Comparison of Body Composition Methods for Estimating Body Fat Percentage in Lower Limb Prosthesis Users.

BACKGROUND: There is a dearth of literature evaluating the accuracy of Air Displacement Plethysmography (ADP) compared to Dual-energy X-ray Absorptiometry (DXA) for assessing body composition in individuals with lower limb amputations. Validity of ADP in persons with lower limb amputations must be established. OBJECTIVE: The objective of this study was to compare body composition in persons with lower limb amputations using the BOD POD® and DXA. METHODOLOGY: Body composition was performed on eleven lower limb prosthesis users (age 53.2±14.3 years, weight 81.9±22.3kg) using ADP and DXA with and without prosthesis. FINDINGS: Repeated measures ANOVA indicated no significant difference in body composition among and between trials, F(3,8)= 3.36, p= 0.075. There were no significant differences in Body Fat (BF) percentage with and without prostheses on the BOD POD (28.5±15.7% and 33.7±12.1%, respectively) nor the DXA (32.9±10.6% and 32.0±9.9%, respectively). Association between the BOD POD and DXA were greatest when prostheses were not worn compared to when they were worn. Bland-Altman plots indicate agreement between BOD POD® and DXA was greatest while wearing the prosthesis. CONCLUSION: This study is a first to compare total body fat percent between the BOD POD® and DXA in lower limb prosthesis users. BOD POD® report valid indices of BF%. Future work will utilize the BOD POD® in intervention studies for monitoring body composition changes across the continuum of rehabilitation.

Optimal application of compressive palatal stents following mesiodens removal in pediatric patients: A Randomized Controlled Trial.

BACKGROUND: There is no scientific evidence supporting the choice of a palatal stent in patients who underwent removal of an impacted supernumerary tooth. We aimed to investigate the effects of palatal stents in patients who underwent supernumerary tooth removal through a palatal approach and to suggest the optimal stent thickness and material. MATERIAL AND METHODS: We recruited 144 patients who underwent extraction of a supernumerary tooth between the maxillary anterior teeth. Subjects were assigned to a control group (CG) or one of four compressive palatal stent groups (CPSGs) classified by the thickness and material of the thermoplastic acrylic stent used. Palatal gingival swelling and objective indices (healing, oral hygiene, gingival, and plaque) were evaluated before surgery and on postoperative days (PODs) 3, 7, and 14; pain/discomfort and the Child Oral Health Impact Profile (COHIP) were assessed as subjective indices of the effects of the stent. RESULTS: The CPSGs showed faster healing than did the CG on PODs 7 (P<0.001) and 14 (P=0.043); swelling was measured by 1.64±0.88 mm and 4.52±0.39 mm, respectively. Although swelling was least in the 4-mm hard group (0.92±0.33 mm), the difference compared with that in the 2-mm hard group (1.01±0.18 mm) was not significant (P=0.077). The CPSGs showed better COHIP (P<0.001-0.036) and pain scores (P<0.001) than did the CG on PODs 1-3. CONCLUSIONS: Compressive palatal stents reduce discomfort by decreasing pain and alleviating swelling. Although a stent is effective regardless of its thickness and material, 2-mm hard stents maximized such positive effects with minimal discomfort.

Search for Sub-eV Sterile Neutrinos at RENO.

We report a search result for a light sterile neutrino oscillation with roughly 2200 live days of data in the RENO experiment. The search is performed by electron antineutrino (ν[over ¯]_{e}) disappearance taking place between six 2.8 GW_{th} reactors and two identical detectors located at 294 m (near) and 1383 m (far) from the center of the reactor array. A spectral comparison between near and far detectors can explore reactor ν[over ¯]_{e} oscillations to a light sterile neutrino. An observed spectral difference is found to be consistent with that of the three-flavor oscillation model. This yields limits on sin^{2}2θ_{14} in the 10^{-4}≲|Δm_{41}^{2}|≲0.5 eV^{2} region, free from reactor ν[over ¯]_{e} flux and spectrum uncertainties. The RENO result provides the most stringent limits on sterile neutrino mixing at |Δm_{41}^{2}|≲0.002 eV^{2} using the ν[over ¯]_{e} disappearance channel.

Thermal acclimation of rainbow trout myotomal muscle, can trout acclimate to a warming environment?

Climate change is a looming threat to the planet. Cold-water aquatic species will face significant physiological challenges due to elevated summer temperatures. Salmonids, such as rainbow trout (Oncorhynchus mykiss) maintain fidelity to native streams, limiting their ability to mitigate the impact of climate change through migration. We examined how rainbow trout swimming performance and muscle function were shaped by the thermal environment. We hypothesized that trout would show slower muscle contractile properties and slower swimming performance with long-term exposure to warmer water. For fish held at either 10 °C or 20 °C, maximum steady swimming speed (Ucrit) was determined, and contractile properties of both fast-twitch (white) and slow-twitch (red) myotomal muscle were examined. In addition, immunohistochemistry and quantitative PCR were used to assess changes in myosin content of the myotomal muscle in response to holding temperature. Rainbow trout exposed to warm water for six weeks displayed relatively limited thermal acclimation response. When tested at a common temperature (10 °C), 20 °C acclimated fish had modestly slower muscle performance compared to 10 °C acclimated fish. Significant differences in swimming performance and muscle contractile properties were primarily at colder test temperatures (e.g. 2 °C for muscle mechanics). Shifts in myosin heavy chain protein composition and myosin heavy chain gene expression in the swimming muscle were observed in white but not red muscle. Our results suggest that rainbow trout will have a limited ability to mitigate elevated environmental temperature through thermal acclimation of their myotomal or swimming muscle.

Fuel-Composition Dependent Reactor Antineutrino Yield at RENO.

We report a fuel-dependent reactor electron antineutrino (ν[over ¯]_{e}) yield using six 2.8 GW_{th} reactors in the Hanbit nuclear power plant complex, Yonggwang, Korea. The analysis uses 850 666 ν[over ¯]_{e} candidate events with a background fraction of 2.0% acquired through inverse beta decay (IBD) interactions in the near detector for 1807.9 live days from August 2011 to February 2018. Based on multiple fuel cycles, we observe a fuel ^{235}U dependent variation of measured IBD yields with a slope of (1.51±0.23)×10^{-43} cm^{2}/fission and measure a total average IBD yield of (5.84±0.13)×10^{-43} cm^{2}/fission. The hypothesis of no fuel-dependent IBD yield is ruled out at 6.6σ. The observed IBD yield variation over ^{235}U isotope fraction does not show significant deviation from the Huber-Mueller (HM) prediction at 1.3 σ. The measured fuel-dependent variation determines IBD yields of (6.15±0.19)×10^{-43} and (4.18±0.26)×10^{-43} cm^{2}/fission for two dominant fuel isotopes ^{235}U and ^{239}Pu, respectively. The measured IBD yield per ^{235}U fission shows the largest deficit relative to the HM prediction. Reevaluation of the ^{235}U IBD yield per fission may mostly solve the reactor antineutrino anomaly (RAA) while ^{239}Pu is not completely ruled out as a possible contributor to the anomaly. We also report a 2.9 σ correlation between the fractional change of the 5 MeV excess and the reactor fuel isotope fraction of ^{235}U.

Ultrasound-guided lung sliding sign to confirm optimal depth of tracheal tube insertion in young children.

BACKGROUND: The tip of the tracheal tube should lie at the mid-tracheal level after tracheal intubation in paediatric patients. Auscultation does not guarantee optimal positioning of the tracheal tube. We compared auscultation and the ultrasound-guided lung sliding sign to confirm optimal positioning of the tracheal tube in paediatric patients. METHODS: We studied 74 paediatric patients aged 0-24 months of ASA physical status 1-3 who were scheduled for elective surgery under general anaesthesia. All were randomly assigned to one of two groups: depth of tracheal tube confirmed by auscultation (Group A) or using the ultrasound-guided lung sliding sign (Group S). RESULTS: Optimal positioning of the tracheal tube was observed in 32 of 37 (87%) subjects in Group S and 24 of 37 (65%) subjects in Group A (difference in proportion, 22%; 95% confidence interval, 2-39%; P=0.030). Optimal depth correlated with patient height (adjusted coefficient=0.888, P<0.001). CONCLUSIONS: In paediatric patients younger than 24 months, use of the ultrasound-guided lung sliding sign was more accurate than auscultation for optimal positioning of the tracheal tube. CLINICAL TRIAL REGISTRATION: KCT 0003015.

Measurement of Reactor Antineutrino Oscillation Amplitude and Frequency at RENO.

The RENO experiment reports more precisely measured values of θ_{13} and |Δm_{ee}^{2}| using ∼2200 live days of data. The amplitude and frequency of reactor electron antineutrino (ν[over ¯]_{e}) oscillation are measured by comparing the prompt signal spectra obtained from two identical near and far detectors. In the period between August 2011 and February 2018, the far (near) detector observed 103 212 (850 666) ν[over ¯]_{e} candidate events with a background fraction of 4.8% (2.0%). A clear energy and baseline dependent disappearance of reactor ν[over ¯]_{e} is observed in the deficit of the measured number of ν[over ¯]_{e}. Based on the measured far-to-near ratio of prompt spectra, we obtain sin^{2}2θ_{13}=0.0896±0.0048(stat)±0.0047(syst) and |Δm_{ee}^{2}|=[2.68±0.12(stat)±0.07(syst)]×10^{-3} eV^{2}.

Extensive first-principles molecular dynamics study on Li encapsulation into C60 and its experimental confirmation.

The aim of increasing the production ratio of endohedral C60 by impinging foreign atoms against C60 is a crucial matter of the science and technology employed towards industrialization of these functional building block materials. Among these endohedral fullerenes, Li+@C60 exhibits a wide variety of physical and chemical phenomena and has the potential to be applicable in areas spanning the medical field to photovoltaics. However, currently, Li+@C60 can be experimentally produced with only ∼1% ratio using the plasma shower method with a 30 eV kinetic energy provided to the impinging Li+ ion. From extensive first-principles molecular dynamics simulations, it is found that the maximum production ratio of Li+@C60 per hit is increased to about 5.1% (5.3%) when a Li+ ion impinges vertically on a six-membered ring of C60 with 30 eV (40 eV) kinetic energy, although many C60 molecules are damaged during this collision. On the contrary, when it impinges vertically on a six-membered ring with 10 eV kinetic energy, the production ratio remains at 1.3%, but the C60 molecules are not damaged at all. On the other hand, when the C60 is randomly oriented, the production ratio reduces to about 3.7 ± 0.5%, 3.3 ± 0.5%, and 0.2 ± 0.03% for 30 eV, 40 eV, and 10 eV kinetic energy, respectively. Based on these observations we demonstrate the possibility of increasing the production ratio by fixing six-membered rings atop C60 using the Cu(111) substrate or UV light irradiation. In order to assess the ideal experimental production ratio, the 7Li solid NMR spectroscopy measurement is also performed for the multilayer randomly oriented C60 sample irradiated by Li+ using the plasma shower method combined with inductively coupled plasma atomic emission spectroscopy (ICP-AES). Time-of-flight mass spectroscopy measurements are also performed to cross check whether Li+@C60 molecules are produced in the sample. The resulting experimental estimate, 4% for 30 eV incident kinetic energy, fully agrees with our simulation results mentioned above, suggesting the consistency and accuracy of our simulations and experiments.

Evaluation of abundance of artificial radionuclides in food products in South Korea and sources.

Food samples are collected nationwide from January 2016 to February 2017 and their contents of artificial radionuclides are measured to address the growing concerns regarding the radioactive contamination of food products in Korea. Specifically, 900 food samples are collected for this study and their contents of representative artificial radionuclides 134Cs, 137Cs, 239,240Pu, and 90Sr are analyzed. The analysis shows that the activity concentrations of 137Cs in fish range from minimum detectable activity (MDA) to 340 mBq/kg of fresh weight. The concentration factor (CF) determined for 137Cs as a measure of its bioavailability is calculated to be ca. 74 and found to be very similar to that (100) recommended by the International Atomic Energy Agency. With an MDA of <0.221 mBq/kg, the results reveal that 239,240Pu values in fish are below the MDA. The activity concentrations of 137Cs and 90Sr are lower than the MDA in both shellfish and seaweed, while the activity concentrations of 239,240Pu in shellfish range from 0.26 to 2.18 mBq/kg, and for seaweed samples range from 2.07 to 3.38 mBq/kg. The atom ratios of 240Pu/239Pu in shellfish caught at the Korean coast vary from 0.209 to 0.237, with a mean of 0.227. The higher 240Pu/239Pu atom ratio determined in shellfish is thought to be caused by the plutonium transported from the Pacific Proving Grounds rather than other sources such as the Fukushima nuclear power plant accident. The activity concentrations of 137Cs in mushrooms are found to vary from 1.0 to 21.4 Bq/kg, with the highest concentrations observed in the Oak (shiitake) and Sarcodon asparatus. 134Cs is detected in three mushroom specimens collected from Jeju Island and about 3-3.6% of 137Cs present in the wild mushrooms native to the Jeju Island are introduced as a result of the Fukushima nuclear plant accident. The annual effective doses of 137Cs received through consumption of mushrooms and fish are 2.0 × 10-4 mSv yr-1 and 3.9 × 10-5 mSv yr-1, and those values are negligible compared to the annual effective doses limit of 1 mSv yr-1.

Treatment of M1a/M1b prostate cancer with or without radical prostatectomy at diagnosis.

BACKGROUND: The aim of the study was to evaluate survival and perioperative outcomes of metastatic prostate cancer (mPCa) patients treated with surgery or androgen deprivation treatment (ADT) only. METHODS: We retrospectively selected 47 metastatic PCa patients treated at a single center (Mayo Clinic, Rochester, MN) by two urologists (RJK and EK) between 2007 and 2014. Overall, 31 (66%) underwent radical prostatectomy (RP) with or without adjuvant therapies and 16 (34%) underwent ADT only. Surgical patients were treated by a single surgeon (RJK). Complications and functional outcomes were recorded for surgery group. Cancer-specific mortality (CSM) was analyzed by Kaplan-Meier estimation. Univariable Cox regression analyses were used to test the risk factors associated with CSM in mPCa patients treated with RP. RESULTS: Median age at diagnosis was 61 years. During median follow-up 38.8 months, 12 deaths were recorded. At 5 years, the overall CSM-free survival rate of the whole cohort was 57.9%. When patients were stratified according to the treatment, CSM-free survival rate at 5 years was 62% and 46% for patients who underwent surgery and ADT, respectively (P=0.3). Median length of stay was 3 days, with a 30 days readmission rate of 9.7%. The 30-day all complication rate was 29% (n=9). Specifically, we recorded: 2 lymphoceles (6.5%), 2 wound infection (6.5%), 2 ileus (6.5%), 2 hematoma (6.5%) and 1 anastomosis leak (3.2%). Within 90 days after surgery, 2 (6.5%) and 5 (16.1%) patients needed 1-2 supportive and 3 or more pads, respectively. However, continence was achieved by all treated patients during the follow-up period. CONCLUSIONS: We demonstrated the feasibility of local surgical treatment of primary tumor in mPCa patients. However, in the short term, no survival benefits have been observed for patients treated with surgery when compared with patients treated with ADT only. Further prospective studies are warranted to explore the treatment of M1a/M1b prostate cancer patients.

Impact of single-agent daily prednisone on outcomes in men with metastatic castration-resistant prostate cancer.

BACKGROUND: Despite palliative benefits and PSA responses, the objective clinical impact of daily oral prednisone (P) for metastatic castration-resistant prostate cancer (mCRPC) is unknown. We performed a pooled analysis of control arms of randomized trials that either did or did not administer single-agent P to evaluate its impact on overall survival (OS) and toxicities. METHODS: Individual patient data from control arms of randomized trials of men with mCRPC who received placebo or P+placebo post docetaxel were eligible for analysis. The impact of P on OS and severe toxicities was investigated in Cox regression models adjusted for known prognostic factors. Statistical significance was defined as P<0.05 and all tests were two sided. RESULTS: Data from the control arms of two randomized phase III trials were available totaling 794 men: the COU-AA-301 trial (n=394) administered P plus placebo and the CA184-043 trial (n=400) administered placebo alone. P plus placebo was not significantly associated with OS compared with placebo in a multivariable analysis (hazard ratio=0.89 (95% confidence interval 0.72-1.10), P=0.27). Other factors associated with poor OS were Eastern Cooperative Oncology Group (ECOG)-performance status (PS) ⩾1, Gleason score ⩾8, liver metastasis, high PSA, hypoalbuminemia and elevated lactate dehydrogenase (LDH). Grade ⩾3 therapy-related toxicities were significantly increased with P plus placebo compared with placebo (hazard ratio=1.48 (95% confidence interval 1.03-2.13), P=0.034). Other baseline factors significantly associated with a higher risk of grade ⩾3 toxicities were ECOG-PS ⩾1, hypoalbuminemia and elevated LDH. Fatigue, asthenia, anorexia and pain were not different based on P administration. CONCLUSIONS: P plus placebo was associated with higher grade ⩾3 toxicities but not extension of OS compared with placebo alone in men with mCRPC who received prior docetaxel. Except for the use of P with abiraterone to alleviate toxicities, the use of P should be questioned given its association with toxicities and resistance.

Positive Rates of Preliminary Crossmatches Among Transplantation Candidates Waitlisted for Different Organs in the Korean Network for Organ Sharing.

OBJECTIVE: For deceased-donor organ transplantations, negative T cell crossmatches (XMs) are mandatory for kidney and pancreas allocation in the Korean Network for Organ Sharing (KONOS) organ allocation system. Submission and periodic renewal of serum to the KONOS is required for all transplantation candidates of kidney or pancreas and these sera are distributed to 23 laboratories for preliminary XMs. We have investigated how sensitization status varies among transplantation candidates waitlisted for different organs. METHODS: Positive rates of T cell XMs performed during recent 1-year period (from March 2014 to February 2015) in the Korea Organ Donation Agency laboratory have been analyzed according to different organs. For 163 cases of deceased donors, 3605 recipients (22.1 recipients per one donor) were crossmatched using both of National Institutes of Health and antihuman globulin complement-dependent cytotoxicity (CDC) methods and flow cytometry method. RESULTS: T cell XM positive rates varied among transplantation candidates for different organs. The positive rate was high for kidney (485/3,145, 15.4%), and low for pancreas or kidney/pancreas (7/200, 3.5%), lung (0/41, 0%), and heart (11/221, 5.0%). Among XM-positive individuals, nearly two-thirds of the kidney transplantation candidates showed strong sensitization status with CDC+/flow+ results (64.5%), whereas pancreas or kidney/pancreas transplantation candidates more commonly showed weaker sensitization status with CDC-/flow+ results (85.7%). CONCLUSION: Kidney transplantation candidates show a much higher positive rate and stronger sensitization status than candidates for other organs. The results of this study would be useful for determining the number of candidates to be crossmatched for different organs.

cis-Prenyltransferase and Polymer Analysis from a Natural Rubber Perspective.

Dolichol and natural rubber are representative cis-polyisoprenoids in primary and secondary metabolism, respectively. Their biosynthesis is catalyzed by cis-prenyltransferase (CPT) by sequential condensations of isopentenyl diphosphates (IPPs) to a priming molecule. Although prokaryotic CPTs have been well characterized, the mechanism of eukaryotic CPTs in cis-polyisoprene biosynthesis was only recently revealed. It was shown that eukaryotes have evolved a unique protein complex, comprised of CPT and CPT-binding protein (CBP), to synthesize cis-polyisoprenoids. In the context of this new discovery, we found discrepancies in literature for CPT or CBP biochemical assays and in vivo CPT complementation using rer2 (yeast CPT) yeast mutant. Our study here shows that rer2 revertants occur at a frequency that cannot be disregarded and are likely accountable for the results that cannot be explained by the CPT/CBP heteroprotein complex model. To make a stable mutant, SRT1 gene (secondary CPT expressed at a basal level in yeast) was additionally deleted in the rer2Δ mutant background. This stable rer2Δ srt1Δ strain was then used to individually or simultaneously express Arabidopsis CPT1 (AtCPT1, At2g17570) and CBP (AtLEW1, At1G11755). We found that the simultaneous expression of Arabidopsis CPT1 and AtLEW1 effectively complements the rer2Δ srt1Δ strain, whereas the individual expression of AtCPT1 alone or AtLEW1 alone failed to rescue the yeast mutant. Microsomes from the dual expresser showed an efficient incorporation of IPPs into cis-polyisoprenoid (30% in 2h). These results showed that the CPT/CBP heteroprotein complex model is valid in Arabidopsis thaliana. Experimental details of these results are described in this methodology paper.

The metabolic response to a high-fat diet reveals obesity-prone and -resistant phenotypes in mice with distinct mRNA-seq transcriptome profiles.

OBJECTIVES: The aim of this study was to explore the phenotypic differences underpinning obesity susceptibility or resistance based on the metabolic and transcriptional profiling of C57BL/6J mice fed a high-fat diet (HFD). METHODS: The mice were fed either a normal diet or HFD for 12 weeks. After 6 weeks, the mice on HFD were classified as either obesity-prone (OP) or obesity-resistant (OR) depending on the body weight gain. RESULTS: Lipid profiles from plasma and liver significantly improved in OR mice relative to the OP group. Energy expenditure was greater in OR mice than in OP mice, with a simultaneous decrease in body fat mass. Epididymal white adipose tissue (eWAT) and liver were enlarged in OP mice (with visible immune-cell infiltration), but these effects were attenuated in OR mice compared with OP mice. Overall glucose metabolism was enhanced in OR mice compared with OP mice, including homeostasis model assessment for insulin resistance, plasma glucose and insulin concentrations, glucokinase activity and hepatic glycogen. Plasma adipokines and proinflammatory cytokines were upregulated in OP mice, and these changes were attenuated in OR mice. Transcriptomic profiles of eWAT and liver revealed common and divergent patterns of transcriptional changes in OP and OR mice, and pointed to differential metabolic phenotypes of OP and OR mice. There were substantial differences between OP and OR mice in molecular pathways, including atherosclerosis signaling, sperm motility, cAMP-mediated signaling in eWAT; and fibrosis, agranulocyte adhesion and diapedesis, and atherosclerosis signaling in liver. CONCLUSIONS: Taken altogether, the results provide robust evidence of major divergence in the transcriptomes, phenotypes and metabolic processes between obesity susceptibility and obesity resistance in the HFD-fed C57BL/6J mice.

Integrative systems analysis of diet-induced obesity identified a critical transition in the transcriptomes of the murine liver and epididymal white adipose tissue.

BACKGROUND: It is well known that high-fat diet (HFD) can cause immune system-related pathological alterations after a significant body weight gain. The mechanisms of the delayed pathological alterations during the development of diet-induced obesity (DIO) are not fully understood. METHODS: To elucidate the mechanisms underlying DIO development, we analyzed time-course microarray data obtained from a previous study. First, differentially expressed genes (DEGs) were identified at each time point by comparing the hepatic transcriptome of mice fed HFD with that of mice fed normal diet. Next, we clustered the union of DEGs and identified annotations related to each cluster. Finally, we constructed an 'integrated obesity-associated gene regulatory network (GRN) in murine liver'. We analyzed the epididymal white adipose tissue (eWAT) transcriptome usig the same procedure. RESULTS: Based on time-course microarray data, we found that the genes associated with immune responses were upregulated with an oscillating expression pattern between weeks 2 and 8, relatively downregulated between weeks 12 and 16, and eventually upregulated after week 20 in the liver of the mice fed HFD. The genes associated with immune responses were also upregulated at late stage, in the eWAT of the mice fed HFD. These results suggested that a critical transition occurred in the immune system-related transcriptomes of the liver and eWAT around week 16 of the DIO development, and this may be associated with the delayed pathological alterations. The GRN analysis suggested that Maff may be a key transcription factor for the immune system-related critical transition thatoccurred at week 16. We found that transcription factors associated with immune responses were centrally located in the integrated obesity-associated GRN in the liver. CONCLUSIONS: In this study, systems analysis identified regulatory network modules underlying the delayed immune system-related pathological changes during the development of DIO and could suggest possible therapeutic targets.

Pep19 drives epitope spreading in periodontitis and periodontitis-associated autoimmune diseases.

BACKGROUND AND OBJECTIVE: Epitope spreading is one of valid mechanisms operating in immunopathological processes of infection-induced autoimmune diseases. We hypothesized that the peptide 19 from Porphyromonas gingivalis heat shock protein (HSP) 60 (Pep19) may be the dominant epitope from which epitope-specific immune response to subdominant epitopes may diversify sequentially into autoimmune responses directed at human neoepitopes in P. gingivalis-induced periodontitis and autoimmune diseases. However, the exact feature and mechanism on how Pep19 may drive epitope spreading into human autoantigens in chronic periodontitis or P. gingivalis-induced experimental periodontitis has not been clarified. The present study was performed with the following specific aims: (i) to delineate retrospectively the features of epitope spreading by human cross-sectional analysis; (ii) to demonstrate prospectively the epitope spreading into new antigenic determinants in an ordered, predictable and sequential manner in experimental periodontitis; and (iii) to clarify the mechanism on how immunization with Pep19 may mobilize helper T cells or elicit B-cell responses to human autoantigens and neoantigen. MATERIAL AND METHODS: The study was devised for two independent investigations - a cross-sectional analysis on clinical subjects and a prospective analysis on experimental periodontitis - each being subdivided further into two additional independent observations. Cross-sectional dot immunoblot pattern against a panel of peptides of P. gingivalis HSP60 and human HSP60 was performed among age-dependent healthy subjects and between healthy subjects, patients with chronic periodontitis and patients with autoimmune disease, to identify epitope spreading. A peptide-specific T-cell line was established for phenotype analysis and for proliferation assay to an array of identical peptides. An identical prospective analysis was performed in P. gingivalis-induced experimental periodontitis or in Pep19-immunized mice. Cross-reactivity of anti-Pep19 monoclonal antibody was also investigated. RESULTS: A dominant immune response exclusively to Pep19 prevailed in healthy human subjects (before the age of 40) and mice that persisted in chronic periodontitis and autoimmune diseases without being replaced further by subsequent subdominant epitopes. A sequential epitope spreading provoked by Pep19 to subdominant autoantigen peptide 19 from human HSP60 (Hu19) in most healthy human subjects and mice, and to autoantigen peptide 9 from human HSP60 (Hu9) and neoantigen oxidized low-density lipoprotein (ox-LDL) in P. gingivalis-induced chronic periodontitis and autoimmune diseases could be demonstrated in a reproducible and predictable manner. T-cell proliferative activity to multiple autoantigens Hu19, Hu9 and ox-LDL, and cross-reactivity of anti-Pep19 monoclonal antibody to these epitopes may be proposed as cellular and molecular mechanisms responsible for the phenomenon. Moreover, the predictive value of Pep19 for Hu9 increased remarkably in the disease group when compared with that of the healthy group. CONCLUSION: Taken together, epitope spreading to Hu19, Hu9 and ox-LDL provoked by Pep19 could be proposed as a solid phenomenon observed in P. gingivalis-induced chronic periodontitis and infection-induced autoimmune diseases in a reproducible and predictable manner. T-cell proliferative activity to these peptides and cross-reactivity of anti-Pep19 antibodies to multiple human autoantigens could be proposed as cellular and molecular mechanisms responsible for this phenomenon.

Low Frequency of MKRN3 Mutations in Central Precocious Puberty Among Korean Girls.

Mutations of MKRN3, the gene encoding makorin RING-finger protein 3, lead to central precocious puberty (CPP). The aim of this study was to investigate mutations of the MKRN3 gene in Korean girls with CPP. Two hundred-sixty Korean girls with idiopathic CPP were included. Auxological and endocrine parameters were measured, and the entire MKRN3 gene was directly sequenced. MKRN3 gene analysis revealed one novel nonsense mutation (p.Gln281 *) and 6 missense variants (p.Ile100Phe, p.Gly196Val, p.Ile204Thr, p.Gln226Pro, p.Lys233Asn, and p.Ser396Arg). The novel nonsense mutation (p.Gln281 *) was a heterozygous C>T nucleotide change (c.841C>T) predicted to result in a truncated protein due to a premature stop codon in the MKRN3 gene. The nonsense mutation (p.Gln281 *) was only identified in one of the girls and her younger brother. Compared to previous reports on MKRN3 mutations in familial and sporadic cases of CPP, the present study reveals a relatively low number of MKRN 3 mutations in Korean girls with CPP. Larger samples of children with CPP and MKRN3 mutations are necessary in order to clarify whether the clinical course of puberty may differ as compared to idiopathic CPP.

The Association between Bone Age Advancement and Insulin Resistance in Prepubertal Obese Children.

OBJECTIVE: Obesity is associated with bone age (BA) advancement, but the underlying mechanisms remain unclear. The objective of this study was to investigate the association between BA maturation and insulin levels in obese children. METHODS: In this cross-sectional study of 93 prepubertal obese children, anthropometric data and hormonal values were measured. Subjects were divided into 2 groups based on the difference between BA and chronological age (CA) (noted as BA-CA). RESULTS: The study population included 39 (41.9%) males and 54 (58.1%) females with a mean age of 7.4±1.5 years. The advanced bone age group defined as BA-CA>1 year (n=44) had significantly higher HOMA-IR and fasting insulin levels, and lower quantitative insulin sensitivity check index (QUICKI). BA-CA was significantly correlated with fasting insulin (r=0.255, P=0.014), HOMA-IR (r=0.230, P=0.027), and QUICKI (r=- 0.301, P=0.003). Also, height SDS was significantly associated with ΔBA-CA (r=0.417, P<0.001). In the multiple regression analysis, HOMA-IR was identified as a significant independent predictor of BA-CA. CONCLUSION: Bone age is more advanced in obese children with hyperinsulinemia and insulin resistance. These findings suggest that insulin may affect skeletal maturation.

Effects of Coconut Materials on In vitro Ruminal Methanogenesis and Fermentation Characteristics.

The objective of this study was to evaluate the in vitro effects of coconut materials on ruminal methanogenesis and fermentation characteristics, in particular their effectiveness for mitigating ruminal methanogenesis. Fistulated Holstein cows were used as the donor of rumen fluid. Coconut materials were added to an in vitro fermentation incubated with rumen fluid-buffer mixture and timothy substrate for 24 h incubation. Total gas production, gas profiles, total volatile fatty acids (tVFAs) and the ruminal methanogens diversity were measured. Although gas profiles in added coconut oil and coconut powder were not significantly different, in vitro ruminal methane production was decreased with the level of reduction between 15% and 19% as compared to control, respectively. Coconut oil and coconut powder also inhibited gas production. The tVFAs concentration was increased by coconut materials, but was not affected significantly as compared to control. Acetate concentration was significantly lower (p<0.05), while propionate was significantly higher (p<0.05) by addition of the coconut materials than that of the control. The acetate:propionate ratio was significantly lowered with addition of coconut oil and coconut powder (p<0.05). The methanogens and ciliate-associated methanogens in all added coconut materials were shown to decrease as compared with control. This study showed that ciliate-associated methanogens diversity was reduced by more than 50% in both coconut oil and coconut powder treatments. In conclusion, these results indicate that coconut powder is a potential agent for decreasing in vitro ruminal methane production and as effective as coconut oil.