The dangers of reused personal protective equipment: healthcare workers and workstation contamination.

BACKGROUND: Personal protective equipment (PPE) is essential to protect healthcare workers (HCWs). The practice of reusing PPE poses high levels of risk for accidental contamination by HCWs. Scarce medical literature compares practical means or methods for safe reuse of PPE while actively caring for patients. METHODS: In this study, observations were made of 28 experienced clinical participants performing five donning and doffing encounters while performing simulated full evaluations of patients with coronavirus disease 2019. Participants' N95 respirators were coated with a fluorescent dye to evaluate any accidental fomite transfer that occurred during PPE donning and doffing. Participants were evaluated using blacklight after each doffing encounter to evaluate new contamination sites, and were assessed for the cumulative surface area that occurred due to PPE doffing. Additionally, participants' workstations were evaluated for contamination. RESULTS: All participants experienced some contamination on their upper extremities, neck and face. The highest cumulative area of fomite transfer risk was associated with the hook and paper bag storage methods, and the least contamination occurred with the tabletop storage method. Storing a reused N95 respirator on a tabletop was found to be a safer alternative than the current recommendation of the US Centers for Disease Control and Prevention to use a paper bag for storage. All participants donning and doffing PPE were contaminated. CONCLUSION: PPE reusage practices pose an unacceptably high level of risk of accidental cross-infection contamination to healthcare workers. The current design of PPE requires complete redesign with improved engineering and usability to protect healthcare workers.

Color Change of Resin-based Composites After In Vitro Bleaching Protocols: A Systematic Review and Meta-analysis.

OBJECTIVES: To systematically review the literature on color stability of resin-based composites (RBC) after in vitro bleaching protocols and to assess the influence of bleaching protocols by meta-regression analysis on RBC color stability, and the association with clinical and experimental characteristics. METHODS: The electronic search was conducted in MEDLINE/PubMed, Scopus, and Web of Science databases and included English language studies that evaluated and reported color differences (CIELAB values) of RBC after in vitro bleaching procedures using hydrogen peroxide and/or carbamide peroxide. RESULTS: Database search for color change of RBC after bleaching retrieved 1335 eligible papers after removing duplicates. After initial screening, 66 articles were assessed for full-text reading with final inclusion of 23 published papers. A meta-regression analysis showed that storage time (p≤0.01), color measuring device (p≤0.01), and background color (p≤0.01) had influenced on color changes of RBC. The bleaching protocol (bleaching agent and time of application) did not influence on color changes of RBC (p>0.01). CONCLUSIONS: There is evidence that RBC change color after bleaching, but the change is not clinically significant.

Orogastric intubation with dental fixation for enteral nutrition: a proof-of-concept study.

In this crossover trial, we evaluated a new technique for enteral nutrition using orogastric intubation. Twelve volunteers were randomly assigned to both orogastric (OGI) and conventional nasogastric intubation (NGI) with a 15-day interval. The tip of the orogastric tube was fixed intraorally into an upper molar. Participants were asked to remain intubated for 24 hours and rated efficacy and safety using a Likert scale (1: worst / 10: best). Tolerance in hours was longer during OGI (median 21 versus 12.5; p=0.022). OGI was superior in comfort (median 7 versus 3; p=0.002), aesthetic (median 10 versus 1; p=0.002), speech (median 5.5 versus 3; p=0.014) and swallowing (median 8 versus 2; p=0.004). Both procedures were tolerated with mild local complaints. Diet volume through the tube was greater during NGI (p = 0.014). In healthy participants, orogastric intubation with dental fixation showed greater efficacy and similar safety to nasogastric intubation. CLINICAL TRIALS NUMBER: NCT03670238.

Dyslipidaemia after switch to tenofovir alafenamide (TAF)-based cART regimens in a cohort of HIV-positive patients: what clinical relevance?

OBJECTIVES: Switching from tenofovir (TDF) to tenofovir alafenamide (TAF) affects lipid profile. The aim of this study was to evaluate whether this results in an increased frequency of patients with low-density lipoprotein (LDL) above their cardiovascular-related target. METHODS: All HIV patients switching from TDF to TAF, with no changes of the anchor drug, and with plasma lipids available within 6 months before and after the switch, were included. Demographic, HIV-related parameters, cardiovascular (CV) risk factors and lipid profile on both TDF and TAF were collected. The CV risk score and the relative target of LDL for each patient were calculated according to 2016 European Society of Cardiology/European Atherosclerosis Society (ESC/EAS) guidelines for the management of dyslipidaemias. Modifications in lipid profiles and in the prevalence of patients with LDL above their CV-related target were evaluated after switch to TAF. RESULTS: Overall, 221 HIV patients were included, according to CV risk: 55% at low risk, 34% at moderate risk, and 11% at high/very high risk. By analysing lipid profiles according to CV risk, 38% of patients on TDF had LDL above their CV target; this prevalence increases to 60% after switching to TAF (P < 0.0001). The presence of cobicistat in the combination antiretroviral therapy (cART) regimen was associated with an increased risk of LDL above the CV-related target after switch to TAF [adjusted odds ratio (aOR) = 2.4, 95% confidence interval (CI): 1-5.1], P = 0.03) and with an increased prescription of lifestyle/therapeutic intervention (OR = 3.0, 95% CI: 1.7-5.3, P < 0.0001). DISCUSSION: Switching from TDF to TAF affects lipid parameters, and data from real life suggest a clinical relevance of this worsening that often leads clinicians to implement lifestyle/therapeutic interventions.

Influence of Bleaching and Aging Procedures on Color and Whiteness of Dental Composites.

Bleaching can cause perceptible color changes on resin-based composite (RBC) restorations that may not be stable with aging. The objective of this study was to evaluate color stability and whiteness variations of RBCs after bleaching and aging procedures. Discs (10 mm in diameter and 1 mm thick) of shades A2 and A3 were fabricated from two RBCs (Filtek Z250 and Filtek Z350 XT) and divided into three subgroups (for each composite and shade) (n=5) as follows: control (no bleaching), at-home bleaching, and in-office bleaching. All specimens underwent an accelerated artificial aging up to 450 KJ/m2 and 900 KJ/m2 in an aging chamber (Suntest XXL+). A spectroradiometer (SpectraScan PR-670) was used to obtain CIE L*a*b* coordinates. CIEDE2000 color difference (ΔE00) and whiteness index for dentistry (WID) were used to evaluate color stability. Color and whiteness differences data were analyzed considering the 50:50% visual color difference thresholds (perceptibility [PT] and acceptability [AT]) and 50:50% whiteness thresholds (whiteness perceptibility [WPT] and whiteness acceptability [WAT]). Analysis of variance and Tukey tests (α=0.05) were used to statistically analyze the data. After bleaching, all specimens showed ΔE00 and ΔWID values below their corresponding acceptability thresholds (AT and WAT, respectively). After aging, L* and WID values decreased while b* values increased (p≤0.05), resulting in ΔE00 and ΔWID values above AT and WAT, respectively. Color changes after bleaching RBCs were clinically acceptable, while aging provoked clinically perceptible color changes.

Spectroscopic identification of the chemical interplay between defects and dopants in Al-doped ZnO.

The conduction and optoelectronic properties of transparent conductive oxides can be largely modified by intentional inclusion of dopants over a very large range of concentrations. However, the simultaneous presence of structural defects results in an unpredictable complexity that prevents a clear identification of chemical and structural properties of the final samples. By exploiting the unique chemical sensitivity of Hard X-ray Photoelectron Spectra and Near Edge X-ray Absorption Fine Structure in combination with Density Functional Theory, we determine the contribution to the spectroscopic response of defects in Al-doped ZnO films. Satellite peaks in O1s and modifications at the O K-edge allow the determination of the presence of H embedded in ZnO and the very low concentration of Zn vacancies and O interstitials in undoped ZnO. Contributions coming from substitutional and (above the solubility limit) interstitial Al atoms have been clearly identified and have been related to changes in the oxide stoichiometry and increased oxygen coordination, together with small lattice distortions. In this way defects and doping in oxide films can be controlled, in order to tune their properties and improve their performances.

ADM guidance-ceramics: Fatigue principles and testing.

BACKGROUND: Clinical failure of dental ceramics is usually reported as partial fracture of the restoration (chipping) or as catastrophic fracture of the whole structure. In contrast to metals, ceramics are linear-elastic, brittle materials exhibiting extremely low damage tolerance to failure. Well documented clinical and lab reports have shown this fracture event often occurs at loads far below their fracture strength due to intrinsic fatigue degradation via slow crack growth or cyclic fatigue mechanisms. The presence and development of surface flaws have a dominant role in damage accumulation and lifetime reduction of ceramic structures. AIMS: This ADM guidance document aims to summarize the aspects related to fatigue degradation of dental ceramics, reviewing the concepts of fatigue testing and furthermore aims to provide practical guidance to young scientists entering into fatigue related research. The description of fatigue strength is always accompanied by a clear understanding of the underlying fracture mechanisms.

Marijuana smoke induces severe pulmonary hyperresponsiveness, inflammation, and emphysema in a predictive mouse model not via CB1 receptor activation.

Sporadic clinical reports suggested that marijuana smoking induces spontaneous pneumothorax, but no animal models were available to validate these observations and to study the underlying mechanisms. Therefore, we performed a systematic study in CD1 mice as a predictive animal model and assessed the pathophysiological alterations in response to 4-mo-long whole body marijuana smoke with integrative methodologies in comparison with tobacco smoke. Bronchial responsiveness was measured with unrestrained whole body plethysmography, cell profile in the bronchoalveolar lavage fluid with flow cytometry, myeloperoxidase activity with spectrophotometry, inflammatory cytokines with ELISA, and histopathological alterations with light microscopy. Daily marijuana inhalation evoked severe bronchial hyperreactivity after a week. Characteristic perivascular/peribronchial edema, atelectasis, apical emphysema, and neutrophil and macrophage infiltration developed after 1 mo of marijuana smoking; lymphocyte accumulation after 2 mo; macrophage-like giant cells, irregular or destroyed bronchial mucosa, goblet cell hyperplasia after 3 mo; and severe atelectasis, emphysema, obstructed or damaged bronchioles, and endothelial proliferation at 4 mo. Myeloperoxidase activity, inflammatory cell, and cytokine profile correlated with these changes. Airway hyperresponsiveness and inflammation were not altered in mice lacking the CB1 cannabinoid receptor. In comparison, tobacco smoke induced hyperresponsiveness after 2 mo and significantly later caused inflammatory cell infiltration/activation with only mild emphysema. We provide the first systematic and comparative experimental evidence that marijuana causes severe airway hyperresponsiveness, inflammation, tissue destruction, and emphysema, which are not mediated by the CB1 receptor.

Masking Colored Substrates Using Monolithic and Bilayer CAD-CAM Ceramic Structures.

OBJECTIVE: To evaluate the masking ability and translucency of monolithic and bilayer CAD-CAM ceramic structures. METHODS: Discs of high translucency (HT) and low translucency (LT) lithium disilicate-based ceramic (IPS e.max CAD) with different thicknesses (0.7, 1, 1.5, and 2 mm) were evaluated as a monolithic structure or combined (bilayer) with a 0.5-mm-thick zirconia framework (IPS e.max ZirCAD). The masking ability and translucency were calculated based on CIE L*a*b* color coordinates measured with a spectrophotometer (SP60, X-Rite). The translucency parameter (TP) was calculated using color coordinates measured over standard white-and-black backgrounds. The masking ability was calculated by CIEDE2000 color difference metric (ΔE00) for each specimen measured over a tooth-colored substrate (shade A2) compared to three darker backgrounds (shade C4 and two metal substrates). Confidence intervals (CI) for the means (95% CI) were calculated for TP and ΔE00. The Pearson correlation between ΔE00 and TP was investigated for monolithic and bilayer structures over all backgrounds. RESULTS: The thinner the lithium disilicate layer, the greater the translucency and the higher the ΔE00 values. The effect of ceramic thickness on both translucency and masking ability was more pronounced for the monolithic structures. In addition, monolayers always presented a greater color variation than their bilayer counterparts. The metallic background produced greater ΔE00 than the C4-shaded substrate. CONCLUSION: Monolithic veneers were able to mask C4-shaded background but did not mask metallic backgrounds. Bilayer structures showed greater shade masking ability than monolithic structures.

Reliability and failure behavior of CAD-on fixed partial dentures.

OBJECTIVES: To estimate the reliability and failure behavior of fixed partial dentures (FPDs) fabricated using the CAD-on technique. METHODS: FPDs (n=25) were fabricated using a CAD/CAM system: IPS e.max ZirCAD - Crystall./Connect and IPS e.max CAD (Ivoclar). The restoration type ("three-unit bridge") and design method ("multilayer") based on Biogenerics were used. Framework and porcelain structures were united using a fusion ceramic (Crystall./Connect, Ivoclar). Mechanical fatigue was tested in a servohydraulic load frame machine at a cyclic loading (frequency: 2Hz; load ratio: 0.1). Based on previous data from specimens tested in fast fracture, three different stress profiles were used. The lifetime data were analyzed using an inverse power law-Weibull cumulative damage model (ALTA PRO, Reliasoft). All failed specimens were examined under a field emission scanning electron microscope. RESULTS: Porcelain chipping was the predominant (60%) mode of failure for FPDs tested in fast fracture and connector failure was predominant (67%) under fatigue. For fast fracture data, the Weibull modulus (ß) of FPDs was 7.8 combining the two failure modes. When chipping and connector fracture data were analyzed separately, ß values were 7.9 and 2.9. For the step stress fatigue test, ß values were lower than estimated using fast fracture, being 1.6 for connector fracture and 1.3 for porcelain chipping. SIGNIFICANCE: The test method (fast fracture or fatigue) significantly influenced the reliability of FPDs fabricated using the CAD-on technique, but it did not influence their failure behavior.

Flexural strength and reliability of monolithic and trilayer ceramic structures obtained by the CAD-on technique.

OBJECTIVE: To evaluate the flexural strength, Weibull modulus, fracture toughness, and failure behavior of ceramic structures obtained by the CAD-on technique, testing the null hypothesis that trilayer structures show similar properties to monolithic structures. METHODS: Bar-shaped (1.8mm×4mm×16mm) monolithic specimens of zirconia (IPS e.max ZirCAD - Ivoclar Vivadent) and trilayer specimens of zirconia/fusion ceramic/lithium dissilicate (IPS e.max ZirCAD/IPS e.max CAD Crystall./Connect/IPS e.max CAD, Ivoclar Vivadent) were fabricated (n=30). Specimens were tested in flexure in 37°C deionized water using a universal testing machine at a crosshead speed of 0.5mm/min. Failure loads were recorded, and the flexural strength values were calculated. Fractography principles were used to examine the fracture surfaces under optical and scanning electron microscopy. Data were statistically analyzed using Student's t-test and Weibull statistics (α=0.05). RESULTS: Monolithic and trilayer specimens showed similar mean flexural strengths, characteristic strengths, and Weibull moduli. Trilayer structures showed greater mean critical flaw and fracture toughness values than monolithic specimens (p<0.001). Most critical flaws in the trilayer groups were located on the Y-TZP surface subjected to tension and propagated catastrophically. Trilayer structures showed no flaw deflection at the interface. SIGNIFICANCE: Considering the CAD-on technique, the trilayer structures showed greater fracture toughness than the monolithic zirconia specimens.

Lifetime comparison of Y-TZP/porcelain crowns under different loading conditions.

OBJECTIVES: To compare the lifetime of Y-TZP/porcelain crowns under three different load conditions using step-stress accelerated lifetime testing. METHODS: The Y-TZP frameworks were milled using CAD/CAM, veneered with a porcelain and cemented onto dentine analogue dies. Specimens were divided according to the occlusal load condition (n=20): central fossa load (CFL), cusp tip load (CTL) and sliding contact (SC). For CFL and CTL, the cyclic load was applied parallel to the long axis of the preparation using a ceramic piston. For SC, the axial load was associated to 1mm lateral displacement at the disto-lingual cusp. Different stress profiles were used. Failures were detected with an acoustic system. A Weibull distribution (95% confidence boundary) was used to analyse the data, and fractographic principles were used to evaluate fractured specimens. RESULTS: The acoustic monitor was able to detect the initial crack. The probability of failure (at 300 N load and 200,000 cycles) was statistically greater for CTL (0.63; 0.44-0.81) compared to CFL (0.23; 0.12-0.43). The Weibull modulus of CFL (2.1; 1.5-3.6) was greater than for SC (0.7; 0.5-1.2), with no difference in the lifetime. All specimens failed by chipping, which originated predominantly at the contact (66.7%) on CTL, and in the bulk of the porcelain on CFL (100%) and SC (80%). CONCLUSIONS: Contact at the cusp tip is more harmful than at the central fossa. Data from sliding contact are less consistent than from axial contacts, but more clinically relevant. CLINICAL SIGNIFICANCE: The loading condition of Y-TZP/porcelain crowns can influence on the probability of failure and failure mode. The contact at the cusp tip is more harmful than at the central fossa, where the stress is better distributed. Sliding contact is a better simulator of the chewing cycle compared to axial contacts.

Multiple Orientia tsutsugamushi ankyrin repeat proteins interact with SCF1 ubiquitin ligase complex and eukaryotic elongation factor 1 α.

BACKGROUND: Orientia tsutsugamushi, the causative agent of scrub typhus, is an obligate intracellular bacterium. Previously, a large number of genes that encode proteins containing eukaryotic protein-protein interaction motifs such as ankyrin-repeat (Ank) domains were identified in the O. tsutsugamushi genome. However, little is known about the Ank protein function in O. tsutsugamushi. METHODOLOGY/PRINCIPAL FINDINGS: To characterize the function of Ank proteins, we investigated a group of Ank proteins containing an F-box-like domain in the C-terminus in addition to the Ank domains. All nine selected ank genes were expressed at the transcriptional level in host cells infected with O. tsutsugamushi, and specific antibody responses against three Ank proteins were detected in the serum from human patients, indicating an active expression of the bacterial Ank proteins post infection. When ectopically expressed in HeLa cells, the Ank proteins of O. tsutsugamushi were consistently found in the nucleus and/or cytoplasm. In GST pull-down assays, multiple Ank proteins specifically interacted with Cullin1 and Skp1, core components of the SCF1 ubiquitin ligase complex, as well as the eukaryotic elongation factor 1 α (EF1α). Moreover, one Ank protein co-localized with the identified host targets and induced downregulation of EF1α potentially via enhanced ubiquitination. The downregulation of EF1α was observed consistently in diverse host cell types infected with O. tsutsugamushi. CONCLUSION/SIGNIFICANCE: These results suggest that conserved targeting and subsequent degradation of EF1α by multiple O. tsutsugamushi Ank proteins could be a novel bacterial strategy for replication and/or pathogenesis during mammalian host infection.

Controlled co-deposition of FePt nanoparticles embedded in MgO: a detailed investigation of structure and electronic and magnetic properties.

Films of FePt nanoparticles (NPs) embedded in MgO were obtained by controlled co-deposition of FePt NPs pre-formed by a gas aggregation source and of Mg evaporated in an oxygen atmosphere. Assemblies of core-shell FePt@MgO NPs and films of FePt NPs embedded in MgO matrix could be obtained by varying FePt and Mg deposition rates. Transmission electron microscopy (TEM) and high resolution-TEM revealed the core-shell structure of the NPs, with an FePt core (of average diameter (d) = 4.75 nm) presenting a multitwinned icosahedral structure, and MgO partially in crystalline form. The functional effect of the MgO shell in shielding the FePt core from external oxidation was shown with XPS. Upon controlled annealing, a transition from A1 to L10 ordering could be obtained, with structural and morphological re-arrangement. The magnetic hysteresis loops obtained from alternating gradient field magnetometry at room temperature show a 'wasp-waist' shape, with small values of coercive field (Hc = 300-1400 Oe), decreasing at increasing amounts of co-deposited MgO.

Phenotypic characterization of peripheral T cells and their dynamics in scrub typhus patients.

BACKGROUND: Scrub typhus, caused by Orientia tsutsugamushi infection, is one of the main causes of febrile illness in the Asia-Pacific region. Although cell-mediated immunity plays an important role in protection, little is known about the phenotypic changes and dynamics of leukocytes in scrub typhus patients. METHODOLOGY/PRINCIPAL FINDINGS: To reveal the underlying mechanisms of immunological pathogenesis, we extensively analyzed peripheral blood leukocytes, especially T cells, during acute and convalescent phases of infection in human patients and compared with healthy volunteers. We observed neutrophilia and CD4(+) T lymphopenia in the acute phase of infection, followed by proliferation of CD8(+) T cells during the convalescent phase. Massive T cell apoptosis was detected in the acute phase and preferential increase of CD8(+) T cells with activated phenotypes was observed in both acute and convalescent phases, which might be associated or correlated with elevated serum IL-7 and IL-15. Interestingly, peripheral Treg cells were significantly down-regulated throughout the disease course. CONCLUSIONS/SIGNIFICANCE: The remarkable decrease of CD4(+) T cells, including Treg cells, during the acute phase of infection may contribute to the loss of immunological memory that are often observed in vaccine studies and recurrent human infection.

IL-7Rαlow memory CD8+ T cells are significantly elevated in patients with systemic lupus erythematosus.

OBJECTIVE: Human effector memory (EM) CD8(+) T cells include IL-7Rα(high) and IL-7Rα(low) cells with distinct cellular characteristics, including the expression of cytotoxic molecules. Both NK cells and the NK cell-associated molecule 2B4 that is expressed on CD8(+) T cells promote cytotoxicity. Here we analysed the expression of 2B4 on IL-7Rα(high) and IL-7Rα(low) EM CD8(+) T cells and its contribution to cytotoxicity. We also analysed the frequency of IL-7Rα(high) and IL-7Rα(low) EM CD8(+) T cells in patients with SLE or lupus and in healthy individuals given the potential role of cytotoxic CD8(+) T cells in the pathogenesis of lupus. METHODS: We used flow cytometry to measure the expression of 2B4 on IL-7Rα(high) and IL-7Rα(low) EM CD8(+) T cells as well as the frequency of these cell populations in the peripheral blood of healthy individuals and patients with SLE. Also, 2B4-mediated cytotoxicity was quantitated in IL-7Rα(high) and IL-7Rα(low) EM CD8(+) T cells using target cells with CD48 antigen. RESULTS: We found that IL-7Rα(high) EM CD8(+) T cells had higher levels of 2B4 expression compared with IL-7Rα(low) EM CD8(+) T cells. Triggering 2B4 enhanced the cytotoxic function of IL-7Rα(low) EM CD8(+) T cells against target cells. We also noticed that patients with SLE had an increased frequency of IL-7Rα(low) EM CD8(+) T cells that correlated with disease manifestation. CONCLUSION: Our findings show that SLE patients have increased IL-7Rα(low) EM CD8(+) T cells, possibly contributing to tissue damage through 2B4-mediated cytotoxicity.

Characterization of effector memory CD8+ T cells in the synovial fluid of rheumatoid arthritis.

Little is known about the cellular characteristics of CD8(+) T cells in rheumatoid arthritis (RA). We addressed this by investigating whether the frequency of the CD8(+) T cell subsets and their phenotypic characteristics are altered in the peripheral blood and synovial fluid (SF) from patients with RA. In this study, CD8(+) T cells, mainly CD45RA(-) effector memory (EM) CD8(+) T cells, were increased significantly in the SF, but not in the peripheral blood from RA patients, compared with healthy controls. The synovial EM CD8(+) T cells were activated phenotypes with high levels of CD80, CD86, and PD-1, and had a proliferating signature in vivo upon Ki-67 staining, whereas the Fas-positive cells were prone to apoptosis. In addition, EM CD8(+) T cells in the SF were less cytotoxic, as they expressed less perforin and granzyme B. In particular, the proportions of synovial fluid mononuclear cells that were CCR4(+)CD8(+) T cells and IL-4-producing CD8(+) T cells (i.e., Tc2 cells) were significantly higher than those in peripheral blood mononuclear cells of patients with RA and healthy controls. In addition, the number of IL-10-producing CD8(+) suppressor T (Ts) cells increased significantly in the SF of RA patients. Especially, CD8(+) T cells were inversely correlated with disease activity. These findings strongly suggest that EM CD8(+) T cells in the SF are increased, likely because of inflammation, and they may be involved in modulating inflammation, thereby affecting the development and progression of RA.

High-throughput screening of saliva for early detection of oral cancer: a pilot study.

The success of tumour therapy depends considerably on early diagnosis. Therefore, we aimed to develop a widely available, cheap, non-invasive, high-throughput method suitable for screening high-risk populations, at least, for early signs of malignant transformation in the oral cavity. First, in order to identify suitable tumour marker candidates, we compared the protein patterns of five selected saliva samples obtained from healthy controls and tumour patients after electrophoretic separation, excised the bands that were consistently up-regulated in the tumour patients only, and performed matrix-assisted laser-desorption ionisation (MALDI)-time of flight (TOF) tandem mass spectrometry (MS/MS) analysis of the proteins in these bands after in-gel tryptic digestion. From the panel of proteins identified, we chose annexin 1 and peroxiredoxin 2 for further studies based on their presence in the saliva of all five oral cancer patients only. Then, we performed a homology search of protein databases using the primary sequence of each in silico tryptic fragment peptide of these two proteins as bait, and selected a unique peptide for each. Finally, we performed targeted MALDI-TOF MS peptide analysis in a blinded fashion on all samples obtained from 20 healthy controls and 22 tumour patients for the presence of these peptides. We found both peptides present in the saliva samples of all cancer patients only. Even though these tumour markers should be validated in a wider population, our results indicate that targeted MALDI-TOF MS analysis of unique peptides of putative saliva protein tumour biomarkers could be the method of choice for cost-efficient, high-throughput screening for the early detection of oral cancer.

Estudo fitoquímico e análise mutagênica das folhas e inflorescências de Erythrina mulungu (Mart. ex Benth. ) através do teste de micronúcleo em roedores / Phytochemical and mutagenic analysis of leaves and inflorescences of Erythrina mulungu (Mart. Ex Benth. ) through micronucleus test in rodents

Este trabalho teve como objetivo investigar a composição química, estabelecer a dose letal média (DL50) e avaliar os potenciais efeitos mutagênicos do extrato hidroalcoólico de folhas e inflorescências de Erythrina mulungu Mart. ex Benth por meio do teste de micronúcleo em medula óssea de camundongos. Os ensaios fitoquímicos foram realizados através de reações preliminares com mudança de coloração e/ou formação de precipitado; a DL50, por meio da administração intraperitoneal de três concentrações dos extratos, avaliando-se o número de óbitos após 48 horas e o teste de micronúcleo foi feito por meio do método do esfregaço, após exposição dos animais a cinco dias de tratamento. Os resultados fitoquímicos demonstraram presença de açúcares redutores, fenóis e taninos, proteínas e aminoácidos, flavonóides, alcalóides, depsídeos e depsidonas e derivados de cumarina em ambos os órgãos; saponinas espumídicas e esteróides e triterpenóides nas folhas e glicosídeos cardiotônicos e antraquinônicos e alcalóides nas inflorescências. Para a DL50 a folha demonstrou-se atóxica e a inflorescência moderadamente tóxica. Para o teste de micronúcleo, os resultados indicaram ausência de citotoxicidade e genotoxicidade dose-dependente para as folhas e independente da dose para as inflorescências. Assim, esses resultados sugerem que a planta, nas condições analisadas, possui potencial para induzir danos ao DNA.

This study aimed to investigate the chemical composition, to establish the mean lethal dose (LD50) and to assess the potential mutagenic effects of hydroalcoholic extract of leaves and inflorescences of Erythrina mulungu Mart. ex Benth by using micronucleus test in bone marrow of mice. Phytochemical assays were carried out through preliminary reactions with color change and/or precipitate formation; the LD50 was obtained by intraperitoneal administration of three concentrations of the extracts, assessing the number of deaths after 48 hours, and the micronucleus test was done by the smear method, after exposure of animals to five days of treatment. Phytochemical results showed the presence of reducing sugars, phenols and tannins, proteins and amino acids, flavonoids, alkaloids, depsides, depsidones and coumarin derivatives in both organs; foaming and steroidal saponins and triterpenes in the leaves and cardiotonic and anthraquinonic glycosides and alkaloids in the inflorescences. Considering the LD50, the leaf was atoxic and the inflorescence was moderately toxic. As regards the micronucleus test, results indicated absence of cytotoxicity while genotoxicity was dose-dependent for leaves and dose-independent for inflorescences. Thus, these results suggest that the plant, under the tested conditions, has the potential to induce damages to the DNA.