Novel anti-idiotype antibody therapy for lipooligosaccharide-induced experimental autoimmune neuritis: use relevant to Guillain-Barré syndrome.

Campylobacteriosis is a frequent antecedent event in Guillain-Barré syndrome (GBS), inducing high-titer serum antibodies for ganglioside antigens in the peripheral nervous system (PNS). Molecular mimicry between the lipooligosaccharide (LOS) component of Campylobacter jejuni and human peripheral nerve gangliosides is believed to play an important role in the pathogenesis of GBS. Conventional treatment strategies for patients with GBS include plasmapheresis, intravenous immunoglobulin (IVIG), and immunosuppression, which are invasive or relatively ineffective. In this study, we used our animal model of GBS, in which Lewis rats were immunized with GD3-like LOS isolated from C.jejuni. The animals developed anti-GD3 ganglioside antibodies and manifested neuromuscular dysfunction. To develop novel therapeutic strategies, we treated the animals by intraperitoneal administration of an anti-GD3 antiidiotype monoclonal antibody (BEC2) that specifically interacts with the pathogenic antibody. The treated animals had a remarkable reduction of anti-GD3 antibody titers and improvement of motor nerve functions. The results suggest that ganglioside mimics, such as antiidiotype antibodies, may be powerful reagents for therapeutic intervention in GBS by neutralizing specific pathogenic antiganglioside antibodies.

Safety and Effectiveness of Conversion From Cyclosporine to Once-Daily Prolonged-Release Tacrolimus in Stable Kidney Transplant Patients: A Multicenter Observational Study in Japan.

This study investigated the safety and effectiveness of conversion from cyclosporine- to prolonged-release tacrolimus (PR-T)-based immunosuppression in kidney transplant recipients (KTRs) in Japanese routine clinical practice. MATERIALS AND METHODS: This was a prospective observational study of stable KTRs who were converted from cyclosporine to PR-T according to local clinical practice. Clinical data were collected up to 12 months postconversion. Study outcomes included conversion dosing ratios, PR-T dose and trough levels, change in estimated glomerular filtration rate between conversion and month 12, graft/patient survival, and rejection rate (Kaplan-Meier). Outcomes of ongoing preconversion hypertrichosis, gingival hypertrophy, and cyclosporine-related renal toxicity were detailed. Data for adverse drug reactions were collected. RESULTS: Overall, 266 patients (mean ± SD age 51.9 ± 13.5 years) were included. The mean ± SD conversion ratio (PR-T:cyclosporine, mg:mg) was 0.029 ± 0.017. After an initial decrease between conversion and month 3, mean ± SD PR-T daily dose remained stable up to month 12 (2.4 ± 1.5 mg at months 3 and 12), as did tacrolimus trough blood levels (3.5 ± 1.8 vs 3.6 ± 1.7 ng/mL, respectively). Estimated glomerular filtration rate was stable over 12 months (mean ± SD change from conversion to month 12 was 0.3 ± 7.8 mL/min/1.73m2). Month 12 Kaplan-Meier patient and graft survival rates were 99.6% and 95.5%, respectively. Eight patients reported 9 rejection episodes. PR-T demonstrated potential to improve cyclosporine-related renal toxicity, hypertrichosis, and gingival hypertrophy. Postconversion, 46 adverse drug reactions were reported in 39 patients (14.7%); there was 1 death. CONCLUSIONS: Conversion from cyclosporine to PR-T in Japanese stable KTRs was effective and tolerable over 12 months, with low rates of rejection reported.

Effect of dithiothreitol on angiotensin II receptor type II in rat ovarian cultured granulosa cells.

Dithiothreitol markedly increased the ligand binding affinity of angiotensin II (AII) receptor type II (AT2) without affecting its antagonist selectivity in cultured ovarian granulosa cells, demonstrating that this AT2 is of the dithiothreitol-sensitive type. Dithiothreitol is useful for specifically detecting low levels of the AT2 in the ovary, where it plays roles that are probably related to atresia.

Increase in DNA polymerase alpha activity associated with DNA synthesis due to FSH or oestrogen in ovaries of immature rats.

DNA polymerase activities and DNA content of ovaries from immature intact rats (4-29 days after birth), hypophysectomized rats and hormone-treated hypophysectomized rats were measured. During normal ovarian growth DNA polymerase alpha activity and DNA content of ovaries increased. The polymerase activity decreased gradually after hypophysectomy without any alteration in the DNA content. Administration of ovine FSH (2 micrograms/day) or oestradiol-17 beta (1 mg/day) to hypophysectomized rats enhanced ovarian DNA content and DNA polymerase alpha activity, whereas DNA polymerase beta activity did not change significantly. These results suggest that DNA polymerase alpha participates in DNA synthesis in these ovaries. The specific activity of DNA polymerase alpha (the activity per microgram DNA) in the ovaries increased between 4 and 14 days after birth, and then remained almost constant; the specific activity declined gradually after hypophysectomy. Administration of FSH or oestradiol-17 beta but not of ovine LH, progesterone or testosterone to hypophysectomized rats restored the specific activity. Mixing experiments with different kinds of ovarian extracts suggested that no activators of DNA polymerase alpha were present in the extracts. These results suggest that FSH or oestrogen causes the induction of DNA polymerase alpha accompanied by DNA synthesis during cell proliferation in ovaries of immature rats.

Endothelin-1 in luteal tissue.

The aim of this study was to immunologically and biologically detect endothelin-1 (ET-1) in rat corpora lutea (CL). Recently, we established a highly sensitive and specific sandwich-enzyme immunoassay (EIA) for ET-1. Using this assay, the presence of ET-1 was investigated in superovulated ovaries, induced with pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG), and ovaries from pseudopregnant rats, induced by cervical stimulation. A high concentration of immunoreactive endothelin-1 (ir-ET-1) was found in the CL. On reverse phase-high performance liquid chromatography (RP-HPLC) coupled with EIA, ir-ET-1 was exclusively eluted at the same position as synthetic ET-1, indicating that ir-ET-1 is identical to ET-1. The level of ir-ET-1 was significantly (P less than 0.001) higher in the CL 7 days after hCG injection than it was 4 days after hCG injection. On day 7 of pseudopregnancy (PSP), the ir-ET-1 level was also significantly (P less than 0.001) higher than on day 4 of PSP. These results demonstrated that ET-1 is present in a high concentration in the CL, suggesting a new intraovarian peptide which may have a physiological function in the ovary and which may vary in quantity according to the age of the CL.

Stimulation of glycosphingolipid biosynthesis by L-threo-1-phenyl-2-decanoylamino-1-propanol and its homologs in B16 melanoma cells.

Previous studies have demonstrated that the ceramide analog D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-threo-PDMP) inhibits glucosylceramide (GlcCer) synthase and thus leads to extensive depletion of glycosphingolipids (GSLs) biosynthesized from GlcCer [reviewed by Radin, N.S., Shayman, J.A., and Inokuchi, J. (1993) Adv. Lipid Res. 26, 183-213). In the present study, stereospecificity of PDMP activity was demonstrated with an enantiomeric pair, D-threo-PDMP and L-threo-PDMP. Treatment of B16 melanoma cells with the D-threo or L-threo isomer produced contrasting changes of GSL biosynthesis, as monitored by metabolic labeling with [3H]Gal. D-PDMP markedly inhibited incorporation of radioactivity into GlcCer, LacCer, and GM3 as expected, whereas the L-threo isomer significantly increased it. Homologs of L-PDMP having different N-acyl chains were synthesized and also tested for their effects. Among them, the compounds having C8-C14 acyl chains increased incorporation of the radioactivity into GSLs to different degrees, demonstrating that the stimulatory effect of the L-threo homologs depends on acyl chain length. In order to elucidate the biochemical mechanisms of these PDMP effects, the activities of GlcCer synthase, LacCer synthase, and GM3 synthase in B16 cell lysates were measured in the presence of PDMP. D-Threo-PDMP but not the L-threo isomer inhibited both LacCer and GM3 synthases as well as GlcCer synthase, suggesting that the ceramide-like structure of the D-PDMP molecule interacted stereospecifically with these GSL-synthesizing enzymes. On the other hand, L-PDMP had no effect in the in vitro assays.(ABSTRACT TRUNCATED AT 250 WORDS)

A synthetic ceramide analog (L-PDMP) up-regulates neuronal function.

To address the role of brain gangliosides in synaptic activity, the ceramide analogs, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) and its enantiomer, L-PDMP, were used to inhibit and stimulate ganglioside biosynthesis in cultured cortical neurons. Prolonged treatment with both PDMP isomers exhibited opposite effects on functional synapse formation measured by spontaneous synchronized oscillatory activity of intracellular Ca2+ between the neurons: suppression by D-PDMP and facilitation by L-PDMP. Up-regulation of synaptic activity by L-PDMP could be correlated with the slow but robust activation of p42 mitogen-activated protein kinase. Treatment with L-PDMP after transient forebrain ischemia in rats ameliorated the deficit of a well-learned spatial memory by an 8-arm maze task, suggesting a new potential therapeutic approach for neurodegenerative disorders.

Follicle-stimulating hormone induces deoxyribonucleic acid polymerase alpha before deoxyribonucleic acid synthesis in immature rat ovarian follicles perfused in vitro.

To examine the effects of follicle-stimulating hormone (FSH) on cell proliferation and cell division in ovaries, deoxyribonucleic acid (DNA) polymerase activities and DNA concentrations were determined in ovarian follicles of immature, hypophysectomized, estradiol-treated rats perfused in vitro. Ovine FSH (2-20 micrograms/mL) significantly enhanced the DNA polymerase alpha activity in the follicles perfused for 180-300 minutes, whereas the DNA polymerase beta activity and DNA concentration showed no significant changes. It is concluded that FSH causes the induction of DNA polymerase alpha before DNA synthesis during follicle growth in immature rat ovaries.

Rat corpus luteum expresses both PACAP and PACAP type IA receptor mRNAs.

Both pituitary adenylate cyclase-activating polypeptide (PACAP) and PACAP type I receptor gene expressions were detected in the corpus luteum of pregnant mare's serum gonadotropin (PMSG)-human chorionic gonadotropin (hCG)-treated immature rats using reverse transcription-polymerase chain reaction (RT-PCR). RT-PCR products of the poly(A)+ RNA extracted from rat corpora lutea yielded dominant DNA bands that corresponded to segments of PACAP mRNA (453 bp) and PACAP type IA receptor mRNA (290 bp) spanned by the PCR primers. The identities of the PACAP cDNA and the PACAP receptor cDNA fragments were confirmed by Southern blot hybridization analyses. Our results showed that PACAP mRNA and PACAP type IA receptor mRNA are synthesized within luteinized cells of rat ovary, and suggest that PACAP is closely linked to the reproductive process.

[Secretary sialidase activity and GM3 ganglioside].

The sialidase activities with GM3 ganglioside and sialyllactitol were demonstrated in the conditioned medium of human fibroblasts. pH versus activity profiles of conditioned medium with GM3 as substrate suggested the presence of two sialidases with optimal activities at pH 4.5 and pH 6.5. The GM3 sialidase activity at pH 6.5 was suppressed in the medium of contact-inhibited cells. This sialidase may function in the metabolism of cell surface GM3 since there was a selective loss of labeled sialic acid from GM3 at different times of incubation after pulse-labeling with a radioactive sialic acid precursor ([3H]N-acetyl-mannosamine) and a radioactive ceramide precursor ([14C]serine). In addition, a sialidase inhibitor, 2-deoxy-2, 3-dehydro-N-acetyl-neuraminic acid (NeuAc-2-en) resulted in a reversible growth inhibitory effect and the suppression of the sialidase activity in the medium. We have speculated that GM3 hydrolysis on the cell surface by the sialidase may be coordinated with the cell cycle and may be at its maximum during early in the G1 phase.

Case report: thyrotropin-releasing hormone-induced myoclonus and tremor in a patient with Hashimoto's encephalopathy.

The authors investigated the possibility of a thyrotropin-releasing hormone-related mechanism in a 43-year-old Japanese woman with Hashimoto's encephalopathy who experienced three relapses closely associated with the menstrual cycle. Her symptoms began at ovulation, worsened during the luteal phase, and improved during the menstruation phase. No abnormalities were found by brain magnetic resonance imaging and cerebral angiography. Intravenous administration of thyrotropin-releasing hormone induced symptoms of myoclonus and tremor similar to those observed during an exacerbation. The intensity and duration of involuntary movements induced by thyrotropin-releasing hormone were dose-dependent. The patient's symptoms were controlled effectively by thyroxine replacement therapy. On the basis of these findings, thyrotropin-releasing hormone may have an important role in Hashimoto's encephalopathy.

Tokishakuyakusan stimulates progesterone and estradiol-17 beta production by rat granulosa cells and progesterone, testosterone and estradiol-17 beta by the residual portion of the follicle in vitro.

To examine the possible effects of Tokishakuyakusan (TS) on steroidogenesis by preovulatory follicles at the cell level, the expressed granulosa cells and remaining portion of follicles from pregnant mare's serum gonadotropin (PMS)-treated immature rats were incubated in vitro with increasing concentrations of TS for 3 h. TS significantly stimulated progesterone and estradiol-17 b production, with a predominant stimulation of progesterone, by the expressed granulosa cells, while testosterone production was not stimulated. In the remaining portion of the follicle, TS also significantly stimulated progesterone, testosterone and estradiol-17 b production. Similar to the effect produced by granulosa cells, the stimulatory effect of TS was stronger on progesterone than on testosterone and estradiol-17 b production. These results suggest that TS has a potent, direct stimulatory effect on steroidogenesis, especially progesterone production, by constituent tissue compartments of rat preovulatory follicles in vitro.

The direct luteotropic effect of tokishakuyakusan in rats.

The effect of Tokishakuyakusan (TS) on the corpus luteum function in pseudopregnant rats was examined in vivo. On day 4 of pseudopregnancy (PSP), induced by cervical stimulation, TS (20 micrograms) stimulated the progesterone secretion rate (PSR) in the ovarian venous plasma. There was also a significant increase in the rate of progesterone to 20 alpha-OH-progesterone. However, on day 8 of PSP, there was no apparent change in PSR in the ovarian venous plasma after the administration of TS. These data suggest that the sensitivity to TS of the corpus luteum varies according to its age.

Effect of tokishakuyakusan on corpus luteum endothelin.

The presence of endothelin-1 (ET) and effect of Tokishakuyakusan (TS) on ET in rat corpora lutea (CL) was investigated in superovulated ovaries, induced with pregnant mare's serum gonadotropin and human chorionic gonadotropin. A high concentration of ET was found in the CL. The level of ET was significantly lower in the CL from TS-treated rats than that in TS-untreated rats (402.68 versus 575.60 pg/g wet weight, p less than 0.05). In contrast, the ET levels in plasma were by far lower than those in CL. These data indicate an inhibitory effect of TS on ET, an intraovarian peptide, production or accumulation in the CL.

Tokishakuyakusan stimulates cyclic adenosine 3',5'-monophosphate accumulation and progestin production by corpora lutea.

The effect of Tokishakuyakusan (TS) on rat corpora lutea was examined in vivo and in vitro. In an in vivo study, TS stimulated cyclic adenosine 3',5'-monophosphate (cyclic AMP) accumulation and steroidogenesis by corpora lutea, induced by PMS and hCG. In an incubation study, TS increased cyclic AMP accumulation and progesterone secretion. These results suggest that TS stimulates the corpora lutea to produce progestins via the mediation of cyclic AMP.

Hachimijiogan and Tokishakuyakusan enhance deoxyribonucleic acid alpha-nucleotidyltransferase activity in relationship to DNA synthesis by ovarian follicles via a cyclic AMP system.

The ovaries resected from twenty-nine day old female rats primed with 10 IU of PMS for 48 hours were incubated for 120 minutes with 2-20 micrograms/ml of Hachimijiogan (HZ) and Tokishakuyakusan (TS). The animals in another group were injected intravenously with 20 micrograms of HZ and TS 48 hours after PMS injection, and 20 minutes later decapitated. Deoxyribonucleic acid (DNA) alpha-nucleotidyltransferase activity and cyclic AMP accumulation in ovarian tissue were significantly increased by HZ and TS. These results suggest that HZ and TS enhance DNA alpha-nucleotidyltransferase activity via cyclic AMP by ovarian follicles.

Effects of Tokishakuyakusan and its components on rat corpus luteum function in vitro.

27-day-old female rats received 20 IU PMS and 56 hours later, 40 IU hCG. Seven days after hCG treatment, the resected ovaries were perifused with NIH-LH, Tokishakuyakusan (TJ-23) or its components. LH and TJ-23 increased progesterone in media and LH was more effective than TJ-23. Hoelen, Peony root, Alisma rhizome and Japanese angelica root also augmented progesterone. 17 alpha-hydroxyprogesterone and 20 alpha-hydroxyprogesterone were not stimulated by TJ-23. These results suggest that TJ-23 stimulates corpora lutea to secrete progesterone without direct luteotropic or luteolytic effect and that the effect may be attributable to Hoelen, Peony root, Alisma rhizome or Japanese angelica root.

Blended effects of herbal components of tokishakuyakusan on rat corpus luteum function in vivo.

The effect of herbal components of Tokishakuyakusan on progestin levels in serum and ovarian tissue from rats treated with PMS and hCG was examined in vivo. Hoelen + peony root + Japanese angelica root increased progesterone/20 alpha-OHP ratio in serum, and hoelen or peony root also increased the ratio in ovarian tissue, while atractylodes lanceae rhizome or hoelen + atractylodes lanceae rhizome decreased the ratio in serum and ovarian tissue. These data suggested that hoelen or peony root has a luteotropic effect but that atractylodes lanceae rhizome develops luteolysis. Furthermore, the data indicated a blended effect of herbal components of Tokishakuyakusan on the corpus luteum.

Hachimijiogan produces testosterone in adult rat testes.

The effect of Hachimijiogan (HZ) and Keishibukuryogan (KB) on the steroid production in rats was examined in vivo and in vitro. In an in vivo study, HZ stimulated the testes from ten-week old male rats to produce testosterone, whereas KB decreased the tissue testosterone concentrations. The delta 4-androstenedione and estradiol-17 beta (E2) showed no significant changes. In an incubation study, HZ also stimulated the testosterone production. The data suggested that HZ produces testosterone in rat testes. The role of KB is questionable.

Hachimijiogan changes serum hormonal circumstance and improves spermatogenesis in oligozoospermic men.

28 men with oligozoospermia and 7 with azoospermia received 7.5 g of Hachimijiogan, daily for 8 to 28 weeks. Examinations of semen in oligozoospermic men after treatment revealed a remarkable increase in number (78%) and moderate increase in sperm motility (53%) and in volume (56%), and fertility index was also remarkably improved, whereas no improvement was found in azoospermic men. Furthermore, serum estradiol-17 beta level was significantly increased (P less than 0.005) after treatment, while LH, FSH, prolactin and testosterone levels in sera showed no significant change. These results suggest that Hachimijiogan increases serum estradiol-17 beta levels and improves spermatogenesis in oligozoospermic men.