A trans-ethnic genetic study of rheumatoid arthritis identified FCGR2A as a candidate common risk factor in Japanese and European populations

Rheumatoid arthritis (RA) is a common systemic autoimmune disease and its onset and prognosis are controlled by genetic, immunological, and environmental factors. The HLA locus, particularly HLA-DRB1, is its strongest genetic risk determinant across ethnicities. Several other genes, including PTPN22 and PADI4, show modest association with RA. However, they cover only a part of its genetic components and their relative contribution is different between populations. To identify novel genetic determinants, we took a candidate gene approach in a trans-ethnic manner. After critical selection of 169 genes based on their immunological function, we performed SNP discovery of these genes by the resequencing of exons and surrounding areas using European and Japanese DNAs. We then generated a panel of 1,509 SNPs for case?Ccontrol association study in both populations. The DerSimonian?CLaird test for meta-analysis, using the combined results of the two populations, identified rs7551957 at the 5??-flanking region of the low-affinity Fc-gamma receptor IIa (FCGR2A) gene as the strongest candidate for the association (p?=?8.6?×?10^?5, odds ratio?=?1.58 with 95%CI 1.25?C1.99). Suggestive signals were also obtained for three SNPs in the dihydropyrimidine dehydrogenase (DPYD) gene (rs6685859; p?=?1.3?×?10^?4, rs7550959; p?=?1.5?×?10^?4 and rs7531138; p?=?1.7?×?10^?4) and an intronic SNP, rs2269310, of the erythrocytic spectrin beta (SPTB) gene (p?=?7.9?×?10^?4).     

Nutritional effects of supplementing liquid-formula diet with dietary fiber on elderly bed-ridden patients

In the past few decades, the number of bed-ridden elderly patients has been increasing. This group of patients is frequently fed with a liquid formula diet. The aim of this study was to evaluate the usefulness of a liquid formula diet containing dietary fiber (DF) for elderly bed-ridden patients. Eighteen elderly, bed-ridden patients were given L-3 Fiber, a DF-containing liquid formula diet (DF-LFD), for 4 weeks, while a number of parameters were monitored, including serum levels of total cholesterol, triglyceride, total protein, creatinine, uric acid, glucose, sodium, potassium, and calcium, urine protein/sugar, and defecation frequency. Total protein, albumin and total cholesterol significantly increased following the administration of the DF-LFD, associated with an average increase in body weight of 1.94 kg (5.0%). Defecation frequency significantly increased one week after DF-LFD administration was started, but this effect was transient. Although a few patients complained of nausea, vomiting or abdominal pain, no severe side effects were seen. In conclusion, DF-LFD supplementation appears to be beneficial for elderly bed-ridden patients, and can increase nutritional-related parameters, such as body weight, total protein, albumin and total cholesterol, without severe side effects.     

Thrombotic thrombocytopenic purpura in a child with systemic lupus erythematosus

We report a child with thrombotic thrombocytopenic purpura (TTP) secondary to systemic lupus erythematosus. The diagnosis was confirmed by low ADAMTS13 activity (<5%) along with the presence of a low titer inhibitor. Her clinical course was complicated by systemic lupus erythematosus, immunosuppressant therapy, and septic shock. She responded to plasma exchange and ADAMTS13 activity levels recovered. This case illustrates the heterogeneity of TTP and the difficulty of making a diagnosis of TTP. ADAMTS13 activity assay can be useful in the differential diagnosis of diseases with clinical features of thrombotic microangiopathy in pediatric patients. However, treatment needs to be decided carefully case-by-case.     

Clinical study of transcutaneous vaccination using a hydrogel patch for tetanus and diphtheria

Transcutaneous immunization (TCI) is a non-invasive and easy-to-use vaccination method. We demonstrated the efficacy and safety of a transcutaneous vaccine formulation using a hydrogel patch in animal experiments. In the present study, we performed a clinical study to apply our TCI formulation for vaccination against tetanus and diphtheria in human. The TCI device was a hydrogel patch (antigen-free) applied to the left brachial medial skin of 22 healthy volunteers for 48 h. Next, the hydrogel patch, containing 2mg tetanus toxoid (TT) and 2mg diphtheria toxoid (DT) as the TCI formulation, was applied to 27 healthy volunteers for 24h and some volunteers were vaccinated again by TCI formulation. For safety assessment, the patch application site was observed to assess local adverse events, and systemic adverse events were determined by a blood test. The antigen-free hydrogel patch and TCI formulation containing TT and DT did not induce local or systemic severe adverse events. For vaccine efficacy estimation, toxoid-specific serum antibody titers were determined by ELISA and the toxin-neutralizing activity of the induced antibody was evaluated in a passive-challenge experiment. The anti-TT IgG titer and the anti-DT IgG titer increased, and a significant effect was detected by paired t-test. The antibody titers were maintained at higher level than that before vaccination for at least 1 year. Moreover, toxoid-specific antibodies were produced by the second vaccination in some subjects. Antibodies induced by application of the TCI formulation neutralized the toxin and prevented toxic death in mice. In addition, changes in the skin condition due to application of the TCI formulation were observed under in vivo confocal Raman spectroscopy. The amount of water and patch components in the stratum corneum increased after application of the TCI formulation, suggesting that the change in the skin condition was related to antigen penetration. These data indicate that this easy-to-use TCI system induces an immune response without severe adverse reactions in humans. This easy-to-use and safe TCI formulation enables mass treatment in an outbreak setting and increased vaccination rates in developing countries, and will greatly contribute to worldwide countermeasures against infectious diseases.     

Effects of 3-<i>O</i>-Methyldopa, <small>L</small>-3,4-Dihydroxyphenylalanine Metabolite, on Locomotor Activity and Dopamine Turnover in Rats

It has been well known that 3-O-methyldopa (3-OMD) is a metabolite of L-3,4-dihydroxyphenylalanine (L-DOPA) formed by catechol O-methyltransferase (COMT), and 3-OMD blood level often reaches higher than physiological level in Parkinson's disease (PD) patients receiving long term L-DOPA therapy. However, the physiological role of 3-OMD has not been well understood. Therefore, in order to clarify the effects of 3-OMD on physiological function, we examined the behavioral alteration in rats based on locomotor activity, and measured dopamine (DA) and its metabolites levels in rats at the same time after 3-OMD subchronic administration. The study results showed that repeated administrations of 3-OMD increased its blood and the striatum tissue levels in those rats, and decreased locomotor activity in a dose dependent manner. Although 3-OMD subchronic administration showed no significant change in DA level in the striatum, DA metabolite levels, such as 3,4-dihydroxyphenylacetic acid (DOPAC), 3-methoxytyramine (3-MT), and homovanillic acid (HVA) were significantly decreased. After 3-OMD washout period (7?d), locomotor activity and DA turnover in those rats returned to normal levels. Furthermore, locomotor activity and DA turnover decreased by 3-OMD administration were recovered to normal level by acute L-DOPA administration. These results suggested that 3-OMD affect to locomotor activity via DA neuron system. In conclusion, 3-OMD itself may have a disadvantage in PD patients receiving L-DOPA therapy.     

The resistant starch level of heat moisture-treated high amylose cornstarch is much lower when measured in the human terminal ileum than when estimated in vitro

According to the definition of resistant starch (RS), the true value of foodstuff-derived RS can be assessed only from that found in the contents of the terminal ileum. To date, a few methods exist for in vivo measurement of RS in the terminal ileum, but their accuracy is questionable. The aim of this study was to quantify the level of RS in the terminal ileum to determine its true value as dietary fiber (DF). Volunteers (n = 7 men) were given a test meal containing 10 g of heat moisture-treated high amylose cornstarch (HMT-HAS) containing 8.8 g of RS as measured by Englyst's method. A double-lumen tube was positioned in the terminal ileum using the endoscopic retrograde bowel insertion method (ERBI). Intestinal contents were aspirated, and the amount of RS was measured as the glucose concentration (Englyst's method), and compared with the values for RS administrated orally using the same method. The mean amount of HMT-HAS-derived RS collected in the terminal ileum was 3.37 +/- 0.95 g (mean +/- SD), which was 34.5 +/- 9.7% of the in vitro RS value. Furthermore, there were large individual differences in recoveries, ranging from 22.2 to 47.5%. The measured amount of HMT-HAS-derived RS was much smaller in our in vivo study than that measured in vitro, suggesting that in vitro measurement may inaccurately estimate the RS and DF levels of foodstuffs. The problem is further compounded by the large individual in vivo variations in RS values from subjects consuming identical diets.     

Loss of the cyclin-dependent kinase 4-inhibitor (p16; MTS1) gene is frequent in and highly specific to lymphoid tumors in primary human hematopoietic malignancies

The cyclin-dependent kinase 4-inhibitor (CDK41; p16; or MTS1) gene has been proposed as a candidate for a tumor-suppressor gene located in chromosome 9p21, a frequently deleted region in a wide spectrum of human cancers, including leukemias. Recent studies disclosed that it was frequently deleted or mutated in a variety of primary human cancers, including acute lymphoblastic leukemia. The purpose of this study is to figure out the precise manners and frequencies of p16 gene inactivation in diverse hematopoietic tumor types and thus to clarify its significance in development of human hematopoietic malignancies. A total of 410 tumor specimens from patients with primary hematopoietic malignancies were examined for deletions of the p16 gene as well as the neighboring p15 gene and the nearby interferon alpha gene by Southern blot analysis. Tumor-specific mutations or small deletions of the p16 gene were also studied in 74 patients using single-strand conformation polymorphism analysis and direct sequencing. Loss of the p16 gene was most frequently observed among the three genes examined and was found in 59 of the 410 patients: 2 of 134 with acute myelocytic leukemia, 41 of 105 with acute lymphocytic leukemia, 2 of 15 with chronic lymphocytic leukemia, 5 of 14 with adult T-cell leukemia, 4 of 33 with non-Hodgkin's lymphoma, 3 of 8 with mixed-lineage leukemia, and 2 of 61 with chronic myelocytic leukemia. In 16 of the 59 patients, the p16 deletions occurred due to rearrangements within the small region between the p15 exon 2 and the p16 exon 2. Tumor-specific mutations or small deletions of the p16 gene were not detected in the 74 patients examined, including 12 of 14 patients with hemizygous deletions of the gene. Loss of the p16 gene is frequent in and highly specific to lymphoid malignancies (54 of 183 [30%] in lymphoid tumor v2 of 219 [1%] in myeloid tumors; P < .0001). The deletion analyses strongly suggest that the p16 gene is a tumor-suppressor gene located in chromosome 9p21 that is involved in development of human lymphoid tumors. Gene deletions but not minute mutations should be the predominant mechanism of p16 gene inactivation in these types of tumors.     

Ecabet sodium inhibits the ability of Helicobacter pylori to induce neutrophil production of reactive oxygen species and interleukin-8

The pathogenic role of human neutrophils has been implicated in Helicabacter pylori-associated gastritis. Ecabet sodium, a locally acting antiulcer drug, has anti-H. pylori actions. The aim of this study was to examine the effects of ecabet on the ability of H. pylori to stimulate human neutrophils. H. pylori were added to 1 × 10⁵ neutrophils and incubated for 30 min in the presence of ecabet. Bacterial suspensions which had been incubated with ecabet for 30 min were also used to stimulate neutrophils. The intracellular production of reactive oxygen species was measured with a FACScan. Bacterial suspensions were also added to neutrophils in the presence of ecabet and incubated at 37°C for 12 h to measure interleukin (IL)-8 production by enzyme-linked immunosorbent assay. The mean fluorescence intensity was found to be attenuated dose-dependently by ecabet (P < 0.01). Ecabet also inhibited IL-8 production by neutrophils in a dose-dependent manner (P < 0.001). Bacteria with prior incubation with ecabet induced significantly lower IL-8 production than those without this incubation (P < 0.05). Ecabet sodium has preventive effects on the ability of H. pylori to stimulate human neutrophils. It may lead to reduced gastritis activity and decreased oxidative damage of the gastric mucosa in H. pylori-associated gastritis. Recieved: April 10, 2000 / Accepted: September 22, 2000     

Perforated emphysematous cholecystitis managed by endoscopic transpapillary gallbladder drainage

An 88-year-old woman with dementia was diagnosed as having perforated emphysematous cholecystitis with localized peritonitis. Because she was at high risk for surgery, gallbladder drainage was required before surgery. Endoscopic transpapillary gallbladder drainage instead of percutaneous transhepatic biliary drainage was performed because bile could leak from the puncture site to free space around the perforated gallbladder. After the insertion of a nasobiliary drainage tube, the gallbladder was drained and cleaned with saline solution. Subsequently, a nasobiliary drainage tube was replaced with a double-pigtail stent because she was at high risk of dislodging the nasobiliary drainage tube. Although clinical improvement was observed, she was treated conservatively without surgery. She was followed up for 6 months without developing cholecystitis. For perforated cholecystitis without developing panperitonitis, endoscopic transpapillary gallbladder drainage would be an effective option as a bridge to surgery for the initial treatment and as an alternative to surgery for long-term management for a later treatment. This is the first reported case of perforated emphysematous cholecystitis with localized peritonitis treated with endoscopic transpapillary gallbladder drainage.