Persistent sodium currents in mesencephalic v neurons participate in burst generation and control of membrane excitability

The functional and biophysical properties of a persistent sodium current (I(NaP)) previously proposed to participate in the generation of subthreshold oscillations and burst discharge in mesencephalic trigeminal sensory neurons (Mes V) were investigated in brain stem slices (rats, p7-p12) using whole cell patch-clamp methods. I(NaP) activated around -76 mV and peaked at -48 mV, with V1/2 of -58.7 mV. Ramp voltage-clamp protocols showed that I(NaP) undergoes time- as well as voltage-dependent inactivation and recovery from inactivation in the range of several seconds (tau(onset) = 2.04 s, tau(recov) = 2.21 s). Riluzole (< or =5 microM) substantially reduced I(NaP), membrane resonance, postinhibitory rebound (PIR), and subthreshold oscillations, and completely blocked bursting, but produced modest effects on the fast transient Na+ current (I(NaT)). Before complete cessation, burst cycle duration was increased substantially, while modest and inconsistent changes in burst duration were observed. The properties of the I(NaT) were obtained and revealed that the amplitude and voltage dependence of the resulting "window current" were not consistent with those of the observed I(NaP) recorded in the same neurons. This suggests an additional mechanism for the origin of I(NaP). A neuronal model was constructed using Hodgkin-Huxley parameters obtained experimentally for Na+ and K+ currents that simulated the experimentally observed membrane resonance, subthreshold oscillations, bursting, and PIR. Alterations in the model g(NaP) parameters indicate that I(NaP) is critical for control of subthreshold and suprathreshold Mes V neuron membrane excitability and burst generation.     

Association of bone mineral density with vitamin D and estrogen receptor gene polymorphisms during GnRH agonist treatment

AIMS: This study examined whether or not a decrease in bone mineral density (BMD) induced by the use of gonadotropin-releasing hormone agonist (GnRHa) during sexual maturation is affected by vitamin D receptor and/or estrogen receptor gene polymorphisms, like the phenomenon observed during the postmenopausal period. METHODS: In 43 patients who received GnRHa therapy for 6 months to treat uterine myoma or endometriosis at our department and who were confirmed to have pituitary down-regulation, we measured bone density before and after GnRHa treatment using DXA and analyzed the bone metabolism turnover using bone metabolic markers. Polymorphisms were analyzed by RFLP using FokI and TaqI for the vitamin D receptor gene and PvuII and XbaI for the estrogen receptor gene. The then determined gene polymorphism was analyzed in relation to the percentage decreases in BMD following GnRHa treatment. RESULTS: The patients were divided by f, t into two groups: (f, t) < 2 (Group V-I) and (f, t) > or = 2 (Group V-II). They were also divided by P, x into two groups (P, x) < 3 (Group E-I) and (P, x) > or = 3 (Group E-II). The BMD change was significantly higher in Group V-II than in Group V-I. Group E-II tended to have a higher BMD change than Group E-I, although this difference was not statistically significant. CONCLUSION: Patients who often have f and t polymorphism are more likely to show BMD reduction following GnRHa therapy, like the phenomenon seen during the postmenopausal period, than patients with other gene polymorphisms. Measures to avoid BMD reduction are required when using GnRHa in such patients.     

Successful treatment for advanced hepatocellular carcinoma by hepatic arterial infusion of CDDP powder as second-line chemotherapy

A 72-year-old man with type C liver cirrhosis had suffered from hepatocellular carcinoma (HCC) since April, 2001. HCC spread diffusely all over the right lobe of his liver, and the serum alpha-fetoprotein (AFP) value increased up to 42,696 ng/ml in June of 2004. He was implanted with a port-catheter system, and hepatic arterial infusion chemotherapy (HAIC) using low-dose of CDDP and 5-FU was started. However, it was not effective and after 4 months, the serum AFP level increased up to 755,030 ng/ml, ascites appeared, and gastro-esophageal varices also spread. No definite metastasis was detected, then we started to second-line chemotherapy. He was then given HAIC using CDDP powder for intraarterial use (CDDP 50 mg/m(2)/20 min, monthly). After 3 courses, the serum AFP level decreased to 9 10 ng/ml, and abdominal CT revealed that the main tumor had regressed and ascites had disappeared. After one more course, the serum AFP value decreased to 8 ng/ml and complete response was achieved on abdominal CT imaging. There was no major complication related to the chemotherapy. HAIC for advanced HCC using LFP has been reported to achieve favorable results, but no other regimens have been proved to the standard for HCC. HAIC using CDDP powder for advanced HCC may be beneficial as the second-line chemotherapy.     

E148Q/M694I mutation in 3 Japanese patients with familial Mediterranean fever

We describe 3 unrelated Japanese patients with familial Mediterranean fever (FMF) due to a compound heterozygous E148Q/M694I mutation in the MEFV gene. The first patient is a 38-year-old man who also has chronic myelogenous leukemia (CML). Because genomic DNA analysis of the patient's nail revealed the E148Q/M694I mutation, we concluded that the individual mutations were obtained congenitally. Interferon alpha therapy was effective against not only the CML but also the FMF. The second patient is a 42-year-old man with consanguineous parents and a 14-year history of recurrent lower abdominal and back pain associated with fever. He successfully responded to colchicine treatment. The third patient is a 23-year-old woman who has a family history of FMF and since the age of 11 years has had recurrent chest and abdominal pain with fever. The onset of FMF was at an early age in this case, in contrast with the late onset of the disease in the first 2 cases. This patient's mother also has a heterozygous M694I mutation and experienced the same symptoms until 30 years of age. Our data suggest that it should be recognized that there are more FMF patients in Japan than previously expected and that the frequency of the E148Q/M694I mutation may be significant in Japanese FMF patients.     

Helicobacter pylori stool antigen test

In Japan, Helicobacter pylori stool antigen(HpSA) test was opened for diagnosis of H. pylori infection before and after eradication therapy form Nov. 1, 2003. We can use two kinds of HpSA kits, Premier Platinum HpSA(Meridian Bioscience, USA) and Testmate pylori antigen EIA(Wakamoto Pharmaceutical Co., Ltd., Japan). We evaluated the diagnostic accuracy of the HpSA test(Premier Platinum HpSA) compared with diagnosis based on endoscopic biopsy-based methods. In 136 patients, the sensitivity and specificity of the HpSA test before therapy were 98.3% and 95.0%, respectively. In 54 patients, the sensitivity and specificity of the HpSA test after eradication therapy were 90% and 97.7%, respectively. Therefore, it is concluded that the HpSA test is a useful method for the diagnosis of H. pylori infection before and after eradication therapy.     

Subretinal administration of tissue-type plasminogen activator to speed the drainage of subretinal hemorrhage

• Background: Bleeding into the subretinal space in the vicinity of the macula is associated with age-related macular degeneration or retinal arterial macroaneurysm. The prognosis for restoration of vision is poor in the presence of blood clots. • Methods: Using a simple device composed of three disposable syringes we injected tissue-type plasminogen activator (tPA) into the subretinal space during conventional vitrectomy in six patients to assist the draining of subretinal clots. • Results: Four of six patients recovered their visual acuity postoperatively, while visual acuity in the other patients was stabilized. • Conclusion: Early drainage of subretinal hemorrhage assisted by the introduction of tPA into the subretinal space led to uncomplicated surgery and favorable postoperative results. Received: 14 January 1997 Revised version received: 6 May 1997 Accepted: 26 May 1997     

Water absorption of poly(methyl methacrylate) containing 4-methacryloxyethyl trimellitic anhydride

The amount of water absorption of poly(methyl methacrylate) (PMMA) containing 0, 1, 3 and 5 wt% of an adhesive monomer, 4-methacryloxyethyl trimellitic anhydride (4-META), was measured at 7 degrees C, 37 degrees C and 60 degrees C. After the water uptake reached equilibrium in specimens, they were desorbed to obtain a constant value and the absorption process was repeated. Mass changes in the second desorption were recorded for the storage temperatures of 37 degrees C and 60 degrees C. Multiple regression analyses were conducted on three independent variables, 4-META concentration, storage temperature and absorption-desorption cycle. A statistically significant relationship was found between the maximum water uptake and 4-META concentration, while there was no relationship between the maximum water uptake and diffusion coefficient obtained using the Fick's law. The negative relationship in the latter did not support the free space theory. The significant and positive relationship between the maximum water uptake and 4-META concentration demonstrates that water molecules diffuse through the formation of a hydrogen bond at polar sites. The maximum water uptake was not influenced by temperature, while the diffusion coefficient increased with the rise in temperature. The activation energy was 41-47 and 50-53 kJ/mol in the first and second absorption tests, respectively.     

The effect of medial collateral ligament insufficiency on the reconstructed anterior cruciate ligament: a study in the rabbit

The treatment for severe combined anterior cruciate ligament (ACL) and medial collateral ligament (MCL) ruptures is disputed. Using a rabbit model, we examined the effect of insufficiency of medial structures on the reconstructed ACL in combined ACL and MCL injury. 40 rabbits were divided into 2 groups. In both groups, ACL was subjected to in situ freeze-thaw treatment. In group F, only freeze-thaw treatment of ACL was given. In group FM, partial resection of MCL was also done. We killed 5 rabbits on each of 4 occasions: immediately after the operation (time 0), at 6, 12 and 24 weeks postoperatively. At each time, we measured valgus instability and mechanical properties of the ACL. Valgus instability in group FM persisted from time 0 to 24 weeks, and was significantly greater than that in group F. The tensile strength and tangent modulus of the ACL in group FM were lower than those in group F. We found that continuous valgus instability reduces the mechanical properties of the in situ frozen ACL.     

Results of a phase I clinical study using autologous tumour lysate-pulsed monocyte-derived mature dendritic cell vaccinations for stage IV malignant melanoma patients combined with low dose interleukin-2

We conducted a pilot study to assess the feasibility and efficacy of immunotherapy for stage IV malignant melanoma patients resistant to conventional therapies involving vaccination with mature dendritic cells (mDCs) combined with administration of low dose interleukin-2. Autologous monocytes were harvested from a single apheresis and cultured for 7 days with granulocyte-macrophage colony-stimulating factor and interleukin-4, yielding immature dendritic cells (iDCs), which were then cryopreserved until use. For 4 days prior to vaccination, iDCs were exposed to autologous tumour lysate combined with tumour necrosis factor-alpha to induce terminal differentiation into mDCs. Patients were then vaccinated weekly with 107 mDCs for 10 weeks and given 350-700 kIU of interleukin-2 three times per week. Of the 10 patients in the study, one showed stable disease, seven showed progressive disease, and two showed mixed responses, including partial tumour regression, and were therefore given 20 additional injections. Only minimal adverse events were noted, including localized skin reactions and mild fever (NIH-CTC grade 0-1). Median survival from the first vaccination was 240 days (range 31-735 days). In vitro, melanoma patient-derived dendritic cells (DCs) showed reduced cell surface expression of CD1a antigen on iDCs and reduced CD86 and HLA-DR expression on mDCs. In addition, antigen uptake, chemotaxis and antigen presentation were all attenuated in DCs from the patients. In summary, although improvement of clinical efficacy will require further research, autologous tumour lysate-pulsed monocyte-derived mDCs could be safely harvested, cryopreserved and administrated to patients without obvious complications.     

Bilateral elastofibroma of the thighs with concomitant subscapular lesions

We report a case of elastofibroma that occurred in both thighs and subscapular regions. The MR signal intensity of the lesions was typical for elastofibroma. Although bilateral involvement of the thighs is exceptional, the presence of concomitant characteristic bilateral subscapular lesions was helpful in making the diagnosis of elastofibroma.