Lavandula dentata L. essential oil: a promising antifungal and antibiofilm agent against oral Candida albicans.

Candida albicans is the main fungal species involved in oral candidiasis, and its increasing resistance to pharmacological treatment encourages the search for improved antifungal agents. Lavandula dentata L. essential oil (LD-EO) has been recognized for its antimicrobial activity, but little is known about its role against oral C. albicans. This study evaluated the antifungal and antibiofilm activities, mechanisms of action, and toxicity of LD-EO from Brazil against oral strains of C. albicans. Antifungal activity was assessed based on Minimum Inhibitory Concentration (MIC), Minimum Fungicidal Concentration (MFC), association study with miconazole (Checkerboard method), and sorbitol and ergosterol assays. Inhibition of biofilm formation and disruption of preformed biofilm were considered when studying the effects of the product. Additionally, the toxicity of LD-EO was evaluated by a hemolysis assay on human erythrocytes. Phytochemical analysis by gas chromatography-mass spectrometry identified eucalyptol (33.1%), camphor (18.3%), and fenchone (15.6%) as major constituents. The test substance showed mainly fungicidal activity (MIC100 = 8 µg/mL; MFC = 16 µg/mL), including against two miconazole-resistant isolates of C. albicans. The effects of LD-EO were synergistic with those of miconazole and appeared not to involve damage to the fungal cell wall or plasma membrane. Its effectiveness in inhibiting biofilm formation was higher than the effect of disrupting preformed biofilm. Finally, the product exhibited low hemolytic activity at MIC. Based on the favorable and novel results described here, LD-EO could constitute a promising therapeutic alternative for oral candidiasis, including miconazole-resistant cases.

Genome-wide identification and characterization of SNW/SKIP domain-containing proteins in plants.

Sessile organisms, such as plants, developed various ways to sense and respond to external and internal stimuli to maximize their fitness through evolutionary time. Transcripts and protein regulation are, among many, the main mechanisms that plants use to respond to environmental changes. SKIP protein is one such, presenting an SNKW interacting domain, which is highly conserved among eukaryotes, where SKI interacting protein acts in regulating key processes. In the present work, many bioinformatics tools, such as phylogenetic relationships, gene structure, physical-chemical properties, conserved motifs, prediction of regulatory cis-elements, chromosomal localization, and protein-protein interaction network, were used to better understand the genome-wide SNW/SKIP domain-containing proteins. In total, 28 proteins containing the SNW/SKIP domain were identified in different plant species, including plants of agronomic interest. Two main protein clusters were formed in phylogenetic analysis, and gene structure analysis revealed that, in general, the coding region had no introns. Also, expression of these genes is possibly induced by abiotic stress stimuli. Primary structure analysis of the proteins revealed the existence of an evolutionarily conserved functional unit. But physicochemical properties show that proteins containing the SNW/SKIP domain are commonly unstable under in vivo conditions. In addition, the protein network, demonstrated that SKIP homologues could act by modulating plant fitness through gene expression regulation at the transcriptional and post-transcriptional levels. This could be corroborated by the expression number of gene copies of SKIP proteins in many species, highlighting it's crucial role in plant development and tolerance through the course of evolution.

Evaluation of the antimicrobial activity of Eucalyptus radiata essential oil against Escherichia coli strains isolated from meat products.

The present study sought to evaluate the antimicrobial and anti-adherent potential of Eucalyptus radiata essential oil against food-borne strains of Escherichia coli. The study was performed using the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). In addition, the disk diffusion technique was used to evaluate the association of Eucalyptus radiata essential oil with synthetic antimicrobials. The Minimum Inhibitory Adherence Concentration (MIC) was also performed. The results revealed that E. radiata showed antimicrobial activity against the E. coli strains tested, with MIC values ranging from 500 µg/mL to 1000 µg/mL and MBC values ranging from 500 µg/mL to 1,024 µg/mL. As for the associations, it was observed that E. radiata oil exhibited a synergistic effect for some antibiotics, especially Ceftriaxone, with greater interference from the essential oil. Furthermore, it was effective in inhibiting the adherence of bacterial strains of E. coli, showing a more significant antibiofilm effect than the antibacterial agent 0.12% chlorhexidine digluconate. In summary, the essential oil of E. radiata showed antimicrobial potential against strains of E. coli of food origin, and can therefore, through in-depth studies, be used alone or in association with synthetic antimicrobials to combat infections caused by this pathogen.

Evaluation of the antibacterial activity of trans-anethole against Enterococcus cloacae and Enterococcus faecalis strains of food origin.

The present study sought to evaluate the antibacterial activity of trans-anethole against food-borne strains of Enterobacter cloacae and Enterococcus faecalis. The study was performed using Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC) methods, in addition, disc diffusion technique was used to evaluate the association of trans-anethole with synthetic antimicrobials. Minimum Inhibitory Concentration for Adherence (MICA) testing was also performed. The results revealed that trans-anethole presents no antibacterial activity at any of the concentrations used against the E. cloacae strains tested. However, trans-anethole presented antibacterial effect against five of the six E. faecalis bacterial strains tested, with MIC values ranging from 500 µg/mL to 1000 µg/mL. Further, when analyzing the MBC results against E. faecalis, it was observed that the compound presented values ranging from 500 µg/mL to 1000 µg/mL. As for the associations, it was observed that trans-anethole when combined with the antimicrobials ampicillin, gentamicin, ciprofloxacin, and ceftriaxone presented synergistic effect against most strains of E. faecalis. However, both trans-anethole and the control chlorhexidine (0.12%) presented no antibiofilm effects against strains of E. faecalis. In short, trans-anethole presented potential antibacterial against E. faecalis strains of food origin, and may upon further study, it may be used alone or in association with synthetic antimicrobials to combat infections caused by this bacterium.

Is there bacterial infection in the intact crowns of teeth with pulp necrosis of sickle cell anaemia patients? A case series study nested in a cohort.

AIM: To evaluate the presence of bacteria in permanent teeth with intact crowns (without caries, periodontal disease or dental trauma) in patients with sickle cell anaemia (HbSS genotype) by analysing their clinical, imaging and microbiological parameters. METHODOLOGY: This is a case series study nested in a cohort. In the first follow-up of this cohort study (Journal of Endodontics, 2013, 39, 177), 10 HbSS patients with at least one tooth with an intact crown and clinically diagnosed with pulp necrosis by pulse oximetry adapted for dentistry and a cold pulp sensitivity test (n = 27 teeth) were selected. Changes in the pulp chamber, root and periodontal ligament were identified in the tomographic analysis. Bacterial culture, staining for live and dead bacteria, and real-time polymerase chain reaction with 16S rRNA primers were used to identify the presence of bacteria. Culture sample collection was performed immediately after access to the pulp chamber. The microbiome was analysed with a MiSeq sequencer (Illumina, San Diego, CA). RESULTS: The diagnosis of pulp necrosis was confirmed clinically in 82% (22/27) of the teeth. The amount of bacterial load identified was less than 100 copies µL-1 in 23% (5/22) of the teeth with intact crowns and pulp necrosis. Thirteen bacterial species were identified that are commonly found in urinary tract infections, septicaemia and infective endocarditis. Only one of these species, Granulicatella adjacens, has also be found in primary endodontic infections. CONCLUSION: Prospective clinical, imaging and microbiological analyses suggest that pulp necrosis of teeth with intact crowns in HbSS patients is not associated with the presence of bacteria.

Flared Roots Reinforced With Bulk-fill Flowable Composite - Case Report.

This article presents a case report for the treatment of a patient with a flared root. The patient was treated with a bulk-fill flowable composite. This innovative approach seems to be efficient in reinforcing flared roots. The advantages and disadvantages of the technique are presented.

Anaemia, Serum Iron Concentrations and δ-Aminolevulinate Dehydratase Activity in Laying Hens Infected Naturally by Salmonella Gallinarum.

The aim of this study was to evaluate anaemia, serum iron concentrations and δ-aminolevulinate dehydratase (ALA-D) activity in laying hens infected naturally by Salmonella Gallinarum and having severe hepatic lesions. Liver and serum samples were collected from 27 laying hens (20 infected and seven uninfected). The δ-ALA-D activity, haematocrit and serum iron concentrations were evaluated. There were significant decreases in δ-ALA-D activity, haematocrit and serum iron concentrations (P <0.01) in birds infected by S. Gallinarum when compared with uninfected birds. There was a positive correlation (P <0.001) between serum iron concentration, haematocrit (r(2) = 0.82) and δ-ALA-D activity (r(2) = 0.75). A positive correlation was also observed between δ-ALA-D activity and haematocrit (r(2) = 0.78; P <0.01). Liver samples showed moderate focal coagulative necrosis associated with infiltration of lymphoplasmacytic cells, macrophages and heterophils. The anaemia in the infected hens may be related to reduction in δ-ALA-D activity and serum iron concentrations, since both are important for haemopoiesis.

Effects of eugenol on resting tension of rat atria

In cardiac and skeletal muscle, eugenol (μM range) blocks excitation-contraction coupling. In skeletal muscle, however, larger doses of eugenol (mM range) induce calcium release from the sarcoplasmic reticulum. The effects of eugenol are therefore dependent on its concentration. In this study, we evaluated the effects of eugenol on the contractility of isolated, quiescent atrial trabeculae from male Wistar rats (250-300 g; n=131) and measured atrial ATP content. Eugenol (1, 3, 5, 7, and 10 mM) increased resting tension in a dose-dependent manner. Ryanodine [100 µM; a specific ryanodine receptor (RyR) blocker] and procaine (30 mM; a nonspecific RyR blocker) did not block the increased resting tension induced by eugenol regardless of whether extracellular calcium was present. The myosin-specific inhibitor 2,3-butanedione monoxime (BDM), however, reversed the increase in resting tension induced by eugenol. In Triton-skinned atrial trabeculae, in which all membranes were solubilized, eugenol did not change resting tension, maximum force produced, or the force vs pCa relationship (pCa=-log [Ca2+]). Given that eugenol reduced ATP concentration, the increase in resting tension observed in this study may have resulted from cooperative activation of cardiac thin filaments by strongly attached cross-bridges (rigor state).

Effects of eugenol on resting tension of rat atria.

In cardiac and skeletal muscle, eugenol (µM range) blocks excitation-contraction coupling. In skeletal muscle, however, larger doses of eugenol (mM range) induce calcium release from the sarcoplasmic reticulum. The effects of eugenol are therefore dependent on its concentration. In this study, we evaluated the effects of eugenol on the contractility of isolated, quiescent atrial trabeculae from male Wistar rats (250-300 g; n=131) and measured atrial ATP content. Eugenol (1, 3, 5, 7, and 10 mM) increased resting tension in a dose-dependent manner. Ryanodine [100 µM; a specific ryanodine receptor (RyR) blocker] and procaine (30 mM; a nonspecific RyR blocker) did not block the increased resting tension induced by eugenol regardless of whether extracellular calcium was present. The myosin-specific inhibitor 2,3-butanedione monoxime (BDM), however, reversed the increase in resting tension induced by eugenol. In Triton-skinned atrial trabeculae, in which all membranes were solubilized, eugenol did not change resting tension, maximum force produced, or the force vs pCa relationship (pCa=-log [Ca2+]). Given that eugenol reduced ATP concentration, the increase in resting tension observed in this study may have resulted from cooperative activation of cardiac thin filaments by strongly attached cross-bridges (rigor state).

Characterization of ecto-ATPase activity in the surface of LLC-PK1 cells and its modulation by ischemic conditions.

BACKGROUND: The concentration of extracellular nucleotides is regulated by enzymes that have their catalytic site facing the extracellular space, the so-called ecto-enzymes. METHODS: We used LLC-PK1 cells, a well-characterized porcine renal proximal tubule cell line, to biochemically characterize ecto-ATPase activity in the luminal surface. The [γ-(32)P]Pi released after reaction was measured in aliquots of the supernatant by liquid scintillation. RESULTS: This activity was linear with time up to 20min of reaction and stimulated by divalent metals. The ecto-ATPase activity measured in the presence of 5mM MgCl(2) was (1) optimum at pH 8, (2) insensitive to different inhibitors of intracellular ATPases, (3) inhibited by 1mM suramin, an inhibitor of ecto-ATPases, (4) sensitive to high concentrations of sodium azide (NaN(3)) and (5) also able to hydrolyze ADP in the extracellular medium. The ATP:ADP hydrolysis ratio calculated was 4:1. The ecto-ADPase activity was also inhibited by suramin and NaN(3). The dose-response of ATP revealed a hyperbolic profile with maximal velocity of 25.2±1.2nmol Pixmg(-1)xmin(-1) and K(0.5) of 0.07±0.01mM. When cells were submitted to ischemia, the E-NTPDase activity was reduced with time, achieving 71% inhibition at 60min of ischemia. CONCLUSION: Our results suggest that the ecto-ATPase activity of LLC-PK1 cells has the characteristics of a type 3 E-NTPDase which is inhibited by ischemia. GENERAL SIGNIFICANCE: This could represent an important pathophysiologic mechanism that explains the increase in ATP concentration in the extracellular milieu in the proximal tubule during ischemia.

A case of severe pancreatitis complicated by Raoultella planticola infection.

A 45-year-old-male presented with severe pancreatitis. Two bacterial isolates obtained from peritoneal fluid and abdominal purulent secretion were identified to the species level by 15 biochemical tests and four supplementary tests as Raoultella planticola. Identification was confirmed by rpoB gene sequencing. R. planticola is difficult to identify in the clinical laboratory, and the clinical significance of this isolation remains uncharacterized. This is the first report of pancreatitis with a primary infection by R. planticola.

The role of HLA antigens in the development of paracoccidioidomycosis.

BACKGROUND: Paracoccidioidomycosis is a systemic granulomatous disease that involves primarily the lungs and may disseminate to other organs and systems. It is caused by Paracoccidioides brasiliensis, a fungus that exhibits reversible thermal dimorphism and whose natural habitat is presently unknown. There are two main clinical forms: the acute (subacute) juvenile form and the chronic adult form. The former runs a more rapid course and is more severe than the latter. This mycosis is found throughout Latin America. Brazil accounts for 80% of reported cases. Presumably P. brasiliensis thrives in humid and hot places, especially near forests or farms. The infection is endemic in certain areas, especially in Brazil, Colombia and Venezuela, where nearly 100% of the population show cutaneous paracoccidioidina positive skin tests, indicating previous contact with the fungus, although a small percentage show clinical manifestations of the disease. METHODS: We compared the expression of HLA class I antigens in a healthy group (control) and in a group of patients with paracoccidioidomycosis (chronic adult form) using the Terasaki lymphocytotoxicity test modified by Amos for HLA antigen analysis. AIMS: To discover indications of whether or not individual susceptibility to P. brasiliensis might depend on some specific immunological defect. RESULTS: There is no evidence of association between a specific HLA antigen and paracoccidioidomycosis in the subjects studied. Further investigations are recommended.

Dissolution evaluation of marketed tetracyclines solid oral dosage forms.

Tetracyclines are produced by several pharmaceutical industries, mainly as capsule dosage form. This paper aimed to evaluate quality performance of commercially available tetracycline and oxytetracycline hydrochloride capsules and doxycycline hydrochloride enteric-coated tablets by means of in vitro dissolution test. 36 samples from 12 different producers containing 500 mg of tetracycline and oxytetracycline and 100 mg of doxycycline were analyzed using the USP XXIII procedures. Thirty four samples (94.4%) were approved and the remaining two samples (5.6%) were rejected. Although only one of the producers had some samples rejected, this fact must be seriously taken into account, considering the need of a highly qualified industrial production of medicines.

Incidência de parasitoses em escolares da Escola Municipal de Educaçäo Infantil "Sant'Ana Itatiaia", Juiz de Fora-MG e sua possível correlaçäo com a qualidade da água para consumo / Parasitosis incidence in children from EMEI \"Sant'Ana Itatiaia\", Juiz de Fora-MG, Brazil and possible relationship with the presence of the parasites in source of water

SÄo apresentados os resultados de 23 exames parasitológicos de fezes coletadas com conservador MIF e analisadas pelo método preconizado por Hoffman, Pons & Janer no Laboratório de Saúde Pública/FFB/UFJF e, de 9 análises bacteriológicas da água consumida na residência de 7 escolares da EMEI "Sant'Ana Itatiaia" - Juiz de Fora - MG, segundo a metodologia empregada no Laboratório de Análise de Alimentos e Saneamento de Aguas/FFB/UFJF. O estudo foi desenvolvido nos meses de Agosto a Outubro de 1994 em escolares com idade entre 4 e 6 anos e de ambos os sexos. A percentagem das amostras de fezes positivas foi de 56,52 porcento e a de negativas de 43,48 porcento. A etiologia das enteroparasitoses de maior incidência na populaçäo estudada foi: Entamoeba coli (38,89 porcento), Ascaris lumbricoides (27,78 porcento), Trichuris trichiura (11,11 porcento), Giardia lamblia (11,11 porcento) e Entamoeba histolytica (11,11 porcento). Das amostras analisadas, 17,39 porcento revelaram a presença de mais de um parasita. Apesar de näo ter sido possível correlacionar a etiologia das enteroparasitoses com uma provável veiculaçäo hídrica das mesmas, verificou-se que 61,11 porcento das amostras de fezes examinadas revelaram a presença de protozoários e, ainda, que 60 porcento das amostras de água oriundas de outras fontes que näo o abastecimento público, foram consideradas impróprias para o consumo como bebida. Tais dados reforçam a necessidade de medidas administrativas eficientes para a implantaçäo e melhoria do saneamento básico, pois näo existe saúde sem infra-estrutura sanitária

Morphology of rat submandibular gland acinar secretory granules and their alteration during the acute phase of experimental Chagas' disease.

In a morphometric study at the electron microscope level immature submandibular glands of 8 normal male Holtzman rats were compared to those of 12 rats (46 days old) infected with the Y strain of Trypanosoma cruzi at day 18 of infection. In the fasted animals, the acinar cells of controls exhibited a variety of morphological types of immature secretory granules with lamellar substructure composed of swirling cords of electron-dense material, whereas in the infected rats mature secretory granules containing dense aggregates of short or long filamentous material were more abundant. These results suggest an acceleration of acinar maturation in infected animals.

Morphology of rat submandibular gland acinar secretory granules and their alteration during the acute phase of experimental Chagas' disease

In a morphometric study at the electron microscope level immature submandibular glands of 8 normal male Holtzman rats were compared to those of 12 rats (46 days old) infected with the Y strain of Trypanosoma cruzi at day 18 of infection. In the fasted animals, the acinar cells of controls exhibited a variety of morphological types of immature secretory granules with lamellar substructure composed of swirling cords of electron-dense material, whereas in the infected rats mature secretory granules containing dense aggregates of short or long filamentous material were more abundant. These results suggest and acceleration of acinar maturation in infected animals

Monoclonal antibody against a 52 K sperm surface protein inhibits sperm-zona pellucida interactions in the rat.

A major regulatory site for species specificity of fertilization in mammals lies at the level of sperm binding to the zona pellucida. This implies a high degree of complementarity between gamete and receptor molecules. These molecules support species-specific interactions between sperm and oocyte that lead to gamete fusion (fertilization). We identified a rat sperm head surface antigen using an IgG1 monoclonal antibody (HD1) against rat sperm of epididymis cauda. By electron microscopy the antigen was shown to be present on the plasma membrane surface of the sperm dorsal head. One- and two-dimensional immunoblotting analysis of sperm proteins demonstrated that HD1 reacted only with a 52 K molecule with a pI ranging from 6.6 to 7.2. The 52 K protein was first detected in situ by indirect immunofluorescence and showed to be underlining few elongated spermatids in testis. However, when the sperm reached the epididymis caput, the antigen was seen to be expressed on the dorsal surface of spermatozoa head. A similar fluorescence reaction was detected on sperms in the epididymis corpus and cauda. The specific spermatozoa-zona pellucida interaction was inhibited in the presence of monoclonal antibody HD1 in a sperm binding assay of in vitro fertilization.

Cystatin S in secretory granules fractions isolated from submandibular gland of infected rats by Trypanosoma cruzi.

We have measured the relative concentrations of cysteine proteinase in the granular fractions of submandibular glands of control and Trypanosoma cruzi infected rats by using a sensitive solid phase ELISA. Gland samples were homogenized in 0.34 M sucrose and 0.5 mM EDTA in 10 mM HEPES buffer at a pH of 7.4. The extract was centrifuged and filtered through Millipore filters to prepare a purer granular fraction. Immunochemical studies using antibody against cystatin S and electrophoretic analysis showed higher cystatin S levels in infected rats than in control. The role of this inhibitor during acute phase of Chagas disease is discussed.

Induction of synthesis of the rat cystatin S protein by the submandibular gland during the acute phase of experimental Chagas disease

Rats experimentally infected with Trypanosoma cruzi Y strain exhibited hypertrophy of the submandibular gland at 18 days after infection.SDS-PAGE of infected rats saliva revealed the presence of an additional band with an apparent molecular weight of about 13KDa. Electrophoresis of protein salivaand immunochemical analysis with antibody against rat cystatin S confirmed that the protein was identical to that induced by beta adrenergic stimulation

Induction of synthesis of the rat cystatin S protein by the submandibular gland during the acute phase of experimental Chagas disease.

Rats experimentally infected with Trypanosoma cruzi Y strain exhibited hypertrophy of the submandibular gland at 18 days after infection. SDS-PAGE of infected rats saliva revealed the presence of an additional band with an apparent molecular weight of about 13KDa. Electrophoresis of protein saliva and immunochemical analysis with antibody against rat cystatin S confirmed that the protein was identical to that induced by beta adrenergic stimulation.