The Effect of Clinical Frailty on Wound Healing in Patients With Chronic Limb-Threatening Ischemia.

PURPOSE: Clinical frailty increases the risk of adverse outcomes in older people. Patients with Chronic limb-threatening ischemia (CLTI) also had several clinical frailties. The aim of this study was to investigate the correlation between clinical frailty scales (CFS) at discharge and wound healing rate in patients with tissue loss. METHODS: A total of 510 limbs in 431 CLTI patients who were undergone endovascular treatment (EVT) from January 2013 to November 2018 were enrolled in this study. Patients were categorized into 4-groups based on the CFS stages: CFS 1 to 4 (well), CFS 5 (mild), CFS 6 (moderate) and CFS ≥7 (severe). And the change in patient's activities during hospitalize was classified into 3 groups based on CFS; improve, stable, worse. Primary endpoint was correlation between CFS at discharge and wound healing rate. Secondary endpoint was relationship between the changes in patient's activities and wound healing rate. RESULTS: A total of 365 limbs were obtained complete wound healing during this study period. Patient distribution into the 4 CFS groups was as follows: 13.3% (well), 21.8% (mild), 25.3% (moderate) and 39.6% (severe). Wound healing rate in severe CFS group was significantly lower than that in other CFS groups (p<0.0001). Wound healing rate in the patients who achieved improvement of activity was significantly higher than that in the other groups (p=0.008). CONCLUSIONS: CFS might be useful for risk stratification in patients with tissue loss. And improvement of activity during hospitalization might lead to increase the wound healing rate. CLINICAL IMPACT: Although the association between clinical frailty and prognosis outcome of Chronic limb-threatening ischemia (CLTI) has been reported, the effect of clinical frailty on wound healing remains unclear. Clinical frailty scale is independently associated with wound healing and might be useful for risk stratification in patients with tissue loss. Improvement of activity during hospitalization might lead to increase the wound healing rate.

Impact of Peak Systolic Velocity Ratio after Drug-Coated Balloon for Femoropopliteal Disease: Three-Month Serial Observation Vessel Echo Study.

AIM: No flow-limiting dissection after drug-coated balloon (DCB) treatment for femoropopliteal (FP) lesions is considered as one of the endpoints, but it has not investigated the difference between each vessel dissection. This study aimed to clarify whether there is a difference between no dissection and type C dissection without flow-limiting dissection for 3 months by peak systolic velocity ratio (PSVR) based on duplex ultrasonography. METHODS: Between February 2020 and April 2021, 44 consecutive de novo FP diseases that underwent endovascular therapy (EVT) with DCB were enrolled in this study. 65.9% of the patients had intermittent claudication, and mean lesion lengths were 194±107 mm. The chronic total occlusion was 38.6%. After DCB treatment, vessel dissection pattern was categorized by angiography. The minimum lumen area (MLA) identified by intravascular ultrasound was serially evaluated with PSVRs at 1 day, 1 month, and 3 months after EVT. RESULT: All lesions were treated with DCB without provisional stents. The vessel dissection pattern after DCB treatment showed that types D, E, and F were not observed, 9% were no dissection, 27% were type A, 32% were type B, and 32% were type C. In all cases, the PSVR values of MLA site were less than 2.6 at 3 months, and there were no significant differences between no dissection and type C dissection. CONCLUSION: Up to dissection pattern "C" is considered acceptable as one of the endpoints to determine the need for provisional stenting after DCB treatment.

Clinical Effects of Planned Endovascular Therapy for Critical Limb Ischemia Patients with Tissue Loss.

AIM: The aim of this study was to investigate the clinical effect of planned endovascular therapy (EVT) for critical limb ischemia (CLI) patients with tissue loss. Although several rounds of EVT for CLI patients are required for complete wound healing, time required for complete wound healing depends on the wound severity. We hypothesized that planned EVT might reduce the time to wound healing. METHODS: A total of 89 limbs of 76 CLI patients with tissue loss, who had undergone more than at least two EVTs were included in this study. From January 2013 through December 2015 (Conventional-EVT-group, 52 limbs), indication of target lesion revascularization (TLR) was decided based on decreased skin perfusion pressure (SPP) values or delayed wound healing. From January 2016 through October 2016 (Planned-EVT-group, 37 limbs), TLR were done every two months regardless of the SPP values until complete wound healing was obtained. Time to wound healing and complete wound healing rates were compared between the two groups. RESULTS: No significant differences existed in baseline patients and lesion characteristics between the two groups. There was no significant difference in total EVT numbers between the two groups (2.0; interquartile range, 2.0-3.0 versus 2.0; interquartile range, 2.0-3.0; P=0.9). Although complete wound healing rate was similar in both groups (71.2% versus 73.0%, p=1.0), time to wound healing was significantly shorter (95 days versus 143 days, p=0.025) in the Planned-EVT-group than in the Conventional-EVT-group. CONCLUSIONS: Planned-EVT is a useful strategy to shorten the time to wound healing for CLI patients with tissue loss.

Early development of acute kidney injury is an independent predictor of in-hospital mortality in patients with acute myocardial infarction.

BACKGROUND: Acute kidney injury (AKI) often occurs in patients with acute myocardial infarction (AMI), and is associated with adverse outcomes. However, it remains unclear how timing of AKI affects it. This study assessed impact of timing of AKI on prognosis after AMI. METHODS: This study consisted of 760 patients with AMI who were admitted within 48h after symptom onset. AKI was diagnosed as increase in creatinine ≥0.3mg/dl or ≥50% within any 48h after admission. Patients were classified into 3 groups according to the occurrence and timing of AKI: no-AKI, early-AKI (within 48h after admission) and late-AKI (>48h). Early-AKI was classified into transient early-AKI, defined as creatinine returning to the level below the criteria of AKI, and persistent early-AKI. RESULTS: Early-AKI occurred in 64 patients (9%) and late-AKI in 32 patients (4%). Patients with early-AKI had significantly higher mortality (35%) than those with late-AKI (7%, p<0.001) and no-AKI (3%, p<0.001). Multivariate analysis showed early-AKI was an independent predictor of in-hospital mortality (OR: 3.38, 95% CI: 1.30-8.76, p=0.013), but late-AKI was not. Among patients with early-AKI, mortality was significantly higher even if AKI was transient (23%, p<0.001). Patients with persistent early-AKI had the highest mortality (66%, p<0.001). CONCLUSIONS: Early-AKI was associated with worse outcome. Even if renal function once returned to baseline level, patients with early-AKI tended to be at high risk of mortality.

Effect of Intensive Statin Therapy on Coronary High-Intensity Plaques Detected by Noncontrast T1-Weighted Imaging: The AQUAMARINE Pilot Study.

BACKGROUND: Coronary high-intensity plaques detected by noncontrast T1-weighted imaging may represent plaque instability. High-intensity plaques can be quantitatively assessed by a plaque-to-myocardium signal-intensity ratio (PMR). OBJECTIVES: This pilot, hypothesis-generating study sought to investigate whether intensive statin therapy would lower PMR. METHODS: Prospective serial noncontrast T1-weighted magnetic resonance imaging and computed tomography angiography were performed in 48 patients with coronary artery disease at baseline and after 12 months of intensive pitavastatin treatment with a target low-density lipoprotein cholesterol level <80 mg/dl. The control group consisted of coronary artery disease patients not treated with statins that were matched by propensity scoring (n = 48). The primary endpoint was the 12-month change in PMR. Changes in computed tomography angiography parameters and high-sensitivity C-reactive protein levels were analyzed. RESULTS: In the statin group, 12 months of statin therapy significantly improved low-density lipoprotein cholesterol levels (125 to 70 mg/dl; p < 0.001), PMR (1.38 to 1.11, an 18.9% reduction; p < 0.001), low-attenuation plaque volume, and the percentage of total atheroma volume on computed tomography. In the control group, the PMR increased significantly (from 1.22 to 1.49, a 19.2% increase; p < 0.001). Changes in PMR were correlated with changes in low-density lipoprotein cholesterol (r = 0.533; p < 0.001), high-sensitivity C-reactive protein (r = 0.347; p < 0.001), percentage of atheroma volume (r = 0.477; p < 0.001), and percentage of low-attenuation plaque volume (r = 0.416; p < 0.001). CONCLUSIONS: Statin treatment significantly reduced the PMR of high-intensity plaques. Noncontrast T1-weighted magnetic resonance imaging could become a useful technique for repeated quantitative assessment of plaque composition. (Attempts at Plaque Vulnerability Quantification with Magnetic Resonance Imaging Using Noncontrast T1-weighted Technique [AQUAMARINE]; UMIN000003567).

Age-related decrease of meiotic cohesins in human oocytes.

Aneuploidy in fetal chromosomes is one of the causes of pregnancy loss and of congenital birth defects. It is known that the frequency of oocyte aneuploidy increases with the human maternal age. Recent data have highlighted the contribution of cohesin complexes in the correct segregation of meiotic chromosomes. In mammalian oocytes, cohesion is established during the fetal stages and meiosis-specific cohesin subunits are not replenished after birth, raising the possibility that the long meiotic arrest of oocytes facilitates a deterioration of cohesion that leads to age-related increases in aneuploidy. We here examined the cohesin levels in dictyate oocytes from different age groups of humans and mice by immunofluorescence analyses of ovarian sections. The meiosis-specific cohesin subunits, REC8 and SMC1B, were found to be decreased in women aged 40 and over compared with those aged around 20 years (P<0.01). Age-related decreases in meiotic cohesins were also evident in mice. Interestingly, SMC1A, the mitotic counterpart of SMC1B, was substantially detectable in human oocytes, but little expressed in mice. Further, the amount of mitotic cohesins of mice slightly increased with age. These results suggest that, mitotic and meiotic cohesins may operate in a coordinated way to maintain cohesions over a sustained period in humans and that age-related decreases in meiotic cohesin subunits impair sister chromatid cohesion leading to increased segregation errors.

Admission hyperglycemia is an independent predictor of acute kidney injury in patients with acute myocardial infarction.

BACKGROUND: Acute kidney injury (AKI) and acute hyperglycemia are associated with unfavorable outcomes. The impact of acute hyperglycemia on the development of AKI after acute myocardial infarction (AMI), however, remains unclear. This study was undertaken to assess the relationship between admission glucose and incidence of AKI after AMI. METHODS AND RESULTS: This study consisted of 760 patients with AMI admitted to the National Cerebral and Cardiovascular Center within 48h after symptom onset. Blood sample was obtained on admission and repeated sampling was done at least every 1 or 2 days during the first week. AKI was diagnosed as increase in serum creatinine ≥0.3mg/dl or ≥50% within any 48h. Ninety-six patients (13%) had AKI during hospitalization for AMI, and these patients had higher in-hospital mortality than those without AKI (25% vs. 3%, P<0.001). Patients with AKI had higher plasma glucose (PG) on admission than those without (222±105mg/dl vs. 166±69mg/dl, P<0.001). The incidence of AKI increased as admission PG rose: 7% with PG <120mg/dl; 9% with PG 120-160mg/dl; 11% with PG 160-200mg/dl; and 28% with PG >200mg/dl (P<0.01). On multivariate analysis admission PG was an independent predictor of AKI (odds ratio, 1.10; 95% confidence interval: 1.03-1.18, P=0.02). CONCLUSIONS: Admission hyperglycemia might have contributed to the development of AKI in patients with AMI.

Persistence of orally administered lactobacillus strains in the gut of infant mice.

The present study tested the persistence of orally administered bacteria in the gut of suckling mice. We used three bacterial strains: one strain of Lactobacillus johnsonii (designated strain Ms1) that was previously isolated from the mouse stomach, and two strains of L. plantarum, (strain No. 14 and JCM 1149(T)). We detected L. johnsonii Ms1, but neither strain of L. plantarum, in the gut 7 days after administration when the organisms were administered on days 0, 1, 3 or 7 of neonatal life. None of the strains was detected in the gut 7 days after the administration on days 14 or 28 of neonatal life. L. johnsonii Ms1 and L. plantarum JCM 1149(T) exhibited similar levels of in vitro association with gut tissues, with both strains showing association that was significantly higher than that of L. plantarum No. 14. In a separate experiment, the number of total bacteria and lactobacilli in the gut, as estimated by real-time quantitative PCR, was significantly higher in 14- and 21-day-old mice than in 0- and 7-day-old mice. In addition, the number of total bacteria was higher in 21-day-old mice than in 14-day-old mice, and the number of lactobacilli was higher in 7-day-old mice than in 0-day-old mice. These results suggest that gut persistence of administered bacteria in infant mice is species- or strain-specific and is affected by the development of indigenous microbiota. In addition, gut persistence of administered bacteria may not always depend on the tissue association capacity.

2,4-Dinitrofluorobenzene-induced contact hypersensitivity response in NC/Nga mice fed fructo-oligosaccharide.

Strategies to manipulate gut microbiota in infancy have been considered to prevent the development of allergic diseases later in life. We previously demonstrated that maternal dietary supplementation with fructo-oligosaccharide (FOS) during pregnancy and lactation modulated the composition of gut microbiota and diminished the severity of spontaneously developing atopic dermatitis-like skin lesions in the offspring of NC/Nga mice. The present study tested whether dietary FOS affects contact hypersensitivity (CHS), another model for allergic skin disease, in NC/Nga mice. In experiment 1, 5-wk-old female NC/Nga mice were fed diets either with or without FOS supplementation for 3 wk and then received 2,4-dinitrofluorobenzene (DNFB) on the ear auricle 5 times at 7-d intervals. FOS supplementation reduced CHS response as demonstrated by ear swelling. Quantitative RT-PCR analysis showed that mRNA levels for interleukin (IL)-10, IL-12p40, and IL-17 in the lesional ear skin were significantly lower in mice fed FOS. In experiment 2, female NC/Nga mice were fed diets either with or without FOS during pregnancy and lactation. After weaning, offspring were fed the diets supplemented with or without FOS. Three weeks after weaning, offspring received DNFB on the ear auricle 4 times at 7-d intervals. Although FOS supplementation after weaning reduced ear swelling, maternal FOS consumption was ineffective in offspring. The present data suggest that dietary FOS reduces CHS while maternal FOS consumption is ineffective in offspring of DNFB-treated NC/Nga mice.

Administration of antibiotics during infancy promoted the development of atopic dermatitis-like skin lesions in NC/Nga mice.

This study aimed to determine whether oral antibiotic administration during infancy is associated with the spontaneous development of atopic dermatitis-like skin lesions by modulating intestinal microbiota. Female NC/Nga mice at 3 weeks of age were orally administered kanamycin or polymyxin B. Clinical symptoms, scratching behavior, and serum antibody levels were evaluated. Changes in intestinal microbiota were determined by culture-independent analysis and cultural analysis. The kanamycin-treated mice showed higher clinical scores and scratching frequency than the control mice. IgE levels were significantly higher in the kanamycin-treated mice than in the control mice. Transient changes in intestinal microbiota were observed under kanamycin treatment. Polymyxin B treatment failed to affect scratching behavior. These results suggest that oral administration of kanamycin during infancy promoted the development of atopic dermatitis-like skin lesions in NC/Nga mice and was associated with a transient change in intestinal microbiota.

Comparison of gut microbiota and allergic reactions in BALB/c mice fed different cultivars of rice.

Our preliminary clinical trial showed that consumption of cooked rice of a Japanese common cultivar Yukihikari improved atopic dermatitis associated with a suspected rice allergy, although the underlying mechanisms remain unclear. We hypothesised that the ameliorating effect of Yukihikari on atopic dermatitis is associated with the gut microbiota. BALB/c mice were fed a synthetic diet supplemented with uncooked and polished white rice powder prepared from one of four different cultivars: Yukihikari, rice A (common rice), rice B (brewery rice) and rice C (waxy rice). Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments showed that the composition of faecal microbiota was different between mice fed Yukihikari and those fed rice A. Analysis of the 16S rRNA clone library and species-specific real-time PCR showed that the abundance of Akkermansia muciniphila, a mucin degrader, tended to be lower in mice fed Yukihikari. The incidence of allergic diarrhoea induced by oral administration of ovalbumin in systemically immunised mice was lower in mice fed Yukihikari, albeit with no difference in serum antibodies specific to ovalbumin. In a separate experiment, serum antibody levels specific to orally administered ovalbumin were lower in mice fed Yukihikari. Additionally, the translocation of horseradish peroxidase in isolated segments of ileum and colon tended to be lower in mice fed Yukihikari, suggesting a reduction in gut permeability in mice fed Yukihikari. These data indicate that changes in the gut microbiota of mice fed Yukihikari could be advantageous in the prevention of food allergy.

Maternal consumption of fructo-oligosaccharide diminishes the severity of skin inflammation in offspring of NC/Nga mice.

Strategies to manipulate the gut microbiota in infancy have been considered to prevent the development of allergic diseases later in life. We aimed to elucidate the effects of maternal dietary supplementation with a prebiotic oligosaccharide on gut microbiota and spontaneously developing atopic dermatitis-like skin lesions in the offspring of NC/Nga mice. Female NC/Nga mice were fed diets either with or without fructo-oligosaccharide supplementation during pregnancy and lactation. After weaning, offspring were fed the diets supplemented with or without fructo-oligosaccharide for 11 weeks in an air-uncontrolled conventional room. Changes in gut microbiota were assessed by denaturing gradient gel electrophoresis of the PCR-amplified 16S rRNA gene. Skin lesions were evaluated by a clinical score and scratching behaviour. Serum antibody levels were measured by ELISA, and expression levels of cytokines and chemokines in lesional tissue were evaluated by quantitative RT-PCR. Maternal supplementation with fructo-oligosaccharide modulated the gut microbiota in sucklings. Although maternal supplementation with fructo-oligosaccharide suppressed the increase in clinical skin severity score and scratching behaviour in offspring, dietary fructo-oligosaccharide after weaning was less effective. The diminution of skin lesions was accompanied by lower serum concentrations of total IgG1 and lower expression levels of TNF-alpha in the lesional tissue. These data suggest that maternal consumption of fructo-oligosaccharide diminishes the severity of atopic dermatitis-like skin lesions in the offspring of NC/Nga mice.

Role of intestinal Bifidobacterium pseudolongum in dietary fructo-oligosaccharide inhibition of 2,4-dinitrofluorobenzene-induced contact hypersensitivity in mice.

Strategies to manipulate the gut microbiota have been explored for preventing allergy development. We previously showed that dietary supplementation with fructo-oligosaccharide (FOS) reduced 2, 4-dinitrofluorobenzene (DNFB)-induced contact hypersensitivity (CHS) in BALB/c mice. Because the CHS response was negatively correlated with the number of faecal bifidobacteria, particularly Bifidobacterium pseudolongum, the present study aimed to examine whether oral administration of B. pseudolongum affects CHS response. Viable B. pseudolongum was successfully isolated from mouse faeces. Female BALB/c mice were fed a synthetic diet with or without FOS supplementation, and B. pseudolongum (2 x 10(7) cells) was administered daily throughout the experimental period. Two weeks after starting the test diets, mice received DNFB on the ear auricle twice at 7-d intervals. Conventional cultivation and molecular biological analyses based on 16S rRNA gene sequences showed that administration of FOS and B. pseudolongum resulted in higher excretion of viable bifidobacteria, mainly B. pseudolongum. Although dietary FOS reduced the CHS response as demonstrated by ear swelling, B. pseudolongum administration resulted in a reduction in the initial phase only of the CHS response. B. pseudolongum administration increased hapten-specific IgG1, while dietary FOS decreased IgG2a in sera. Administration of FOS and B. pseudolongum decreased interferon-gamma production and increased IL-10 production in cervical lymph node cells restimulated with hapten in vitro. We conclude that B. pseudolongum proliferation in the intestinal tract is partially responsible for the reduction in DNFB-induced CHS response by dietary supplementation with FOS in mice, which may be mediated by the modulation of antigen-induced cytokine production.

Response of gut microbiota to fasting and hibernation in Syrian hamsters.

Although hibernating mammals wake occasionally to eat during torpor, this period represents a state of fasting. Fasting is known to alter the gut microbiota in nonhibernating mammals; therefore, hibernation may also affect the gut microbiota. However, there are few reports of gut microbiota in hibernating mammals. The present study aimed to compare the gut microbiota in hibernating torpid Syrian hamsters with that in active counterparts by using culture-independent analyses. Hamsters were allocated to either torpid, fed active, or fasted active groups. Hibernation was successfully induced by maintaining darkness at 4 degrees C. Flow cytometry analysis of cecal bacteria showed that 96-h fasting reduced the total gut bacteria. This period of fasting also reduced the concentrations of short chain fatty acids in the cecal contents. In contrast, total bacterial numbers and concentrations of short chain fatty acids were unaffected by hibernation. Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments indicated that fasting and hibernation modulated the cecal microbiota. Analysis of 16S rRNA clone library and species-specific real-time quantitative PCR showed that the class Clostridia predominated in both active and torpid hamsters and that populations of Akkermansia muciniphila, a mucin degrader, were increased by fasting but not by hibernation. From these results, we conclude that the gut microbiota responds differently to fasting and hibernation in Syrian hamsters.

Assessing changes in composition of intestinal microbiota in neonatal BALB/c mice through cluster analysis of molecular markers.

The present study introduced a molecular biological approach to demonstrate changes in the composition of intestinal microbiota in neonatal mice. Female BALB/c mice were fed either a control diet or a diet supplemented with fructo-oligosaccharide (FOS) at 50 g/kg diet, and then mated to male mice. A cultivation-independent approach, denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified 16S rRNA gene, was performed to characterise changes in intestinal microbial populations in pups at 0, 7, 14 and 21 d old and their dams. Comparisons of DGGE profiles were performed using the Dice similarity coefficient and the unweighted pair group method with arithmetic mean (UPGMA) cluster analysis based on numbers, positions and intensities of bands. DGGE profiles differed between dams fed control and FOS-supplemented diets. Although profiles in pups on the day of birth showed a high similarity with dams, profiles in 7-d-old pups differed from dams and showed high similarity to littermates. In 14- and 21-d-old pups, profiles again showed high similarity with dams. DGGE profiles in pups were divided into two large clusters of control and FOS-supplemented diet groups in the range of 0- to 21-d-old, suggesting modulation of intestinal microbiota in infants by manipulation of microbiota in dams. The present study shows a useful technique for demonstrating changes in intestinal microbiota and provides a mouse model for modulation of intestinal microbiota in neonatal life.

Functional hypophysectomy simply by neck-strangulation in the rat: applications for ACTH and LH assays.

The classical methods for hypophysectomy require a long training-period and at the end of each experiment, it is required to check any undue remnant of the hypophyseal tissue in the sella turcica, and there must be exclusion of such datum with the remnant from the experimental group. The present method for functional hypophysectomy by neck-strangulation in the rat is very simple to perform so that even an experimenter with little experience can become easily accustomed to the technique and obtain a number of functionally hypophysectomized preparations at any time he intends, with no need to check the remnant. The cerebral blood circulation of vertebrally dislocated rats was immediately intercepted by means of neck-strangulation and artificially respired. Fifteen minutes after intravenous injection of ACTH (0.1 - 3.2 mU/rat) or LH (312.5 - 2500 ng/rat), blood was sampled from the vena cava, and sera were examined for corticosterone or testosterone by radioimmunoassay. A linear dose-response relationship was obtained within a dose range of 0.2 - 1.6 mU/rat of ACTH and 312.5 - 1250 ng/rat of LH. In the ACTH assay, parapharyngeally hypophysectomized rats were compared. It was found that the sensitivity of functionally hypophysectomized preparations was 2.72 times higher than that of the parapharyngeally hypophysectomized rats.