Cepharanthine induces the proliferation of human dermal papilla cells and stimulates vascular endothelial growth factor expression through increased intracellular calcium mobilization and hypoxia-inducible factor activation.

BACKGROUND: Cepharanthine (CEP), a compound extracted from the vine Stephania cephalantha, is commonly prescribed to treat alopecia areata; however, the scientific evidence for its efficacy is limited. AIM: To investigate the effect of CEP and its structural analogues on human hair growth in vitro. METHODS: The effects of CEP and three of its structural analogues on the proliferation of human dermal papilla cells (hDPCs) and human outer root sheath cells (hORSCs) were investigated. Their effects on vascular endothelial growth factor (VEGF) expression were also assessed by real-time PCR. Activation of pathways leading to VEGF expression, such as intracellular Ca2+ mobilization and hypoxia-inducible factor (HIF) expression, was also characterized. RESULTS: CEP and two of its structural analogues significantly stimulated the growth of hDPCs but not hORSCs. Moreover, CEP and all three structural analogues significantly induced the expression of VEGF in hDPCs. CEP increased the intracellular Ca2+ concentration in hDPCs. CEP also increased the expression of HIF-1α and HIF-2α and induced the expression of HIF-responsive genes in hDPCs, even under normoxia. CONCLUSIONS: These results suggest that CEP and its structural analogues have the potential to restore hair growth by promoting the proliferation of hDPCs and increasing their expression of VEGF.

Cellular locations of proteinases and association with vesicles in Porphyromonas gingivalis.

We found that locations of arginine-specific gingipain (RGP) in the cellular fractions in the crude extract, envelope, vesicles, and culture supernatants were 48%, 16%, 17%, and 31%, respectively, and the corresponding values of lysine-specific gingipain (KGP) were 47%, 10%, 7%, and 36%, respectively. Although the molecular mass of RGP in the culture supernatant had been determined as 43 kDa, and that of KGP had been as 48 kDa, molecular masses of both proteinases solubilized from the vesicles were estimated to be over 1,500 kDa, since they eluted in the void volume of the column in the gel filtration on Sephacryl S-300. There was no reduction of molecular size by the following treatment with SDS, high-concentration NaCl, or urea. Interestingly, the occurrence of the macromolecular forms could not observed in other enzymes tested such as monopeptidyl, dipeptidyl, and tripeptidyl peptidases, as well as alkaline phosphatase. Therefore, occurrence of the macromolecular forms may be restricted to the proteinases. When the vesicle and culture supernatants containing free RGP and KGP were mixed and incubated, neither RGP nor KGP seemed to bind to vesicles. RGP bound to the vesicle was found to be more stable to heat treatment than the free form, suggesting that association of RGP with the vesicle caused heat stability of this enzyme.

Hemoglobin binding activity and hemoglobin-binding protein of Prevotella nigrescens.

Prevotella nigrescens, lacking siderophores was found to bind to the hemoproteins. The binding was observed also in the envelope which was prepared by sonication of the cell. The binding occurred in the pH-dependent manner; the binding was observed below neutral pHs of the incubation mixtures but only slightly observed in the neutral and alkaline pHs. Furthermore, hemoglobin bound to the envelope was dissociated at high pHs buffers. Maximum amounts of hemoglobin bound to 1 mg envelope was 51.2 mug. Kd for the reaction at pH 5.0 was 2.1 x 10¹° M (210 pM). From the dot blot assay, hemoglobin could bind to a protein solubilized from the envelope by a detergent, referred to as hemoglobin-binding protein (HbBP), then it was purified by the sequential procedures of ion exchange chromatography, affinity chromatography and isoelectric focusing. Molecular weight and isoelectric point of the HbBP were 46 kDa and 6.1, respectively.

Analysis of amide bond formation with an alpha-hydroxy-beta-amino acid derivative, 3-amino-2-hydroxy-4-phenylbutanoic acid, as an acyl component: byproduction of homobislactone.

In the synthesis of peptidomimetics containing alpha-hydroxy-beta-amino acid, the coupling of this N(beta)-protected beta-amino acid with amine components was generally performed without the protection of its alpha-hydroxyl group. However, the formation of dipeptides in low yield was often observed when sterically hindered amine components were used. Boc-Apns-OH [Apns: (2S,3S)-3-amino-2-hydroxy-4-phenylbutanoic acid, allophenylnorstatine] (6), which is one of such beta-amino acid derivatives, is intensively employed as a core structure in the development of HIV-1 protease inhibitors. There have been no precise studies, to date, that have examined amide bond formation with alpha-hydroxy-beta-amino acid derivatives as an acyl component. To determine the cause of this low-yield reaction, we studied the amide bond formation focusing on the activation step of N(beta)-protected alpha-hydroxy-beta-amino acid by using a model coupling reaction between 6 and H-Dmt-OR [Dmt: (R)-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid] (7). A significant amount of homobislactone 9 was formed through the activation of the carboxyl group of 6 to the benzotriazole-type active esters such as OBt and OAt. In addition, this homobislactone formation was markedly increased in the presence of a catalytic amount of a base, which exhibited good correlation with the low yield of the amide bond formation, suggesting that homobislactone formation is one major reason for the low yield of the amide bond formation. Moreover, homobislactones were also formed in other derivatives of the N(beta)-protected alpha-hydroxy-beta-amino acid, suggesting a common feature of this type of amino acids. The use of a strong activation method like EDC--HOAt without base addition enhanced amide bond formation, although a small amount of homobislactone may be formed during the coupling reaction.

[Surgical treatment for thoracoabdominal and suprarenal abdominal aortic aneurysm].

We experienced 4 cases of thoracoabdominal and suprarenal abdominal aortic aneurysm in the last 6 months. There were 3 males and 1 female ranging in age from 43 to 66 years, 2 with an etiology of aortitis syndrome, 1 with non-specific inflammation and 1 with atherosclerosis. In all cases the modified Hardy procedure was employed. A 18 to 22mm X 9 to 11mm bifurcation, double velour woven dacron graft was sutured from the side of the lower descending thoracic aorta to the side of both common iliac arteries. Grafts of the same kind as above 6 to 7mm in size were attached from the main graft to major visceral arteries by end-to-end or end-to-side anastomosis. A permanent aortic-excluding clamp was attached to the normal aorta just proximal to the aneurysm. Another clamp, or double ligature with silk, was placed on the lower abdominal aorta or both common iliac arteries distal to the aneurysm. No renal and spinal preservation of any kind was employed, but renal failure and paraplegia were not recognized in any of the cases. One patient died 2 days and another 21 days after the operation due to uncontrollable bleeding and multiple organ failure. The other two are doing well.

[Several factors influencing the male sexual function in patients with atherosclerotic disease].

Patients with arteriosclerosis obliterans (ASO, n = 58), abdominal aortic aneurysm (AAA, n = 27) and the combined disease (n = 4) were the subjects of this study. All patients were evaluated by retrograde aortography and questioned personally as to their present male sexual function. The penile and ankle systolic pressure index (PSPI and ASPI) of patients with ASO were measured using a Doppler device, but in the patients with AAA, only the PSPI was taken. Several factors relating to male sexual function, such as, age, PSPI, ASPI, sites of the arterial occlusion or dilatation, classification of the ischemic symptoms according to Fontaine's category, numbers of pulsations of the femoral artery, were statistically analysed. All of the patients with both diseases, 66 percent of those with ASO and 52 percent of those with AAA were impotent. In the patients with AAA, the PSPI of the potent group was not significantly higher than that of the impotent group; however, in patients with ASO, the PSPI of the former group was significantly higher than that of the latter group. In the patients with ASO, a significant correlation between the PSPI and the ASPI (r = 0.49, p less than 0.01) was found. When a PSPI value (PSPI = 0,70) was chosen as a demarcation between the impotent group and the potent group, 84 percent of the patients with ASO were correctly classified. However, when the ASPI (ASPI = 0.45) was used, only 69 percent were properly classified.(ABSTRACT TRUNCATED AT 250 WORDS)

[Surgical treatment of abdominal aortic aneurysm in the elderly].

Because of the increasing longevity of man, more elderly patients of abdominal aortic aneurysm are being considered for surgical treatment. It is the purpose of this report to provide the data based on analysis of 23 patients, 70 years of age or older, compared with 26 patients, under 70 years of age, seen at our institution during the last 8-year period so that a rational decision can be made regarding abdominal aortic aneurysm operation in the elderly. Operation in the elderly group (n = 17) and in the younger group (n = 25) was associated with 5.9% and 4.0% operative mortality, respectively. Calculated actuarial survival at 5 years was 69.2% for operated elderly, 0% for non-operated elderly and 57.8% for operated younger groups. Rupture of abdominal aortic aneurysm was a high cause of death (40%) in the non-operated elderly group. Considering operative mortality, long-term survival and natural history of unoperated abdominal aortic aneurysms, our conclusion is that regardless of age of patients and size of aneurysms, operation should be recommended as a general rule for the elderly as for the younger patients.