Potentially inappropriate medications in elderly Japanese patients: effects of pharmacists' assessment and intervention based on Screening Tool of Older Persons' Potentially Inappropriate Prescriptions criteria ver.2.

WHAT IS KNOWN AND OBJECTIVES: The Screening Tool of Older Persons' Potentially Inappropriate Prescriptions (stopp) criteria were updated in 2014 (stopp criteria ver.2), but few studies have evaluated the usefulness of stopp criteria in elderly patients. This prospective observational study evaluated the prevalence of potentially inappropriate medications (PIMs), and the efficacy of hospital pharmacists' assessment and intervention based on stopp criteria ver.2. METHODS: The study was conducted at three medical units of Kobe University Hospital between April 2015 and March 2016. Pharmacists assessed and detected PIMs based on stopp criteria ver.2 and considered the patient's intention to change the prescription at the time of admission of each patient. If the pharmacists judged that benefits outweighed risks of prescription change and the patients consented to change the medications, they recommended the doctor to change the prescription. If there was a risk of exacerbation of disease by the change of medications and the pharmacists judged it to be difficult to adjust medications during hospitalization or the patients did not consent to change the medications, they did not recommend to change it. The pharmacists and the doctors discussed and finally decided whether to change the PIMs or not. The number of patients prescribed PIMs, the number and contents of PIMs, and the number of medications changed after pharmacists' intervention were calculated. RESULTS: Totally, 822 new inpatients aged ≥65 years prescribed ≥1 daily medicine were included. Their median (interquartile range) age was 75·0 (71·0-80·0) years, and 54·9% were male. According to the criteria, 346 patients (42·1%) were prescribed ≥1 PIMs. Patients prescribed PIMs took significantly more medications than others: 10·0 (7·0-13·0) vs. 6·0 (4·0-9·0), P < 0·001. The total number of PIMs was 651%, 47·6% of which (n = 310) were recommended the doctors to change, and 292 of 651 PIMs (44·9%) were finally discontinued/changed after pharmacists' assessment and intervention. PIMs related to benzodiazepines, including Z-drugs, were most frequent, with a detailed classifications as follows (changed/total): (i) benzodiazepines for 4 or more weeks (75/205), (ii) drugs that predictably increase the risk of falls in older people (benzodiazepines) (30/67) and (iii) drugs that predictably increase the risk of falls in older people (hypnotic Z-drugs) (15/31). CONCLUSION: Over 40% elderly patients were prescribed PIMs, and pharmacists' assessments and interventions based on stopp criteria ver.2 were useful in detecting and correcting prescription of PIMs.

Detection of Francisella tularensis and analysis of bacterial growth in ticks in Japan.

UNLABELLED: Francisella tularensis is distributed in the Northern hemisphere and it is the bacterial agent responsible for tularaemia, a zoonotic disease. We collected 4 527 samples of DNA from ticks in Japan, which were then analysed by real-time PCR and nested PCR. Francisella DNA was detected by real-time PCR in 2·15% (45/2 093) of Ixodes ovatus, 0·66% (14/2 107) of I. persulcatus, 8·22% (6/73) of I. monospinosus and 0·72% (1/138) of Haemaphysalis flava specimens. Finally, Francisella DNA was detected by nested PCR in 42 and five samples I. ovatus and I. persulcatus, respectively, which were positive according to real-time PCR. Phylogenetic analysis showed that the sequence from I. ovatus and I. persulcatus were clustered with F. tularensis type B strains distributed in Eurasia. Microinjected live F. tularensis persisted in ticks, whereas heat-killed F. tularensis decreased. Microinjected F. tularensis hlyD mutant decreased in ticks significantly compared to parent strain, thereby suggesting that HlyD in F. tularensis contributes to the adaptation or survive of bacterial infection in ticks. SIGNIFICANCE AND IMPACTS OF THE STUDY: Francisella tularensis has been detected in ticks, suggesting that it is a tick-borne pathogen. However, F. tularensis has not been detected in ticks in Japan since 1991. In this study, we performed a large-scale analysis of DNA isolated from ticks in Japan and detected F. tularensis by real-time polymerase chain reaction (PCR) and nested PCR. We found that F. tularensis could survive in ticks based on an experimental tick-infection model. We also identified a bacterial factor that contributes to survival in ticks. Our results suggest that ticks are candidate vectors that mediate F. tularensis infection in Japan.

Seroprevalence of tularemia in wild bears and hares in Japan.

Tularemia is a zoonotic disease caused by Francisella tularensis. The distribution of the pathogen in Japan has not been studied well. In this study, seroprevalence of tularemia among wild black bears and hares in Japan was determined. Blood samples collected from 431 Japanese black bears (Ursus thibetanus japonicus) and 293 Japanese hares (Lepus brachurus) between 1998 and 2009 were examined for antibodies against F. tularensis by micro-agglutination test (MA) or enzyme-linked immunosorbent assay. By subsequent confirmatory tests using western blot (WB) and indirect immunofluorescence assay (IFA), eight sera from Japanese black bears were definitely shown to be seropositive. All of these eight bears were residents of the northeastern part of main-island of Japan, where human tularemia had been reported. On the other hand, no seropositive Japanese hares were found. These results suggest that Japanese black bears can serve as sentinel for tularemia surveillance and may help understand the distribution of F. tularensis throughout the country. This is the first report on detection of antibody to F. tularensis in black bears of Japan.

Structural and functional analyses of chicken liver ferritin.

Characterization of ferritins from different species has provided insight into iron regulation mechanisms and evolutionary relationships. Here, we examined chicken liver ferritin, which comprises only H subunit and has 14.8 µg of Fe/100 µg of protein. The chicken H subunit apo homopolymer showed the same iron uptake rate as bovine H subunit homopolymer expressed with a baculovirus expression system (0.31 and 0.28 mmol of Fe/min per micromole of protein for chicken and bovine H subunit, respectively). Chicken H subunit apo homopolymer showed a significantly higher biotinylated hemin-binding activity than liver holoferritin. Although bovine spleen apoferritin, which has an L (liver or light):H (heart or heavy) subunit ratio of 1:1, also shows a significantly higher biotinylated hemin-binding activity than its holoferritin, these biotinylated hemin-binding activities were markedly lower than those of both chicken holo- and apoferritins. Binding of chicken holo- and apoferritin with biotinylated hemin was strongly inhibited by hemin but not iron-free hemin, protoporphyrin IX, or Zn-protoporphyrin. These findings demonstrate that chicken ferritin comprises only an H subunit, possesses ferroxidase activity as in mammalian ferritin H subunits, and binds heme more strongly than mammalian ferritins.

Characterization of an immortalized oviduct cell line from the cynomolgus monkey (Macaca fascicularis).

To establish reproductive biological techniques in mammals, it is important to understand the growth environment of the embryo. Oviduct epithelial cells are in close proximity to the embryo during pre-implantation development. We, therefore, established an immortalized oviduct epithelial cell line from the cynomolgus monkey, evaluated the usefulness of these cells as feeder cells for embryo culture, and investigated the gene expression of several growth factors and cytokines in the cells. The immortalized cells were positive for the anti-cytokeratin antibody, as determined by immunocytochemistry, indicating that they are epithelial. They also expressed oviductin, which is specific to oviduct epithelial cells, glyceraldehyde-3-phosphate dehydrogenase (control), leukemia inhibitory factor, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor 1, transforming growth factor beta-2, and interleukin 4. Mouse embryo development was improved when the immortalized cells were used as feeder cells. This cell line is also useful for studying the factors secreted by oviduct epithelial cells.

Identification of an amino acid responsible for the CD3 polymorphism in cynomolgus monkeys (Macaca fascicularis).

The FN18 monoclonal antibody (mAb), directed to CD3 molecules, did not react with the lymphocytes of some cynomolgus monkeys (Macaca fascicularis), because of the polymorphism of the CD3epsilon chain. The epitope recognized by the FN18 mAb was successfully expressed on COS7 cells upon transfection of plasmid DNA coding for the CD3epsilon derived from T cells of a FN18 positive cynomolgus monkey. By construction and expression of plasmid DNA encoding the mutant CD3epsilon, the amino acid residue at position 67 was demonstrated to be involved in the formation of an epitope recognizable by the FN18 mAb.

CD3 polymorphism in cynomolgus monkeys (Macaca fascicularis).

Cynomolgus monkeys were divided into two groups in terms of the reactivity of their lymphocytes with the FN18 monoclonal antibody, which is directed to the CD3 of rhesus monkeys. It was shown that 24 (12.2%) out of 196 monkeys did not have lymphocytes that reacted with the FN18, although T cells from those animals responded well to mitogenic stimulation. We have determined the nucleotide sequences of the CD3delta, CD3gamma, and CD3epsilon chains and found that two amino acids of the CD3epsilon chain of the FN18 non-reactive monkeys were different when compared with the FN18 reactive monkeys. Our results indicated that the CD3epsilon molecule of cynomolgus monkeys is polymorphic at the epitope level, which is recognized by the FN18 monoclonal antibody.

Isolation and characterization of a human alternative complement pathway-inhibiting protein from larval hemolymph of the silkworm, Bombyx mori.

An alternative complement pathway-inhibiting protein (ACPIP), which inhibits the activation of the alternative complement pathway (ACP) of the human serum, was isolated from larval hemolymph of the silkworm, Bombyx mori, by using ammonium sulfate fractionation and column chromatographies to homogeneity. About 400microg of ACPIP was routinely obtained from 20ml hemolymph. The purified ACPIP preparation consisted of two distinct polypeptides (34 and 32kDa) on SDS-PAGE. The amino acid compositions of the two polypeptides were nearly identical; 21% of the amino acid residues were acidic. The amino terminal amino acid sequences up to 20 residues in these two polypeptides were also identical. Addition of the ACPIP to human serum resulted in a dose-dependent inhibition of the hemolysis of intact rabbit erythrocytes via the ACP, whereas in no inhibition of hemolysis of sensitized-sheep erythrocytes (EA) via the classical pathway.

An allosteric drug, o,o'-bismyristoyl thiamine disulfide, suppresses HIV-1 replication through prevention of nuclear translocation of both HIV-1 Tat and NF-kappa B.

The efficacy of o,o'-bismyristoyl thiamine disulfide (BMT) was examined in detail against HIV-1 laboratory isolates (HTLV-IIIB, JRFL, and MN), primary isolates (KMT and KMO), and simian immunodeficiency virus (SIVmac251) in vitro. BMT inhibited the replication of HIV-1 in both laboratory and primary isolates in vitro. In addition, BMT exhibited antiviral activity against SIVmac251. Minimizing energy studies of BMT structure reveal that a trans-disulfide of thiamine (holo drug) disulfide (TDS, protodrug) is allosterically transited to the reactive twisted disulfide of BMT (allo drug) by o,o'-bismyristoyl esterification of TDS. BMT inhibits nuclear translocation of both HIV-1 transactivator (TAT) and the cellular transcriptional nuclear factor-KB (NF-kappa B), resulting in the suppression of HIV-1 replication.

Cephalexin concentrations in radicular granuloma following a single oral administration of 250- or 500-mg cephalexin.

1. Cephalexin concentrations in radicular granuloma and serum following a single oral administration of 250- or 500-mg cephalexin were measured by a paper disk method. 2. The highest concentration of cephalexin in radicular granuloma following administration of 250-mg cephalexin to nonfasting patients was observed at 2 hr, and was 1.62 micrograms/g. The mean cephalexin concentration ratio of radicular granuloma/serum at 2 hr was 0.35. 3. The highest concentrations of cephalexin in radicular granuloma following administration of 500-mg cephalexin to nonfasting and fasting patients occurred at 2 and 1.5 hr, and was 3.35 and 3.42 micrograms/g, respectively. Mean cephalexin concentration ratios of radicular granuloma/serum at 2 and 1.5 hr were 0.32 and 0.30, respectively. 4. All mean cephalexin concentrations in radicular granuloma following administration of 500-mg cephalexin to both fasting and nonfasting patients exceeded MIC for 90% (2 micrograms/ml) of clinically isolated strains of alpha-hemolytic streptococci. However, those concentrations obtained by 250-mg cephalexin did not exceed it.

Amoxicillin concentration in pus from abscess caused by odontogenic infection.

1. Amoxicillin concentration in pus from odontogenic infection was assayed and the concentrations were compared with MIC (minimum inhibitory concentration) of alpha-hemolytic streptococci isolated from odontogenic infection. 2. Measurable amoxicillin concentrations in serum and pus were found in all instances (n = 16). 3. The mean peak concentrations in serum and pus were found at identical times, 1.5 hr after administration, which were 5.92 and 0.90 micrograms/ml, respectively. 4. The mean concentration ratio of pus/serum at the peak time was 0.15. 5. All amoxicillin concentrations in pus at the peak time exceeded the MIC for 90% of alpha-hemolytic streptococci (0.25 micrograms/ml).

Concentrations of lomefloxacin in radicular cyst and oral tissues following single or multiple oral administration.

Concentrations of lomefloxacin in serum, the wall and fluid of radicular cyst, gingiva, and jawbone following single or multiple oral administration were measured. The highest concentrations of lomefloxacin in serum, cyst wall, cyst fluid, gingiva, and jawbone occurred at 3 h after multiple administration, and were 2.31 micrograms/ml, 4.06 micrograms/g, 1.54 micrograms/ml, 4.72 micrograms/g and 2.79 micrograms/g, respectively. The mean concentration ratios of wall/serum, fluid/serum, fluid/wall, gingiva/serum, and jawbone/serum at the highest concentrations were 1.74, 0.73, 0.47, 2.52 and 1.20, respectively. Although most lomefloxacin concentrations in cyst and oral tissues following single oral administration did not exceed the MICs for 80% of clinically isolated strains of alpha-hemolytic streptococci, Staphylococcus aureus and Niesseria spp., most of those obtained after multiple oral administration exceeded the MICs except in the case of fluid.

Ampicillin concentrations in radicular cysts following a single oral administration of bacampicillin.

1. Ampicillin concentrations in cyst wall (wall) and cyst fluid (fluid) of radicular cyst and serum following a single oral administration of bacampicillin (equivalent to 500 mg of ampicillin) were measured by a paper disk method. 2. The mean peak concentrations of ampicillin in wall, fluid, and serum occurred at identical times, 1.5 hr, and were 2.39 micrograms/g, 0.77, and 10.24 micrograms/ml, respectively. 3. Mean ampicillin concentration ratios of wall/serum, fluid/serum, and fluid/wall at the peak time were 0.23, 0.07, and 0.40, respectively. 4. Mean ampicillin concentrations in wall and fluid at the peak time exceeded MIC (minimum inhibitory concentration) for 90% (0.5 microgram/ml) for clinically isolated strains of alpha-hemolytic Streptococci.

Josamycin concentrations in radicular cysts following a single oral administration.

1. Josamycin concentrations in cyst wall (wall) and cyst fluid (fluid) of radicular cyst and serum following a single oral administration of josamycin (600 mg) were assayed by a paper disk method. 2. The mean peak josamycin concentrations in wall, fluid and serum occurred at 1.5, 2 and 1.5 hr, respectively, and were 1.33 micrograms/g, 0.53 and 0.74 micrograms/ml, respectively. 3. The mean concentration ratios of wall/serum, fluid/serum and fluid/wall at the peak times (1.5, 2 and 2 hr) were 1.80, 0.64 and 0.42, respectively. 4. Josamycin concentrations in wall and fluid at the peak time exceeded MIC for 80% for clinically isolated strains of alpha-hemolytic Streptococci.

Cefaclor concentrations in human serum, gingiva, mandibular bone, and dental follicle following a single oral administration.

1. Cefaclor concentrations in human serum (n = 59), gingiva (n = 46), mandibular bone (n = 39), and dental follicle (n = 42) following a single oral administration of cefaclor (500 mg) were measured by the paper disk method. 2. The peak times of serum, gingiva, mandibular bone, and dental follicle were 1.5, 2, 2, and 1.5 hr, respectively. 3. The mean peak concentrations of serum, gingiva, mandibular bone, and dental follicle were 7.58 micrograms/ml, 3.71, 1.59 and 2.42 micrograms/g, respectively. 4. The concentration ratios of gingiva/serum, mandibular bone/serum, and dental follicle/serum at peak times of the tissues were 0.49, 0.18, and 0.32, respectively. 5. Mean cefaclor concentrations in gingiva, mandibular bone, and dental follicle at peak times exceeded MIC for 90% for clinically isolated strains of alpha-hemolytic Streptococci.

Ampicillin concentrations in human oral tissues following a single oral administration of lenampicillin.

1. Ampicillin concentrations in serum (n = 20), gingiva (n = 12), jawbone (n = 13), dental follicle (n = 12), radicular granuloma (n = 2) and radicular cyst (n = 2) were measured in specimens obtained during 0.5-2.5 hr after a single oral administration of lenampicillin (equivalent to 500 mg of ampicillin). 2. Measurable ampicillin concentrations were found in all serum and tissues. 3. Ampicillin concentrations in serum and tissues except for some gingiva and jawbone exceeded MIC for 90% of clinically isolated strains of alpha-hemolytic Streptococci. 4. Ampicillin concentrations in gingiva and jawbone were below the MIC for 90% in 2 out of 12 and 4 out of 13 specimens, respectively.

Concentrations of ampicillin in human serum and mixed saliva following a single oral administration of lenampicillin, and relationship between serum and mixed saliva concentrations.

The concentrations of ampicillin in serum and mixed saliva after a single oral administration of lenampicillin (500 mg) were determined by the paper disc method. The samples of serum and mixed saliva were obtained at 1, 2, 3 and 4 h after administration. The highest concentrations of ampicillin in serum and mixed saliva occurred 1 h after administration of lenampicillin, and were 11.55 and 0.060 micrograms/ml, respectively. A significant correlation coefficient between the concentrations of ampicillin in serum and mixed saliva, r = 0.71, P less than 0.001, was found.

Cephalexin concentrations in human serum, gingiva, and mandibular bone following a single oral administration.

1. Cephalexin concentrations in human serum, gingiva, and mandibular bone after a single oral administration of cephalexin (500 mg) were measured by the paper disc method. 2. The peak times of serum, gingiva, and mandibular bone were approximately 90, 120 and 120 min, respectively. 3. The peak concentrations of serum, gingiva, and mandibular bone were 10.58 micrograms/ml, 5.57 micrograms/g and 2.12 micrograms/g, respectively. 4. The concentration ratio of gingiva/serum and mandibular bone/serum peak time of serum were 0.47 and 0.18, respectively. 5. Cephalexin concentrations in gingiva and mandibular bond did not exceed the MIC80s for clinically isolated strains of Staphylococcus aureus spp., alpha-Streptococci and Peptostreptococcus spp.

Josamycin concentrations in human dental granuloma after a single oral administration of josamycin.

1. Josamycin concentrations in human serum and dental granuloma after a single oral administration of josamycin (600 mg) were assayed by an agar diffusion (paper disc) method. 2. The mean peak josamycin concentrations in serum and dental granuloma occurred at an identical time, approximately 90 min, and were 0.88 micrograms/ml and 1.61 micrograms/g, respectively. 3. The mean concentration ratio of dental granuloma to serum at the peak time was 2.24. 4. Josamycin concentration in dental granuloma at the peak time exceeded MIC80 for clinically isolated strains of Streptococcus group A, Peptostreptococcus spp., and Bacteroides spp.

Ampicillin concentrations in human serum and periodontal membrane following a single oral administration of talampicillin.

1. Ampicillin concentrations in human serum and periodontal membrane after a single oral administration of talampicillin (500 mg) were assayed by the agar diffusion (paper disc) method. 2. The peak times of serum and periodontal membrane were identical, being 150 min after administration. 3. The peak concentrations of serum and periodontal membrane were 7.81 micrograms/ml and 4.11 micrograms/g, respectively. 4. The mean ratio of periodontal membrane to serum concentration at the peak time was 0.53.