Acotiamide Hydrochloride, a Therapeutic Agent for Functional Dyspepsia, Enhances Acetylcholine-induced Contraction via Inhibition of Acetylcholinesterase Activity in Circular Muscle Strips of Guinea Pig Stomach.

Acotiamide is a first-in-class prokinetic drug approved in Japan for the treatment of functional dyspepsia. Given that acotiamide enhances gastric motility in conscious dogs and rats, we assessed the in vitro effects of this drug on the contraction of guinea pig stomach strips and on acetylcholinesterase (AChE) activity in stomach homogenate following fundus removal. We also investigated the serotonin 5-HT4 receptor agonist mosapride, dopamine D2 receptor and AChE inhibitor itopride, and representative AChE inhibitor neostigmine. Acotiamide (0.3 and 1 µM) and itopride (1 and 3 µM) significantly enhanced the contraction of gastric body strips induced by electrical field stimulation (EFS), but mosapride (1 and 10 µM) did not. Acotiamide and itopride significantly enhanced the contraction of gastric body and antrum strips induced by acetylcholine (ACh), but not that induced by carbachol (CCh). Neostigmine also significantly enhanced the contraction of gastric body strips induced by ACh, but not that by CCh. In contrast, mosapride failed to enhance contractions induced by either ACh or CCh in gastric antrum strips. Acotiamide exerted mixed inhibition of AChE, and the percentage inhibition of acotiamide (100 µM) against AChE activity was markedly reduced after the reaction mixture was dialyzed. In contrast, itopride exerted noncompetitive inhibition on AChE activity. These results indicate that acotiamide enhances ACh-dependent contraction in gastric strips of guinea pigs via the inhibition of AChE activity, and that it exerts mixed and reversible inhibition of AChE derived from guinea pig stomach.

Anomalous vertebral and posterior communicating arteries as a risk factor in instrumentation of the posterior cervical spine.

We investigated the incidence of anomalies in the vertebral arteries and Circle of Willis with three-dimensional CT angiography in 55 consecutive patients who had undergone an instrumented posterior fusion of the cervical spine. We recorded any peri-operative and post-operative complications. The frequency of congenital anomalies was 30.9%, abnormal vertebral artery blood flow was 58.2% and vertebral artery dominance 40%. The posterior communicating artery was occluded on one side in 41.8% of patients and bilaterally in 38.2%. Variations in the vertebral arteries and Circle of Willis were not significantly related to the presence or absence of posterior communicating arteries. Importantly, 18.2% of patients showed characteristic variations in the Circle of Willis with unilateral vertebral artery stenosis or a dominant vertebral artery, indicating that injury may cause lethal complications. One patient had post-operative cerebellar symptoms due to intra-operative injury of the vertebral artery, and one underwent a different surgical procedure because of insufficient collateral circulation. Pre-operative assessment of the vertebral arteries and Circle of Willis is essential if a posterior spinal fusion with instrumentation is to be carried out safely.

Acotiamide, a new orally active acetylcholinesterase inhibitor, stimulates gastrointestinal motor activity in conscious dogs.

UNLABELLED: BACKGROUND Acotiamide hydrochloride (acotiamide), a novel selective acetylcholinesterase (AChE) inhibitor, has proven significantly effective in treating functional dyspepsia (FD) in clinical trials, particularly in alleviating meal-related symptoms. In the present study, we examined the gastrointestinal prokinetic effects of acotiamide administered orally or intraduodenally in conscious dogs and investigated in vivo and ex vivo anti-AChE activity of acotiamide to clarify its mechanism of prokinetic action. METHODS: Gastrointestinal motility was measured in conscious dogs with chronically implanted force transducers. KEY RESULTS: Oral administration of acotiamide stimulated postprandial gastroduodenal and colonic motor activities. Measurement of gastrointestinal motility showed that acotiamide, like itopride and mosapride, enhanced gastric antral motility. Further, acotiamide markedly improved clonidine (an α(2) -adrenoceptor agonist)-induced hypomotility in a dog model of gastric motor dysfunction. The postprandial gastric antral motility enhanced by acotiamide was completely abolished on treatment with the muscarinic receptor antagonist atropine. Results of an in vivo experiment on anti-AChE activity showed clearly increased acetylcholine-induced gastric motility on intraduodenal administration of acotiamide, just as observed with the AChE inhibitor neostigmine. Further, in ex vivo experiment, intraduodenal administration of acotiamide significantly inhibited AChE activity in canine gastric antrum. CONCLUSIONS & INFERENCES: Our findings revealed that acotiamide administered through the alimentary tract exerts gastroprokinetic action via cholinergic pathways by inhibiting AChE activity. These results may also confirm the mechanism of action in clinical efficacy of acotiamide on FD.

Biodegradable Nano-aggregates of Star-Shaped 8-arm PEG-PLLA Block Co-polymers for Encapsulation of Water-Soluble Macromolecules.

A series of amphiphilic 8-arm PEG-b-PLLA co-polymers with star-shaped structure was synthesized through ring-opening polymerization of L-lactide (L-LA) in the presence of 8-arm PEG as a macroinitiator by varying feeding molar rations of L-LA to 8-arm PEG. 8-arm PEG-b-PLLA co-polymers having certain PEG content and PEG length were found to self-assemble into nano-aggregates in aqueous solutions. The size and the morphology of the nano-aggregates were investigated by dynamic light scattering and (1)H-NMR in CDCl3 and D2O. The results indicate that the average diameter was ca. 150 nm, the surface of the nano-aggregates was covered by PEG chains and the PLLA cores formed by hydrophobic interaction are located inside of the nano-aggregates. FITC-dextran molecules, as model for water-soluble macromolecular drugs, were successfully encapsulated into 8-arm PEG-b-PLLA nano-aggregates by simple addition of FITC-dextran to the aqueous phase during the self-assembly process. This result suggests that the nanoaggregates have a vesicle-like morphology. The nano-aggregates dissociated gradually in the order of weeks in PBS (pH 7.4, ionic strength 140 mM) at 37°C. Thus, the novel nano-aggregates of 8-arm PEG-b-PLLA can be expected to have advantages, such as long circulation times, as drug carriers which show sustained release of loaded macromolecular drugs such as antibodies and DNA vaccines in the blood stream.

Cutaneous metastasis of pancreatic carcinoma as an initial symptom in the lower extremity with obstructive lymphedema treated by physiotherapy and lymphaticovenous shunt: a case report, review, and pathophysiological implications.

Cutaneous metastasis from pancreatic cancer is relatively rare as an initial symptom, and it is generally localized on the periumbilical area that is known as Sister Mary Joseph's nodule. We report a rare case of a 49-year-old female who developed cutaneous metastasis of pancreatic cancer as an initial symptom. The patient was referred to our department for treatment of lymphedema due to surgical treatment of cervical cancer and underwent combined physiotherapy and, 2 months later, a lymph venous anastomosis (LVA) for treatment of the lymphedema. Two months after the operation, she developed erythema on her right leg which spread from the leg to the groin in series. This pattern corresponded to the direction of lymph drainage, which may have been enhanced by the conservative physiotherapy and LVA treatments. These facts suggest a possible relationship between cutaneous metastasis of carcinoma and treatment for lymphedema. Alternatively, the lymphedematous limb may be a privileged site for cancer growth, and metastatic seeding could have taken place from pre-existing hematogenous spread at the time of operation.

Genotyping of congenital adrenal hyperplasia due to 21-hydroxylase deficiency presenting as male infertility: case report and literature review.

We describe here two infertile male patients who were referred to our hospital with azoospermia at the ages of 33 and 30 years, respectively. Hormonal examinations led to a diagnosis of congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency in both patients. Genotyping revealed that the patients had a homozygous I172N and a heterozygous compound I172N/IVS2-13A/C>G mutation, respectively. Glucocorticoid replacement therapy succeeded in improving the seminal status of one patient, but not the other. For the latter patient and his wife, a pregnancy was achieved by testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) following genetic counseling. It is important to investigate genotyping and to classify patients on the basis of genotypic information in order to arrive at better treatment strategies for male infertility; especially in counseling of TESE-ICSI.

Possible role of autoantibodies against nephrin in an experimental model of chronic graft-versus-host disease.

Nephrin, a product of the NPHS1 gene, is a component of the slit diaphragms that are found between glomerular foot processes and is a crucial element for glomerular filtration barrier. Recently, nephrin has been focused in a number of studies of proteinuria development including various types of acquired glomerular diseases including minimal change nephrotic syndrome and membranous nephropathy. However, the precise role of nephrin in such acquired glomerular diseases is still unknown. To analyse the role of nephrin further, two kinds of anti-nephrin antibodies were raised in the rabbits and applied to an experimental mouse model of chronic graft-versus-host disease, in which (C57BL/10 x DBA/2) F1 mice developed clinically apparent severe proteinuria with significant glomerular lesions 7 weeks after parental DBA/2 cell transfer. Antibody-sandwich ELISA detected anti-nephrin antibodies during week 2 to week 6, with the peak at week 2 or week 4. Colocalization of nephrin and IgG on week 4, week 6, and week 8 was revealed by confocal microscopic analysis, suggesting that in situ immune complex formation with nephrin in glomerular lesion. Taken together, it seems to be suggested nephrin and its autoantibody have a certain role in the development of glomerular lesion in our model mice.

The deficiency of immunoregulatory receptor PD-1 causes mild osteopetrosis.

Recently, the involvement of immune responses in metabolic bone disease and/or local bone destruction has received much attention. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), a member of the immunoglobulin (Ig) superfamily, negatively regulates T cell activation. The deficiency of CTLA-4 induces profound osteopenia with an increase in osteoclastogenesis, suggesting the important role of activated T cells in osteoclastogenesis. Programmed death-1 (PD-1) is the newly identified immunoregulatory receptor, which also belongs to the Ig superfamily. Both CTLA-4 and PD-1 are induced on activated T cells, however, there are no reports linking PD-1 with osteoclasts. In the present study, we have examined the bone phenotype in PD-1-deficient mice PD-1-/- and the role of PD-1 in osteoclastogenesis and osteoclast function. Both trabecular and cortical bone mineral densities of tibia were significantly increased, as observed in peripheral quantitative computed tomography (pQCT), at 12 weeks of age in PD-1-/- mice. Histomorphometric analysis of the PD-1-/- mice and the age-matched controls at 12 weeks of age showed a 2-fold increase in bone volume (BV/TV) with a 55% decrease in osteoclast number (N.Oc/BS). Bone formation indices were similar in both groups. The number of soluble receptor activator of nuclear factor kappaB ligand (sRANKL)-induced osteoclast-like cells (OCLs) derived from the PD-1-deficient splenocytes was significantly decreased (by 25%). On the other hand, PD-1 deficiency did not affect the bone-resorbing activity of mature osteoclasts. Our results suggest that PD-1 deficiency reduces osteoclastogenesis resulting in an osteopetrotic phenotype. Identical members of the Ig superfamily, CTLA-4 and PD-1, which negatively regulate immune responses, may differentially affect osteoclastogenesis and bone remodeling.

Chemical induction of HO-1 suppresses lupus nephritis by reducing local iNOS expression and synthesis of anti-dsDNA antibody.

There is accumulating evidence that haem oxygenase (HO)-1 plays a protective role in various disorders. The beneficial efficacy of HO-1 induction therapy has been shown in renal diseases such as glomerulonephritis, interstitial nephritis and drug induced nephrotoxicity. However, involvement of HO-1 in the development of autoimmune renal diseases remains uncertain. To assess the clinical efficacy of HO-1 induction therapy for lupus glomerulonephritis, MRL/lpr mice were intraperitoneally injected with 100 micromol/kg hemin, a potent HO-1 inducer, or PBS as controls, once a week from 6 weeks of age to 21-24 weeks-old. We found that treatment with hemin led to a significant reduction of proteinuria and remarkable amelioration of glomerular lesions accompanied by decreased immune depositions. In addition, the circulating IgG anti-double-stranded DNA antibody level was significantly decreased in hemin treated mice when compared with controls. A single intraperitoneal injection with hemin resulted in reduction of inducible nitric oxide synthase expression in the kidney and spleen, and serum interferon-gamma level. Our results suggest that HO-1 induction therapy ameliorates lupus nephritis by suppressing nitric oxide (NO) dependent inflammatory responses and attenuating production of pathogenic autoantibodies.

Optimization of fermentation reaction conditions for preparation of PATE enzyme by Erwinia carotovora IFO3830 by TFCCRD method.

The fermentation conditions for preparation of polygalacturonic acid transeliminase (PATE) enzyme by Erwinia carotovora IFO3830 were optimized for seed ratio, vibration rate, and temperature by the TFCCRD method. The results indicated that the optimum fermentation conditions for E. carotovora IFO 3830 were that seed ratio, vibration rate, and temperature were 5% v/v, 113 min(-1), and 29 degrees C, respectively.

[Clinical outcome of potassium-titanyl-phosphate (KTP/532) laser vaporization prostatectomy for benign prostate hyperplasia].

OBJECTIVE: To study the feasibility of vaporization prostatectomy by potassium-titanyl-phosphate (KTP/532) laser in men with bladder outlet obstruction due to benign prostatic hyperplasia (BPH) and to evaluate their clinical and voiding outcome 2 weeks and 6 months after operation. PATIENTS AND METHODS: Laser vaporization prostatectomy with the KTP/532 laser at 20 W was performed in 21 BPH patients varying from 56 to 75 years of age. All cases were performed under the spinal anesthesia. The laser was produced by MODEL SL20/50 and ADD/Stat side-firing fibers were used. A urethral catheter was inserted post-operatively and was removed within 6 days after operation. The hemoglobin value was examined on the first day after operation and uroflowmetry was executed 2 weeks and 6 months after operation with the examination of the residual urine volume. IPSS and QOL score were recorded by patients 2 weeks and 6 months after operation. RESULTS: The prostate volumes ranged from 22 to 52 mL (mean 37.3 +/- 10.4). None of the 21 patients had any significant blood loss or any fluid absorption. The voiding parameters improved: mean peak flow rate increased from 7.1 to 16.5 mL/s 2 weeks after operation and to 15.2 mL/s 6 months after operation and residual volume decreased from 172 to 45 mL 2 weeks after operation and to 43 mL 6 months after operation. IPSS and QOL score improved significantly in Wilcoxon signed-ranks test. The ratio of excellent and good in efficacy criteria of Kawabe group was 71.4% 2 weeks after operation and that was 71.4% 6 months after operation. Mean catheterization term was 4.5 days. Only one patient showed transient urinary retention when the catheter was removed the third postoperative day. None had incontinence postoperatively. No other significant complications were recognized. The prostate volume was more than 50 mL in 3 cases and the postoperative 6 months outcomes of the 3 cases were all poor in their symptoms in the efficacy criteria of Kawabe group. CONCLUSIONS: KTP laser vaporization prostatectomy is safe and effective for obstructive prostates up to 50 mL in volume and produces good results 2 weeks post operatively that are sustained for 6 months after operation.

[Giant epidermal cyst in the perineum extending into the pelvic space in the patient with polycystic kidney disease: a case report].

A 75-year-old man with autosomal dominant polycystic kidney disease was admitted to our hospital with the chief complaint of a giant mass in the left side of the perineum that had gradually developed in size during the previous 20 years. Palpation revealed a neonatal head-sized cystic mass with no tenderness. Magnetic resonance imaging (MRI) demonstrated a well-circumscribed homogeneous mass extended to the dorsal side of the bladder. It was excised surgically without any injury of rectum or urinary tract. The resected specimen was 23 x 18 x 12 cm in size and 1.2 kg in weight. The pathological diagnosis was epidermal cyst. Cases with an epidermal cyst extending into the pelvic space are extremely rare with few cases having been reported in the world.

Optimization of fermentation conditions for preparation of polygalacturonic acid transeliminase by Erwinia carotovora IFO3830.

This paper is concerned with optimization of the fermentation conditions for preparation of alkaline pectin lyase (poly-galacturonic acid trans-eliminase, PATE) by Erwinia carotovora IFO3830 (E.C.IFO3830). The orthogonal matrix method was adopted as the experimental design method for media. The effects of media composition on the growth rate of E.C.IFO3830 are in the order of pectin > KH(2)PO(4)/Na(2)HPO(4) > MgSO(4) > (NH(4))(2)SO(4) > glucose > L-glutamate sodium, and those on PATE activity are in the order of glucose > pectin > L-glutamate sodium > KH(2)PO(4)/Na(2)HPO(4) > MgSO(4) > (NH(4))(2)SO(4). The strain of E.C.IFO3830 grows quickly and easily near an initial pH of 7.0, and especially at the alkaline range of pH 7.0-8.0. The growth rate profiles indicate that E.C.IFO3830 grows very quickly; its generation time t(1/2) is about 0.2 h. The appropriate fermentation period is about 10-20 h for a high level productivity of biomass and 25-35 h for high productivity of PATE enzyme.

Need for sling surgery in patients with large cystoceles and masked stress urinary incontinence.

BACKGROUND: We studied the need for sling surgery in patients who suffered from large cystoceles and masked stress urinary incontinence. METHODS: Twenty patients who had large cystoceles but neither evidence nor history of stress incontinence were enrolled in this study. The cystocele was reduced using a reducing device. Masked urinary incontinence was identified by a 60 m pad weighing test and a stress test. The cystocele was reduced using a pessary ring in 14 patients, or a vaginal pack formed of two rolls of ordinary 28 x 28 cm gauze in six patients. Ten of 20 patients were diagnosed with masked stress urinary incontinence and were treated with anterior colporrhaphy and a suburethral sling procedure. The other 10 patients were continent after use of a cystocele reducing device and were treated with anterior colporrhaphy alone. Average follow-up periods of the patients with or without masked stress urinary incontinence were 51.2 months (range, 24.0-72.0 months) or 57.6 months (range, 27.0-70.0 months), respectively. RESULTS: One of the 10 patients diagnosed with masked stress incontinence had mild stress urinary incontinence postoperatively. None of the 10 continent patients had stress incontinence after anterior colporrhaphy alone. CONCLUSIONS: Reducing devices of protruding cystocele were clinically useful in the detection of masked stress incontinence. Sling surgery was effective to prevent emerging stress urinary incontinence for patients who suffered from cystocele and masked stress incontinence.

[Retroperitoneal ganglioneuroma: a case report].

A primary retroperitoneal ganglioneuroma was found incidentally in a 44-year-old man. Computed tomography revealed a homogeneous mass just above the right kidney. T1-weighted magnetic resonance imaging demonstrated the homogeneous mass, but T2-weighted imaging revealed a heterogeneous mass with a markedly high intensity area. The tumor was resected through a transabdominal approach. The resected specimen measured 11 x 9 x 4.5 cm and weighted 270 g. Histological examination showed that the lesion was a ganglioneuroma composed of mature ganglion cells and nerve fibers.

Polytetrafluoroethylene patch sling for type 2 or type 3 stress urinary incontinence.

BACKGROUND: Surgical success rates and complications of anti-incontinence surgery were evaluated according to the type of stress incontinence and the type of surgery. METHODS: From 1989 to 1998, we treated 137 women for stress urinary incontinence with anti-incontinence surgery. Of the 137 patients, 110 had type 2 stress urinary incontinence and 27 had type 3 stress urinary incontinence. Of 110 patients with type 2 stress urinary incontinence, 57 underwent pubovaginal sling procedure and 53 were treated with the Gittes procedure. All of the patients with type 3 stress urinary incontinence underwent the pubovaginal sling procedure. RESULTS: Cure rates with the pubovaginal sling procedure were 82% in type 2 incontinence and 70% in type 3 incontinence. Cure rates with the Gittes procedure were 56%. The occurrence of de novo urge symptom and pelvic pain was low and bore no relation to either operative method or type of stress incontinence. CONCLUSIONS: The pubovaginal sling procedure using the polytetrafluoroethylene patch was effective for type 2 or 3 stress urinary incontinence.

[A case of giant hemorrhagic adrenal pseudocyst with infection].

A 51-year-old woman was admitted with general fatigue, high fever and left upper abdominal pain. Abdominal computed tomography revealed a left adrenal mass, 15 cm in diameter with regular margins, the contents of which were not enhanced with contrast medium. The mass had heterogeneous echogenicity on the abdominal ultrasonograph. Serum hormonal levels were almost normal. Despite treatment with antibiotics, the high fever persisted. Based on these findings, the differential diagnosis was adrenal abscess or a nonfunctional adrenal tumor. Transabdominal left adrenalectomy was performed. Pathological examination revealed a hemorrhagic adrenal pseudocyst with infection. The patient's condition improved soon after the operation.

Splenectomy may improve the glomerulopathy of type II mixed cryoglobulinemia.

Many patients with type II mixed cryoglobulinemia have been shown to be infected with hapatitis C virus (HCV). Therefore, interferon-alfa has become the first choice of treatment for patients with HCV-associated cryoglobulinemia. However, the disease often relapses after the discontinuation of interferon therapy. The long-term effect of interferon therapy is controversial. Therefore, a more effective therapy needs to be developed. A 62-year-old Japanese woman was admitted to our hospital for the examination of abnormal liver function tests, severe edema, and purpura in her lower extremities. Glomerulopathy secondary to HCV-related cryoglobulinemia was suspected. Her serum creatinine was increased to 2.1 mg/dL. Interferon therapy was considered initially. However, because of pancytopenia caused by liver cirrhosis and splenomegaly, splenectomy was performed in February 1997, before the start of interferon therapy. Renal biopsy specimen taken at the time of the splenectomy showed typical cryoglobulinemic glomerulonephritis. Gradually, after surgery, the patient's thrombocytopenia and anemia improved, her proteinuria and hematuria were decreased, her cryocrit dropped from 15% to 5%, the Ccr increased from 21.1 mL/min to 48.8 mL/min, and the purpura in her lower extremities disappeared. A repeat renal biopsy performed in May 1998 showed marked histological improvement. Splenectomy is not widely accepted as a treatment for cryoglobulinemia. Our case suggests the possibility that the monoclonal-IgM component of the type II cryoglobulin may be formed in the spleen. In conclusion, splenectomy may be an effective therapy for cryoglobulinemia in patients with HCV-positive liver cirrhosis and pancytopenia secondary to splenomegaly.

[Crossed fused kidney with ectopic ureter: a case report].

A 5-year-old girl with a rare anomaly of a single system ectopic ureter draining into the mesonephric duct cyst and ipsilateral crossed fused kidney is presented. Both ureters were catheterized and three dimensional (3-D) images of the urinary tract were reconstructed with helical computed tomographic (CT) data. A mesonephric duct cyst, the right ureter draining into the cyst and an ectopic orifice were well depicted with these images, especially with inferior projection images. 3-D display of helical CT data with a retrograde contrast study is a promising method of obtaining anatomical detail of an ectopic ureter.