Transcarotid vascular access for transcatheter aortic valve implantation: is choosing the left side always right?

BACKGROUND: The transcarotid (TC) vascular access for transcatheter aortic valve implantation (TAVI) has emerged as the first-choice alternative to the transfemoral access, in patients unsuitable for the latter. The use of both the left and right common carotid arteries (CCAs) for TC-TAVI has been described, but the optimal side is subject to debate. We conducted this pilot study to compare the level of vessel tortuosity and plaque burden from either the left CCA to the aortic annulus, or the right CCA to the aortic annulus, considering them as surrogates for technical and procedural complexity. METHODS: Consecutive patients who underwent TC-TAVI between 2018 and 2021 in our institution were included. Using three-dimensional reconstruction, pre-TAVI neck and chest computed tomography angiography exams were reviewed to assess the tortuosity index (TI), sum of angles metric, as well as plaque burden, between each CCA and the aortic annulus. RESULTS: We included 46 patients who underwent TC-TAVI. No significant difference regarding the mean TIs between the left and right sides (respectively 1.20 and 1.19, p = 0.82), the mean sum of angles (left side: 396°, right side: 384°, p = 0.27), and arterial plaque burden (arterial plaque found in 30% of left CCAs and 45% of right CCAs, p = 0.19) was found. CONCLUSIONS: We found no convincing data favoring the use of one particular access side over the other one. The choice of the CCA side in TC-TAVI should to be made on a case-by-case basis, in a multidisciplinary fashion, and may also depend on the operators' experience.

Facilitating Repeat Intracarotid Injections in Mouse Models by a Novel Injection Site Repair Technique.

Given recent advances in the delivery of novel antitumor therapeutics using endovascular selective intraarterial delivery methods in neuro-oncology, there is an urgent need to develop methods for intracarotid injections in mouse models, including methods to repair the carotid artery in mice after injection to allow for subsequent injections. We developed a method of intracarotid injection in a mouse model to deliver therapeutics into the internal carotid artery (ICA) with two alternative procedures. During injection, the needle is inserted into the common carotid artery (CCA) after tying a suture around the external carotid artery (ECA) and injected therapeutics are delivered into the ICA. Following injection, the common carotid artery (CCA) can be ligated, which limits the number of intracarotid injections to one. The alternative procedure described in this article includes a modification where intracarotid artery injection is followed by injection site repair of the CCA, which restores blood flow within the CCA and avoids the complication of cerebral ischemia seen in some mouse models. We also compared the delivery of bone marrow-derived human mesenchymal stem cells (BM-hMSCs) to intracranial tumors when delivered through intracarotid injection with and without injection site repair following the injection. Delivery of BM-hMSCs does not differ significantly between the methods. Our results demonstrate that injection site repair of the CCA allows for repeat injections through the same artery and does not impair the delivery and distribution of injected material, thus providing a model with greater flexibility that more closely emulates intracarotid injection in humans.

Morphology of the aortic arch branching pattern in raccoon dogs (Nyctereutes procyonoides, Gray, 1834).

BACKGROUND: Aortic arch (AA) branching patterns vary among different mammalian species. Most previous studies have focused on dogs, whereas those on raccoon dogs remain unexplored. OBJECTIVES: The objective of this study was to describe the AA branching pattern in raccoon dogs and compare their morphological features with those of other carnivores. METHODS: We prepared silicone cast specimens from a total of 36 raccoon dog carcasses via retrograde injection through the abdominal aorta. The brachiocephalic trunk (BCT) branching patterns were classified based on the relationship between the left and right common carotid arteries. The subclavian artery (SB) branching pattern was examined based on the order of the four major branches: the vertebral artery (VT), costocervical trunk (CCT), superficial cervical artery (SC), and internal thoracic artery (IT). RESULTS: In most cases (88.6%), the BCT branched off from the left common carotid artery and terminated in the right common carotid and right subclavian arteries. In the remaining cases (11.4%), the BCT formed a bicarotid trunk. The SB exhibited various branching patterns, with 26 observed types. Based on the branching order of the four major branches, we identified the main branching pattern, in which the VT branched first (98.6%), the CCT branched second (81.9%), the SC branched third (62.5%), and the IT branched fourth (52.8%). CONCLUSIONS: The AA branching pattern in raccoon dogs exhibited various branching patterns with both similarities and differences compared to other carnivores.

TRIGLYCERIDE - GLUCOSE INDEX, REMNANT CHOLESTEROL AND COMMON CAROTID ARTERY INTIMA-MEDIA THICKNESS AS AN ATHEROSCLEROTIC MARKER IN ISCHEMIC STROKE PATIENTS.

Ischemic stroke (IS) is a major global health concern, often resulting from atherosclerosis and insulin resistance (IR). The triglyceride-glucose index (TyG index), remnant cholesterol (RC), and common artery intima-media thickness (CIMT) are potential markers for assessing atherosclerosis and cardiovascular risk in IS patients. A cross-sectional study was conducted to investigate the association between TyG index, RC, CIMT, and IS in adult patients recruited from a hospital. Demographic, clinical, and laboratory data were collected, and statistical analysis was performed. The study included 50 participants with a balanced gender distribution and a mean age of 57.64 years. Laboratory characteristics showed notable values, and CIMT > 0.6 mm was associated with higher NIH Stroke Scale scores. RC exhibited significant correlations with age, CIMT, lipid profile, and TyG index. The study highlights the potential of TyG index, RC, and CIMT as atherosclerotic markers in IS patients. Favorable prognostic outcomes were observed, emphasizing the importance of early diagnosis and management to improve patient outcomes.

Balloon neck-plasty to create a wide-necked aneurysm in the elastase-induced rabbit model.

PURPOSE: The elastase-induced aneurysm (EIA) model in rabbits has been proposed for translational research; however, the adjustment of aneurysm neck size remains challenging. In this study, the technical feasibility and safety of balloon neck-plasty to create a wide-necked aneurysm in rabbit EIA model were investigated. METHODS: Male New Zealand White rabbits (N = 15) were randomly assigned to three groups: group A, EIA creation without neck-plasty; group B, neck-plasty immediately after EIA creation; group C, neck-plasty 4 weeks after EIA creation. The diameter of balloon used for neck-plasty was determined 1 mm larger than origin carotid artery diameter. All rabbits were euthanized 4 weeks after their final surgery. Aneurysm neck, height, dome-to-neck (D/N) ratio, and histologic parameters were compared among the groups. RESULTS: Aneurysm creation was technically successful in 14 out of 15 rabbits (93.3%), with one rabbit experiencing mortality due to an adverse anesthetic event during the surgery. Saccular and wide-necked aneurysms were successfully created in all rabbits. Aneurysm neck was significantly greater in groups B and C compared to group A (all P < .05). D/N ratio was significantly lower in groups B and C compared to group A (all P < .05). Additionally, tunica media thickness, vessel area, and luminal area were significantly greater in groups B and C compared to group A (all P < .05). These variables were found to be significantly greater in group B compared to group C (all P < .05). CONCLUSION: The creation of a wide-necked aneurysm using balloon neck-plasty after elastase induction in rabbits has been determined to be technically feasible and safe.

CD38 deficient mice are not protected from atherosclerosis.

CD38 is a multifunctional enzyme implicated in chemotaxis of myeloid cells and lymphocyte activation, but also expressed by resident cells such as endothelial and smooth muscle cells. CD38 is important for host defense against microbes. However, CD38's role in the pathogenesis of atherosclerosis is controversial with seemingly conflicting results reported so far. To clarify the discrepancy of current literature on the effect of CD38 ablation on atherosclerosis development, we implanted a shear stress modifier around the right carotid artery in CD38-/- and WT mice. Hypercholesterolemia was induced by human gain-of-function PCSK9 (D374Y), introduced using AAV vector (serotype 9), combined with an atherogenic diet for a total of 9 weeks. Atherosclerosis was assessed at the aortic root, aortic arch and the right carotid artery. The findings can be summarized as follows: i) CD38-/- and WT mice had a similar atherosclerotic burden in all three locations, ii) No significant differences in monocyte infiltration or macrophage content could be seen in the plaques, and iii) The amount of collagen deposition in the plaques were also similar between CD38-/- and WT mice. In conclusion, our data suggest that CD38-/- mice are neither protected against nor prone to atherosclerosis compared to WT mice.

Feasibility and Intermediate Results of Transcarotid Revascularization with a Prosthetic Conduit.

BACKGROUND: Transcarotid artery revascularization (TCAR) is a hybrid technique with excellent initial outcomes. The technical success and safety of TCAR is heavily dependent on an anatomically suitable common carotid artery (CCA). Many patients do not meet anatomic criteria and therefore are not eligible for this therapy. We sought to extend the eligibility of TCAR to patients with unfavorable CCA anatomy via the adoption of a prosthetic arterial conduit. METHODS: A single-center retrospective study of patients with critical carotid artery stenosis who underwent TCAR via a prosthetic conduit between June 2019 and October 2021 was performed. All patients in the study were considered high-risk for carotid endarterectomy based on anatomic features, such as restenosis post-carotid endarterectomy and neck radiation. Unfavorable CCA anatomy was defined as a clavicle to carotid bifurcation distance <5 cm, a CCA diameter <6 mm, and/or significant atherosclerotic disease at the intended arterial access site. The primary outcome of interest was technical success. Secondary outcomes included perioperative complications, intermediate and long-term patency, intermediate and long-term stroke and/or mortality and in-hospital length of stay. Follow-up ranged from 1 to 29 months. RESULTS: Eight patients underwent 10 TCAR procedures via a prosthetic conduit. A total of 2 procedures (20%) were performed on female patients and 8 procedures (75%) were performed on male patients. The mean age was 65 years old (standard deviation 11 years). Technical success was 100%. The 30-day ipsilateral stroke rate was 0%. The 30-day patency was 90%. There was no re-exploration for hemorrhage and 30 day mortality was 0%. CONCLUSIONS: TCAR is an excellent option for carotid artery revascularization. Unfavorable CCA anatomy has limited its applicability. TCAR via a prosthetic conduit has the potential to expand eligibility for this promising therapy.

Arterial Health Markers in Relation to Behavior and Cognitive Outcomes at School Age.

BACKGROUND: Impaired arterial health is associated with a decline in cognitive function and psychopathology in adults. We hypothesized that these associations originate in early life. We examined the associations of blood pressure, common carotid artery intima media thickness, and carotid distensibility with behavior and cognitive outcomes during adolescence. METHODS AND RESULTS: This study was embedded in the Dutch Generation R Study, a population-based prospective cohort study from early fetal life onwards. Blood pressure, carotid intima media thickness, and carotid distensibility were measured at the age of 10 years. At the age of 13 years, total, internalizing and externalizing problems and attention-deficit hyperactivity disorder symptoms were measured using the parent-reported Child Behavior Checklist (CBCL/6-18), autistic traits were assessed by the Social Responsiveness Scale, and IQ was assessed using the Wechsler Intelligence Scale for Children-Fifth Edition. A 1-SD score higher mean arterial pressure was associated with lower odds of internalizing problems (odds ratio [OR], 0.92 [95% CI, 0.85-0.99]). However, this association was nonsignificant after correction for multiple testing. Carotid intima media thickness and carotid distensibility were not associated with behavior and cognitive outcomes at 13 years old. CONCLUSIONS: From our results, we cannot conclude that the associations of blood pressure, carotid intima media thickness, and carotid distensibility at age 10 years with behavior and cognitive outcomes are present in early adolescence. Further follow-up studies are needed to identify the critical ages for arterial health in relation to behavior and cognitive outcomes at older ages.

A thyroidea ima artery variation providing collateral circulation to the mediastinum.

The thyroidea ima artery (TIA) is a highly variable arterial deviation of the blood supply to the thyroid gland with critical implications for surgical neck procedures such as tracheostomy. Though relatively common in the population at large (~ 4%), most TIA variations are related to the origin of the artery and whether it emerges from the common sites of the brachiocephalic trunk, aortic arch, and right common carotid artery, or another more unique vessel-as opposed to its dispersion pattern. TIA variants generally supply the thyroid gland, occasionally co-occurring with absent thyroid arteries. Here, we report on a unique case of a four-pronged variation of the TIA discovered during an anatomy laboratory dissection of first-year medical students. This variant originated from the brachiocephalic trunk and had three branches terminating in the thyroid gland and a fourth branch traveling into the thorax to provide accessory circulation in the mediastinum. Specifically, small arterial branches from the inferior TIA branch supplied the anterior pericardium and surrounding adipose tissue, in addition to normal pericardiacophrenic circulation. We discuss the potential embryological and clinical relevance of this unique variation and voice further support for imaging as a requirement before surgical neck procedures to prevent catastrophic bleeding in the event of a TIA variant.

Sentinel ear bleed as a presenting sign for carotid artery blowout due to aspergillus abscess in parapharyngeal space in an immunocompetent infant.

Parapharyngeal abscesses leading to complications, although rare after the advent of antibiotics, can lead to serious complications. One such complication is carotid erosion that can lead to a potentially fatal carotid artery blowout. We report a case of a previously healthy infant who presented with fever, ear bleed and progressively increasing swelling in the right side of his neck that led to airway compromise. The child required immediate securing of the airway at presentation. Imaging revealed lobulated abscess with multiple bleeding points eroding the carotid vessels, along with internal jugular venous thrombus. Surgical exploration was done and abscess debulked. Histopathology revealed aspergillus, which was treated with antifungals. He was discharged on oral warfarin after 40 days of hospital stay and remains well on follow-up. Sentinel ear bleed warrants close observation for possibility of carotid artery blowout in children with parapharyngeal abscesses.

Thyrolinguofacial trunk arising from the common carotid artery and diagnosed by computed tomography angiography.

PURPOSE: Various variations in the head and neck vasculature have been reported. The purpose of this report is to describe an extremely rare case of thyrolinguofacial trunk (TLFT) arising from the common carotid artery (CCA). METHODS: A 66-year-old woman with vertigo, dizziness, and heaviness in the head underwent computed tomography (CT) angiography of the neck and head region for evaluation of cerebrovascular diseases. RESULTS: The TLFT originated from the anterior wall of the right CCA and was divided into the superior thyroid artery and linguofacial trunk (LFT). The LFT was divided into lingual and facial arteries. In addition, we observed fusiform dilatation of the intracranial right vertebral artery, which might have caused these symptoms. CONCLUSION: The presence of a common trunk of the external carotid artery (ECA) branches increases the risk of complications such as bleeding and ischemia during treatment of the head and neck region, including chemoradiotherapy for oral bleeding and tongue cancer. Therefore, this is an area of significant interest across various medical specialties, including surgery, otolaryngology, and radiology. Understanding the diverse variations in the neck vasculature is expected to lead to a reduction in complications associated with various procedures.

Preparation of a Single-Cell Suspension from Mouse Carotid Arteries for Single-Cell Sequencing.

Carotid arteries are major blood vessels in the neck that supply blood and oxygen to the brain, but carotid stenosis occurs when carotid arteries are clogged by plaque. Revealing the cellular composition of the carotid artery at the single-cell level is essential for treating carotid atherosclerosis. However, there is no ready-to-use protocol for the preparation of single-cell suspensions from carotid arteries. To obtain a suitable protocol for the dissociation of normal carotid arteries at the single-cell level with less damage to cells, we designed a two-step digestion method by integrating the digestion process of collagenase/DNase and trypsin. Acridine orange/propidium iodide (AO/PI) dual-fluorescence counting was used to detect cell viability and concentration, and it was found that the single-cell suspension satisfied the requirements for single-cell sequencing, with the viability of cells over 85% and a high cell concentration. After single-cell data processing, a median of ~2500 transcripts per cell were detected in each carotid artery cell. Notably, a variety of cell types of the normal carotid artery, including vascular smooth muscle cells (VSMCs), fibroblasts, endothelial cells (ECs), and macrophages and dendritic cells (Mφ/DCs), were concurrently detectable. This protocol may be applied to prepare a single-cell suspension of blood vessels from other tissues with appropriate modifications.

Is low carotid bifurcation determined by vertebral level always convenient for surgical approach?

Although high-level carotid bifurcation (HCB) could lead to notable surgical difficulty, the definitive reference point for HCB is unclear. HCB is typically characterized as carotid bifurcation (CB) located higher than the level of the third cervical vertebra (C), however, a major obstacle regarding carotid artery surgical exposure is angle of the mandible (AM). The objective of this study was to investigate CB level, define HCB in relation to AM and vertebral levels, and measure the vertical distance from HCB to ipsilateral AM. Moreover, the percentage of surgically challenged CBs, misclassified as low CBs (LCB) based on vertebral level, was investigated. Patients who underwent neck computed tomography angiography were retrospectively studied. HCBs were classified into two categories: CBs above the C3 and either at or above the ipsilateral AM. Of 172 CBs (86 patients; 57 men, 29 women), CB was mostly found at C3 (44.19%), whereas AM was commonly located at C2 (51.16%). Based on vertebral level and AM, HCBs were detected in 10.47% and 20.35% of CBs, respectively. The association of HCBs determined by either C3 or AM between both sides in each individual was nonsignificant (p>0.05), but HCBs determined by C3 level were predominant in women (OR = 3.58, 95%CI = 1.31-9.80). Considering both C3 and AM, there was 8.72% of HCBs. The remaining 91.28% was classified as LCBs, including 11.63% of CBs located at both C3 and AM which were actually classified as HCBs if determined by AM. In cases of CBs above AM level, the mean vertical distance was as high as 6.56 ±2.41mm.

Trans-Distal Radial Artery Carotid Revascularization with Forearm Flow Reversal: An Alternative Option of CAS in the TCAR Era.

BACKGROUND: Transcarotid artery revascularization (TCAR) has emerged as an alternative to carotid artery stenting (CAS). TCAR demonstrated its superiority by avoiding femoral artery puncture and establishing proximal protection without crossing the stenotic lesion. In the TCAR era, we focused on the possibility of a trans-distal radial approach (DRA). A balloon-guide catheter was navigated via DRA to establish proximal protection before lesion crossing. The forearm subcutaneous vein was used as the flow-reversal circuit. METHODS: Six internal carotid artery stenosis patients underwent CAS using "the forearm flow reversal technique." Every procedure was performed under continuous flow reversal from the common carotid artery to the forearm cephalic vein. RESULTS: Successful revascularization was achieved without ischemic or access-site complications. The distal radial artery was patent at discharge in all cases. CONCLUSIONS: Trans-distal radial CAS with forearm flow reversal is a feasible and less invasive technical option.

Reconstructive interpolation for pulse wave estimation to improve local PWV measurement of carotid artery.

Ultrasonic transit time (TT)-based local pulse wave velocity (PWV) measurement is defined as the distance between two beam positions on a segment of common carotid artery (CCA) divided by the TT in the pulse wave propagation. However, the arterial wall motions (AWMs) estimated from ultrasonic radio frequency (RF) signals with a limited number of frames using the motion tracking are typically discrete. In this work, we develop a method involving motion tracking combined with reconstructive interpolation (MTRI) to reduce the quantification errors in the estimated PWs, and thereby improve the accuracy of the TT-based local PWV measurement for CCA. For each beam position, normalized cross-correlation functions (NCCFs) between the reference (the first frame) and comparison (the remaining frames) RF signals are calculated. Thereafter, the reconstructive interpolation is performed in the neighborhood of the NCCFs' peak to identify the interpolation-deduced peak locations, which are more exact than the original ones. According to which, the improved AWMs are obtained to calculate their TT along a segment of the CCA. Finally, the local PWV is measured by applying a linear regression fit to the time-distance result. In ultrasound simulations based on the pulse wave propagation models of young, middle-aged, and elderly groups, the MTRI method with different numbers of interpolated samples was used to estimate AWMs and local PWVs. Normalized root mean squared errors (NRMSEs) between the estimated and preset values of the AWMs and local PWVs were calculated and compared with ones without interpolation. The means of the NRMSEs for the AWMs and local PWVs based on the MTRI method with one interpolated sample decrease from 1.14% to 0.60% and 7.48% to 4.61%, respectively. Moreover, Bland-Altman analysis and coefficient of variation were used to validate the performance of the MTRI method based on the measured local PWVs of 30 healthy subjects. In conclusion, the reconstructive interpolation for the pulse wave estimation improves the accuracy and repeatability of the carotid local PWV measurement.

Cascaded Plane Wave Ultrasound for Blood Velocity Vector Imaging in the Carotid Artery.

Cascaded dual-polarity waves (CDWs) imaging increases the signal-to-noise ratio (SNR) by transmitting trains of pulses with different polarity order, which are combined via decoding afterward. This potentially enables velocity vector imaging (VVI) in more challenging SNR conditions. However, the motion of blood in between the trains will influence the decoding process. In this work, the use of CDW for blood VVI is evaluated for the first time. Dual-angle, plane wave (PW) ultrasound, CDW-coded, and noncoded conventional PW (cPW), was acquired using a 7.8 MHz linear array at a pulse repetition frequency (PRF) of 8 kHz. CDW-channel data were decoded prior to beamforming and cross correlation-based compound speckle tracking for VVI. Simulations of single scatterer motion show a high dependence of amplitude gain on the velocity magnitude and direction for CDW-coded transmissions. Both simulations and experiments of parabolic flow show increased SNRs for CDW imaging. As a result, CDW outperforms cPW VVI in low SNR conditions, based on both bias and standard deviation (SD). Quantitative linear regression and qualitative analyses of simulated realistic carotid artery blood flow show a similar performance of CDW and cPW for high SNR (14 dB) conditions. However, reducing the SNR to 6 dB, results in a root-mean-squared error 2.7× larger for cPW versus CDW, and an R2 of 0.4 versus 0.9. Initial in vivo evaluation of a healthy carotid artery shows increased SNR and more reliable velocity estimates for CDW versus cPW. In conclusion, this work demonstrates that CDW imaging facilitates improved VVI of deeper located carotid arteries.

Patients with a large ossification of the posterior longitudinal ligament have a higher incidence of arteriosclerosis in the carotid artery.

PURPOSE: This study aimed to investigate the impact of the severity of cervical ossification of the posterior longitudinal ligament (OPLL) on the incidence of arteriosclerosis in the carotid artery. METHODS: Patients with OPLL-induced cervical myelopathy were prospectively enrolled. The study involved analyzing patient characteristics, blood samples, computed tomography scans of the spine, and intima-media thickness (IMT) measurements of the common carotid artery. Patients were divided into two groups based on the size of the cervical OPLL to compare demographic data, comorbidities, and the presence of thickening of the carotid intima-media (max IMT ≥ 1.1 mm). RESULTS: The study included 96 patients (mean age: 63.5 years; mean body mass index: 26.9 kg/m2; 71.8% male; 35.4% with diabetes mellitus). The mean maximum anteroposterior (AP) diameter of the OPLL was 4.9 mm, with a mean occupancy ratio of 43%. The mean maximum IMT was 1.23 mm. Arteriosclerosis of the carotid artery was diagnosed in 62.5% of the patients. On comparing the two groups based on OPLL size, the group with larger OPLL (≥ 5 mm) had a higher BMI and a greater prevalence of carotid intima-media thickening. This significant difference in the prevalence of carotid intima-media thickening persisted even after adjusting for patient backgrounds using propensity score matching. CONCLUSIONS: Patients with a larger cervical OPLL showed a higher frequency of intima-media thickening in the carotid artery.

Association of cumulative low-density lipoprotein cholesterol exposure with vascular function.

BACKGROUND: The relationship between cumulative low-density lipoprotein cholesterol (LDL-C) exposure and progression of atherosclerosis remains uncertain. OBJECTIVE: The aim of this study was to determine the relationship between cumulative LDL-C level and flow-mediated vasodilation (FMD), nitroglycerine-induced vasodilation (NID) and the presence of plaque in the common carotid artery (CCA). METHODS: This was a cross-sectional study. We measured FMD in 8208 subjects, NID in 1822 subjects, and CCA plaque in 591 subjects who were not taking lipid-lowering drugs. The subjects were divided into four groups based on cumulative LDL-C exposure: <4000 mg·year/dL, 4000-4999 mg·year/dL, 5000-5999 mg·year/dL, and ≥6000 mg·year/dL. RESULTS: The odds ratio of the lower quartile of FMD in the cholesterol-year-score <4000 mg·year/dL group was significantly higher than the odds ratios in the other groups. The odds ratio of the lower quartile of NID in the <4000 mg·year/dL group was significantly higher than the odds ratios in the 5000-5999 mg·year/dL and ≥6000 mg·year/dL groups. The odds ratio of the prevalence of CCA plaque in the <4000 mg·year/dL group was significantly higher than that in the ≥6000 mg·year/dL group. CONCLUSIONS: Endothelial dysfunction occurred from cumulative LDL-C exposure of 4000 mg·year/dL, vascular smooth muscle dysfunction occurred from cumulative LDL-C exposure of 5000 mg·year/dL, and prevalence of CCA plaque occurred from cumulative LDL-C exposure of 6000 mg·year/dL. CLINICAL TRIAL REGISTRY INFORMATION: http://www.umin.ac.jp (UMIN000012950, UMIN000012951, and UMIN000012952, UMIN000003409).