Effect of pilocarpine on substance P and calcitonin gene‐related peptide releases correlate with salivary secretion in human saliva and plasma

What is known and Objective:  Pilocarpine, a muscarinic receptor agonist, has been used for the treatment of dry mouth. Salivary glands are supplied with nerve fibres that contain neuropeptides, such as substance P, calcitonin gene‐related peptide (CGRP) and vasoactive intestinal polypeptide (VIP), which are important modulators of salivation. It is known that measurement of salivary and plasma levels of neuropeptides is useful for assessing the dose–pharmacological effect relationship of drugs. The relationship between the action of pilocarpine and neuropeptides in humans has not been studied. Moreover, studies evaluate the usefulness of drug salivary levels in the pharmacological evaluation of drugs are scarce. The aim of this study was to examine the effects of pilocarpine on the levels of substance P‐, CGRP‐ and VIP‐like immunoreactive substances (IS) in saliva and plasma taken in healthy humans. Methods:  Five healthy male subjects participated in this study. Pilocarpine tablet (10 mg) or placebo tablet was orally administered with 100 mL of water. Each subject was administered placebo and drug with an interval of 4 weeks in between. Saliva was sampled before and at 20, 40, 60, 90, 120, 180 and 240 min after administration of the test substances. Venous blood samples (10 mL) were also taken from a forearm vein at each time interval. The samples were then enzyme immunoassayed using a highly sensitive system for substance P‐, CGRP‐ and VIP‐IS. The amount of saliva was measured by the Saxon test. Results:  A single oral administration of pilocarpine increased the release of salivary substance P‐IS (the area under the concentration‐time curve: AUC_{0→240 min}) compared with the placebo. Pilocarpine also significantly increased the release of salivary CGRP‐IS (AUC_{0→240 min}). Pilocarpine significantly increased the release of plasma CGRP‐IS. The salivary volume correlated with the salivary level of substance P and CGRP‐IS (r = 0·84, P < 0·05 and r = 0·59, P < 0·05, respectively). AUC_{0→240 min} for substance P‐IS in saliva correlated with that for plasma (r = 0·78, P < 0·05). What is new and Conclusion:  Pilocarpine increases the release of salivary substance P and CGRP‐IS. This suggests that one mechanism by which pilocarpine improves dry mouth is by local stimulation of neuropeptidergic nerves. Moreover, saliva levels of substance P showed good correlation with the plasma levels. The substance P levels in saliva and plasma may be good indicators of the effects of drugs used in dry mouth/xerostomic patients.     

Detection and characterization of individual endocytosis of AMPA‐type glutamate receptor around postsynaptic membrane

Synaptic plasticity such as long‐term depression (LTD) has been regarded as a cellular mechanism of learning and memory. LTD is expressed by the decrease in number of postsynaptic AMPA‐type receptor (AMPAR) at glutamatergic synapses. Although endocytosis is known to play an essential role in the decrease in AMPAR on postsynaptic membrane, the difficulty to detect individual endocytic events hampered clarification of AMPAR dynamics around synapses. Previously, we developed a method to induce formation of postsynaptic‐like membrane (PSLM) on the glass surface and observed pHluorin‐tagged AMPAR around PSLM with total internal reflection fluorescence microscopy. By this method, individual exocytosis of AMPAR‐pHluorin was recorded in both PSLM and non‐PSLM. In other studies, endocytic vesicles containing pHluorin‐tagged receptors were visualized by changing extracellular pH. Here, we have combined PSLM formation method and rapid pH change method, and detected individual endocytic events of AMPAR around PSLM with high spatial and temporal resolutions. Endocytic events of AMPAR were characterized by comparison with those of transferrin receptor. Constitutive endocytosis of AMPAR was not dependent on clathrin and dynamin in contrast to that of transferrin receptor. However, AMPAR endocytosis triggered by LTD‐inducing stimulation was clathrin‐ and dynamin‐dependent.     

Utility of the new Japanese severity score and indications for special therapies in acute pancreatitis

Background  The Japanese severity score (JSS) for acute pancreatitis was revised in 2008. As special therapies for severe acute pancreatitis (SAP), continuous regional arterial infusion of protease inhibitor and antibiotics (CRAI) and enteral nutrition (EN) are now utilized in Japan. We investigated the usefulness of the new JSS and the indications for CRAI and EN based on the new JSS. Methods  We assessed the new JSS in 138 patients with SAP according to the previous Japanese criteria. Usefulness of the new JSS for the prediction of mortality rates was compared with conventional scoring systems by receiver-operator characteristic curve analysis. We analyzed the relationship between the new JSS and prognosis in patients with and without CRAI and EN, respectively. Results  Forty-five patients (33%) were assessed as having mild acute pancreatitis, and 93 patients (67%) were assessed as having SAP. Their mortality rates were 7 and 40%, respectively. The area under the curve for the prediction of mortality rates with the new JSS was 0.822 and was the highest among conventional scoring systems. In patients with new JSS ≥ 6, the mortality rate was lower in patients with CRAI than in patients without CRAI (P = 0.129). In patients with new JSS ≥ 4, the mortality rate was lower in patients with EN than in patients without EN (P = 0.016). Conclusions  The new JSS is useful and easier to use for the prediction of prognosis compared to the conventional scoring systems. EN was effective in reducing the mortality rate in patients with a new JSS ≥ 4.     

Cross‐sectional analysis of germline BRCA1 and BRCA2 mutations in Japanese patients suspected to have hereditary breast/ovarian cancer

The prevalence of BRCA1/2 germline mutations in Japanese patients suspected to have hereditary breast/ovarian cancer was examined by a multi‐institutional study, aiming at the clinical application of total sequencing analysis and validation of assay sensitivity in Japanese people using a cross‐sectional approach based on genetic factors estimated from personal and family histories. One hundred and thirty‐five subjects were referred to the genetic counseling clinics and enrolled in the study. Full sequencing analysis of the BRCA1/2 gene showed 28 types of deleterious mutations in 36 subjects (26.7%), including 13 types of BRCA1 mutations in 17 subjects (12.6%) and 15 types of BRCA2 mutations in 19 subjects (14.1%). Subjects were classified into five groups and 22 subgroups according to their personal and family history of breast and/or ovarian cancer, and the prevalence of deleterious mutations was compared with previously reported data in non‐Ashkenazi individuals. Statistical analysis using the Mantel‐Haenszel test for groups I through IV revealed that the prevalence of Japanese subjects was significantly higher than that of non‐Ashkenazi individuals (P = 0.005, odds ratio 1.87, 95% confidence interval 1.22–2.88). Family history of the probands suffering from breast cancer indicated risk factors for the presence of deleterious mutations of BRCA1/2 as follows: (1) families with breast cancer before age 40 within second degree relatives (P = 0.0265, odds ratio 2.833, 95% confidence interval 1.165–7.136) and (2) families with bilateral breast cancer and/or ovarian cancer within second degree relatives (P = 0.0151, odds ratio 2.88, 95% confidence interval 1.25–6.64). (Cancer Sci 2008; 99: 1967–1976)     

Clear-cell adenocarcinoma of the uterine cervix in a 17-year-old adolescent

This report describes the case of the youngest Japanese person to be diagnosed with endocervical clearcell adenocarcinoma. In September 2005, a 17-year-old female adolescent visited a physician because of vaginal bleeding. A cervical tumor was discovered, and the patient was referred to our outpatient department. Vaginal examination showed a bleeding tumor approximately 1.5 cm in size protruding from the cervical os. The cytological finding of the uterine cervix was positive for malignancy, and the histological diagnosis by punch biopsy was clear-cell adenocarcinoma of the uterine cervix. A radical abdominal hysterectomy and pelvic lymphadenectomy were performed on October 30. Macroscopic findings showed a tumor, 1.5 cm in diameter, growing from the right side of the uterine cervix. The pathological diagnosis was clear-cell adenocarcinoma of the cervix (PT1b1NR0M0). The patient was discharged from our hospital without any adjuvant therapy. No signs of recurrence have been detected in the 2-year follow up.     

Characterization and clinical evaluation of tumor-associated antigen CA54/61 identified by monoclonal antibodies MA54 and MA61 in epithelial ovarian cancer

Monoclonal antibodies (moABs) MA54 and MA61, directed toward the O-linked mucin-type glycoprotein, have been established and showed highly specific reactivity with human ovarian cancer. Fetal intestinal and colonic mucosal cells expressed this antigen and meconium staining was also frequently positive. To investigate the characteristic of an epitopic carbohydrate recognized by these moABs, the reactivity of each moAB with meconium extract was monitored by solid-phase enzyme-linked immunosorbent assay with mono-, di-, and oligosaccharides. MA54 and MA61 react with meconium extract and the reactivities of these moABs are neuraminidase sensitive. Ovine submaxillary mucin had a strong inhibitory activity toward the reaction between meconium extract and MA54 as well as MA61, suggesting that these moABs recognize NeuAc 2-6GalNAc epitope in meconium. The second aim of this study is to investigate the possible application of moABs to diagnose ovarian cancer and to compare these levels with those of the CA125 antigen. While serum CA54/61 antigen levels were elevated in 44.4% of ovarian cancer cases and serum CA125 antigen levels were elevated in 86.7% of the same population, the use of both assays indicated a sensitivity of detection of 97.8% (44 of 45 patients) in the population studied.     

The effect of crossed olivo-cochlear bundle stimulation on acoustic trauma.

To investigate whether the crossed olivo-cochlear bundle (COCB) functions in a protective manner, albino guinea pigs were exposed to sounds of varying intensity (110-130 dB SPL, 3-30 min) with or without electric stimulation of COCB, and the threshold shifts of the compound action potential (CAP) were examined. A statistically significant protective effect was observed in animals exposed to stimuli of intermediate intensity which induce threshold shifts of 50 to 55 dB on average. No protective effect was observed in the groups exposed to greater or milder stimuli. These results are discussed in the light of the available literature.