A Novel Missense MEN1 Mutation in a Sporadic Case of Multiple Endocrine Neoplasia Type 1 Complicated with Papillary Thyroid Carcinoma.

Multiple endocrine neoplasia type 1 (MEN1) is a rare genetic disorder, resulting from MEN1 gene abnormalities, which causes tumors mainly in the endocrine glands. We experienced a sporadic case of MEN1 complicated with papillary thyroid carcinoma (PTC) and found a novel missense mutation in the patient's MEN1 gene. Her older sister, who showed no typical symptom of MEN1, had a history of PTC, suggesting the presence of another genetic factor involved in PTC development. This case suggests the importance of an individual's genetic background in the development of MEN1 complications.

AAA-ATPase valosin-containing protein binds the transcription factor SREBP1 and promotes its proteolytic activation by rhomboid protease RHBDL4.

Valosin-containing protein (VCP) is a member of AAA-ATPase superfamily involved in various cellular functions. To investigate the pathophysiological role of VCP in metabolic disorders, we generated knock-in mice bearing an A232E mutation in VCP, a known human VCP pathogenic variant. When heterozygous mutant mice (A232E/+) were fed a high-fat diet, we observed that fatty liver was ameliorated and the proteolytic processing of the transcription factor sterol regulatory element-binding protein 1 (SREBP1) was impaired. Further co-immunoprecipitation analysis in wildtype mice revealed interactions of VCP with SREBP1 and a rhomboid protease, RHBDL4, in the liver, and these interactions were attenuated in A232E/+ mice. Consistent with these results, we show that knockdown or chemical inhibition of VCP or RHBDL4 in human hepatocytes impaired the proteolytic processing of SREBP1. Finally, we found that knockdown of E3 ligases such as glycoprotein 78 and HMG-CoA reductase degradation protein 1 disrupted the interaction of VCP with SREBP1 and impaired the proteolytic processing of SREBP1. These results suggest that VCP recognizes ubiquitinylated SREBP1 and recruits it to RHBDL4 to promote its proteolytic processing. The present study reveals a novel proteolytic processing pathway of SREBP1 and may lead to development of new therapeutic strategies to treat fatty liver diseases.

Leptin sensitizing effect of 1,3-butanediol and its potential mechanism.

Leptin is an adipocyte-derived hormone that regulates appetite and energy expenditure via the hypothalamus. Since the majority of obese subjects are leptin resistant, leptin sensitizers, rather than leptin itself, are expected to be anti-obesity drugs. Endoplasmic reticulum (ER) stress in the hypothalamus plays a key role in the pathogenesis of leptin resistance. ATP-deficient cells are vulnerable to ER stress and ATP treatment protects cells against ER stress. Thus, we investigated the therapeutic effects of oral 1,3-butanediol (BD) administration, which increases plasma ß-hydroxybutyrate and hypothalamic ATP concentrations, in diet induced obese (DIO) mice with leptin resistance. BD treatment effectively decreased food intake and body weight in DIO mice. In contrast, BD treatment had no effect in leptin deficient ob/ob mice. Co-administration experiment demonstrated that BD treatment sensitizes leptin action in both DIO and ob/ob mice. We also demonstrated that BD treatment attenuates ER stress and leptin resistance at the hypothalamus level. This is the first report to confirm the leptin sensitizing effect of BD treatment in leptin resistant DIO mice. The present study provides collateral evidence suggesting that the effect of BD treatment is mediated by the elevation of hypothalamic ATP concentration. Ketone bodies and hypothalamic ATP are the potential target for the treatment of obesity and its complications.

Inhibitory effect of black cumin (Nigella sativa) seed essential oil on Fusobacterium nucleatum L-methionine-γ-lyase (L-methioninase) activity.

The enzyme L-methionine-γ-lyase is commonly found in a wide range of bacteria and catalyzes the α-elimination and γ-elimination of L-methionine to produce methyl mercaptan, α-ketobutyrate and ammonia. Black cumin seed essential oil (BC oil) reportedly exhibits deodorizing activity against methyl mercaptan. Therefore, we hypothesized that BC oil may also suppress methyl mercaptan production. In this study, we aimed to evaluate the inhibitory effect of BC oil on L-methionine-γ-lyase activity in Fusobacterium nucleatum. Recombinant L-methionine-γ-lyase was incubated under appropriate conditions with BC oil and its constituent thymoquinone. To analyze L-methionine-γ-lyase activity, α-ketobutyric acid and ammonia concentrations were determined. The concentrations of α-ketobutyric acid and ammonia were significantly decreased by 10 µg mL-1 of BC oil (P < 0.01) and 16.4 µg/mL of thymoquinone (P < 0.05). An enzyme kinetic assay showed a mixed inhibition pattern between L-methionine-γ-lyase and thymoquinone. In conclusion, BC oil not only had a deodorizing effect against methyl mercaptan but also an inhibitory effect on methyl mercaptan production through the suppression of L-methionine-γ-lyase activity. Thymoquinone may be mainly responsible for these effects of BC oil. Thus, application of natural BC oil may be adapted not only for medical use but also in other areas of life.

Effect of thymoquinone on Fusobacterium nucleatum­associated biofilm and inflammation.

Periodontitis affects oral tissues and induces systemic inflammation, which increases the risk of cardiovascular disease and metabolic syndrome. Subgingival plaque accumulation is a trigger of periodontitis. Fusobacterium nucleatum (FN) contributes to subgingival biofilm complexity by intercalating with early and late bacterial colonizers on tooth surfaces. In addition, inflammatory responses to FN are associated with the progression of periodontitis. Nigella sativa Lin. seed, which is known as black cumin (BC), has been used as a herbal medicine to treat ailments such as asthma and infectious diseases. The current study examined the inhibitory effect of BC oil and its active constituents, thymol (TM) and thymoquinone (TQ), on FN­associated biofilm and inflammation. FN­containing biofilms were prepared by co­cultivation with an early dental colonizer, Actinomyces naeslundii (AN). The stability and biomass of FN/AN dual species biofilms were significantly higher compared with FN alone. This effect was retained even with prefixed cells, indicating that FN/AN co­aggregation is mediated by physicochemical interactions with cell surface molecules. FN/AN biofilm formation was significantly inhibited by 0.1% TM or TQ. Confocal laser scanning microscopy indicated that treatment of preformed FN/AN biofilm with 0.01% of BC, TM or TQ significantly reduced biofilm thickness, and TQ demonstrated a cleansing effect equivalent to that of isopropyl methylphenol. TQ dose­dependently suppressed TNF­α production from a human monocytic cell line, THP­1 exposed to FN, yet showed no toxicity to THP­1 cells. These results indicated that oral hygiene care using TQ could reduce FN­associated biofilm and inflammation in gingival tissue.

Purification of a High Molecular Mass Protein in Streptococcus mutans.

Elucidation of a gene's function typically involves comparison of phenotypic traits of wild-type strains and strains in which the gene of interest has been disrupted. Loss of function following gene disruption is subsequently restored by exogenous addition of the product of the disrupted gene. This helps to determine the function of the gene. A method previously described involves generating a gtfC gene-disrupted Streptococcus mutans strain. Here, an undemanding method is described for purifying the gtfC gene product from the newly generated S. mutans strain following the gene disruption. It involves the addition of a polyhistidine-coding sequence at the 3' end of the gene of interest, which allows simple purification of the gene product using immobilized metal affinity chromatography. No enzymatic reactions other than PCR are required for the genetic modification in this method. The restoration of the gene product by exogenous addition after gene disruption is an efficient method for determining gene function, which may also be adapted to different species.

Localization of Sliding Movements Using Soft Tactile Sensing Systems with Three-axis Accelerometers.

This paper presents a soft tactile sensor system for the localization of sliding movements on a large contact surface using an accelerometer. The system consists of a silicone rubber base with a chamber covered by a thin silicone skin in which a three-axis accelerometer is embedded. By pressurizing the chamber, the skin inflates, changing its sensitivity to the sliding movement on the skin's surface. Based on the output responses of the accelerometer, the sensor system localizes the sliding motion. First, we present the idea, design, fabrication process, and the operation principle of our proposed sensor. Next, we created a numerical simulation model to investigate the dynamic changes of the accelerometer's posture under sliding actions. Finally, experiments were conducted with various sliding conditions. By confirming the numerical simulation, dynamic analysis, and experimental results, we determined that the sensor system can detect the sliding movements, including the sliding directions, velocity, and localization of an object. We also point out the role of pressurization in the sensing system's sensitivity under sliding movements, implying the ideal pressurization for it. We also discuss its limitations and applicability. This paper reflects our developed research in intelligent integration and soft morphological computation for soft tactile sensing systems.

Method for functional analysis of a gene of interest in Streptococcus mutans: gene disruption followed by purification of a polyhistidine-tagged gene product.

Typical methods for elucidating the function of a particular gene involve comparative phenotypic analysis of the wild-type strain and a strain in which the gene of interest has been disrupted. We previously described a simple method for the generation of a gene-disrupted strain in Streptococcus mutans by replacing the gene of interest with an antibiotic resistance marker gene. It is also crucial that the function lost following the gene disruption is restored by exogenous addition of the gene product, but purification of this product can be difficult and involve a complex series of steps. In this study, we describe a simple method for the purification of gene products following gene disruption in S. mutans. The method involves the expression of an additional polyhistidine tag at the C-terminus of the gene product. The target protein can be simply purified by immobilized metal affinity chromatography and applied to a restoration assay. This method utilizes the genomes of both the wild-type strain and the gene-disrupted strain as PCR templates to generate the DNA construct. Therefore, generation of the gene-disrupted strain is a prerequisite for the present procedure. The combination of gene disruption and gene product purification results in an efficient method for the analysis of gene function that could be further adapted to various other bacterial species.

Deodorizing Substance in Black Cumin (Nigella sativa L.) Seed Oil.

A deodorizing substance in black cumin (Nigella sativa L.), a spice for curry and vegetable foods in Southwest Asia, was examined. The essential oil prepared from the seeds of this plant exhibited strong deodorizing activity against methyl mercaptan, which is a main factor in oral malodor. After purification with silica gel column chromatography, the active substance in black cumin seed oil was identified as thymoquinone. This monoterpenic quinone functions as the main deodorizing substance in this oil against methyl mercaptan. Metabolite analysis suggested that the deodorizing activity may be generated by the addition of a reactive quinone molecule to methyl mercaptan. In the present study, the menthane-type quinone and phenol derivatives exhibited deodorizing activities via this mechanism.

Clinical efficacy and cost-effectiveness of lanthanum carbonate as second-line therapy in hemodialysis patients in Japan.

BACKGROUND AND OBJECTIVES: Lanthanum carbonate (LC) is a nonaluminum, noncalcium phosphate binder that is effective for hyperphosphatemia in dialysis patients. However, its efficacy and cost-effectiveness as second-line therapy have not been fully examined. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: We first conducted a multicenter, open-label, 16-week clinical trial to examine the effect of additive LC in 116 hemodialysis patients who had uncontrolled hyperphosphatemia with conventional phosphorus-lowering therapy alone. Based on these clinical data, a state transition model was developed to evaluate the benefits and costs associated with LC as second-line therapy. Reduced risks for cardiovascular morbidity and mortality among patients treated with LC arise through more of the population achieving the target phosphorus levels. Uncertainty was explored through sensitivity analysis. RESULTS: After 16 weeks of additive LC treatment, mean serum phosphorus levels decreased from 7.30 ± 0.90 to 5.71 ± 1.32 mg/dl, without significant changes in serum calcium or intact parathyroid hormone levels. A subsequent cost-effectiveness analysis showed that compared with conventional treatment, additive LC incurred an average additional lifetime cost of $22,054 per person and conferred an additional 0.632 quality-adjusted life years (QALYs). This resulted in an incremental cost-effectiveness ratio of $34,896 per QALY gained. Applying a cost-effectiveness threshold of $50,000 per QALY, a probabilistic sensitivity analysis showed that additive LC had a 97.4% probability of being cost-effective compared with conventional treatment. CONCLUSIONS: Our results indicate that the use of LC as second-line therapy would be cost-effective among hemodialysis patients with uncontrolled hyperphosphatemia in Japan.

Cinacalcet effectively reduces parathyroid hormone secretion and gland volume regardless of pretreatment gland size in patients with secondary hyperparathyroidism.

BACKGROUND AND OBJECTIVES: Cinacalcet is effective in reducing serum parathyroid hormone (PTH) in patients with secondary hyperparathyroidism. However, it has not been proven whether parathyroid gland size predicts response to therapy and whether cinacalcet is capable of inducing a reduction in parathyroid volume. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This 52-week, multicenter, open-label study enrolled hemodialysis patients with moderate to severe secondary hyperparathyroidism (intact PTH >300 pg/ml). Doses of cinacalcet were adjusted between 25 and 100 mg to achieve intact PTH <180 pg/ml. Ultrasonography was performed to measure the parathyroid gland size at baseline, week 26, and week 52. Findings were also compared with those of historical controls. RESULTS: Of the 81 subjects enrolled, 56 had parathyroid glands smaller than 500 mm(3) (group S) and 25 had at least one enlarged gland larger than 500 mm(3) (group L). Treatment with cinacalcet effectively decreased intact PTH by 55% from baseline in group S and by 58% in group L. A slightly greater proportion of patients in group S versus group L achieved an intact PTH <180 pg/ml (46 versus 32%) and a >30% reduction from baseline (88 versus 78%), but this was not statistically significant. Cinacalcet therapy also resulted in a significant reduction in parathyroid gland volume regardless of pretreatment size, which was in sharp contrast to historical controls (n = 87) where parathyroid gland volume progressively increased with traditional therapy alone. CONCLUSIONS: Cinacalcet effectively decreases serum PTH levels and concomitantly reduces parathyroid gland volume, even in patients with marked parathyroid hyperplasia.

Depressed expression of Klotho and FGF receptor 1 in hyperplastic parathyroid glands from uremic patients.

Fibroblast growth factor 23 (FGF23) exerts its effect by binding to its cognate FGF receptor 1 (FGFR1) in the presence of its co-receptor Klotho. Parathyroid glands express both FGFR1 and Klotho, and FGF23 decreases parathyroid hormone gene expression and hormone secretion directly. In uremic patients with secondary hyperparathyroidism (SHPT), however, parathyroid hormone secretion remains elevated despite extremely high FGF23 levels. To determine the mechanism of this resistance, we measured the expression of Klotho, FGFR1, and the proliferative marker Ki67 in 7 normal and 80 hyperplastic parathyroid glands from uremic patients by immunohistochemistry. All uremic patients had severe SHPT along with markedly high FGF23 levels. Quantitative real-time reverse transcription PCR showed that the mRNA levels for Klotho and FGFR1correlated significantly with their semi-quantitative immunohistochemical intensity. Compared with normal tissue, the immunohistochemical expression of Klotho and FGFR1 decreased, but Ki67 expression increased significantly in hyperplastic parathyroid glands, particularly in glands with nodular hyperplasia. These results suggest that the depressed expression of the Klotho-FGFR1 complex in hyperplastic glands underlies the pathogenesis of SHPT and its resistance to extremely high FGF23 levels in uremic patients.

Enhancement of oral moisture using tablets containing extract of Capparis masaikai Levl.

AIM OF THE STUDY: This study was performed to examine the effects of tablets containing an extract of Capparis masaikai Levl. (M-tablets) on enhancing oral moisture. SUBJECTS AND METHODS: The moistening effect of M-tablets was examined in 21 healthy subjects aged 25.1+/-2.4 (mean+/-S.D.) years in comparison with control tablets. After sucking tablets, the oral moisture was measured using a saliva wetness tester and a moisture checker. To evaluate the effects of the M-tablets on oral conditions, additional 50 subjects aged 30.6+/-7.5 years were examined. The subjects recorded changes in refreshment, oral moisture, ease in speaking, and taste of water using a visual analog scale (VAS). RESULTS: The L-SALIVO value of the M-tablet increased significantly from 1.83+/-0.17 (mean+/-S.E.M.) at baseline to 3.02+/-0.21 at 15 min (P<0.01). The Mucus((R)) value of the M-tablet also increased from 37.50+/-0.22 at baseline to 38.30+/-0.26 at 15 min (P<0.01). The VAS value for oral moisture increased significantly from 47.4+/-2.0 to 69.6+/-2.2 after taking the M-tablet (P<0.01). The VAS value for taste of water also increased from 50.0+/-1.1 to 66.7+/-3.2 (P<0.01). CONCLUSION: These results suggest that M-tablets are useful for enhancing oral moisture, which leads to improvement of oral conditions.

Carpal tunnel syndrome and plasma beta2-microglobulin concentration in hemodialysis patients.

As of the end of June 2005, 27 of 96 dialysis outpatients at our clinic had developed carpal tunnel syndrome (CTS). Of 19 patients who had undergone dialysis for 30 years or longer, 15 had CTS, whereas none of the 38 patients who had received dialysis for less than 10 years had CTS. These data reflect trends in CTS development: from 1983 the incidence of CTS increased for many years, but more recently there has been a decline in new cases of CTS. Comparison of the 27 CTS and 69 non-CTS dialysis patients at our clinic showed that those in the CTS group were older and had a longer duration of dialysis. Patients in the CTS group were found to have had a high plasma beta2-microglobulin (BMG) level in the distant past (15-21 years ago), but conversely had a much lower BMG level in recent years. Simple correlation analysis and multiple logistic regression analysis showed that the presence of CTS was correlated with high BMG levels in the distant past, in addition to age and duration of dialysis. These findings suggest that reduction of the plasma BMG level due to advances in dialysis therapy in recent years has contributed to the decreased incidence of CTS.

Cost-benefit analysis of a worksite oral-health promotion program.

This study was conducted to examine whether oral-health promotion programs provided as an occupational health service for employees were cost-beneficial for employers. The subjects were composed of 357 male workers (20-59 yr of age) who participated in oral-health promotion programs conducted at their workplaces between 1992 and 1997. The design of this study was a quasi-experimental study design in which the three programs (light: 1 visit; medium: 2-4 visits; and heavy: 5-6 visits) were compared through cost-benefit analysis conducted from the viewpoint of the employers. The programs consisted of oral-health checkups by dentists and oral-health education, including that on the proper brushing method, by dental hygienists. The costs of the program included direct costs for the payment of oral-health-care staff and for teaching materials, and indirect costs for the time for employee participation in the program (20 min/employee per visit). The accumulated dental expenses for the seven years were used to calculate benefits, which were determined, based on the differences between 0 visits and each program. The benefit/cost ratios of the three programs were -2.45, 1.46, and 0.73, respectively. These results suggest that a worksite oral-health promotion program of medium frequency is cost-beneficial for employers.

[2 dialysis cases which came to our clinic for the symptoms caused by hypercalcemia].

We'll report 2 dialysis cases which came to our clinic for the symptoms caused by hypercalcemia. Patients complained of sleeplessness, itching, headache, palpitation, apathy, akinesis, leanness, foot gangrene and so on. Hypercalcemia is one of the complication of vitamin D and calcium carbonate administration in chronic renal failure, though the frequency and risk are not clearly documented. Hypercalcemia aggravates the outcome of patients on dialysis and contributes to vascular calcification in long term. Recently various factors involving cardiovascular calcification are discussed, but first of all we must be very careful for the symptoms of hypercalcemia, and careful monitoring of plasma calcium concentration are recommended.

Usefulness of new wetness tester for diagnosis of dry mouth in disabled patients.

OBJECTIVES: The condition of dry mouth is an influential factor in the incidence of caries, periodontal disease, fungal infections, masticatory dysfunctions and denture function. Bedridden elderly and disabled persons often suffer from oral dryness and the aim of this study was to evaluate the usefulness of measuring the amount of moisture in the oral mucosa for clinical diagnosis of dry mouth in this group. SUBJECTS AND METHODS: The subjects were 20 elderly bedridden individuals, age range 65-89 years old, living in a nursing home and six healthy laboratory researchers, aged 20-46 years old, used as controls. Tongue dorsum moisture measurements were performed using a newly developed wetness tester (L-SALIVO), in which the wet portion was measured after 10 s. Further, clinical diagnosis of dry mouth was carried out using a clinical classification scale of the tongue mucosa (grade range, 0-3). RESULTS: It was possible to measure tongue dorsum moisture in all subjects with the wetness tester. The average moisture value was 0.1+/-0.2 mm in elderly subjects with a dry mouth grade of 2 (n = 8) or 3 (n = 12), while the average moisture value in the control subjects was 3.67+/-1.75 mm with a dry mouth grade of 0 (n = 4) or 1 (n = 2). Tester values and cliniical classification showed a positive co-relationship (r = 0.31, p < 0.05). CONCLUSIONS: Our results show that this new tester could be useful for evaluating oral dryness and diagnosing dry mouth.