A spike analysis method for characterizing neurons based on phase locking and scaling to the interval between two behavioral events.

Standard analysis of neuronal functions assesses the temporal correlation between animal behaviors and neuronal activity by aligning spike trains with the timing of a specific behavioral event, e.g., visual cue. However, spike activity is often involved in information processing dependent on a relative phase between two consecutive events rather than a single event. Nevertheless, less attention has so far been paid to such temporal features of spike activity in relation to two behavioral events. Here, we propose "Phase-Scaling analysis" to simultaneously evaluate the phase locking and scaling to the interval between two events in task-related spike activity of individual neurons. This analysis method can discriminate conceptual "scaled"-type neurons from "nonscaled"-type neurons using an activity variation map that combines phase locking with scaling to the interval. Its robustness was validated by spike simulation using different spike properties. Furthermore, we applied it to analyzing actual spike data from task-related neurons in the primary visual cortex (V1), posterior parietal cortex (PPC), primary motor cortex (M1), and secondary motor cortex (M2) of behaving rats. After hierarchical clustering of all neurons using their activity variation maps, we divided them objectively into four clusters corresponding to nonscaled-type sensory and motor neurons and scaled-type neurons including sustained and ramping activities, etc. Cluster/subcluster compositions for V1 differed from those of PPC, M1, and M2. The V1 neurons showed the fastest functional activities among those areas. Our method was also applicable to determine temporal "forms" and the latency of spike activity changes. These findings demonstrate its utility for characterizing neurons.NEW & NOTEWORTHY Phase-Scaling analysis is a novel technique to unbiasedly characterize the temporal dependency of functional neuron activity on two behavioral events and objectively determine the latency and form of the activity change. This powerful analysis can uncover several classes of latently functioning neurons that have thus far been overlooked, which may participate differently in intermediate processes of a brain function. The Phase-Scaling analysis will yield profound insights into neural mechanisms for processing internal information.

Targeting ID2 expression triggers a more differentiated phenotype and reduces aggressiveness in human salivary gland cancer cells.

Inhibitors of DNA-binding (ID) proteins are negative regulators of basic helix-loop-helix transcription factors and generally stimulate cell proliferation and inhibit differentiation. We previously determined that ID1 was highly expressed in aggressive salivary gland cancer (SGC) cells in culture. Here, we show that ID2 is also expressed in aggressive SGC cells. ID2 knockdown triggers important changes in cell behavior, that is, it significantly reduces the expression of N-cadherin, vimentin and Snail, induces E-cadherin expression and leads to a more differentiated phenotype exemplified by changes in cell shape. Moreover, ID2 knockdown almost completely suppresses invasion and the expression of matrix metalloproteinase 9. In conclusion, ID2 expression maintains an aggressive phenotype in SGC cells, and ID2 repression triggers a reduction in cell aggressiveness. ID2 therefore represents a potential therapeutic target during SGC progression. ID proteins are negative regulators of basic helix-loop-helix transcription factors and generally stimulate cell proliferation and inhibit differentiation. ID2 knockdown triggers important changes in cell behavior, that is, it significantly reduces the expression of N-cadherin, vimentin and Snail, induces E-cadherin expression and leads to a more differentiated phenotype exemplified by changes in cell shape. ID2 therefore represents a potential therapeutic target during SGC progression.

Hierarchy between forelimb premotor and primary motor cortices and its manifestation in their firing patterns.

Though hierarchy is commonly invoked in descriptions of motor cortical function, its presence and manifestation in firing patterns remain poorly resolved. Here we use optogenetic inactivation to demonstrate that short-latency influence between forelimb premotor and primary motor cortices is asymmetric during reaching in mice, demonstrating a partial hierarchy between the endogenous activity in each region. Multi-region recordings revealed that some activity is captured by similar but delayed patterns where either region's activity leads, with premotor activity leading more. Yet firing in each region is dominated by patterns shared between regions and is equally predictive of firing in the other region at the single-neuron level. In dual-region network models fit to data, regions differed in their dependence on across-region input, rather than the amount of such input they received. Our results indicate that motor cortical hierarchy, while present, may not be exposed when inferring interactions between populations from firing patterns alone.

Targeting Id1 reduces proliferation and invasion in aggressive human salivary gland cancer cells.

BACKGROUND: Salivary gland cancer (SGC) is one of the common malignancies of the head and neck area. It develops in the minor and major salivary glands and sometimes metastasizes to other organs, particularly to the lungs. Inhibitors of differentiation (Id) proteins are negative regulators of basic helix-loop-helix transcription factors that control malignant cell behavior and tumor aggressiveness in many tissues. In this study, our goal was to determine the potential role of Id proteins, particularly Id1, during human SGC cell progression. METHODS: We first determined the expression levels of Id1 and Id2 in four SGC cell lines: two adenocarcinoma of the salivary gland (HSG and HSY) and two adenoid cystic carcinoma (ACC2 and ACCM) cell lines. We then used constructs that expressed antisense cDNAs to Id1 or Id2 to knockdown the expression of these proteins in cell lines where they were highly expressed, and determined the effects of the knockdown on cell proliferation, migration and invasion. RESULTS: Id1 mRNA and protein were detectable in all cell lines, and expression of Id2 was variable, from absent to high. The ACC2 and ACCM cell lines expressed both Id1 and Id2, but Id1 was expressed at a higher level in the more aggressive ACCM cell line in comparison to ACC2 cells as confirmed by Id1 promoter-reporter assays. We therefore focused on the ACCM cells for the remainder of the study. We found that proliferation and invasiveness of ACCM cells were strongly reduced after Id1 knockdown whereas Id2 suppression had only a slight effect. Results of scratch and colony formation assays also confirmed that ACCM cell aggressiveness was significantly reduced upon Id1 knockdown. Finally, this knockdown resulted in reduced c-myc and enhanced cyclin-dependent kinase inhibitor p21 expression. CONCLUSIONS: These results demonstrate that Id1 plays an important role in the control of human SGC cell aggressiveness and suggest a potential role as a marker of diagnosis, prognosis and progression of SGCs. Id1 suppression could represent a novel and effective approach for the treatment of salivary gland cancer.

[Palliative care during chemotherapy for advanced colon cancers].

BACKGROUND: The median survival time following chemotherapy for unresectable metastatic colorectal cancer (mCRC) is approximately 2 years. Although palliative care during the chemotherapy period is very important, it has not been reported in detail. PATIENTS AND METHODS: Information on the palliative care of 110 patients with Stage IV mCRC, who were treated from September 2007 to March 2011, was retrospectively examined. RESULTS: Following an explanation of their recurrence or metastases of mCRC, all the patients received mental care from nurses or psychiatrists. They also needed care to prevent the side effects of chemotherapy. Some patients experienced pain associated with tumor growth. Thus, they required NSAIDs or opioids to reduce the cancer-related pain. After they could not be taken chemotherapy, 87.5% of these patients consulted medical social workers to discuss where they would live. CONCLUSIONS: The patients required palliative care depending on the duration of chemotherapy for mCRC. Thus, we believe that palliative care is an important part of treatment for advanced cancer.

Enhanced epithelial-mesenchymal transition-like phenotype in N-acetylglucosaminyltransferase V transgenic mouse skin promotes wound healing.

N-Acetylglucosaminyltransferase V (GnT-V) catalyzes the ß1,6 branching of N-acetylglucosamine on N-glycans. GnT-V expression is elevated during malignant transformation in various types of cancer. However, the mechanism by which GnT-V promotes cancer progression is unclear. To characterize the biological significance of GnT-V, we established GnT-V transgenic (Tg) mice, in which GnT-V is regulated by a ß-actin promoter. No spontaneous cancer was detected in any organs of the GnT-V Tg mice. However, GnT-V expression was up-regulated in GnT-V Tg mouse skin, and cultured keratinocytes derived from these mice showed enhanced migration, which was associated with changes in E-cadherin localization and epithelial-mesenchymal transition (EMT). Further, EMT-associated factors snail, twist, and N-cadherin were up-regulated, and cutaneous wound healing was accelerated in vivo. We further investigated the detailed mechanisms of EMT by assessing EGF signaling and found up-regulated EGF receptor signaling in GnT-V Tg mouse keratinocytes. These findings indicate that GnT-V overexpression promotes EMT and keratinocyte migration in part through enhanced EGF receptor signaling.

Epidural cooling minimizes spinal cord injury after aortic cross-clamping through induction of nitric oxide synthase.

BACKGROUND: By using a U-shaped lumen catheter, the authors examined the effects of epidural cooling on spinal cord injury after aortic cross-clamping (ACC), with a focus on changes of spinal cord blood flow and expression of inducible nitric oxide synthase. METHODS: Sixteen pigs were randomized into two groups: Control group (n = 8) or Cooling group (n = 8). In the latter, epidural cooling started at 30 min (baseline) before 45 min of ACC and persisted for the next 30 min of reperfusion period. Spinal cord blood flow and somatosensory-evoked potentials were assessed during peri-ACC period. At 48 h, we evaluated hind limb function by using Tarlov score and expression of inducible nitric oxide synthase on spinal cord using immunohistochemistry. RESULTS: After ACC, spinal cord blood flow dropped to a similar extent in both groups. During the reperfusion period, spinal cord blood flow increased up to 113% (103-124%), median (interquartile range), level transiently and decreased to 32% (22-47%) level versus baseline in the Control group, whereas it increased and remained at 92% (86-97%) level in the Cooling group. Simultaneously, somatosensory-evoked potentials showed that onset of loss time was delayed and recovery time was shortened in the Cooling group. Tarlov scores in the Cooling group were significantly higher and accompanied by normal-appearing motor neurons and significantly greater expression of inducible nitric oxide synthase on spinal cord versus the Control group. CONCLUSIONS: This study shows that epidural cooling during ACC minimized the risk of spinal cord injury, possibly by preventing delayed hypoperfusion and upregulating inducible nitric oxide synthase expression.

Potent antioxidant and radical-scavenging activity of Chenpi--compensatory and cooperative actions of ascorbic acid and citric acid.

Dried peels of Satsuma mandarin (Citrus unshiu Marcov.) have been used as traditional Chinese and Japanese medicine, which are called 'Chenpi'. In our present study, cold and hot water extracts of Chenpi exhibited a strong inhibitory activity against linoleic acid peroxidation and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging activity compared with o-methanol extract. When these extracts were separated into ethanol-soluble(ES) and ethanol-precipitable fractions, the dominant antioxidant and radical-scavenging activities were detected in the ES fractions, which suggests that these antioxidant and radical-scavenging activities are responsible for water-soluble and low-molecular-weight substances. As possible active principles for antioxidant activities in the water extracts, the contents of ascorbic acid and citric acid in these extracts were measured, and the antioxidant and radical-scavenging activities of these substances were assayed at various concentrations. The experimental results indicate that the antioxidant activity against lipid peroxidation in the water extracts is dominantly associated with citric acid, and the DPPH radical-scavenging activity of the water extracts is majorly responsible for ascorbic acid, suggesting a compensatory action of ascorbic acid and citric acid in expression of the antioxidant and radical-scavenging activities of Chenpi.

Mandibular central leiomyosarcoma with high telomerase activity: a case report.

Leiomyosarcoma is a malignant lesion of smooth muscle origin, and rare in the oral region. This report presents an extremely rare case of intraosseous leiomyosarcoma of the mandible. After visiting other general hospital, a 29-year-old man was referred to our hospital because of a pain in the left mandibular region with paresthesia of the left mental region. The left mandibular third molar had already been extracted in another hospital, and a brownish mass occupied the corresponding region. A panoramic radiograph showed osteolytic destruction around the left mandibular angle and ramus. A computed tomography scan and magnetic resonance image revealed perforation of the lingual and buccal cortex of the mandible. A non-epithelial malignant tumor was diagnosed from a biopsy specimen. Immediately, we resected the tumor and reconstructed the titan plate under general anesthesia. A final diagnosis of leiomyosarcoma was made from a surgical specimen based on findings showing a proliferation of hyperchromatic spindle cells, which were positive for the markers α- smooth muscle actin, calponin, HHF35, and desmin. The S-100, epithelial membrane antigen, and cytokeratin markers were negative. The patient had 3 courses of adjuvant chemotherapy after the operation, and showed no evidence of recurrence during the follow-up at the outpatient clinic. However, 2 years after the first operation, lung metastases and local recurrence were detected. Additional chemotherapy was not effective. Finally, the patient died almost 3 years after the first operation.

Characterization of inhibitors of postprandial hyperglycemia from the leaves of Nerium indicum.

Nerium indicum is an India-Pakistan-originated shrub belonging to the oleander family. The ingestion of leaves of N. indicum before a meal is known to effect the lowering of postprandial glucose levels in Type II diabetic patients and this plant is now used as a folk remedy for Type II diabetes in some regions of Pakistan. In the present study, the hot-water extract of N. indicum leaves was found to reduce the postprandial rise in the blood glucose when maltose or sucrose was loaded in rats. It was also found that the extract strongly inhibited alpha-glucosidase, suggesting that the suppression of the postprandial rise in the blood glucose is due to the occurrence of some inhibitors of alpha-glucosidase in the leaves. We, therefore, tried to isolate the active principles from the leaf extract, using alpha-glucosidase-inhibitory activity as the index. Employing Sephadex G-15, silica gel and reversed-phase HPLC, we isolated two active compounds. The UV, mass and NMR spectrometric analyses established that the chemical structures of these compounds are 3-O-caffeoylquinic acid (chlorogenic acid) and its structural isomer, 5-O-caffeoylquinic acid. Both compounds were shown to inhibit alpha-glucosidases in a non-competitive manner. The authentic chlorogenic acid was found to suppress the postprandial rise in the blood glucose in rats and also inhibited the absorption of the glucose moiety from maltose and glucose in the everted gut sac system prepared from rat intestine. These results demonstrate that chlorogenic acid is one of the major anti-hyperglycemic principles present in the leaves of N. indicum. Furthermore, among polyphenol compounds tested, quercetin and catechins were shown to have strong inhibitory activity against alpha-glucosidase.

Stimulation of the Estrogen Axis Induces Epithelial-Mesenchymal Transition in Human Salivary Cancer cells.

BACKGROUND: Salivary gland cancer is a common type of head and neck cancer characterized by occasional deep invasion and lung metastasis. The precise role of sex steroid hormones in salivary gland cancer is unclear. To address this issue, we investigated whether the estrogen axis modulates salivary adenocarcinoma (SAC) and whether hormone therapy can be an effective treatment. MATERIALS AND METHODS: The estrogen receptor (ER) was overexpressed in HSG human SAC cells that lack endogenous ER and the cells were treated with and without 17ß-estradiol (E2). RESULTS: E2 enhanced malignant phenotypes. Moreover, E2 treatment reduced E-cadherin expression, while increasing that of N-cadherin, vimentin, and inhibitor of differentiation 1 proteins that are associated with the epithelial-mesenchymal transition. Cell invasion was enhanced through activation of matrix metalloproteinase-9. CONCLUSION: These results indicate that hormone therapy used in breast cancer may also be effective for ER-positive SAC.

The Contribution of Dental Implants to Functional Artificial Restoration After Treatment of Oral Cancer.

BACKGROUND/AIM: The aim of this study was to evaluate dental implants with regard to artificial restoration of oral function and quality of life in patients with oral cancer. PATIENTS AND METHODS: We examined 134 implants in 41 patients who had undergone jawbone resection as treatment for oral cancer. The patients were aged 44-89 (mean=61.5) years, and the male to female ratio was 27:14. RESULTS: The 5-year implant success rate was 91.0%. Of the 12 unsuccessful implants, four were embedded on bone grafts with skin flaps, four were embedded on skin flaps using muscle, and four were embedded after peripheral resection. Of the 41 patients, 11 received radiation, but exposure to radiation was not associated with implant loss. The level of satisfaction on the visual analog scale before development of oral cancer was set at 100 mm. Satisfaction fell to 47.0 mm after primary treatment, but recovered to 82.6 mm after implant therapy. CONCLUSION: Patient satisfaction after implant therapy was high, and the implants resulted in improved quality of life. A high proportion of cases involving use of skin flaps resulted in implant loss. Constructing an immobile mucous membrane by replacement of a skin flap with a skin graft may facilitate self-maintenance of implants.

ID1 Controls Aggressiveness of Salivary Gland Cancer Cells via Crosstalk of IGF and AKT Pathways.

BACKGROUND: Inhibitor of differentiation or DNA binding 1 (ID1) is overexpressed in human salivary gland cancer (SGC). The insulin growth factor (IGF) system is an attractive target in cancer control because it is associated with various cancer progressions. MATERIALS AND METHODS: The human SGC cell line HSY with abundant ID1 was used. ID1 knockdown and its effect on the IGF system were investigated. Cell proliferation and invasion, as well as associated protein expression, were analyzed. Phospho-AKT was also evaluated. RESULTS: ID1 knockdown reduced cell proliferation and invasion, while the expression of proteins associated with malignant phenotypes was altered. IGF-II expression was suppressed, suggesting that this system is one of the mechanisms underlying effects of ID1 in SGC cells. c-Myc was up-regulated, whereas p21 and p27 were down-regulated. Moreover, phospho-AKT was reduced in ID1-knockeddown cells. CONCLUSION: ID1 down-regulation induced parallel changes in the IGF and AKT pathways. The crosstalk of these pathways may enhance malignant phenotypes in SGCs.

Objective validity of an implant-retained overdenture with a ball attachment system after marginal mandibulectomy.

Implant-retained overdentures are known to improve oral function, but the clinical impact on patients who have had mandibular resections is still debatable. We have treated 16 patients who had such resections for oral cancer and consequent loss of the alveolar ridge, with overdentures supported by osseointegrated implants and ball attachments. To quantify their functional improvement, we evaluated their maximum bite force and masticatory performance. Their function improved significantly, (from 77.5N - 365N, 371% increase in maximum bite force, p<0.001) and masticatory performance increased (from 2.5 - 7.7, 208%, p<0.0001) after the overdentures had been inserted. While individual changes in maximum bite force showed no significant correlation, those in masticatory performance correlated significantly, which suggests that the subjects with poor masticatory function are likely to benefit from retention of an implant. These results indicate that implant-retained overdentures are an effective way to rehabilitate patients after marginal mandibular resection.

Estrogen Enhances Malignant Phenotypes in Human Salivary Adenoid Cystic Carcinoma Cells.

BACKGROUND/AIM: Adenoid cystic carcinoma (SGC) is a common type of salivary gland cancer (SGC). Surgery is the first treatment choice because chemoradiotherapy is usually not effective. Therefore, new treatment modalities are urgently needed. In this study, it was investigated whether the estrogen axis could be a treatment target or not. MATERIALS AND METHODS: Adenoid cystic carcinoma (ACC) ACCM cells, were used. The specific cell line lacks estrogen receptor (ER). ER was introduced in ACCM cells, and the effect of 17ß-estradiol (E2) was investigated on cell proliferation, cell-cycle distribution, and cell motility. RESULTS: E2 induced cell proliferation, and the S-phase fraction increased in a dose-dependent manner. Cell motility was also up-regulated compared to control cells. CONCLUSION: The estrogen/ER system up-regulated malignant phenotypes in ER-positive ACC, and hormone therapy may have a potential as effective treatment for this malignancy.

Introduction of ID2 Enhances Invasiveness in ID2-null Oral Squamous Cell Carcinoma Cells via the SNAIL Axis.

AIM: Inhibitor of DNA-binding (ID) proteins are negative regulators of basic helix-loop-helix transcription factors that generally stimulate cell proliferation and inhibit differentiation. However, the role of ID2 in cancer progression remains ambiguous. Here, we investigated the function of ID2 in ID2-null oral squamous cell carcinoma (OSCC) cells. MATERIALS AND METHODS: We introduced an ID2 cDNA construct into ID2-null OSCC cells and compared them with empty-vector-transfected cells in terms of cell proliferation, invasion, and activity and expression of matrix metalloproteinase (MMP). RESULTS: ID2 introduction resulted in enhanced malignant phenotypes. The ID2-expressing cells showed increased N-cadherin, vimentin, and E-cadherin expression and epithelial-mesenchymal transition. In addition, cell invasion drastically increased with increased expression and activity of MMP2. Immunoprecipitation revealed a direct interaction between ID2 and zinc finger transcription factor, snail family transcriptional repressor 1 (SNAIL1). CONCLUSION: ID2 expression triggered a malignant phenotype, especially of invasive properties, through the ID2-SNAIL axis. Thus, ID2 represents a potential therapeutic target for OSCC.

Bite Force and Masticatory Performance Using Implant-supported Overdentures After Treatment of Mandibular Cancer.

AIM: To evaluate overdentures with regard to artificial restoration of oral function following mandibular cancer. MATERIALS AND METHODS: We examined 32 patients who had undergone mandibular bone resection as treatment for malignancy and were using implant-supported overdentures. The patients were aged 55-87 years (mean=68.6) with a male to female ratio of 23:9. Marginal resection was performed in 29 patients and segmentectomy in 3. RESULTS: Before and after using the attachment for overdenture, oral function differed significantly. After the setting of implant-retained overdentures, maximum bite force increased on average by 362% (average, from 16.2 N to 58.8 N; p<0.01). Xylitol gum examination showed a 363% increase in masticatory performance (average, 3.1 to 8.0 points; p<0.01). CONCLUSION: Implant-retained overdenture resulted in improved oral function, that was lost after treatment for mandibular cancer.

Suppression of invasion and metastasis in aggressive salivary cancer cells through targeted inhibition of ID1 gene expression.

Salivary gland cancer (SGC) represents the most common malignancy in the head and neck region, and often metastasizes to the lungs. The helix-loop-helix ID1 protein has been shown to control metastatic progression in many types of cancers. Using two different approaches to target the expression of ID1 (genetic knockdown and progesterone receptor introduction combined with progesterone treatment), we previously determined that the aggressiveness of salivary gland tumor ACCM cells in culture was suppressed. Here, using the same approaches to target ID1 expression, we investigated the ability of ACCM cells to generate lung metastatic foci in nude mice. Moreover, since both approaches would be challenging for applications in humans, we added a third approach, i.e., treatment of mice with a non-toxic cannabinoid compound known to down-regulate ID1 gene expression. All approaches aimed at targeting the pro-metastatic ID1 gene led to a significant reduction in the formation of lung metastatic foci. Therefore, targeting a key transcriptional regulator using different means results in the same reduction of the metastatic spread of SGC cells in animal models, suggesting a novel approach for the treatment of patients with aggressive SGC.

Analysis of isoflavone content in tempeh, a fermented soybean, and preparation of a new isoflavone-enriched tempeh.

To produce a tempeh-like functional food containing a high level of isoflavone with a high absorptivity, we analyzed changes in the composition of isoflavone during tempeh fermentation and the difference in isoflavone content depending on the soybean variety and particular tissue. By adding soybean germ (hypocotyl) that contained a large amount of isoflavone, we prepared a new isoflavone-enriched tempeh in the form of a granular fermented soybean-based food, which can serve as a nutritious supplement for the elderly.

Severe chronic pancreatitis and severe liver cirrhosis have different frequencies and are independent risk factors in male Japanese alcoholics.

BACKGROUND: Chronic pancreatitis and liver cirrhosis are major alcohol-related diseases in most countries. Neither their specific etiologies nor the relationship between them is fully understood. This study was designed to examine a possible association between alcoholic chronic pancreatitis (ACP) and alcoholic liver cirrhosis (ALC), and to identify factors relating to them. METHODS: The subjects were 141 consenting participants from 1087 male patients consecutively admitted to Kurihama National Hospital from July 2000 to November 2002. All were negative for major medical disorders (e.g., viral hepatitis, operative history, malignancy), except for ACP and ALC. Analysis of each subject included background information (collected by face-to-face interview, regarding quantity and duration of drinking, usual alcoholic drinks, smoking, education, employment, and marital status) and signs of ACP detected on endoscopic retrograde cholangiopancreatography (ERCP) and ALC indicated by Child-Pugh classification. Subjects consenting to genome analyses ( n = 83) were genotyped for two key alcohol-metabolizing enzymes: alcohol dehydrogenase-2 and aldehyde dehydrogenase-2. RESULTS: Grouping patients by ERCP grading and Child classification revealed a nonparallel relationship between the severities of the two diseases. This relationship held, even after controlling for several pertinent background variables (sociofamilial, drinking, clinical, and genetic factors) by logistic regression analysis. The drinking of spirits and a high daily consumption of alcohol were independent risk factors for ACP, while never-married status was the only risk factor identified with ALC among these male Japanese patients. CONCLUSIONS: Different risk factors may confer susceptibility to ACP versus ALC, which may explain the nonparallel relationship between the severities of the two diseases in Japanese alcoholics.