An easy-to-implement, non-invasive head restraint method for monkey fMRI.

Functional magnetic resonance imaging (fMRI) in behaving monkeys has a strong potential to bridge the gap between human neuroimaging and primate neurophysiology. In monkey fMRI, to restrain head movements, researchers usually surgically implant a plastic head-post on the skull. Although time-proven to be effective, this technique could create burdens for animals, including a risk of infection and discomfort. Furthermore, the presence of extraneous objects on the skull, such as bone screws and dental cement, adversely affects signals near the cortical surface. These side effects are undesirable in terms of both the practical aspect of efficient data collection and the spirit of "refinement" from the 3R's. Here, we demonstrate that a completely non-invasive fMRI scan in awake monkeys is possible by using a plastic head mask made to fit the skull of individual animals. In all of the three monkeys tested, longitudinal, quantitative assessment of head movements showed that the plastic mask has effectively suppressed head movements, and we were able to obtain reliable retinotopic BOLD signals in a standard retinotopic mapping task. The present, easy-to-make plastic mask has a strong potential to simplify fMRI experiments in awake monkeys, while giving data that is as good as or even better quality than that obtained with the conventional head-post method.

Tadalafil 5 mg Once Daily Improved Each IPSS Subscore, QOL, and Nocturia in Elderly BPH Patients over 70 Years Old in a Real-World Clinical Setting.

OBJECTIVE: No reports have evaluated the treatment effects of tadalafil by age group in a positive, noninterventional observational study of Japanese men. The present study aimed to evaluate the treatment effects of tadalafil by age group in a positive, noninterventional observational study of Japanese men. We therefore divided patients into 2 groups about the age of 70 years and investigated the treatment effects of tadalafil regarding voiding and storage functions by age group. METHODS: Changes from baseline in each parameter (International Prostate Symptom Score [IPSS], quality of life [QOL] score, Overactive Bladder Symptom Score [OABSS], and residual urine volume) at 4, 12, and 24 weeks after initiating tadalafil for benign prostatic hyperplasia (BPH) patients were compared between groups (50-69 years vs. ≥70 years). In addition, side effects of tadalafil were investigated by age group. RESULTS: In the 50-69 years group, significant improvements from baseline were seen in IPSS total and QOL score for all time points. In addition, significant improvements in IPSS storage subscore from baseline were observed at the 4- and 24-week time points. In the ≥70 years group, significant improvements from baseline were seen in IPSS total, IPSS voiding and storage subscores, and QOL score at each time point. CONCLUSIONS: Tadalafil 5 mg once daily appeared effective in clinical settings for elderly BPH patients even over 70 years old.

Multiple neuronal circuits for variable object-action choices based on short- and long-term memories.

At each time in our life, we choose one or few behaviors, while suppressing many other behaviors. This is the basic mechanism in the basal ganglia, which is done by tonic inhibition and selective disinhibition. Dysfunctions of the basal ganglia then cause 2 types of disorders (difficulty in initiating necessary actions and difficulty in suppressing unnecessary actions) that occur in Parkinson's disease. The basal ganglia generate such opposite outcomes through parallel circuits: The direct pathway for initiation and indirect pathway for suppression. Importantly, the direct pathway processes good information and the indirect pathway processes bad information, which enables the choice of good behavior and the rejection of bad behavior. This is mainly enabled by dopaminergic inputs to these circuits. However, the value judgment is complex because the world is complex. Sometimes, the value must be based on recent events, thus is based on short-term memories. Or, the value must be based on historical events, thus is based on long-term memories. Such memory-based value judgment is generated by another parallel circuit originating from the caudate head and caudate tail. These circuit-information mechanisms allow other brain areas (e.g., prefrontal cortex) to contribute to decisions by sending information to these basal ganglia circuits. Moreover, the basal ganglia mechanisms (i.e., what to choose) are associated with cerebellum mechanisms (i.e., when to choose). Overall, multiple levels of parallel circuits in and around the basal ganglia are essential for coordinated behaviors. Understanding these circuits is useful for creating clinical treatments of disorders resulting from the failure of these circuits.

Identification of neuron-type specific promoters in monkey genome and their functional validation in mice.

Viral gene delivery is one of the most versatile techniques for elucidating the mechanisms underlying brain dysfunction, such as neuropsychiatric disorders. Due to the complexity of the brain, expression of genetic tools, such as channelrhodopsin and calcium sensors, often has to be restricted to a specified cell type within a circuit implicated in these disorders. Only a handful of promoters targeting neuronal subtypes are currently used for viral gene delivery. Here, we isolated conserved promoter regions of several subtype-specific genes from the macaque genome and investigated their functionality in the mouse brain when used within lentiviral vectors (LVVs). Immunohistochemical analysis revealed that transgene expression induced by the promoter sequences for somatostatin (SST), cholecystokinin (CCK), parvalbumin (PV), serotonin transporter (SERT), vesicular acetylcholine transporter (vAChT), substance P (SP) and proenkephalin (PENK) was largely colocalized with specific markers for the targeted neuronal populations. Moreover, by combining these results with in silico predictions of transcription factor binding to the isolated sequences, we identified transcription factors possibly underlying cell-type specificity. These findings lay a foundation for the expansion of the current toolbox of promoters suitable for elucidating these neuronal phenotypes.

Direct and indirect pathways for choosing objects and actions.

A prominent target of the basal ganglia is the superior colliculus (SC) which controls gaze orientation (saccadic eye movement in primates) to an important object. This 'object choice' is crucial for choosing an action on the object. SC is innervated by the substantia nigra pars reticulata (SNr) which is controlled mainly by the caudate nucleus (CD). This CD-SNr-SC circuit is sensitive to the values of individual objects and facilitates saccades to good objects. The object values are processed differently in two parallel circuits: flexibly by the caudate head (CDh) and stably by the caudate tail (CDt). To choose good objects, we need to reject bad objects. In fact, these contrasting functions are accomplished by the circuit originating from CDt: The direct pathway focuses on good objects and facilitates saccades to them; the indirect pathway focuses on bad objects and suppresses saccades to them. Inactivation of CDt deteriorated the object choice, because saccades to bad objects were no longer suppressed. This suggests that the indirect pathway is important for object choice. However, the direct and indirect pathways for 'object choice', which aim at the same action (i.e., saccade), may not work for 'action choice'. One possibility is that circuits controlling different actions are connected through the indirect pathway. Additional connections of the indirect pathway with brain areas outside the basal ganglia may also provide a wider range of behavioral choice. In conclusion, basal ganglia circuits are composed of the basic direct/indirect pathways and additional connections and thus have acquired multiple functions.

Medial thalamus in the territory of oculomotor basal ganglia represents stable object value.

Many visual objects are attached with values which were created by our long rewarding history. Such stable object values attract gaze. We previously found that the output pathway of basal ganglia from caudal-dorsal-lateral portion of substantia nigra pars reticulata (cdlSNr) to superior colliculus (SC) carries robust stable value signal to execute the automatic choice of valuable objects. An important question here is whether stable value signal in basal ganglia can influence on other inner processing such as perception, attention, emotion, or arousal than motor execution. The key brain circuit is another output path of basal ganglia: the pathway from SNr to temporal and frontal lobes through thalamus. To examine the existence of stable value signal in this pathway, we explored thalamus in a wide range. We found that many neurons in the medial thalamus represented stable value. Histological examination showed that the recorded sites of those neurons included ventral anterior nucleus, pars magnocellularis (VAmc) which is the main target of nigrothalamic projection. Consistent with the SNr GABArgic projection, the latency of value signal in the medial thalamus was later than cdlSNr, and the sign of value coding in the medial thalamus was opposite to cdlSNr. As is the case with cdlSNr neurons, the medial thalamus neurons showed no sensitivity to frequently updated value (flexible value). These results suggest that the pathway from cdlSNr to the medial thalamus influences on various aspects of cognitive processing by propagating stable value signal to the wide cortical area.

To Wait or Not to Wait-Separate Mechanisms in the Oculomotor Circuit of Basal Ganglia.

We reach a goal immediately after detecting the target, or later by withholding the immediate action. Each time, we choose one of these actions by suppressing the other. How does the brain control these antagonistic actions? We hypothesized that the output of basal ganglia (BG), substantia nigra pars reticulata (SNr), suppresses antagonistic oculomotor signals by sending strong inhibitory output to superior colliculus (SC). To test this hypothesis, we trained monkeys to perform two kinds of saccade task: Immediate (visually guided) and delayed (visually-withheld but memory-guided) saccade tasks. In both tasks, we applied one-direction-reward (1DR) procedure to modify the level of goal-reaching motivation. We identified SNr neurons that projected to SC by their antidromic activation from SC. We stimulated SC on both sides because SNr neurons projecting to the ipsilateral SC (ipsiSC) and those projecting to the contralateral SC (contraSC) might have antagonistic functions. First, we found that ipsiSC-projecting neurons were about 10 times more than contraSC-projecting neurons. More importantly, ipsiSC-projecting SNr neurons were roughly divided into two groups which would control immediate and delayed saccades separately. The immediate-type SNr neurons were clearly inhibited by a visual target on the contralateral side in both visual- and memory-1DR tasks. The inhibition would disinhibit SC neurons and facilitate a saccade to the contralateral target. This is goal-directed in visual-1DR task, but is erroneous in memory-1DR task. In contrast, the delayed-type SNr neurons tended to be excited by a visual target (especially on the contralateral side), which would suppress the immediate saccade to the target. Instead, they were inhibited before a delayed (memory-guided) saccade directed to the contralateral side, which would facilitate the saccade. ContraSC-projecting SNr neurons were more variable with no grouped features, although some of them may contribute to the saccade to the ipsilateral target. Finally, we found that some ipsiSC-projecting SNr neurons were inhibited more strongly when reward was expected, which was associated with shortened saccade reaction times. However, many SNr neurons showed no reward-expectation effect. These results suggest that two separate oculomotor circuits exist in BG, both of which contribute to goal-directed behavior, but in different temporal contexts.

Functional territories in primate substantia nigra pars reticulata separately signaling stable and flexible values.

Gaze is strongly attracted to visual objects that have been associated with rewards. Key to this function is a basal ganglia circuit originating from the caudate nucleus (CD), mediated by the substantia nigra pars reticulata (SNr), and aiming at the superior colliculus (SC). Notably, subregions of CD encode values of visual objects differently: stably by CD tail [CD(T)] vs. flexibly by CD head [CD(H)]. Are the stable and flexible value signals processed separately throughout the CD-SNr-SC circuit? To answer this question, we identified SNr neurons by their inputs from CD and outputs to SC and examined their sensitivity to object values. The direct input from CD was identified by SNr neuron's inhibitory response to electrical stimulation of CD. We found that SNr neurons were separated into two groups: 1) neurons inhibited by CD(T) stimulation, located in the caudal-dorsal-lateral SNr (cdlSNr), and 2) neurons inhibited by CD(H) stimulation, located in the rostral-ventral-medial SNr (rvmSNr). Most of CD(T)-recipient SNr neurons encoded stable values, whereas CD(H)-recipient SNr neurons tended to encode flexible values. The output to SC was identified by SNr neuron's antidromic response to SC stimulation. Among the antidromically activated neurons, many encoded only stable values, while some encoded only flexible values. These results suggest that CD(T)-cdlSNr-SC circuit and CD(H)-rvmSNr-SC circuit transmit stable and flexible value signals, largely separately, to SC. The speed of signal transmission was faster through CD(T)-cdlSNr-SC circuit than through CD(H)-rvmSNr-SC circuit, which may reflect automatic and controlled gaze orienting guided by these circuits.

Effects of luminance contrast on the color selectivity of neurons in the macaque area v4 and inferior temporal cortex.

Appearance of a color stimulus is significantly affected by the contrast between its luminance and the luminance of the background. In the present study, we used stimuli evenly distributed on the CIE-xy chromaticity diagram to examine how luminance contrast affects neural representation of color in V4 and the anterior inferior temporal (AITC) and posterior inferior temporal (PITC) color areas (Banno et al., 2011). The activities of single neurons were recorded from monkeys performing a visual fixation task, and the effects of luminance contrast on the color selectivity of individual neurons and their population responses were systematically examined by comparing responses to color stimuli that were brighter or darker than the background. We found that the effects of luminance contrast differed considerably across V4 and the PITC and AITC. In both V4 and the PITC, the effects of luminance contrast on the population responses of color-selective neurons depended on color. In V4, the size of the effect was largest for blue and cyan, whereas in the PITC, the effect gradually increased as the saturation of the color stimulus was reduced, and was especially large with neutral colors (white, gray, black). The pattern observed in the PITC resembles the effect of luminance contrast on color appearance, suggesting PITC neurons are closely involved in the formation of the perceived appearance of color. By contrast, the color selectivities of AITC neurons were little affected by luminance contrast, indicating that hue and saturation of color stimuli are represented independently of luminance contrast in the AITC.

Basal ganglia circuits for reward value-guided behavior.

The basal ganglia are equipped with inhibitory and disinhibitory mechanisms that enable a subject to choose valuable objects and actions. Notably, a value can be determined flexibly by recent experience or stably by prolonged experience. Recent studies have revealed that the head and tail of the caudate nucleus selectively and differentially process flexible and stable values of visual objects. These signals are sent to the superior colliculus through different parts of the substantia nigra so that the animal looks preferentially at high-valued objects, but in different manners. Thus, relying on short-term value memories, the caudate head circuit allows the subject's gaze to move expectantly to recently valued objects. Relying on long-term value memories, the caudate tail circuit allows the subject's gaze to move automatically to previously valued objects. The basal ganglia also contain an equivalent parallel mechanism for action values. Such flexible-stable parallel mechanisms for object and action values create a highly adaptable system for decision making.

Why skill matters.

Maximizing rewards per unit time is ideal for success and survival in humans and animals. This goal can be approached by speeding up behavior aiming at rewards and this is done most efficiently by acquiring skills. Importantly, reward-directed skills consist of two components: finding a good object (i.e., object skill) and acting on the object (i.e., action skill), which occur sequentially. Recent studies suggest that object skill is based on high-capacity memory for object-value associations. When a learned object is encountered the corresponding memory is quickly expressed as a value-based gaze bias, leading to the automatic acquisition or avoidance of the object. Object skill thus plays a crucial role in increasing rewards per unit time.

Robust representation of stable object values in the oculomotor Basal Ganglia.

Our gaze tends to be directed to objects previously associated with rewards. Such object values change flexibly or remain stable. Here we present evidence that the monkey substantia nigra pars reticulata (SNr) in the basal ganglia represents stable, rather than flexible, object values. After across-day learning of object-reward association, SNr neurons gradually showed a response bias to surprisingly many visual objects: inhibition to high-valued objects and excitation to low-valued objects. Many of these neurons were shown to project to the ipsilateral superior colliculus. This neuronal bias remained intact even after >100 d without further learning. In parallel with the neuronal bias, the monkeys tended to look at high-valued objects. The neuronal and behavioral biases were present even if no value was associated during testing. These results suggest that SNr neurons bias the gaze toward objects that were consistently associated with high values in one's history.

What and where information in the caudate tail guides saccades to visual objects.

We understand the world by making saccadic eye movements to various objects. However, it is unclear how a saccade can be aimed at a particular object, because two kinds of visual information, what the object is and where it is, are processed separately in the dorsal and ventral visual cortical pathways. Here, we provide evidence suggesting that a basal ganglia circuit through the tail of the monkey caudate nucleus (CDt) guides such object-directed saccades. First, many CDt neurons responded to visual objects depending on where and what the objects were. Second, electrical stimulation in the CDt induced saccades whose directions matched the preferred directions of neurons at the stimulation site. Third, many CDt neurons increased their activity before saccades directed to the preferred objects and directions of the neurons in a free-viewing condition. Our results suggest that CDt neurons receive both "what" and "where" information and guide saccades to visual objects.

SNRPE is involved in cell proliferation and progression of high-grade prostate cancer through the regulation of androgen receptor expression.

Clinically high-grade prostate cancers (PC) with high Gleason scores of 8-10 exhibit rapid growth and are more likely to spread beyond the prostate. These cancer types demonstrate a poor response to androgen deprivation therapy and eventually acquire a castration-resistant phenotype. To identify novel molecular cancer drug targets, we previously analyzed the gene expression profiles of high-grade PC using a cDNA microarray combined with laser microbeam microdissection and found a number of genes that are transactivated in high-grade PC. Among these genes, we report the identification of a novel molecular target, small nuclear ribonucleoprotein polypeptide E (SNRPE). Semi-quantitative RT-PCR confirmed that SNRPE is overexpressed in high-grade PC cells compared with normal prostatic epithelial cells. Knockdown of SNRPE expression by short interfering RNA (siRNA) resulted in the marked suppression of PC cell proliferation. By contrast, SNRPE overexpression promoted PC cell proliferation, indicating its oncogenic effects. Furthermore, we demonstrated that SNRPE regulates androgen receptor (AR) mRNA expression in PC cells. Knockdown of SNRPE expression by siRNA resulted in the marked suppression of AR and its downstream target genes at the mRNA level. We suggest that the regulation of AR expression by SNRPE is essential for cell proliferation and progression of high-grade PC and that it may be a novel molecular target for cancer drugs.

Phenolic constituents isolated from Fragaria ananassa Duch. inhibit antigen-stimulated degranulation through direct inhibition of spleen tyrosine kinase activation.

We isolated eight phenolic constituents from Fragaria ananassa Duch. (strawberry) and determined their structures using 1D, 2D-NMR. Among the isolated compounds, linocinnamarin (LN), 1-O-trans-cinnamoyl-beta-d-glucopyranose (CG), and cinnamic acid (CA) exhibited antigen (Ag)-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells. In order to reveal the underlying mechanisms, we examined the effects of LN and CA on cellular responses induced by antigen stimulation. Treatment with both LN and CA markedly inhibited antigen-stimulated elevation of intracellular free Ca(2+) concentration and reactive oxygen species (ROS). Both LN and CA suppressed Ag-stimulated spleen tyrosine kinase (Syk) activation. These results indicate that inhibition of antigen-stimulated degranulation by LN and CA is mainly due to inactivation of Syk/phospholipase Cgamma (PLCgamma) pathways. Our findings suggest that LN and CA isolated from F. ananassa Duch. (strawberry) could be beneficial agents for alleviating symptoms of type I allergy.

Pseudosarcomatous proliferation of cx43- and kit-expressing interstitial cell in the urinary bladder.

The authors report a case showing proliferation of KIT- and connexin 43-expressing mesenchymal cells of the urinary bladder. A 75-year-old woman had an ulcerated endophytic mass (size, approximately 2 × 2 cm) in the left posterolateral wall. She underwent transurethral resection and subsequent partial cystectomy. The suburothelial mass extended to the muscularis propria. The histopathological analysis revealed spindle-shaped mesenchymal cells that were loosely arranged with myxoid stroma and showed a focal compact fascicular arrangement. In the immunohistochemical analysis, these spindle cells were stained with specific antibodies to KIT and connexin 43. The patient is currently free of disease at 5 years after operation. The proliferating spindle cells in the present case might represent a phenotype of interstitial cells of the lamina propria.

Color selectivity of neurons in the posterior inferior temporal cortex of the macaque monkey.

We recorded the activities of neurons in the lateral surface of the posterior inferior temporal cortex (PIT) of 3 hemispheres of 3 monkeys performing a visual fixation task. We characterized the color and shape selectivities of each neuron, mapped its receptive field (RF), and studied the distributions of these response properties. Using a set of color stimuli that were systematically distributed in Commission Internationale de l'Eclairage-xy chromaticity diagram, we found numerous color-selective neurons distributed throughout the area examined. Neurons in the ventral region tended to have sharper color tuning than those in the dorsal region. We also found a crude retinotopic organization in the ventral region. Within the ventral region of PIT, neurons in the dorsal part had RFs that overlapped the foveal center; the eccentricity of RFs increased in the more ventral part, and neurons in the anterior and posterior parts had RFs that represented the lower and upper visual fields, respectively. In all 3 hemispheres, the region where sharply tuned color-selective neurons were concentrated was confined within this retinotopic map. These findings suggest that PIT is a heterogeneous area and that there is a circumscribed region within it that has crude retinotopic organization and is involved in the processing of color.

[Clinical experience with transurethral resection of bladder tumor (TUR-Bt) guided by photodynamic diagnosis (PDD)].

PURPOSE: To report our clinical experience regarding transurethral resection of bladder tumor (TUR-Bt) guided by photodynamic diagnosis (PDD) with intravesical instillations of 5-aminolevulinic acid (ALA) and to assess the usefulness of the therapeutic method. MATERIALS AND METHODS: TUR-Bt guided by PDD was performed in 57 patients of which 47 were men and 10 women with a median age of 74.3 years (range 45-90), 36 were primary cases and 21 were recurrent cases with non-muscle invasive bladder cancer. Two to two and half hours prior to endoscopy 1.5 g ALA dissolved in 50 ml of 8.4% sodium hydrogen carbonate (NaHCO3) solution was instilled intravesically. For fluorescence excitation a blue light source (D-LIGHT System, Karl Storz Endoscopy Japan K.K.) was used. The tumorous lesions under white light guidance and the lesion with fluorescent excitation under blue (fluorescence) light guidance were taken by cold cup as a biopsy and also resected sequentially. To evaluate the accuracy of PDD, the levels in images of the ALA-induced fluorescence were compared with the pathological results. To evaluate the availability of TUR-Bt guided by PDD, survival Analysis regarding vesical recurrence was retrospectively examined compared to the cases underwent conventional TUR-Bt under white light guidance. Moreover, in these cases, multivariate analysis using Cox proportional-hazards model was performed to detect the clinico-pathological factor independently contribute to improving prognosis. (Results) In the 301 specimens obtained from 57 patients, the sensitivity and specificity of PDD were 92.5% and 60.1%, whereas the sensitivity and specificity of conventional endoscopic examination under white light guidance were 81.6% and 79.5%, respectively. Median follow-up period was 19.1 (range 8.6-49.9) months in 57 patients underwent TUR-Bt guided by PDD. Eight of 57 patients recurred and recurrence-free survival rate was 88.2 +/- 0.1% (at 12 months) and 76.2 +/- 0.1% (24-48 months). Median follow-up period was 49.9 (5.0-145.0) months in 149 patients underwent conventional TUR-Bt. Ninety-nine of 149 patients recurred and recurrence-free survival rate was 60.3 +/- 0.0% (12 months) and 31.6 +/- 0.0% (24-48 months). There was statistical significance in recurrence-free survival rate between these 2 therapeutic groups (p < 0.001). Moreover, multivariate analysis revealed the independent factor contribute to improving prognosis was only TUR-Bt guided by PDD (hazard ratio 0.279, p = 0.001). CONCLUSION: It was suggested that TUR-Bt guided by PDD might reduce the risk of vesical recurrence in the early stage after operation of non-muscle invasive bladder cancer.

[Experience in the treatment of eight cases of Fournier's gangrene].

Fournier's gangrene is a rare disease of rapidly progressive necrotising fasciitis of the genital, perineal and perianal regions and leads to sepsis and death. We report 8 cases of Fournier's gangrene treated at our hospital and affiliated hospitals from 1997 to 2007. There were seven males and one female in the series, and the age range was 23-89 years (mean age 56.6 years). Four patients among them had diabetes mellitus. We rescued all patients by broad-spectrum antibacterial chemotherapy and debridement. Good management should be based on broad-spectrum antibacterial chemotherapy, debridement and intensive care.

Inhibitory effects of flavonoids isolated from Fragaria ananassa Duch on IgE-mediated degranulation in rat basophilic leukemia RBL-2H3.

We isolated the 4 kinds of flavonoids from strawberry 'Nohime' and examined the effect of these flavonoids on the degranulation in RBL-2H3 cells. The flavonoids were found to suppress the degranulation from Ag-stimulated RBL-2H3 cells to different extents. To disclose the inhibitory mechanism of degranulation by flavonoids, we examined their effects on the intracellular free Ca(2+) concentration ([Ca(2+)]i) and the intracellular signaling pathway such as Lyn, Syk, and PLCgammas. The intracellular free Ca(2+) concentration ([Ca(2+)]i) was elevated by Fc epsilonRI activation, but these flavonoid treatments reduced the elevation of [Ca(2+)]i by suppressing Ca(2+) influx. Kaempferol strongly suppressed the activation of Syk and PLCgammas. It was thus suggested that suppression of Ag-stimulated degranulation by the flavonoids is mainly due to suppression of [Ca(2+)]i elevation and Syk activation. These results suggested that strawberry would be of some ameliorative benefit for the allergic symptoms.