Functional analysis of thermo-sensitive TRPV1 in an aquatic vertebrate, masu salmon (Oncorhynchus masou ishikawae).

Transient receptor potential vanilloid 1 (TRPV1) is mainly expressed in nociceptive primary sensory neurons and acts as a sensor for heat and capsaicin. The functional properties of TRPV1 have been reported to vary among species and, in some cases, the species difference in its thermal sensitivity is likely to be associated with thermal habitat conditions. To clarify the functional properties and physiological roles of TRPV1 in aquatic vertebrates, we examined the temperature and chemical sensitivities of TRPV1 in masu salmon (Oncorhynchus masou ishikawae, Om) belonging to a family of salmonids that generally prefer cool environments. First, behavioral experiments were conducted using a video tracking system. Application of capsaicin, a TRPV1 agonist, induced locomotor activities in juvenile Om. Increasing the ambient temperature also elicited locomotor activity potentiated by capsaicin. RT-PCR revealed TRPV1 expression in gills as well as spinal cord. Next, electrophysiological analyses of OmTRPV1 were performed using a two-electrode voltage-clamp technique with a Xenopus oocyte expression system. Heat stimulation evoked an inward current in heterologously expressed OmTRPV1. In addition, capsaicin produced current responses in OmTRPV1-expressing oocytes, but higher concentrations were needed for its activation compared to the mammalian orthologues. These results indicate that Om senses environmental stimuli (heat and capsaicin) through the activation of TRPV1, and this channel may play important roles in avoiding environments disadvantageous for survival in aquatic vertebrates.

Lactobacillus gasseri PA-3 directly incorporates purine mononucleotides and utilizes them for growth.

Lactococcus lactis has been reported unable to directly incorporate mononucleotides but instead requires their external dephosphorylation by nucleotidases to the corresponding nucleosides prior to their incorporation. Although Lactobacillus gasseri PA-3 (PA-3), a strain of lactic acid bacteria, has been found to incorporate purine mononucleotides such as adenosine 5'-monophosphate (AMP), it remains unclear whether these bacteria directly incorporate these mononucleotides or incorporate them after dephosphorylation to the corresponding nucleosides. This study evaluated whether PA-3 incorporated radioactively-labeled mononucleotides in the presence or absence of the 5'-nucleotidase inhibitor α,ß-methylene ADP (APCP). PA-3 took up 14C-AMP in the presence of APCP, as well as incorporating 32P-AMP. Furthermore, radioactivity was detected in the RNA/DNA of bacterial cells cultured in the presence of 32P-AMP. Taken together, these findings indicated that PA-3 incorporated purine mononucleotides directly rather than after their dephosphorylation to purine nucleosides and that PA-3 utilizes these purine mononucleotides in the synthesis of RNA and DNA. Although additional studies are required to identify purine mononucleotide transporters in PA-3, this study is the first to show that some lactic acid bacteria directly incorporate purine mononucleotides and use them for growth.

Species-dependent patterns of incorporation of purine mononucleotides and nucleosides by lactic acid bacteria.

Although most lactic acid bacteria do not directly incorporate purine nucleotides, the strain Lactobacillus gasseri PA-3 was found to incorporate purine mononucleotides. To determine whether the direct uptake of purine mononucleotides is dependent on the species or strain of lactic acid bacteria, incorporation of purine mononucleotides was assessed in L. gasseri, Lactcoccus lactis sbsp. lactis, Streptococcus thermophilus and other species of lactic acid bacteria. Each bacterial strain was incubated with 32P-AMP or 14C-adenosine and the incorporation of each purine was evaluated by measuring their radioactivity. All investigated strains of L. gasseri incorporated 32P-AMP, whereas strains of S. thermophilus and most strains of L. lactis did not. Incorporation of 32P-AMP into strains of Pediococcus was dependent on the strain or species of that genus of bacteria. All investigated strains, except for one strain of L. gasseri, incorporated 14C-adenosine, with S. thermophilus, L. lactis and Pediococcus generally displaying greater incorporation of 14C-adenosine than L. gasseri. Although most lactic acid bacteria such as S. thermophiles and L. lactis do not incorporate purine mononucleotides, some species such as L. gasseri directly incorporate purine mononucleotides. These findings indicate that the preferential incorporation of purine mononucleotides or nucleosides by lactic acid bacteria is dependent on the species or strain.

Lactobacillus gasseri PA-3, but not L. gasseri OLL2996, reduces the absorption of purine nucleosides in rats.

Lactobacillus gasseri PA-3 (PA-3) is a bacterial strain with a strong ability to degrade purine nucleosides. We previously showed that PA-3 incorporates purines in vitro and that oral administration of PA-3 and purines to rats attenuated their absorption of purines. It remains unclear whether these effects of PA-3 depend on bacterial strains. This study therefore compared the abilities of PA-3 and another bacterial strain of L. gasseri, OLL2996, which has shown decreased ability to degrade purine nucleosides in vitro, to incorporate purine nucleosides and to inhibit the absorption of purines fed to rats. Each bacterial strain was incubated in the presence of 14C-adenosine or 14C-inosine and the incorporation of each purine was evaluated by measuring their radioactivity. In vivo, rats were fed 14C-labeled purines along with PA-3 or OLL2996 and the absorption of these 14C-labeled purines was evaluated by analyzing radioactivity of blood samples. PA-3 incorporated about twice as much 14C-adenosine and 14C-inosine as OLL2996. The elevation of radioactivity levels in blood was 10-20% lower in rats treated with PA-3 than in control rats, after feeding with both 14C-adenosine and 14C-inosine as purines. In contrast, treatment with OLL2996 did not have statistically significant effects on radioactivity compared with the control group. These results indicate that the magnitude of bacterial inhibition of purine absorption is dependent on bacterial strain, correlating at least partly with the ability to incorporate and degrade purines.

Lactobacillus gasseri PA-3 utilizes the purines GMP and guanosine and decreases their absorption in rats.

Excessive intake of purine-rich foods elevates serum uric acid levels, making it a risk factor for hyperuricemia. We hypothesized that lactic acid bacteria ingested with food might utilize purines and contribute to their decreased absorption in the intestines, thereby preventing hyperuricemia. We previously reported that Lactobacillus gasseri PA-3 (PA-3) incorporates adenosine/inosine and related purines and that oral ingestion of PA-3 reduced the absorption of these purines in rats. However, it is unclear whether PA-3 also decreases the absorption of other purines, such as guanosine 5'-monophosphate (GMP) and guanosine. This study investigated whether PA-3 incorporates GMP and guanosine and reduces their absorption in rats. PA-3 incorporated both purines, with 14C-GMP uptake being greater than that of 14C-guanosine. Radioactivity in rat blood was significantly lower 30, 45, and 60 minutes after administration of 14C-GMP plus PA-3 than after administration of 14C-GMP alone and was significantly lower 15 minutes after administration of 14C-guanosine plus PA-3 than after administration of 14C-guanosine alone. PA-3 incorporates GMP and guanosine in vitro. Oral administration of PA-3 with GMP and guanosine reduces the intestinal absorption of these purines in vivo. These findings, together with those of previous studies, indicate that PA-3 reduces the absorption of major purines contained in foods. PA-3 may also attenuate the excessive absorption of dietary purines in humans, protecting these individuals against hyperuricemia.

Prognostic value of tumor-infiltrating lymphocytes differs depending on histological type and smoking habit in completely resected non-small-cell lung cancer.

BACKGROUND: T-cell infiltration in tumors has been used as a prognostic tool in non-small-cell lung cancer (NSCLC). However, the influence of smoking habit and histological type on tumor-infiltrating lymphocytes (TILs) in NSCLC remains unclear. PATIENTS AND METHODS: We evaluated the prognostic significance of TILs (CD4+, CD8+, CD20+, and FOXP3+) according to histological type and smoking habit using automatic immunohistochemical staining and cell counting in 218 patients with NSCLC. RESULTS: In multivariate survival analyses of clinical, pathological, and immunological factors, a high ratio of FOXP3+ to CD4+ T cells (FOXP3/CD4) [hazard ratio (HR): 4.46, P < 0.01 for overall survival (OS); HR: 1.96, P < 0.05 for recurrence-free survival (RFS)] and a low accumulation of CD20+ B cells (HR: 2.45, P = 0.09 for OS; HR: 2.86, P < 0.01 for RFS) were identified as worse prognostic factors in patients with adenocarcinoma (AD). In non-AD, a low number of CD8+ T cells were correlated with an unfavorable outcome (HR: 7.69, P < 0.01 for OS; HR: 3.57, P < 0.02 for RFS). Regarding smoking habit in AD, a high FOXP3/CD4 ratio was poorly prognostic with a smoking history (HR: 5.21, P < 0.01 for OS; HR: 2.38, P < 0.03 for RFS), whereas a low accumulation of CD20+ B cells (HR: 4.54, P = 0.03 for OS; HR: 2.94, P < 0.01 for RFS) was confirmed as an unfavorable factor in non-smokers with AD. CONCLUSIONS: A low number of CD8+ T cells in non-AD, a high FOXP3/CD4 ratio in smokers with AD, and a low number of CD20+ B cells in non-smokers with AD were identified as independent unfavorable prognostic factors in resected NSCLC. Evaluating the influence of histological type and smoking habit on the immunological environment may lead to the establishment of immunological diagnosis and appropriate individualized immunotherapy for NSCLC.

Effects of orthognathic surgery on psychological status of patients with jaw deformities.

The purpose of this study was to determine the effect of orthognathic surgery on psychological status. The subjects were 119 patients (38 males and 81 females, mean age 25.5±9.4 years) who underwent orthognathic surgery. They were divided into class III (84 patients), class II (20 patients), and class I (15 patients) groups according to the anteroposterior skeletal pattern, and they were also divided into an asymmetry group (51 patients) and a symmetry group (68 patients). We assessed psychological status using the Minnesota Multiphasic Personality Inventory (MMPI) before surgery and at more than 6 months after surgery. The MMPI scores for the depression, hysteria, psychasthenia, and social introversion scales were significantly higher than standard values before surgery, and the hypomania scale significantly lower. The cannot say scale, depression scale, and hysteria scale decreased significantly after surgery. A comparison of MMPI scores among the groups showed the depression scale in the class III group to be higher than those in the class I and II groups; there was no significant difference between the asymmetry and symmetry groups. In conclusion, orthognathic surgery has a positive influence on the psychological status of patients with jaw deformities, especially patients with skeletal class III malocclusion.

Impairment of host resistance to helminthes with age in murine small intestine.

Age-associated alterations of Th2 immune responses against nematode parasites are largely unknown. We investigated primary and memory responses against two types of gastrointestinal nematode parasites, Heligmosomoides polygyrus (Hp) and Nippostrongylus brasiliensis (Nb), in aged mice. The small intestinal gene expression of Th2 cytokines was almost unchanged after primary (Nb and Hp) and secondary infection (Hp) in aged mice in contrast to strongly increased small intestinal gene expression of Th2 cytokines in young (3-month-old) mice. Mucus production decreased (Nb), and worm expulsion was impaired (Nb and Hp) compared with the young mice. Immunofluorescent staining revealed that after Hp infection, the number of alternatively activated macrophages, which are induced by Th2 cytokines, was lower in the aged mice. On the other hand, the number of CD4(+) T cells recruited to the worm cysts was normal compared with the young mice. These results suggest that migration of CD4(+) T cells to the host-parasite interface is not affected by ageing. Alterations in Th2 immune responses in aged mice might be due to inappropriate or insufficient activation of CD4(+) T cells in the submucosa.

Tuberculosis in pregnancy:a challenging differential diagnosisfor inflammatory bowel disease.

OBJECTIVE: To describe a case of tuberculosis with intestinal variant in a pregnant woman in the 17th week of pregnancy. DESIGN: Case report. SETTING: Department of Obstetrics, Federal University of São Paulo (UNIFESP), São Paulo-SP, Brazil. CONCLUSION: Tuberculosis is a public health problem that concerns many countries in the world. It was declareda public emergency by the World Health Organization in 2005. Its presence during pregnancy brings maternal risk and fetal impairment if not treated quickly and properly. The intestinal variant is not the most common form of the disease and may be confused with inflammatory bowel diseases, especially Crohns disease. Knowledge of the specific characteristics, combined with a detailed medical history and appropriate diagnostic methods can make all the difference in gestational prognosis. We report the case of a pregnant woman who wrongly underwent treatment for inflammatory bowel disease at another service. After admission to our university hospital, fruitful diagnostic clarification was achieved and the patient was diagnosed and treated for tuberculosis. We describe the details of the investigation and, in particular, review the main characteristics in the literature for differentiating the two diseases.

Effects of orthognathic surgery on pharyngeal airway and respiratory function during sleep in patients with mandibular prognathism.

The aim of this study was to determine changes in overnight respiratory function and craniofacial and pharyngeal airway morphology following orthognathic surgery. The subjects were 40 patients in whom mandibular prognathism was corrected by orthognathic surgery: a one-jaw operation in 22 patients and a two-jaw operation in 18 patients. Morphological changes were studied using cone beam computed tomography immediately before surgery and at more than 6 months after surgery, and the apnoea-hypopnoea index (AHI) was measured with a portable polysomnography system. Pharyngeal airway volume was decreased significantly after surgery, especially in the one-jaw operation group. AHI was not changed significantly after surgery in either group, although AHI in one patient in the one-jaw operation group was increased to 19 events/h. There was no significant change in pharyngeal airway morphology in that patient, but he was obesity class 1 and was 54 years old. In conclusion, some patients who are obese, have a large amount of mandibular setback, and/or are of relatively advanced age may develop sleep-disordered breathing after mandibular setback; a two-jaw operation should therefore be considered in skeletal class III patients who have such risks because it decreases the amount of pharyngeal airway space reduction caused by mandibular setback surgery.

IgA pemphigus with paraneoplastic pemphigus-like clinical features showing IgA antibodies to desmoglein 1/3 and desmocollin 3, and IgG and IgA antibodies to the basement membrane zone.

A 79-year-old Japanese woman presented with severe recalcitrant erosions on her oral mucosa, resembling paraneoplastic pemphigus. Using indirect immunofluorescence, we detected IgA antibodies against the cell surface, and both IgG and IgA antibodies against the basement membrane zone. Immunoblotting showed that the IgG antibodies reacted weakly with bullous pemphigoid 230 and periplakin, whereas the IgA antibodies did not react with any antigen. IgA antibodies to both desmoglein (Dsg)1 and Dsg3 were detected by ELISA. IgA antibodies to desmocollin (Dsc)3 were also detected by using cDNAs for human Dsc1-3 transfected into COS-7 cells. Despite treatment with oral prednisolone, high-dose intravenous immunoglobulin and double-filtration plasmapheresis, the skin lesions remained active, and the patient died from bronchiolitis obliterans-like respiratory failure. Despite extensive investigations and postmortem examination, no underlying neoplasms were found. The complex immunopathological findings probably played an important role in the development of the patient's unusual clinical features.

Inflammatory histopathogenesis of nasopalatine duct cyst: a clinicopathological study of 41 cases.

OBJECTIVE: The aim of this study is to characterize immunohistochemical profiles of lining epithelia of nasopalatine duct cyst (NPC) as well as to correlate those findings with their clinicopathological features to understand the histopathogenesis of NPC. MATERIALS AND METHODS: Forty-one surgical specimens from NPC were examined for clinical profiles and expression of keratin-7, 13, MUC-1, and P63 by immunohistochemistry, compared to radicular cyst (RC) and maxillary sinusitis. RESULTS: Nasopalatine duct cyst was clinically characterized by male predominant occurrence: 44% of the cases involved tooth roots, and 70% with inflammatory backgrounds. Lining epithelia of NPCs without daughter cysts were immunohistochemically distinguished into three layers: a keratin 7-positive (+) ciliated cell layer in the surface, a keratin-13+ middle layer, and a MUC-1+/P63+ lower half, indicating that they were not respiratory epithelia, and the same layering pattern was observed in RC. However, those immunolocalization patterns of the main cyst lining with daughter cyst were exactly the same as those of daughter cyst linings as well as duct epithelia of mucous glands. CONCLUSIONS: Two possible histopathogenesis of NPC were clarified: one was inflammatory cyst like RC and the other was salivary duct cyst-like mucocele.

Evaluation of bone volume changes after sinus floor augmentation with autogenous bone grafts.

The aim of this study was to establish an objective method for quantitative evaluation of bone volume change after sinus augmentation. 11 sinuses in 9 patients were evaluated by computed tomography images taken before treatment (T0), and 3 months (T1) and at least 1 year (T2) after sinus augmentation. Based on the 3D digital subtraction technique, augmented bone images were extracted and bone volumes were calculated from voxel numbers of the extracted images. The mean augmented bone volumes at T1 and T2 were 2.46 cm3 and 1.85 cm3, respectively. These bone volume changes were statistically significant and the mean bone volume change ± SE was -24.8% ± 6.1%. Loss of augmented bone was observed in all except one of the patients. The correlation coefficient between bone volume change and elapsed time was -0.64, which was statistically significant and indicated that bone resorption progressed with elapse of time after sinus augmentation. The authors' method of analysis enabled visualization of augmented bone and objective assessment of bone volume change. Within the limited number of cases, the present investigation demonstrated a significant decrease in augmented bone volume between 3 and 23 months after surgery.

Adenomyoma associated with high level of CA 125 and CA 19-9: case report.

PURPOSE OF INVESTIGATION: A rare case of increasing CA 125 and CA 19-9 levels increasing in a woman with adenomyoma is described. METHODS: A 39-year-old nullipara woman with CA 125 = 1,796 U/ml and CA 19-9 = 177 U/ml was submitted to abdominal and pelvic MRI, gastric endoscopy, colonoscopy, hysteroscopy, pelvic Doppler and PET scan. None of the exams revealed any apparent malignant disease. RESULTS: Six months of gonadotropin releasing hormone agonist treatment reduced CA 125 and CA 19-9 levels. However, after contraceptive pill use the markers were again elevated, and a laparoscopic hysterectomy was performed, and normal CA 125 and CA 19-9 levels were achieved. CONCLUSIONS: Adenomyoma may be associated with high levels of CA 125 and CA 19-9.

Changes in oropharyngeal airway and respiratory function during sleep after orthognathic surgery in patients with mandibular prognathism.

The aim of this study was to examine the effects of mandibular setback surgery on pharyngeal airway space and respiratory function during sleep. The subjects were 22 patients in whom mandibular prognathism was corrected by bilateral sagittal split ramus osteotomy; either one jaw or two jaw surgery. Polysomnography was performed before surgery and 6 months after surgery, and the apnea hypopnea index (AHI) and arterial oxygen saturation during sleep were measured to assess respiratory function during sleep. Morphological changes were studied using cephalograms taken immediately before, a few days after and 6 months after surgery. As a control, 10 subjects without sleep-disordered breathing underwent the same examinations. AHI was not changed significantly after surgery, although two patients were diagnosed with mild obstructive sleep apnea (OSA) syndrome after surgery. They were not obese, but the amounts of mandibular setback at surgery were large. In conclusion, a large amount of mandibular setback might inhibit biological adaption and cause sleep-disordered breathing, and it might be better to consider maxillary advance or another technique that does not reduce the airway for patients with skeletal class III malocclusions who have large anteroposterior discrepancy and/or maxillary hypoplasia.

Complication of adenoid cystic carcinoma and sialolithiasis in the submandibular gland: report of a case and its etiological background.

The authors report a case of adenoid cystic carcinoma (ACC) complicated with sialolithiasis of the submandibular gland. The patient was a 43-year-old female with a history of papillotubular carcinoma of the breast almost at the same time. She had noticed a swelling in her sublingual area for 10 years, which was later diagnosed by her dentist to be due to a sialolith in the left submandibular gland. After several years of observation, the patient was referred to have her left submandibular gland, containing the stone, surgically removed with a diagnosis of atrophic sialadenitis. Histopathologically, the submandibular gland was extensively replaced with fibrous granulation tissue, in which there were small but invasive foci of ACC. The present case indicates that ACC could arise in the background of chronic sialadenitis. It is suggested that long-standing sialadenitis cases should be carefully examined to exclude suspicion of malignancy before surgery.

Non-syndromic oligodontia with a novel mutation of PAX9.

Agenesis of the permanent teeth is a congenital anomaly that is frequently seen in humans. Oligodontia is a severe type of tooth agenesis involving 6 or more congenitally missing teeth, excluding the third molars. Previous studies have indicated that mutations in the homeobox gene MSX1, paired domain transcription factor PAX9, and EDA are associated with non-syndromic oligodontia. This study reports a Japanese family (eight of 14 family members affected) with non-syndromic oligodontia who preferentially lacked molar teeth. In this family, a novel frameshift mutation (321_322insG) was identified in the paired domain of PAX9. The frameshift mutation caused altered amino acids in the paired domain and premature termination of translation by 26 amino acids. When transfected into COS-7 cells, the mRNA expression of 321_322insG PAX9 was comparable with that of wild-type PAX9. However, the mRNA of 321_322insG PAX9 was more unstable than that of wild-type PAX9. This mRNA instability caused a marked decrease in protein production, as evaluated by Western blot analysis and immunostaining. These findings suggest that the 321_322insG mutation causes insufficient function of PAX9 protein and haploinsufficiency as a genetic model of familial non-syndromic oligodontia with a PAX9 mutation.

Cone contrast influence on components of the pattern onset/offset VECP.

Transient visual evoked cortical potentials (VECP) were recorded from the scalp of healthy normal trichromats (n = 12). VECPs were elicited by onset/offset presentation of patterned stimuli of two kinds: isochromatic luminance-modulated, and equiluminant red-green modulated, sine wave gratings. The amplitude and latency of the major onset components of the onset/offset VECP were measured and plotted as a function of the logarithm of pooled cone contrast. The early onset components, achromatic C1 and chromatic N1, increase linearly with log contrast, but N1 has a higher contrast gain than C1. The late onset components, achromatic C2 and chromatic N2, have similar contrast gain, and similar response as a function of contrast level: both increase in the low-to-medium range of contrasts and saturate at high contrast levels. In the range of pooled cone contrast tested, C1 and N1 show similar latencies, whilst C2 shows shorter latencies than N2. We suggest that C1 and N1 are generated by the same visual mechanism with high red-green contrast gain and low luminance contrast gain, whilst C2 and N2 are generated by different visual mechanisms.

Clinical study on prognostic factors for autotransplantation of teeth with complete root formation.

Autotransplantation is often performed to replace a missing tooth, but tooth autotransplantation has been reported in fewer teeth with complete root formation than those with incomplete root formation. The aim of this prospective study was to evaluate the factors that affect the prognosis of autotransplantation of teeth with complete root formation. 109 patients with 117 transplants were studied. Of the 117 transplants investigated, 14 (12%) failed during the observation period. The overall 1-year survival rate was 96%; the 5-year survival rate was 84%. The major causes of failure were unsuccessful initial healing and replacement root resorption with periodontal inflammation. Factors significantly associated with unsuccessful transplantation, in single factor analysis, were age 40 years or more, molar tooth as donor, probing pocket depth to 4mm or more, history of root canal treatment, multi-rooted teeth and fixation with sutures. Pocket depth of 4mm or more and history of root canal treatment appeared to increase the risk of unsuccessful transplantation in multivariate analysis. It is suggested that the pocket depth of the donor tooth and history of root canal treatment are related to the healing of paratransplantal tissue and root resorption.

Twelve chromatically opponent ganglion cell types in turtle retina.

The turtle retina has been extensively used for the study of chromatic processing mechanisms. Color opponency has been previously investigated with trichromatic paradigms, but behavioral studies show that the turtle has an ultraviolet (UV) channel and a tetrachromatic visual system. Our laboratory has been working in the characterization of neuronal responses in the retina of vertebrates using stimuli in the UV-visible range of the electromagnetic spectrum. In the present investigation, we recorded color-opponent responses from turtle amacrine and ganglion cells to UV and visible stimuli and extended our previous results that UV color-opponency is present at the level of the inner nuclear layer. We recorded from 181 neurons, 36 of which were spectrally opponent. Among these, there were 10 amacrine (5%), and 26 ganglion cells (15%). Morphological identification of color-opponent neurons was possible for two ganglion cell classes (G17 and G22) and two amacrine cell classes (A22 and A23b). There was a variety of cell response types and a potential for complex processing of chromatic stimuli, with intensity- and wavelength-dependent response components. Ten types of color opponency were found in ganglion cells and by adding previous results from our laboratory, 12 types of opponent responses have been found. The majority of the ganglion cells were R+UVBG- and RG+UVB-color-opponents but there were other less frequent types of chromatic opponency. This study confirms the participation of a UV channel in the processing of color opponency in the turtle inner retina and shows that the turtle visual system has the retinal mechanisms to allow many possible chromatic combinations.