On the Contamination of Sea Water with Salmonella and Fecal Indicator Organisms : I. Occurrence and Distribution of Salmonella and Fecal Indicator Organisms in Coastal Sea Water of Fukuyama

瀬戸内海の環境汚染調査の一環として1972年6月から1974年3月にわたり,福山市沿岸海水を対象にサルモネラと汚染指標菌の動態について調査した。その結果,以下のような成績が得られた。 1. 8地点(A～H)から採取した海水のサルモネラ陽性率は,平均12.1%(28/232)であった。 2. サルモネラ陽性率は地点および季節によって異なり,A地点23.9%(11/46),B地点21.7%(10/46)およびE地点25%(3/12)と陽性率が高く,また季節別では5～10月に高かった。 3. サルモネラ陽性率は港内から港外および沖合部に行くに従って低下した。 4. 分離された42株のサルモネラの菌型またはO群は,S.typhimurium, S.thompson, S.anatum, S.infantis, S.give, S.schleissheim, S.schwarzengrund, S.derby, S,montevideo, S.meleagridis, S,binza, S.senftenbergおよび型別不能のO群B,C1,D,E1であった。 5. これらの菌型・群は,以前に著者らが当地方の飼料工場や河川から分離した菌型と大多数は同じであったことから,陸上環境を汚染しているサルモネラが最終的には海水まで汚染していることが明らかとなった。 6. 従って海水のサルモネラ汚染は水産物のサルモネラ汚染にも影響を及ぼしている。 7. 汚染指標菌は大腸菌群,大腸菌および腸球菌の順で菌数ならびに検出率が高く,大腸菌群はほとんどが陽性であったが,他の2者では陰性例も多かった。 8. これら汚染指標菌の菌数が多い海水ほど,サルモネラ検出率が高い傾向にあった。 9. サルモネラならびに汚染指標菌の検出状況と海水温との関係を検討した結果,サルモネラは汚染指標菌の菌数が多く,しかも,水温が19℃以上の海水から検出される傾向があった。As a part of the survey on environmental pollution of the Seto Inland Sea, Japan, a survey on the ecology of Salmonella and fecal indicator organisms in the sea water samples collected at eight stations (A to H) located on the coast of Fukuyama, was conducted in June 1972 through March 1974. The results obtained were as follows: 1. The positive rate for occurrence of Salmonella in the samples collected at the eight stations was 12.1% (28/232) in average. 2. It was observed that the frequency of occurrence of Salmonella in the samples varied depending on the sampling stations and on the seasons. Positive rates in Salmonella detection as high as 23.9% (11/46) at St. A, 21.7% (10/46) at St. B, and 25.0 % (3/ 12) at St. E were observed. In the seasonal observations, a higher positive rate of occurrence of Salmonella was observed in the samples collected during May through October. 3. The frequency of incidence of Salmonella in the samples lowered as the sampling station went from the inner part of Fukuyama Port to the outer part of the Port and the offshore area. 4. Some of the forty-two strains of Salmonella isolated were serotyped to S. typhimurium, S. thompson, S. anatum, S. infantis, S. give, S. schleissheim, S. schwarzengrund, S. derby, S. montevideo, S. meleagridis, S. binza and S. senftenberg, and the O-groups B, C1 , D and E 1. 5. Most of the serotypes isolated in the present study were similar to those isolated previously from the river waters and feed-stuffs in this district by the authors. According to the results, it was proved that those serotypes of Salmonella contaminating the terrestrial environment have also contaminated the coastal marine environment as a terminal stage in the pollutive circulation. 6. The marine contamination with Salmonella may result into the contamination of marine products. 7. The higher values of the MPN and frequency of occurrence of the fecal indicator organisms were observed according to the order to coliforms, E. coli and enterococci. The incidence of coliforms was positive in most of samples examined, but those of the others were negative in some cases. 8. A high tendency of Salmonella recovery was observed in cases that represented high values in the MPN of fecal indicator organisms. 9. As a results of consideration on the relationship between the sea water temperature and the frequency of occurrence of Salmonella and of fecal indicator organisms, it was recognized that there is a high tendency of Salmonella recovery in the sea water samples that showed a high MPN value of fecal indicator organisms and were on a temperature higher than 19° C.本研究の一部は昭和47～49年度文部省科学研究費(特定研究1「瀬戸内海の汚染と指標生物の動熊に関する研究―代表者藤山虎也教授」)によるものである

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure < 100 mmHg (n = 1127), estimated glomerular filtration rate < 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

Bone-Orchestrating Cells, Osteocytes

Osteocytes build up functional syncytia, i.e., the osteocytic lacunar-canalicular system（OLCS）. The osteocytes are interconnected through gap junctions between their cytoplasmic processes, which pass through narrow passageways referred to as osteocytic canaliculi. There are two possible ways, in which molecules can be transported throughout the OLCS: via the cytoplasmic processes and their gap junctions, and via the pericellular space in the osteocytic canaliculi. Transport of minerals and small molecules through a spatially well-organized OLCS appears to be pivotal for bone mineral homeostasis and bone remodeling control. Recently, osteocyte-derived molecules -- sclerostin, dentin matrix protein-1, fibroblast growth factor 23（FGF23）-- have been put in evidence as they may be related to osteocytic functions such as regulation of bone remodeling and so forth. Osteocytes were shown to regulate phosphorus serum levels and osteoblastic activity through the expression of FGF23 and sclerostin. In our observations, FGF23 and sclerostin synthesis seemed to be associated with the spatial regularity of the OLCS: both proteins were consistently expressed by osteocytes in epiphyses and cortical bones showing regularly arranged OLCS. In contrast, mice bearing high bone turnover, e.g., osteoprotegerin deficient mice, revealed markedly-diminished sclerostin. This review will introduce our recent studies on the regularity of OLCS and the osteocytic function

Plasma Gel Made of Platelet-Poor Plasma: In Vitro Verification as a Carrier of Polyphosphate

Plasma gel (PG) is a blood-derived biomaterial that can be prepared by heating or chemical cross-linking without the aid of intrinsic coagulation activity and has gradually been applied in the field of esthetic surgery. To explore the applicability of PG in regenerative therapy or tissue engineering, in this study, we focused on the advantages of the heating method and verified the retention capacity of the resulting PG for polyphosphate (polyP), a polyanion that contributes to hemostasis and bone regeneration. Pooled platelet-poor plasma (PPP) was prepared from four healthy male adult donors, mixed with synthetic polyP, and heated at 75 °C for 10 or 30 min to prepare PG in microtubes. The PG was incubated in PBS at 37 °C, and polyP levels in the extra-matrix PBS were determined by the fluorometric method every 24 h. The microstructure of PG was examined using scanning electron microscopy. In the small PG matrices, almost all of the added polyP (~100%) was released within the initial 24 h. In contrast, in the large PG matrices, approximately 50% of the polyP was released within the initial 24 h and thereafter gradually released over time. Owing to its simple chemical structure, linear polyP cannot be theoretically retained in the gel matrices used in this study. However, these findings suggest that thermally prepared PG matrices can be applied as carriers of polyP in tissue engineering and regenerative medicine

Fluorometric Quantification of Human Platelet Polyphosphate Using 4′,6-Diamidine-2-phenylindole Dihydrochloride: Applications in the Japanese Population

Polyphosphate (polyP), a biopolymer of inorganic phosphate, is widely distributed in living organisms. In platelets, polyP is released upon activation and plays important roles in coagulation and tissue regeneration. However, the lack of a specific quantification method has delayed the in-depth study of polyP. The fluorescent dye 4′,6-diamidine-2-phenylindole dihydrochloride (DAPI) has recently received attention as a promising probe for the visualization and quantification of cellular polyP levels. In this study, we further optimized quantification conditions and applied this protocol in quantification of platelet polyP levels in a Japanese population. Blood samples were collected from non-smoking, healthy Japanese subjects (23 males, 23 females). Washed platelets were fixed and probed with DAPI for fluorometric determination. PolyP levels per platelet count were significantly higher in women than that in men. A moderate negative correlation between age and polyP levels was found in women. Responsiveness to CaCl2 stimulation was also significantly higher in women than that in men. Overall, our optimized protocol requires neither purification nor degradation steps, reducing both the time and bias for reproducible quantification. Thus, we suggest that despite its low specificity, this DAPI-based protocol would be useful in routine laboratory testing to quantify platelet polyP levels efficiently and economically

Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control

Although platelet-rich plasma (PRP) is now widely used in regenerative medicine and dentistry, contradictory clinical outcomes have often been obtained. To minimize such differences and to obtain high quality evidence from clinical studies, the PRP preparation protocol needs to be standardized. In addition, emphasis must be placed on quality control. Following our previous spectrophotometric method of platelet counting, in this study, another simple and convenient spectrophotometric method to determine platelet aggregation activity has been developed. Citrated blood samples were collected from healthy donors and used. After centrifugation twice, platelets were suspended in phosphate buffered saline (PBS) and adenosine diphosphate (ADP)-induced aggregation was determined using a spectrophotometer at 615 nm. For validation, platelets pretreated with aspirin, an antiplatelet agent, or hydrogen peroxide (H2O2), an oxidative stress-inducing agent, were also analyzed. Optimal platelet concentration, assay buffer solution, and representative time point for determination of aggregation were found to be 50–100 × 104/μL, PBS, and 3 min after stimulation, respectively. Suppressed or injured platelets showed a significantly lower aggregation response to ADP. Therefore, it suggests that this spectrophotometric method may be useful in quick chair-side evaluation of individual PRP quality