Sensitization of TRPV1 by protein kinase C in rats with mono-iodoacetate-induced joint pain.

OBJECTIVE: To assess the functional changes of Transient receptor potential vanilloid 1 (TRPV1) receptor and to clarify its mechanism in a rat mono-iodoacetate (MIA)-induced joint pain model (MIA rats), which has joint degeneration with cartilage loss similar to osteoarthritis. METHODS: Sensitization of TRPV1 in MIA rats was assessed by transient spontaneous pain behavior induced by capsaicin injection in knee joints and electrophysiological changes of dorsal root ganglion (DRG) neurons innervating knee joints in response to capsaicin. Mechanisms of TRPV1 sensitization were analyzed by a newly developed sandwich enzyme-linked immunosorbent assay that detects phosphorylated TRPV1, followed by functional and expression analyses of protein kinase C (PKC) in vivo and in vitro, which involves TRPV1 phosphorylation. RESULTS: Pain-related behavior induced by intra-articular injection of capsaicin was significantly increased in MIA rats compared with sham rats. In addition, capsaicin sensitivity, evaluated by capsaicin-induced inward currents, was significantly increased in DRG neurons of MIA rats. Protein levels of TRPV1 remained unchanged, but phosphorylated TRPV1 at Ser800 increased in DRG neurons of MIA rats. Phosphorylated-PKCɛ (p-PKCɛ) increased and co-localized with TRPV1 in DRG neurons of MIA rats. Capsaicin-induced pain-related behavior in MIA rats was inhibited by intra-articular pretreatment of the PKC inhibitor bisindolylmaleimide I. In addition, intra-articular injection of the PKC activator phorbol 12-myristate 13-acetate increased capsaicin-induced pain-related behavior in normal rats. CONCLUSION: TRPV1 was sensitized at the knee joint and at DRG neurons of MIA rats through PKC activation. Thus, TRPV1 sensitization might be involved in chronic pain caused by osteoarthritis.

An acquired distaste: sugar discrimination by the larval parasitoid Microplitis croceipes (Hymenoptera: Braconidae) is affected by prior sugar exposure.

We examined gustatory responses of the larval parasitoid Microplitis croceipes to determine whether the adults discriminate among common sugars, including fructose, glucose, maltose and sucrose, found in plants. When given single sugar solutions of sucrose, glucose, fructose and maltose at concentrations of 0.008-2.0 mol l(-1), the estimated concentrations at which 50% of wasps initiated feeding ranged between 0.054 and 0.085 mol l(-1) for sucrose, glucose and fructose, which was significantly lower than for maltose. Wasps showed a strong decrease in feeding time for maltose or fructose following a brief exposure to other sugars, suggesting that wasps can distinguish maltose and fructose from the other sugars tested. The higher acceptance threshold and short feeding time in the case of maltose appears adaptive in light of the relatively poor nutritional quality of the sugar in the longevity trial. The pronounced feeding inhibition seen for fructose following exposure to other sugars is not linked with lower nutritional performance. This feeding inhibition was even seen in wasps that had fed on glucose at the lowest acceptance threshold (0.031 mol l(-1)) and persisted for 24 h. This study is the first to show feeding inhibition of otherwise phagostimulant sugars such as maltose and fructose after gustatory stimulation on other sugars.

Pharmacological characterization of lysophosphatidic acid-induced pain with clinically relevant neuropathic pain drugs.

Lysophosphatidic acid (LPA), an initiator of neuropathic pain, causes allodynia. However, few studies have evaluated the pharmacological profile of LPA-induced pain. In this study, a LPA-induced pain model was developed and pharmacologically characterized with clinically relevant drugs used for neuropathic pain, including antiepileptics, non-steroidal anti-inflammatory agents, analgesics, local anaesthetics/antiarrhythmics and antidepressants. Gabapentin (1-30 mg/kg, p.o.) significantly reversed LPA-induced allodynia, but neither indomethacin (30 mg/kg, p.o.) nor morphine (0.3-3 mg/kg, s.c.) did, which indicates that LPA-induced pain consists mostly of neuropathic rather than inflammatory pain. Both pregabalin (0.3-10 mg/kg, p.o.) and ω-CgTX MVIIA (0.01-0.03 µg/mouse, i.t.) completely reversed LPA-induced allodynia in a dose-dependent manner. Lidocaine (1-30 mg/kg, s.c.), mexiletine (1-30 mg/kg, p.o.) and carbamazepine (10-100 mg/kg, p.o.) significantly ameliorated LPA-induced allodynia dose dependently. Milnacipran (30 mg/kg, i.p.) produced no significant analgesic effect in LPA-induced allodynia. In LPA-injected mice, expression of the α2δ1 subunit of the voltage-gated calcium channel (VGCC) was increased in the dorsal root ganglion (DRG) and spinal dorsal horn. Furthermore, the VGCC current was potentiated in both the DRG from LPA-injected mice and LPA (1 µM)-treated DRG from saline-injected mice, and the potentiated VGCC current was amended by treatment with gabapentin (100 µM). The LPA-induced pain model described here mimics aspects of the neuropathic pain state, including the sensitization of VGCC, and may be useful for the early assessment of drug candidates to treat neuropathic pain.

The synthetic TRH analogue taltirelin exerts modality-specific antinociceptive effects via distinct descending monoaminergic systems.

BACKGROUND AND PURPOSE: Exogenously administered thyrotropin-releasing hormone (TRH) is known to exert potent but short-acting centrally-mediated antinociceptive effects. We sought to investigate the mechanisms underlying these effects using the synthetic TRH analogue taltirelin, focusing on the descending monoaminergic systems in mice. EXPERIMENTAL APPROACH: The mice received systemic or local injections of taltirelin combined with either central noradrenaline (NA) or 5-hydroxytryptamine (5-HT) depletion by 6-hydroxydopamine (6-OHDA) or DL-p-chlorophenylalanine (PCPA), respectively, or blockade of their receptors. The degree of antinociception was determined using the tail flick and tail pressure tests. KEY RESULTS: Subcutaneously (s.c.) administered taltirelin exhibited dose-dependent antinociceptive effects in the tail flick and tail pressure tests. These effects appeared to be primarily supraspinally mediated, since intracerebroventricularly (i.c.v.) but not intrathecally (i.t.) injected taltirelin generated similar effects. Depletion of central NA abolished only the analgesic effect of taltirelin (s.c. and i.c.v.) on mechanical nociception. By contrast, depletion of central 5-HT abolished only its analgesic effect on thermal nociception. Intraperitoneal (i.p.) and i.t. injection of the alpha2-adrenoceptor antagonist yohimbine respectively reduced the analgesic effect of taltirelin (s.c. and i.c.v.) on mechanical nociception. By contrast, the 5-HT1A receptor antagonist WAY-100635 (i.p. and i.t.) reduced the effect of taltirelin (s.c. and i.c.v.) on thermal nociception. Neither the 5-HT2 receptor antagonist ketanserin nor the opioid receptor antagonist naloxone altered the antinociceptive effect of taltirelin. CONCLUSIONS AND IMPLICATIONS: These findings suggest that taltirelin activates the descending noradrenergic and serotonergic pain inhibitory systems, respectively, to exert its analgesic effects on mechanical and thermal nociception.

Parasitic wasps learn and report diverse chemicals with unique conditionable behaviors.

Parasitoids exploit numerous chemical cues to locate hosts and food. Whether they detect and learn chemicals foreign to their natural history has not been explored. We show that the parasitoid Microplitis croceipes can associate, with food or hosts, widely different chemicals outside their natural foraging encounters. When learned chemicals are subsequently detected, this parasitoid manifests distinct behaviors characteristic with expectations of food or host, commensurate with prior training. This flexibility of parasitoids to rapidly link diverse chemicals to resource needs and subsequently report them with recognizable behaviors offers new insights into their foraging adaptability, and provides a model for further dissection of olfactory learning related processes.

Facile and stereoselective access to nonracemic tricyclic cyclobutanes by asymmetric intramolecular Michael-aldol reaction: thermodynamic equilibrium and activation by iodonium ion.

Intramolecular Michael-aldol reactions of (-)-phenylmenthyl enoates tethered to cycloalkanone affords tricyclic cyclobutanes with high degrees of diastereochemical control. Kinetic and thermodynamic studies revealed that the Michael-aldol reaction is reversible under conditions in which trimethylsilyl iodide is used in the presence of hexamethyldisilazane at ambient temperature. Different levels of diastereoselectivity are observed when this cyclization process is carried out under kinetic vs thermodynamic conditions. Finally, an influence of added iodonium donors on the reaction rate has been noted.

Conformation of 1,4-dineopentyl-2,5-cis-diphenylpiperazine and its diammonium salts: remarkable change in conformation depending upon the counter anion.

The free base of 1,4-dineopentyl-2,5-diphenylpiperazine takes a chair conformation in CDCl3, while the conformation of its diammonium salts changes depending upon the counter anion.

[Development of boomerang-type intramolecular cascade reactions and application to natural product synthesis].

Intramolecular cascade reaction has received much attention as a powerful methodology to construct a polycyclic framework in organic synthesis. We have been developing "boomerang-type cascade reaction" to construct a variety of polycyclic skeletons efficiently. In the above reactions, a nucleophilic function of substrates changes the character into an electrophile after the initial reaction, and the electrophilic group acts as a nucleophile in the second reaction. That is, the reaction center stepwise moves from one functional group back to the same one via other functional groups. The stream of the electron concerning the cascade reaction is like a locus of boomerang. We show here three different boomerang-type reactions via ionic species or free radicals. 1) Diastereoselective Michael-aldol reaction based on the chiral auxiliary method and enantioselective Michael-aldol reaction by the use of external chiral sources. 2) Short and efficient total syntheses of longifolane sesquiterpenes utilizing intramolecular double Michael addition as a key step. 3) Development of boomerang-type radical cascade reaction of halopolyenes to construct terpenoid skeletons and its regioselectivity.

Novel intramolecular (4 + 1) and (4 + 2) annulations of halopolyenes by cascade radical reaction

Free radical reaction of vinyl iodides having dinenoate function in the presence of tributyltin hydride or tris(trimethylsilyl)silane caused a sequential cyclization reaction to produce (4 + 1) and (4 + 2) annulated compounds by means of a cascade radical reaction. Stereogenic centers of the cascade reaction were highly controlled. On the contrary, the cathodic electrolysis of the vinyl iodides afforded monocyclic compounds.

Primary malignant melanoma of the male urethra.

BACKGROUND: We describe a rare case of malignant melanoma originating from the male urethra. METHODS/RESULTS: This case, which presented a tan-colored lesion at the distal urethra on urethroscopy, was considered as transitional cell carcinoma by transurethral biopsy. Pathologic examination of widely resected urethra revealed scattering cells, which included melanin granules in cytoplasm, and then immunohistochemical stains established the diagnosis as amelanotic melanoma. Although partial penectomy and inguinal lymph node dissection were subsequently performed, 7 months later, total cystourethrectomy was required because of multifocal recurrence at the residual urethra. CONCLUSIONS: In cases of amelanotic melanoma, it should be noted that a lack of melanin granules in tumor cells may lead pathologists and urologists to a misdiagnosis due to the histologic non-specific appearance. With respect to surgical management, of significance is the fact that malignant melanoma may progress multifocally along the total urethra; therefore, total urethrectomy should be considered as radical surgery for the primary site, even if the tumor is confined to the distal urethra.

[Papillary adenocarcinoma of the prostate: report of 3 cases].

We report here 3 cases of papillary adenocarcinoma of the prostate. In all 3 cases, the tumors were discernible on cystourethroscopy and transurethral biopsy established the diagnosis, whereas no significant finding was found on digital rectal examination. Although androgen deprivation therapy was administered in all cases, different surgical procedures were employed according to the stage in each case. In case 1, since the papillary tumor was confined within the prostatic urethra, complete resection was accomplished by transurethral resection (TUR). In case 2, since pelvic lymph nodes metastases were found, local radiation therapy was added. In case 3, since the patient had vesical invasion of tumor total cysto-prostatectomy was performed. Papillary adenocarcinoma of the prostate originates from the prostatic duct, resulting in existence at the "central portion" of the prostate gland. Cystourethroscopy and transurethral biopsy is helpful for diagnosis of this disease, whereas rectal digital examination is useless. As a surgical procedure for the primary site, TUR may be efficient for tumors confined within the prostatic urethra, although more extensive surgery may be necessary for those with a more invasive profile.

Total synthesis of (+/-)-culmorin and (+/-)-longiborneol: an efficient construction of Tricyclo[6.3.0.0(3,9)]undecan-10-one by intramolecular double Michael addition.

The treatment of 4-[(5E)-6-methoxycarbonyl-5-hexenyl]-3, 4-dimethyl-2-cyclopenten-1-one (5) with LHMDS, TMSI-HMDS, Bu(2)OTf-HMDS, or TMSCl-NEt(3)-ZnCl(2) caused the intramolecular double Michael addition to afford tricyclo[6.3.0.0(3, 9)]undecan-10-one 12 in high yields with perfect stereoselectivity. The methodology was further elaborated to achieve efficient total syntheses of (+/-)-culmorin (1) and (+/-)-longiborneol (2). The common precursor 13 of them was obtained from 14 in 94% yield as a single isomer by the treatment with LHMDS. After the conversion of 13 into the corresponding acid 24 by hydrolysis, oxidative decarboxylation using S-(1-oxido-2-pyridinyl)-1,1,3, 3-tetramethylthiouronium hexafluorophosphate (HOTT, 27), followed by the Birch reduction, stereoselectively afforded (+/-)-culmorin (1). (+/-)-Longiborneol (2) was synthesized from 24 by the standard transformation. Additionally, the treatment of 24 with Pb(OAc)(4) led to 28 via uncommon migration. Its structure was determined by X-ray analysis after the transformation into the diketone 29.

Conformational difference between mono- and diprotonated cis-2,5-diphenylpiperazinium salts in the solid state.

Monohydrochlorides of cis-2,5-diphenylpiperazine assume a chair conformation, while the corresponding dihydrochlorides assume a boat form regardless of the substituent(s) at the nitrogen atom.

Case report: glomus tumour of the nasal cavity and paranasal sinuses.

Glomus tumours are relatively rare in the head and neck. We present a glomus tumour of the nasal cavity and paranasal sinuses in a 55-year-old man and describe the CT appearances of this tumour and its histopathology.

Immunohistochemical evidence of the mesothelial histogenesis of a uterine adenomatoid tumor.

We present rare case of a uterine adenomatoid tumor. In order to characterize the diversity of immunoreactive antigens associated with mesothelial differentiation, we immunohistochemically examined the tumor's epithelioid and myofibromatoid components, as well as the biphasic pattern of its adenomatoid changes. Immunostaining of tumor cells was positive for high- and low-molecular-weight cytokeratins and vimentin. Specific immunoreactions with antibodies against desmin, alpha-actin, and the S-100 protein also were observed. The present adenomatoid tumor can be explained in terms of various transformations and typical alterations in mesothelioma cells: (1) the mesothelial cells had differentiated to epithelial and stromal components, concomitant with the presence of predominant reactive foci; (2) intermediate cells expressed different types of cytoskeletal intermediate filament proteins (IMPs); and (3) the patterns of fibromatoid and leiomyoid differentiation resembled those of a benign mesothelioma of the ovarian or oviductal peritoneum. Our immunohistochemical investigations indicated that the present tumor exhibited the histogenesis of a true mesothelioma, an adenomatoid mesothelioma.

[Immunohistochemical study of thymidine phosphorylase in uterine cervical intraepithelial neoplasia].

Studies have shown that thymidine phosphorylase (TdRPase) activity in various tumors is higher than that in normal tissues. We studied the immunohistochemical localization of TdRPase in uterine cervical intraepithelial neoplasia (CIN) using monoclonal antibody against TdRPase. Sixty-nine patients were studied: CIN 1, 14; CIN 2, 15; CIN 3 (severe dysplasia), 23; CIN 3 (carcinoma in situ), 17. Immunoreactivity for TdRPase was found in 29% of CIN 1, 40% of CIN 2, 70% of CIN 3 (severe dysplasia), and 88% of CIN 3 (carcinoma in situ), showing increasing incidence with the progression of tumor grade. Furthermore, in higher grade tumor, more immunoreactive cells were found with increased intensity of immunoreaction. Both nucleus and cytoplasm of atypical cells showed immunoreactivity in most TdRPase positive cases, while either nucleus or cytoplasm was positive in a few cases. The stromal tissues were negative for the antibody except 3 cases of CIN 3 (carcinoma in situ) in which the stromal tissues adjacent to the tumor was positive. Our data suggest that immunoreactivity of TdRPase in CIN may correlate with the grade of CIN.

Various manifestations of early and minimally advanced gastric cancer in gastrointestinal series (GIS).

Preoperative gastrointestinal series (GIS) have been used to assess the precise localization and extent of early and minimally advanced gastric cancers, which are sometimes difficult to define en face by endoscopy. The purpose of this study was to relate various GIS manifestations of gastric cancer with the pathological condition. We reviewed 99 gastric cancer cases (107 lesions), which were treated between 1992 and 1995, in which the lesions were depicted by GIS assisted by a nasogastric tube. The pathological conditions were determined from the resected specimens. Differences in anatomic location and in some morphological characteristics contribute to differences in histological classification. We conclude that lesion depiction by GIS assisted by nasogastric tube reflects the cancer's pathological characteristics, including histological type, malignant cycle, and modification by desmoplastic reaction.