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Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used for the treatment of inflammatory pain. It is well established that NSAIDs exert their analgesic effects by inhibiting cyclooxygenase to prevent the production of prostaglandins; however, several NSAIDs including diclofenac also modulate other ion channels expressed in nociceptive neurons. In this study, we investigated the pH-dependent effects of diclofenac on tetrodotoxin-resistant (TTX-R) Na + channels in rat trigeminal sensory neurons by using the whole-cell patch clamp technique. Diclofenac decreased the peak amplitude of TTX-R Na + currents (I Na ) in a concentration dependent manner. While diclofenac had little effect on the voltage-activation relationship, it significantly shifted the steady-state fast inactivation relationship toward hyperpolarized potentials. Diclofenac increased the extent of use-dependent inhibition of TTX-R Na + currents. Diclofenac also significantly accelerated the development of inactivation and retarded the recovery from inactivation of TTX-R Na + channels. The effects of diclofenac on TTX-R Na + channels were stronger at pH 6.0 than at pH 7.4 for most of the parameters tested. Considering that the extracellular pH falls in inflamed tissues, and that TTX-R Na + channels expressed on nociceptive neurons are implicated in the prostaglandin-mediated development and maintenance of inflammatory hyperalgesia, our findings could provide an additional analgesic effect of diclofenac under acidic pH conditions.

A cDNA (PyARP4) containing an open reading frame for a protein of 573 amino acids was identified in the marine red alga Porphyra yezoensis. The conceptual PyARP4 protein exhibits significant similarity to actin-related protein (ARP) 4 in the terrestrial plant Arabidopsis. Comparison of the deduced amino acid sequence showed moderate sequence identity (30%) to a conventional actin in P. yezoensis, as seen in comparisons between ARP and conventional actins of other organisms. A putative bipartite nuclear localization signal and an actin motif were found within the PyARP4 amino acid sequence. In a phylogenetic analysis, the PyARP4 was found to cluster with the ARP4 of other organisms. The expression level of PyARP4 did not change significantly among four developmental stages of life cycle and was lower than that of a conventional actin. This cDNA therefore may serve as a useful internal standard in gene expression analyses of differentially expressed genes in P. yezoensis.

MAMLD1 is a causative gene for disorders of sex development. Several MAMLD1 mutations have been shown to cause hypospadias by generating dysfunctional proteins and/or unstable mRNAs. Here, we identified an intronic mutation of MAMLD1 (g.IVS4−2A>G) in 1 of 180 hypospadias patients. RT-PCR of the patient's skin sample showed normal expression of full-length MAMLD1 and markedly reduced expression of a known splice variant lacking exon 4. A hitherto unreported splice variant that lacks exon 5 was similarly identified in samples of the patient and control individuals. The full-length transcript of the patient contained mutant mRNA lacking the first 10 nucleotides of exon 5 (c.1822_1831delACTCATGTAG, p.K609fsX1070). In vitro assays using cells expressing the full-length wild-type and mutant proteins revealed reduced expression of the mutant. The expression of the wild-type and mutant MAMLD1 showed parallel changes upon treatment with a proteasome inhibitor and a translation inhibitor. The mutant-expressing cells exerted low transactivation activity for the Hes3 promoter, which reflected limited expression of the mutant protein. These results imply that the pathogenic events resulting from MAMLD1 mutations include splice errors. Furthermore, this study raises the possibility of translation failure of MAMLD1 mutants, which deserves further investigation.

The Japanese Schizophrenia Sib-Pair Linkage Group (JSSLG) is a multisite collaborative study group that was organized to create a national resource for affected sib pair (ASP) studies of schizophrenia in Japan. We used a high-density single-nucleotide-polymorphism (SNP) genotyping assay, the Illumina BeadArray linkage mapping panel (version 4) comprising 5,861 SNPs, to perform a genomewide linkage analysis of JSSLG samples comprising 236 Japanese families with 268 nonindependent ASPs with schizophrenia. All subjects were Japanese. Among these families, 122 families comprised the same subjects analyzed with short tandem repeat markers. All the probands and their siblings, with the exception of seven siblings with schizoaffective disorder, had schizophrenia. After excluding SNPs with high linkage disequilibrium, we found significant evidence of linkage of schizophrenia to chromosome 1p21.2-1p13.2 (LOD=3.39) and suggestive evidence of linkage to 14q11.2 (LOD=2.87), 14q11.2-q13.2 (LOD=2.33), and 20p12.1-p11.2 (LOD=2.33). Although linkage to these regions has received little attention, these regions are included in or partially overlap the 10 regions reported by Lewis et al. that passed the two aggregate criteria of a meta-analysis. Results of the present study--which, to our knowledge, is the first genomewide analysis of schizophrenia in ASPs of a single Asian ethnicity that is comparable to the analyses done of ASPs of European descent--indicate the existence of schizophrenia susceptibility loci that are common to different ethnic groups but that likely have different ethnicity-specific effects.

Upon activation of cell surface receptors coupled to the Gq subclass of G proteins, phospholipase C (PLC) beta hydrolyses membrane phospholipid to yield a pair of second messengers, inositol 1,4,5-trisphosphate (IP3) and 1,2-diacylglycerol. PLCbeta4 has been characterized as the isoform enriched in cerebellar Purkinje cells (PCs) and the retina and involved in motor and visual functions. Here we examined cellular and subcellular distributions of PLCbeta4 in adult mouse brains. Immunohistochemistry showed that high levels of PLCbeta4 were detected in the somatodendritic domain of neuronal populations expressing the metabotropic glutamate receptor (mGluR) type 1alpha, including olfactory periglomerular cells, neurons in the bed nucleus anterior commissure, thalamus, substantia nigra, inferior olive, and unipolar brush cells and PCs in the cerebellum. Low to moderate levels were detected in many other mGluR1alpha-positive neurons and in a few mGluR1alpha-negative neurons. In PCs, immunogold electron microscopy localized PLCbeta4 to the perisynapse, at which mGluR1alpha is concentrated, and to the smooth endoplasmic reticulum in dendrites and spines, an intracellular Ca2+ store gated by IP3 receptors. In the cerebellum, immunoblot demonstrated its concentrated distribution in the post-synaptic density and microsomal fractions, where mGluR1alpha and type 1 IP3 receptor were also greatly enriched. Furthermore, PLCbeta4 formed coimmunoprecipitable complexes with mGluR1alpha, type 1 IP3 receptor and Homer 1. These results suggest that PLCbeta4 is preferentially localized in the perisynapse and smooth endoplasmic reticulum as a component of the physically linked phosphoinositide signaling complex. This close molecular relationship might provide PLCbeta4 with a high-fidelity effector function to mediate various neuronal responses under physiological and pathophysiological conditions.

GABAergic afferent inputs are thought to play an important role in the control of the firing pattern of substantia nigra pars compacta (SNc) dopaminergic neurons. We report here the actions of presynaptic kainite (KA) receptors in GABAergic transmission of rat SNc dopaminergic neurons. In mechanically dissociated rat SNc dopaminergic neurons attached with native presynaptic nerve terminals, GABAergic miniature inhibitory postsynaptic currents (mIPSCs) were recorded by use of conventional whole cell patch recording mode. In the voltage-clamp condition, KA (3 μM) significantly increased GABAergic mIPSC frequency without affecting the current amplitude. This facilitatory effect of KA was not affected in the presence of 20 μM GYKI52466, a selective AMPA receptor antagonist, but was completely inhibited in the presence of 20 μM CNQX, an AMPA/KA receptor antagonist. Presynaptic KA receptors on GABAergic terminals were mainly permeable to Na+ but impermeable to Ca2+ because KA-induced facilitation of mIPSC frequency was completely suppressed in either Na+-free or Ca2+-free external solutions, and in the presence of 200 μM Cd2+, a general voltage-dependent Ca2+ channel blocker. In the slice preparation, KA increased GABAergic spontaneous mIPSC frequency, but significantly suppressed evoked IPSC (eIPSC) amplitude. However, this inhibitory action on eIPSCs was reversed by 10 μM CGP55845 , a selective GABAB receptor antagonist, implicating the possible involvement of GABAB autoreceptors in KA-induced modulation of GABAergic transmission. Thus presynaptic KA receptors on GABAergic nerve terminals synapsing onto SNc neurons may play functional roles contributing the fine control of neuronal excitability and firing pattern of SNc.

A 73-year-old man presented with gross hematuria. Ultrasonography and computerized tomography showed small bladder tumors and a left renal mass protruding to renal pelvis. Transurethral resection of bladder tumor and ureteroscopic tumor biopsy were performed, and pathological examinations revealed transitional cell carcinoma in the bladder and renal cell carcinoma in the kidney. He underwent left radical nephrectomy. A 4-month postoperative cystoscopy revealed a solitaly non-papillary tumor in the bladder. Transurethral resection was performed and pathological diagnosis was metastasis from renal cell carcinoma. At that time, multiple metastases to ureteral stump and lung were found. He had undergone palliative treatment because of his poor general condition until he died 26 months postoperatively. Care should be taken for management of ureteral stump when diagnostic ureteroscopy was done for renal cell carcinoma invading the renal pelvis.

Cell growth of anaerobic protozoan Tritrichomonas foetus was analyzed. This protozoan usually proliferates in extremely high density, but protozoan parasites were dispersed uniformly in F-bouillon medium and cell division stopped temporarily. However, nuclear fission continued and giant polynucleated cells formed. Later, cell division resumed and cells returned to normal form. In conditioned medium, cytokinesis of the dispersed parasites did not stop. Results indicated that T. foetus cells secreted an extra-cellular factor that influenced cytokinesis.

We retrospectively assessed the incidence of and risk factors for febrile urinary tract infection in children during active surveillance after early discontinuation of antibiotic prophylaxis.We retrospectively evaluated 9 females and 61 uncircumcised males diagnosed with primary vesicoureteral reflux before age 1 year who had persistent reflux on followup voiding cystourethrogram and were subsequently followed under active surveillance without continuous antibiotic prophylaxis. Patients with secondary vesicoureteral reflux or associated urological abnormality were excluded. Clinical outcomes, including incidence of febrile urinary tract infection and new scar formation, were evaluated. Risk factors for febrile urinary tract infection were also analyzed.Mean age at stopping continuous antibiotic prophylaxis was 21 months, and mean followup was 61 months. During active surveillance 21 patients had febrile urinary tract infection, and the 5-year infection-free rate under active surveillance was 67.5%. One or 2 foci of minimal new scarring developed in 4 of 16 patients who underwent followup dimercapto-succinic acid scan after febrile urinary tract infection. On multivariate analysis dilated vesicoureteral reflux on followup voiding cystourethrogram was the only significant risk factor for febrile urinary tract infection.This study revealed that about two-thirds of patients with persistent vesicoureteral reflux were free of febrile urinary tract infection during 5 years of active surveillance. Those with dilated vesicoureteral reflux on followup voiding cystourethrogram are at significantly greater risk for febrile urinary tract infection. Accordingly active surveillance, especially in patients with nondilated vesicoureteral reflux on followup voiding cystourethrogram, seems to be a safe option even in children who have not yet been toilet trained.

Under pathological conditions including inflammation, ischemia and incision, extracellular pH falls down as low as 5.4. Although some mediators play pivotal roles in the development and maintenance of inflammatory hyperalgesia by affecting the functional properties of tetrodotoxin-resistant (TTX-R) Na(+) channels, the roles of tissue acidosis in nociceptive transmission mediated by TTX-R Na(+) channels are largely unknown. In the present study, we have investigated the effect of acidic pH on TTX-R Na(+) currents (I(Na)) in small-sized sensory neurons isolated from rat trigeminal ganglia using a whole-cell patch clamp technique. Acidic pH decreased the peak amplitude of TTX-R I(Na) in a pH-dependent manner, but weak acid (≥pH 6.0) had a minor inhibitory effect on the TTX-R I(Na). Acidic pH also significantly shifted both the activation and steady-state fast inactivation relationships toward depolarized potentials. In addition, acidic pH had little effect on the use-dependent inhibition, and significantly retarded the development of inactivation and accelerated the recovery from inactivation of TTX-R Na(+) channels. The results suggest that weak acid (≥pH 6.0) makes TTX-R Na(+) channels to be suitable for the repetitive activation at depolarized membrane potentials. Given that both tissue acidosis and inflammatory mediators in inflamed or injured tissues act synergistically to promote nociceptive transmission by affecting the functional properties of TTX-R Na(+) channels, these channels would be, at least in part, a good target to treat inflammatory pain.

Acid-sensing ion channels (ASICs) are widely expressed in central as well as peripheral neurons. Here we have characterized the proton-induced currents in acutely isolated rat trigeminal mesencephalic nucleus (Vmes) neurons using a whole cell patch-clamp technique. In a voltage-clamp condition, the application of acid extracellular solution (≤ pH 6.5) induced the inward currents in a pH-dependent manner. The proton-induced currents disappeared in the Na(+)-free external solution, and were concentration-dependently blocked by amiloride, a general ASIC blocker. The reversal potential of proton-induced currents was similar to the theoretical Na(+) equilibrium potential, suggesting that the proton-induced currents are mainly mediated by the activation of ASICs, which are highly selective to Na(+). The modulation of proton-induced currents by divalent cations and the expression patterns of ASIC transcripts using by the multi-cell RT-PCR assay suggest that Vmes neurons express functional ASIC2a and ASIC1b subunits. In a current-clamp condition, acidic pH directly depolarized the membrane potential and generated a burst of action potentials at Vmes neurons, which innervate the masseter muscle spindles. Considering that cell bodies of Vmes neurons are located within the central nervous system, ASICs expressed on Vmes neurons, by sensing peripheral and/or central acidosis, might play pivotal roles in the transduction of proprioceptive information from the masseter muscles and periodontal ligaments.

Formation of the forespore membrane, which becomes the plasma membrane of spores, is an intriguing step in the sporulation of the fission yeast Schizosaccharomyces pombe. Here we report two novel proteins that localize to the forespore membrane.spo3+encodes a potential membrane protein, which was expressed only during sporulation. Green fluorescent protein (GFP) fusion revealed that Spo3 localized to the forespore membrane. The spo3 disruptant was viable and executed meiotic nuclear divisions as efficiently as the wild type but did not form spores. One of the spo3 alleles,spo3-KC51, was dose-dependently suppressed bypsy1+, which encodes a protein similar to mammalian syntaxin-1A, a component of the plasma membrane docking/fusion complex. psy1+was essential for vegetative growth, and its transcription was enhanced during sporulation. As expected, Psy1 localized to the plasma membrane during vegetative growth. Interestingly, Psy1 on the plasma membrane disappeared immediately after first meiotic division and relocalized to the forespore membrane as the second division initiated. In thespo3 null mutant, the forespore membrane was initiated but failed to develop a normal morphology. Electron microscopy revealed that membrane vesicles were accumulated in the cytoplasm of immaturespo3Δ asci. These results suggest that Spo3 is a key component of the forespore membrane and is essential for its assembly acting in collaboration with the syntaxin-like protein.

Ligand binding to neurotransmitter and hormone receptors which couple to the Gq subclass of GTP-binding protein leads to the activation of phospholipase Cbeta (PLCbeta) which hydrolyses phosphatidyl-inositol 4,5-bisphosphate, yielding a pair of second messengers, diacylglycerol and inositol 1,4,5-trisphosphate (IP3). The expression of PLCbeta1-4 mRNAs was comparatively examined by in situ hybridization in the mouse brain. In adults, PLCbeta1 mRNA was expressed predominantly in the telencephalon, including the cerebral cortex, hippocampus, amygdala, lateral septum and olfactory bulb, with little expression in most thalamic nuclei. PLCbeta2 mRNA was distributed in the white matter, suggesting its expression in non-neuronal cells, most likely oligodendrocytes. PLCbeta3 mRNA was specifically expressed in cerebellar Purkinje cells. The highest levels of PLCbeta4 mRNA were detected in Purkinje cells. High levels of PLCbeta4 mRNA were also found in the thalamus and medial septum, whereas weak signals were detected in most telencephalic regions, thus showing an expression pattern almost reciprocal to that of PLCbeta1 mRNA. During development, such characteristic regional expression of PLCbeta1 and PLCbeta4 were observed starting in late foetal stages, while specific expression of PLCbeta2 and PLCbeta3 appeared in early postnatal stages. We conclude that despite the existence of four PLCbeta isoforms, only one or two of them is expressed in individual neurons and glial cells. The distinct expression of PLCbetas provides a molecular basis for analysing the nature of the specific signal transduction pathway leading to the production of diacylglycerol and IP3 in distinct cell types and in different regions of the brain.

A 38-year-old female was transfered to our hospital due to hypotension. Echocardiography revealed diffuse severe hypokinesis. Circulatory support with intraaortic balloon pumping (IABP) and percutaneous cardiopulmonary support (PCPS) was initiated for fulminant myocarditis. Severe hypokinesis continued and motion of the valves was not recognized. PCPS was continued for 340 hours, IABP for 415 hours, artificial ventilation for 30 days, and continuous hemodiafiltration for acute renal failure for 40 days. The wall motion of the heart recovered completely as did renal function. The patient was discharged on the 189th day without neurologically consequences. Circulatory support was carried out safely and effectively in a patient with severe hypokinetic heart due to fulminant myocarditis.

An 18-year-old female was admitted to our ICU for dyspnea. She had bloody sputum, hypoxia and hypoproteinemia with proteinuria. Massive doses of methylprednisolone (750mg·day-1) were administered for 3 days. She recovered temporarily. Renal biopsy revealed crescentic glomerulonephritis and transbronchial lung biopsy revealed alveolar hemorrhage. The examination of autoantibodies was positive for perinuclear antineutrophil cytoplasmic autoantibodies (P-ANCA). Pulmonary and renal function deteriorated gradually despite the administration of prednisolone (60mg·day-1). Plasma exchange and hemodialysis were performed and massive doses of methylprednisolone (1g·day-1) were administered for 3 days. Pulmonary function recovered, but it was necessary to continue hemodialysis. Pulmonary function deteriorated again one month later. P-ANCA tested negative after plasma absorption, but pulmonary function grew worse as abnormalities in blood coagulation progressed. The patient died from hemorrhagic cerebral infarction. We believe this was a case of Wegener's vasculitis with P-ANCA. We administered of massive doses of methylprednisolone, and performed plasma exchange and plasma absorption, but these therapies proved to be ineffective. The levels of P-ANCA, white blood cells and C-reactive protein did not reflect the virulence of the disease in this patient. It is important to investigate the indices that reflect the activity of this disease and the choice of the appropriate therapy for the level of virulence.

During mammalian embryonic development, the primitive streak initiates the differentiation of pluripotent epiblast cells into germ layers. Pluripotency can be reacquired in committed somatic cells using a combination of a handful of transcription factors, such as OCT3/4, SOX2, KLF4 and c-MYC (hereafter referred to as OSKM), albeit with low efficiency. Here we show that during OSKM-induced reprogramming towards pluripotency in human cells, intermediate cells transiently show gene expression profiles resembling mesendoderm, which is a major component of the primitive streak. Based on these findings, we discover that forkhead box H1 (FOXH1), a transcription factor required for anterior primitive streak specification during early development, significantly enhances the reprogramming efficiency of human fibroblasts by promoting their maturation, including mesenchymal to epithelial transition and the activation of late pluripotency markers. These results demonstrate that during the reprogramming process, human somatic cells go through a transient state that resembles mesendoderm.

While sialylation plays important functions in the nervous system, the complexity of glycosylation pathways and limitations of genetic approaches preclude the efficient analysis of these functions in mammalian organisms. Drosophila has recently emerged as a promising model for studying neural sialylation. Drosophila sialyltransferase, DSiaT, was shown to be involved in the regulation of neural transmission. However, the sialylation pathway was not investigated in Drosophila beyond the DSiaT-mediated step. Here we focused on the function of Drosophila cytidine monophosphate-sialic acid synthetase (CSAS), the enzyme providing a sugar donor for DSiaT. Our results revealed that the expression of CSAS is tightly regulated and restricted to the CNS throughout development and in adult flies. We generated CSAS mutants and analyzed their phenotypes using behavioral and physiological approaches. Our experiments demonstrated that mutant phenotypes of CSAS are similar to those of DSiaT, including decreased longevity, temperature-induced paralysis, locomotor abnormalities, and defects of neural transmission at neuromuscular junctions. Genetic interactions between CSAS, DSiaT, and voltage-gated channel genes paralytic and seizure were consistent with the hypothesis that CSAS and DSiaT function within the same pathway regulating neural excitability. Intriguingly, these interactions also suggested that CSAS and DSiaT have some additional, independent functions. Moreover, unlike its mammalian counterparts that work in the nucleus, Drosophila CSAS was found to be a glycoprotein-bearing N-glycans and predominantly localized in vivo to the Golgi compartment. Our work provides the first systematic analysis of in vivo functions of a eukaryotic CSAS gene and sheds light on evolutionary relationships among metazoan CSAS proteins.

Histaminergic neurons within the tuberomammillary nucleus (TMN) play an important role in the regulation of sleep-wakefulness. Here, we report the adrenergic modulation of GABAergic transmission in rat TMN histaminergic neurons using a conventional whole-cell patch clamp technique. Norepinephrine (NE) reversibly decreased the amplitude of action potential-dependent GABAergic inhibitory post-synaptic currents (IPSCs) and increased the paired pulse ratio. The NE-induced inhibition of GABAergic IPSCs was mimicked by clonidine, a selective α2 adrenoceptor agonist. However, cirazoline and isoproterenol, nonselective α1 and β adrenoceptor agonists, respectively, had no effect on GABAergic IPSCs. The NE-induced inhibition of GABAergic IPSCs was significantly blocked by BRL44408, a selective α2A adrenoceptor antagonist, but not imiloxan or JP1302, a selective α2B and α2C adrenoceptor antagonists. The extent of NE-induced inhibition of GABAergic IPSCs was inversely proportional to the extracellular Ca(2+) concentration. Pharmacological agents affecting the activities of adenylyl cyclase or G-protein-coupled inwardly rectifying K(+) channels did not affect the NE-induced inhibition of GABAergic IPSCs. However, NE had no effect on the frequency and amplitude of GABAergic miniature IPSCs. These results suggest that NE acts on presynaptic α2A adrenoceptor to inhibit action potential-dependent GABA release via the inhibition of Ca(2+) influx from the extracellular space to GABAergic nerve terminals, and that this α2A adrenoceptor-mediated modulation of GABAergic transmission may be involved in regulating the excitability of TMN histaminergic neurons.

The periaqueductal gray (PAG) is involved in the central regulation of nociceptive transmission by affecting the descending inhibitory pathway. In the present study, we have addressed the functional role of presynaptic glycine receptors in spontaneous glutamatergic transmission. Spontaneous EPSCs (sEPSCs) were recorded in mechanically dissociated rat PAG neurons using a conventional whole-cell patch recording technique under voltage-clamp conditions. The application of glycine (100 µM) significantly increased the frequency of sEPSCs, without affecting the amplitude of sEPSCs. The glycine-induced increase in sEPSC frequency was blocked by 1 µM strychnine, a specific glycine receptor antagonist. The results suggest that glycine acts on presynaptic glycine receptors to increase the probability of glutamate release from excitatory nerve terminals. The glycine-induced increase in sEPSC frequency completely disappeared either in the presence of tetrodotoxin or Cd2+, voltage-gated Na+, or Ca2+channel blockers, suggesting that the activation of presynaptic glycine receptors might depolarize excitatory nerve terminals. The present results suggest that presynaptic glycine receptors can regulate the excitability of PAG neurons by enhancing glutamatergic transmission and therefore play an important role in the regulation of various physiological functions mediated by the PAG.

Summary Introduction Renal abnormality is not a rare finding in infants with primary VUR. The pathophysiology of the renal abnormality is considered to be congenital or acquired. Congenital hypo/dysplastic kidney is a common finding in infants with primary VUR, especially in boys. However, the long-term impact of unilateral hypo/dysplastic kidney has not been elucidated. The aim of the current study is to clarify the long-term impact of unilateral hypo/dysplastic kidney with primary vesicoureteral reflux diagnosed in infancy. Material and methods The medical records of patients with primary VUR detected in infancy with unilateral hypo/dysplastic kidney on initial nuclear renal scan ( Results Mean age at final visit was 12.4 years (range 5.9–22.2). Estimated GFR was evaluated in 26 patients at a mean age of 12.0 years (5.9–22.2). CKD stage was 1 in all. According to the guidelines of the Japanese Society of Hypertension, while none exceeded the standard level of systolic blood pressure (BP), two patients slightly exceeded the standard level of diastolic BP. In addition, no significant proteinuria was detected in all patients, although microalbuminemia was detected in 7.7% of patients. Discussion The prognosis of reflux nephropathy depends on the remnant renal tissue mass, that is, the number of normal nephrons. The normal congenital solitary kidney is reported to be hyperplastic with normal-sized glomeruli rather than hypertrophic ones with larger nephrons, and to have better long-term outcome regarding renal function. Accordingly, we speculated that patients with unilateral hypo/dysplastic kidney would have a similar number of nephrons to those without hypo/dysplastic kidney who have no or minimal scar as far as the contralateral kidney is well preserved. Long-term outcome of the current retrospective study was consistent with our speculation in terms of estimated GFR, proteinuria, or hypertension. Conclusions The present study demonstrated that significant clinical findings related to unilateral hypo/dysplastic kidney detected in infancy were rarely observed in the long term. Accordingly, unilateral hypo/dysplastic kidney seems to be a benign condition. To confirm this finding, further follow-up of these patients is necessary. Table . Clinical outcome in infants with primary VUR and unilateral hypo/dysplastic kidney. Estimated GFR at final evaluation (n = 26) 92.6–139.6 mL/min/1.73 m2, (CKD stage was 1 in all) Hypertension at final evaluation (n = 29) Systolic hypertension None Diastolic hypertension 2 Proteinuria at final evaluation Proteinuria on dipstick (n = 29) None Microalbuminuria (>30 mg/gCr) (n = 26) 2 (7.7%) (33.6 mg/gCr, 39.0 mg/gCr)

塩化ナトリウム添加によって得られる加熱処理WPI溶液のゲルについて,その形成過程における疎水性度およびSH量の変化について検討した.10%WPI溶液を70℃以上で加熱した場合,高分子化したホエータンパク質の可溶性凝集体が得られる.この過程においてホエータンパク質の疎水性度は上昇し,SH量は低下することがわかった.また,電気泳動分析から可溶性凝集体はSS結合を介していることがわかった.すなわち,加熱処理によりホエータンパク質問で疎水性相互作用およびSH/SS交換反応が生じ,可溶性凝集体が形成されるものと推察した.20℃において加熱処理WPI溶液に塩化ナトリウムを添加した場合,疎水性度の急激な上昇とわずかなSH量の低下をともないながらゲルが形成されることがわかった.このことからゲル形成には疎水性相互作用およびSH/SS交換反応の両方が関与しているが,初期段階で疎水性相互作用の方がより大きく関与していると考えた.

Recently, many cases of children presenting reversible splenial lesions during febrile illness (RESLEF) have been reported; however, their overall clinico-radiological features are unclear. To describe the clinico-radiological features, we retrospectively reviewed the etiology (pathogen), clinical course, laboratory data, magnetic resonance imaging and electroencephalography (EEG) findings, therapy, and prognosis of 23 episodes in 22 children (1 child recurred) who presented neurological symptoms, with RESLEF. The etiologies (pathogens) varied. Seizure occurred in 7 episodes, disturbance of consciousness (DC) in 13, and delirious behavior in 18. Serum sodium levels

Histaminergic neurons within the tuberomammillary nucleus (TMN) play an important role in sleep-wakefulness regulation. Here, we report the muscarinic modulation of GABAergic spontaneous miniature inhibitory postsynaptic currents (mIPSCs) in mechanically dissociated rat histaminergic neurons using a conventional whole-cell patch clamp technique. Muscarine, a nonselective muscarinic acetylcholine (mACh) receptor agonist, reversibly decreased mIPSC frequency without affecting the current amplitude, indicating that muscarine acts presynaptically to decrease the probability of spontaneous GABA release. The muscarine action on GABAergic mIPSC frequency was completely blocked by atropine, a nonselective mACh receptor antagonist, and tropicamide, an M(4) receptor antagonist. The muscarine-induced decrease in mIPSC frequency was completely occluded in the presence of Cd(2+), a general voltage-dependent Ca(2+) channel blocker, or in a Ca(2+)-free external solution. However, pharmacological agents affecting adenylyl cyclase or G-protein coupled inwardly rectifying K(+) channel activity did not prevent the inhibitory action of muscarine on GABAergic mIPSCs. These results suggest that muscarine acts on M(4) receptors on GABAergic nerve terminals projecting to histaminergic neurons to inhibit spontaneous GABA release via the inhibition of Ca(2+) influx from the extracellular space. Muscarine also inhibited action potential-dependent GABA release by activating presynaptic M(4) receptors in more physiological conditions. The M(4) receptor-mediated modulation of GABAergic transmission onto TMN neurons may contribute to the regulation of sleep-wakefulness.

INTRODUCTION AND OBJECTIVES: MAMLD1 (Mastermindlike domain containing 1) has been shown to be a causative gene for hypospadias. We have shown that transient knockdown of Mamld1 results in reduced testosterone production in mouse Leydig tumor cells (MLTC-1). However the mechanism underlying the regulation of the genes encoding steroidogenic enzymes remains unclear. We reported the subsequent clinical features of MAMLD1 mutation positive patients and further studied Mamld1 knockdown assay to elucidate this mechanism. METHODS: Three hypospadias patients who have MAMLD1 mutation were included in this study. We measured pituitary-gonadal serum hormone values and underwent ultrasonography. We performed transient knockdown assay with siRNAs for Mamld1 using MLTC-1, and measured amounts of steroid hormones, mRNA expressions of steroidogenic enzymes and cell proliferation rate. RESULTS: We identified nonsense mutations (E124X and R653X) which undergo nonsense mediated mRNA decay in these 3 patients with penoscrotal hypospadias. The patients who have E124X mutations are half-siblings. In clinical findings of 3 patients who have MAMLD1 mutation, whereas hormone values remained within the normal range at previous report, testosterone was low response to hCG stimulation and LH/FSH were elevated at the age of 7 and 9. Moreover testicular microlithiasis was detected by ultrasonography in 2 cases. By Mamld1 knockdown in MLTC-1, 17-hydroxypregnenolone, 17-hydroxyprogesterone and downstream metabolite products were reduced and the expressions of Cyp17a1 mRNA were significantly decreased. Although effect of Mamld1 knockdown by siRNA continued until 72hrs, number of viable cells did not change compared to control. CONCLUSIONS: We concluded that knockdown of Mamld1 reduce production of steroid hormones via regulation of steroidogenic enzymes gene expressions, particularly Cyp17a1. MAMLD1/Mamld1 is not only involved in testosterone production in fetal testis but also may be associated with adult testicular function.

WPI溶液を予め加熱処理し,室温で塩化ナトリウムを添加することによりゲル化する現象について検討し,以下のことが明らかとなった.(1) 80℃ 30分加熱処理した10% (w/w) WPI溶液に塩化ナトリウムを0.3-1.2% (w/w)添加すると,20℃において溶液は粘度の上昇をともないながら徐々にゲル化した.塩化ナトリウムの濃度が高くなるにしたがい粘度上昇は速く進み,ゲル化時間は短くなる傾向を示した.さらに,WPI溶液の加熱処理温度,塩化ナトリウムの濃度が高くなるにしたがい,ゲル強度は高くなることがわかった.(2) 電子顕微鏡観察の結果,WPI溶液を加熱処理することによりやや太く短い線状の可溶性凝集体が形成されることを確認した.さらに塩化ナトリウムの添加により,可溶性凝集体同士の会合が始まり,高分子化してゲルに至ることがわかった.塩化ナトリウム添加により得られたゲルは,GDL添加による酸性ゲルとよく似た網目状構造を呈していることがわかった.また,ゲルの網目状構造を形成している凝集体の太さは,塩化ナトリウム添加により得られたゲルの方がGDL添加による酸性ゲルよりもややランダムな構造をとることがわかった.

We present 4 cases of ossicular anomaly combined with anomalous tympanic membrane and/or stenosis of the external auditory canal. In 3 cases, a thick bony bridge connected the head of the malleus with the inferior posterior wall of the tympanic bone, and, in the other, a V-shaped ossicle surrounded by thick fibrous tissue was present inferior to the neck of the malleus. In 2 of the former 3 cases, a cartilageous bone was observed branching from the bony bridge anteriorly, the tip of which was clearly identified as the umbo of the malleus. Thus, the bony bridge was recognized as an anomaly which had developed on the anterior process of the malleus process of Folius in 3 cases, while in the other the malleus handle was completely absent. The area of the epithelial layer of the tympanic membrane was restricted due to partial protrusion of the tympanic bone in all cases. We speculate that the bridge anomaly developed at about the 19th week of gestation, when an intramembranous spicule (os goniale) directed towards the anteromedial surface of the immature cartilagenous malleus handle failed to fuse with it, and instead developed further to extend to the infero-posterior part of the tympanic bone. The V-shaped ossicle is thought to be an anomalous anterior process but not handle of the malleus because it is hard and does not contain any cartilaginous part characteristic to some part of the malleus handle.

The periaqueductal gray (PAG) is a major component of the descending pain inhibitory pathway, which is related to central analgesia. In the present study, we have investigated the possible roles of presynaptic nicotinic acetylcholine receptors in GABAergic transmission onto PAG neurons. In acutely isolated rat PAG neurons, GABAergic miniature inhibitory postsynaptic currents (mIPSCs) were recorded by use of a whole-cell patch clamp technique. Acetylcholine (30 microM) transiently increased both the frequency and amplitude of GABAergic mIPSCs. However, acetylcholine did not affect the GABA-induced membrane currents. This facilitatory action of acetylcholine disappeared in the presence of mecamylamine, a nonselective nicotinic receptor antagonist, and mimicked by nicotine, a nicotinic receptor agonist. The nicotine-induced increase in mIPSC frequency was completely blocked by dihydro-beta-erythroidine, a selective beta2-containing nicotinic receptor antagonist, but not methyllycaconitine or alpha-bungarotoxin, selective alpha7 nicotinic receptor antagonists. The results suggest that acetylcholine or nicotine acts presynaptic beta2-containing nicotinic receptors, presumably alpha4beta2 nicotinic receptors, to enhance spontaneous GABA release onto PAG neurons. The nicotine-induced increase in mIPSC frequency was completely occluded in the presence of Cd2+, a general voltage-dependent Ca2+ channels blocker, and in the absence of extracellular Ca2+ or Na+. The results suggest that presynaptic nicotinic receptors are less permeable to Ca2+, and that the activation of these receptors depolarizes GABAergic nerve terminals. In conclusion, presynaptic nicotinic receptors would temporally regulate the excitability of PAG neurons being not overexcited and eventually contribute to the cholinergic modulation of output from the PAG.

Japan A 56-yr-old Japanese male with a history of diabetic nephropathy underwent a HLA 5/6 mismatch and ABO-compatible living-related kidney transplantation (donor: his 49-yr-old wife). A pre-transplant standard NIH complement-dependent cytotoxicity cross-match (Xm) test, a flow-cytometric T-cell Xm, and a FlowPRA test were totally negative. Inductionimmunosuppressive protocol consisted of tacrolimus, mycophenolate mofetil, methylprednisolone, and basiliximab (BAS). The patient's post-operative course was almost uneventful, and the graft was functioning well (sCr 1.1 mg/dL). He developed general fatigue, and his sCr was elevated to 2.2 mg/dL 792 d after transplant. A graft biopsy showed acute T-cell mediated rejection Banff grade IB (i3, t3, g0, v0, ptc0, C4d staining negative). The conventional anti-rejection therapy could not improve his graft function; therefore, we added BAS to eliminate activated graft-infiltrating T-cells. He responded to the rescue therapy, and the improvement in graft function was confirmed by a subsequent graft biopsy. He enjoyed his health without any opportunistic infections.

Recent progress in genotyping technology and the development of public databases has enabled large-scale genome-wide association tests with diseases. We performed a two-stage genome-wide association study (GWAS) of bipolar disorder (BD) in Japanese cohorts. First we used Affymetrix 100K GeneChip arrays in the analysis of 107 cases with bipolar I disorder and 107 controls, and selected markers that were nominally significant (P < 0.01) in at least one of the three models (1,577 markers in total). In the follow-up stage, we analyzed these markers using an Illumina platform (1,526 markers; 51 markers were not designable for the platform) and an independent sample set, which consisted of 395 cases (bipolar I + II) and 409 controls. We also assessed the population stratification of current samples using principal components analysis. After the two-stage analysis, 89 markers remained nominally significant (allelic P < 0.05) with the same allele being consistently over-represented in both the first and the follow-up stages. However, none of these were significant after correction for multiple-testing by false discovery rates. Sample stratification was virtually negligible. Collectively, this is the first GWAS of BD in the Japanese population. But given the small sample size and the limited genomic coverage, these results should be taken as preliminary.

Although glycine receptors are found in most areas of the brain, including the hippocampus, their functional significance remains largely unknown. In the present study, we have investigated the role of presynaptic glycine receptors on excitatory nerve terminals in spontaneous glutamatergic transmission. Spontaneous EPSCs (sEPSCs) were recorded in mechanically dissociated rat dentate hilar neurons attached with native presynaptic nerve terminals using a conventional whole-cell patch recording technique under voltage-clamp conditions. Exogenously applied glycine or taurine significantly increased the frequency of sEPSCs in a concentration-dependent manner. This facilitatory effect of glycine was blocked by 1 microM strychnine, a specific glycine receptor antagonist, but was not affected by 30 microM picrotoxin. In addition, Zn(2+) (10 microM) potentiated the glycine action on sEPSC frequency. Pharmacological data suggested that the activation of presynaptic glycine receptors directly depolarizes glutamatergic terminals resulting in the facilitation of spontaneous glutamate release. Bumetanide (10 microM), a specific Na-K-2C co-transporter blocker, gradually attenuated the glycine-induced sEPSC facilitation, suggesting that the depolarizing action of presynaptic glycine receptors was due to a higher intraterminal Cl(-) concentration. The present results suggest that presynaptic glycine receptors on excitatory nerve terminals might play an important role in the excitability of the dentate gyrus-hilus-CA3 network in physiological and/or pathological conditions.

We report a 49-year-old man with a paraganglioma involving the nasopharynx, which according to the literature is the fourth case in Japan and the twentieth in the world. The initial histopathologic findings of the biopsy specimen from the nasopharyngeal tumor suspected a carcinoid tumor. However, the final histopathologic diagnosis from the surgically resected specimen was paraganglioma, determined by electron microscopy and using several stains, such as Grimelius and Neuron-specific enolase (NSE) stains. We wish to add this case of "primary" nasopharyngeal paraganglioma to the literature, and to report the pathologic findings and clinical course of this unusual case.

We investigated factors affecting the breakthrough urinary tract infection rate during prophylactic antibiotic treatment in children with primary vesicoureteral reflux.Medical charts were retrospectively reviewed in children with primary vesicoureteral reflux diagnosed at age 12 months or less who received prophylactic antibiotics and underwent (99m)Tc-dimercapto-succinic acid scan. Parameters assessed for their relation to breakthrough urinary tract infection were gender, presenting symptoms, age at presentation, prophylactic antibiotic type, reflux grade at presentation and scan findings.Enrolled in the study were 52 boys and 6 girls with a mean age at presentation of 3.7 months and a mean followup of 42.5 months. Urinary tract infection was a presenting symptom in 46 children. Low reflux grade (1-3) was identified in 18 children and 40 had high grade reflux (4-5). Abnormal (99m)Tc-dimercapto-succinic acid scan was documented in 36 children (62%). During followup breakthrough vesicoureteral reflux developed in 12 children, including 11 of 36 (31%) with an abnormal scan but only 1 of 22 (5%) with a normal scan (p = 0.021). The breakthrough urinary tract infection-free rate during followup was significantly lower in children with an abnormal scan (p = 0.033). Other factors were not significantly associated with the breakthrough urinary tract infection rate.Abnormal (99m)Tc-dimercapto-succinic acid scan may be a factor predicting breakthrough urinary tract infection in children with primary vesicoureteral reflux. Prophylactic antibiotics may have a limited treatment role in children with an abnormal scan.

Accelerated repopulation is a reason for loco-regional failure after radiotherapy for head and neck carcinoma. Accelerated fractionation(AF) is a radiotherapy regimen reducing the total treatment time, with the aim of counteracting tumor cell repopulation. AF administers the same or a similar total dose as conventional treatment in a reduced overall time by giving conventionally-sized or smaller fractions more than once daily. Several different clinical trials on AF have proved to be of benefit in loco-regional control, although no benefit in survival was generally detected. The metaanalysis of altered fractionated radiotherapy in head and neck cancer has showed a benefit with AF with conventional fractionation(CF). However, the magnitude of the survival benefit is lower with AF than with hyperfractionation (HF). In particular, AF using reduced total doses or a split course does not improve treatment benefits. AF that employs continuous RT schedules, without compromising the total dose, improves local control. More data on this AF regimen are needed. Acute morbidity is significantly more frequent with AF. Whether late toxicity is also worse with AF is unclear. Some trials suggest no increase in late toxicity, while others suggest the opposite. The effect of AF seems to be greater for the primary tumor than for the metastatic lymph-nodes. Also, the reduction of the treatment time is more beneficial in well- to moderately-differentiated tumors.

The spindle pole body (SPB) of Schizosaccharomyces pombe is required for assembly of the forespore membrane (FSM) during meiosis. Before de novo biogenesis of the FSM, the meiotic SPB forms outer plaques, an event referred to as SPB modification. A constitutive SPB component, Spo15, plays an indispensable role in SPB modification and sporulation. Here, we analyzed two sporulation-specific genes, spo13+and spo2+, which are not required for progression of meiotic nuclear divisions, but are essential for sporulation. Spo13 is a 16-kDa coiled-coil protein, and Spo2 is a 15-kDa nonconserved protein. Both Spo13 and Spo2 specifically associated with the meiotic SPB. The respective deletion mutants are viable, but defective in SPB modification and in the onset of FSM formation. Spo13 and Spo2 localized on the cytoplasmic side of the SPB in close contact with the nascent FSM. Localization of Spo13 to the SPB was dependent on Spo15 and Spo2; that of Spo2 depended only on Spo15, suggesting that their recruitment to the SPB is strictly controlled. Spo2 physically associated with both Spo15 and Spo13, but Spo13 and Spo15 did not interact directly. Taken together, these observations indicate that Spo2 is recruited to the SPB during meiosis and then assists in the localization of Spo13 to the outer surface of the SPB.

Profound activity-dependent synaptic facilitation at hippocampal mossy fiber synapses is a unique and functionally important property. Although presynaptic ionotropic receptors, such as kainate receptors, contribute partially to the facilitation in the hippocampus, the precise mechanisms of presynaptic regulation by endogenous neurotransmitters remain unclear. In this study, we report that axonal GABAAreceptors on mossy fibers are involved in the activity-dependent facilitation during development. In immature mouse hippocampal slices, short-train stimulation (five pulses at 25 Hz) caused frequency-dependent facilitation of not only postsynaptic responses but also presynaptic fiber volleys that represent presynaptic activities. This fiber volley facilitation was inhibited by selective GABAAreceptor antagonists, or by enkephalin that selectively suppresses excitability of interneurons. Furthermore, we directly demonstrated that this facilitation resulted from depolarization of mossy fibers in imaging experiments using a voltage-sensitive dye. This increased mossy fiber excitability caused by depolarizing action of GABA gradually decreased with development and eventually disappeared at around postnatal day 30. These results suggested that GABA released from interneurons acted on axonal GABAAreceptors on mossy fibers and contributed at least partially to the activity- and age-dependent facilitation in the hippocampus.

Several clinical reports have indicated that autistic patients often show disturbance of the circadian rhythm, which may be related to dysfunction of the serotonergic system in the brain. Using rats exposed prenatally to valproic acid (VPA) as an animal model of autism, we examined locomotor activity and feeding under a reversed 12-h light/dark cycle, and found disturbance of the circadian rhythm characterized by frequent arousal during the light/sleep phase. In addition, measurement of brain serotonin (5-HT) level using in vivo microdialysis showed that the brain 5-HT level in VPA-exposed rats was significantly higher than that in control rats. These results suggest that a higher brain 5-HT level might be responsible for the irregular sleep/awake rhythm in autism.

Dermatomyositis is an uncommon in flammatory myopathy with characteristic cutaneous manifestations which is frequently linked to several cancers. A 42-year-old man presented with left flank pain with typical symptoms of dermatomyositis. Computed tomography showed a solid mass in the pelvis of the left kidney and lymphadenopathy in the retroperitoneum. Since the general condition of the patient rapidly deteriorated because of marked muscle weakness of the respiratory muscles, the patient initially underwent medical treatment with prednisolone. After the improvement of respiratory function, the patient underwent radical nephroureterectomy and retroperitoneal lymphadenectomy. The pathological specimen showed moderately differentiated adenocarcinoma, stage pT3N2. After surgery, the patient showed a marked improvement of clinical symptoms related to dermatomyositis. Twenty-two months later, multiple organ metastases occurred and the patient died of cancer. We should point out the significance of surgical and medical treatment for these patients.

The demand for mobile bathing service (MBS) is increasing in the Japanese society. Therefore, we assessed the risk of MBS-associated infection in MBS clients and their caregivers by examining the bacterial colonization of MBS equipment and utensils. Bacterial isolates collected by the stamp agar culture method were examined by disk diffusion assay for their susceptibility to the following drugs: imipenem, ciprofloxacin, amikacin, azutreonam, ceftazidim, meropenem, piperacillin, tobramycin, ofloxacin and cefoperazone. Furthermore, these isolates were subtyped bySpeI-pulsed field gel electrophoresis (SpeI-PFGE). Fifty-fourP. aeruginosa isolates were recovered from different sampling sites, and of these, 26 (47.3%) were isolated from pillows. Eighteen isolates (33.3%) were imipenem (IPM) resistant. The minimum inhibitory concentrations (MICs) of 17 isolates were between 16 and 32 μg/ml, and the MIC of one isolate was greater than 32 μg/ml. TheSpeI-PFGE typing of IPM-resistant isolates revealed that 13 of the 18 isolates were closely related (F=1.0–0.87). Our findings suggest that MBS equipment and utensils, particularly pillows, are the primary sources of bacterial contamination and transmission and that there is a risk of MBS-mediated infection among MBS clients and their caregivers.

Conduct disorder (CD) is a serious psychiatric disorder of children and adolescents. A common association is the presence of attention-deficit hyperactivity disorder. The present paper reviews empirical findings on CD. The correlative factors were analyzed according to their bio-psycho-social aspects. To be effective, early detection and intervention on CD must be multimodal. We should address treatments on multiple foci, and continue them over an extensive period of time.

Upon activation of cell surface receptors coupled to the Gq subclass of G proteins, phospholipase C (PLC) beta hydrolyses membrane phospholipid to yield a pair of second messengers, inositol 1,4,5-trisphosphate (IP3) and 1,2-diacylglycerol. PLCbeta4 has been characterized as the isoform enriched in cerebellar Purkinje cells (PCs) and the retina and involved in motor and visual functions. Here we examined cellular and subcellular distributions of PLCbeta4 in adult mouse brains. Immunohistochemistry showed that high levels of PLCbeta4 were detected in the somatodendritic domain of neuronal populations expressing the metabotropic glutamate receptor (mGluR) type 1alpha, including olfactory periglomerular cells, neurons in the bed nucleus anterior commissure, thalamus, substantia nigra, inferior olive, and unipolar brush cells and PCs in the cerebellum. Low to moderate levels were detected in many other mGluR1alpha-positive neurons and in a few mGluR1alpha-negative neurons. In PCs, immunogold electron microscopy localized PLCbeta4 to the perisynapse, at which mGluR1alpha is concentrated, and to the smooth endoplasmic reticulum in dendrites and spines, an intracellular Ca2+ store gated by IP3 receptors. In the cerebellum, immunoblot demonstrated its concentrated distribution in the post-synaptic density and microsomal fractions, where mGluR1alpha and type 1 IP3 receptor were also greatly enriched. Furthermore, PLCbeta4 formed coimmunoprecipitable complexes with mGluR1alpha, type 1 IP3 receptor and Homer 1. These results suggest that PLCbeta4 is preferentially localized in the perisynapse and smooth endoplasmic reticulum as a component of the physically linked phosphoinositide signaling complex. This close molecular relationship might provide PLCbeta4 with a high-fidelity effector function to mediate various neuronal responses under physiological and pathophysiological conditions.

We investigated hippocampal substructure in the rat, cat, dog, and human by means of magnetic resonance imaging to elucidate phylogenetic differences in longitudinal organization. Multidirectional high-resolution images obtained with a 3 T scanner revealed that the dorsal part of the hippocampus was well developed in the rat, cat, and dog brain, and was homologous to the hippocampal tail, a poorly-developed posterior part, in the human. We conclude that the dorsal hippocampus of laboratory animals corresponds to the hippocampal tail in the human brain, which is considered to be hypoplastic and of less importance clinically than more anterior regions. These data may help in understanding phylogenetic, and in correlating results from animal experiments with clinical findings on the functions and pathologies of the human hippocampus.

Basic studies were carried out to apply frozen allogeneic nerve grafts in dogs after wide-ranging defects of the brachial plexus due to surgical resection of tumor. In this study, morphological variations in branching patterns of the brachial plexus were examined in ten beagle dogs, to evaluate whether the brachial plexus might represent a useful source of allogeneic nerve grafts. Spatial relationships between the axillary lymph node, which had the possibility of carcinomatous metastasis, and the musculocutaneous (MC) nerve, which was important for the function of the forelimbs, were also investigated. In all ten cases examined, the brachial plexus received ventral roots from the fifth cervical nerve to the first thoracic nerve. No significant variation in the branching pattern was found in any nerve except the phrenic, MC and dorsal thoracic nerves. Four communicating branches were observed and had some morphological variations which might be negligible for nerve grafting. Considering previous physiological and anatomical reports, the most important nerve to be reunited in graft operations for functional recovery is the radial nerve. The MC nerve and median or ulnar nerve should also be considered as possibilities for reuniting. Distances between the axillary lymph nodes and the MC nerve ranged from 11.2 mm to 21 mm (mean +/- SD: 16.1 +/- 2.3 mm). In conclusion, it was suggested that morphological variations in the brachial plexus were technically acceptable to apply allogeneic nerve grafts at least in beagle dogs.

We studied the allelic distributions of 48-base pair repeat polymorphism of the dopamine D4 receptor (DRD4) gene in two groups of schizophrenic patients, one with any schizophrenic member in the family (L(+) group) and the other without schizophrenic family histories (L(-) group), and healthy controls. The frequency of the sum of the alleles of 4, 5, and 6 repeats in the L(+) group was significantly higher than the controls. The frequency of the allele of 4 repeats in the L(-) group was significantly higher than the controls. These results show positive allelic associations between DRD4 polymorphism and each of the two groups of schizophrenic patients.

Mepanipyrim inhibited retrograde Golgi-to-ER trafficking induced by brefeldin A (BFA), nordihydroguaiaretic acid, clofibrate, and arachidonyltrifluoromethyl ketone in NRK and other types of cells, but did not inhibit anterograde trafficking of Golgi-resident proteins translocated to ER by BFA and newly synthesized VSV-G. However, mepanipyrim did not block the TGN38 dispersion induced by any of these compounds. Mepanipyrim acted on the Golgi, and swollen vesicular Golgi structures were formed and similar structures accumulated during rebuilding of the Golgi after BFA removal. These actions of mepanipyrim were readily reversed after its removal. Mepanipyrim did not stabilize microtubules, but prevented nocodazole-induced fragmentation and dispersion of the Golgi. These results suggest that the mepanipyrim-sensitive molecules participated in stabilizing the Golgi and its anchoring in the perinuclear region, and equally importantly, that the novel action of mepanipyrim may be used as a pharmacological tool for investigating membrane transport, Golgi membrane dynamics, and differentiation of the Golgi from TGN.

The Schizosaccharomyces pombe spo14-B221 mutant was originally isolated as a sporulation-deficient mutant. However, thespo14+gene is essential for cell viability and growth. spo14+is identical to the previously characterizedstl1+gene encoding a putative homologue of Saccharomyces cerevisiae Sec12, which is essential for protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus. In the spo14 mutant cells, ER-like membranes were accumulated beneath the plasma membrane and the ER/Golgi shuttling protein Rer1 remained in the ER. Sec12 is a guanine nucleotide exchange factor for the Sar1 GTPase. Overproduction ofpsr1+coding for an S. pombe Sar1 homologue suppressed both the sporulation defect ofspo14-B221 and cold-sensitive growth of newly isolatedspo14-6 and spo14-7 mutants. These results indicate that Spo14 is involved in early steps of the protein secretory pathway. The spo14-B221 allele carries a single nucleotide change in the branch point consensus of the fifth intron, which reduces the abundance of the spo14 mRNA. During meiosis II, the forespore membrane was initiated near spindle pole bodies; however, subsequent extension of the membrane was arrested before its closure into a sac. We conclude that Spo14 is responsible for the assembly of the forespore membrane by supplying membrane vesicles.