Nagoya Journal of Medical Science

Mammalian cells are frequently exposed to a variety of environmental stresses, such as ultraviolet rays, ionizing radiation, genotoxins, heat shock, and oxidative stress. In coping with the barrage of these and other stresses, multi-cellular eukaryotic organisms have developed a strategy as to how damaged cells will respond to stresses. In general, if the intensity of the damage is moderate, the cell will seek to repair the damage. If, however, the damage to a cell is too severe to be repaired, the affected cells are eliminated by apoptosis. This cell death reduces the risk to the organism as a whole, such as development of a cancer. Such a crucial decision between survival and death is, at least in part, mediated by the stress-activated MAP kinase (SAPK) pathways. SAPKs are a group of serine/threonine protein kinases that convert extracellular stress stimuli into diverse cellular responses, including cell cycle arrest, apoptotic cell death, and cytokine production, through phosphorylation of specific target proteins. Recent progress in the identification of molecules that participate in the SAPK pathways, such as GADD45 proteins and Wip1, has provided new insights, not only into the molecular basis of the cellular response to environmental stress, but also into the etiology of human diseases including cancer.

Recently, liquid-based cytology (LBC) has been widely applied to various samples in diagnostic cytology and its usefulness has been reported. In this study, we investigated thyroid cytology that applied LBC and immunocytochemistry to achieve more objective diagnosis and greater diagnostic accuracy. This study included 125 cases (57 papillary carcinomas (PCs), 22 follicular tumors, 43 adenomatous goiters and 3 with Basedow’s disease). After preparing the LBC slide, immunocytochemical staining was performed on each slide with six antibodies (HBME-1, cytokeratin 19 (CK19), high molecular weight cytokeratin (34βE12), galectin-3, CD15 and CA 19-9). All antibodies presented immunopositivity frequently in PCs, but only a few or some of them were positive in other cases. These antibodies were considered positive markers for PCs, and the most reliable marker was 34βE12; its sensitivity, specificity and diagnostic accuracy were 82.5%, 100% and 92.0%, respectively. Relations of immunocytochemical profiles against these markers were assessed using panel 34βE12, GAL-3 and CK19. More than or equal to two of these markers showed co-positive in 53 of 57 PCs, and negative for all markers was observed in only one case. In the other (non PC) cases, the former was 0 of 58 and the latter was 40 cases. In this panel, the sensitivity, specificity and diagnostic accuracy were 93.0%, 100% and 96.8%, respectively. All of these values were higher than or equal to single values of 34βE12. We concluded that the panel in this study is useful for more objective and accurate diagnosis of thyroid cytology.

Lansoprazole (LPZ), amoxicillin (AMPC) and clarithromycin (CAM) are commonly used drugs (LAC regimen) for Helicobacter pylori (H. pylori) eradication, but the eradication rate with this regimen was reported to be 70% to 90%. A few studies have reported that a successful eradication was associated with the CYP2C19 genotype, which influences the metabolism of proton pump inhibitors (PPI) including LPZ. This study examined the changes in the H. pylori eradication rates between the periods before and after the commencement of a routine genetic test for CYP2C19 at the Daiko Medical Center in Nagoya, Japan, in November, 2005. Subjects were patients who visited the Center during the period from June, 2004 to August, 2010. The patients were classified into three groups according to their CYP2C19 genotype: rapid metabolizers (RM) with a *1*1 genotype, intermediate metabolizers (IM) with a *1*2 or *1*3 genotype, and poor metabolizers (PM) with a *2*2, *2*3, or *3*3 genotype. Non-rapid metabolizers (IM and PM) were basically treated with a LAC regimen, while RMs were treated with a RAM regimen (rabeprazole, AMPC, and metronidazole). The eradication rate was 80.0% (n=90) for the period without the genetic testing and 88.7% (n=124) for the period with the genetic testing (χ2=3.11, p=0.078). The age-sex adjusted odds ratio of eradication success was 2.29 (95% confidence interval, 0.99–5.28, p=0.051) for the latter period relative to the former period among those less than 70 years of age. Those results suggested that the routine genetic test which allows a choice of the RAM regimen for RM improved the eradication rate.

Bilirubin is glucoronized by uridine diphosphate-glucuronosyl transferase 1A1 (UGT1A1) mainly in the liver, and excreted into bile. The conjugated form is metabolized into the unconjugated form, and then into urobilinogen by bacteria in the intestine. Unconjugated bilirubin and urobilinogen are absorbed into the blood stream. The kidney filtrates conjugated bilurubin and urobilinogen into urine. Accordingly, the reduced enzyme activity of UGT1A1 may decrease serum conjugated bilirubin levels, resulting in a lower frequency of positive results of urine bilirubin and urobilinogen. This study examined the associations of UGT1A1 Gly71Arg (UGT1A1*6) with urine bilirubin and urobilinogen, as well as serum AST, ALT and GGT. Subjects were 5,172 inhabitants 35 to 69 years old who participated in a cohort study in Nagoya from June 2008 to May 2010. Among them, data from 5,151 participants (1,465 males and 3,686 females) were available for analysis. The age-sex-adjusted odds ratio (OR) of ArgArg relative to GlyGly was 1.37 (95% confidence interval (95% CI), 0.55–1.23) for bilirubin, and 1.67 (95% CI, 0.86–3.26) for urobilinogen. Those of ArgArg+ArgGly were 0.87 (95% CI, 0.59–1.27) and 1.50 (95% CI, 1.17–1.94), respectively. AST, ALT and GGT levels had no associations with the genotype. Although the significant association for urobilinogen was contrary to the biological expectation, this study indicated that UGT1A1 Gly71Arg may be a genetic factor of urine urobilinogen.

Although the SLC22A12 (uric acid transporter 1) 258X allele is known to cause hypouricemia, the genotype frequency according to the serum uric acid (SUA) level has not been reported. This study investigated the SLC22A12 258WX frequency according to SUA levels among Japanese health-checkup examinees. In addition, the changes were reported in SUA levels during five years for individuals with 258WX. Subjects were 746 Japanese aged 39–86 years in 2003. Their SUA records were linked during the five years from 2003 to 2007. SLC22A12 W258X was genotyped using a polymerase chain reaction with confronting two-pair primers. The 258X allele comprised 1.9% (95% CI, 1.3–2.8%) of all the subjects. Among those with SUA <3.0 mg/dL, 258WX was more common in males (66.7%, 95% CI, 22.2–95.7%) than in females (39.3%, 95% CI, 21.5–59.4%). Among subjects with a SUA of 3.0–4.9 mg/dL, those with 258WX totaled 10.7% (95% CI, 4.0–21.9%) and 2.6% (95% CI, 1.1–5.0%), respectively. There were no subjects with 258WX among those with a SUA of 5.0 mg/dL or more. During the five years from 2003 to 2007, the changes in SUA among 23 individuals with 258WX were found to be similar to those among 258WW subjects (n=536). This study indicated that SLC22A12 258WX was more common among those with a lower serum uric acid concentration. The observed SUA level changes in individuals with 258WX suggested that lifestyle factors could influence the levels of those with 258WX.

Here we show that bis(3’–5’) cyclic diadenylic acid (c-di-AMP) and a diadenylate cyclase (DAC) domain protein involved in the biosynthesis of c-di-AMP were identified in Streptococcus pyogenes. The matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrum of the cell extract of S. pyogenes, which showed a fragment pattern very similar to that of the authentic sample of c-di-AMP, revealed that S. pyogenes produces c-di-AMP in the cell. Subsequently, we confirmed by an in vitro experiment that the production of c-di-AMP in the cell is due to the action of Spy1036 gene encoding a DAC domain protein named spyDAC, which is a new protein different from a well-known diadenylate cyclase. Moreover, the experiment gave a product with a molecular weight of 657.021, which is consistent with the molecular weight of c-di-AMP. Furthermore, the mass spectral fragment pattern of the product obtained by the in vitro biosynthesis is quite similar to that of the product produced by the above in vivo experiment. This in vitro production of c-di-AMP indicated that spyDAC in S. pyogenes actually catalyzes the in vivo biosynthesis of c-di-AMP from ATP.

Public health centers (PHCs, hokenjo in Japanese) are local government authorities responsible for public health in Japan. As of 2010, 494 centers were operating under the Ministry of Health, Labour and Welfare of Japan. While the general rule is that one PHC covers 300,000 inhabitants, several centers cover a population of more than 1 million. The roles of PHCs described in the Community Health Act include 1) propagation and improvement of information on community health, 2) vital statistics and other statistics on community health, 3) improvement of dietary conditions and food sanitation, 4) environmental sanitation including housing, water supply, sewage, waste disposal and public cleaning, 5) medical and pharmaceutical affairs, 6) matters involving public health nurses, 7) promotion and improvement of public medical services, 8) maternal, child, and elderly health, 9) dental health, 10) psychiatric health, 11) health of patients under long-term care due to incurable diseases, 12) prevention of infectious diseases, 13) laboratory tests on sanitation/environment, and 14) other functions needed to maintain/promote health in the community. Among those many roles, infectious disease controls are one of the most important. Concerning tuberculosis control, PHCs are responsible for the isolation of patients, health check-ups of those in close contact with infectious TB patients, and public subsidy of medical expenses for tuberculosis treatments. Food poisoning controls are also an important responsibility of PHCs, as are the conduct of surveys to trace suspicious foods and laboratory testing of samples from patients. To make these many measures effective, sufficient numbers of public health professionals are required.