Subgingival microbiota of dogs with healthy gingiva or early periodontal disease from different geographical locations.

BACKGROUND: Periodontal disease is the most common oral disease of dogs worldwide and results from a complex interplay between plaque bacteria, the host and environmental factors. Recent studies have enhanced our understanding of the associations between the plaque microbiota and canine periodontal disease. These studies, however, were limited in their geographical reach. Thus associations between the canine oral microbiota and geographical location were investigated by determining the composition of subgingival plaque samples from 587 dogs residing in the United Kingdom (UK), United States of America (USA), China and Thailand using 454-pyrosequencing. RESULTS: After quality filtering 6,944,757 sequence reads were obtained and clustering of these at ≥98% sequence resulted in 280 operational taxonomic units (OTUs) following exclusion of rare OTUs (present at < 0.05% in all four countries). The subgingival plaque from dog populations located in the UK, USA, China and Thailand had a similar composition although the abundance of certain taxa varied significantly among geographical locations. Exploration of the effect of clinical status and age revealed a marked similarity among the bacteria associated with increased age and those associated with gingivitis: Young dogs and those with no gingivitis were dominated by taxa from the phyla Bacteroidetes and Proteobacteria whereas older dogs and those with moderate gingivitis were dominated by members of the Firmicutes. The plaque microbiota of small breed dogs was found to significantly differ to medium and large breeds and was dominated by species belonging to the Firmicutes. CONCLUSIONS: The bacterial associations with health, gingivitis and periodontitis were conserved across dogs from the UK, USA, China and Thailand. These bacterial signatures of periodontal health and disease have potential as biomarkers for disease detection.

Synthesis and biological evaluation in vitro and in mammalian cells of new heteroaryl carboxyamides as HIV-protease inhibitors.

New heteroaryl HIV-protease inhibitors bearing a carboxyamide spacer were synthesized in few steps and high yield, from commercially available homochiral epoxides. Different substitution patterns were introduced onto a given isopropanoyl-sulfonamide core modifying the type of heteroarene and the central core, with the presence of either H or benzyl group. Their in vitro inhibition activity against recombinant protease showed a general beneficial effect of carboxyamide moiety, the IC50 values ranging between 1 and 15nM. In particular benzofuryl derivatives showed IC50 values among the best for such structurally simple inhibitors. Docking analysis allowed to identify the favorable situation of such benzofuryl derivatives in terms of number of interactions in the active site, supporting the experimental results on activity. The inhibition activity of such molecules has been also evaluated in HEK293 cells expressing the protease fused to green fluorescent protein, by western blotting analysis, fluorescence microscopy and cytofluorimetry.

Flow cytometry and capillary electrophoresis analyses in ethanol-stressed Oenococcus oeni strains and changes assessment of membrane fatty acid composition.

AIMS: This study aimed to investigate the dynamics and physiological heterogeneity of Oenococcus oeni under different conditions, cell membrane fluidity and permeability variations, and assessment of changes in cell surface charging rates. METHODS AND RESULTS: Flow cytometry, membrane fatty acid analysis and capillary electrophoresis were performed to study ethanol-induced variations. Different physiological states were assessed, revealing cell subpopulations able to adapt and withstand to environmental stress, in order to recover their functionality. Moreover, total results demonstrated changes in cell surface and membrane fatty acid redistribution with a saturation degree and an unsaturated/saturated fatty acid ratio fairly steady in control and in different ethanol stresses. CONCLUSIONS: This study revealed a great variability among O. oeni strains and the importance to investigate the mechanisms by a multiparametric approach based on the structural and physiological bacterial adjustments in different stresses tolerance. SIGNIFICANCE AND IMPACT OF THE STUDY: Intermediate physiological state assessment in O. oeni with recovery possibility could be an important criterion for potential starter culture application. The flow cytometry application with changes in monitoring membrane fatty acid composition and in surface charging rates allowed the characterization of sorted subpopulations that may contribute to further understanding of diversity and heterogeneity in physiology of bacterial populations.

Stress response assessment of Lactobacillus sakei strains selected as potential autochthonous starter cultures by flow cytometry and nucleic acid double-staining analyses.

AIMS: The aim of this study was to apply the flow cytometry to Lactobacillus sakei strains, selected as potential autochthonous starters, to investigate dynamics and physiological heterogeneity of microbial behaviour under different stress conditions. METHODS AND RESULTS: A simultaneous nucleic acid double-staining assay was applied to discriminate cell populations in different physiological states after exposure to heat (50 and 55°C) and acid (pH 2·5 and 3·0) stresses. Alive cells with intact membranes, damaged cells still alive but with injured membranes, so with even a recovery ability, and dead cells with a permanent membrane damage were differentiated with a significant increase in damaged cells after stronger stress treatments. CONCLUSIONS: The existence and characteristics of subpopulations displaying heterogeneity in particular conditions are highly relevant, because specific subpopulations may show improved survival, changes and dynamics under stress conditions. SIGNIFICANCE AND IMPACT OF STUDY: This assay has potential for physiological research on lactic acid bacteria and for application in the food industry. The assessment of intermediate physiological states in Lb. sakei strains with recovery possibility could be an important criterion for application of potential starter cultures. Application of flow cytometry and characterization of sorted subpopulations may contribute to further understanding of diversity and heterogeneity in physiology of bacterial populations.

Continuous non-invasive measurement of stroke volume and cardiac index in infants and children: comparison of Impedance Cardiography NICaS® vs CardioQ® method.

BACKGROUND: Non-invasive measurement of cardiac output (CO) and cardiac index (CI) may become an important modality of moni- toring in pediatrics. Among the several methods proposed, impedance cardiography (ICG) has gained attention among the scientists. There are 2 basic technologies of ICG: thoracic body bioimpedance (TEB) and whole body electrical bioimpedance (WBEB). PURPOSE: The present study is aimed to test in infants and children the effectiveness of the Non-Invasive Cardiac System (NICaS®), a new ICG device working with a wrist-to-ankle configuration vs Car- dioQ® transesophageal doppler, a minimally invasive cardiac output monitor. METHODS: Whole-body bioimpedance measurements were obtained before and during the surgery with NICaS® and simultaneously with CardioQ®, demographic data were sampled, basement life monitoring were performed. RESULTS: Total of 42 patients aged from new born to 16 years old, were included in this study to evaluate heart rate (HR), stroke volume (SV), cardiac output (CO), cardiac index (CI), total peripheral resi- stance index (TPRI), total body water (TBW) and cardiac power index (CPI). 81 measurements were taken simultaneously by both devices from forty-two patients, with CardioQ® serving as the gold-standard for this evaluation and with NICaS®. The average values of CI in the study subjects for CardioQ® cardiac index (Q-CI) and NICaS® cardiac index (NI-CI) were 2.9±0.9 L/min/m² and 2.8±1.0 L/min/m2 respecti- vely (P<0.01). Overall, 2-tailed Pearson's correlation between NI-CI and Q-CI was r = 0.85. The Bland-Altman 1.96-standard deviation limit of agreement was -0.77 L/min and 0.87 L/min/m² with a small bias of 0.05 L/min/m². CONCLUSIONS: Good correlation was observed in pediatric patients for CI measured with NICaS® in comparison with CardioQ® device. Continuous non-invasive monitoring of NI-CI can be particularly in- teresting for the pediatric population.

A longitudinal assessment of periodontal health status in 53 Labrador retrievers.

OBJECTIVES: To determine the incidence and rates of progression of gingivitis and periodontitis in Labrador retrievers. MATERIALS AND METHODS: Fifty-three dogs, aged 1·1 to 5·9 years, had their periodontal health assessed every 6 months for up to 2 years. The extent of gingivitis and periodontitis was measured around the whole gingival margin of every tooth under general anaesthesia. RESULTS: All dogs had gingivitis at the initial assessment. The majority (64·2%) of tooth aspects had very mild gingivitis. The palatal/lingual aspect of all tooth types was most likely to show bleeding when probed: 63·0% of these aspects had mild or moderate gingivitis. Over 2 years, 56·6% of dogs developed periodontitis and dogs as young as 1·9 years were affected. There was a significant positive correlation between the proportion of teeth with periodontitis and age. In total, 124 teeth (5·7%) developed periodontitis; 88 (71·0%) of these were incisors. The palatal/lingual aspect of the incisors developed the disease first (2·8% of incisor aspects). CLINICAL SIGNIFICANCE: Periodontitis developed in regions that are difficult to see in conscious dogs implying that detection and treatment of disease requires periodic sedation or anaesthesia.

Antioxidant activities and quali-quantitative analysis of different Smallanthus sonchifolius [(Poepp. and Endl.) H. Robinson] landrace extracts.

Five landraces of Smallanthus sonchifolius [(Poepp. and Endl.) H. Robinson], known as yacon, were investigated in total phenolic content, antioxidant activity and chemical composition of ethanol extracts (EEs) and decoction extracts (DEs). The results demonstrated that DEs are rich in phenolic acids as caffeic acid, while the EEs show an higher amount of flavonoids, as luteolin 3',7-O-diglucoside and luteolin 7-O-glucoside. These flavonoid glycosides were identified for the first time in yacon extracts, together with apigenin and luteolin. The phytochemical profile explains the different antioxidant activities shown in our study. The landraces PER6-DE and PER4-DE showed the highest radical-scavenging activity and reducing power related to their polyphenolic contents. Results also show that yacon can be considered an important source of bioactive compounds with significant differences among the analysed landraces.

In vivo topical anti-inflammatory and in vitro antioxidant activities of two extracts of Thymus satureioides leaves.

Four extracts at increasing polarity were prepared from the leaves of Thymus satureioides Coss. (Labiatae) and assayed for the in vivo topical anti-inflammatory effect using the croton oil ear test in mice, and for in vitro both antioxidant (DPPH degrees test) and anti-bacterial (broth microdilution method) activities. The chloroform extract showed a topical anti-inflammatory activity (ID50=282 microg cm(-2)), only three times lower than that of the reference drug indomethacin (ID50=93 microg cm(-2)) and its active components were identified as ursolic and oleanolic acids. The methanol extract, showing a significant radical-scavenging effect (SC50=14.54 microg), was characterized by the isolation and identification of some flavonoids. On the contrary, the extracts did not show any anti-bacterial effect against four standard aerobial bacteria strains.

Left isomerism with tetralogy of Fallot and anomalous systemic and pulmonary venous connections.

The association of left isomerism with interruption and azygos continuation of the inferior caval vein, anomalous connection of the right pulmonary veins to the right atrium, an interatrial communication of sinus venosus type, and tetralogy of Fallot with right aortic arch and anomalous coronary artery has been observed in a 6-month-old patient. As far as we are aware, the association of these congenital cardiac malformations has not previously been reported in patients with isomeric left atrial appendages.