Characterization of the salivary microbiome in healthy Thai children

Objective: To investigate the composition of the salivary microbiome of 50 healthy Thai children. Methods: A total 76 provinces in Thailand are grouped into 5 geographical clusters based on unique economics, foods and lifestyles. Geographical locations and the results of an oral assessment were also considered. Genomic DNA was extracted from stimulated saliva samples. Subsequently, amplicon libraries were prepared by 16 S Metagenomic Sequencing Library Preparation. The amplicons were sequenced using an Illumina Miseq platform followed by bioinformatics and statistical analyses.Results: The correlation between oral hygiene status and caries history varied from r^2=0.887 to r^2=0.999 in the geographical groups, suggesting oral hygiene status a strong association between caries history. Twenty taxonomic groups were found in all subjects and constituted 93.6%-96.5% of the microbiome. Of these, genus Veillonella and Prevotella showed significant differences in their proportions between the geographical groups(P<0.05). Furthermore, the proportion of Veillonella parvula, as well as Rothia aeria and Rothia dentocariosa tended to increase with worse oral hygiene status, which was also related to higher dental caries history.Conclusions: The differences in the salivary microbiome as related to geographic regions suggest that environmental factors, which may include dietary habits, could influence the predominant bacteria found in the mouth of Thai children, especially the genus Veillonella and Prevotella. The ratio of Veillonella parvula, Rothia aeria and Rothia dentocariosa may be indicators of worse oral hygiene status and future caries in this population.

Use of McGrath^(■) MAC Video Laryngoscope for Nasotracheal Intubation in Patients for Whom Intubation Was Expected to Be Difficult Due to the Limited Mouth Opening

McGrath? MAC video laryngoscope (McG) has been used for orotracheal intubation in both normal patients and patients for whom intubation was expected to be difficult, and has been reported to provide improved visibility of the glottis during tracheal intubation. There are some reports that normal nasotracheal intubation is easier with McG than with macintosh laryngoscope (ML). The usefulness of McG for nasotracheal intubation is beginning to be recognised. We experienced three cases using McG in patients for whom intubation was expected to be difficult due to the limited mouth opening and using McG for those patients enabled smooth nasotracheal intubation. McG provides good visual field during nasotracheal intubation, and is less invasive to the patient.

The Role of Thymidylate Synthase in Pemetrexed-Resistant Malignant Pleural Mesothelioma Cells

We established new pemetrexed-resistant cells originating from malignant pleural mesothelioma MSTO-211H cells to clarify the mechanism involved in pemetrexed resistance in malignant pleural mesothelioma. In the pemetrexed-resistant cells, only thymidylate synthase (TYMS) mRNA was overexpressed among other well-known molecular targets and chemosensitivity determinants of pemetrexed, and the role of the TYMS gene was ascertained by artificial regulation induced by specific siRNA. Silencing the TYMS expression partially restored the cytotoxicity of pemetrexed. The resistant cells did not display other gene alterations related to folate metabolism. We conclude that the primary mechanism imparting resistance to these cells is specific up-regulation of TYMS function. Further, the TYMS gene may serve as a useful biomarker for the prediction of pemetrexed chemosensitivity in patients with malignant pleural mesothelioma. We also investigated the efficacy of 1-(3-C-ethynyl-β-D-ribo-pento furanosyl)cytosine (ECyd) in overcoming pemetrexed resistance;this compound is presently undergoing clinical trials in the USA as TAS-106. ECyd had a similar antitumor effect on the resistant cells as that on the parental cells. In the clinical treatment of malignant pleural mesothelioma, ECyd promises to emerge as a novel drug.

Anesthetic Management for a Patient of Gorham’s Syndrome: The Vanishing Bone Disease

Gorham-Stout (GS) syndrome or the vanishing bone disease is a very rare chronic disease characterized by the destruction of the osseous matrix and proliferation of vascular structures. Review of the general anesthesia showed only a few cases till date. We report general anesthesia for tooth extraction in a 21-year-old male patient with Gorham-Stout syndrome. In this case, the most concerning issue was limited mouth opening due to mandible osteolysis and difficult intubation was anticipated. To anticipate difficult airway management, it is very important to consider the preoperative airway assessment including the cervical spine screening. In this case, the McGrath video laryngoscope prevented the anticipated difficult intubation due to the limited mouth opening due to mandible osteolysis.

Cell Proliferation Ability of Mouse Fibroblast-Like Cells and Osteoblast-Like Cells on a Ti-6Al-4V Alloy Film Produced by Selective Laser Melting

Successful regeneration of tissues and organs relies on the application of suitable substrates or scaffolds in scaffold-based regenerative medicine. In this study, Ti-6Al-4V alloy films (Ti alloy film) were produced using a three-dimensional printing technique called Selective Laser Melting (SLM), which is one of the metal additive manufacturing techniques. The thickness of produced Ti alloy film was approximately 250 μm. The laser-irradiated surface of Ti alloy film had a relatively smooth yet porous surface. The non-irradiated surface was also porous but also retained a lot of partially melted Ti-6Al-4V powder. Cell proliferation ability of mouse fibroblast-like cells (L929 cells) and mouse osteoblast-like cells (MC3T3-E1 cells) on both the surfaces of Ti alloy film was examined using WST assay. Both L929 and MC3T3-E1 cells underwent cell proliferation during the culture period. These results indicate that selective laser melting is suitable for producing a cell-compatible Ti-6Al-4V alloy film for biomaterials applications.

Molecular Basis of Neuroimmune Interaction in an In Vitro Coculture Approach

A team of researchers from Nagoya, Tokyo and Hamilton developed a unique technique for studying neuroimmune interaction with confocal laser scanning fluorescence microscopy several years ago. It relies on guiding immune and nerve cell interaction by creating an adhesive environment using an in vitro coculture dish. With their technique, they are able to study details of the mechanism of how nerve cells communicate with immune cells （mast cells and T lymphocytes） and vice versa. They showed that nerve-mast cell communication could occur in the absence of an intermediary transducing cell and that the neuropeptide substance P, operating v/a NK-1 receptors, was a soluble factor of this communication. In addition, recently, they showed that ATP which was released from activated mast cells mediated the activation of nerve cells. Further, with their technique, Nagoya＇s group was able to study details of the molecular mechanism of nerve-mast cell interaction. N-cadherin and CADM1 （cell adhesion molecule 1） appeared to mediate attachment and promoted the communication between mast cells and nerves predominantly. It would lead to new therapeutic modalities for diseases based on neuroimmune interaction such as neurogenic inflammation, intestinal bowel diseases, asthma, and autoimmune disorders. Cellular ＆ Molecular Immunology. 2008;5（4）：249-259.

Alanine Aminotransferase as the First Test Parameter for Wilson's Disease

Background and Aims: The liver is the first organ affected by toxic copper in the classical and severe hepatic forms of Wilson's disease(WD).Because their associated chronic liver damage is mostly asymptomatic,an intervention using a special test including serum alanine aminotransferase(ALT)activity is needed for detecting WD.Methods: Using the modified international criteria for the diagnosis of WD,45 patients were selected from the collective databases of our institutions,and 7 infants were reviewed from the literature.Two patients had the severe hepatic form,with nor-moceruloplasminemia and no mutations in ATP7B.The rapid ALT change during hemolytic anemia was adjusted for a baseline.The diagnostic potential of the ALT test was assessed from the age-dependent natural course of the liver damage of WD.Results: The natural course had three stages.ALTs were still low in some infants younger than 4 years-old.They were high in all children between the ages of 4 and 8 years-old;then,they reduced to low levels in some patients over 9 years of age.The high ALT stage represents chronic active hepatitis,and the sub-sequent low ALT stage is due to silent cirrhosis.The hepatic cop-per content is a reliable but invasive test,while urinary copper secretion is an alternative,non-invasive test for copper toxicosis of WD.The serum ceruloplasmin and ATP7B analyses are sub-type tests of WD.The response to anti-copper regimens is the final test result.Conclusions: ALTcould be the first parameter to test to detect WD in children between the ages of 4 and 8 years.

Phenotypes and Chronic Organ Damage May Be Different among Siblings with Wilson's Disease

Background and Aims:Cloning of ATP7B provided evidence that Wilson's disease is a hepatic copper toxicosis with a variety of extrahepatic complications.Affected siblings with the same genetic background and exposure to similar environmental factors may be a good model for the study of genotype-phenotype correlation.Methods:Twenty-three affected siblings in 11 families were selected from a database.The first phenotypes were determined according to the international proposal.The final types of chronic organ damage were re-evaluated for life-long management.Results:Phenotypes were identical in 5 of the families and different in 6 of the families.The acute hepatic phenotype H1 was found in 3 younger siblings and 1 older sibling.All survived an acute episode of hemolysis with underlying chronic liver disease.One also presented complication with neurological disease.The neurological phenotype N1 with neuropsychiatric symptoms and hepatic disease was found in 2 aged siblings of 1 family,in an older sibling in 3 families and in the oldest sibling in 1 family.Phenotypes in siblings were mainly split by either H1 occurring in random order or age-dependent N1.Types of chronic organ damage were identical in 8 of the families and different in 3 of the families.The same combination of chronic liver disease was found in 6 families and chronic liver disease complicated with neurological disease in 2 families.Split organ damage in siblings was found when an older sibling was complicated by neurological disease.There was no reverse combination of a younger sibling being complicated by neurological disease in any of the families.Conclusion:Phenotype combinations of siblings were mainly modified by externally-induced hemolytic episodes,while chronic organ damage in siblings was split by age-dependent neurological complications.

Copper Accumulates in Hemosiderins in Livers of Patients with Iron Overload Syndromes

In biology,redox reactions are essential and sometimes harmful,and therefore,iron metabolism is tightly regulated by cuproproteins.Since the state of copper in iron overload syndromes remains unclear,we investigated whether copper metabolism is altered in these syndromes.Eleven patients with iron overload syndromes participated in this study.The clinical diagnoses were aceruloplasminemia (n=2),hemochromatosis (n=5),ferroportin disease (n=2),and receiving excess intravenous iron supplementation (n=2).Liver specimens were analyzed using a light microscope and transmission electron microscope equipped with an X-ray analyzer.In addition to a large amount of iron associated with oxygen and phosphorus,the iron-rich hemosiderins of hepatocytes and Kupffer cells contained small amounts of copper and sulfur,regardless of disease etiology.Two-dimensional imaging clearly showed that cuproproteins were distributed homogenously with iron complexes within hemosiderins.Copper stasis was unlikely in noncirrhotic patients.The enhanced induction of cuproproteins by excess iron may contribute to copper accumulation in hemosiderins.In conclusion,we have demonstrated that copper accumulates in hemosiderins in iron overload conditions,perhaps due to alterations in copper metabolism.