Reply to: "The "pegylated" story continues – Perhaps because both ends (α2a and α2b) are true?"

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Occurrence of Bifidobacteriaceae in human hypochlorhydria stomach

open7noBackground: The human stomach, when healthy, is not a suitable host for microorganisms, but in pathological conditions such as gastritis, when gastric acid secretion is impaired, microbial overgrowth can be observed. Apart from Helicobacter pylori, the composition of microbiota, resident or exogenously introduced during neutral/high pH conditions, has not been investigated thoroughly. Thus, it is possible that Bifidobacteriaceae, important autochthonous and beneficial bacteria of human gastrointestinal microbiota, could over-colonize the stomach of hypochlorhydria patients suffering from autoimmune atrophic gastritis (AAG) or omeprazoletreated (OME) gastritis. This prompted us to characterize the Bifidobacteriaceae in such patients’ gastric microbiota and to study its abnormal colonization. Methods: Samples of gastric juices, and antrum and corpus mucosa from 23 hypochlorhydria patients (13 AAG and 10 OME) and from 10 control volunteers with base-line normochlorhydria, were cultivated in Brain Heart Infusion (BHI) and selective Bifidobacterium-Tryptone-Phytone-Yeast extract (Bif-TPY) media. The isolates were characterized by the fructose-6-phosphate phosphoketolase (F6PPK) test, electrophoresis of cellular proteins, the fermentation test, guanine-cytosine% DNA content, and DNADNA hybridization. Negative F6PPK isolates were characterized by order-specific polymerase chain reaction (PCR). Results: A total of 125 isolates, assigned to the Bifidobacteriaceae family on the basis of their morphology, were obtained from AAG and OME patients, but not from normal subjects. Of these isolates, 55 were assigned to the Bifidobacteriaceae family on the basis of their fructose-6-phosphoketolase (PPK) activity, PPK being the key taxonomic enzyme of this family. The remaining 70 isolates, which were PPK-negative, were attributed to the Actinomycetales order following specific primer PCR analysis. We observed a significantly higher abundance of Bifidobacteriaceae (Bifidobacterium dentium, Scardovia inopinata, and Parascardovia denticolens) in OME group than the AAG group. Furthermore, the Actinomycetales distribution was homogeneous for both hypochlorhydria patient groups. Conclusions: This study suggests that the Bifidobacteriaceae species, typically found in the oral cavity, readily colonizes the hypochlorhydria stomach of OME patients. The clinical relevance and the mechanism underlying this Bifidobacteriaceae presence in OME gastritis requires further functional studies.openPaola Mattarelli; Giovanni Brandi; Carlo Calabrese; Fabio Fornari; Gian Maria Prati; Bruno Biavati; Barbara SgorbatiPaola Mattarelli; Giovanni Brandi; Carlo Calabrese; Fabio Fornari; Gian Maria Prati; Bruno Biavati; Barbara Sgorbat

Gut microbiota profiles and characterization of cultivable fungal isolates in IBS patients

Studies so far conducted on irritable bowel syndrome (IBS) have been focused mainly on the role of gut bacterial dysbiosis in modulating the intestinal permeability, inflammation, and motility, with consequences on the quality of life. Limited evidences showed a potential involvement of gut fungal communities. Here, the gut bacterial and fungal microbiota of a cohort of IBS patients have been characterized and compared with that of healthy subjects (HS). The IBS microbial community structure differed significantly compared to HS. In particular, we observed an enrichment of bacterial taxa involved in gut inflammation, such as Enterobacteriaceae, Streptococcus, Fusobacteria, Gemella, and Rothia, as well as depletion of health-promoting bacterial genera, such as Roseburia and Faecalibacterium. Gut microbial profiles in IBS patients differed also in accordance with constipation. Sequence analysis of the gut mycobiota showed enrichment of Saccharomycetes in IBS. Culturomics analysis of fungal isolates from feces showed enrichment of Candida spp. displaying from IBS a clonal expansion and a distinct genotypic profiles and different phenotypical features when compared to HS of Candida albicans isolates. Alongside the well-characterized gut bacterial dysbiosis in IBS, this study shed light on a yet poorly explored fungal component of the intestinal ecosystem, the gut mycobiota. Our results showed a differential fungal community in IBS compared to HS, suggesting potential for new insights on the involvement of the gut mycobiota in IBS. KEY POINTS: Comparison of gut microbiota and mycobiota between IBS and healthy subjects Investigation of cultivable fungi in IBS and healthy subjects Candida albicans isolates result more virulent in IBS subjects compared to healthy subjects

Potent In Vitro Activity of Citrus aurantium Essential Oil and Vitis vinifera Hydrolate Against Gut Yeast Isolates from Irritable Bowel Syndrome Patients\u2014The Right Mix for Potential Therapeutic Use

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Archaeological and instrumental analyses of pottery and wood from the 'Leudo' wreck of Varazze, Italy

The wreck of a small boat found in the Ligurian sea (north west of Italy), known as 'Leudo del Mercante', and its cargo of pottery represent a unique case study of ceramic provenance and dating that can help in providing further insight about the coastal trade performed by minor ships in the Mediterranean regions. In this paper we review archaeological investigations on pottery and present results of instrumental analyses both on the cargo and on parts of the wooden hull. Our aim is to confirm the origin of the pottery and to try to solve the fundamental issue of dating. Elemental analysis of pottery was performed using PIXE (Particle Induced X-ray Emission); dating of wood samples from the hull was performed by radiocarbon Accelerator Mass Spectrometry (AMS). Both kinds of measurements were performed at the LABEC laboratory in Florence. The elemental composition of ceramic bodies shows that the pottery could have been produced in the area of Savona or Albisola, not far from the shipwreck site. With the detailed archaeological study of the cargo and using radiocarbon data, we can deduce that the boat was built, sailed and sunk most probably during the first half of the 16th century

PROROOT MINERAL TRIOXIDE AGGREGATE CEMENT USED AS A RETROGRADE FILLING WITHOUT ADDITION OF WATER: AN IN VITRO EVALUATION OF ITS MICROLEAKAGE

The aim of this in vitro study was to compare at different times the microleakage of roots filled with Proroot mineral trioxide aggregate (MTA) mixed with water as recommended by the manufacturer or filled only with powder without any previous hydration. Forty-four extracted single-rooted human teeth had their crown removed, and their roots were instrumented, filled with gutta-percha without cement, resected at 3 mm from the apex, and ultrasonically prepared with CK5 retrotips. The roots were randomly divided into 2 groups, group 1 (n 20) and group 2 (n 20) with 2 positive and 2 negative control roots. Teeth in group 1 were filled with white Proroot MTA prepared as suggested by the manufacturer, whereas in group 2, teeth were filled with white Proroot MTA by using only powder without any hydration. Microleakage of each root was measured at 24 hours and 1, 2, 4, and 12 weeks after filling with a fluid filtration device. No statistically significant difference (P .05) in microleakage between groups 1 and 2 was detected at any time except the 1-week period. The lack of water addition during the preparation of the cement in this study did not affect the in vitro sealing ability of Proroot MTA

Secondary Root Canal Treatment with Reciproc Blue and K-File: Radiographic and ESEM-EDX Analysis of Dentin and Root Canal Filling Remnants

Secondary root canal treatment requires the complete removal of filling materials with different chemical-physical properties. A newly developed single-use NiTi instrument (Reciproc Blue, RB) may be more effective in root canal retreatment. The aim of the present study was to evaluate morphology and composition of remnants after retreatment with RB compared to traditional K-File technique, in canals obturated with Thermafil/AH Plus. Twenty-four single-rooted human teeth were shaped with NiTi obturated with AH-Plus/Thermafil and retreated using RB NiTi instruments or manual K-Files. Radiographs were taken to evaluate endodontic space and radiopacity of residual filling-material before/after procedures. After retreatment, samples were longitudinally split and observed by environmental scanning electron microscopy connected to energy dispersive X-Ray spectroscopy (ESEM-EDX) to analyze the debris/remnant position, microchemistry, and dentinal surface morphology. Time for retreatments was recorded and compared using one-way ANOVA (p-value = 0.05). Radiopaque filling residuals were found in both groups. RB system resulted statistically faster than manual K-File in retreatment procedure (p < 0.001). Root canal space radiographic appearance obtained after retreatment with RB was wider than K-File (p < 0.05). ESEM-EDX revealed 4 different morphological dentin area. Area-1: debris-free with typical Ca, P, and N composition of dentin and detected in 70% of the surface. Area-2: presence of deproteinized smear layer free from N and debris in 15% of the surface. Area-3: a thick packed smear layer N-free and with fine debris consisting of trace elements from sealer in 10% of the surface. Area-4: packed with debris and trace elements. No difference was observed between both instruments regarding root canal space appearance and ESEM-EDX analysis. Both systems were able to remove filling material but created a dentine morphology composed of packed debris and filling materials embedded into the smear layer. Dentin surface composition resulted in collagen depleted by irrigation procedures. The reciprocating system required less time to complete retreatment

Metallurgical analysis and fatigue resistance of WaveOne and ProTaper Nickel–Titanium instruments

The aim of the study was to evaluate cyclic fatigue resistance of two NiTi instruments and to analyse their surface, fractographic and matrix morphology under ESEM/EDS and optical microscopy. WaveOne Primary and ProTaper Universal F2 brand new instruments were subjected to fatigue testing in an artificial canal with 5.0 mm radius and 60 angle of curvature. Seventy-two instruments were divided into three groups (n = 24), according to the selected kinematics: WaveOne using reciprocation (A); ProTaper using reciprocation (B) or rotation (C). Time to fracture was recorded. Data were analysed with ANOVA and Tukey test. ESEM/EDS analysis was conducted on new files to examine surface characteristics and on fractured fragments to identify the fractographic features. Metallographic analysis was performed with optical microscope on new instruments to evaluate alloy properties. Significant differences were found with Group A, which was statistically more resistant to cyclic fatigue (P\0.05) than the other groups. Surface analysis of new instruments showed both in WaveOne and ProTaper files the presence of deep milling marks. ESEM fractographic analysis of WaveOne showed multiple crack origins with an area of fatigue propagation wider than ProTaper instruments, in which a single crack origin could be detected. EDS analysis confirmed the equiatomic NiTi composition. Metallographic analysis under optical microscope revealed in WaveOne instruments the presence of nano-crystalline martensitic grains embedded in austenite matrix, presence which could not be found in ProTaper files. WaveOne NiTi files revealed higher resistance to fatigue stress, suggesting extended working time in clinical applications