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To date, studies assessing the safety profile of 3D printing materials for application in cardiac ablation are sparse. Our aim is to evaluate the safety and feasibility of two biocompatible 3D printing materials, investigating their potential use for intra-procedural guides to navigate surgical cardiac arrhythmia ablation. Herein, we 3D printed various prototypes in varying thicknesses (0.8 mm-3 mm) using a resin (MED625FLX) and a thermoplastic polyurethane elastomer (TPU95A). Geometrical testing was performed to assess the material properties pre- and post-sterilization. Furthermore, we investigated the thermal propagation behavior beneath the 3D printing materials during cryo-energy and radiofrequency ablation using an in vitro wet-lab setup. Moreover, electron microscopy and Raman spectroscopy were performed on biological tissue that had been exposed to the 3D printing materials to assess microparticle release. Post-sterilization assessments revealed that MED625FLX at thicknesses of 1 mm, 2.5 mm, and 3 mm, along with TPU95A at 1 mm and 2.5 mm, maintained geometrical integrity. Thermal analysis revealed that material type, energy source, and their factorial combination with distance from the energy source significantly influenced the temperatures beneath the 3D-printed material. Electron microscopy revealed traces of nitrogen and sulfur underneath the MED625FLX prints (1 mm, 2.5 mm) after cryo-ablation exposure. The other samples were uncontaminated. While Raman spectroscopy did not detect material release, further research is warranted to better understand these findings for application in clinical settings.

Background: The lack of thermally and mechanically performant biomaterials represents the major limit for 3D-printed surgical guides, aimed at facilitating complex surgery and ablations. Methods: Cryosurgery is a treatment for cardiac arrhythmias. It consists of obtaining cryolesions, by freezing the target tissue, resulting in selective and irreversible damage. MED625FLX and TPU95A are two biocompatible materials for surgical guides; however, there are no data on their response to cryoenergy delivery. The study purpose is to evaluate the biomaterials' thermal properties, examining the temperature changes on the porcine muscle samples (PMS) when the biomaterials are in place during the cryoablation. Two biomaterials were selected, MED625FLX and TPU95A, with two thicknesses (1.0 and 2.5 mm). To analyze the biomaterials' behavior, the PMS temperatures were measured during cryoablation, firstly without biomaterials (control) and after with the biomaterials in place. To verify the biomaterials' suitability, the temperatures under the biomaterial samples should not exceed a limit of -30.0 °C. Furthermore, the biomaterials' geometry after cryoablation was evaluated using the grid paper test. Results: TPU95A (1.0 and 2.5 mm) successfully passed all tests, making this material suitable for cryoablation treatment. MED625FLX of 1.0 mm did not retain its shape, losing its function according to the grid paper test. Further, MED625FLX of 2.5 mm is also suitable for use with a cryoenergy source. Conclusions: TPU95A (1.0 and 2.5 mm) and MED625FLX of 2.5 mm could be used in the design of surgical guides for cryoablation treatment, because of their mechanical, geometrical, and thermal properties. The positive results from the thermal tests on these materials and their thickness prompt further clinical investigation.

Background: In the field of medicine, photogrammetry has played for long time a marginal role due to the significant amount of work required that made it impractical for an extended medical use. Developments in digital photogrammetry occurred in the recent years, that have steadily increased the interest and application of this technique. The present study aims to compare photogrammetry reconstruction of heart with computed tomography (CT) as a reference. Methods: The photogrammetric reconstructions of digital images from ECG imaging derived images were performed. In particular, the ventricles of 15 patients with Brugada syndrome were reconstructed by using the free Zephyr Lite software. In order to evaluate the accuracy of the technique, measurements on the reconstructions were compared to patient-specific CT scan imported in ECG imaging software UZBCIT. Result: The results showed that digital photogrammetry in the context of ventricle reconstruction is feasible. The photogrammetric derived measurements of ventricles were not statistically different from CT scan measurements. Furthermore, the analysis showed high correlation of photogrammetry reconstructions with CT scan and a correlation coefficient close to 1. Conclusion: It is possible to reproduce digital objects by photogrammetry if the process described in this study is performed. The reconstruction of the ventricles from CT scan was very close to the values of the respective photogrammetric reconstruction., Competing Interests: ML is consultant for Atricure. GC received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Boston Scientific, Acutus Medical. CdeA receives research grants on behalf of the center from Biotronik, Medtronic, Abbott, LivaNova, Boston Scientific, AtriCure, Philips, and Acutus; CdeA received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Livanova, Boston Scientific, Atricure, Acutus Medical Daiichi Sankyo. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Talevi, Pannone, Monaco, Bori, Cappello, Candelari, Wyns, Ramak, La Meir, Gharaviri, Chierchia, Innocenti and de Asmundis.)

Background: 3D printing technology development in medical fields allows to create 3D models to assist preoperative planning and support surgical procedures. Cardiac ischemic scar is clinically associated with malignant arrhythmias. Catheter ablation is aimed at eliminating the arrhythmogenic tissue until the sinus rhythm is restored. The scope of this work is to describe the workflow for a 3D surgical guide able to define the ischemic scar and target catheter ablation., Materials and Methods: For the patient-specific 3D surgical guide and 3D heart phantom model realization, both CT scan and cardiac MRI images were processed; this was necessary to extract anatomical structures and pathological information, respectively. Medical images were uploaded and processed in 3D Slicer. For the surgical guide modeling, images from CT scan and MRI were loaded in Meshmixer and merged. For the heart phantom realization, only the CT segmentation was loaded in Meshmixer. The surgical guide was printed in MED625FLX with Polyjet technology. The heart phantom was printed in polylactide with FDM technology., Results: 3D-printed surgical model was in agreement with prespecified imputed measurements. The phantom fitting test showed high accuracy of the 3D surgical tool compared with the patient-specific reproduced heart. Anatomical references in the surgical guide ensured good stability. Ablation catheter fitting test showed high suitability of the guide for different ablation tools., Conclusion: A 3D-printed guide for ventricular tachycardia ablation is feasible and accurate in terms of measurements, stability, and geometrical structure. Concerning clinical use, further clinical investigations are eagerly awaited., Competing Interests: Author ML is consultant for AtriCure. Author GC received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Boston Scientific, Acutus Medical. Author CA received research grants on behalf of the center from Biotronik, Medtronic, Abbott, LivaNova, Boston Scientific, AtriCure, Philips, and Acutus and received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, LivaNova, Boston Scientific, AtriCure, Acutus Medical Daiichi Sankyo. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Candelari, Cappello, Pannone, Monaco, Talevi, Bori, Ramak, La Meir, Gharaviri, Chierchia, Innocenti and de Asmundis.)

Background: Brugada syndrome (BrS) is a disease associated with ventricular arrhythmias and sudden cardiac death. Epicardial ablation has demonstrated high therapeutic efficacy in preventing ventricular arrhythmias. The purpose of this research is to define a workflow to create a patient-specific 3D-printed tool to be used as a surgical guide for epicardial ablation in BrS., Methods: Due to their mechanical properties and biocompatibility, the MED625FLX and TPU95A were used for cardiac 3D surgical guide printing. ECG imaging was used to define the target region on the right ventricular outflow tract (RVOT). CT scan imaging was used to design the model based on patient anatomy. A 3D patient-specific heart phantom was also printed for fitting test. Sterilization test was finally performed., Results: 3D printed surgical models with both TPU95A and MED625FLX models were in agreement with pre-specified imputed measurements. The phantom test showed retention of shape and correct fitting of the surgical tool to the reproduced phantom anatomy, as expected, for both materials. The surgical guide adapted to both the RVOT and the left anterior descending artery. Two of the 3D models produced in MED265FLX showed damage due to the sterilization process., Conclusions: A 3D printed patient-specific surgical guide for epicardial substrate ablation in BrS is feasible if a specific workflow is followed. The design of the 3D surgical guide ensures proper fitting on the heart phantom with good stability. Further investigations for clinical use are eagerly awaited., Competing Interests: Author ML is consultant for Atricure. Author GC received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Boston Scientific, Acutus Medical. Author CA receives research grants on behalf of the center from Biotronik, Medtronic, Abbott, LivaNova, Boston Scientific, AtriCure, Philips, and Acutus; Author CA received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Livanova, Boston Scientific, Atricure, Acutus Medical Daiichi Sankyo. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Talevi, Pannone, Monaco, Bori, Cappello, Candelari, Ramak, La Meir, Gharaviri, Chierchia, Innocenti and de Asmundis.)

Background: Due to their mechanical properties, the MED625FLX and TPU95A could be appropriate candidates for cardiac 3D surgical guide use during radiofrequency ablation (RFA) treatment., Methods: RFA aims to destroy the heart tissue, which cause arrhythmias, by applying a radiofrequency (RF) energy at critical temperature above +50.0°C, where the thermal damage is considered not reversible. This study aims to analyze the biomaterials thermal properties with different thicknesses, by testing the response to bipolar and unipolar RFA on porcine muscle samples (PMS), expressed in temperature. For the materials evaluation, the tissue temperature during RFA applications was recorded, firstly without (control) and after with the biomaterials in position. The biomaterials were considered suitable for the RFA treatment if: (1) the PMS temperatures with the samples were not statistically different compared with the control; (2) the temperatures never reached the threshold; (3) no geometrical changes after RFA were observed., Results: Based on these criteria, none of the tested biomaterials resulted appropriate for unipolar RFA and the TPU95A failed almost all thermal tests also with the bipolar RFA. The 1.0 mm MED625FLX was modified by bipolar RFA in shape, losing its function. Instead, the 2.5 mm MED625FLX was considered suitable for bipolar RFA catheter use only., Conclusions: The 2.5 mm MED625FLX could be used, in the design of surgical guides for RFA bipolar catheter only, because of mechanical, geometrical, and thermal properties. None of biomaterials tested are appropriate for unipolar ablation catheter because of temperature concerns. Further investigations for clinical use are eagerly awaited., Competing Interests: Author ML is consultant for Atricure. GC received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Boston Scientific, Acutus Medical. CA receives research grants on behalf of the center from Biotronik, Medtronic, Abbott, LivaNova, Boston Scientific, AtriCure, Philips, and Acutus; received compensation for teaching purposes and proctoring from Medtronic, Abbott, Biotronik, Livanova, Boston Scientific, Atricure, Acutus Medical Daiichi Sankyo. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Cappello, Candelari, Pannone, Monaco, Bori, Talevi, Ramak, La Meir, Gharaviri, Chierchia, Innocenti and de Asmundis.)

Patient-specific three-dimensional (3D) printed models have been increasingly used in many medical fields, including cardiac surgery for which they are used as planning and communication tools. To locate and plan the correct region of interest for the bypass placement during coronary artery bypass graft (CABG) surgery, cardiac surgeons can pre-operatively rely on different medical images. This article aims to present a workflow for the production of a patient-specific 3D-printed surgical guide, from data acquisition and image segmentation to final prototyping. The aim of this surgical guide is to help visualize the region of interest for bypass placement during the operation, through the use of dedicated surgical holes. The results showed the feasibility of this surgical guide in terms of design and fitting to the phantom. Further studies are needed to assess material biocompatibility and technical properties.

Background: The number of patients presenting valgus deformities undergoing total knee arthroplasty (TKA) represents approximately 10% of the total number of TKAs performed: the presence of valgus deformity requires the implant to have proper alignment, stability and balance to achieve successful clinical outcomes, especially for knees with high coronal deformities, but these have proven to be difficult goals to achieve and therefore the use of constrained prostheses is often recommended for these cases. However, even though the use of unconstrained mobile bearing for severe knee deformities is rare, it has been shown to give successful outcomes and therefore the aim of this study is to evaluate whether this surgical technique can achieve satisfactory clinical results and correct alignment, as well as good patient satisfaction., Methods: This study presents the results of 69 TKA performed with cemented mobile bearing implants by a single surgeon on knee affected by valgus deformities. Asymmetric inserts were adopted for all the implants and an alignment surgical tool, dedicated for valgus patients, was used during the operation. Angles of valgus, WOMAC surveys and Numeric Rating Scale for pain were recorded to evaluate the results of the operations., Results: A total of 67 pre-op WOMAC questionnaire surveys were collected, with the mean result of this evaluation being 15.9 points. The Numeric Rating Scale for pain had an average of 2.2 for 68 tests. The deformities were corrected from a mean total preoperative valgus angle of 12.5° to a postoperative valgus deformity average of 0.6°. During follow-up, only one patient had serious complications due to the rupture of the extensor apparatus following a domestic accident involving falling. Further 10 patients have mild complications related to injuries such as pain of varying intensity, burning, or swelling of the knee. The level of satisfaction from 0 to 10 (0 not at all satisfied and 10 perfectly satisfied) had an average score of 7.7., Conclusions: The surgical approach presented, involving a less constrained model if compared to the ones usually chosen, allowed to achieve correct alignment and high patient satisfaction using mobile bearing implants on valgus knee deformities; further patient follow-up will be performed to evaluate long-term outcomes, but the results achieved already represent a significative finding., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 Professor P K Surendran Memorial Education Foundation. Published by Elsevier B.V. All rights reserved.)

Kimchi is recognized worldwide as the flagship food of Korea. To date, most of the currently available microbiological studies on kimchi deal with Korean manufactures. Moreover, there is a lack of knowledge on the occurrence of eumycetes in kimchi. Given these premises, the present study was aimed at investigating the bacterial and fungal dynamics occurring during the natural fermentation of an artisan non-Korean kimchi manufacture. Lactic acid bacteria were dominant, while Enterobacteriaceae, Pseudomonadaceae, and yeasts progressively decreased during fermentation. Erwinia spp., Pseudomonas veronii , Pseudomonas viridiflava , Rahnella aquatilis, and Sphingomonas spp. were detected during the first 15 days of fermentation, whereas the last fermentation phase was dominated by Leuconostoc kimchi , together with Weissella soli . For the mycobiota at the beginning of the fermentation process, Rhizoplaca and Pichia orientalis were the dominant Operational Taxonomic Units (OTUs) in batch 1, whereas in batch 2 Protomyces inundatus prevailed. In the last stage of fermentation, Saccharomyces cerevisiae , Candida sake, Penicillium , and Malassezia were the most abundant taxa in both analyzed batches. The knowledge gained in the present study represents a step forward in the description of the microbial dynamics of kimchi produced outside the region of origin using local ingredients. It will also serve as a starting point for further isolation of kimchi-adapted microorganisms to be assayed as potential starters for the manufacturing of novel vegetable preserves with high quality and functional traits.

Toxigenic and antimicrobial susceptibility patterns and genetic relatedness of 42 non-O1/O139 V. cholerae strains, the majority of them isolated from seafood and marine water of the Adriatic sea, Italy, and 9 clinical strains, two of which with seawater of the Adriatic as the source of infection, were studied. All strains had hlyA El Tor gene but lacked ctxA gene. Four and two isolates, respectively, also had stn/sto and tcpA Class genes. More than 90% of strains showed susceptibility to cefotaxime, ciprofloxacin, cloramphenicol, tetracycline, trimethoprim + sulfamethoxazole and intermediate or full resistance to tetracycline and erythromycin. Six strains of seafood and clinical source were multi-drug resistant. PFGE analysis allowed to type all the strains with 50 banding patterns. Twenty-one strains, 11 and 8 from seafood and seawater, respectively, and 2 of clinical origin, were grouped into 9 different clusters. We report the presence of toxigenic and multidrug resistant non-O1/O139 V. cholerae strains in Adriatic, some of which genetically related, and support that they represent a potential reservoir of toxin and antibiotic resistance genes., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A total of 162 samples of bivalve molluscs (45 mussels and 117 clams) collected between December 2012 and 2014 from harvesting areas of the Central Adriatic were analysed by a culturing method for the presence of Arcobacter spp. Species identification was performed by PCR and sequencing analysis of a fragment of the rpoB gene. Overall, Arcobacter species were detected in 30% of samples, specifically 33% clams and 22% mussels. A. butzleri was the most common species (20% of the samples), followed by A. cryaerophilus (9%) and A. skirrowii (1%). A seasonal association of A. butzleri contamination was detected. A. butzleri was significantly more commonly recovered from samples collected during the winter-spring period (29%) than from those of the summer-autumn (8%). A. cryaerophilus was cultured from 6% to 11% of the samples collected in summer-autumn and winter-spring, respectively, but these differences were not statistically significant. A. skirrowii was recovered from a sample of mussels harvested in May 2014. To identify associations between the occurrence of Arcobacter spp. and E. coli levels, samples were divided into groups generating results with E. coli at >230MPN/100g and E. coli at ≤230MPN/100g, the latter corresponding to EU microbiological criteria allowed for live bivalve molluscs at retail level. A. butzleri was significantly more commonly detected in samples with higher E. coli levels (48%) than in those with lower levels of E. coli (10%), providing evidence for considering E. coli as an index organism for A. butzleri contamination in bivalve molluscs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Sequencing analysis of the trh gene encoding the TDH-related haemolysin of tdh-/trh+ Vibrio parahaemolyticus isolated in Italy between 2002 and 2011 from clinical, environmental, and food samples revealed the presence of the trh2 variant in all isolates. The trh2 of the clinical isolate was 100% identical to other clinical tdh-/trh2 V. parahaemolyticus from Europe. Nucleotide and amino acid differences in the trh2 sequences of clinical isolates from Italy and other countries allowed a differentiation of the clinical strains from the majority of environmental or food strains isolated in Italy. Aspartic acid and isoleucine at positions 113 and 115, encoded by nucleotide triplets GAT and ATT at positions 337-339 and 343-345 of the complete trh gene sequence, were present in clinical strains from Europe (Italy, Norway and Germany), Asia and the United States. Only 35.5% of the tdh-/trh2 V. parahaemolyticus of environmental or food origin from Italy shared the same triplets/amino acid detected in clinical isolates, while 64.5% of isolates from the marine environment were different from those of clinical origins, demonstrating that differences occur amongst the trh2 sequences of strains from the environment and these polymorphisms may differentiate potentially pathogenic from less or non-pathogenic cultures found in the environment and seafood. In addition the distribution of T3SS2 genes was investigated in this group of tdh-/trh+ V. parahaemolyticus from different sources and in three clinical tdh+/trh- V. parahaemolyticus isolates. All tdh-/trh+ V. parahaemolyticus of environmental or food source, independent of year of isolation or geographical origin, amplified all the screened T3SS2β genes and tested negative to PCR assays for all five T3SS2α genes, as the tdh-/trh+ clinical V. parahaemolyticus isolate. The vopC genes, encoding for one of the effector proteins of T3SS2, were partially sequenced and compared to clinical tdh-/trh+ and tdh+/trh+ V. parahaemolyticus isolates from other countries. Analysis of T3SS2β vopC sequences revealed variation in tdh-/trh2 isolates from Italy, which were separated from a group of vopC sequences derived from trh2 V. parahaemolyticus from the USA., (Copyright © 2016 Elsevier B.V. All rights reserved.)

We investigated the virulence properties of four Vibrio parahaemolyticus strains causing acute gastroenteritis following consumption of indigenous mussels in Italy. The isolated strains were cytotoxic and adhesive but, surprisingly, lacked tdh, trh, and type three secretion system 2 (T3SS2) genes. We emphasize that nontoxigenic V. parahaemolyticus can induce acute gastroenteritis, highlighting the need for more investigation of the pathogenicity of this microorganism.