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.

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.

Occurrence of methionine-enkephalin-Arg6-Gly7-Leu8 with methionine-enkephalin, leucine-enkephalin and methionine-enkephalin-Arg6-Phe7 in human gastric antrum.

Recent studies on the nucleotide sequence of cloned DNA complementary to mRNA for preproenkephalin from bovine adrenal medulla and human pheochromocytoma have revealed that this precursor molecule contains four copies of methionine-enkephalin (Met-Enk) and one copy each of leucine-enkephalin (Leu-Enk), methionine-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg6-Gly7-Leu8) and methionine-enkephalin-Arg6-Phe7 (Met-Enk-Arg6-Phe7). We have demonstrated the existence of Met-Enk-Arg6-Gly7-Leu8 together with Met-Enk, Leu-Enk and Met-Enk-Arg6-Phe7 in human gastric antrum, using high performance liquid chromatography (HPLC) coupled with radioimmunoassays for these opioid peptides. The ratio of molar concentrations of these peptides in human gastric antrum is similar to the ratio of these peptides contained in preproenkephalin. Furthermore, gel filtration studies on Sephadex G-50 showed that most of immunoreactivities of these peptides were eluted at the elution position of each synthetic peptide without any detectable immunoreactivities at high molecular weight positions. These results indicate the presence of Met-Enk-Arg6-Gly7-Leu8 together with Met-Enk, Leu-Enk and Met-Enk-Arg6-Phe7 in human gastric antrum and further suggest that these opioid peptides are derived from the same preproenkephalin as that in the adrenal medulla and that the processing of preproenkephalin is almost complete in the gut.

Three cases of primary hyperparathyroidism associated with nonmedullary thyroid carcinoma.

Nonmedullary thyroid carcinoma was diagnosed in three of nine cases of primary hyperparathyroidism. In all three cases, diagnosis of primary hyperparathyroidism was made before that of thyroid carcinoma. In the first case, follicular carcinoma was incidentally detected during parathyroidectomy. In the second case, thyroid tumor was discovered during imaging studies for hyperparathyroidism. Papillary carcinoma and ectopic parathyroid was diagnosed postoperatively. In the third case, a thyroid lesion had been mistaken for a putative parathyroid lesion, but the diagnosis of papillary carcinoma was obtained. These cases suggest that preoperative examination for hyperparathyroidism should be carefully evaluated, considering possible concomitant thyroid lesions.

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.

Rhytidectomy in orientals.

The technique of rhytidectomy used by the authors for the oriental face is explained. The main points of this operative procedure are: 1) the incision line is designed before the operation, 2) subcutaneous undermining is easy, 3) the key sutures are done between the auricular cartilage and the cheek skin to prevent wide scars and, 4) the special incision lines are designed to prevent the downward deformation of the earlobe. This paper also discusses the anatomical differences between Japanese and Caucasian rhytidectomy.

[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.

[Cytological and immunocytochemical study of prostatic carcinoma cell in voided urine. Usefulness of prostate-specific antigen in cytology specimen].

The presence of prostatic carcinoma cells in voided urine was seldom noticed and these cells might be hardly distinguished from transitional cell carcinoma cells of the urinary tracts on the Papanicolaou-stained slides. We investigated the cytological findings and the localizations of prostate-specific antigen (PSA) in 7 cases of prostatic carcinomas (4 adenocarcinomas, 2 squamous cell carcinomas, and 1 undifferentiated carcinoma) which showed tumor cells in voided urine. The cytological findings of each histological types were as follows: adenocarcinoma cells were observed in small clusters of round or columnar cells with eccentric nuclei and prominent nucleoli with occasion, squamous cell carcinoma cells were characteristically well-differentiated keratinizing ones varying in shape and in staining reaction, and undifferentiated carcinoma cells were polyhedral and large ones with dense cytoplasm. After destaining of the Papanicolaou-stained slides, the positivity for PSA was demonstrated by immunocytochemical stain with PSA antiserum in cytoplasms of the tumor cells of 6 cases except for 1 undifferentiated carcinoma. We recognized that immunocytochemical detection of PSA was a useful mean in cytological detection for prostatic carcinomas in voided urine.

[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.

[Clinicopathological study on microscopic urinalysis as screening for malignant cells].

Study on detection of malignant cells in urinary sediments using supravital staining was described. We examined 96,554 specimens of urinary sediments for 2 years from January 1985 to December 1986. The results of microscopic urinalysis were compared with the cytological and histological diagnoses. Atypical cells were found in 138 patients, and 47 (34.1%) cancers were diagnosed histologically among them. These included 33 bladder cancer, 1 ureter cancer, 1 renal pelvic cancer, 2 prostate cancers, 1 rectal cancer, and 9 uterine cancers. Seven patients of them had not been under suspicion of malignancy yet before atypical cells were detected. Therefore microscopic urinalysis caused the triggers of cancer diagnoses. For bladder cancers, the positive rates in microscopic urinalysis were 43.4%, and those in urinary cytology were 52.4%. The positivity revealed higher in high-grade cancers than in low-grade. As compared with the results between microscopic urinalysis and urinary cytology in identical patients, the rate of correspondence between them was 89.5%. In 61.2% of positive and suspicious urinary cytology, atypical cells were not found. Atypical cells were seen in negative urinary cytology of 26 cases, and 5 cases of cancers were diagnosed histologically. These suggested that microscopic urinalysis as a screening for malignant cells was useful to detect urinary tract malignancy combining with urinary cytology.

[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.

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.