G protein-coupled receptor 30/G protein estrogen receptor-1 (GPR30/GPER-1) is a novel membrane receptor for estrogen whose mRNA is expressed at high levels in estrogen-dependent cells such as breast cancer cell lines. However, mutations in GRP30 related to diseases remain unreported. To detect unknown mutations in the GPR30 open reading frame (ORF) quickly, the experimental conditions for high-resolution melting (HRM) analysis were examined for PCR primers, Taq polymerases, saturation DNA binding dyes, Mg(2+) concentration, and normalized temperatures. Nine known SNPs and 13 artificial point mutations within the GPR30 ORF, as well as single nucleotide variants in DNA extracted from subjects with breast cancers were tested under the optimal experimental conditions. The combination of Expand High Fidelity(PLUS) and SYTO9 in the presence of 2.0mM MgCl(2) produced the best separation in melting curves of mutations in all regions of the GPR30 ORF. Under these experimental conditions, the mutations were clearly detected in both heterozygotes and homozygotes. HRM analysis of GPR30 using genomic DNA from subjects with breast cancers showed a novel single nucleotide variant, 111C>T in GPR30 and 4 known SNPs. The experimental conditions determined in this study for HRM analysis are useful for high throughput assays to detect unknown mutations within the GPR30 ORF.

Nuclear transport is mediated by transport factors, including the importin β family members. The directionality of nuclear transport is governed by the asymmetrical distribution of the small GTPase Ran. Of note, importin β-mediated import of classical nuclear localization signal (cNLS)-containing cargo is more efficient than other Ran-dependent import pathways that do not require importin α. In this study, we characterized the role of importin α in nuclear transport by examining import efficiencies of cNLS-cargo/importin α/β complexes. We first depleted digitonin-permeabilized semi-intact cells of endogenous importin α and used the cells to show that the interaction between importin α and Nup153-a component of the nuclear pore complex (NPC)-is essential for efficient import of importin β-binding domain-containing substrates but not other cargoes that directly bind to importin β. Moreover, we found that the binding of importin α to Nup153 facilitates cNLS-mediated import, and demonstrated that importin α in import complexes and cargo-free importin α prebound to Nup153 promote efficient import of cNLS-containing proteins. This is the first in vitro study showing that in conjunction with Nup153, importin α contributes to directionally biased exit of cNLS-containing cargo to the nuclear side of NPCs.

The exclusive reaction γp→K^{+}π^{-}Σ^{+} was measured for the first time using linearly polarized photons at beam energies from 1.85 to 2.96 GeV. Angular distributions in the rest frame of the K^{+}π^{-} system were fitted to extract spin-density matrix elements of the K^{*0} decay. The measured parity spin asymmetry shows that natural-parity exchange is dominant in this reaction. This result clearly indicates the need for t-channel exchange of the κ(800) scalar meson.

Atherosclerotic complications have a significant effect on mortality in patients undergoing hemodialysis (HD) therapy. However, anti-atherosclerotic and cardioprotective effects of on-line hemodiafiltration (HDF) remain to be elucidated. We prospectively compared the anti-atherosclerotic and cardioprotective effects in two randomly divided groups, i.e. on-line HDF group (n = 13) and conventional HD group (n = 9) for 1 year. Surrogate markers were brachial-ankle pulse wave velocity (baPWV), intima-media thickness (IMT) of carotid artery as an atherosclerosis marker, and cardiac functional surrogate markers included left ventricular mass index (LVMI), ejection fraction (EF), and LV diastolic capacity represented as E/A and deceleration time (DT). LVMI in on-line HDF patients showed significant regression after 1 year of treatment (131.9 ± 25.8 to 116.5 ± 24.7 g/m(2), P = 0.03), while LVMI in HD patients did not show any significant change (148.0 ± 47.1 to 142.3 ± 35.5 g/m(2)). Levels of baPWV in HD patients showed a significant increase (11.4%) from basal levels, while on-line HDF groups showed no significant increase. Furthermore, HD patients showed significant worsening of LV diastolic capacity (E/A: from 0.87 ± 0.12 to 0.79 ± 0.08, P = 0.03), while it was not shown in on-line HDF patients. Ejection fraction and IMT did not show any significant change in both groups. Serum albumin, C-reactive protein, β2 microglobulin, blood pressure, and anti-hypertensive drug use did not change in both groups. On-line HDF showed a significant improvement in LVMI and prevented a significant worsening of baPWV or LV diastolic capacity compared with patients on conventional HD therapy.

The transcription factor signal transducer and activator of transcription 3 (STAT3) has two important phosphorylation sites, Tyr705 and Ser727, for its activation. Ser727 phosphorylation has been considered to be a secondary event after Tyr705 phosphorylation. In this study, the role and regulation of Ser727 phosphorylation in STAT3 in melanocytic cells were examined. STAT3 was phosphorylated on Ser727 in the absence of Tyr705 phosphorylation in melanocytes. 12-O-tetradecanoylphorbol-13-acetate-induced increase in cell survival activity and nuclear translocation of STAT3 was associated with Ser727 phosphorylation. Ser727 was constitutively phosphorylated in all melanoma cell lines examined irrespective of Tyr705 phosphorylation. The possible involvement of Ser727 phosphorylation in STAT3 in cell survival activity and nuclear translocation of STAT3 in melanocytes was demonstrated also in melanoma cells. The constitutive Ser727 phosphorylation in melanoma cells was partially mediated by the B-Raf-MEK-ERK1/2 pathway. Immunohistochemical studies on specimens of primary lesions of acral lentiginous melanoma revealed that Ser727 phosphorylation precedes Tyr705 phosphorylation in the early stages of melanoma progression. Our results indicate that Ser727 phosphorylation on STAT3 is not necessarily a secondary event after Tyr705 phosphorylation and suggest that it has a role in the regulation of cell survival activity and nuclear translocation of STAT3 in melanocytic cells.Journal of Investigative Dermatology advance online publication, 15 March 2012; doi:10.1038/jid.2012.45.

Low nitrogen (N) availability such as that found in both dry land and tropical regions limits plant growth and development. The relationship between the level of abscisic acid (ABA) in a plant and its growth under low-N conditions was investigated. The level of ABA in cucumber (Cucumis sativus) plants under low-N conditions was significantly higher at 10 and 20d after transplantation compared with that under sufficient-N conditions. Chlorophyll was preserved in the aerial parts of cucumber plants grown under low-N conditions in the presence of ABA, while there was no significant difference between control plants and ABA-applied plants under sufficient-N conditions. ABA suppressed the reduction of chlorophyll biosynthesis under low-N conditions but not under sufficient-N conditions. On the other hand, ABA decreased the expression of the chlorophyll degradation gene in older cucumber plants grown under both conditions. In addition, transcript and protein levels of a gene encoding a chlorophyll a/b binding protein were positively correlated with ABA concentration under low-N conditions. The chloroplasts in control plants were round, and the stack of thylakoid membranes was reduced compared with that of plants treated with ABA 10(-5)M. These results strongly suggest that ABA is accumulated in cucumber plants grown under low-N conditions and that accumulated ABA promotes chlorophyll biosynthesis and inhibits its degradation in those plants.

PURPOSE: To clarify the impact of surgical site infection (SSI) after colorectal surgery on the length of hospital stay and medical expenditure in Japan. METHODS: This was a multi-center, retrospective-matched case-control study. RESULTS: The total number of patients enrolled was 334 (167 case/control pairs). The average hospital stay after surgery was prolonged by 17.8 days (95% CI 11.9-23.5) and the average medical cost after surgery was increased by $5,938 (95% CI 3,610-8,367) in the SSI group versus the non-SSI group. Hospital charges comprised the largest among all cost categories and accounted for 53% of the additional cost. The hospital stay and medical costs both increased proportionately to the depth of the SSI, from 4.4 days and $608 for superficial incisional SSI, to 39.2 days and $14,448 for organ/space SSI. SSI caused by MRSA prolonged the hospital stay by 19.3 days and incurred an additional cost of $7,015. CONCLUSIONS: SSI clearly prolonged the hospital stay and increased medical costs. The numerical values revealed by this study reinforce the medical-economic importance of instigating preventive measures against SSI.

Eccrine angiomatous hamartoma (EAH) is a rare, benign cutaneous lesion characterized histologically by a proliferation of eccrine glands and vascular structures-generally capillaries-in the middle and deep dermis. Sudden enlargement of EAH lesions with or without pain has been noted during puberty and pregnancy and has been attributed to hormonal stimulation. We herein describe a case of EAH that became symptomatic in an adolescent girl. A 13-year-old girl presented with pain associated with a sudden enlargement of a previously asymptomatic swelling on her right second toe. She had an 8-year history of an asymptomatic swelling on her right second toe, and the symptoms appeared approximately 1 year after menarche. Physical examination revealed swelling of the plantar surface of her right second toe. The overlying surface was erythematous with a small amount of fine scales. The biopsied tissue showed a nodular proliferation of eccrine glands intimately admixed with numerous small vessels in the deep dermis and subcutaneous fat tissue. Mucin deposition was present in the stroma surrounding the proliferating eccrine coils and ducts and in the upper dermis. A diagnosis of EAH was made. We suggest that hormonal changes during puberty may have played a role in the rapid growth and pain in the present case.