A case of laparoscopic sigmoidectomy using thermography for colonic blood flow assessment.

BACKGROUND: Indocyanine green (ICG) fluorescence imaging is widely used in gastrointestinal surgery and is considered useful for reducing anastomotic leakage; however, because ICG remains in the tissue for a certain amount of time, we occasionally must re-evaluate colonic blood flow over a short time period during surgery. Herein, we verify the usefulness of thermography (TG) for evaluating colonic blood flow in a patient who underwent a laparoscopic sigmoidectomy for sigmoid colon cancer. CASE PRESENTATION: The patient is 43-year-old man who underwent laparoscopic resection of the sigmoid colon for colon cancer. After vascular treatment of the colonic mesentery, ICG/TG identified the boundary between ischemic and non-ischemic colon tissues. An additional 2 cm of colonic mesentery was resected because of the presence of a diverticulum noted at the intended site of oral anastomosis when attaching the anvil head. After additional vascular treatment of the colonic mesentery and administration of ICG, fluorescence was observed in the colon; however, TG identified the zone of the temperature transition on the surface of the colonic mesentery, even after additional colonic mesentery resection in the same region as previously observed. This zone was used as the cut-off line. There were no complications, such as anastomotic leakage, after the surgery. CONCLUSION: Although accumulation of similar cases is necessary, TG has the potential for use as an auxiliary diagnostic tool in clinical practice. TG can depict the presence or absence of blood flow based on surface temperature without the use of imaging agents, and is inexpensive and easy to perform.

Screening tests for predicting the prognosis of oral intake in elderly patients with acute pneumonia.

Many elderly patients with pneumonia have aspiration pneumonia. Therefore they must temporarily abstain from oral intake. However, it is difficult to predict whether or not they will be able to start oral intake. The reason is the standard method to evaluate deglutition about pneumonia patients has not been established. In this study we aimed to develop a simple and convenient method that predicts the prognosis of oral intake as nutrition among elderly patients with acute stage pneumonia. Participants were 77 inpatients fasting due to aspiration risk with acute pneumonia. (86.0 ± 7.7 years; range 68-105 years; men: n = 34, women: n = 43) during September 2011 and August 2013. Their consciousness levels were determined by Glasgow coma scale (GCS) and swallowing function and cough reflex were evaluated by repetitive saliva swallowing test (RSST), modified water swallow test, simple swallowing provocation test and cough test. Oral intake status at discharge was considered as the objective variable, and these tests were considered as explanatory variables. Then receiver operating characteristic (ROC) curve and the area under the curve (AUC) for each was done. From the ROC curve analysis, GCS ≥14 had the largest AUC (0.79) with a sensitivity and specificity of 0.71 and 0.80. That was followed by RSST ≥1, AUC (0.77) with a sensitivity and specificity of 0.81 and 0.67. These results suggest that GCS and RSST could be useful screening tests for prognostic prediction of oral intake capability in elderly patients with acute pneumonia.

Seasonal Changes in the Percentage of Rice Stripe Virus Viruliferous Laodelphax striatellus (Hemiptera: Delphacidae) in Paddy Fields in Japan.

Rice stripe disease, which is caused by Rice stripe virus (RSV), is one of the most serious viral diseases of rice. RSV is transmitted in a persistent manner by Laodelphax striatellus (Fallén). The incidence of the disease can be estimated from the density of viruliferous vectors. Understanding seasonal changes of the percentage of viruliferous L. striatellus can facilitate forecasting and controlling the disease. In paddies, the percentage of viruliferous insects fluctuated in phase with the rate of detection of RSV-infected rice; it gradually increased from July to August, plateaued or temporarily declined in September, and increased sharply on ratoons in October. These findings indicate that horizontal transmission of RSV from diseased plants to vector insects occurred frequently, and the insects acquired RSV from the ratoons. However, the percentages of viruliferous insects overwintering in poaceous weeds, the main hosts for L. striatellus in winter, were lower than those in ratoons. Few L. striatellus that acquired RSV from ratoons seemed to move to overwintering sites and transmit the virus to the next generation. However, there was a tendency for the percentages of viruliferous overwintering insects to be higher on paddy ridges than in river levees. Insects could probably move from ratoons to poaceous weeds when the weeds were near a paddy. Although increasing percentage of viruliferous insects on ratoons seem to have relatively little impact on RSV dynamics in the next crop season, appropriate weed management around paddies is still needed to reduce the incidence of rice stripe disease.

Immunoreactive antigens recognized in serum samples from mice intranasally immunized with Porphyromonas gingivalis outer membrane vesicles.

Outer membrane vesicles (OMVs) of the periodontopathic bacterium Porphyromonas gingivalis contain a wide range of virulence factors including lipopolysaccharide (LPS), fimbriae and gingipains. We have recently reported strong immunogenicity of OMVs using an intranasal vaccine mouse model. In the present study, we performed sub-immunoproteome analysis of OMV-immunized mouse serum samples from six different mice in order to identify immunodominant antigens. The combination of two-dimensional (2D) gel electrophoresis and mass spectrometry analysis identified OMV proteins of 53 spots on a 2D map, and it was notable that OMV proteins were largely distributed within a low pH range, in marked contrast to the ubiquitous distribution of outer membrane proteins. Western blot using the six serum samples after 2D electrophoresis revealed that all showed immunoreactivity to some diffuse signals at extremely low pH, which was similar to the distribution of immunoreactive signals when the A-LPS antibody was used. Mass spectrometry analysis also demonstrated that the signals corresponded to a wide range of virulence factors including A-LPS-modified proteins such as gingipains. Absorption of serum with LPS resulted in a dramatic reduction of immmunoreactivity. We conclude that LPS and A-LPS-modified proteins in OMVs carry immunodominant determinants and eventually elicit P. gingivalis-specific antibodies in mice.

Classification of sequential swallowing types using videoendoscopy with high reproducibility and reliability.

OBJECTIVE: This study aimed to classify sequential swallowing types using videoendoscopy (VE) avoiding radiation exposure and compare the results using videofluoroscopy (VF). DESIGN: Twenty-one healthy adults simultaneously underwent VF and VE during sequential straw drinking. Each discrete swallow was classified into an L-segmental type (laryngeal vestibule opens after swallow) or L-continuous type (laryngeal vestibule closure continues after swallow) using VF and a V-segmental type (velopharynx opens after swallow) or V-continuous type (velopharynx closure continues after swallow) using VE. Test-retest reproducibility and interrater and intrarater reliability were evaluated in ten healthy adults. RESULTS: Of 128 swallows, 94 were L-segmental and 34 were L-continuous types as per VF, whereas 95 were V-segmental and 33 were V-continuous types as per VE. The leading edge of the bolus at swallow onset was significantly deeper in L-continuous types (P = 0.001). Laryngeal vestibule closure on VF images corresponded to velopharyngeal closure on VE images for 127 of 128 swallows (κ = 0.98, P < 0.001). All subjects showed the same types of swallows in the first and the second studies. Both interrater and intrarater reliability were high. CONCLUSION: VE showed high reproducibility and reliability in the classification of sequential swallowing types.

Usefulness of a handheld nebulizer in cough test to screen for silent aspiration.

Cough test to screen for silent aspiration (SA) was reported, and the effectiveness was excellent. However, the device was rather large so that the portability was poor. So, the purpose of this study is to investigate the usefulness of a handheld nebulizer for the test and verify the reproducibility of the method. The subjects were 160 patients who were suspected of having dysphagia and underwent videofluorography (VF) or videoendoscopy (VE). They inhaled 1.0 % citric acid-physiologic saline orally for 1 min using a handheld nebulizer, and the examiner observed the number of coughs: more than five coughs was considered as negative (normal), while less than four coughs was regarded as positive. Among the subjects, 70 patients administered the cough test and VF or VE twice or more at some intervals. The k coefficient was calculated in reproducibility. Using the results of the VF or VE examination as the standards, for SA detection, the sensitivity was 0.86, specificity was 0.71, positive predictive value was 0.53, and negative predictive value was 0.93. The k coefficient was 0.79. In conclusion, the handheld nebulizer was useful in the cough test to screen for SA. Furthermore, satisfactory reproducibility was shown.

A missense mutation in the glucosamine-6-phosphate N-acetyltransferase-encoding gene causes temperature-dependent growth defects and ectopic lignin deposition in Arabidopsis.

To study the regulatory mechanisms underlying lignin biosynthesis, we isolated and characterized lignescens (lig), a previously undescribed temperature-sensitive mutant of Arabidopsis thaliana that exhibits ectopic lignin deposition and growth defects under high-temperature conditions. The lig mutation was identified as a single base transition in GNA1 encoding glucosamine-6-phosphate N-acetyltransferase (GNA), a critical enzyme of UDP-N-acetylglucosamine (UDP-GlcNAc) biosynthesis. lig harbors a glycine-to-serine substitution at residue 68 (G68S) of GNA1. Enzyme activity assays of the mutant protein (GNA1(G68S)) showed its thermolability relative to the wild-type protein. The lig mutant exposed to the restrictive temperature contained a significantly smaller amount of UDP-GlcNAc than did the wild type. The growth defects and ectopic lignification of lig were suppressed by the addition of UDP-GlcNAc. Since UDP-GlcNAc is an initial sugar donor of N-glycan synthesis and impaired N-glycan synthesis is known to induce the unfolded protein response (UPR), we examined possible relationships between N-glycan synthesis, UPR, and the lig phenotype. N-glycans were reduced and LUMINAL BINDING PROTEIN3, a typical UPR gene, was expressed in lig at the restrictive temperature. Furthermore, treatment with UPR-inducing reagents phenocopied the lig mutant. Our data collectively suggest that impairment of N-glycan synthesis due to a shortage of UDP-GlcNAc leads to ectopic lignin accumulation, mostly through the UPR.

Age-associated reduction of stimulatory effect of ghrelin on food intake in mice.

Aging is associated with a progressive decrease in appetite and food intake. We focused on the age-associated changes of the stimulatory effect of the appetite-regulating peptide, ghrelin. Food intake and the concentrations of acyl ghrelin and desacyl ghrelin in the plasma and in the stomach were measured with and without overnight fasting in young and old mice. Moreover, the food intake in response to the intraperitoneal administration of graded doses of acyl ghrelin was compared between young and old mice. Fasting drives food intake in young mice, but not in old mice. The concentrations of acyl ghrelin and desacyl ghrelin in the plasma and in the stomach were higher in the old mice than in the young mice. Food intake did not increase in old mice when stimulated by the administration of 1-3 nmol of acyl ghrelin, which could produce a significant increase in food intake in young mice. In conclusion, food intake did not increase in old mice after either overnight fasting or the administration of acyl ghrelin. The release and synthesis of ghrelin seem to be rather higher in old mice compared to young mice. These increases might be the results of compensation for the decline of receptor (and/or post-receptor) functions.

Research of masticatory function using hemiplegia simulator equipment.

BACKGROUND AND OBJECTIVE: Hemiplegic patients often exhibit a characteristic condition called Wernicke-Mann contracture. Therefore, the occlusal pattern in hemiplegic patients is considered to be adapted to stress because of this characteristic limb position. We created a sham Wernicke-Mann contracture in healthy individuals using hemiplegia simulator equipment and compared the functional occlusion in this position with that in the normal state to evaluate dynamic adaptive responses. METHODS: Wernicke-Mann contracture was simulated using a device to create sham hemiplegia (Manabi-tai, Hemiplegia Experiencing Set; Tokushu-iryo, Inc.). In addition to the measurement of the occlusal force using Dental Prescale(®) and Occluzer(®), the occlusion was evaluated using an electromyogram and stabilometer. RESULTS: There was a significant difference in the occlusal force between the normal state and during simulated hemiplegia. The surface electromyo-potential of the masseter muscle showed significantly higher values during simulated hemiplegia. It is significantly higher during simulated hemiplegia than in the normal state on the paralysed side, but not for the normal state on the non-paralysed side. The position and velocity vectors changed in the antero-posterior direction in the normal state but in the lateral direction during simulated hemiplegia. CONCLUSIONS: The hemiplegia simulator equipment is useful for research on hemiplegia, and that the occlusal balance is disturbed in the posture characteristic of hemiplegia.

Toward the realization of a better aged society: messages from gerontology and geriatrics.

1. BACKGROUND: Recent medical advancements, and improvements in hygiene and food supply have led to Japan having the longest life expectancy in the world. Over the past 50 years, the percentage of the elderly population has increased fourfold from 5.7% in 1960 to 23.1% in 2010. This change has occurred at the fastest rate in the world. Compared with France, where the percentage of the elderly population has increased just twofold in the past 100 years, Japanese society is aging at an unprecedented rate. In addition, the percentage of the very elderly (aged 75 years and over), comprising more frail people, exceeded 10% of the nation's population in 2008. In such a situation, many elderly Japanese wish to spend their later years healthy, and wish to achieve great accomplishments in their lives. To achieve that, rather than considering an aging population as a negative social phenomenon, we should create a society where elderly people can enjoy a healthy, prosperous life through social participation and contribution. Factors that hamper the elderly from leading a healthy life include various psychological and social problems occurring in older age, as well as a high incidence of diseases. Therefore, gerontology, which focuses on health promotion of the elderly by encompassing the study of social welfare, psychology, environment and social systems; and geriatrics, which focuses on health care of elderly people and carried out research, education and practices to promote health in the elderly, are becoming more important. Furthermore, along with a need for multidisciplinary care to support geriatric medicine, the development of a comprehensive education system for aged-care professionals is awaited. Thus, we should now recognize the importance of gerontology and geriatrics, and a reform of medical-care services should be made in order to cope with the coming aged society. Population aging is a global phenomenon. The actions being taken by Japan, the world's most aged society, have been closely watched by the rest of the world. Japan's aged society has been posing not only medical, nursing and welfare problems, but also complex problems closely associated with economy, industry and culture. Therefore, to solve these problems, a macroscopic integration and cooperation among industries, education institutions, administration and community through an interdisciplinary approach including medical science, nursing science, nursing care, study of social welfare, social science, engineering, psychology, economics, religion and ethics should be made. Regarding the promotion of gerontology, the "Committee for Establishing a Scientific Community for Sustainable Aged Society" of the Science Council of Japan also prepared a proposal and this was announced on 20 April 2011. 2. CURRENT SITUATION AND PROBLEMS: (1) Promotion of social participation and contribution of elderly people In Japan, the overall labor force rate is expected to decrease in the near future as a result of the low birth rate and high life expectancy. In contrast, many elderly people, particularly the young-old, have sufficient physical strength to fulfil their job duties and make a social contribution. For these people, a social structure where elderly people can work should be developed through re-educating the elderly and providing various job types. Promotion of social participation and contribution of the elderly is expected to cause a substantial increase in the labor force. Furthermore, it is also expected to contribute to not only the upturn of national economic activity through an increase in total consumption, but also a decrease in the number of elderly people who are likely to be in need of care. Therefore, in order for elderly people to be engaged in various social activities, strategies for developing a social structure for re-education, various employment statuses and employment opportunities should be prepared. However, as the total number of jobs is fixed, consideration should also be given to young workers. (2) Fostering medical specialists for aging Older people often suffer from many diseases, together with geriatric syndromes with multiple etiologies. Signs and symptoms vary according to each individual, and are often atypical; therefore, the patients visit different hospitals and receive many screening tests and prescriptions at the same time. To solve this problem, an effective screening system carried out by a primary-care doctor, and privacy-preserving medical data sharing among hospitals and clinics are needed. In a geriatric clinical setting, health-care professionals should be aware of the physical traits of older people who often develop not only dementia, but also geriatric syndromes, such as depression, falls and urinary incontinence, so that a holistic approach with consideration of nursing care is required. However, the existing Japanese medical education system is not prepared for medical professionals enabled to respond to the aforementioned requirements. Thus, the fostering of medical professionals who can provide comprehensive care - especially for the oldest-old - such as geriatric specialists and medical professionals who understand the principles of elderly care, is urgently needed. (3) Diagnosis of elderly-specific diseases and reform of medical-care services In Japan, the diagnostic system for elderly-specific diseases, including dementia, and reform of medical care services are markedly delayed. The current status concerning diagnosis, care and nursing should be investigated to collect academic data. In order to accumulate evidence for providing safe elderly care and nursing, the promotion of clinical research and a marked expansion of geriatric medical centers with high-level medical services are eagerly awaited. (4) Promotion of home-based care and multidisciplinary care To reduce the length of stay in acute hospitals, to reduce the physical burden of health-care professionals working at acute hospitals and to meet the demand of older people who prefer to remain in their own homes, further promotion of home-based care is needed. In addition, "multidisciplinary care" is increasingly needed to meet various demands in the medical care and welfare of the elderly. It is considered important to share countermeasures against the problems of disease prevention, medicine, care and welfare among health-care professionals in medicine, care and welfare, and cooperate by making the best use of health-care professionals' specialties. 3. CONTENTS OF THE PROPOSAL: The subcommittee for aging, thus, provided the following proposal: 1 Development and promotion of systems that enable elderly people to participate socially and make a contribution using an interdisciplinary approach among the various areas, including nursing science, nursing care, study of social welfare, social science, psychology, economics, religion and ethics, as well as medical sciences; 2 Promotion of gerontology, reform and enhancement of geriatrics in undergraduate, postgraduate and lifelong education; 3 Building geriatric medical centers in each area, and accumulating large-scale evidence of geriatric diseases and geriatrics; and 4 Structural development and promotion of home-based care and multidisciplinary care. Through implementation of the above measures, Japan is expected to function as a successful example for the rest of the world.

Essential role of caspase-8 in p53/p73-dependent apoptosis induced by etoposide in head and neck carcinoma cells.

BACKGROUND: Caspase-8 is a key upstream mediator in death receptor-mediated apoptosis and also participates in mitochondria-mediated apoptosis via cleavage of proapoptotic Bid. However, the role of caspase-8 in p53- and p73-dependent apoptosis induced by genotoxic drugs remains unclear. We recently reported that the reconstitution of procaspase-8 is sufficient for sensitizing cisplatin- but not etoposide-induced apoptosis, in chemoresistant and caspase-8 deficient HOC313 head and neck squamous cell carcinoma (HNSCC) cells. RESULTS: We show that p53/p73-dependent caspase-8 activation is required for sensitizing etoposide-induced apoptosis by utilizing HOC313 cells carrying a temperature-sensitive p53G285K mutant. Restoration of wild-type p53 function under the permissive conditions, together with etoposide treatment, led to substantial transcriptional activation of proapoptotic Noxa and PUMA, but failed to induce apoptosis. In addition to p53 restoration, caspase-8 reconstitution was needed for sensitization to etoposide-induced apoptosis, mitochondria depolarization, and cleavage of the procaspases-3, and -9. In etoposide-sensitive Ca9-22 cells carrying a temperature-insensitive mutant p53, siRNA-based p73 knockdown blocked etoposide-induced apoptosis and procaspase-8 cleavage. However, induction of p73 protein and up-regulation of Noxa and PUMA, although observed in Ca9-22 cells, were hardly detected in etoposide-treated HOC313 cells under non-permissive conditions, suggesting a contribution of p73 reduction to etoposide resistance in HOC313 cells. Finally, the caspase-9 inhibitor Ac-LEHD-CHO or caspase-9 siRNA blocked etoposide-induced caspase-8 activation, Bid cleavage, and apoptosis in both cell lines, indicating that p53/p73-dependent caspase-8 activation lies downstream of mitochondria. CONCLUSIONS: we conclude that p53 and p73 can act as upstream regulators of caspase-8, and that caspase-8 is an essential mediator of the p53/p73-dependent apoptosis induced by etoposide in HNSCC cells. Our data suggest the importance of caspase-8-mediated positive feedback amplification in the p53/p73-dependent apoptosis induced by etoposide in HNSCC cells.

[The effect of oral care on natural immune function in the dependent elderly].

While oral care is known to play an important role in the prevention of aspiration pneumonia, its influence on immune function is unclear. In this study, we performed oral care on ten dependent elderly individuals and examined its influence on NK cell activity. The study duration was 12 weeks. During the first six weeks, the subjects cleaned their mouths as usual (control period). In the last six weeks, a dental hygienist performed oral care and denture cleaning three times a week (intervention period). The outcome measure was the quantity of oral bacteria and the detection rate of the microbe in connection with an opportunistic infection and NK cell activity before and after the intervention. Samples were collected at three time points: the beginning of the study, after the control period and before the intervention period (i. e., after six weeks), and after the intervention period (i. e., after 12 weeks). There was no significant difference in the number of oral bacteria between the control period and the intervention period. In contrast, while NK cell activity did not change during the control period, activity increased during the intervention period (p = 0.03). This finding suggests that rather than decreasing the number of oral bacteria, oral care increases NK cell activity.

Inhibition of Streptococcus mutans biofilm formation by Streptococcus salivarius FruA.

The oral microbial flora consists of many beneficial species of bacteria that are associated with a healthy condition and control the progression of oral disease. Cooperative interactions between oral streptococci and the pathogens play important roles in the development of dental biofilms in the oral cavity. To determine the roles of oral streptococci in multispecies biofilm development and the effects of the streptococci in biofilm formation, the active substances inhibiting Streptococcus mutans biofilm formation were purified from Streptococcus salivarius ATCC 9759 and HT9R culture supernatants using ion exchange and gel filtration chromatography. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry analysis was performed, and the results were compared to databases. The S. salivarius HT9R genome sequence was determined and used to indentify candidate proteins for inhibition. The candidates inhibiting biofilms were identified as S. salivarius fructosyltransferase (FTF) and exo-beta-d-fructosidase (FruA). The activity of the inhibitors was elevated in the presence of sucrose, and the inhibitory effects were dependent on the sucrose concentration in the biofilm formation assay medium. Purified and commercial FruA from Aspergillus niger (31.6% identity and 59.6% similarity to the amino acid sequence of FruA from S. salivarius HT9R) completely inhibited S. mutans GS-5 biofilm formation on saliva-coated polystyrene and hydroxyapatite surfaces. Inhibition was induced by decreasing polysaccharide production, which is dependent on sucrose digestion rather than fructan digestion. The data indicate that S. salivarius produces large quantities of FruA and that FruA alone may play an important role in multispecies microbial interactions for sucrose-dependent biofilm formation in the oral cavity.

Effect of the reclining position in patients after oral tumor surgery.

OBJECTIVES: This study investigated the effects of reclining position on swallowing functions in the patients who had undergone surgery for oral tumors. SUBJECTS AND METHODS: 53 Patients after oral tumor surgery who had undergone the videofluoroscopic swallowing study in the sitting position and reclining position were sampled. We measured Oral transit time (OTT), Pharyngeal transit time (PTT), Penetration-Aspiration scale, Locating of the leading edge of bolus at swallow onset, Route of bolus transit and Transit velocity. RESULTS: Laryngeal penetration and aspiration were less likely to occur in the reclining position. The mean OTT was significantly shorter in the reclining position than in the sitting position. The leading edge of bolus at swallow onset was significantly lower in the reclining position than in the sitting position. The bolus went along the posterior wall of the pharynx in more patients in the reclining position. The mean PTT and the transit velocity did not significantly differ between the reclining position and the sitting position. CONCLUSION: This study suggested that, in patients after surgery on oral tumors, the reclining position is a useful posture to aid oral transit and to readily prevent aspiration and laryngeal penetration.

Effects of mucosal care on oral pathogens in professional oral hygiene to the elderly.

To elucidate the effects of mucosal care on colonization of mutans streptococci and infection of Candida species, we randomized 50 functionally dependent nursing home residents, mean age 76.1 (±7.8 years), 76% females, to receive oral professional care with or without mucosal care. During 12 months of follow-up, significant reduction in the numbers of mutans streptococci in saliva and dental plaque samples were observed in both treatment groups. However, there was no reduction in the numbers of mutans streptococci in tongue samples in the non-mucosal group (p=0.150). In the comparison between the treatment with and without mucosal care, the treatment with mucosal care showed a significant inhibition effects on numbers of nutans streptococci on the tongue surface at 6 months after the treatment as compared with the treatment without mucosal care (p=0.043). Mucosal care was not associated with any significant reduction in the rate of opportunistic infection with Candida species in any of the three sites. However, more subjects without mucosal care had Candida infection in dental plaque during the follow-up (p=0.046). Professional dental care with (versus without) mucosal care had no effect on mutans streptococci colonization in saliva or plaque. Lack of mucosal care seemed to be associated with higher rates opportunistic infections with Candida species.

The effects of polyunsaturated fatty acids and their metabolites on osteoclastogenesis in vitro.

Bone homeostasis is maintained by active remodeling through the balance between resorption (by osteoclasts) and synthesis (by osteoblasts). In this study, we examined the effects of polyunsaturated fatty acids (PUFAs) and their metabolites on sRANKL-induced differentiation of bone marrow-derived macrophages (BMMs) into osteoclasts in vitro. Docosahexaenoic acid (DHA) strongly inhibited osteoclastogenesis; however, dihomo-gamma-linolenic acid (DGLA), arachidonic acid (AA) and eicosapentaenoic acid (EPA) enhanced it. The enhancement effect of PUFAs on osteoclastogenesis was mediated predominantly by cyclooxygenase (COX) products, because the effect was inhibited by a COX inhibitor. It was also found that COX products of PUFAs, prostaglandin E(1), E(2), and E(3), clearly increased in osteoclastogenesis. The inhibitory effect of DHA on osteoclastogenesis was reversed by treatment with a lipoxygenase (LOX) inhibitor. Furthermore, resolvin D1, a LOX product of DHA, significantly inhibited osteoclastogenesis. Quantitative analysis of specific mRNA levels revealed that DHA-mediated attenuation of osteoclastogenesis might be due to a decrease in DC-STAMP expression. These results suggested that the effect of DHA on osteoclastogenesis is, at least in part, mediated by lipoxygenase products. This study showed a distinct mechanism of the effect of PUFAs on osteoclastogenesis and will provide evidence for therapeutic treatment with DHA in osteoporotic patients.

E2F-1-deficient NOD/SCID mice developed showing decreased saliva production.

The non-obese diabetic mouse (NOD) is the most characterized model used to study insulin-dependent type 1 diabetes mellitus (IDDM) and Sjoögren's syndrome (SS). In a previous report, we found NOD.E2f1(-/-) mice show a greater progressive development to IDDM and SS compared to NOD mice. Our previous data indicated a progressive decrease in regulatory T cells (CD4(+)CD25(+)) and a decrease in the systemic secretion systems for insulin, and saliva was associated with the progression of IDDM and SS. Therefore, to define the mechanism of early-onset IDDM SS in E2F-1 deficient NOD mice required further investigation by producing E2F-1 deficient NOD/SCID mice in which the T and B cells do not develop. The purpose here was to analyze the essential function of the E2F-1 molecule in the development of IDDM and SS; and the dysfunction of the pancreas islet and salivary gland in the NOD background using NOD/SCID mice. We produced NOD/SCID.E2f1(-/-) mice using homologous recombination; determined diabetes development; measured saliva and insulin production; and performed a histological analysis. The deficient mice showed a decreasing volume of saliva; no infiltration of lymphocytes into salivary glands; no development of diabetes; and no protein localization of FGFR-2b in the ducts of the salivary gland that regulates submandibular gland proliferation and morphogenesis. Therefore, we considered a deficiency in E2F-1 induces a decrease in regulatory T cells and an increase in auto-reactive T cells; however, the E2F-1 deficiency is not associated with T and B cells-independent dysfunction of pancreatic beta cell in insulin secretion. Further, the E2F-1 deficiency is associated with T and B cells-independent dysfunction of the salivary gland exhibits a decrease in saliva production volume. We suggest E2F-1 may be also associated with the differentiation of exocrine cells in the duct where FGFR-2b is expressed in the salivary gland. The E2F-1 deficient NOD/SCID mouse model is useful for showing the development of the salivary gland; and is also useful for various experiments in humanized mice.

Association of caspase-8 mutation with chemoresistance to cisplatin in HOC313 head and neck squamous cell carcinoma cells.

Caspase-8 is a critical upstream mediator of apoptosis in the death receptor pathway. However, the relationship between caspase-8 mutation and chemosensitivity remain unclear in head and neck squamous cell carcinoma (HNSCC) carrying p53 mutation. In this study, we identified a caspase-8 nonsense mutation, accompanied by the loss of the second allele, in a drug-resistant HOC313 HNSCC cell line. The nonsense mutation (R68X) leads to truncation of all defined functional domains. Reconstitution of caspase-8 by stable transfection of wild-type caspase-8 sensitized the cells to cisplatin-, but not etoposide-induced apoptosis. Consistent with this, cisplatin, but not etoposide, induced TNF-alpha and TRAIL mRNA in caspase-8 reconstituted HOC313 cells, accompanied by activation of the reconstituted caspase-8 and its downstream caspase-3. These results indicate that the loss of caspase-8 plays an important role in acquisition of chemoresistance to cisplatin in HOC313 cells.

The effect of lateral shift of cricoid cartilage on pharyngeal swallowing.

Head rotation is widely used as one of the postural techniques for dysphagic patients. However, it cannot be used for patients with severe limitations to the range of motion of the neck. The purpose of this study was to determine the effect of applying pressure to the cricoid while swallowing and to explore the possibility of this maneuver as an alternative to head rotation. The swallowing function of 12 volunteers was examined with videofluorography under nine conditions: neutral, head rotated to the right, head rotated to the left, applying pressure to the cricoid on the right side at 5, 10, and 15 N, and applying pressure to the cricoid on the left side at 5, 10, and 15 N. To examine the effect of this maneuver on pharyngeal swallowing, the laterality of bolus flow was evaluated using 3 ml barium thin liquid from the posterior-anterior (P-A) view. The "pressing-cricoid" maneuver significantly altered the laterality of the bolus flow; the bolus flow was shifted to the side opposite to which the pressure was applied. The results of these changes were similar to those achieved with head rotation. These results demonstrated that the "pressing-cricoid" maneuver changes the bolus flow. This maneuver may have therapeutic value for the treatment of dysphagic patients as an alternative to head rotation.

The role of anti-PAc (361-386) peptide SIgA antibody in professional oral hygiene of the elderly.

OBJECTIVE: Measurement of salivary IgA antibody (PAc-peptide antibody, PPA) to amino acid residues 361-386 of Streptococcus mutans PAc, which possess a multiple binding motif to various HLA-DR molecules and a B-cell epitope that recognises the inhibiting antibody to S. mutans, is an indicator for the population numbers of mutans streptococci (MS) in human saliva. The purpose of this study was to clarify the role of PPA in infection control of MS after professional oral hygiene care. MATERIALS AND METHODS: Thirty-nine dependently living institutionalised elderly subjects (75.9 +/- 7.5 years; 10 males, 29 females) participated in the study. The measurements of PPA, MS, total streptococci (TS) and lactobacilli (LB) were performed by ELISA and culture techniques from saliva, plaque and tongue samples from the elderly. RESULTS: After treatment using professional oral care, the numbers of MS decreased significantly at 6 months in saliva and tongue samples from the group not having PPA in comparison with the primary data; whereas in the PPA-detected group, a significant decrease in MS number was shown immediately following professional care at 1-12 months in all samples. There was little difference in the numbers of LB at any of the time points. The numbers of TS decreased rapidly in PPA-not detected group in comparison with the PPA-detected group. CONCLUSION: PPA may be more effective for controlling MS number in the oral cavity after professional treatment. The measurement of PPA may be used for preventive instruction to dental caries at the chair side in the clinical setting.