Real-world outcomes of ponatinib treatment in 724 patients with CML and Ph+ ALL: a post-marketing surveillance study with a special interest in arterial occlusive events in Japan.

BACKGROUND: In September 2016, ponatinib was approved in Japan for the treatment of patients with chronic myeloid leukemia with resistance/intolerance to prior tyrosine kinase inhibitors and patients with relapsed or refractory Philadelphia chromosome-positive acute lymphoblastic leukemia. METHODS: We conducted a post-marketing all-case surveillance to study the safety and efficacy of ponatinib in clinical practice, focusing on arterial occlusive events. RESULTS: Data from 724 patients were collected for 2 years from the initiation of ponatinib. The arterial occlusive events were reported in 6.49% (47/724) with an exposure-adjusted incidence rate of 6.8/100 person-years. The risks associated with arterial occlusive events were age and comorbidities including hypertension and diabetes. At 104 weeks, the cumulative major molecular response rate in patients with chronic-phase chronic myeloid leukemia was 67.2% and the complete cytogenetic response in patients with Philadelphia chromosome-positive acute lymphoblastic leukemia was 80.0%. Furthermore, the estimated 1-year overall survival rate was 98.5% for chronic-phase chronic myeloid leukemia and 68.6% for Philadelphia chromosome-positive acute lymphoblastic leukemia. CONCLUSIONS: This surveillance demonstrated that ponatinib has a favorable safety and efficacy profile in Japanese patients and also showed the necessity of closely monitoring arterial occlusive events in older adults and patients with predisposing factors for atherosclerosis.

Phase I study of OPB-51602, an oral inhibitor of signal transducer and activator of transcription 3, in patients with relapsed/refractory hematological malignancies.

We carried out a multicenter dose-escalation phase I study of oral OPB-51602, a signal transducer and activator of transcription 3 phosphorylation inhibitor, in patients with relapsed or refractory hematological malignancies to evaluate the safety, maximum tolerated dose (MTD), pharmacokinetics, and preliminary antitumor activity. Twenty patients were treated with OPB-51602 at doses of 1, 2, 3, 4, and 6 mg in the "3 + 3" dose escalation design. The most common treatment-related adverse events included nausea (55%), peripheral sensory neuropathy (45%), and diarrhea (40%). The most frequently observed grade 3 or 4 drug-related adverse events were neutropenia (20%), leukopenia (15%), lymphopenia (10%), and thrombocytopenia (10%). The MTD was 6 mg, with dose-limiting toxicities of grade 3 lactic acidosis and increased blood lactic acid levels observed in one of three patients and grade 1-2 peripheral neuropathy in three of three patients. The recommended dose was determined to be 4 mg. OPB-51602 was rapidly absorbed, and exposure tended to increase in a dose-dependent manner. Accumulation of OPB-51602 was seen with 4 weeks of multiple treatments. No clear therapeutic response was observed. Durable stable disease was observed in two patients with acute myeloid leukemia and one with myeloma. In conclusion, the MTD of OPB-51602 was 6 mg. OPB-51602 was safe and well tolerated in a dose range of 1-4 mg. However, long-term administration at higher doses was difficult with the daily dosing schedule, and no response was seen. Therefore, further clinical development of OPB-51602 for hematological malignancies with a daily dosing schedule was terminated.

Effects of adiponectin on growth and differentiation of human keratinocytes--implication of impaired wound healing in diabetes.

Impaired wound healing is one of the most common complications associated with diabetes. Adiponectin is an abundant circulating adipocyte-derived cytokine that has beneficial effects on disorders accompanying diabetes. Herein we report that adiponectin has a regulatory effect on the growth and differentiation of HaCaT human keratinocyte cells. Furthermore, adiponectin regulated the expression of TGFbeta isoforms in keratinocytes in a dose-dependent manner, which implies that adiponectin modulates other types of cells related to wound repair via secretion of growth factors from keratinocytes. Moreover, TUNEL assay results revealed that adiponectin enhances apoptosis of keratinocytes. Taken together, the present results indicate that adiponectin has suppressive effects on the formation of hyperkeratosis commonly seen in patients with diabetic foot and its application may be beneficial for developing treatment for that disorder.

A novel method for rapid detection of Streptococcus pneumoniae antigen in sputum and its application in adult respiratory tract infections.

A highly sensitive immunochromatography test kit, ODK0501, was developed using specific polyclonal antibodies against the C-polysaccharide moiety of Streptococcus pneumoniae for the rapid detection of S. pneumoniae antigen in sputum samples. The clinical utility of ODK0501 for this detection was evaluated prospectively in 52 adult patients with respiratory infections and compared with that of a urinary antigen detection kit. Overall, 21 patients (40.4 %) showed positive results with ODK0501, compared with 16 patients (30.8 %) using the urinary antigen detection kit, and S. pneumoniae was cultured from 18 patients. ODK0501 and the urinary antigen detection kit exhibited a sensitivity of 94.4 and 55.6 % (P<0.01), respectively, and a specificity of 88.2 and 82.4 %, respectively. Eleven of thirteen patients with conflicting results between the two test kits exhibited consistent results for sputum cultures. Moreover, eight out of nine patients positive for ODK0501 and negative for the urinary antigen detection kit were S. pneumoniae culture-positive, including five who exhibited phagocytosis, indicating S. pneumoniae as a causative agent of infection, in Gram staining of sputum samples. These results suggest that the ODK0501 direct sputum detection kit is more clinically useful than the urinary antigen detection kit in adult patients with respiratory infections.

Microbacterium terricolae sp. nov., isolated from soil in Japan.

The taxonomic positions of two novel strains isolated from a soil sample collected in Japan using Glucose-Peptone-Meat extract (GPM) agar plates supplemented with superoxide dismutase or superoxide dismutase plus catalase were investigated based on the results of chemotaxonomic, phenotypic and genotypic characteristics. Strains were Gram-positive, catalase-positive, non-motile bacteria with L-ornithine as a diagnostic diamino acid of the peptidoglycan. The acyl type of the peptidoglycan was N-glycolyl. The major menaquinones were MK-12 and 13. Mycolic acids were not detected. The G+C content of the DNA was 70 mol%. Comparative 16S rRNA studies on the two isolated strains revealed that they belong to the genus Microbacterium. DNA-DNA relatedness data revealed that KV-448(T) and KV-769 are a new species of the genus Microbacterium. From these results, we propose that these bacteria should be classified in the genus Microbacterium as Microbacterium terricolae sp. nov. The type strain of Microbacterium terricolae is KV-448(T) (=NRRL B-24468(T), NBRC 101801(T)).

Janibacter corallicola sp. nov., isolated from coral in Palau.

A novel Janibacter species is described on the basis of phenotypic, chemotaxonomic and genotypic data. Two bacterial strains were isolated in Palau, which were both Gram-positive, catalase-positive bacteria with meso-diaminopimelic acid as the diagnostic diamino acid of the peptidoglycan. The major menaquinone was MK-8(H(4)). Mycolic acids were not detected. The G+C content of the DNA was 70-71 mol%. Comparative 16S rDNA studies of the two isolated strains revealed that they both belonged to the genus Janibacter. DNA-DNA relatedness data revealed that 04PA2-Co5-61(T) and 02PA-Ca-009 belong to the same species, a new species of the genus Janibacter. From these results, Janibacter corallicola sp. nov. is proposed, with the type strain 04PA2-Co5-61(T) (=MBIC 08265(T), DSM 18906(T)).

[Taxonomy and phylogenetic analysis of infectious Nocardia strains isolated from clinical samples].

Following recent advances in medical technology, the increased number of immunocompromised patients such as those with organ transplants has led to an increase in opportunistic infections due to Nocardia. Although nocardiosis has been considered to be rare, recent reports indicate that the incidence of the infection is increasing. The Nocardia asteroides group is the principal source of infectious species, but the definition of species in Nocardia is problematic because species identification of N. asteroides has been based on mainly morphological and biochemical properties. Additionally, it was assumed that a clinical strain with properties that did not fully coincide with existing species was N. asteroides sensu lato. This caused problems in both clinical and taxonomical fields, and reclassification of N. asteroides sensu lato was necessary. Therefore, determination of the appropriate taxonomic position of N. asteroides sensu lato that is now classified as N. asteroides sensu stricto was conducted using a molecular phylogenetic method. From 1965 to 2001, twenty-two strains of N. asteroides sensu lato were isolated from clinical samples. The phylogenetic tree using 16S rDNA sequences and detailed biochemical characters on the 22 isolates was determined. Results revealed that nine strains should be reclassified into species other than N. asteroides sensu stricto, and we proposed nine new species of the genus Nocardia. In addition, we proposed eight other new species of the genus Nocardia from other samples; hence seventeen new species were proposed in total. We also reported the first infectious cases due to Nocardia beijingensis, Nocardia transvalensis and Nocardia pseudobrasiliensis in Japan.

First clinical isolates of Nocardia carnea, Nocardia elegans, Nocardia paucivorans, Nocardia puris and Nocardia takedensis in Japan.

Five aerobic actinomycete strains isolated from patients in Japan were assigned provisionally to the genus Nocardia based on morphological and physiological characteristics. The five strains, IFM 10481, IFM 0668, IFM 0901, IFM 0583 and IFM 0342, were not classified into any Nocardia species reported as infectious agents in Japan. Therefore, they were studied further to determine their specific taxonomic positions. Detailed chemotaxonomic and physiologic characterization and 16S rDNA sequence data of the five strains showed that they belonged to respective species of Nocardia carnea, N. elegans, N. paucivorans, N. puris and N. takedensis. This is the first isolation report of these five Nocardia species from patients in Japan.

Nocardia anaemiae sp. nov. isolated from an immunocompromised patient and the first isolation report of Nocardia vinacea from humans.

Two actinomycete strains that were isolated from patients in Japan were assigned provisionally to the genus Nocardia based on their morphological characteristics. The two isolates were further studied to determine their specific taxonomic status. Detailed chemotaxonomic characterization and 16S rDNA sequence data for the strains showed that they are most similar to Nocardia vinacea. DNA-DNA hybridization experiments indicated that strain IFM 0344 should be identified as N. vinacea, and that strain IFM 0323T is classifiable as a new species. This report describes the first isolation of N. vinacea from clinical samples. A new species of the genus Nocardia is proposed based on their phenotypic and phylogenetic characteristics: Nocardia anaemiae for IFM 0323T (=NBRC 100462T=JCM 12396T=DSM 44821T).

Phosphatidylinositol 3-kinase/Akt-dependent and -independent protection against apoptosis in normal human melanocytes.

Normal human melanocytes require the synergistic action of several growth-promoting agents for their growth in serum-free medium. The ability of four representative growth promoting agents including insulin, 12-O-tetradecanoylphorbol-13-acetate (TPA), basic fibroblast growth factor (bFGF), and 3-isobutyl-1-methylxanthine (IBMX), (iTbI) to protect melanocytes against apoptosis was examined. Also, the involvement of phosphatidylinositol (PI) 3-kinase and Akt, one of the downstream targets of PI 3-kinase, in the survival signaling pathway was examined. The percentage of apoptotic cells was negligible when the cells were grown in the presence of iTbI. Deprivation of iTbI from the culture medium for 72 h caused approximately 30% of melanocytes to undergo apoptosis and this was suppressed to variable extents by the addition of one of the iTbI to the medium. Insulin and TPA protected against apoptosis almost completely, whereas bFGF and IBMX rescued melanocytes from apoptosis to a lesser extent. Wortmannin, an inhibitor of PI 3-kinase, potently inhibited the protective effect of insulin on melanocytes, whereas it did not block the ability of TPA, bFGF, or IBMX to rescue the cells from apoptosis. Furthermore, apoptosis of melanocytes induced by deprivation of iTbI was prevented almost completely by infection with an adenovirus vector encoding a constitutively active mutant of either PI 3-kinase or Akt. These results indicate that melanocytes can operate both PI 3-kinase/Akt-dependent and -independent mechanisms for protection against apoptosis and that activation of the PI 3-kinase/Akt pathway is sufficient for protection against apoptosis induced by deprivation of growth-promoting agents.

First isolates of Nocardia abscessus from humans and soil in Japan.

Nocardia abscessus, a recently established species, was isolated from patients during 2000. In the course of our taxonomic studies on 121 clinical Nocardia isolates in Japan 5 strains isolated from patients plus one strain isolated from soil in Japan, were found to have similar physiological characteristics to those of N. abscessus. Phylogenetic studies using 16S rRNA gene sequence analysis confirmed that these strains belong to N. abscessus. This is the first isolation report of N. abscessus from soil as well as from clinical samples in Japan.

Evaluation of a rapid immunochromatographic ODK0501 assay for detecting Streptococcus pneumoniae antigen in sputum samples from patients with lower respiratory tract infection.

A novel, rapid, and noninvasive test (ODK0501) to detect Streptococcus pneumoniae antigen was evaluated in a Japanese multicenter study. ODK0501 uses polyclonal antibodies to detect C polysaccharide of S. pneumoniae from sputum samples by an immunochromatographic assay. The utility of ODK0501 was evaluated for 161 adult patients with lower respiratory tract infection between March 2006 and March 2007. Bacterial culture and identification, real-time PCR, and ODK0501 assays were performed on sputum samples, and the Binax Now Streptococcus pneumoniae antigen test was performed using urine samples obtained from the same patients. The performances of all tests were compared based on the results of bacterial culture and identification. The sensitivity and specificity of ODK0501 were 89.1% (49/55 samples) and 95.3% (101/106 samples), respectively. We then compared the Binax Now Streptococcus pneumoniae antigen test with ODK0501 using samples from 142 patients. The sensitivities of ODK0501 and the Binax Now S. pneumoniae antigen test were 90.0% (45/50 samples) and 62.0% (31/50 samples), respectively (P = 0.002). The relative quantity of S. pneumoniae in expectorated sputum was calculated using real-time PCR and indicated that the possibility of false-positive results for ODK0501 due to indigenous S. pneumoniae was low. The positive and negative concordance rates of ODK0501 and Binax Now were 96.8% (30/31 samples) and 21.1% (4/19 samples), respectively. Binax Now was less capable of detecting S. pneumoniae antigen among patients with underlying chronic obstructive pulmonary disease. In conclusion, ODK0501 is noninvasive, rapid, and an accurate tool for diagnosing respiratory infection caused by S. pneumoniae.

Arthrobacter oryzae sp. nov. and Arthrobacter humicola sp. nov.

Two novel bacterial strains were isolated from a paddy soil sample collected in Japan using GPM agar plates supplemented with superoxide dismutase and/or catalase. The strains were Gram-positive, catalase-positive and motile, with lysine as the peptidoglycan diagnostic diamino acid and acetyl as the peptidoglycan acyl type. The major menaquinone was MK-9(H(2)). Mycolic acids were not detected. The G+C content of the DNA was 66-68 mol%. On the basis of phenotypic analysis, 16S rRNA gene sequence comparisons and DNA-DNA hybridization data, it is proposed that these strains represent two novel species, Arthrobacter oryzae sp. nov. (type strain is KV-651(T)=NRRL B-24478(T)=NBRC 102055(T)) and Arthrobacter humicola sp. nov. (type strain is KV-653(T)=NRRL B-24479(T)=NBRC 102056(T)), respectively.

Humibacillus xanthopallidus gen. nov., sp. nov.

Strain KV-663(T) was isolated from a soil sample collected from a paddy field in Japan by using GPM agar plates. Strain YM21-029 was obtained from a lake sediment sample by using HSV agar. The two novel strains were Gram-positive, catalase-positive, rod-shaped bacteria with ll-diaminopimelic acid as the diagnostic diamino acid of the cell-wall peptidoglycan. The major menaquinone was MK-8(H(4)). Mycolic acids were not detected. The G+C content of the DNA was 69-70 mol%. 16S rRNA gene sequence analysis revealed that these strains represent a novel lineage within the family Intrasporangiaceae, order Actinomycetales, and are related to members of the genera Intrasporangium and Terracoccus. Based on morphological, biochemical and chemotaxonomic properties, together with phylogenetic analysis based on 16S rRNA gene sequences, the two new strains are considered to represent a novel species of a new genus, for which the name Humibacillus xanthopallidus gen. nov., sp. nov. is proposed. The type strain of the type species, Humibacillus xanthopallidus, is KV-663(T) (=NRRL B-24471(T)=NBRC 101803(T)).

Marihabitans asiaticum gen. nov., sp. nov., a meso-diaminopimelic acid-containing member of the family Intrasporangiaceae.

Strain HG667(T), isolated from surface seawater collected at the Kesennuma ferry port in Miyagi Prefecture, Japan, was found to be a Gram-positive, catalase-positive bacterium comprising irregular short rods and cocci. The diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. The major menaquinone was MK-8(H4). Mycolic acids were not detected. The G+C content of the DNA was 70 mol%. Analysis of the 16S rRNA gene sequence revealed that the strain represents a novel lineage within the family Intrasporangiaceae, order Actinomycetales, being associated with the genus Kribbia. On the basis of morphological, biochemical and chemotaxonomic properties of the strain, together with phylogenetic data relating to the 16S rRNA gene sequence, HG667(T) represents a novel genus and species in the family Intrasporangiaceae, for which the name Marihabitans asiaticum gen. nov., sp. nov. is proposed. The type strain of Marihabitans asiaticum is HG667(T) (=MBIC07497(T) =DSM 18935(T)).

Humihabitans oryzae gen. nov., sp. nov.

Strain KV-657(T) was isolated from a paddy field soil sample collected in Japan using GPM agar plates supplemented with catalase. The strain was a Gram-positive, aerobic organism that formed branching hyphae with ll-diaminopimelic acid as the diagnostic peptidoglycan diamino acid. The major menaquinone was MK-8(H(4)). Mycolic acids were not detected. The G+C content of the DNA was 70 mol%. 16S rRNA gene sequence analysis revealed that this strain is closely related to Intrasporangium calvum DSM 43043(T), with a similarity of 97.6 %. Based on the morphological, biochemical and chemotaxonomic properties of this strain and phylogenetic analysis, it was concluded that this isolate represents a new genus and species in the family Intrasporangiaceae, for which the name Humihabitans oryzae gen. nov., sp. nov. is proposed. The type strain of Humihabitans oryzae is KV-657(T) (=NRRL B-24470(T) =NBRC 101802(T)).

Microbacterium sediminicola sp. nov. and Microbacterium marinilacus sp. nov., isolated from marine environments.

Two novel Microbacterium species are described on the basis of phenotypic, chemotaxonomic and genotypic studies. The two strains, designated YM10-847(T) and YM11-607(T), were isolated from river sediment and unidentified hydroid, respectively, of a marine lake. The strains were Gram-positive, catalase-positive bacteria with l-ornithine as the diagnostic diamino acid of the peptidoglycan. The acyl type of the peptidoglycan was N-glycolyl. The major menaquinones were MK-10 and MK-11 for YM10-847(T), and MK-11 and MK-12 for YM11-607(T). Mycolic acids were not detected. The DNA G+C content of strains YM10-847(T) and YM11-607(T) was 67.8 and 71.6 mol%, respectively. Comparative 16S rRNA gene sequence analysis revealed that the two strains belong to the genus Microbacterium. DNA-DNA relatedness data showed that YM10-847(T) and YM11-607(T) are two novel species of this genus. On the basis of these results, strains YM10-847(T) and YM11-607(T) represent two novel species of the genus Microbacterium, for which the names Microbacterium sediminicola sp. nov. and Microbacterium marinilacus sp. nov. are proposed. The type strains are YM10-847(T) (=MBIC08264(T)=DSM 18905(T)) and YM11-607(T) (=MBIC07778(T)=DSM 18904(T)), respectively.

Microbacterium deminutum sp. nov., Microbacterium pumilum sp. nov. and Microbacterium aoyamense sp. nov.

Three novel bacterial strains were isolated from a soil sample collected in Japan by culture on a GPM agar plate supplemented with superoxide dismutase and catalase. The strains were Gram-positive, catalase-positive, non-motile bacteria with L-ornithine as a diagnostic diamino acid of the peptidoglycan. The acyl type of the peptidoglycan was N-glycolyl. The major menaquinones were MK-12, 13 and 14. Mycolic acids were not detected. G+C contents of the DNA were in the range 69-71 mol%. Comparative 16S rRNA gene sequence analysis revealed that the isolates belonged to the genus Microbacterium and were closely related to Microbacterium terregens, Microbacterium aurum, Microbacterium koreense, Microbacterium schleiferi and Microbacterium lacticum. However, M. aurum, M. koreense and M. lacticum clearly differed from the isolated strains based on the presence of L-lysine as the cell-wall diamino acid and various other chemotaxonomic characteristics. Levels of DNA-DNA relatedness showed that the isolated strains represented three separate genomic species. Based on both phenotypic and genotypic data, the following novel species of the genus Microbacterium are proposed: Microbacterium deminutum sp. nov. (type strain KV-483(T)=NRRL B-24453(T)=NBRC 101278(T)), Microbacterium pumilum sp. nov. (type strain KV-488(T)=NRRL B-24452(T)=NBRC 101279(T)) and Microbacterium aoyamense sp. nov. (type strain KV-492(T)=NRRL B-24451(T)=NBRC 101280(T)).

Gordonia araii sp. nov. and Gordonia effusa sp. nov., isolated from patients in Japan.

Two bacterial strains, IFM 10211(T) and IFM 10200(T), were isolated from the sputum of two Japanese patients, and were subjected to a polyphasic taxonomic study. The two strains were found to have morphological, physiological and chemotaxonomic properties that were consistent with their assignment to the genus Gordonia, except for a few chemotaxonomic characteristics. Almost complete 16S rRNA gene sequences of the two strains were determined; the data showed that they are related distantly to Gordonia amarae, Gordonia hirsuta, Gordonia hydrophobica and Gordonia sihwensis, showing 16S rRNA gene sequence similarities to the type strains of these species of 96.2-97.9 %. DNA-DNA relatedness data coupled with the combination of genotypic and phenotypic data indicated that the two strains are representatives of two novel, separate species. The names proposed to accommodate these two strains are Gordonia araii sp. nov. (type strain IFM 10211(T)=DSM 44811(T)=NBRC 100433(T)=JCM 12131(T)) and Gordonia effusa sp. nov. (type strain IFM 10200(T)=DSM 44810(T)=NBRC 100432(T)=JCM 12130(T)).

Nocardia exalbida sp. nov., isolated from Japanese patients with nocardiosis.

Two bacterial strains isolated from different hospitals in Japan were subjected to a polyphasic analysis. Strains IFM 0803(T) and IFM 10383 were found to have morphological, biochemical, physiological and chemotaxonomic properties consistent with their classification in the genus Nocardia. Strains IFM 0803(T) and IFM 10383 clustered with the type strain of Nocardia xishanensis, showing 16S rRNA gene sequence similarities of 98.6-98.9 % with this species. The novel strains could be distinguished from N. xishanensis by a range of phenotypic properties. Based on their phenotypic and phylogenetic characteristics, the two isolates are proposed as members of a novel species of the genus Nocardia, Nocardia exalbida sp. nov., with the type strain IFM 0803(T) (=NBRC 100660(T) = JCM 12667(T) = DSM 44883(T)).