Development and crystallographic evaluation of histone H3 peptide with N-terminal serine substitution as a potent inhibitor of lysine-specific demethylase 1.

Lysine-specific demethylase 1 (LSD1/KDM1A) is a flavoenzyme demethylase, which removes mono- and dimethyl groups from histone H3 Lys4 (H3K4) or Lys9 (H3K9) in complexes with several nuclear proteins. Since LSD1 is implicated in the tumorigenesis and progression of various cancers, LSD1-specific inhibitors are considered as potential anti-cancer agents. A modified H3 peptide with substitution of Lys4 to Met [H3K4M] is already known to be a potent competitive inhibitor of LSD1. In this study, we synthesized a series of H3K4M peptide derivatives and evaluated their LSD1-inhibitory activities in vitro. We found that substitutions of the N-terminal amino acid with amino acids having a larger side chain were generally not tolerated, but substitution of Ala1 to Ser unexpectedly resulted in more potent inhibitory activity toward LSD1. X-ray crystallographic analysis of H3K4M derivatives bound to the LSD1·CoREST complex revealed the presence of additional hydrogen bonding between the N-terminal Ser residue of the H3 peptide derivative and LSD1. The present structural and biochemical findings will be helpful for obtaining more potent peptidic inhibitors of LSD1.

Activation of lysine-specific demethylase 1 inhibitor peptide by redox-controlled cleavage of a traceless linker.

We have previously employed cyclization of a linear peptide as a strategy to modulate peptide function and properties, but cleavage to regenerate the linear peptide left parts of the linker structure on the peptide, interfering with its activity. Here, we focused on cyclization of a linear peptide via a "traceless" disulfide-based linkage that would be cleaved and completely removed in a reducing environment, regenerating the original linear peptide without any linker-related structure. Thus, the linker would serve as a redox switch that would be activated in the intracellular environment. We applied this strategy to a lysine-specific demethylase 1 (LSD1) inhibitor peptide 1. The resulting cyclic peptide 2 exhibited approximately 20 times weaker LSD1-inhibitory activity than peptide 1. Upon addition of reducing reagent, the linker was completely removed to regenerate the linear peptide 1, with full restoration of the LSD1-inhibitory activity. In addition, the cyclic peptide was far less susceptible to proteolysis than the linear counterpart. Thus, this switch design not only enables control of functional activity, but also improves stability. This approach should be applicable to a wide range of peptides, and may be useful in the development of peptide pharmaceuticals.

Design of New Extraction Surfactants for Membrane Proteins from Peptide Gemini Surfactants.

The development of additional extraction surfactants for membrane proteins is necessary for membrane protein research, since optimal combinations for the successful extraction of target membrane proteins from biological membranes that minimize protein denaturation are hard to predict. In particular, those that have a unique basal molecular framework are quite attractive and highly desired in this research field. In this study, we successfully constructed a new extraction surfactant for membrane proteins, NPDGC12KK, from the peptide-gemini-surfactant (PG-surfactant) molecular framework. The PG-surfactant is a U-shaped lipopeptide scaffold, consisting of a short linker peptide (-X-) between two long alkyl-chain-modified Cys residues and a peripheral peptide (Y-) at the N-terminal side of long alkyl-chain-modified Cys residues. Using photosystem I (PSI) and photosystem II (PSII) derived from Thermosynecoccus vulcanus as representative membrane proteins, we evaluated whether NPDGC12KK could solubilize membrane proteins while maintaining structure and functions. Neither the membrane integral domain nor the cytoplasmic domain of PSI and PSII suffered any damage upon the use of NPDGC12KK based on detailed photophysical measurements. Using thylakoid membranes of T. vulcanus as a representative biological membrane sample, we performed experiments to extract membrane proteins, such as PSI and PSII. Based on the extraction efficiency and maintenance of protein supramolecular structure established using clear native-PAGE analyses, we proved that NPDGC12KK functions as a novel class of peptide-containing extraction surfactants for membrane proteins.

Serum brain-derived neurotrophic factor level in elderly women depression: a community-based study.

OBJECTIVES: Serum levels of brain-derived neurotrophic factor (BDNF) have been shown to be lower in patients with major depressive disorder (MDD) than in healthy persons. Although several studies have examined the associations between serum BDNF levels and broader categories of depression identified by psychiatrists or depressive symptoms measured with depression scales among nonpatient populations, some of these studies did not consider possible confounders and included mostly young or middle-aged subjects and nonrepresentative control subjects, such as volunteers and patients' relatives. Therefore, it remains unclear that whether MDD, broader categories of depression, or depressive symptoms in the elderly are associated with BDNF. The present study examined these associations in a community sample and controlled for confounders. METHODS: The subjects were 538 women aged 78 to 88 years who had participated in a follow-up survey of a cohort and had scored 24 or more on the Mini-Mental State Examination. Two depression scales were administered, and, using the Structured Clinical Interview for DSM-IV, psychiatrists identified 53 persons having any mood disorder (AMD) - 8 with MDD and 45 with other types of depression according to the DSM-IV or its research criteria - and 106 healthy controls. RESULTS: Subjects with MDD had serum BDNF levels lower than did controls but subjects with AMD did not. The severity of depressive symptoms assessed with either of the 2 depression scales was negatively correlated with serum BDNF levels in all subjects and in subjects remaining after persons with MDD or AMD were excluded. These associations were significant after controlling for possible confounders. CONCLUSION: We have found an association between MDD and serum BDNF levels in old-old women, as has previously been found in younger patients. Although serum BDNF levels were not found to be associated with the broader category of depression, they were associated with depressive symptoms among subjects without clinical depression.

Sake lees fermented with lactic acid bacteria prevents allergic rhinitis-like symptoms and IgE-mediated basophil degranulation.

We tested the effect of oral administration of fermented sake lees with lactic acid bacteria (FESLAB) on a murine model of allergic rhinitis upon immunization and nasal sensitization with ovalbumin (OVA). We used Lactobacillus paracasei NPSRIk-4 (isolated from sake lees), and L. brevis NPSRIv-8 (from fermented milk) as starter strains to produce the FESLAB. Oral FESLAB administration resulted in the development of significantly fewer sneezing symptoms than those seen in sham control animals given sterile water. We also found that FESLAB suppressed the allergen-induced degranulation of RBL2H3 rat basophilic leukemia cells.

[Nonpharmacologic treatment for insomnia].

Nonpharmacologic treatment for insomnia occupies an important position on the treatment of insomnia. It is necessary to educate the patient on sleep hygiene to adjust the unappropriate sleep pattern. Within methods of relaxation, autonomy training and biofeedback method are methods using muscle relaxation to achieve psychological balance. Morita therapy is a method to accept insomnia as it is, and give guidance on the patient's life activity in a constructive way. It is important to listen carefully to the patient's life background and choose treatment based on that information. For this, an appropriate construction of a doctor-patient relationship is essential.

Stress and psychological factors before a migraine attack: a time-based analysis.

BACKGROUND: The objective of this study is to examine the stress and mood changes of Japanese subjects over the 1-3 days before a migraine headache. METHODS: The study participants were 16 patients with migraines who consented to participate in this study. Each subject kept a headache diary four times a day for two weeks. They evaluated the number of stressful events, daily hassles, domestic and non-domestic stress, anxiety, depressive tendency and irritability by visual analog scales. The days were classified into migraine days, pre-migraine days, buffer days and control days based on the intensity of the headaches and accompanying symptoms, and a comparative study was conducted for each factor on the migraine days, pre-migraine days and control days. RESULTS: The stressful event value of pre-migraine days showed no significant difference compared to other days. The daily hassle value of pre-migraine days was the highest and was significantly higher than that of buffer days. In non-domestic stress, values on migraine days were significantly higher than on other days, and there was no significant difference between pre-migraine days and buffer days or between pre-migraine days and control days. There was no significant difference in the values of domestic stress between the categories. In non-domestic stress, values on migraine days were significantly higher than other days, and there was no significant difference between pre-migraine days and buffer days or between pre-migraine days and control days.There was little difference in sleep quality on migraine and pre-migraine days, but other psychological factors were higher on migraine days than on pre-migraine days. CONCLUSION: Psychosocial stress preceding the onset of migraines by several days was suggested to play an important role in the occurrence of migraines. However, stress 2-3 days before a migraine attack was not so high as it has been reported to be in the United States and Europe. There was no significant difference in the values of psychological factors between pre-migraine days and other days.