Kidney enlargement and multiple liver cyst formation implicate mutations in PKD1/2 in adult sporadic polycystic kidney disease.

Distinguishing autosomal-dominant polycystic kidney disease (ADPKD) from other inherited renal cystic diseases in patients with adult polycystic kidney disease and no family history is critical for correct treatment and appropriate genetic counseling. However, for patients with no family history, there are no definitive imaging findings that provide an unequivocal ADPKD diagnosis. We analyzed 53 adult polycystic kidney disease patients with no family history. Comprehensive genetic testing was performed using capture-based next-generation sequencing for 69 genes currently known to cause hereditary renal cystic diseases including ADPKD. Through our analysis, 32 patients had PKD1 or PKD2 mutations. Additionally, 3 patients with disease-causing mutations in NPHP4, PKHD1, and OFD1 were diagnosed with an inherited renal cystic disease other than ADPKD. In patients with PKD1 or PKD2 mutations, the prevalence of polycystic liver disease, defined as more than 20 liver cysts, was significantly higher (71.9% vs 33.3%, P = .006), total kidney volume was significantly increased (median, 1580.7 mL vs 791.0 mL, P = .027) and mean arterial pressure was significantly higher (median, 98 mm Hg vs 91 mm Hg, P = .012). The genetic screening approach and clinical features described here are potentially beneficial for optimal management of adult sporadic polycystic kidney disease patients.

Mouth breathing, another risk factor for asthma: the Nagahama Study.

BACKGROUND: Allergic rhinitis, a known risk factor for asthma onset, often accompanies mouth breathing. Mouth breathing may bypass the protective function of the nose and is anecdotally considered to increase asthma morbidity. However, there is no epidemiological evidence that mouth breathing is independently associated with asthma morbidity and sensitization to allergens. In this study, we aimed to clarify the association between mouth breathing and asthma morbidity and allergic/eosinophilic inflammation, while considering the effect of allergic rhinitis. METHODS: This community-based cohort study, the Nagahama Study, contained a self-reporting questionnaire on mouth breathing and medical history, blood tests, and pulmonary function testing. We enrolled 9804 general citizens of Nagahama City in the Shiga Prefecture, Japan. RESULTS: Mouth breathing was reported by 17% of the population and was independently associated with asthma morbidity. The odds ratio for asthma morbidity was 1.85 (95% CI, 1.27-2.62) and 2.20 (95% CI, 1.72-2.80) in subjects with mouth breathing alone and allergic rhinitis alone, which additively increased to 4.09 (95% CI, 3.01-5.52) when mouth breathing and allergic rhinitis coexisted. Mouth breathing in nonasthmatics was a risk for house dust mite sensitization, higher blood eosinophil counts, and lower pulmonary function after adjusting for allergic rhinitis. CONCLUSION: Mouth breathing may increase asthma morbidity, potentially through increased sensitization to inhaled allergens, which highlights the risk of mouth-bypass breathing in the 'one airway, one disease' concept. The risk of mouth breathing should be well recognized in subjects with allergic rhinitis and in the general population.

Association of genetic variation in FTO with risk of obesity and type 2 diabetes with data from 96,551 East and South Asians.

AIMS/HYPOTHESIS: FTO harbours the strongest known obesity-susceptibility locus in Europeans. While there is growing evidence for a role for FTO in obesity risk in Asians, its association with type 2 diabetes, independently of BMI, remains inconsistent. To test whether there is an association of the FTO locus with obesity and type 2 diabetes, we conducted a meta-analysis of 32 populations including 96,551 East and South Asians. METHODS: All studies published on the association between FTO-rs9939609 (or proxy [r (2) > 0.98]) and BMI, obesity or type 2 diabetes in East or South Asians were invited. Each study group analysed their data according to a standardised analysis plan. Association with type 2 diabetes was also adjusted for BMI. Random-effects meta-analyses were performed to pool all effect sizes. RESULTS: The FTO-rs9939609 minor allele increased risk of obesity by 1.25-fold/allele (p = 9.0 × 10(-19)), overweight by 1.13-fold/allele (p = 1.0 × 10(-11)) and type 2 diabetes by 1.15-fold/allele (p = 5.5 × 10(-8)). The association with type 2 diabetes was attenuated after adjustment for BMI (OR 1.10-fold/allele, p = 6.6 × 10(-5)). The FTO-rs9939609 minor allele increased BMI by 0.26 kg/m(2) per allele (p = 2.8 × 10(-17)), WHR by 0.003/allele (p = 1.2 × 10(-6)), and body fat percentage by 0.31%/allele (p = 0.0005). Associations were similar using dominant models. While the minor allele is less common in East Asians (12-20%) than South Asians (30-33%), the effect of FTO variation on obesity-related traits and type 2 diabetes was similar in the two populations. CONCLUSIONS/INTERPRETATION: FTO is associated with increased risk of obesity and type 2 diabetes, with effect sizes similar in East and South Asians and similar to those observed in Europeans. Furthermore, FTO is also associated with type 2 diabetes independently of BMI.

Tooth loss and atherosclerosis: the Nagahama Study.

Several epidemiologic studies have suggested that oral disease is a risk factor for cardiovascular disease (CVD). However, whether a clinically significant association exists between the 2 disorders remains controversial. Here, we investigated the association between tooth loss, as an indicator of oral disease, and arterial stiffness, as a marker of atherosclerosis, in Japanese adults. Cross-sectional data were collected for 8,124 persons aged 30 to 75 y with no history of tooth loss for noninflammatory reasons, such as orthodontic treatment, malposition, and trauma. Participants received a comprehensive dental examination and extensive in-person measurements of CVD risk factors, and arterial stiffness was evaluated using the cardio-ankle vascular index (CAVI). We examined the association between CAVI and tooth loss using general linear models with adjustment for age, sex, body mass index, smoking status, hemoglobin A1c, and a history of insulin or hypoglycemic medication depending on the model. In addition, we performed an analysis that included interaction terms of the centered variables tooth loss, sex, and age. The results of the multiple regression analysis that included the interaction terms detected that the relationship between CAVI and tooth loss was dependent on sex, with only men showing a positive correlation (ß for interaction = 0.04; 95% confidence interval, 0.02-0.06). The findings from this study suggest that a linear relationship exists between tooth loss and degree of arterial stiffness and that the association differed depending on sex.

Marked suppression of cortical auditory evoked response shortly before the onset of REM sleep.

In 10 of 12 subjects examined, the amplitude of N300, a component of the cortical auditory evoked potential, was evidently smaller in rapid eye movement (REM) sleep than in non-REM sleep. The start of the reduction associated with the onset of the first episode of REM sleep was examined in these 10 subjects. In five of these, a marked reduction of N300 amplitude occurred 0.5-2.5 min before the appearance of muscle atonia of REM sleep. In two subjects, a similarly marked reduction of the N300 amplitude occurred 0.5-1.0 min before the disappearance of sleep spindles or K-complexes. This suggests that a suppression of the synchronizing mechanism in the cerebrum sometimes occurs briefly prior to the occurrence of other physiological phenomena associated with REM sleep.

Effects of hypoxia on stimulus-release coupling mechanisms in cultured bovine adrenal chromaffin cells.

To clarify the effects of hypoxia on stimulus-release coupling, we have examined the effects of hypoxia on nicotine-induced catecholamine (noradrenaline and adrenaline) release from, and 22Na+ influx, 45Ca2+ influx and cytosolic free Ca2+ concentration ([Ca2+]i) in, cultured bovine adrenal chromaffin cells. Experiments were carried out in media pre-equilibrated with 21% O2/79% N2 (control) or with 0% O2/100% N2 (hypoxia). Cells were stimulated with either nicotine (activating nicotinic acetylcholine (ACh) receptors) or a high K+ concentration (55 mmol/l KCl; directly activating voltage-dependent Ca2+ channels). Hypoxia reduced both nicotine- and high K(+)-induced catecholamine releases from the cells, but the reduction of the former (to about 30% of the control value) was more pronounced than that of the latter (to about 40% of the control value). Nicotine-induced 22Na+ influx, which is considered to reflect the function of nicotinic ACh receptors, was inhibited by hypoxia. Both nicotine- and high K(+)-induced 45Ca2+ influx into the cells were reduced by hypoxia, but the reduction of the former was more pronounced than that of the latter. Nicotine- and high K(+)-induced increases in [Ca2+]i were reduced by hypoxia to about 30% and 40% of the control values, respectively. These results suggest that hypoxia reduces cation influxes (Na+ and Ca2+) through both the ligand-gated cation channels of the nicotinic ACh receptor and the voltage-dependent Ca2+ channels.

Structure of NG-061, a novel potentiator of nerve growth factor (NGF) isolated from Penicillium minioluteum F-4627.

The structure of NG-061, a new potentiator of nerve growth factor (NGF) isolated from Penicillium minioluteum F-4627, was determined by spectroscopic analysis and X-ray diffraction method to be phenylacetic acid 2-(2-methoxy-4-oxocyclohexa-2,4-dienylidene)-hydrazide.

CUL1, a component of E3 ubiquitin ligase, alters lymphocyte signal transduction with possible effect on rheumatoid arthritis.

Ubiquitination affects various immune processes and E3 ubiquitin ligases (E3) play an important role in determining substrate specificity. We identified 11 human E3 ligase genes of potential importance in pathogenesis of autoimmune diseases by search of public databases and screened them for candidacy of biological investigation with case-control linkage disequilibrium tests on multiple SNPs in the genes using rheumatoid arthritis (RA) as a model of autoimmune diseases. Significant association with RA was observed in an SNP in intron 3 of Cullin 1 (CUL1) that affected transcriptional efficiency of the promoter activity in lymphocytic cell lines. Quantitative expression analysis revealed that CUL1 mRNA was highly detected in lymphoid tissues including spleen and tonsil, and was specifically expressed in T and B lymphocytes in fractionated peripheral leukocytes. Histological evaluation of tonsils indicated that CUL1 protein expression was relatively specific for maturing germinal centers. Suppression of CUL1 expression had influence on the phenotype of T-cell line, that is, it inhibited IL-8 induction, which is known to play an important role in the migration of inflammatory cells into the affected area seen in RA. Our data suggest that the regulation of CUL1 expression in immunological tissues may affect the susceptibility of RA via altering lymphocyte signal transduction.

Single nucleotide polymorphisms in the gene encoding Krüppel-like factor 7 are associated with type 2 diabetes.

AIMS/HYPOTHESIS: Although genetic susceptibility plays an important role in the pathogenesis of type 2 diabetes, most of the genes that influence susceptibility to type 2 diabetes have yet to be identified. Krüppel-like transcription factors are known to play important roles in development and cell differentiation, and have recently been implicated in the pathogenesis of type 2 diabetes. The present study aimed to examine the associations of single nucleotide polymorphisms (SNPs) in genes encoding members of the Krüppel-like-factor (KLF) family with type 2 diabetes in a large cohort of Japanese subjects. METHODS: We genotyped 33 SNP loci found in 12 KLF genes in subjects with type 2 diabetes and in subjects from the general population using the PCR-Invader assay. We also examined the effects of the overexpression of KLF7 on adipogenesis in 3T3-L1 cells. RESULTS: We identified a significant association between an SNP in KLF7 and type 2 diabetes (A vs C: p=0.004 after Bonferroni's correction, odds ratio=1.59, 95% CI 1.27-2.00). The expression of Klf7 decreased in response to the differentiation of 3T3-L1 adipocytes, and the overexpression of KLF7 resulted in significant inhibition of adipogenesis in 3T3-L1 cells. CONCLUSIONS/INTERPRETATION: These results indicate that the gene encoding KLF7 is a novel candidate for conferring genetic susceptibility to type 2 diabetes.

Possible mechanisms of inhibitory action of sodium nitroprusside on histamine-induced contractile responses in isolated rabbit aorta compared with basilar artery and taenia coli.

Possible mechanisms of the inhibitory actions of sodium nitroprusside (NaNP) on histamine-induced contractile responses of isolated rabbit aorta, basilar artery and taenia coli were studied. NaNP (3 X 10(-5) M) reduced maximum contractile response to histamine in the aorta, whereas a concentration (10(-4) M) of NaNP slightly shifted the concentration-response curve for histamine towards its higher concentrations, but did not influence the maximum response in the basilar artery and taenia coli. The reduction in the maximum response of the aorta by NaNP was reversed by removal of the endothelium, or treatment with indomethacin (5 X 10(-6) M) or quinacrine (5 X 10(-6) M), but not by nordihydroguaiaretic acid (NDGA) (5 X 10(-5) M). Prostaglandin (PG) E1, PGE2 and PGI2 produced a relaxation in histamine-contracted basilar artery and taenia coli, whereas PGE1 and PGE2 produced a weak relaxation in histamine-contracted aorta. PGD2 relaxed taenia coli contracted by histamine, but had no influences in aorta and basilar artery. From these results, it is concluded that the inhibitory action of NaNP on histamine-induced contractile response in isolated rabbit aorta may be mediated at least partly by endothelium-derived arachidonic acid metabolite(s) via the cyclooxygenase pathway which must not involve PGI2. Furthermore, the difference in the inhibitory actions of NaNP between the tissues used in this study must be attributed to the absence of such mechanisms both in the basilar artery and taenia coli.

Comparison between magnifying endoscopy and histological, culture and urease test findings from the gastric mucosa of the corpus.

BACKGROUND AND STUDY AIMS: The incidence of Helicobacter pylori infection in Japan is high. Unlike the case in Western countries, H. pylori-induced gastritis in Japanese patients has a tendency to spread to the corpus. H. pylori-induced gastritis is characterized by a number of specific endoscopic findings. In a previous study, the endoscopic features and a magnified view of the gastric mucosa of the corpus of H. pylori-negative normal stomach were described. This report describes the specific histological features and magnified views of the H. pylori-negative stomach and compares them with those seen during H. pylori-induced gastritis. PATIENTS AND METHODS: The anterior wall or greater curvature of the middle body of the stomachs of 297 patients were observed by magnifying endoscopy (x 80). Forceps biopsy was performed at the following locations: i) the magnified site, for histological examination; ii) the antral mucosa, for culture/urease test, and histology; and iii) greater curvature of the upper body for culture/urease test. RESULTS: 72 patients were diagnosed as having H. pylori-negative normal stomach and 225 as having H. pylori-positive gastritis. The magnified views were classified into four types: i) collecting venules, with true capillaries forming a network, and gastric pits resembling pinholes (type Z-0; n = 80); ii) irregular true capillaries but no collecting venules observed (type Z-1; n = 36); iii) white gastric pits and sulci, with neither collecting venules nor true capillaries being seen (Z-2; n = 110); and iv) dilated pits with surrounding redness (Z-3; n = 71). All cases of H. pylori-negative normal stomach were type Z-0, whereas H. pylori-induced gastritis was present in all cases where the classification was Z-1, Z-2, or Z-3. Type Z-0 differed significantly from types Z-1, Z-2, and Z-3 with regard to the grade of inflammation, activity, and presence of H. pylori. CONCLUSIONS: Collecting venules and true capillaries forming a network with gastric pits in the center (type Z-0) were observed in the H. pylori-negative normal mucosa. The magnified views of H. pylori-related gastritis clearly differed from type Z-0.

Substrate for botulinum ADP-ribosyltransferase, Gb, has an amino acid sequence homologous to a putative rho gene product.

Gb (b for botulinum) is an Mr 21,000-22,000 guanine nucleotide binding protein which is specifically ADP-ribosylated by an ADP-ribosyltransferase produced by types C1 and D Clostridium botulinum (Morii, N., Sekine, A., Ohashi, Y., Nakao, K., Imura, H., Fujiwara, M., and Narumiya, S. (1988) J. Biol. Chem. 263, 12420-12426). In this study, we have identified the partial amino acid sequence of this protein. Purified Gb was digested with various proteases, and the proteolytic fragments were separated by high performance liquid chromatography. Isolated peptides were subjected to protein sequencing by an automated pulse-liquid phase protein sequenator. Eight partial amino acid sequences consisting of a total of 105 amino acid residues were obtained from 16 peptides. Computerized homology analysis revealed that they are homologous to the sequence deduced from cDNA of the rho genes. About 88% of the amino acid residues was identical between the sequenced Gb fragments and the corresponding regions of the Aplysia rho gene product. The result strongly suggests that Gb is a putative rho gene product.

Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase.

We reported previously that the ADP-ribosyltransferase in C1 and D botulinum toxins specifically catalyzes ADP-ribosylation of an Mr 22,000 guanine nucleotide-binding protein and that this substrate named Gb (b = botulinum) has an amino acid sequence homologous to that deduced from the rho gene (Narumiya, S., Sekine, A., and Fujiwara, M. (1988) J. Biol. Chem. 263, 17255-17257). In this study we have determined the amino acid sequence at its ADP-ribosylation site. Purified substrate was [32P]ADP-ribosylated by C1 botulinum toxin and digested with trypsin. The radioactive peptides were isolated by reversed-phase high performance liquid chromatography and digested further either with protease V8, with proteases V8 and thermolysin, or with proline endopeptidase and thermolysin. By this procedure three radioactive peptides were obtained, and their amino acid sequences were X-Tyr-Val-Ala-Asp-Ile-Glu, X-Tyr, and Val-Phe-Glu-X-Tyr in which no amino acid peak was found in X. During the sequencing the radioactivity quantitatively adhered to the sequencing filter and was not eluted with either of the identified amino acid residues. Analysis of the protein without the ADP-ribosylation yielded the corresponding sequence as Thr-Val-Phe-Glu-Asn-Tyr which corresponds to Thr37-Tyr42 in the amino acid sequence deduced from the Aplysia rho gene. These results strongly suggest that the asparagine residue is the ADP-ribosylation site in the rho gene product. This ADP-ribose protein bond was stable in 0.5 M hydroxylamine at pH 7.5 at 37 degrees C for at least 5 h. The ADP-ribosylation of this protein affected neither its GTPase- nor its [35S]guanosine 5'-O-thiotriphosphate-binding activity.

Axial eye-length measurement by wavelength-shift interferometry.

A simplified noncontact measuring technique for axial eye length was developed. According to this method, the wavelength shift of a single-mode laser-diode beam that is irradiated onto the eyeball causes a phase shift in the interference fringes of reflections from the retina and the cornea. Then the optical distance between the cornea and the retina is obtained from the phase-shift measurement. High-speed axial eye-length measurements can be performed by using a laser diode on a pulse-modulation drive and signals from the reference light path as an analog-to-digital-conversion trigger. Compared with the technique that uses partially coherent light, this technique is inferior in terms of measurement accuracy but superior in its wide, measurable range of 16-32 mm. The results of measurements of 21 adults showed that 2 standard deviations of measurement was 2 sigma = +/- 0.11 mm.

Benign bilateral independent periodic lateralized epileptiform discharges.

Bilateral independent periodic lateralized epileptiform discharges (BIPLEDs) usually appear transiently in patients with severe disturbances of consciousness and are indicative of a poor prognosis. Recurrent BIPLEDs have not previously been reported in the literature. We report a 64-year-old patient with bilateral hippocampal lesions (cerebral infarction) who exhibited persistent periodic lateralized epileptiform discharges (PLEDs) (chronic PLEDs) associated with recurrent BIPLEDs. Electroencephalography was recorded for more than 6 months. Left hemispheric PLEDs appeared first. Next, PLEDs shifted to the right hemisphere and BIPLEDs occasionally developed. Brain magnetic resonance imaging and single-photon emission computed tomography with technetium-99-hexamethyl-propyleneamine oxime was performed before and after the appearance of BIPLEDs. The patient had no remarkable clinical symptoms aside from mild memory impairment for this period of time. This is the first case of recurrent 'benign' BIPLEDs, that is, BIPLEDs with a positive prognosis.

ADP-ribosylation of the rho/rac gene products by botulinum ADP-ribosyltransferase: identity of the enzyme and effects on protein and cell functions.

1. Botulinum C1 toxin and C3 exoenzyme were purified from the culture filtrate of type C Clostridium botulinum strain 003-9, and specific antibodies were raised against each protein. Immunochemical analysis using these antibodies revealed the presence of minute amount of a C3-like molecule in C1 toxin preparation which tightly binds to the toxin component(s). This enzyme complex was separated from the major neurotoxin. Thus, the ADP-ribosyltransferases in C1 and D toxins and C3 exoenzyme appear to come from the same origin, and should be called together botulinum C3 enzyme. 2. Botulinum C3 enzyme ADP-ribosylates the rho and rac gene products, a family of small molecular weight GTP-binding proteins homologous to ras p21s. This ADP-ribosylation occurs at Asn41 of the rho products which is located in their putative effector domain, suggesting that it interferes interaction of these GTP binding proteins with their effector molecules. 3. When incubated with PC-12 cells, the enzyme inhibits cell growth and induces neurites and acetylcholine esterase. Several lines of evidence suggest that the ADP-ribosylation of the rho/rac proteins is responsible for these changes.

K-complex evoked in NREM sleep is accompanied by a slow negative potential related to cognitive process.

Evoked cortical responses to two kinds of auditory stimuli (rare and frequent) were analyzed to determine whether or not a K-complex evoked in stage 2 of NREM sleep is accompanied by some endogenous cognitive components of the event-related potential. All the 7 subjects examined in this sleep state failed to provide the correct behavioral response to auditory stimuli, but a K-complex was evoked more frequently by rare stimuli than by frequent stimuli. EEG segments in stage 2 were averaged separately according to the presence or absence of K-complexes emerging just after the stimulation. In cases where K-complexes did not emerge, a long-lasting negative potential of relatively low voltage appeared in the difference wave, which was obtained by subtracting the averaged EEG for frequent stimuli from that for rare stimuli. In cases where K-complexes emerged, a similar long-lasting negative potential of large amplitude appeared in the difference wave. These data may indicate that a K-complex evoked by an external stimulus is accompanied by a potential related to a cognitive process, which appears with greater amplitude in cases where a K-complex is evoked.

Cortical activity of REM sleep often occurs earlier than other physiological phenomena.

N300 appearing in response to sound stimulus was used as an index to determine the occurrence of cortical activity characterizing REM sleep. In 5/10 subjects, marked reduction of N300 amplitude occurred even in the period of 0.5-2.5 min immediately preceding the appearance of muscle atonia characterizing REM sleep. Neither muscle atonia nor rapid eye movements appeared prior to the marked reduction of N300 amplitude in any subject. This suggests that the cortical activity characterizing REM sleep sometimes occurs a few minutes (or less) earlier than other physiological phenomena.

Nonsteroidal anti-inflammatory drug-associated colitis with a histology of collagenous colitis.

Here we report a case of nonsteroidal anti-inflammatory drug (NSAID)-associated colitis with a histology of collagenous colitis in a 77-year-old woman. The patient had taken aspirin since 1993 after being diagnosed at another hospital, as having multiple cerebral infarctions. She began to suffer from intermittent diarrhea in April 1999. Serological examination showed hypoproteinemia, which indicated that she had protein-losing enteropathy. By July 1999, she had undergone colonoscopic examination four times. Biopsy specimens taken during the fourth colonoscopy revealed collagenous colitis. As the patient had been taking aspirin for 6 years, she was diagnosed as having NSAID-associated colitis with a histology of collagenous colitis. When she stopped taking aspirin, the diarrhea ceased. Three months later, the patient underwent a fifth colonoscopy. A histological examination of the biopsy specimen revealed that the collagen band had vanished. NSAID-associated colitis sometimes shows collagenous colitis histologically and is cured by withdrawing the drug. It is important to differentiate NSAID-associated colitis, even if it shows a histology of collagenous colitis, from collagenous colitis as the two diseases differ in etiology and therapy.

A negative component superimposed on event-related potentials during light drowsiness.

The present study examined the nature of the negative shift of event-related potential (ERP) recorded during the fully awake state, wakefulness with minor awareness deficit (light drowsiness) and stage 1 of NREM sleep. The cortical responses evoked by two types of auditory stimuli were recorded in nine subjects at the different levels of consciousness described above. A negative component with peak latency of 250-350 msec, N300, was identified in ERP during light drowsiness but not in the fully awake state. In stage 1a (stage 1 without vertex sharp waves), the amplitude of N300 was higher than that in light drowsiness, and it was higher in stage 1b than in stage 1a. The scalp distribution of N300 was predominantly on the vertex. It also confirmed that the vertex sharp wave evoked during stage 1 is maximal on the vertex and its peak latency is approximately 300 msec. Considering the similarity in scalp distribution and peak latency between N300 and vertex sharp wave, it is possible that these electroencephalogram phenomena are generated by an identical synchronizing mechanism in the brain. We assumed that N300 observed during light drowsiness may be an incomplete product of vertex sharp wave.