The effect of mobile personalised texting versus non-personalised texting on the caries risk of underprivileged adults: a randomised control trial.

BACKGROUND: In the Republic of Ireland (RoI), fluoridation has been effective and efficient for caries prevention at population level, regardless of income status; however, at individual level it still has limitations. This study aimed to compare personalised versus non-personalised text messaging on 'chance of avoiding new cavities' with the Cariogram, a computer-based caries risk assessment (CRA) model, in an economically disadvantaged adult population in the RoI. METHODS: The intervention was via a CRA summary letter plus 24 weekly personalised mobile-phone short text messages (text messages) based on the individual's CRA, compared with a non-personalised approach via a non-personalised letter and a predetermined, fixed set of 24 weekly text messages. The study was designed as a two-arm parallel-group, single-blinded (assessor), randomised controlled study in County Cork, RoI. The primary outcome was a comparison of 'chance of avoiding new cavities' calculated by the Cariogram with clinical examination, interview, CRT® (Ivoclar Vivadent, Liechtenstein) and three-day food diary between the two groups at follow-up. We combined stratified randomisation with blocked randomisation for 171 participants who completed baseline. Of them, 111 completed follow-up and were analysed (56 and 55 from the personalised and non-personalised groups, respectively). Due to protocol violations, both intent-to-treat (ITT) and per-protocol analyses were conducted. RESULTS: The ITT analysis did not show a personalised intervention effect on 'chance of avoiding new cavities'. Of the secondary outcome measures, only the stimulated saliva flow factor showed a personalised intervention effect, p = 0.036, OR = 0.3 (95% CI = 0.1, 0.9). The per-protocol analysis with 21 personalised and 33 non-personalised participants within two-message deviations showed no significant effect on 'chance of avoiding new cavities'. CONCLUSIONS: The null hypothesis in regard to the primary outcome for both ITT and per-protocol analyses was not rejected; however, as the minimal clinically important difference was included in the 95% CI for the per-protocol analysis, replication studies will be worth conducting to explore the potential of mobile devices for individual caries risk reduction. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry ( UMIN000027253 ) on 10 May 2017. The study was retrospectively registered.

Knowledge of caries risk factors/indicators among Japanese and Irish adult patients with different socio-economic profiles: a cross-sectional study.

BACKGROUND: A previous study has shown deficient knowledge of caries risk factors/indicators in a Japanese adult population regarded to have a high interest in preventive dentistry. No prior research has investigated caries risk knowledge in an Irish adult population. We hypothesise there may be unexpected differences or similarities in knowledge across countries with similar levels of economic development when comparing groups with different socio-economic and cultural profiles. Understanding what influences knowledge is important for the development of effective and efficient caries prevention strategies. The current paper aims to describe the knowledge of caries risk factors/indicators in two groups with different socio-economic profiles from two culturally distinct countries. METHODS: Cross-sectional surveys of adult dental patients were carried out in Japan and in the Republic of Ireland (RoI) using similar self-administered paper questionnaires. Patients were asked to identify caries risk factors/indicators from eight (Japan) or ten (RoI) listed items. The Japanese study involved 482 patients (aged ≥20 years) from 52 dental members of a nationwide web-based initiative Promoting Scientific Assessment in Prevention of Tooth Decay and Gum Disease (PSAP). The Irish study involved 159 patients (aged 20-69 years) accessing state-provided ('medical card') dental services from eight dental practices in County Cork. The two samples were compared. RESULTS: A higher proportion of Irish respondents identified 'Not visiting the dentist for check-up and cleaning' (OR 2.655; 99% CI 1.550, 4.547) and 'Not using fluoride' (OR 1.714; 99% CI 1.049, 2.802) than did Japanese respondents. A lower proportion of Irish respondents identified 'A reduced amount of saliva' (OR 0.262; 99% CI 0.159, 0.433) than Japanese respondents. Similarly shown in both studies were a persistent belief that 'Not brushing teeth properly' is a caries risk factor and a lack of knowledge on saliva buffering capacity as a caries risk factor. CONCLUSIONS: Deficiencies in knowledge which should be addressed: among the Japanese group, of dental check-up/cleaning visits and of fluoride use for caries prevention; among the Irish group, of saliva quantity as a caries risk factor. In addition, in both groups, we need to inform patients of the defensive role of saliva.

Identification of human ELOVL5 enhancer regions controlled by SREBP.

Fatty acid elongase 5 (ELOVL5) is an enzyme involved in the synthesis of polyunsaturated fatty acids. Sterol Regulatory Element-binding Protein (SREBP)-1 activates ELOVL5 and increases polyunsaturated fatty acid synthesis, which in turn negatively affects SREBP-1 expression. Thus, ELOVL5 has been established as an SREBP-1 target gene and an important component of the negative feedback loop of de novo lipogenesis. However, the human ELOVL5 promoter/enhancer has not been fully analyzed and the location of SREBP biding sites around the ELOVL5 gene has yet to be defined. Here we performed a detailed promoter/enhancer analysis of human ELOVL5 gene, and identified two new SREBP binding sites, one in the 10 kb upstream region and one in the exon 1. These two SRE motifs are conserved among mammals and the mechanism found in the present study by which SREBP activates ELOVL5 is considered to be common in mammals. Through these findings, we clarified the molecular mechanism how SREBP activates ELOVL5, an important regulator of de novo lipogenesis.

The critical role of neutral cholesterol ester hydrolase 1 in cholesterol removal from human macrophages.

RATIONALE: Hydrolysis of intracellular cholesterol ester (CE) is the key step in the reverse cholesterol transport in macrophage foam cells. We have recently shown that neutral cholesterol ester hydrolase (Nceh)1 and hormone-sensitive lipase (Lipe) are key regulators of this process in mouse macrophages. However, it remains unknown which enzyme is critical in human macrophages and atherosclerosis. OBJECTIVE: We aimed to identify the enzyme responsible for the CE hydrolysis in human macrophages and to determine its expression in human atherosclerosis. METHODS AND RESULTS: We compared the expression of NCEH1, LIPE, and cholesterol ester hydrolase (CES1) in human monocyte-derived macrophages (HMMs) and examined the effects of inhibition or overexpression of each enzyme in the cholesterol trafficking. The pattern of expression of NCEH1 was similar to that of neutral CE hydrolase activity during the differentiation of HMMs. Overexpression of human NCEH1 increased the hydrolysis of CE, thereby stimulating cholesterol mobilization from THP-1 macrophages. Knockdown of NCEH1 specifically reduced the neutral CE hydrolase activity. Pharmacological inhibition of NCEH1 also increased the cellular CE in HMMs. In contrast, LIPE was barely detectable in HMMs, and its inhibition did not decrease neutral CE hydrolase activity. Neither overexpression nor knockdown of CES1 affected the neutral CE hydrolase activity. NCEH1 was expressed in CD68-positive macrophage foam cells of human atherosclerotic lesions. CONCLUSIONS: NCEH1 is expressed in human atheromatous lesions, where it plays a critical role in the hydrolysis of CE in human macrophage foam cells, thereby contributing to the initial part of reverse cholesterol transport in human atherosclerosis.

Polyunsaturated fatty acids selectively suppress sterol regulatory element-binding protein-1 through proteolytic processing and autoloop regulatory circuit.

Sterol regulatory element-binding protein (SREBP)-1 is a key transcription factor for the regulation of lipogenic enzyme genes in the liver. Polyunsaturated fatty acids (PUFA) selectively suppress hepatic SREBP-1, but molecular mechanisms remain largely unknown. To gain insight into this regulation, we established in vivo reporter assays to assess the activities of Srebf1c transcription and proteolytic processing. Using these in vivo reporter assays, we showed that the primary mechanism for PUFA suppression of SREBP-1 is at the proteolytic processing level and that this suppression in turn decreases the mRNA transcription through lowering SREBP-1 binding to the SREBP-binding element on the promoter ("autoloop regulatory circuit"), although liver X receptor, an activator for Srebf1c transcription, is not involved in this regulation by PUFA. The mechanisms for PUFA suppression of SREBP-1 confirm that the autoloop regulation for transcription is crucial for the nutritional regulation of triglyceride synthesis.

Abrogation of neutral cholesterol ester hydrolytic activity causes adrenal enlargement.

We have previously demonstrated that neutral cholesterol ester hydrolase 1 (Nceh1) regulates foam cell formation and atherogenesis through the catalytic activity of cholesterol ester hydrolysis, and that Nceh1 and hormone-sensitive lipase (Lipe) are responsible for the majority of neutral cholesterol ester hydrolase activity in macrophages. There are several cholesterol ester-metabolizing tissues and cells other than macrophages, among which adrenocortical cells are also known to utilize the intracellular cholesterol for steroidogenesis. It has been believed that the mobilization of intracellular cholesterol ester in adrenal glands was facilitated solely by Lipe. We herein demonstrate that Nceh1 is also involved in cholesterol ester hydrolysis in adrenal glands. While Lipe deficiency remarkably reduced the neutral cholesterol ester hydrolase activity in adrenal glands as previously reported, additional inactivation of Nceh1 gene completely abrogated the activity. Adrenal glands were enlarged in proportion to the degree of reduced neutral cholesterol ester hydrolase activity, and the enlargement of adrenal glands and the accumulation of cholesterol esters were most pronounced in the Nceh1/Lipe double-deficient mice. Thus Nceh1 is involved in the adrenal cholesterol metabolism, and the cholesterol ester hydrolytic activity in adrenal glands is associated with the organ enlargement.

Targeting of neutral cholesterol ester hydrolase to the endoplasmic reticulum via its N-terminal sequence.

Neutral cholesterol ester hydrolase (NCEH) accounts for a large part of the nCEH activity in macrophage foam cells, a hallmark of atherosclerosis, but its subcellular localization and structure-function relationship are unknown. Here, we determined subcellular localization, glycosylation, and nCEH activity of a series of NCEH mutants expressed in macrophages. NCEH is a single-membrane-spanning type II membrane protein comprising three domains: N-terminal, catalytic, and lipid-binding domains. The N-terminal domain serves as a type II signal anchor sequence to recruit NCEH to the endoplasmic reticulum (ER) with its catalytic domain within the lumen. All of the putative N-linked glycosylation sites (Asn(270), Asn(367), and Asn(389)) of NCEH are glycosylated. Glycosylation at Asn(270), which is located closest to the catalytic serine motif, is important for the enzymatic activity. Cholesterol loading by incubation with acetyl-LDL does not change the ER localization of NCEH. In conclusion, NCEH is targeted to the ER of macrophages, where it hydrolyzes CE to deliver cholesterol for efflux out of the cells.

Hormone-sensitive lipase deficiency suppresses insulin secretion from pancreatic islets of Lep ob/ob mice.

It has long been a matter of debate whether the hormone-sensitive lipase (HSL)-mediated lipolysis in pancreatic beta-cells can affect insulin secretion through the alteration of lipotoxicity. We generated mice lacking both leptin and HSL Lep(ob/ob)/HSL(-/-) and explored the role of HSL in pancreatic beta-cells in the setting of obesity. Lep(ob/ob)/HSL(-/-) developed elevated blood glucose levels and reduced plasma insulin levels compared with Lep(ob/ob)/HSL(+/+) in a fed state, while the deficiency of HSL did not affect glucose homeostasis in Lep(+/+) background. The deficiency of HSL exacerbated the accumulation of triglycerides in Lep(ob/ob) islets, leading to reduced glucose-stimulated insulin secretion. The deficiency of HSL also diminished the islet mass in Lep(ob/ob) mice due to decreased cell proliferation. In conclusion, HSL affects insulin secretary capacity especially in the setting of obesity.

Identification of a novel member of the carboxylesterase family that hydrolyzes triacylglycerol: a potential role in adipocyte lipolysis.

Molecular mechanisms underlying lipolysis, as defined by mobilization of fatty acids from adipose tissue, are not fully understood. A database search for enzymes with alpha/beta hydrolase folds, the GXSXG motif for serine esterase and the His-Gly dipeptide motif, has provided a previously unannotated gene that is induced during 3T3-L1 adipocytic differentiation. Because of its remarkable structural resemblance to triacylglycerol hydrolase (TGH) with 70.4% identity, we have tentatively designated this enzyme as TGH-2 and the original TGH as TGH-1. TGH-2 is also similar to TGH-1 in terms of tissue distribution, subcellular localization, substrate specificity, and regulation. Both enzymes are predominantly expressed in liver, adipose tissue, and kidney. In adipocytes, they are localized in microsome and fatcake. Both enzymes hydrolyzed p-nitophenyl butyrate, triolein, and monoolein but not diolein, cholesteryl oleate, or phospholipids; hydrolysis of short-chain fatty acid ester was 30,000-fold more efficient than that of long-chain fatty acid triacylglycerol. Fasting increased the expression of both genes in white adipose tissue, whereas refeeding suppressed their expression. RNA silencing of TGH-2 reduced isoproterenol-stimulated glycerol release by 10% in 3T3-L1 adipocytes, while its overexpression increased the glycerol release by 20%. Thus, TGH-2 may make a contribution to adipocyte lipolysis during period of increased energy demand.

KLF15 Enables Rapid Switching between Lipogenesis and Gluconeogenesis during Fasting.

Hepatic lipogenesis is nutritionally regulated (i.e., downregulated during fasting and upregulated during the postprandial state) as an adaptation to the nutritional environment. While alterations in the expression level of the transcription factor SREBP-1c are known to be critical for nutritionally regulated lipogenesis, upstream mechanisms governing Srebf1 expression remain unclear. Here, we show that the fasting-induced transcription factor KLF15, a key regulator of gluconeogenesis, forms a complex with LXR/RXR, specifically on the Srebf1 promoter. This complex recruits the corepressor RIP140 instead of the coactivator SRC1, resulting in reduced Srebf1 and thus downstream lipogenic enzyme expression during the early and euglycemic period of fasting prior to hypoglycemia and PKA activation. Through this mechanism, KLF15 overexpression specifically ameliorates hypertriglyceridemia without affecting LXR-mediated cholesterol metabolism. These findings reveal a key molecular link between glucose and lipid metabolism and have therapeutic implications for the treatment of hyperlipidemia.

Caries experience of some countries and areas expressed by the Significant Caries Index.

OBJECTIVES: To calculate and present the caries prevalence for some countries/states among 12-year-olds, expressed as Significant Caries Index (SiC Index) and to analyse the relationship between the mean DMFT and the SiC Index for these countries. SiC Index is the mean DMFT of the one-third of a population with the highest caries values. METHODS: An Excel(R) application for calculating SiC was developed (http://www.whocollab.od.mah.se/expl/siccalculation.xls) and indices were calculated from the data collected for 14 countries and one state from the Country/Area Profile Programme (http://www.whocollab.od.mah.se/index.html). To investigate the provinces of a country that had already reached the proposed SiC Index goal of 3 DMFT among the 12-year-olds, data for 17 counties and a city from Sweden were collected and the respective mean DMFT and SiC Indices calculated. RESULTS: The mean DMFT varied from 1.0 to 8.5 and the SiC Index varied from 2.8 to 13.7 in the national data. Jamaica, Senegal and Sweden were the only three countries that showed SiC Indices that were less than 3 DMFT. The mean DMFT varied from 0.5 to 1.4 and the SiC Index varied from 1.4 to 3.6 in the Swedish county/city data examined. A strong linear relationship between the mean DMFT and the SiC Index was found for the populations presented in this study. CONCLUSION: The SiC Index is an indicator that reflects the situation among the most caries-exposed individuals and could be included in future population-based oral health surveys together with the mean DMFT.

Glycogen shortage during fasting triggers liver-brain-adipose neurocircuitry to facilitate fat utilization.

During fasting, animals maintain their energy balance by shifting their energy source from carbohydrates to triglycerides. However, the trigger for this switch has not yet been entirely elucidated. Here we show that a selective hepatic vagotomy slows the speed of fat consumption by attenuating sympathetic nerve-mediated lipolysis in adipose tissue. Hepatic glycogen pre-loading by the adenoviral overexpression of glycogen synthase or the transcription factor TFE3 abolished this liver-brain-adipose axis activation. Moreover, the blockade of glycogenolysis [corrected] through the knockdown of the glycogen phosphorylase gene and the resulting elevation in the glycogen content abolished the lipolytic signal from the liver, indicating that glycogen is the key to triggering this neurocircuitry. These results demonstrate that liver glycogen shortage activates a liver-brain-adipose neural axis that has an important role in switching the fuel source from glycogen to triglycerides under prolonged fasting conditions.

Apolipoprotein C-II deficiency with no rare variant in the APOC2 gene.

AIM: Familial apolipoprotein C-II (apoC-II) deficiency is a rare autosomal recessive disorder with marked hypertriglyceridemia resulting from impaired activation of lipoprotein lipase. In most cases of apoC-II deficiency, causative mutations have been found in the protein-coding region of APOC2; however, several atypical cases of apoC-II deficiency were reported to have markedly reduced, but detectable levels of plasma apoC-II protein (hereafter referred to as hypoapoC-II), which resulted from decreased promoter activity or improper splicing of apoC-II mRNA due to homozygous mutations in APOC2. Here we aim to dissect the molecular bases of a new case of hypoapoC-II. METHODS: We performed detailed biochemical/genetic analyses of our new case of hypoapoC-II, manifesting severe hypertriglyceridemia (plasma triglycerides, 3235 mg·dL(-1)) with markedly reduced levels of plasma apoC-II (0.6 mg·dL(-1)). RESULTS: We took advantage of a monocyte/macrophage culture system to prove that transcription of apoC-II mRNA was decreased in the patient's cells, which is compatible with the reported features of hypoapoC-II. Concomitantly, transcriptional activity of the minigene reporter construct of the patient's APOC2 gene was decreased; however, no rare variant was detected in the patient's APOC2 gene. Fifty single nucleotide variants were detected in the patient's APOC2, but all were common variants (allele frequencies ＞35%) that are supposedly not causative. CONCLUSIONS: A case of apoC-II deficiency was found that is phenotypically identical to hypoapoC-II but with no causative mutations in APOC2, implying that other genes regulate apoC-II levels. The clinical entity of hypoapoC-II is discussed.

Ablation of neutral cholesterol ester hydrolase 1 accelerates atherosclerosis.

Cholesterol ester (CE)-laden macrophage foam cells are the hallmark of atherosclerosis, and the hydrolysis of intracellular CE is one of the key steps in foam cell formation. Although hormone-sensitive lipase (LIPE) and cholesterol ester hydrolase (CEH), which is identical to carboxylsterase 1 (CES1, hCE1), were proposed to mediate the neutral CE hydrolase (nCEH) activity in macrophages, recent evidences have suggested the involvement of other enzymes. We have recently reported the identification of a candidate, neutral cholesterol ester hydrolase 1(Nceh1). Here we demonstrate that genetic ablation of Nceh1 promotes foam cell formation and the development of atherosclerosis in mice. We further demonstrate that Nceh1 and Lipe mediate a comparable degree of nCEH activity in macrophages and together account for most of the activity. Mice lacking both Nceh1 and Lipe aggravated atherosclerosis in an additive manner. Thus, Nceh1 is a promising target for the treatment of atherosclerosis.

Identification of neutral cholesterol ester hydrolase, a key enzyme removing cholesterol from macrophages.

Unstable lipid-rich plaques in atherosclerosis are characterized by the accumulation of macrophage foam cells loaded with cholesterol ester (CE). Although hormone-sensitive lipase and cholesteryl ester hydrolase (CEH) have been proposed to mediate the hydrolysis of CE in macrophages, circumstantial evidence suggests the presence of other enzymes with neutral cholesterol ester hydrolase (nCEH) activity. Here we show that the murine orthologue of KIAA1363, designated as neutral cholesterol ester hydrolase (NCEH), is a microsomal nCEH with high expression in murine and human macrophages. The effect of various concentrations of NaCl on its nCEH activity resembles that on endogenous nCEH activity of macrophages. RNA silencing of NCEH decreases nCEH activity at least by 50%; conversely, its overexpression inhibits the CE formation in macrophages. Immunohistochemistry reveals that NCEH is expressed in macrophage foam cells in atherosclerotic lesions. These data indicate that NCEH is responsible for a major part of nCEH activity in macrophages and may be a potential therapeutic target for the prevention of atherosclerosis.