Acyl and CO Ligands in the [Fe]-Hydrogenase Cofactor Scramble upon Photolysis.

[Fe]-hydrogenase harbors the iron-guanylylpyridinol (FeGP) cofactor, in which the Fe(II) complex contains acyl-carbon, pyridinol-nitrogen, cysteine-thiolate and two CO as ligands. Irradiation with UV-A/blue light decomposes the FeGP cofactor to a 6-carboxymethyl-4-guanylyl-2-pyridone (GP) and other components. Previous in vitro biosynthesis experiments indicated that the acyl- and CO-ligands in the FeGP cofactor can scramble, but whether scrambling occurred during biosynthesis or photolysis was unclear. Here, we demonstrate that the [18 O1 -carboxy]-group of GP is incorporated into the FeGP cofactor by in vitro biosynthesis. MS/MS analysis of the 18 O-labeled FeGP cofactor revealed that the produced [18 O1 ]-acyl group is not exchanged with a CO ligand of the cofactor, indicating that the acyl and CO ligands are scrambled during photolysis rather than biosynthesis, which ruled out any biosynthesis mechanisms allowing acyl/CO ligands scrambling. Time-resolved infrared spectroscopy indicated that an acyl-Fe(CO)3 intermediate is formed during photolysis, in which scrambling of the CO and acyl ligands can occur. This finding also suggests that the light-excited FeGP cofactor has a higher affinity for external CO. These results contribute to our understanding of the biosynthesis and photosensitive properties of this unique H2 -activating natural complex.

[Fe]-Hydrogenase, Cofactor Biosynthesis and Engineering.

[Fe]-hydrogenase catalyzes the heterolytic cleavage of H2 and reversible hydride transfer to methenyl-tetrahydromethanopterin. The iron-guanylylpyridinol (FeGP) cofactor is the prosthetic group of this enzyme, in which mononuclear Fe(II) is ligated with a pyridinol and two CO ligands. The pyridinol ligand fixes the iron by an acyl carbon and a pyridinol nitrogen. Biosynthetic proteins for this cofactor are encoded in the hmd co-occurring (hcg) genes. The function of HcgB, HcgC, HcgD, HcgE, and HcgF was studied by using structure-to-function analysis, which is based on the crystal structure of the proteins and subsequent enzyme assays. Recently, we reported the catalytic properties of HcgA and HcgG, novel radical S-adenosyl methionine enzymes, by using an in vitro biosynthesis assay. Here, we review the properties of [Fe]-hydrogenase and the FeGP cofactor, and the biosynthesis of the FeGP cofactor. Finally, we discuss the expected engineering of [Fe]-hydrogenase and the FeGP cofactor.

Novel Digital Workflow for Nasoalveolar Molding and Postoperative Nasal Stent for Infants With Cleft Lip and Palate.

OBJECTIVE: We present a novel digital workflow to provide presurgical infant orthopedic (PSIO) treatment for a patient with a unilateral cleft lip/palate utilizing nasoalveolar molding (NAM) and a custom postsurgical nasal stent. SETTING: Within the US military healthcare system, the Joint Base San Antonio Craniofacial Anomalies Team utilizes dental scanners, predictive 3D modeling software, and 3D printing technology in a digital workflow for NAM appliance fabrication. WORKFLOW: Soft tissue facial scanning, peri-oral scanning, and dental putty impressions are used to facilitate fabrication and measure outcomes. Digital modeling software and 3D resin printing are utilized to manufacture the prescribed devices. MAIN OUTCOME MEASURES AND RESULTS: Extra-oral facial scans and intra-oral impressions are compared between 3 timepoints: pre-treatment, posttreatment with NAM, and postsurgical treatment. CONCLUSIONS: The ability to share workflows, establish outcome standards, and streamline patient care will continue to advance best practices in digital PSIO.

In Vitro Biosynthesis of the [Fe]-Hydrogenase Cofactor Verifies the Proposed Biosynthetic Precursors.

In the FeGP cofactor of [Fe]-hydrogenase, low-spin FeII is in complex with two CO ligands and a pyridinol derivative; the latter ligates the iron with a 6-acylmethyl substituent and the pyridinol nitrogen. A guanylylpyridinol derivative, 6-carboxymethyl-3,5-dimethyl-4-guanylyl-2-pyridinol (3), is produced by the decomposition of the FeGP cofactor under irradiation with UV-A/blue light and is also postulated to be a precursor of FeGP cofactor biosynthesis. HcgC and HcgB catalyze consecutive biosynthesis steps leading to 3. Here, we report an in vitro biosynthesis assay of the FeGP cofactor using the cell extract of the ΔhcgBΔhcgC strain of Methanococcus maripaludis, which does not biosynthesize 3. We chemically synthesized pyridinol precursors 1 and 2, and detected the production of the FeGP cofactor from 1, 2 and 3. These results indicated that 1, 2 and 3 are the precursors of the FeGP cofactor, and the carboxy group of 3 is converted to the acyl ligand.

The Function of Two Radical-SAM Enzymes, HcgA and HcgG, in the Biosynthesis of the [Fe]-Hydrogenase Cofactor.

In the biosynthesis of the iron-guanylylpyridinol (FeGP) cofactor, 6-carboxymethyl-5-methyl-4-hydroxy-2-pyridinol (1) is 3-methylated to form 2, then 4-guanylylated to form 3, and converted into the full cofactor. HcgA-G proteins catalyze the biosynthetic reactions. Herein, we report the function of two radical S-adenosyl methionine enzymes, HcgA and HcgG, as uncovered by in vitro complementation experiments and the use of purified enzymes. In vitro biosynthesis using the cell extract from the Methanococcus maripaludis ΔhcgA strain was complemented with HcgA or precursors 1, 2 or 3. The results suggested that HcgA catalyzes the biosynthetic reaction that forms 1. We demonstrated the formation of 1 by HcgA using the 3 kDa cell extract filtrate as the substrate. Biosynthesis in the ΔhcgG system was recovered by HcgG but not by 3, which indicated that HcgG catalyzes the reactions after the biosynthesis of 3. The data indicated that HcgG contributes to the formation of CO and completes biosynthesis of the FeGP cofactor.

Assessing the Effects of Air Abrasion with Aluminum Oxide or Glass Beads to Zirconia on the Bond Strength of Cement.

AIM: The purpose of this study was to evaluate the effects of air abrasion with aluminum oxide or glass beads to three types of zirconia containing various levels of cubic crystalline phases (3Y-TZP, Katana ML; 4Y-PSZ, Katana STML; and 5Y-PSZ, Katana UTML, Noritake) on the shear bond strength of resin cement. MATERIALS AND METHODS: Thirty block specimens (8 × 8 × 3.5 mm) were milled out of each zirconia material and mounted in plastic pipe. Ten specimens of each of the zirconia materials were air-abraded using 50 µm aluminum oxide particles, ten specimens were abraded using 80 µm glass beads, and ten specimens served as a control and received no surface treatment. A zirconia primer was applied to the surface of the zirconia specimens. Composite disks were bonded using a resin cement and light-cured. The specimens were stored in 37°C distilled water for 24 hours and thermocycled for 2,500 cycles. The specimens were loaded in shear on a universal testing machine. Data were analyzed with one-way and two-way ANOVAs and Tukey's post hoc tests (α = 0.05). RESULTS: A significant difference in shear bond strength was found based on the surface treatment (p < 0.001), but not on the type of zirconia (p = 0.132). CONCLUSION: Air abrasion with glass beads or no surface treatment resulted in significantly lower bond strength of the resin cement to all three zirconia types compared to air abrasion with aluminum oxide. CLINICAL SIGNIFICANCE: Although air abrasion with aluminum oxide may reportedly be more likely to weaken cubic-containing zirconia compared to air abrasion with glass beads, the use of aluminum oxide results in greater bond strength of the resin cement.

Assessing Flexural Strength Degradation of New Cubic Containing Zirconia Materials.

AIM: Newer zirconia materials may have greater strength degradation under cyclic fatigue with increased yttria and cubic content. The purpose of this study was to evaluate the flexural strength (FS) degradation of newer zirconia materials compared to more traditional tetragonal zirconia materials. MATERIALS AND METHODS: The following materials were tested: two 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP) materials (Lava Plus, 3M ESPE; Katana ML, Kuraray), one 4 mol% partially stabilized zirconia (4Y-PSZ) material (Katana STML, Kuraray), two 5 mol% partially stabilized zirconia (5Y-PSZ) materials (Katana STML, Kuraray; Lava Esthetic, 3M ESPE), and one lithium disilicate material (IPS e.max CAD LT, Ivoclar Vivadent). Thirty beams were milled for each ceramic material with final dimensions of 4.0 × 1.3 × 18.0 mm after sintering or crystallization. Each specimen was placed on a 3-point bend test device on a universal testing machine (Instron, Norwood, MA). Flexural strength was determined on 10 beam specimens per group with a central load applied until fracture. Flexural fatigue strength was then measured on the remaining 20 beam specimens per group using the staircase method for 6,000 cycles at 2 Hz. Data were analyzed with one-way ANOVAs/Tukey post hoc tests (α = 0.05). RESULTS: A significant difference was found between groups (p < 0.001) per property. The 3Y-TZP zirconia materials had the greatest flexural and flexural fatigue strength. The cubic containing zirconia materials performed more moderately. The lithium disilicate material had the lowest strength values. The percent degradation in flexural fatigue strength of the 3Y-TZP zirconia materials was less than the 5Y-PSZ, Katana UTML, and the 4Y-PSZ, Katana STML, cubic containing materials, but similar to the 5Y-PSZ cubic containing material, Lava Esthetic. CONCLUSION: The amount of strength degradation was material dependent, with the 4Y-PSZ or 5Y-PSZ cubic containing zirconia materials demonstrating greater or similar strength degradation compared to the primarily tetragonal 3Y-TZP zirconia materials. CLINICAL SIGNIFICANCE: The differences in FS degradation between cubic containing materials and traditional zirconia materials could significantly impact the long-term success of these newer materials. Clinicians should be aware that these cubic containing materials may perform differently long-term than the very strong traditional 3Y-TZP materials and to follow manufacturer instructions on required material thickness and indications for use to prevent premature failure of the restoration.

An Investigation of the Effect of Modifying and Reusing Impression Copings on Transfer of Implant Analog Position and Angulation.

AIM: The aim of this research was to determine whether sterilization and reutilization of impression copings had an impact on the accuracy of casts made for multiimplant restorations. MATERIALS AND METHODS: Four master casts embedded with five implant analogs were fabricated. Polyvinyl siloxane (PVS) impressions of the master cast with copings attached to the analogs were made and poured in dental stone. The impression copings were subjected to cleaning and sterilization. These processes were repeated 30 cycles for each of the two groups of five impression copings: one without modification and one with modification that included air abrasion and PVS adhesive. A coordinate measuring machine (CMM) was used to measure relative angles and distances between the reference analog and analogs. The relative angles and distances measured on the stone casts were compared to the master resin cast to obtain positional and angular displacements. RESULTS: For impression copings that were not modified, a significant difference was detected for both positional and angular displacements. For impression copings that were modified, a significant change was observed only for positional displacement. The maximum discrepancies measured for positional and angular displacements after 30 cycles of reuse were only 81 µm and 0.46°, respectively, regardless of the modification. CONCLUSION: Within the limitations of this study, unmodified impression copings that have undergone 30 cycles of cleaning and sterilization appeared to incur more impression inaccuracy than those impression copings that were modified by airborne-particle abrasion and PVS adhesive. CLINICAL SIGNIFICANCE: Impression copings used in this study can likely be recycled up to 30 times without reducing the accuracy of the impression to a level that may be considered clinically significant.

Incidence of mesorectal node metastasis in locally advanced cervical cancer: its therapeutic implications.

OBJECTIVE: To evaluate the incidence and risk factors for mesorectal node metastasis (MRNM) in locally advanced cervical cancer. METHODS/MATERIALS: We performed an observational retrospective cohort study of 122 patients with cervical cancer who received definitive chemo-radiation treatment between December 2013 and June 2017 to evaluate the incidence of MRNM. Three diagnostic radiologists assessed all available pre-treatment images. In this study, the pelvic node metastasis was defined as ≥ 1.0 cm and MRNM as ≥ 0.5 cm for CT and MRI scans and as a maximum standardized uptake value of > 2.5 for PET/CT. The relationship of MRNM with FIGO stage, pelvic node metastasis, and mesorectal fascia involvement was evaluated. RESULTS: The incidence of MRNM in all 122 patients was 8 (6.6%). However, in advanced stage (III- IV) patients, MRNM occurred in 4 of 39 (10.3%) compared with 4 of 83 (4.8%) in early stage (IB1-IIB) patients (p = 0.27). In patients with a positive pelvic node, MRNM occurred in 7 of 55 (12.7%) and 1 of 67 (1.5%) in those with negative pelvic node (p = 0.02). In addition, the incidence of MRNM was 3 of 9 (33.3%) in the presence of mesorectal fascia involvement and 5 of 113 (4.4%) among those without mesorectal fascia involvement (p = 0.013). CONCLUSION: This study indicates that pelvic node metastasis and mesorectal fascia involvement are high-risk factors for MRNM. Therefore, vigilance of reviewing images in the mesorectum for MRNM is necessary for high-risk patients.

An isotopic study of dietary diversity in formative period Ancachi/Quillagua, Atacama Desert, northern Chile.

OBJECTIVES: To characterize the paleodiet of individuals from Formative Period (1500 B.C.-A.D. 400) Atacama Desert sites of Ancachi and Quillagua as a means of understanding the dietary and cultural impacts of regional systems of exchange. MATERIALS AND METHODS: Thirty-one bone samples recovered from the cemetery of Ancachi (02QU175) and in/around the nearby town of Quillagua were the subject of carbon and nitrogen stable isotope analysis of bone collagen and hydroxyapatite and multisource mixture modeling (FRUITS, food reconstruction using isotopic transferred signals) of paleodiet. These individuals were compared with nearly 200 other Formative Period individuals from throughout the region to identify differences in dietary behaviors. RESULTS: 80.6% (25/31) of the samples yielded sufficient well-preserved collagen and were included in the multisource mixture model. The FRUITS model, which compared individuals with a robust database of available foods from the region, identified a wide diversity of diets in the Ancachi/Quillagua area (including both coastal and interior individuals), and, most notably, thirteen individuals who consumed an average of 11.2 ± 1.9% terrestrial animals, 19.8 ± 1.9% legumes, and 22.5 ± 3.1% marine fauna, a balanced pattern of protein consumption distinct from both the coastal and inland individuals in our larger regional sample. CONCLUSIONS: The combination of stable isotope analysis and multisource mixture modeling permitted the characterization of dietary behavior of 25 individuals from nodal sites in the Atacama Desert, thus enhancing our understanding of the economic and social relationships that bound together Formative Period sites, populations, and individuals in this hyperarid region.

Effect of high-speed sintering on the properties of a zirconia material.

Digital dentistry has led to the success of same-day, single-crown delivery. Unlike traditional convection furnaces for fabrication of zirconia materials, which typically require several hours to complete sintering, a dedicated high-speed sintering furnace that reportedly sinters zirconia in minutes is now available. Sintering times may increase or decrease based on the size of the restoration due to a novel interface between the chairside acquisition unit, milling unit, and furnace. The purpose of this study was to compare the flexural properties (strength and modulus), optical parameters (translucency and opalescence), and grain size of a full-contour tetragonal zirconia material after sintering in either a novel high-speed furnace or a conventional zirconia furnace. Rectangular zirconia beams were milled and sintered using the high-speed induction furnace for 26.2 minutes or a conventional convection furnace for 4.3 hours. Flexural strength and modulus were determined by subjecting the beams to a 3-point bending test in a materials testing machine. Translucency and opalescence were calculated using a spectrophotometer, and grain size was determined by using an atomic force microscope. The dedicated high-speed induction furnace sintered the tetragonal zirconia specimens in substantially less time than the conventional convection furnace, with a statistically significant decrease in grain size and no significant difference in strength or optical properties.

High-Intensity Interval Training as a Tool for Counteracting Dyslipidemia in Women.

Sedentary overweight or obese adult (age<60 years) women, allocated in type 2 diabetes mellitus (T2DM, n=13), dyslipidemia alone (DYS, n=12), dyslipidemia associated with hyperglycaemia (DYSHG, N=12), or healthy control (CON, n=10) groups, had their lipid, glucose, blood pressure, endurance performance, and anthropometry variables assessed before and after 16 weeks of a thrice-weekly high-intensity interval training (HIIT) program. Triglycerides reduced significantly (P<0.05) in all groups, and high-density lipoprotein increased (P<0.01) in T2DM, DYS and DYSHG; however, low-density lipoprotein reduced (P<0.05) only in DYSHG, and total cholesterol reduced (P<0.01) only in DYS and DYSHG. Fasting glucose reduced (P<0.05) significantly in T2DM, DYS and DYSHG, but with higher decreases in T2DM and DYSHG. Blood pressure, endurance performance and body composition improved (P<0.05) in all groups. The HIIT program was effective for restoring lipid profile of DYS and DYSHG, and fasting glucose of DYSHG to levels similar to those of CON, with a weekly time commitment 25% to 56% lower than the minimum recommended in current exercise guidelines. These findings suggest that HIIT may be a time-efficient intervention for counteracting dyslipidemia.

Fusion of Visible and Thermal Descriptors Using Genetic Algorithms for Face Recognition Systems.

The aim of this article is to present a new face recognition system based on the fusion of visible and thermal features obtained from the most current local matching descriptors by maximizing face recognition rates through the use of genetic algorithms. The article considers a comparison of the performance of the proposed fusion methodology against five current face recognition methods and classic fusion techniques used commonly in the literature. These were selected by considering their performance in face recognition. The five local matching methods and the proposed fusion methodology are evaluated using the standard visible/thermal database, the Equinox database, along with a new database, the PUCV-VTF, designed for visible-thermal studies in face recognition and described for the first time in this work. The latter is created considering visible and thermal image sensors with different real-world conditions, such as variations in illumination, facial expression, pose, occlusion, etc. The main conclusions of this article are that two variants of the proposed fusion methodology surpass current face recognition methods and the classic fusion techniques reported in the literature, attaining recognition rates of over 97% and 99% for the Equinox and PUCV-VTF databases, respectively. The fusion methodology is very robust to illumination and expression changes, as it combines thermal and visible information efficiently by using genetic algorithms, thus allowing it to choose optimal face areas where one spectrum is more representative than the other.

Effect of Vertical, Horizontal, and Combined Plyometric Training on Explosive, Balance, and Endurance Performance of Young Soccer Players.

The aim of this study was to compare the effects of 6 weeks of vertical, horizontal, or combined vertical and horizontal plyometric training on muscle explosive, endurance, and balance performance. Forty young soccer players aged between 10 and 14 years were randomly divided into control (CG; n = 10), vertical plyometric group (VG; n = 10), horizontal plyometric group (HG; n = 10), and combined vertical and horizontal plyometric group (VHG; n = 10). Players performance in the vertical and horizontal countermovement jump with arms, 5 multiple bounds test (MB5), 20-cm drop jump reactive strength index (RSI20), maximal kicking velocity (MKV), sprint, change of direction speed (CODS), Yo-Yo intermittent recovery level 1 test (Yo-Yo IR1), and balance was measured. No significant or meaningful changes in the CG, apart from small change in the Yo-Yo IR1, were observed while all training programs resulted in meaningful changes in explosive, endurance, and balance performance. However, only VHG showed a statistically significant (p ≤ 0.05) increase in all performance test and most meaningful training effect difference with the CG across tests. Although no significant differences in performance changes were observed between experimental groups, the VHG program was more effective compared with VG (i.e., jumps, MKV, sprint, CODS, and balance performance) and HG (i.e., sprint, CODS, and balance performance) to small effect. The study demonstrated that vertical, horizontal, and combined vertical and horizontal jumps induced meaningful improvement in explosive actions, balance, and intermittent endurance capacity. However, combining vertical and horizontal drills seems more advantageous to induce greater performance improvements.

Metagenome-assembled bacterial genomes recovered from the datasets of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae).

Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), also known as the fall armyworm, is an economically important and widespread polyphagous pest. Microorganisms associated to this insect during life cycle play important ecological roles. We report 3 metagenome-assembled bacterial genomes reconstructed from a metagenome dataset obtained from S. frugiperda larvae F3 3rd-instar reared using artificial diet under laboratory conditions. Genome data for Enterococcus casseliflavus indicated a genome length of 3,659,8333 bp and GC content of 42.54%. Genome data for E. mundtii indicated a genome length of 2,921,701 bp and GC content of 38.37%. Finally, genome data for Lactiplantibacillus plantarum indicated a genome length of 3,298,601 bp, GC content of 44.31%. Genome analysis allowed us to identify genus-specific protein families (PLFams), transporters and antibiotic resistance-related genes among others. DNA sequences were deposited in National Center for Biotechnology Information (https://www.ncbi.nlm.nih.gov/) as Bioproject accession PRJNA899064.

Splinting or non-splinting of fixed prostheses on adjacent implants: A critical review.

PURPOSE: The present study aimed to identify, through a critical review of the literature, the success factors associated with the splinting of fixed prostheses on adjacent implants of the posterior sectors in partially edentulous patients compared with those not splinted. STUDY SELECTION: A MEDLINE strategy was implemented based on a research question to systematically search and extract information from databases (PubMed and Scopus) using MeSH terms/keywords identified for each domain. Systematic reviews, clinical and in vitro studies were selected and classified according to eligibility criteria based on the research question and level of evidence using the PRISMA flowchart. RESULTS: A total of 32 studies were selected for data extraction and analysis according to study design (three systematic reviews, 14 clinical studies, and 15 in vitro studies). Overall, the studies found no significant difference in the association between the survival rate and prosthesis type. In clinical studies, there have been no differences in marginal bone loss between splinted and non-splinted prostheses, and the influence of peri-implant status and restorative materials has been poorly evaluated. The distribution of stress and loads determined in the in vitro studies showed results that could favor splinted prostheses; however, are generally associated with implant design. CONCLUSIONS: The need for splinted or non-splinted adjacent implant-supported prostheses remains controversial. The reviewed evidence indicates that factors such as implant size and its relationship with coronal height could be important in decision-making.

Kosakonia cowanii Ch1 Isolated from Mexican Chili Powder Reveals Growth Inhibition of Phytopathogenic Fungi.

Kosakonia cowanii strain Ch1 was isolated from Mexican chili powder, and the genome was sequenced. The genome was 4,765,544 bp in length, with an average G + C content of 56.22%, and a plasmid (pCh1) of 128,063 bp with an average G + C content of 52.50%. A phylogenetic analysis revealed a close relation with pathogenic strains; nevertheless, some virulence-related genes were absent, and this genetic characteristic may explain the fact that K. cowanii Ch1 behaved as a non-pathogenic strain when infection assays were performed on the leaves and fruits of Capsicum annuum L. Surprisingly, we observed that this bacterial strain had the ability to spread throughout serrano pepper seeds. Furthermore, K. cowanii Ch1 was evaluated for the production of volatile organic compounds (VOCs) against fungal pathogens, and the results showed that Alternaria alternata and Sclerotium rolfsii were inhibited in a radial mycelial growth assay by a mean rate of 70% and 64%, while Fusarium oxysporum was inhibited by only approximately 10%. Based on the headspace solid-phase microextraction combined with the gas chromatography mass spectrometry (HS-SPME-GC-MS), 67 potential VOCs were identified during the fermentation of K. cowanii Ch1 in TSA medium. From these VOCs, nine main compounds were identified based on relative peak area: dodecanoic acid; 3-hydroxy ethanol; 1-butanol-3-methyl; acetaldehyde; butanoic acid, butyl ester; cyclodecane; 2-butanone, 3-hydroxy; disulfide, dimethyl and pyrazine-2,5-dimethyl. Our findings show the potential of K. cowanii Ch1 for the biocontrol of fungal pathogens through VOCs production and reveal additional abilities and metabolic features as beneficial bacterial specie.

Detection of Bacillus cereus sensu lato Isolates Posing Potential Health Risks in Mexican Chili Powder.

The potential presence of spore-forming bacteria related to the Bacillus cereus group in Mexican chili powder elaborated from Capsicum annuum L. is of commercial and clinical interest, because chili powder is an essential spice in the Mexican diet and in diets around the globe. To facilitate detection and isolation of members of this group of spore-forming bacteria from Mexican chili powder samples, we identified colonies that grew on agar medium selective for Bacillus cereus sensu lato, supplemented with polymyxin B (10 µg/mL) and ampicillin (10 to 100 µg/mL). The presumptive B. cereus (s.l.) isolates were tested using a tRNACys-PCR-based approach and the results identified species related phylogenetically to B. cereus, B. thuringiensis, and B. toyonensis. Their toxigenic potential was assessed by serological tests to detect enterotoxins (Nhe and Hbl) and by PCR targeting the hemolysin BL (hbl) component C (hblC) and non-hemolytic enterotoxin component A (nheA). The antibiotic profiles of the isolates showed a high resistance to ß-lactams (100% of the isolates), trimethoprim-sulfamethoxazole (100%), tetracycline (90%), erythromycin (77%), clindamycin (74%), and chloramphenicol (42%). Our results indicate the presence of B. cereus s.l. with toxigenic characteristics in Mexican chili powder. Because of the potential for these organisms to cause disease through their production of various toxins, and resistance to antibiotics, we recommend that a microbiological risk assessment must be considered in the Mexican regulatory requirements.

Complete Real-Scale Application of Recycled Aggregates in a Port Loading Platform in Huelva, Spain.

The application of recycled aggregates (RA) from construction and demolition waste and crushed concrete blocks is a very important challenge for the coming years from the environmental point of view, in order to reduce the exploitation of natural resources. In Spain, the use of these recycled materials in the construction of road bases and sub-bases is growing significantly. However, presently, there are few studies focused on the properties and behavior of RA in civil works such as road sections or seaport platforms. In this work, two types of RA were studied and used in a complete real-scale application. Firstly, recycled concrete aggregates (RCA) were applied in the granular base layer under bituminous superficial layers, and secondly mixed recycled aggregates (MRA) which contain a mix of ceramic, asphalt, and concrete particles were applied in the granular subbase layer, under the base layer made with RCA. Both RA were applied in a port loading platform in Huelva, applying a 100% recycling rate. This civil engineering work complied with the technical requirements of the current Spanish legislation required for the use of conventional aggregates. The environmental benefits of this work have been very relevant, and it should encourage the application of MRA and RCA in civil engineering works such as port platforms in a much more extended way. This is the first and documented real-scale application of RA to completely build the base and sub-base of a platform in the Huelva Port, Spain, replacing 100% of natural aggregates with recycled ones.