Kininogen-Nitric Oxide Signaling at Nearby Nonexcited Acupoints after Long-Term Stimulation.

Acupuncture treatment is based on acupoint stimulation; however, the biological basis is not understood. We stimulated one acupoint with catgut embedding for 8 weeks and then used isobaric tags for relative and absolute quantitation to screen proteins with altered expression in adjacent acupoints of Sprague Dawley rats. We found that kininogen expression was significantly upregulated in the stimulated and the nonstimulated adjacent acupoints along the same meridian. The enhanced kininogen expression was meridian dependent and was most apparent among small vessels in the subcutaneous layer. Enhanced signals of nitric oxide synthases, cGMP-dependent protein kinase, and myosin light chain were also observed at the nonstimulated adjacent acupoints along the same meridian. These findings uncover biological changes at acupoints and suggest the critical role of the kininogen-nitric oxide signaling pathway in acupoint activation.

[Surgical Management of a Severely Calcified Ascending Aorta Using Intra-Aortic Balloon Occlusion].

A severely calcified ascending aorta increases the risk of perioperative cerebral damage in cardiac surgery. Conventional aortic valve replacement using an external aortic cross clamp may be dangerous in patients with this morbidity. We used an intra-aortic balloon occlusion catheter(IABOC)to minimize risks of aortic valve replacement(AVR) in an 81-year-old man with severe aortic stenosis combined with a severely calcified aorta. IABOC was introduced to the ascending aorta via the right femoral artery by esophageal echocardiography. The precise site of the inflated balloon was confirmed by the pressure of the right radial artery and was secured by 2 tourniquets around the ascending aorta. The postoperative course was uneventful. Our technique can contribute to prevention of embolic complications in some patients with a severely calcified ascending aorta.

Mild hyperbaric oxygen activates the proliferation of epidermal basal cells in aged mice.

The proliferation of epidermal basal cells decreases with age. This study examined the effects of exposure to mild hyperbaric oxygen on the proliferative activity of epidermal basal cells in aged mouse skin. Hairless mice aged 5, 34 and 55 weeks were exposed to mild hyperbaric oxygen at 1266 hPa with 36% oxygen for 6 h/day for 1 or 2 weeks. Skin samples were then collected from the back area to evaluate epidermal thickness and the number and proliferative activity of epidermal basal cells. Exposure to mild hyperbaric oxygen had no effect on the epidermal thickness, irrespective of age, but accelerated the proliferative activity of epidermal basal cells in aged mouse skin.

A modified cryo-maze procedure via the superior transseptal approach.

The superior transseptal approach provides excellent exposure for mitral valve reconstruction. For atrial fibrillation, cryoablation is less invasive, simpler, and safer than the Cox maze operation. Combining the superior transseptal approach and cryoablation for mitral valve procedures yields a high recovery rate from atrial fibrillation, similar to that of radiofrequency ablation. Cryoablation-related complications such as thromboembolic events or requirement for a permanent pacemaker were not observed in our consecutive series. Cryoablation is also cost-effective.

The inhibitory effect of a Platycodon root extract on ultraviolet B-induced pigmentation due to a decrease in Kit expression.

The signaling of stem cell factor (SCF) through its receptor Kit is known to play an important role in regulating cutaneous melanogenesis. In the course of UVB-induced pigmentation, the expression of membrane-bound SCF by epidermal keratinocytes is upregulated at an early phase and subsequently activates neighboring melanocytes via their Kit receptors. In order to identify effective skin-lightening materials, we screened botanical extracts to determine their abilities to diminish Kit expression in melanocytes. A Platycodon root extract was consequently found to have a remarkable inhibitory activity on Kit expression. When the extract was applied to three-dimensional human skin substitutes in vitro and to human skin in vivo after UVB irradiation, their pigmentation was significantly reduced, confirming the substantial contribution of the suppression of SCF/Kit signaling to preventing or inhibiting melanin synthesis. These data demonstrate that a Platycodon root extract is a promising material for a skin-lightening product to improve pigmentation-related diseases.

A modified technique of intercostal artery implantation for thoracoabdominal aortic aneurysm using the tube of the aortic wall.

We report a modified intercostal artery implantation using the tube of the aortic wall alternative to the Crawford inclusion and Carrel Patch techniques for thoracoabdominal aortic aneurysm repair. Intercostal and lumbar arteries were isolated within a full-thickness excised aortic cuff and tailored into a tube. An 8-mm limb was sewn to the proximal graft and the aortic cuff tube. The distal tube end was sewn to an 8-mm limb and a distal limb was connected to the distal graft. Computed tomography (CT) showed no pseudoaneurysms or aneurysmal expansion but did show patent intercostal arteries at the implantation site more than 5 years later.

Graft anastomosis technique in the fragile aorta.

Here, we describe a technique for creating a secure anastomosis between a graft and the stump of the aorta, involving the fragile aortic wall between the folded flap of the graft inside the aorta and a Teflon felt strip outside.

Ultraviolet a induces endoplasmic reticulum stress response in human dermal fibroblasts.

The endoplasmic reticulum (ER) stress response is a cytoprotective mechanism against the accumulation of unfolded proteins in the ER (ER stress) that consists of three response pathways (the ATF6, IRE1 and PERK pathways) in mammals. These pathways regulate the transcription of ER-related genes through specific cis-acting elements, ERSE, UPRE and AARE, respectively. Because the mammalian ER stress response is markedly activated in professional secretory cells, its main function was thought to be to upregulate the capacity of protein folding in the ER in accordance with the increased synthesis of secretory proteins. Here, we found that ultraviolet A (UVA) irradiation induced the conversion of an ER-localized sensor pATF6α(P) to an active transcription factor pATF6α(N) in normal human dermal fibroblasts (NHDFs). UVA also induced IRE1-mediated splicing of XBP1 mRNA as well as PERK-mediated phosphorylation of an α subunit of eukaryotic initiation factor 2. Consistent with these observations, we found that UVA increased transcription from ERSE, UPRE and AARE elements. From these results, we concluded that UVA irradiation activates all branches of the mammalian ER stress response in NHDFs. This suggests that the mammalian ER stress response is activated by not only intrinsic stress but also environmental stress.

Hyperbaric oxygen improves ultraviolet B irradiation-induced melanin pigmentation and diminishes senile spot size.

BACKGROUND: The effects of exposure to hyperbaric oxygen on ultraviolet B (UVB) irradiation-induced melanin pigmentations of skins and on senile spot sizes of faces were investigated. METHODS: In the first experiment, male subjects were irradiated with UVB on their upper arms for inducing erythema and the subsequent melanin pigmentation. They were exposed to a hyperbaric environment at 1.25 atmospheres absolute (ATA) with 32% oxygen for 1 h/day, three times per week. In the second experiment, female subjects were exposed to a hyperbaric environment at 1.25 ATA with 32% oxygen for 1 h/day, two times per week. RESULTS: In the first experiment, melanin pigmentations lightened after 4 weeks of exposure to hyperbaric oxygen. In the second experiment, senile spot sizes became small after 12 weeks of exposure to hyperbaric oxygen. CONCLUSION: We concluded that exposure to hyperbaric oxygen used in this study accelerates both the fading in melanin pigmentation and the decrease in senile spot size.

Hyperbaric oxygen exposure reduces age-related decrease in oxidative capacity of the tibialis anterior muscle in mice.

The effects of exposure to hyperbaric oxygen on the oxidative capacity of the skeletal muscles in mice at different ages were investigated. We exposed 5-, 34-, 55-, and 88-week-old mice to 36% oxygen at 950 mmHg for 6 hours per day for 2 weeks. The activities of succinate dehydrogenase (SDH), which is a mitochondrial marker enzyme, of the tibialis anterior muscle in hyperbaric mice were compared with those in age-matched mice under normobaric conditions (21% oxygen at 760 mmHg). Furthermore, the SDH activities of type IIA and type IIB fibers in the muscle were determined using quantitative histochemical analysis. The SDH activity of the muscle in normobaric mice decreased with age. Similar results were observed in both type IIA and type IIB fibers in the muscle. The decrease in the SDH activity of the muscle was reduced in hyperbaric mice at 57 and 90 weeks. The decreased SDH activities of type IIA and type IIB fibers were reduced in hyperbaric mice at 90 weeks and at 57 and 90 weeks, respectively. We conclude that exposure to hyperbaric oxygen used in this study reduces the age-related decrease in the oxidative capacity of skeletal muscles.

Life cycle of human melanocytes is regulated by endothelin-1 and stem cell factor in synergy with cyclic AMP and basic fibroblast growth factor.

BACKGROUND: Although the function of human melanocytes is well characterized at cellular and molecular levels, the mechanism of the regulation of the life cycle (proliferation, differentiation, and cell death) of human melanocytes is not fully understood. OBJECTIVE: This study aims to clarify what factors are involved in regulating the life cycle of human melanocytes using serum-free culture system. METHODS: Human epidermal melanocytes were cultured in a serum-free growth medium supplemented with several kinds of growth factors, cytokines, and hormones and the effects of these factors on the life cycle of melanocytes were investigated in detail. RESULTS: Of the factors tested, endothelin-1 (ET-1) stimulated the proliferation of melanoblasts and melanocytes in the presence of cyclic AMP (cAMP)-elevating factor such as dibutyryl cAMP (DBcAMP) and of basic fibroblast growth factor (bFGF). ET-1 also stimulated the proliferation and differentiation of human melanocytes in the presence of DBcAMP. Moreover, stem cell factor (SCF) stimulated the proliferation of melanoblasts and melanocytes synergistically with ET-1. The removal of ET-1 and SCF from the culture medium greatly inhibited the proliferation of melanocytes followed by apoptotic cell death. CONCLUSION: These results suggest that the life cycle of human melanocytes is regulated by ET-1 and SCF in synergy with cAMP and bFGF.

Long-term alteration in the expression of keratins 6 and 16 in the epidermis of mice after chronic UVB exposure.

The influences of chronic UVB exposure on epidermal differentiation have been poorly studied compared to dermal photo-aging although those effects are very important in terms of photo-damage to the skin. The purpose of this study was to investigate the effects of chronic UVB exposure on keratin expression in the epidermis. The effects on murine skin of chronic exposure to weak UVB (below 1 MED) was examined by immunoblotting for keratins K10, K5, K6, and K16, by immunohistochemistry using antibodies to K6, K16, and Ki67 as well as by conventional HE staining of skin sections. Alterations of keratin expression induced by the chronic UVB exposure were distinct from those elicited by a single acute UVB exposure. The expression of keratins K6 and K16 was quite long-lasting, continuing for 7 weeks after 6 weeks of chronic UVB exposure and for 6 weeks after 9 weeks of chronic UVB exposure. In contrast, K6 and K16 expression induced by a single UVB exposure at 0.5 MED or 3 MED almost ceased within 2 weeks after that exposure. Furthermore, the expression of the constructive keratins, K5 and K10, remained almost unchanged by chronic UVB exposure. Epidermal thickness was increased significantly immediately after the 9 weeks of chronic UVB exposure; however, it had returned to normal level 6 weeks later. The alterations in keratin expression accompanied the marked disruption of the ordered ultrastructure of keratin intermediate filaments, which were observed by TEM. Thus, chronic exposure to UVB has a deep impact on the biosynthetic regulation of different keratins in the epidermis, thereby interfering with the ordered ultrastructure of keratin intermediate filaments. Those events could have relevance to the mechanism of photo-damage, such as fine wrinkles observed in chronically UV-exposed skin in addition to dermal photo-aging.

Crystal structure of the parasporin-2 Bacillus thuringiensis toxin that recognizes cancer cells.

Parasporin-2 is a protein toxin that is isolated from parasporal inclusions of the Gram-positive bacterium Bacillus thuringiensis. Although B. thuringiensis is generally known as a valuable source of insecticidal toxins, parasporin-2 is not insecticidal, but has a strong cytocidal activity in liver and colon cancer cells. The 37-kDa inactive nascent protein is proteolytically cleaved to the 30-kDa active form that loses both the N-terminal and the C-terminal segments. Accumulated cytological and biochemical observations on parasporin-2 imply that the protein is a pore-forming toxin. To confirm the hypothesis, we have determined the crystal structure of its active form at a resolution of 2.38 A. The protein is unusually elongated and mainly comprises long beta-strands aligned with its long axis. It is similar to aerolysin-type beta-pore-forming toxins, which strongly reinforce the pore-forming hypothesis. The molecule can be divided into three domains. Domain 1, comprising a small beta-sheet sandwiched by short alpha-helices, is probably the target-binding module. Two other domains are both beta-sandwiches and thought to be involved in oligomerization and pore formation. Domain 2 has a putative channel-forming beta-hairpin characteristic of aerolysin-type toxins. The surface of the protein has an extensive track of exposed side chains of serine and threonine residues. The track might orient the molecule on the cell membrane when domain 1 binds to the target until oligomerization and pore formation are initiated. The beta-hairpin has such a tight structure that it seems unlikely to reform as postulated in a recent model of pore formation developed for aerolysin-type toxins. A safety lock model is proposed as an inactivation mechanism by the N-terminal inhibitory segment.

Surgical closure of adult patent ductus arteriosus using a pursestring suture.

A 52-year-old woman with patent ductus arteriosus underwent transpulmonary surgical closure through a median sternotomy. The procedure was performed under cardiopulmonary bypass with normothermia and a beating heart, using transductal balloon occlusion and a pursestring suture around the orifice of the ductus. The use of a pursestring suture allowed minimization of the risk of balloon breakage, obviated the need for profound hypothermia and circulatory arrest, and greatly increased the technical facility of the procedure.

Production of the soluble form of KIT, s-KIT, abolishes stem cell factor-induced melanogenesis in human melanocytes.

The signaling of stem cell factor (SCF) and its receptor KIT (membrane-bound KIT; m-KIT) plays an important role in melanocyte development, survival, proliferation, and melanogenesis. It has been demonstrated in other systems that a soluble form of m-KIT released from the cell surface (s-KIT) regulates SCF signaling, although there have been no reports pertaining to the existence and the biological role of s-KIT in melanocytes. In this study, we therefore examined the involvement of s-KIT in melanogenesis. Western blotting analysis revealed that treatment with phorbol 12-myristate-13-acetate (PMA) or 4-aminophenylmercuric acetate (APMA) induced s-KIT production in cultured human melanocytes. Inhibitors of tumor necrosis factor-alpha-converting enzyme (TACE) and metalloproteinases (MMPs) muted this release of s-KIT into the media. Human recombinant s-KIT added to melanocytes inhibited SCF-induced phosphorylation of m-KIT, resulting in suppression of SCF-induced melanogenesis. Additionally, APMA-induced s-KIT production abolished SCF-induced melanogenesis as effectively as a KIT-neutralizing antibody. Concomitantly, APMA and TACE inhibitors significantly decreased and increased melanin synthesis, respectively, in an in vitro skin model. Taken together, these findings provided an insight into the elaborate mechanism of SCF/m-KIT signaling in human melanocytes and suggested that production of s-KIT contributes to the regulation of human skin pigmentation. Journal of Investigative Dermatology (2008) 128, 1763-1772; doi:10.1038/jid.2008.9; published online 31 January 2008.

High catalase production by Rhizobium radiobacter strain 2-1.

To promote the application of catalase for treating wastewater containing hydrogen peroxide, bacteria exhibiting high catalase activity were screened. A bacterium, designated strain 2-1, with high catalase activity was isolated from the wastewater of a beverage factory that uses hydrogen peroxide. Strain 2-1 was identified as Rhizobium radiobacter (formerly known as Agrobacterium tumefaciens) on the basis of both phenotypic and genotypic characterizations. Although some strains of R. radiobacter are known plant pathogens, polymerase chain reaction (PCR) analysis showed that strain 2-1 has no phytopathogenic factor. Compared with a type strain of R. radiobacter, the specific catalase activity of strain 2-1 was approximately 1000-fold. Moreover, Strain 2-1 grew faster and exhibited considerably higher catalase activity than other microorganisms that have been used for industrial catalase production. Strain 2-1 is harmless to humans and the environment and produces catalase efficiently, suggesting that strain 2-1 is a good resource for the mass production of catalase for the treatment of hydrogen peroxide-containing wastewater.

Outcomes and hemodynamics after aortic valve replacement: a comparison of stentless versus mechanical valves.

PURPOSE: Some investigators suggest that hemodynamic outcomes may be superior with the stentless aortic bioprosthesis when compared with a mechanical valve. The goal of this study was to characterize outcomes and hemodynamic data associated with each type of valve. SUBJECTS AND METHODS: Patient outcomes and echocardiographic data were compared between 25 patients with stentless valves and 59 patients with mechanical valves. RESULTS: There were no significant differences in survival and freedom from cardiovascular adverse events between two groups. The duration of anticoagulation therapy was limited to 3 months in the stentless group. There was no significant difference in preoperative and postoperative New York Heart Association (NYHA) status when comparing the two groups, and NYHA status significantly improved in both groups (P<0.05). There was no significant difference in the echocardiographic data when comparing the two groups. CONCLUSION: Aortic valve replacement using the stentless valve and the mechanical valve provided good clinical and hemodynamic outcomes. There was no significant difference in these parameters when comparing the two groups. There may be advantages in the limited required duration of anticoagulation therapy of the stentless valve , especially in elderly patients. However, longer follow-up is required before definitive conclusions regarding the benefits of the stentless valve relative to the mechanical valve can be determined.

Reinforced edge-to-edge commissural closure with mitral valvuloplasty.

A variety of techniques has been used to repair prolapse of the commissure of the mitral valve. However, the application of these techniques may be technically challenging. A simple edge-to-edge suture approximation of anterior and posterior mitral leaflets is an effective way of restoring mitral valve competence. We use this technique with "spaghetti" for reinforcement, and annuloplasty for repair of the prolapse. We have recognized it to be a simple, durable, and reproducible technique for repair of prolapse of the mitral valve commissure.