Effects of alginate oligomer on the expression of cell cycle- and stress-related genes in Chlamydomonas reinhardtii.

Enzymatically prepared alginate oligomer (AO) promoted the growth of Chlamydomonas reinhardtii in a concentration-dependent manner. AO at 2.5 mg/mL induced increase in expression levels of cyclin A, cyclin B, and cyclin D in C. reinhardtii. CuSO4 at 100 µM suppressed the growth of C. reinhardtiin, and AO at 2.5 mg/mL significantly alleviated the toxicity of CuSO4. Increased intracellular reactive oxygen species level in C. reinhardtii induced by CuSO4 was reduced by AO. After cultivation with CuSO4 at 100 µM, expression levels of ascorbate peroxidase and superoxide dismutase in C. reinhardtii were increased, and AO reduced the increased levels of these enzymes. These results suggest that AO exhibits beneficial effects on C. reinhardtii through influencing the expression of various genes not only at normal growth condition but also under CuSO4 stress.

Alginate oligomer induces nitric oxide (NO) production in RAW264.7 cells: elucidation of the underlying intracellular signaling mechanism.

Alginate is an acidic linear polysaccharide with immune-modulating activities. In this study, we found that enzymatically digested alginate oligomer (AO) with various degrees of polymerization (DP; 2-5) induced a higher level of nitric oxide (NO) production in RAW264.7 cells than undigested alginate polymer (AP). Reverse transcription-polymerase chain reaction and western blot analyses revealed that the expression level of inducible NO synthase in AO-treated RAW264.7 cells was higher than that in AP-treated cells. AO induced nuclear translocation of nuclear factor (NF)-κB p65 subunit in RAW264.7 cells to a greater extent than AP. Although AO and AP induced similar extents of phosphorylation in three mitogen-activated protein (MAP) kinases, c-Jun N-terminal kinase inhibitor exhibited the most potent inhibitory effect on NO induction in AO- and AP-treated RAW264.7 cells, among three MAP kinase inhibitors that were tested.

Heat exposure enhances radiosensitivity by depressing DNA-PK kinase activity during double strand break repair.

PURPOSE: From the role of double strand DNA dependent protein kinase (DNA-PKcs) activity of non-homologous end joining (NHEJ) repair for DNA double strand breaks (DSBs), we aim to define possible associations between thermo-sensitisation and the enzyme activities in X-ray irradiated cells. MATERIALS AND METHODS: DNA-PKcs deficient mouse, Chinese hamster and human cultured cells were compared to the parental wild-type cells. The radiosensitivities, the number of DSBs and DNA-PKcs activities after heat-treatment were measured. RESULTS: Both DNA-PKcs deficient cells and the wild-type cells showed increased radiosensitivities after heat-treatment. The wild-type cells have two repair processes; fast repair and slow repair. In contrast, DNA-PKcs deficient cells have only the slow repair process. The fast repair component apparently disappeared by heat-treatment in the wild-type cells. In both cell types, additional heat exposure enhanced radiosensitivities. Although DNA-PKcs activity was depressed by heat, the inactivated DNA-PKcs activity recovered during an incubation at 37 °C. DSB repair efficiency was dependent on the reactivation of DNA-PKcs activity. CONCLUSION: It was suggested that NHEJ is the major process used to repair X-ray-induced DSBs and utilises DNA-PKcs activity, but homologous recombination repair provides additional secondary levels of DSB repair. The thermo-sensitisation in X-ray-irradiated cells depends on the inhibition of NHEJ repair through the depression of DNA-PKcs activities.

Renal artery intervention using a virtual 4.5-Fr guiding catheter.

A 6.5-Fr guiding catheter (Parent Plus 45, Medikit, Tokyo, Japan), composed of a hydrophilic sheathless guiding catheter and a dilator, has recently been developed for the treatment of renal artery stenosis. Because of its unique sheathless configuration, its outer diameter is comparable to that of a conventional 4.5-Fr introducer sheath, with the inner diameter being close in size to a 6.5-Fr guiding catheter. These features allow this sheathless system to be used as the sole guiding catheter while the arterial damage at the puncture site remains equivalent to that of a 4.5-Fr introducer sheath. We thus call this novel sheathless guiding catheter a virtual 4.5-Fr system. To demonstrate the potential of this virtual 4.5-Fr system, we report a case of renal artery stenosis successfully treated via the transradial route. This virtual 4.5-Fr system may become a viable alternative to conventional guiding catheters, and provide a favorable impact upon vascular access complications and patient morbidity.

Risk of long-term dual antiplatelet therapy following drug-eluting stent implantation in octogenarians.

OBJECTIVES: To evaluate the risk of long-term dual antiplatelet therapy (DAT) following drug-eluting stent (DES) implantation in octogenarians. BACKGROUND: DES implantation requires DAT; however, DAT-associated risk in octogenarians remains unclear. METHODS: Two-hundred and six consecutive octogenarians (130 men, 83.3 ± 3.4 years) underwent stent implantation (104 bare metal stents [BMSs] and 102 DESs) and 38.0 ± 13.2 months of follow-up. RESULTS: Significantly more DES patients received DAT. The incidence of bleeding events was similar in the DES and BMS groups for 1 year (total: 10.8% vs 5.8%, P = 0.19; major: 4.9% vs 2.9%, P = 0.70). However, after 2 years, significantly more bleeding events occurred in the DES group than the BMS group (total: 2 years, 21.6% vs 9.6%, P = 0.02; 3 years, 29.4% vs 11.5%, P = 0.001; 4 years, 31.4% vs 15.4%, P = 0.007; major: 2 years, 12.7% vs 3.8%, P = 0.04; 3 years, 18.6% vs 5.8%, P = 0.005; 4 years, 19.6% vs 6.7%, P = 0.006). Overall, significantly more total bleeding events (31.4% vs 15.4%, P = 0.007) and major bleeding events (19.2% vs 6.7%, P = 0.006) were observed in the DES group than in the BMS group. The adjusted hazard ratios and 95% confidence intervals (CI) were as follows: total bleeding events, 2.203 (95% CI: 1.065-4.556; P = 0.033); major bleeding events, 4.324 (1.506-12.414; P = 0.007). CONCLUSIONS: DAT was associated with an increased risk of bleeding events in octogenarians after 2 years. DAT discontinuation should be considered for octogenarians 1-year post-DES implantation.

In vitro thermal effects on embryonic cells of endangered hawksbill turtle Eretmochelys imbricata.

The hawksbill turtle is an ectotherm, whose sex is determined by temperature during embryonic development. This study aimed to determine whether embryonic hawksbill turtle cells respond differently to temperature than mammalian cells. Embryonic hawksbill turtle cells were established in culture, and thermal effects on these cells were investigated in vitro. Cells were maintained in Dulbecco's Modified Eagle Medium supplemented with non-essential amino acids, vitamin solution, sodium pyruvate, and 10% fetal bovine serum at 33°C and cell proliferation occurred at 25-33°C. When cells were incubated at 37°C (the temperature of mammalian cell culture) for 24 h, cell growth was completely inhibited. This growth inhibition was evidently recovered by changing the incubation temperature back to 33°C. Expression of heat shock protein was found to increase with elevating culture temperature from 25 to 33°C.

Backup support of the mother-child technique: technical considerations for the size of the mother guiding catheter.

BACKGROUND: We previously proposed a technique called the "mother-child technique" to facilitate stent delivery for complex coronary lesions. This technique is applicable when the backup support of the guiding catheter is insufficient. In this study, we used an in vitro coronary artery tree model to determine the impact of the size of the mother guiding catheter on the backup support of the mother-child guiding system. METHODS: The backup support was measured for the 4-in-5, 4-in-6, 4-in-7, and 4-in-8 systems as well as for the 5-in-6, 5-in-7, and 5-in-8 systems. RESULTS: Advancement of the child catheter into the coronary artery tree model improved the backup support of the mother-child system. When a 4-Fr child catheter was advanced by 9 cm, the relative increase in the backup support was 174% in the 4-in-5 system; it was 203% in the 4-in-6, and 135% in the 4-in-7 system (P < 0.05 vs. the mother catheter alone). The relative increase with the 5-Fr child catheter was 289% in the 5-in-6 system, and 152% in the 5-in-7 system (P < 0.0001 vs. the mother catheter alone). However, the improvement in the backup support with 9-cm child catheter advancement did not reach statistical significance for either the 4-in-8 (115%) or the 5-in-8 (112%) system (P = NS). CONCLUSIONS: The relative increase in the backup support of the mother-child system was inversely related to the size of the mother guiding catheter. Thus, the mother-child technique may be most useful for PCIs in which a small guiding catheter is used, such as transradial coronary interventions.

Outcomes of primary percutaneous coronary intervention for acute myocardial infarction with unprotected left main coronary artery occlusion.

BACKGROUND: We determined the in-hospital and the long-term outcomes of primary percutaneous coronary intervention (PCI) for acute left main coronary artery (LMCA) occlusion. METHODS: Between 1988 and 2009, 72 patients with acute myocardial infarction (AMI) underwent primary PCI for unprotected LMCA occlusion. The short- and the long-term outcomes of primary PCI in these patients were retrospectively evaluated. RESULTS: Upon arrival, cardiogenic shock was observed in 33 (46%) patients and cardiopulmonary arrest (CPA) in 12 (17%). Twenty-three (32%) required extracorporeal life support and 64 (89%) intra-aortic balloon pumping. Although successful reperfusion was achieved in 60 (83%) patients, in-hospital death was observed in 32 (44%). Multivariate analysis revealed predictors of in-hospital death to be CPA on arrival (RR, 7.05; 95% CI: 1.28-39.0; P = 0.025). During 1.7 ± 2.9 years of follow-up, 10 of the 40 hospital survivors died. All presenting CPA on arrival died within 2 years. Although the estimated survival of the all study patients was only 26.2% at 8 years by the Kaplan-Meier methods, those without shock/CPA on arrival showed nearly flat survival curve after 4 years. CONCLUSIONS: Despite performance of primary PCI, patients with AMI due to LMCA occlusion were associated with >50% in-hospital mortality. Hemodynamic deficit upon arrival was the major determinant of their poor hospital outcomes. The hospital survivors, however, were associated with favorable long-term outcomes.

Enantioselective nitroso aldol reaction catalyzed by QuinoxP*.silver(I) complex and tin methoxide.

A catalytic enantioselective O-nitroso aldol reaction of alkenyl trichloroacetates with nitrosoarenes was achieved using the (R,R)-t-Bu-QuinoxP*.AgOAc complex as the chiral catalyst and Bu(2)Sn(OMe)(2) as the achiral cocatalyst in the presence of methanol. Optically active alpha-aminooxy ketones with up to 99% ee were regioselectively obtained in high yields from various alkenyl trichloroacetates of cyclic ketones.

Percutaneous coronary intervention using a virtual 3-Fr guiding catheter.

BACKGROUND: We have recently reported a novel percutaneous coronary intervention (PCI) system using a hydrophilic-coated sheathless guiding catheter (Virtual 3-Fr, Medikit, Tokyo, Japan), which provides us with less invasive angioplasty and a puncture site injury equivalent to a conventional 3-Fr introducer sheath. Here, we report the initial results of PCI using this novel system. METHODS: A total of 36 coronary artery lesions of 27 patients were treated by using a virtual 3-Fr PCI system. Procedural outcomes of virtual 3-Fr PCI were retrospectively evaluated. RESULTS: The mean age was 73.0 +/- 8.7 years (range, 46-84 years), and 15 were men (56%). Access sites included the radial artery in 18 patients (67%), the brachial artery in eight patients (30%), and the femoral artery in 1 patients (4%). Among 36 lesions, seven were chronic total occlusions, and a virtual 3-Fr PCI was successful in 33 lesions (92%). Among the successfully treated 33 lesions, coronary stents were deployed in 32 (97%), and intravascular ultrasound examination was performed in 19 (58%). Hemostasis was achieved immediately after PCIs in all cases. No access-site related complications including radial artery occlusion were observed. CONCLUSIONS: The performance of a virtual 3-Fr PCI system appears to be comparable to one using a regular 5-Fr guiding catheter while the puncture-site damage remains equivalent to that of a 3-Fr introducer sheath. Virtual 3-Fr PCI may have a potential to serve as a minimally invasive strategy for the treatment of coronary artery diseases.

Coronary intervention with 4-French catheters.

OBJECTIVES: We sought to determine whether 4-Fr percutaneous coronary intervention (PCI) is associated with technical difficulties that might have an unfavorable impact on procedural parameters. BACKGROUND: Four-Fr PCI is often associated with difficulties in catheter manipulation, which may lead to greater time consumption and increased dye usage when compared with PCI employing larger guiding catheters. METHODS: From July 2007 to March 2009, 62 patients underwent 4-Fr PCI. Procedural characteristics were compared between patients who underwent 4-Fr PCI in 2007 (early phase: 31 lesions in 26 patients) and those underwent in 2008 or later (later phase: 40 lesions in 36 patients). RESULTS: Ad-hoc coronary intervention (3% vs. 23%, P < 0.05) and deep-vessel intubation (46% vs. 91%, P < 0.05) were observed less frequently in the late phase than the early phase. Fluoroscopy time (8 +/- 6 min vs. 17 +/- 15 min, P < 0.05) and the amount of contrast dye used (64 +/- 33 mL vs. 90 +/- 46 mL, P < 0.05) were significantly reduced in the late phase than the early phase. No access site-related complications were observed in patients in either phase. CONCLUSIONS: The performance of 4-Fr PCI requires a certain learning curve, following which a reduction in fluoroscopy time and use of contrast dye may be achieved. This improvement in procedural parameters and the low incidence of access site-related complications might allow 4-Fr PCI to serve as a minimally invasive approach for the treatment of coronary artery diseases.

Genetic analysis of young adult patients with aortic disease not fulfilling the diagnostic criteria for Marfan syndrome.

BACKGROUND: Although the existence of the young patients with aortic disease not fulfilling the diagnostic criteria for Marfan syndrome (MFS) has been known, the etiology of their disease has not yet been elucidated. The purpose of the present study was to elucidate the genetic and clinical features of the young patients with aortic disease not having MFS. METHODS AND RESULTS: Eighty young adult patients with aortic disease were examined. They were divided into a definite MFS (n=51) and a non-definite MFS group (n=29) according to the Ghent nosology. Clinical and genetic characteristics were compared between the 2 groups. Among 29 non-definite MFS probands, 1 (3%) FBN1, 2 (7%) TGFBR1, and 3 (10%) TGFBR2 mutations were found, and 4 ACTA2 mutations were found in the 23 probands examined without FBN1, TGFBR1, or TGFBR2 mutations. In total, more than 10 out of 29 (34%) probands in the non-definite MFS group were associated with genetic mutations. Skeletal involvement was less frequent in the non-definite than in the definite MFS group (7% vs 82%, P<0.01). CONCLUSIONS: In the probands with aortic diseases in young who cannot be diagnosed with MFS, mutations other than FBN1 mutations accounted for at least one-third of all causes of aortic disease.

Wortmannin, a specific inhibitor of phosphatidylinositol-3-kinase, induces accumulation of DNA double-strand breaks.

Wortmannin, a fungal metabolite, is a specific inhibitor of the phosphatidylinositol 3-kinase (PI3K) family, which includes double-stranded DNA dependent protein kinase (DNA-PK) and ataxia telangiectasia mutated kinase (ATM). We investigated the effects of wortmannin on DNA damage in DNA-PK-deficient cells obtained from severe combined immunodeficient mice (SCID cells). Survival of wortmannin-treated cells decreased in a concentration-dependent manner. After treatment with 50 µM wortmannin, survival decreased to 60% of that of untreated cells. We observed that treatment with 20 and 50 µM wortmannin induced DNA damage equivalent to that by 0.37 and 0.69 Gy, respectively, of γ-ray radiation. The accumulation of DNA double-strand breaks (DSBs) in wortmannin-treated SCID cells was assessed using pulsed-field gel electrophoresis. The maximal accumulation was observed 4 h after treatment. Moreover, the presence of DSBs was confirmed by the ability of nuclear extracts from γ-ray-irradiated SCID cells to produce in vitro phosphorylation of histone H2AX. These results suggest that wortmannin induces cellular toxicity by accumulation of spontaneous DSBs through inhibition of ATM.

Transradial coronary intervention using a novel 5-Fr sheathless guiding catheter.

We have recently developed a 5-Fr sheathless guiding catheter system (Virtual 3-Fr, Medikit, Japan), composed of a hydrophilic catheter and a central dilator. This combination of catheter and dilator enables us to introduce the catheter into the artery without the need of an introducer sheath. Because the outer diameter of this 5-Fr sheathless guiding catheter system is approximately 2-Fr sizes smaller than a conventional guiding catheter system, this system provides us with less invasive angioplasty and a puncture site injury equivalent to a conventional 3-Fr introducer sheath. To demonstrate the potential of this 5-Fr sheathless guiding system, we report two cases of chronic total occlusion successfully treated by the transradial approach. This novel 5-Fr sheathless system may become a viable alternative to conventional guiding catheters, and provide a favorable impact upon vascular access complications and patient morbidity.

Single injection of a sustained-release prostacyclin analog improves pulmonary hypertension in rats.

RATIONALE: Although prostacyclin is recognized as a therapeutic breakthrough for pulmonary hypertension, it needs continuous infusion because of its short action. Therefore, we developed a new drug delivery system for prostacyclin. We prepared ONO-1301MS, a novel sustained-release prostacyclin analog polymerized with poly(d,l-lactic-co-glycolic acid) (PLGA) microspheres. OBJECTIVES: We examined whether ONO-1301MS attenuates monocrotaline (MCT)-induced pulmonary hypertension in rats, and attempted to elucidate the underlying mechanisms responsible for the beneficial effects of ONO-1301MS. METHODS: After MCT injection, rats were randomized to receive a single subcutaneous injection of 100 mg/kg ONO-1301MS or vehicle. MEASUREMENTS AND MAIN RESULTS: We prepared ONO-1301MS, which was polymerized with PLGA to release ONO-1301 for 3 weeks. A single administration of ONO-1301MS achieved sustained elevation of its circulating level and plasma cyclic adenosine 3',5'-monophosphate level for 3 weeks, and attenuated an increase in a metabolite of thromboxane A(2) level. Rats had developed pulmonary hypertension 3 weeks after MCT injection; however, treatment with ONO-1301MS significantly attenuated the increases in right ventricular systolic pressure and right ventricular weight to body weight ratio. ONO-1301MS significantly inhibited hypertrophy of pulmonary arteries. Phosphorylation of extracellular signal-regulated protein kinase (ERK) in the lung was significantly increased in the control group, whereas this increase was markedly attenuated by treatment. CONCLUSIONS: We developed a new drug delivery system for prostacyclin using PLGA and ONO-1301. A single injection of ONO-1301MS resulted in sustained activity for 3 weeks, and attenuated pulmonary hypertension, partly through its antiproliferative effect on vascular smooth muscle cells via inhibition of ERK phosphorylation.

Usefulness of fibrinogen/fibrin degradation product to predict poor one-year outcome of medically treated patients with acute type B aortic dissection.

Previous studies have indicated that medical therapy provides excellent outcomes for patients with uncomplicated Stanford type B acute aortic dissection. However, affected aortas are often compromised by aneurysmal dilatation and rupture, resulting in poor outcomes. The purpose of this study was to determine predictors of aortic events in patients with Stanford type B acute aortic dissection receiving conservative medical therapy. The study group consisted of 78 consecutive patients with Stanford type B acute aortic dissection who were admitted to the hospital within 48 hours of onset. These patients were treated medically and followed up for 1 year. Aortic events were defined as rupture, recurrent dissection, aortic expansion with diameter>or=60 mm, rapid aortic expansion at a rate of >or=10 mm/yr, and the development of visceral or limb ischemia. Predictors of these events were determined using multivariate analyses. During 1-year follow-up, aortic events were observed in 13 (17%) patients, including aortic rupture in 3 (4%), aortic diameter>or=60 mm in 4 (5%), rapid expansion of the aorta in 3 (4%), and the development of visceral or limb ischemia in 3 (4%). On multivariate analysis, fibrinogen-fibrin degradation product level>or=20 microg/ml (odds ratio 7.802, 95% confidence interval 1.405 to 43.335) on admission was the only independent predictor of aortic events at 1 year. In conclusion, careful monitoring is required for patients with medically treated Stanford type B acute aortic dissection associated with fibrinogen-fibrin degradation product level>or=20 microg/ml on admission.

Differences in atherosclerotic profiles between patients with thoracic and abdominal aortic aneurysms.

Differences in atherosclerotic profiles between patients with thoracic aortic aneurysm (TAA) and patients with abdominal aortic aneurysm (AAA) have not been studied. We retrospectively studied the clinical records of 343 consecutive patients (132 TAA and 211 AAA) who were admitted to our hospital for elective repair of aortic aneurysms between July 2001 and December 2004. Clinical variables were compared between patients with TAA and those with AAA by using a univariate analysis, and those achieving statistical significance were subsequently assessed in a multivariate analysis. The incidence of coronary artery disease (CAD) (53% vs 23%, p <0.0001), 3-vessel coronary disease (41% vs 10%, p <0.0001), male gender (86% vs 74%, p <0.01), smoker (88% vs 76%, p <0.01), chronic obstructive pulmonary disease (COPD) (30% vs 15%, p <0.01), and diabetes mellitus (39% vs 23%, p <0.01) were significantly higher in patients with AAA than in those with TAA. In contrast, the incidence of hypertension (91% vs 81%, p <0.05), saccular-type aneurysm (61% vs 7%, p <0.0001), and body mass index (24.1 +/- 3.1 vs 23.2 +/- 3.5, p <0.05) were significantly higher in patients with TAA than in those with AAA. Multivariate stepwise logistic analysis revealed that CAD (odds ratio [OR] 3.65; 95% confidence interval [CI] 2.12 to 6.42; p <0.0001), COPD (OR 2.05; 95% CI 1.11 to 3.89; p <0.05), and diabetes mellitus (OR 1.85; 95% CI 1.06 to 3.27; p <0.05) were associated with AAA, and that body mass index (OR 9.39; 95% CI 2.0 to 46.8; p <0.01), hypertension (OR 3.09; 95% CI 1.48 to 6.87; p <0.01), and cerebral infarction (OR 2.83; 95% CI 1.25 to 6.50; p <0.05) were associated with TAA. In conclusion, atherosclerotic profiles are significantly different between patients with TAA and patients with AAA. This result suggests the possibility that mechanisms underlying the development of aortic aneurysms may differ between TAA and AAA, and, from the perspective of prevention, provides further stimulus for the modification of key risk factors for atherosclerosis.

A prodigiosin analogue inactivates NADPH oxidase in macrophage cells by inhibiting assembly of p47phox and Rac.

Prodigiosins are natural red pigments that have multi-biological activities. Recently, we discovered a marine bacterial strain, which produces a red pigment. Extensive two-dimensional nuclear magnetic resonance and mass spectrometry analysis showed that the pigment is a prodigiosin analogue (PG-L-1). Here, we investigated the effect of PG-L-1 on NADPH oxidase activity in macrophage cells. PG-L-1 significantly inhibited superoxide anion (O(2)(-)) production by phorbol 12-myristate 13-acetate (PMA)-stimulated RAW 264.7 cells, a mouse macrophage cell line. The ED(50) value was estimated to be approximately 0.3 microM. PG-L-1 had no direct scavenging effect on O(2)(-) generated by hypoxanthine/xanthine oxidase system in electron spin resonance-spin trapping determinations, suggesting that this compound directly acts on the O(2)(-) production system, NADPH oxidase, in macrophage cells. We further investigated the effect of PG-L-1 on the behaviour of the cytosolic components of the NADPH oxidase, p67(phox), p47(phox), p40(phox), Rac and protein kinase C (PKC), in PMA-stimulated RAW 264.7 cells. Although PG-L-1 showed no effect on the activation of PKC, the immunoblotting analysis using specific antibodies showed that PG-L-1 strongly inhibits the association of p47(phox) and Rac in the plasma membrane of PMA-stimulated RAW 264.7 cells. These results suggest that PG-L-1 inactivates NADPH oxidase through the inhibition of the binding of p47(phox) and Rac to the membrane components of NADPH oxidase.

Percutaneous coronary intervention using a novel 4-French coronary accessor.

BACKGROUND: Percutaneous coronary intervention (PCI) using a guiding catheter with small diameters may have a favorable impact on vascular access complications and patient morbidity. Here, we report the initial results of PCI using a 4-Fr coronary accessor. METHODS: A total of 31 patients underwent 4-Fr PCI. Exclusion criteria for 4-Fr PCI were (1) lesions associated with large side branches requiring wire protection or kissing balloon technique and (2) planned use of angioplasty devices which were not compatible with 4-Fr catheter. RESULTS: A total of 36 lesions, including 4 chronic total occlusions (CTO), were treated. Access sites included radial artery in 19 patients (61%), brachial artery in 8 (26%), and femoral artery in 4 (13%). Four-Fr PCI was successful in 34 of 36 lesions (94%) in 29 of 31 patients (94%). One of the two unsuccessful patients was a case of CTO, and the other a case of tortuous right coronary artery. In both, crossover to a 6-Fr PCI was necessary. Among successfully treated 34 lesions of the 29 patients, coronary stents were deployed in 30 lesions (88%). There were no stent dislodgements or inadequate contrast opacification. No access-site related complications including radial artery occlusion were observed. CONCLUSIONS: PCI with a 4-Fr coronary accessor is a viable alternative to the use of larger guide catheters. The advent of 4-Fr stent delivery system may afford a less invasive approach for the treatment of patients with coronary artery disease.

The effect of ice crystals formations on the flesh quality of frozen horse mackerel (Trachurus japonicus).

It is known that the formation of ice crystals has a negative impact on the flesh quality of frozen meat. This study focuses on how the formation of ice crystals in the dorsal ordinary muscle affects the flesh quality of frozen horse mackerel (Trachurus japonicus). Freeze-thawed muscle specimens (F-TMS) of horse mackerel were first stored at -24C and then thawed. The K-value, expressible water, breaking strength of unfrozen muscle specimens (UMS) and F-TMS, and histological structure (light microscope [LM], scanning electron microscope [SEM] of UMS, frozen muscle specimens [FMS], and F-TMS) were investigated. K-values were higher in the F-TMS than in the UMS, and the increasing rate of K-value in F-TMS was approximately 1.4-fold higher than UMS. Similarly, the percentage of expressible water was higher in the F-TMS than in the UMS. The breaking strength of the UMS decreased with storage period, but it remained at a higher level than that of F-TMS. Numerous ice crystal formations in muscle cells of FMS and small interstices between cells of F-TMS were observed under LM. Moreover, we observed a relatively collapsed honeycomb structure of the extracellular matrix (ECM) under SEM, and found that collagen fibril networks of endomysiums became looser after thawing. Therefore, ice crystals contribute to vulnerabilities in the ECM. These results suggest that a decrease of freshness and the degradation of physical properties caused by ECM destruction may be due to the formation of ice crystals in muscle structures during freezing, leading to the deterioration of flesh quality during storage. PRACTICAL APPLICATIONS: The flesh quality deteriorated after thawing on the basis of degradation of physical properties. Moreover, it could be presumed that the K-value increase was exacerbated by ice crystals, and the decreasing speed of chemical freshness was sped up accompanied by ice storage after thawing. Thus, the formation of ice crystals, which destroyed the muscle cells and structures, may be one of the dominant factors causing the deterioration in the flesh quality.