A Novel White Laccase from Pleurotus ostreatus

Abstract Two laccase isoenzymes (POXA1 and POXA2) produced by Pleurotus ostreatus were purified and fully characterized. POXA1 and POXA2 are monomeric glycoproteins with 3 and 9% carbohydrate content, molecular masses of about 61 and 67 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, of about 54 and 59 kDa by gel filtration in native conditions, and of 61 kDa by matrix-assisted laser desorption ionization mass spectrometry (only for POXA1) and pI values of 6.7 and 4.0, respectively. The N terminus and three tryptic peptides of POXA1 have been sequenced, revealing clear homology with laccases from other microorganisms, whereas POXA2 showed a blocked N terminus. The stability of POXA2 as a function of temperature was particularly low, whereas POXA1 showed remarkable high stability with respect to both pH and temperature. Both enzymes oxidize syringaldazine and ABTS (2, 2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) together with a variety of different substituted phenols and aromatic amines with the concomitant reduction of oxygen, but POXA1 is unable to oxidize guaiacol. Both enzymes were strongly inhibited by sodium azide and thioglycolic acid but not by EDTA. UV/visible absorption spectra, atomic adsorption, and polarographic data indicated the presence of 4 copper atoms/mol of POXA2 but only one copper, two zinc, and one iron atoms were found/mol of POXA1. The neutral pI and the anomalous metal content of POXA1 laccase render this enzyme unique in its structural characteristics. The lack of typical absorbance at 600 nm allows its classification as a "white" laccase

A Study on the Antimicrobial and Antibiofilm Peptide 1018-K6 as Potential Alternative to Antibiotics against Food-Pathogen Salmonella enterica

Antimicrobial resistance has become one of the major global public health concerns, and it is indispensable to search for alternatives to conventional antibiotics. Recently, antimicrobial peptides have received great attention because of their broad-spectrum antimicrobial activity at relatively low concentrations, even against pathogens such as Salmonella enterica, which is responsible for most food-borne illnesses. This work aimed at evaluating the antimicrobial and antibiofilm activity of the innate defense peptide, named 1018-K6, against S. enterica. A total of 42 strains, belonging to three different subspecies and 32 serotypes, were included in this study. The antibiotic resistance profile of all the strains and the cytotoxic effects of 1018-K6 on mammalian fibroblast cells were also investigated. Results revealed that MIC (minimum inhibitory concentrations) and MBC (minimum bactericidal concentrations) values were in the ranges of 8–64 μg/mL and 16–128 μg/mL, respectively, although most strains (97%) showed MICs between 16 and 32 μg/mL. Moreover, sub-inhibitory concentrations of 1018-K6 strongly reduced the biofilm formation in several S. enterica strains, whatever the initial inoculum size. Our results demonstrated that 1018-K6 is able to control and manage S. enterica growth with a large potential for applications in the fields of active packaging and water disinfectantsThis research was funded by: Ministero dello Sviluppo Economico “Packaging Bioattivi e bIOsanitizzanti: Sviluppo di stratEgie iNnovaTIve ed ecososteNibili pEr L’Industria Alimentare—(BIO-SENTINEL) project, grant number F/200092/01-03/X45. Fondo per la Crescita Sostenibile—Sportello “Agrifood” PON I&C 2014-2020; Regione Campania-“Sviluppo di una tecnologia Intelligente contro spoilage ed agenti patogeni: dal peptide antimicrobico ad un PACKaging innovativo nella filiera ittica del Mediterraneo (iPACK-Med)project, grant number PO FEAMP 2014/2020, MISURA 1.26 “INNOVAZIONE” DD n. 266/2019-n. B68D19001380009; Ministero della Salute-“Attività battericida ed anti-biofilm di nano-sistemi ibridi coniugati con peptidi antimicrobici: una nuova strategia per la formulazione di bio-sanitizzanti contro ceppi patogeni resistenti”- Ricerca Corrente 2018 project, grant number IZS ME 06/18 RCS

Acylpeptide Hydrolase Inhibition as Targeted Strategy to Induce Proteasomal Down-Regulation

Acylpeptide hydrolase (APEH), one of the four members of the prolyl oligopeptidase class, catalyses the removal of N-acylated amino acids from acetylated peptides and it has been postulated to play a key role in protein degradation machinery. Disruption of protein turnover has been established as an effective strategy to down-regulate the ubiquitin-proteasome system (UPS) and as a promising approach in anticancer therapy

Genetic Pattern, Orthodontic and Surgical Management of Multiple Supplementary Impacted Teeth in a Rare, Cleidocranial Dysplasia Patient: A Case Report

Background: Cleidocranial dysplasia (CCD) is a rare, autosomal dominant skeletal dysplasia with a prevalence of one per million births. The main causes of CCD are mutations in the core-binding factor alpha-1 (CBFA1) or runt-related transcription factor-2 (RUNX2), located at the 6p21 chromosomal region. RUNX2 plays important roles in osteoblast differentiation, chondrocyte proliferation and differentiation, and tooth formation. The disease is characterized by clavicular aplasia or hypoplasia, Wormian bones, delayed closure of cranial suture, brachycephalic head, maxillary deficiency, retention of primary teeth, inclusion of permanent teeth, and multiple supernumerary teeth. Materials and Methods: A 22-year-old girl suffering from cleidocranial dysplasia with short stature, narrow shoulders, craniofacial manifestations (short face, broad forehead, etc.) and dental anomalies (different lower dental elements under eruption, supernumerary and impacted multiple teeth, etc.) was examined at our service (Complex Operative Unit of Odontostomatology of Policlinico of Bari). RX Orthopantomography (OPG) and cone beam computed tomography (CBCT) were requested to better assess the position of the supernumerary teeth and their relationships with others and to evaluate the bone tissue. Results: Under eruption was probably caused by dental interferences with supernumerary teeth; hence, extractions of supernumerary upper canines and lower premolars were performed under general anaesthesia. Surgery outcome was excellent with good tissue healing and improvements in the therapeutic possibilities with future orthodontics. Conclusions: The objective of this article is to give an update about radiological, clinical, and molecular features of CCD and to alert the health team about the importance of establishing an early diagnosis and an appropriate treatment in these patients to prevent impacted teeth complications and to offer them a better quality of life

Identification and Characterisation of a Novel Acylpeptide Hydrolase from Sulfolobus Solfataricus: Structural and Functional Insights

A novel acylpeptide hydrolase, named APEH-3Ss, was isolated from the hypertermophilic archaeon Sulfolobus solfataricus. APEH is a member of the prolyl oligopeptidase family which catalyzes the removal of acetylated amino acid residues from the N terminus of oligopeptides. The purified enzyme shows a homotrimeric structure, unique among the associate partners of the APEH cluster and, in contrast to the archaeal APEHs which show both exo/endo peptidase activities, it appears to be a “true” aminopeptidase as exemplified by its mammalian counterparts, with which it shares a similar substrate specificity. Furthermore, a comparative study on the regulation of apeh gene expression, revealed a significant but divergent alteration in the expression pattern of apeh-3Ss and apehSs (the gene encoding the previously identified APEHSs from S. solfataricus), which is induced in response to various stressful growth conditions. Hence, both APEH enzymes can be defined as stress-regulated proteins which play a complementary role in enabling the survival of S. solfataricus cells under different conditions. These results provide new structural and functional insights into S. solfataricus APEH, offering a possible explanation for the multiplicity of this enzyme in Archaea

Elastodontic Therapy of Hyperdivergent Class II Patients Using AMCOP® Devices: A Retrospective Study

Background: The management of a hyperdivergent growth pattern is one of the most challenging in orthodontics and different treatments are advocated. The present study analyses the effectiveness of elastodontic therapy with AMCOP® devices in treating children with hyperdivergent class II malocclusion and the effect on the upper airway patency. Methods: The study group included 21 patients (10 males and 11 females, mean age 8.22 ± 1.17 years) with a hyperdivergent growth and a class II malocclusion treated with AMCOP® devices. Cephalometric analysis was performed before treatment (T0) and after treatment (T1). Results: After treatment, the cephalometric analysis revealed a correction of the class II malocclusion and a modification of the growth pattern with a divergence reduction. The improvement of the upper airway space was also observed. Conclusion: The elastodontic therapy effectively corrected hyperdivergent class II malocclusion in growing patients over a short period

COVID-19 Infection in Children and Infants: Current Status on Therapies and Vaccines

Since the beginning in December 2019, the SARS-CoV-2 outbreak appeared to affect mostly the adult population, sparing the vast majority of children who only showed mild symptoms. The purpose of this investigation is to assess the status on the mechanisms that give children and infants this variation in epidemiology compared to the adult population and its impact on therapies and vaccines that are aimed towards them. A literature review, including in vitro studies, reviews, published guidelines and clinical trials was performed. Clinical trials concerned topics that allowed a descriptive synthesis to be produced. Four underlying mechanisms were found that may play a key role in providing COVID-19 protection in babies. No guidelines are available yet for therapy due to insufficient data; support therapy remains the most used. Only two vaccines are approved by the World Health Organization to be used in children from 12 years of age, and there are currently no efficacy or safety data for children below the age of 12 years. The COVID-19 clinical frame infection is milder in children and adolescents. This section of the population can act as vectors and reservoirs and play a key role in the transmission of the infection; therefore, vaccines are paramount. More evidence is required to guide safely the vaccination campaign