Association of insulin-like growth factor-1 receptor gene polymorphisms with left ventricular mass and geometry in essential hypertension.

Stimulation of insulin-like growth factor (IGF)-1 receptor by IGF-1 and insulin strongly induces cardiomyocyte hypertrophy. In this study, we assessed the hypothesis that genetic variations of the IGF-1 receptor may be linked to the diversity of left ventricular (LV) structure in hypertensive patients. Genotypes in 12 single nucleotide polymorphisms (SNPs) of the IGF-1 receptor gene identified by direct sequencing were determined in 795 Japanese patients with essential hypertension. In echocardiographic examinations, LV mass index (LVMI) and relative wall thickness (RWT) were measured. Among 12 SNPs, promoter -328C>T and intron-13 275124A>C polymorphisms were significantly associated with LV hypertrophy (LVMI> or =125 g m(-2)) and concentric change (RWT> or =0.44), respectively. In allele frequencies, the C allele of -328C>T was related to LV hypertrophy, and the A allele of 275124A>C was related to LV concentric change. In fact, LVMI and prevalence of LV hypertrophy increased in CC genotype of -328C>T. RWT and prevalence of LV concentric change increased in AA genotype of 275124A>C. A multiple logistic regression analysis revealed that the presence of CC genotype of -328C>T or AA genotype of 275124A>C was an independent determinant for LV hypertrophy or concentric change, respectively. Furthermore, the combination of CC of -328C>T and AA of 275124A>C genotypes was significantly associated with abnormal LV geometry, especially concentric hypertrophy. Our findings show that two SNPs of the IGF-1 receptor gene are related to LV hypertrophy in patients with essential hypertension, suggesting that the genetic variation of the IGF-1 receptor may be involved in the diversity of LV structure in hypertensives.

[Safety measures of extracorporeal circulation by heart surgeons and perfusionists].

In the past, heart surgeons often set up the extracorporeal circulation (ECC) system, primed the circuit, and operated the ECC in Japan. As works of perfusionists recently became specialized, young Japanese heart surgeons seldom receive education on ECC, and rarely operate ECC. ECC accidents are rare, but once it occurs, even a well experienced perfusionist often becomes too upset to think of the next action, while surgeons at the operative table have little knowledge of the ECC system. Reconsideration of ECC education for heart surgeons is still rare. As a medical team, tragedies such as death and life-threatening complication due to an ECC accident are to be prevented at all costs. At an on-site training session for ECC troubles at the 59th annual meeting of Japanese Association for Thoracic Surgery, the basic ECC operations, recovering procedures after an accident, and the use of safety devices were taught to 30 teams of young heart surgeons and perfusionists as a measure to ensure safety of ECC. A questionnaire survey was conducted at the end and satisfactory results were obtained.

Association of single nucleotide polymorphisms in endothelin family genes with the progression of atherosclerosis in patients with essential hypertension.

Endothelin-1 (ET-1) is a potent vasoconstrictive peptide and its activity is mediated by the receptors ET type A (EDNRA) and ET type B (EDNRB). Although ET-1 is thought to play an important role in the development of atherosclerosis, it remains unclear whether polymorphisms of ET-1 family genes, including the ET-1 gene (EDN1), EDNRA, EDNRB and the genes for endothelin converting enzymes 1 and 2 (ECE1 and ECE2), are associated with the progression of atherosclerosis. We investigated the relationship between 11 single nucleotide polymorphisms (SNPs) of ET-1 family genes (including three in EDN1, one in EDNRA, two in EDNRB, four in ECE1 and one in ECE2) and atherosclerotic changes assessed using pulse wave velocity (PWV) and carotid ultrasonography in 630 patients with essential hypertension (EHT). In male subjects, we found significant differences in brachial-ankle PWV (baPWV) in additive and recessive models in EDNRB-rs5351 after Bonferroni correction. Also in male subjects, there were significant differences in mean intima-media thickness (IMT) in additive and recessive models in EDNRA-rs5333 after Bonferroni correction. We found no significant correlation between any SNPs in the ET family genes and baPWV, IMT and Plaque score (PS) in female subjects. Furthermore, after multiple logistic regression analysis, only EDNRB-rs5351 indicated as an independent risk of atherosclerosis in male hypertensive subjects. Of the endothelin-related genes, EDNRB-rs5351 was the most susceptible SNP associated with atherosclerosis in male hypertensives, and the genetic background may be involved in the progression of atherosclerosis in EHT patients.

Induction of osteoclast-like cells derived from the synovial lavage fluids of patients with temporomandibular joint disorders.

OBJECTIVE: Although biochemical studies have examined the synovial fluid (SF) of patients with temporomandibular joint (TMJ) disorders (TMDs), the details of the molecular mechanism of bone destruction and remodeling remain unknown. In this study, we induced and characterized osteoclast-like cells from the SF of patients with TMD and investigated the participation of these cells in the pathogenesis of TMD. METHODS: We collected SF cells from patients with TMD after a pumping procedure, cultured osteoclast-like cells, and examined their characteristics, including osteoclast markers and bone resorption activities. In addition, we obtained fibroblastic cells from the SF of TMD patients by continuous sub-culturing. Using these fibroblastic cells, we examined fibroblast markers using immunocytochemical staining and analyzed the receptor activator of nuclear-factor-kappaB ligand (RANKL) mRNA levels. Detection of soluble form of RANKL (sRANKL) in the SF was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Osteoclast-like cells were induced from the SF cells of patients with TMD by adding recombinant human (rh) macrophage colony stimulating factor (M-CSF) and either 1,25-dihydroxy vitamin D3 [1,25(OH)2D3] or prostaglandin E2 (PGE2). These multinucleated giant cells were positive for tartrate-resistant acid phosphatase (TRAP) and had the ability to absorb bone. The fibroblastic cells from the SF of TMD patients were positive for fibroblast markers and RANKL mRNA was up-regulated. Detection of sRANKL in SF of patient group was significantly higher than control group. CONCLUSION: The results suggest that the joint-infiltrating SF cells from TMD patients play important roles in the pathogenesis of these disorders, which is characterized by progressive bone destruction or remodeling.

Optimal low-energy laser irradiation causes temporal G2/M arrest on rat calvarial osteoblasts.

Low-energy laser irradiation (LELI) accelerates wound healing and is thought to accelerate bone formation. However, the mechanism of laser healing is not clear. To clarify the biological mechanism of LELI healing, we investigated the effects of LELI on rat osteoblasts in vitro. Osteoblastic cells from 3-day-old Wistar rat calvaria were irradiated using a low-energy gallium-aluminum-arsenide (Ga-Al-As) diode laser. Bone formation, osteoblast differentiation, and cell proliferation were evaluated by von Kossa staining, reverse-transcription polymerase chain reaction, alkaline phosphatase (ALP) staining, 5-bromo-2'-deoxyuridine (BrdU) uptake, and fluorescence-activated cell sorter (FACS) analysis. At 21 days after LELI, the greatest bone formation was observed with irradiation energy of 3.75 J/cm2 and the first week after seeding. LELI (3.75 J/cm2) induced an increased number of cells at day 3. LELI-stimulated differentiation in osteoblastic cells was demonstrated by the increases of Runx2 expression and ALP-positive colonies. By contrast, at 1 day after laser irradiation, the number of cells in the irradiation group was significantly lower than that in the control group. BrdU uptake indicated lower proliferation 12 and 24 hours after irradiation compared with the control. Furthermore, FACS data demonstrated a higher proportion of cells in the G2/M phase of the cell cycle 12 hours after irradiation compared with the control. G2/M arrest was confirmed by the appearance of G2/M arrest marker 14-3-3-sigma or phospho-p53. These results demonstrate that LELI induces not only acceleration of bone formation but also initial G2/M arrest, which may cause wound healing like tissue repair.

Functional reorganization of the spinal pain pathways in developmental and pathological conditions.

Following inflammation, a subpopulation of Abeta afferents that terminates preferentially in deeper laminae have been shown to extend their axons to the superficial dorsal horn, particularly substantia gelatinosa (SG). Similarly, SG neurons in immature spinal cord receive mainly Abeta afferent inputs. To clarify whether the reorganized sensory pathway in the inflamed rats has a functional similarity with that in the developmental state, we compared synaptic inputs from primary afferents using in vitro and in vivo patch-damp recordings from SG neurons. SG neurons in the mature state had monosynaptic inputs from Adelta and C afferents, while only a few neurons received inputs from Abeta afferents. Following inflammation, the Abeta afferents extended their axons to SG and established functional monosynaptic transmission. Meanwhile, SG neurons in the immature state received preferentially Abeta as well as Adelta afferent inputs, and the majority of Abeta afferent inputs were monosynaptic. These observations support the idea that the sprouting of the large afferent fibres observed in inflamed rats is, at least in part, a regeneration process. However, the process, maybe distinct at some point from the process during development, therefore, produces pathological pain. Though the idea that the regeneration mimics the developmental process has been widely accepted, other possibilities cannot be excluded.

Diagnostic value of carotid intima-media thickness and plaque score for predicting target organ damage in patients with essential hypertension.

Carotid intima-media thickness (IMT) assessed by ultrasonography is regarded as an early predictor of general arteriosclerosis in patients with essential hypertension. However, the methods of measuring IMT have not been globally standardized, and it remains unclear whether conventional measurement of IMT represents the prevalence of hypertensive target organ damage. In this study, we verified the association between several commonly used carotid ultrasonographical parameters and the severity of hypertensive target organ damage (retinal arteriosclerosis, microalbuminuria, left ventricular hypertrophy (LVH)). Carotid ultrasonography, echocardiography, urinalysis, and funduscopy were performed in 184 patients (64 +/- 12 years, 96 males and 88 females) with various stages of essential hypertension. Carotid arteriosclerosis was assessed using four methodologically different methods: conventional-IMT, maximum-IMT (Max-IMT), Mean-IMT, and Plaque Score (the sum of all plaque thicknesses). Age and all carotid ultrasonographical parameters were significantly associated with albuminuria, retinal arteriosclerosis, and left ventricular mass index. High-sensitivity CRP was significantly correlated with retinopathy and LVH. Carotid parameters in patients with histories of cardiovascular events were significantly greater in those without events. Among all carotid parameters, Max-IMT showed the highest correlation coefficient of the severity of target organ damage, and showed significant association with CRP. Stepwise regression analysis revealed that Max-IMT was the independent factor for predicting target organ damage. Max-IMT is suggested to be the most reliable and simplest parameter for predicting hypertensive target organ damage including microangiopathy in patients with essential hypertension.

Association between first trimester absence of fetal nasal bone on ultrasound and Down syndrome.

OBJECTIVES: To evaluate the association between absence of nasal bone on ultrasound and Down syndrome in fetuses at 11-14 weeks of pregnancy. METHODS: One hundred and ninety-four consecutive fetuses from singleton pregnancies undergoing chorionic villi sampling (CVS) were evaluated by transabdominal ultrasound. A sagittal view of the fetal face was obtained and the absence or presence of nasal bone was recorded and correlated with the fetal karyotype. RESULTS: A successful view of the fetal profile was possible in 183 of 194 (94%) fetuses. The nasal bone was absent in three of five fetuses with Down syndrome, and in one of 175 (0.6%) chromosomally normal fetuses. CONCLUSION: Absence of nasal bone by first trimester ultrasound was significantly associated with Down syndrome. When a proper view of the fetal face was obtained, the nasal bone was visible in more than 99% of karyotypically normal fetuses.

Efficiency of an air filter at the drainage site in a closed circuit with a centrifugal blood pump: an in vitro study.

In a closed circuit with a centrifugal blood pump, one of the serious obstacles to clinical application is sucking of air bubbles into the drainage circuit. The goal of this study was to investigate the efficiency of an air filter at the drainage site. We used whole bovine blood and the experimental circuit consisted of a drainage circuit, two air filters, a centrifugal blood pump, a membrane oxygenator, a return circuit, and a reservoir. Air was injected into the drainage circuit with a roller pump, and the number and size of air bubbles were measured. The air filter at the drainage site could remove the air bubbles (>40 microm) by itself, but adding a vacuum removed more bubbles (>40 microm) than without vacuum. Our results suggest that an air filter at the drainage site could effectively remove air bubbles, and that adding the filter in a closed circuit with a centrifugal blood pump would be safer.

Intraoperative assessment of coronary artery bypass graft: transit-time flowmetry versus angiography.

BACKGROUND: Transit-time flowmetry has been used to assess graft status intraoperatively. This study examines the validity of this method by comparing its results with the findings of simultaneously performed graft angiography. METHODS: The left internal thoracic artery (LITA) anastomosed to the left anterior descending artery (LAD) was assessed intraoperatively with both transit-time flowmetry and graft angiography in 30 patients. The patients were stratified into two groups based on intraoperative angiographic findings. In 18 patients (group A), the LITA and the LAD were well filled with contrast medium and the anastomosis was widely patent. In the other 12 patients (group B), spastic LITA or LAD was observed. Postoperative angiography was also performed before discharge from the hospital. RESULTS: The mean graft flow was 44.0 +/- 25.4 mL/min in group A and 23.4 +/- 10.0 mL/min in group B (p = 0.0129). Diastolic-dominant flow pattern was observed in both groups, and the ratio of peak diastolic flow to peak systolic flow and the percent diastolic time-flow integral were not statistically different between the groups. The pulsatility index was almost the same between the two groups and was acceptable in both. Postoperative angiography revealed that all grafts were patent without spasm or anastomotic stenosis. CONCLUSIONS: LITA graft status is satisfactory when high graft flow with diastolic dominance is obtained. When there is vasospasm but no anastomotic problems, decreased graft flow with an acceptable pulsatility index and diastolic augmentation is observed.

A new endo aortic occlusion balloon for limited access cardiac surgery: development and clinical evaluation.

In recent years, minimally invasive cardiac surgery (MICS), or limited access cardiac surgery, has been presented as a promising operative procedure. We developed a new balloon device that is inserted directly into the ascending aorta to stop the heart during limited access cardiac surgery. The balloon has a three lumen structure: balloon lumen port, cardioplegia/vent lumen port, and aortic root lumen port. This direct EAC balloon catheter, designed to be inserted directly into the ascending aorta, is different from the Heartport system. The Heartport EAC balloon catheter is inserted into the aorta via an artery in the lower limb, making lower limb arterial disease a key concern. Our Direct Endo Aortic Clamp (EAC) balloon overcomes this problem. The device was clinically used in seven cardiac cases. All patients were discharged within 5 postoperative days, confirming the utility of the device.

Closed circuit cardiopulmonary bypass with centrifugal pump for open-heart surgery: new trial for air removal.

The purpose of this study is to examine the efficiency of venous air removal with a new cardiopulmonary bypass (CPB) circuit design for conventional open-heart surgeries. A main concern with a closed circuit for open-heart surgeries is air entrainment into the venous line. A venous filter was placed proximal to the centrifugal pump. The circuit proximal to the centrifugal pump was divided into two lines; one line was attached to the venous reservoir outlet. By clamping the line to the reservoir, this circuit becomes closed. Negative pressure was applied to the purge line connected to the venous reservoir for venous air removal. Micro bubbles were measured at two locations, both distal to the venous and arterial filters. When the injection rate reached 100 ml/min, with the air-injection over 30 s, micro bubbles greater than 40 micro were observed at the outlet of venous filter. However, there was no micro bubble greater than 40 micro detected at the outlet of arterial filter. Although micro bubbles greater than 40 micro were not detected at the outlet of the arterial filter up to the injection rate of 300 ml/min, when the injection rate reached 400 ml/min, micro bubbles greater than 50 microm were detected distal to the arterial filter. From this examination, we determined that air entrained in the venous line up to approximately 300 ml/min is automatically removed by this method with the pressure-balanced condition. This pressure balance means that resistance of venous return, gravity siphon, negative pressure by centrifugal pump, and negative pressure applied to the air-purge line of the filter are balanced; that is, the venous return is sufficient, and the venous reservoir volume is kept stable. From this study we determined that this circuit design efficiently removes the entrained air in the venous line.

Vacuum assisted cardiopulmonary bypass in minimally invasive cardiac surgery: its feasibility and effects on hemolysis.

The present study describes a cardiopulmonary bypass (CPB) technique that incorporates vacuum assisted venous drainage and arterial return using a centrifugal pump in minimally invasive cardiac surgery (MICS). The technique was performed on 40 patients scheduled to undergo MICS. The proposed CPB technique enables a good operative field to be obtained even through a limited incision through the use of peripheral cannulation using small cannulae. Vacuum pressure was applied to the venous reservoir (-43 +/- 14 mm Hg) to maintain adequate CPB flow (>2.4 L x min-1 x M-2). The effects of CPB on hemolysis were subsequently compared between patients who underwent the proposed procedure (MICS group; n = 6) and a control group who underwent coronary arterial bypass grafting (CABG group; n = 6) with conventional CPB. Plasma free hemoglobin (FHb) increased and plasma haptoglobin (Hp) decreased during CPB in both groups, with no significant difference between the groups. By the next day, FHb had returned to pre-CPB levels whereas Hp remained lower in both groups. Again, these values did not differ significantly between groups. Thus, we conclude that the proposed CPB technique is useful in MICS with acceptable effects on hemolysis.

Development of a completely closed circuit using an air filter in a drainage circuit for minimally invasive cardiac surgery.

The completely closed circuit system is the future direction of cardiopulmonary bypass because of its compactness and superior biocompatibility. The most serious obstacle for clinical application is the sucking of air bubbles into the drainage circuit. The purpose of this study was to remove the air bubbles from the drainage circuit. Infusing 50 ml/min of air bubbles into the drainage circuit of the usual closed circuit, and infusing 50, 100, and 150 ml/min of air into the drainage circuit of a newly developed closed circuit (drainage circuit using an air filter), the number and size of air bubbles were observed at the outlet of the arterial filter. In the usual closed circuit, many air bubbles of over 40 microm were detected within 5 s at a blood flow of 4 L/min because the centrifugal pump decreased the size of the bubbles, which then passed through the oxygenator and arterial filter. Air bubbles of over 40 micro were not detected in the newly developed closed circuit within 5 min at a blood flow of 4 L/min. The removal of air mixed into the completely closed circuit was possible with a drainage circuit using an air filter that was developed. The clinical use of the completely closed circuit for minimally invasive cardiac surgery (MICS) became possible based on this development.

Minimally invasive port-access coronary artery bypass grafting.

The Port-Access endovascular cardiopulmonary bypass system (Heartport, Inc., Redwood City, CA, USA), a recent technological innovation in minimally invasive cardiac surgery, was conducted successfully in coronary artery bypass grafting on a 69-year-old woman. The left internal thoracic artery was harvested through a limited left anterior thoracotomy and anastomosed to the left anterior descending coronary artery on a protected and arrested heart. Intraoperative coronary angiography confirmed good graft patency. The patient was discharged from the hospital in good condition 7 days after the operation. This was the first successful minimally invasive Port-Access coronary artery bypass grafting in Japan.