Micro-computed tomographic assessment of dentinal microcrack formation in straight and curved root canals in extracted teeth prepared with hand, rotary and reciprocating instruments.

AIM: To compare microcrack formation in roots of extracted teeth after the shaping of straight and curved root canals with hand, rotary and reciprocating files using micro-computed tomographic (micro-CT) analysis. METHODOLOGY: Thirty straight mandibular incisors and 30 severely curved mesial roots of mandibular molars were randomly divided into 6 experimental groups (n = 10) according to the systems used for the root canal preparation and the root canal curvature: ProTaper Universal for Hand Use (Dentsply Sirona, Ballaigues, Switzerland), HyFlex EDM (Coltene-Whaledent, Altstätten, Switzerland) and Reciproc Blue (VDW, Munich, Germany) files used in mandibular incisors (straight canals) and mesial roots of mandibular molars (curved canals). The roots were imaged with micro-CT scanning at an isotropic resolution of 14 µm before and after root canal preparation, and the cross-sectional images generated were assessed to detect microcracks. RESULTS: All dentinal defects identified after root canal preparation were already present before instrumentation, and no new microcracks were detected. Dentinal microcracks were present in 19% (ProTaper Universal for Hand Use), 11% (Hyflex EDM) and 23% (Reciproc Blue) of the cross-sections when the instrumentation was performed in mandibular incisors. Instrumentation of mandibular molars revealed microcracks in 15% (ProTaper Universal for Hand Use), 16% (Hyflex EDM) and 17% (Reciproc Blue) of the cross-sections. CONCLUSIONS: Preparation of straight and curved root canals with ProTaper Universal for Hand Use, HyFlex EDM and Reciproc Blue systems did not produce microcracks in extracted teeth when evaluated with micro-CT.

Oxidative α,ω-diyne coupling as an approach towards novel peptidic macrocycles.

The Glaser-Hay diyne coupling proved to be an efficient cyclisation approach towards diyne containing peptidic macrocycles. A variety of tetrapeptide-based macrocyclic 1,3-diynes were obtained from O-propargylated serine or tyrosine residues using Cu(OAc)2·H2O and NiCl2 under an O2-atmosphere. The effect of the linear 1,3-diyne on peptide conformations was studied by NMR and compared with a macrocycle bearing a saturated linker.

Fast and sensitive LC-MS/MS method measuring human mycotoxin exposure using biomarkers in urine.

A direct, fast and sensitive LC-MS/MS method was developed to measure biomarkers for mycotoxin exposure in human urine. In total, 32 biomarkers were quantitatively or semi-quantitatively measured in 32 urine samples of Belgian volunteers using two injections. All urine samples contained deoxynivalenol-15-glucuronide, the major detoxification metabolite of deoxynivalenol, in the ng/mL range. Also deoxynivalenol-3-glucuronide and de-epoxy-deoxynivalenol-glucuronide were present in, respectively, 90 and 25% of the samples, while deoxynivalenol was detected in 60% of the samples, in lower concentrations. Deoxynivalenol glucuronides were the major biomarkers for deoxynivalenol exposure. Ochratoxin A was detected in 70% of the samples in pg/mL. Citrinin and/or dihydrocitrinone were detected in 90% of the samples, also in concentrations of pg/mL. The presence of ochratoxin A and citrinin was confirmed by a second method using sample cleanup by immunoaffinity columns, followed by LC-MS/MS. Our data show that humans are much more exposed to citrinin than realized before and suggest further work on citrinin exposure in relation with ochratoxin A exposure, as both mycotoxins are nephrotoxic.

Seasonal variation of natural mortality factors of the guava psyllid Triozoida limbata.

It is important to understand how components of the agroecosystem interfere with the attack of a pest species and their seasonality in order to use these components in IPM programs. This study focused on the evaluation of the seasonality of natural control factors associated with the guava psyllid Triozoida limbata (Enderlein) in Brazil. Life-table data were collected from an experimental guava orchard during four periods that roughly represented four seasons. Natural mortality was monitored daily through the immature stages, and the relative importance of each natural mortality factor and its seasonality was determined. Significant statistical differences were observed in the mortality during the four periods (P < 0.05). Several factors contributed to the mortality of T. limbata, including rainfall, physiological disturbance, the parasitoid Psyllaephagus sp. (Hymenoptera: Encyrtidae) and specific predators: syrphids, predatory wasps and other generalist predators. Depending on the location of nymphs (exposed or inside galls), the relative importance of the different natural mortality factors changed. The principal component analysis (PCA) showed some trends in the relationship of natural control agents and weather conditions. For example, the occurrence of predatory wasps was positively correlated with temperature and occurrence of winds; the occurrence of syrphids and Psyllaephagus sp. were negatively correlated with temperature and winds; and the occurrence of other generalist predators were negatively correlated with the occurrence of rainfall and photoperiod. The results showed the importance of natural mortality factors for the management of T. limbata and their changes through the different seasons which should be considered when implementing IPM programs in guava orchards.

Copaiba oil and Neem extract can be a potential alternative for the behavioral control of Sitophilus zeamais.

Insects' ethology is an important factor when it is desired to carry out pest management. This knowledge makes it possible to manipulate behavioral activities, repel, or attract insects according to needs and interests. The maize weevil Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), one of the main stored grain pests, has been the target of studies of behavioral changes studies through natural substances due to its resistance to different insecticidal classes. Thus, this study aimed to evaluate the effect of sublethal concentrations of neem extract and copaiba oil on the locomotor behavior of S. zeamais. The behavioral characteristic considered were walking activity, the frequency of contact of insects with the treated grain mass, and the time spent for this behavior. The walking activity of the S. zeamais increased with exposure to Neem extract and Copaiba oil. In general, the Neem extract and Copaiba oil-induced more contact with grain mass than the control, suggesting an attractive effect on the insect, however more significant for the Neem oil. The insect's behavior was altered, presenting a specific path due to Copaiba oil and Neem extract stimuli. These results indicate that Copaiba oil and Neem extract can be a potential alternative for controlling S. zeamais on stored products since changes in this pests' behavior can reduce qualitative and quantitative grain damage. Thus, the development of products based on Copaiba oil and Neem extract may be helpful for storage pest management.

Spatiotemporal distribution of Schizaphis graminum (Rondani) and its natural enemy Coccinella septempunctata (Linnaeus) in graniferous sorghum crops.

At the global level, Sorghum bicolor (L.), is one of the most important crops, which ranks fifth among all cereals. It is cultivated in Africa, Asia, Oceania and the Americas, where it serves as a source of food and feed for humans and animals, respectively. However, production is constrained by several factors including pests and diseases. Among the pests that are known to attack Sorghum, Schizaphis graminum (Rondani), commonly called the green cereal aphid, is the most destructive pest of sorghum. S. graminum damage to sorghum is worsen by water deficit which favors its occurrence. Limited information on the spatial distribution of the pest and its natural enemies impedes the development of ecologically friendly management strategies. Therefore, the objective of this research was to determine the spatiotemporal distribution of S. graminum and its natural enemy Coccinella septempunctata (L.) during the vegetative and reproductive stage of the crop using geostatistical analysis. The spatiotemporal distribution of S. graminum was influenced by Sorghum development stage, surrounding landscape, and presence of its main predator C. septempunctata. Moreover, the abundance of C. septempunctata was influenced by the density of S. graminum. The findings from this study are required for developing sustainable pest management strategies against S. graminum.

Fungal and bacterial coinfections increase mortality of severely ill COVID-19 patients.

BACKGROUND: SARS-CoV-2 predisposes patients to secondary infections; however, a better understanding of the impact of coinfections on the outcome of hospitalized COVID-19 patients is still necessary. AIM: To analyse death risk due to coinfections in COVID-19 patients. METHODS: The odds of death of 212 severely ill COVID-19 patients were evaluated, with detailed focus on the risks for each pathogen, site of infection, comorbidities and length of hospitalization. FINDINGS: The mortality rate was 50.47%. Fungal and/or bacterial isolation occurred in 89 patients, of whom 83.14% died. Coinfected patients stayed hospitalized longer and had an increased odds of dying (odds ratio (OR): 13.45; R2 = 0.31). The risk of death was increased by bacterial (OR: 11.28) and fungal (OR: 5.97) coinfections, with increased levels of creatinine, leucocytes, urea and C-reactive protein. Coinfections increased the risk of death if patients suffered from cardiovascular disease (OR: 11.53), diabetes (OR: 6.00) or obesity (OR: 5.60) in comparison with patients with these comorbidities but without pathogen isolation. The increased risk of death was detected for coagulase-negative Staphylococcus (OR: 25.39), Candida non-albicans (OR: 11.12), S. aureus (OR: 10.72), Acinetobacter spp. (OR: 6.88), Pseudomonas spp. (OR: 4.77), and C. albicans (OR: 3.97). The high-risk sites of infection were blood, tracheal aspirate, and urine. Patients with coinfection undergoing invasive mechanical ventilation were 3.8 times more likely to die than those without positive cultures. CONCLUSION: Severe COVID-19 patients with secondary coinfections required longer hospitalization and had higher risk of death. The early diagnosis of coinfections is essential to identify high-risk patients and to determine the right interventions to reduce mortality.

Assignment and conformational investigation of asymmetric phenylindenylidene ruthenium complexes bearing N,O-bidentate ligands.

The NMR conformational study of three asymmetric phenylindenylidene ruthenium complexes 4.1-4.3, is presented. Complete (1)H and (13)C assignments could be obtained for 4.1-4.3 in benzene solution from multiple 2D homonuclear and heteronuclear NMR techniques. Our NMR analysis shows that each complex exists as a 55:45 mixture of two rotational isomers in slow exchange on the NMR chemical shift timescale. They are shown to be related by a 180 degrees flip of the indenylidene ligand along the Ru=CR bond. Both rotational isomers can be discriminated by means of NOEs contacts between the various ligands coordinating to the Ru. By matching these stereospecific assignments to the chemical shift, a chemical shift based fingerprint of the isomers that may allow straightforward assignment of future asymmetric phenylindenylidene ruthenium complexes is proposed.

Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step.

Nucleic acids are relevant biopolymers in therapy and diagnosis, for which their purity and biological activity are of crucial relevance. However, these features are difficult to achieve by cost-effective methods. Herein, we report the functionalization of a macroporous chromatographic support functionalized with an ionic liquid (IL) with remarkable performance to purify nucleic acids. An initial screening with distinct IL chemical structures supported in silica was carried out, allowing to identify the IL 1-methyl-3-propylimidazolium chloride as the most promising ligand. A chromatographic macroporous matrix able to be used in preparative liquid chromatography was then functionalized and binding/elution studies were performed. The IL 1-methyl-3-propylimidazolium chloride acts as a multimodal ligand with a remarkable dynamic binding capacity. This macroporous support allows the (one-step) purification of nucleic acids, namely small RNAs, ribosomal RNA, and genomic DNA, from a bacterial lysate, and can be regenerated and reused without compromising its separation performance.

A critical appraisal of evidence in the use of preprocedural mouthwash to avoid SARS-CoV-2 transmission during oral interventions.

OBJECTIVE: The study aimed to review and report the current evidence supporting the use of mouthwashes as a preprocedural protocol on dental offices. MATERIALS AND METHODS: This study is a secondary one that performed a comprehensive literature search of scientific studies published up to 10th August 2020 in PubMed, Scopus, Web of Science, and Scientific Electronic Library Online (Scielo) databases. The electronic search strategy was performed using free text and DeCS/MeSH terms. RESULTS: Only five studies were included in this work, despite 140 studies that were identified with the research strategy. In vivo studies were carried out in two works, in vitro studies were described in two papers, and a in silico approach was used in one work. No cetylpyridinium chloride studies were identified, while chlorhexidine and povidone studies were more studied. CONCLUSIONS: There is reduced evidence about how preprocedural mouthwashes decrease SARS-CoV-2 salivary load.

A straightforward method for quantification of vinyl functionalized water soluble alginates via 13C-NMR spectroscopy.

Alginates are fairly abundant in nature and possess many interesting properties, including their biocompatibility and ability to absorb large amounts of water. Hence, increasing interest in their derivatization has been observed and the determination of the number of newly introduced functionalities has become a key issue. For this purpose, literature generally reports on conventional 1H-NMR spectra, typically recorded at elevated temperatures and/or after hydrolysis of the alginate to circumvent line broadening effects resulting from the high viscosity. The present work reports on the modification of alginate with methacrylate functionalities and determination of the resulting degree of substitution (DS), i.e. the number of introduced methacrylate moieties relative to the initial amount of hydroxyl groups along the alginate backbone, via NMR spectroscopy. Freeze-drying and low power water presaturation were applied to improve the quality of the 1H NMR spectra. Nevertheless, it remains a qualitative method, to be used only for mutual comparisons of samples. A new and accurate method for DS determination of methacrylated alginates, based on 13C-NMR spectroscopy, is proposed. Quantitative 13C-NMR spectra were recorded with reduced measuring times by addition of a paramagnetic relaxation agent. The proposed method will also be applicable for other water-soluble functionalized alginates and polysaccharides in general.

Copaiba oil and Neem extract can be a potential alternative for the behavioral control of Sitophilus zeamais

Abstract Insects' ethology is an important factor when it is desired to carry out pest management. This knowledge makes it possible to manipulate behavioral activities, repel, or attract insects according to needs and interests. The maize weevil Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), one of the main stored grain pests, has been the target of studies of behavioral changes studies through natural substances due to its resistance to different insecticidal classes. Thus, this study aimed to evaluate the effect of sublethal concentrations of neem extract and copaiba oil on the locomotor behavior of S. zeamais. The behavioral characteristic considered were walking activity, the frequency of contact of insects with the treated grain mass, and the time spent for this behavior. The walking activity of the S. zeamais increased with exposure to Neem extract and Copaiba oil. In general, the Neem extract and Copaiba oil-induced more contact with grain mass than the control, suggesting an attractive effect on the insect, however more significant for the Neem oil. The insect's behavior was altered, presenting a specific path due to Copaiba oil and Neem extract stimuli. These results indicate that Copaiba oil and Neem extract can be a potential alternative for controlling S. zeamais on stored products since changes in this pests' behavior can reduce qualitative and quantitative grain damage. Thus, the development of products based on Copaiba oil and Neem extract may be helpful for storage pest management.

Resumo A etologia dos insetos é um fator importante quando se deseja relizar manejo de pragas. Através deste conhecimento, é possível manipular atividades comportamentais, repelir, ou atrair os insetos de acordo com as necessidades e interesses. O gorgulho do milho Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), uma das principais pragas de grãos armazenados tem sido alvo de estudos de alterações comportamentais através de substâncias naturais devido à sua resistência a diferentes classes de insecticidas. Assim, este estudo teve como objetivo avaliar o efeito de concentrações subletais de extrato de neem e do óleo de copaíba sobre o comportamento de movimentação de S. zeamais. As características comportamentais consideradas foram: a atividade de caminhamento, a frequência do contato dos insetos com a massa de grãos tratada, e o tempo gasto para realização destes comportamentos. A atividade de caminhamento do S. zeamais aumentou quando os insetos foram expostos ao extrato de Neem e ao óleo de copaíba. Em geral, o extrato de Neem e o óleo de Copaíba induziram mais contato com a massa de grãos do que o controle, sugerindo um efeito atrativo sobre o inseto, contudo este efeito foi mais significativo para o óleo de Neem. O comportamento do inseto foi alterado, apresentando um caminhamento específico devido aos estímulos do óleo de copaíba e do extrato de Neem. Estes resultados indicam que o óleo de copaíba e o extrato de Neem podem ser alternativas potenciais para o controle do S. zeamais em produtos armazenados, uma vez que alterações no comportamento desta praga podem reduzir os danos qualitativos e quantitativos nos grãos. Assim, o desenvolvimento de produtos baseados no óleo de copaíba e no extrato de Neem pode ser útil para o manejo de pragas de armazenamento.

Sn12O8(OH)4(OEt)28(HOEt)4: an additional member in the family of dodecameric oxo clusters.

A tin(IV) oxoalkoxo cluster with unprecedented architecture has been prepared and characterized by single-crystal X-ray diffraction. The cluster obeys the formula Sn 12O 8(OH) 4(OEt) 28(HOEt) 4 (1) and is based on an elongated centrosymmetric assembly of 12 six-coordinate tin centers, 28 peripheral ethoxy groups (terminal and bridging), 8 oxo bridges (mu2 and mu3), 4 hydroxy bridges (mu2), and 4 ethanol molecules that are all bound to tin atoms and interact strongly, through hydrogen bonds, with an ethoxy group located on a vicinal tin atom. This compound has also been fully characterized in solution by multinuclear 1D and 2D NMR, with all of its (119)Sn, (1)H, and (13)C NMR resonances assigned with respect to the structure. Altogether, the data allowed unambiguous location of the hydroxy groups. Information on the exchange of the ethoxy groups is also presented.

Synergistic effect of κ-carrageenan and gelatin blends towards adipose tissue engineering.

The current paper focuses on the functionalization of κ-carrageenan and gelatin as extracellular matrix polysaccharide and protein mimic respectively to produce hydrogel films for adipose tissue engineering. More specifically, κ-carrageenan as well as gelatin have been functionalized with methacrylate and methacrylamide moieties respectively to enable subsequent UV-induced crosslinking in the presence of a photo-initiator. The gel fraction, the mass swelling ratio and the mechanical properties of both the one-component hydrogels and the protein/polysaccharide blends have been evaluated. The mechanical and swelling properties of the blends could be tuned by varying the hydrogel composition as well as the crosslinking method applied. The in vitro biocompatibility assays indicated a significantly higher cell viability of adipose tissue-derived mesenchymal stem cells seeded onto the blends as compared to the one-component hydrogels. The results show that the blends of gelatin and κ-carrageenan clearly outperform the one-component hydrogels in terms of adipose tissue engineering potential.

Insecticide toxicity to the borer Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae): developmental and egg-laying effects.

Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae) is one of the major pests of solanaceous plants in South America. It is considered a great threat by the European and Mediterranean Plant Protection Organization due to the serious economic damage that it causes on tomato farms; therefore, controlling this pest is a challenging task in South America. Controlling N. elegantalis at the egg stage is the best way to prevent it from damaging crops; however, thorough studies about the effectiveness of chemicals on the different life stages of this insect pest are lacking. In this study, the effects of different chemical classes were evaluated on N. elegantalis adults, female oviposition behavior, larvae, eggs, and embryonic development. None of the tested insecticides demonstrated toxicity to the adults; however, the results showed that cartap hydrochloride affects oviposition behavior. Moreover, methomyl and cartap hydrochloride exhibited high toxicity against the eggs and larvae, with higher than 80% of mortality. These insecticides interrupted larval hatching and caused alterations in the chorion layer. Flubendiamide and deltamethrin demonstrated toxicity on N. elegantalis larvae; however, lufenuron, indoxacarb, methoxyfenozide, and chlorantraniliprole demonstrated low toxicity on both eggs and larvae, with lower than 70% of mortality. Fruit treated with cartap hydrochloride had a deterrent effect. The ovicidal activity revealed by methomyl and cartap hydrochloride might provide new approaches regarding insecticide effects on eggs. Methomyl, cartap hydrochloride, flubendiamide, and deltamethrin demonstrated toxicity on larvae. The evaluation of the chorion of the eggshell in this study has clarified the toxic effect of methomyl and cartap hydrochloride on eggs.

Modulation of hepatic glutathione transferases isoenzymes in three bivalve species exposed to purified microcystin-LR and Microcystis extracts.

This study compares the role of hepatic cytosolic glutathione transferases (cGST) isoforms of three different bivalve species to a Microcystis aeruginosa extract and purified MC-LR exposure (both at 150 µg MC-LR L-1) for 24 h. Characterization and alterations of the cytosolic GST activities in Mytilus galloprovincialis, Ruditapes philippinarum and Corbicula fluminea were measured using four class-specific substrates and changes in individual GST isoforms expression were achieved by a subsequent two-dimensional electrophoresis analysis. Evaluation of cGST activity basal levels using the four class-specific substrates denoted quantitative differences between the three bivalves. Purified MC-LR did not induce any significant response from bivalves. On the other hand, cell extracts caused significant alterations according to bivalves and substrates. Among the three bivalves, only R. philippinarum showed a significant induction of cGST activity using generic 1-chloro-2,4-dinitrobenzene (CDNB) substrate. However, no significant alterations were detected in these clams by cell extracts using the other specific substrates. In contrast, C. fluminea revealed significant induction of cGST activity when using 3,4-dichloronitrobenzene (DCNB) and ethacrynic acid (EA). In M. galloprovincialis, cell extracts promoted a significant decrease of cGST activity when using EA substrate. Altered protein expression was quantitatively detected upon exposure to cell extracts for one spot in R. philippinarum and another for C. fluminea, both upregulated (2.0 and 8.5-fold, respectively) and identified as a sigma1-class GST in the case of the first. The results showed that the three bivalves presented specific adaptive biotransformation responses to MCs and other cyanobacteria compounds supported by the modulation of distinct cGST classes.

Potential of polymethacrylate pseudo crown ethers as solid state polymer electrolytes.

The association of kinetic studies, DFT calculations and 1H-7Li NMR analyses allowed the control of the cyclo-ATRP of PEG9DMA and the production of polymethacrylate pseudo crown-ethers of various molar masses. Their potential to act as a solid-state polymer electrolyte in Li-ion batteries has been highlighted and may come from the supramolecular organization of the cyclo-PEG forming a Li+ diffusion channel.

Determination of the three-dimensional solution structure of scyllatoxin by 1H nuclear magnetic resonance.

The three-dimensional solution structure of Scyllatoxin (leiurotoxin I) a venom peptide from the scorpion Leiurus quinquestriatus hebraeus was determined at 1 A resolution by homonuclear proton n.m.r. methods at 500 MHz. Data analysis and structure calculation followed conventional protocols inherent to DIANA and related programs with two exceptions. First, distance constraints were obtained from two-dimensional nuclear Overhauser spectra by a previously described partial relaxation matrix approach. Second, since the pairing pattern of the six cysteine residues was not established a priori, the unequivocal assignment of the disulfide bridges was achieved exclusively from the n.m.r. data by a statistical analysis of preliminary DIANA structures. In the final calculation we used 227 upper distance constraints, 67 torsional constraints from vicinal coupling constants and ten stereospecific assignments of beta-methylene protons. Scyllatoxin folds into a compact ellipsoidal shape. An alpha-helix (defined with 0.24 A resolution) spanning 2.5 turns from Leu5 till Ser14 is stabilized by Cys8-Cys26 and Cys12-Cys28 disulfide bridges to the carboxy-terminal strand of an anti-parallel beta-sheet. The antiparallel beta-sheet (defined at 0.66 A resolution) extends from Leu18 to Val29 with a tight turn at Gly23-Asp24 and displays a right-handed twist. Scyllatoxin competes with the toxins apamin from Apis mellifera mellifera and P05 from Androctonus mauretanicus mauretanicus for the same or similar high conductance calcium-activated potassium channels. Consideration of presently known biological activities and three-dimensional structures of these toxins suggest a different toxin-receptor interaction of scyllatoxin as compared to apamin and P05.