thuringiensis strain BGSC 4BT1. Thsα and Thsß (or Thsß') exhibit optimal antimicrobial activity at a 1:1 ratio and act sequentially to affect target cells, and they are all highly thermostable (100°C for 30 min) and pH tolerant (pH 2.0 to 9.0). Thusin shows remarkable efficacy against all tested Gram-positive bacteria and greater activities than two known lantibiotics thuricin 4A-4 and ticin A4, and one antibiotic vancomycin against various bacterial pathogens (Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus (MRSA), Staphylococcus sciuri, Enterococcus faecalis, and Streptococcus pneumoniae). Moreover, thusin is also able to inhibit the outgrowth of B. cereus spores. The potent antimicrobial activity of thusin against some Gram-positive pathogens indicates that it has potential for the development of new drugs.

Three Novel Lantibiotics, Ticins A1, A3, and A4, Have Extremely Stable Properties and Are Promising Food Biopreservatives.

Lantibiotics are antimicrobial peptides with potential applications as the next generation of antimicrobials in the food industry and/or the pharmaceutical industry. Nisin has successfully been used as a food preservative for over 40 years, but its major drawback is its limited stability under neutral and alkaline pH conditions. To identify alternatives with better biochemical properties, we screened more than 100 strains of the Bacillus cereus group. Three novel lantibiotics, ticins A1 (4,062.98 Da), A3 (4,048.96 Da), and A4 (4,063.02 Da), which were highly thermostable (121°C for 30 min) and extremely pH tolerant (pH 2.0 to 9.0), were identified in Bacillus thuringiensis BMB3201. They all showed potent antimicrobial activities against all tested Gram-positive bacteria and greater activities than those of nisin A against Bacillus cereus and Listeria monocytogenes, two important foodborne pathogens. These three novel lantibiotics, with their extremely stable properties and potent antimicrobial activities, have the potential for use as biopreservatives.

The Bacillus cereus group is an excellent reservoir of novel lanthipeptides.

Lantibiotics are ribosomally synthesized peptides that contain multiple posttranslational modifications. Research on lantibiotics has increased recently, mainly due to their broad-spectrum antimicrobial activity, especially against some clinical Gram-positive pathogens. Many reports about various bacteriocins in the Bacillus cereus group have been published, but few were about lantibiotics. In this study, we identified 101 putative lanthipeptide gene clusters from 77 out of 223 strains of this group, and these gene clusters were further classified into 20 types according to their gene organization and the homologies of their functional genes. Among them, 18 types were novel and have not yet been experimentally verified. Two novel lantibiotics (thuricin 4A-4 and its derivative, thuricin 4A-4D) were identified in the type I-1 lanthipeptide gene cluster and showed activity against all tested Gram-positive bacteria. The mode of action of thuricin 4A-4 was studied, and we found that it acted as a bactericidal compound. The transcriptional analysis of four structural genes (thiA1, thiA2, thiA3, and thiA4) in the thuricin 4A gene cluster showed that only one structural gene, thiA4, showed efficient transcription in the exponential growth phase; the other three structural genes did not. In addition, the putative transmembrane protein ThiI was responsible for thuricin 4A-4 immunity. Genome analysis and functional verification illustrated that B. cereus group strains were a prolific source of novel lantibiotics.