Mutations in SERPINB7, Encoding a Member of the Serine Protease Inhibitor Superfamily,Cause Nagashima-type Palmoplantar Keratosis

“Nagashima-type” palmoplantar keratosis (NPPK) is an autosomal recessive nonsyndromic diffuse palmoplantar keratosis characterized by well-demarcated diffuse hyperkeratosis with redness, expanding on to the dorsal surfaces of the palms and feet and the Achilles tendon area. Hyperkeratosis in NPPK is mild and nonprogressive, differentiating NPPK clinically from Mal de Meleda. We performed whole-exome and/or Sanger sequencing analyses of 13 unrelated NPPK individuals and identified biallelic putative loss-of-function mutations in SERPINB7, which encodes a cytoplasmic member of the serine protease inhibitor superfamily. We identified a major causative mutation of c.796C>T (p.Arg266^∗) as a founder mutation in Japanese and Chinese populations. SERPINB7 was specifically present in the cytoplasm of the stratum granulosum and the stratum corneum (SC) of the epidermis. All of the identified mutants are predicted to cause premature termination upstream of the reactive site, which inhibits the proteases, suggesting a complete loss of the protease inhibitory activity of SERPINB7 in NPPK skin. On exposure of NPPK lesional skin to water, we observed a whitish spongy change in the SC, suggesting enhanced water permeation into the SC due to overactivation of proteases and a resultant loss of integrity of the SC structure. These findings provide an important framework for developing pathogenesis-based therapies for NPPK.     

Omega-3 polyunsaturated fatty acid has an anti-oxidant effect via the Nrf-2/HO-1 pathway in 3T3-L1 adipocytes

Oxidative stress is produced in adipose tissue of obese subjects and has been associated with obesity-related disorders. Recent studies have shown that omega-3 polyunsaturated fatty acid (ω3-PUFA) has beneficial effects in preventing atherosclerotic diseases and insulin resistance in adipose tissue. However, the role of ω3-PUFA on adipocytes has not been elucidated. In this study, 3T3-L1 adipocytes were treated with ω3-PUFA and its metabolites, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or 4-hydroxy hexenal (4-HHE). ω3-PUFA and its metabolites dose-dependently increased mRNA and protein levels of the anti-oxidative enzyme, heme oxygenase-1 (HO-1); whereas no changes in the well-known anti-oxidant molecules, superoxide dismutase, catalase, and glutathione peroxidase, were observed. Knockdown of nuclear factor erythroid 2-related factor 2 (Nrf-2) significantly reduced EPA, DHA or 4-HHE-induced HO-1 mRNA and protein expression. Also, pretreatment with ω3-PUFA prevented H₂O₂-induced cytotoxicity in a HO-1 dependent manner. In conclusion, treatment with EPA and DHA induced HO-1 through the activation of Nrf-2 and prevented oxidative stress in 3T3-L1 adipocytes. This anti-oxidant defense may be of high therapeutic value for clinical conditions associated with systemic oxidative stress.     

Conformational restriction approach to BACE1 inhibitors II: SAR study of the isocytosine derivatives fixed with a cis-cyclopropane ring

To improve the efficacy of the conformationally restricted BACE1 inhibitors, structural modifications were investigated using two strategies: (a) modification of the terminal aromatic ring and (b) insertion of a spacer between the aromatic rings. In the latter approach, another type of inhibitor 17 bearing an ethylene spacer between two aromatic rings was found to exhibit good BACE1 inhibitory activity, while the corresponding conformationally unrestricted compound 25 showed no activity. This result revealed an interesting effect of a conformational restriction with a cyclopropane ring.     

Nuclear and chloroplast DNA phylogeography reveals Pleistocene divergence and subsequent secondary contact of two genetic lineages of the tropical rainforest tree species Shorea leprosula (Dipterocarpaceae) in South‐East Asia

Tropical rainforests in South‐East Asia have been affected by climatic fluctuations during past glacial eras. To examine how the accompanying changes in land areas and temperature have affected the genetic properties of rainforest trees in the region, we investigated the phylogeographic patterns of a widespread dipterocarp species, Shorea leprosula. Two types of DNA markers were used: expressed sequence tag‐based simple sequence repeats and chloroplast DNA (cpDNA) sequence variations. Both sets of markers revealed clear genetic differentiation between populations in Borneo and those in the Malay Peninsula and Sumatra (Malay/Sumatra). However, in the south‐western part of Borneo, genetic admixture of the lineages was observed in the two marker types. Coalescent simulation based on cpDNA sequence variation suggested that the two lineages arose 0.28–0.09 million years before present and that following their divergence migration from Malay/Sumatra to Borneo strongly exceeded migration in the opposite direction. We conclude that the genetic structure of S. leprosula was largely formed during the middle Pleistocene and was subsequently modified by eastward migration across the subaerially exposed Sunda Shelf.     

Phase Changes in Amylose–Butanol Complex Solution

The phase change for an amylose–butanol complex solution in 10% of dimethylsulfoxide was investigated as a function of temperature. The phase change was determined with measurements of the turbidity, fluorescent depolarization, and viscosity. The phase diagram obtained was qualitatively similar to that for an amylose solution. From the result, the change in solution phase for the amylose–butanol complex is suggested to be similar to that for amylose, i.e., when the solution cools from a higher temperature, amylose molecules in the complex solution change the conformation from a random coil to an interrupted helix, and then separate into two phases. Coacervate particles resulting from the phase separation coalesce with each other to yield precipitates.

An adsorption of uranine on amylose was studied to ascertain its relationship with the fluorescent depolarization method used for detecting phase changes in solution. The result showed that uranine was adsorbed on amylose chains but not on the amylose–butanol complex.     

Release,Regeneration and Mutant Induction of Pleurotus cornucopiae (‘Tamogitake’) Protoplasts

Both enantiomers of aliphatic and aromatic cyanohydrins were hydrolyzed with the aid of Rhodococcus butanica ATCC 21197 to afford optically active α-hydroxy acids. The usefulness of this reaction is demonstrated by the synthesis of optically pure (R)-4-dodecanolide, a defensive secretion of rove beetles, starting from (R)-2-hydroxydecanenitrile.     

Changes in Casein and Egg Albumin due to Reactions with Oxidizing Methyl Linoleate in Dehydrated Systems

Casein and egg albumin were allowed to react with methyl linoleate (ML) at a relative humidity (RH) of 0% or 80% at 50°C for 10 days (protein: ML= 1:0.2 or 1:1, w/w). Changes in the molecular sizes of the reacting proteins were examined by gel filtration and gel electrophoresis. Both proteins showed similar changes, whereas the reaction at RH 80% (protein: ML= 1:1) resulted in insolubilization because of polymerization. Changes in the amino acid residues of the reacting proteins were investigated after acid (6 n HC1) and enzymatic (pepsin-pancreatin, followed by aminopeptidase-prolidase) hydrolyses. Insignificant changes were observed in the amino acid composition of proteins reacted at RH 0%. After reaction at RH 80% (protein: ML =1:1), Lys, His and Met were the only amino acids affected. The percentage loss of these amino acids after acid hydrolysis was Lys (22%), His (41%), Met (9%) for casein and Lys (22%), His (31%), Met (1%) for egg albumin. This percentage loss after enzymatic hydrolysis was Lys (41%), His (49%), Met (94%) for casein and Lys (37%), His (42%), Met (88%) for egg albumin. Some differences between our results and other researchers were also discussed.     

Conjugated Serotonins and Phenolic Constituents in Safflower Seed (Carthamus tinctorius L.)

New three conjugated serotonins were isolated from safflower meal (carthamus tinctorius L.). On the basis of spectral properties and chemical evidence, their structures were determined as N-feruloylserotonin (4), N-(p-coumaroyl)serotonin (5) and N-(p-coumaroyl)serotonin mono-β-d-glucopyranoside (6), The known compounds, 2-hydroxyarctiin (2), matairesinol mono-β-d-glucoside (1) and acacetin (3) were also isolated and identified.     

Moisture Sorption Isotherms of Various Tobaccos

The equilibrium moisture contents of sun-cured (Kroumougrad), flue-cured (Bright Yellow—4) and air-cured (Burley-21 and Matsukawa) tobaccos were measured over a relative humidity range from 5 to 80% at 20°C. The moisture sorption isotherms of tobaccos were of sigmoid type, and classified into two groups. In a lower humidity range below ca. 40% RH, the A group (Kroumougrad and BY-4) had a smaller moisture sorption capacity than B group (Burley-21 and Matsukawa), while in a higher humidity range above ca. 50% RH the former had a larger moisture sorption capacity than the latter. By extracting with water, the moisture content of BY-4 was increased in the lower humidity range, while it decreased in the higher humidity range. However, the moisture content of Matsukawa was scarecely changed by extracting it with water. These results suggest that the differences in equilibrium moisture content with the type of curing were due to the differences in contents of water soluble com- ponents. To control the hygroscopic properties of a tobacco, therefore, the influences of the addition of sucrose and glycerol on the equilibrium moisture content were quantitatively analysed. The moisture sorption capacity of tobacco was greatly different from its nitrogen sorption capacity. The specific surface area of tobacco calculated from moisture sorption isotherm was ca. 110 times larger than the specific surface area calculated from the nitrogen sorption isotherm. Both the nitrogen and moisture sorption data should be necessary for better understanding of the complicated sorption-desorption phenomena in tobaccos.