Hair growth promoting potential of phospholipids purified from porcine lung tissues.

BP201, porcine lung tissue-derived phospholipids, consists of phosphatidylcholine as a major phospholipid species. BP201 promoted hair growth after application onto the shaved backs of BALB/c and C3H mice. Its effect was enhanced when applied together with minoxidil (MNX) in C3H mice. When the tissue specimens prepared from the shaved skins of BP201-treated and control mice were microscopically examined, the total numbers of hair follicles in both anagen and telogen phases of BP201-treated mice were significantly higher than those of control mice. The numbers of hair follicles in the anagen phase of BP201-treated mice were also higher than those of control mice. In combination with MNX, BP201 further increased the total number of hair follicles, but did not alter the percentage of hair follicles in the anagenic phase. BP201 also increased the proliferation of human hair follicle dermal papilla cells. Collectively, BP201 possesses hair growth promoting potential, which would suggest its use singly or in combination for hair growth products.

Topical application of a phospholipid mixture purified from pig lungs ameliorates 2,4-dinitrofluorobenzene-induced allergic contact dermatitis in BALB/c mice.

This work was designed to assess the pharmacological effectiveness as a novel anti-atopic dermatitis remedy of a phopholipid mixture purified from pig lung tissues, named KT&G101, using the BALB/c mouse model of allergic contact dermatitis. Allergic contact dermatitis was induced by applying 2,4-dinitrofluorobenzene (DNFB) epicutaneously onto the dorsal skins of mice, and KT&G101 was topically applied onto the skin areas with the lesions. The topical application of KT&G101 (0.05 ml of 10 mg/ml and 20 mg/ml KT&G101, twice a day for 15 days) decreased the total IgE level elevated in the sera of mice undergoing allergic contact dermatitis. KT&G101 was also able to decrease the 2,4-dinitrophenyl (DNP)-specific IgE level elevated in the sera of the model mice. It reduced the incidences of scratching behaviors in the mice undergoing DNFB-induced allergic contact dermatitis. It attenuated some histopathological changes, such as pustule, epidermal hyperplasia, dermatitis and fibroplasia, while it could enhance the recovery of epidermis, in the damaged skin tissues within a relatively short period after the topical application of KT&G101. KT&G101 lessened the expression of cytokines mRNAs, such as Th1-specific IL-2, TNF-ß and IFN-γ, and Th2-specific IL-4, in the mouse skin tissues showing the lesions. In brief, it is concluded that KT&G101 alleviates the symptoms involved in induced allergic contact dermatitis in BALB/c mice.

Anti-inflammatory, antinociceptive and anti-angiogenic activities of a phospholipid mixture purified from porcine lung tissues.

This work aimed to assess anti-inflammatory and related properties of a phospholipid mixture purified from porcine lung tissues, named KT&G101, which is being developed as a novel topical remedy for atopic dermatitis. KT&G101 consists of pure phospholipids, mainly phosphatidylcholine (PC) and other phospholipids such as phosphatidylinositol (PI) and phosphatidylserine (PS). Its predominant PC species is 1,2-dipalmitoylphosphatidylcholine (DPPC). KT&G101 exhibited an anti-angiogenic activity in the chick chorioallantoic membrane (CAM) assay. Oral administration of KT&G101 at the dosages of 100, 200 and 400 mg/kg body weight gave rise to an inhibition of 15.4%, 25.3% and 30.1% in the vascular permeability assay, respectively. In the carrageenan-induced inflammation in the air pouches, KT&G101 significantly diminished the volume of exudates in the pouches, the number of polymorphonuclear leukocytes and nitrite content in exudates. In the acetic acid-induced writhing response, oral administration of KT&G101 at the dosages of 50, 100 and 200 mg/kg body weight showed the reduction of 21.6%, 51.6% and 60.8% in the pain response of mice, respectively. It was also able to diminish the nitric oxide (NO) and reactive oxygen species (ROS) levels in the lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. KT&G101 displayed a significant suppression on the induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the stimulated RAW264.7 cells. However, the free radical scavenging activity of KT&G101 was detected to be very weak in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Taken together, KT&G101 possesses anti-inflammatory and related antinociceptive and anti-angiogenic activities, which indirectly supports its use as an anti-atopic therapy.

Characterization and regulation of the gene encoding monothiol glutaredoxin 3 in the fission yeast Schizosaccharomyces pombe.

Glutaredoxins (Grxs) are thioloxidoreductases which are required for maintaining thiol/disulfide equilibrium in living cells. The Grx3 gene, which encodes one of the three monothiol Grxs in the fission yeast Schizosaccharomyces pombe, was characterized, and its transcriptional regulation studied. Genomic DNA encoding Grx3 was isolated by PCR, and a plasmid pTT3 carrying this DNA was produced. The DNA sequence has 1,267 bp, which would encode a monothiol Grx of 166 amino acids with a molecular mass of 18.3 kDa. The putative protein has 27% homology with Grx5, and contains many hydrophobic amino acid residues in its N-terminal region. S. pombe cells harboring pTT3 had increased Grx activity and enhanced survival on minimal medium plates containing aluminum (5 mM), BSO (0.05 mM), menadione (0.01 mM) or cadmium (0.2 mM). The 568 bp upstream region of Grx3 was fused into the promoterless beta-galactosidase gene of the shuttle vector YEp367R to generate fusion plasmid pMJS10. Potassium chloride (KCl) and metals including aluminum and cadmium enhanced the synthesis of beta-galactosidase from the fusion gene. The synthesis of beta-galactosidase was also enhanced, in a Pap1-dependent manner, by fermentable carbon sources such as glucose (at low concentrations) and sucrose, but not by non-fermentable carbon sources such as ethanol and acetate. Grx3 mRNA increased in response to treatment with BSO. These observations indicate that S. pombe Grx3 is involved in the response to stress, and is regulated by stress.

Characterization of a second gene encoding gamma-glutamyl transpeptidase from Schizosaccharomyces pombe.

The first gene encoding gamma-glutamyl transpeptidase (GGTI) of the fission yeast has previously been characterized, and its expression was found to be regulated by various oxidative stress-inducing agents. In this work, a second gene, encoding GGTII, was cloned and characterized from the fission yeast Schizosaccharomyces pombe. The structural gene encoding GGTII was amplified from the genomic DNA of the fission yeast and ligated into the shuttle vector pRS316 to generate the recombinant plasmid pPHJ02. The determined sequence contains 3040 bp and is able to encode the putative 611 amino acid sequence of GGTII, which resembles the counterparts of Saccharomyces cerevisiae, Homo sapiens, Rattus norvegicus, and Escherichia coli. The DNA sequence also contains 940-bp upstream and 289-bp downstream regions of the GGTII gene. The Schizosaccharomyces pombe cells harboring plasmid pPHJ02 showed about 4-fold higher GGT activity in the exponential phase than the cells harboring the vector only, indicating that the cloned GGTII gene is functional. The S. pombe cells containing the cloned GGTII gene were found to contain higher levels of both intracellular glutathione (GSH) content and GSH uptake. The S. pombe cells harboring plasmid pPHJ02 showed increased survival on solid media containing hydrogen peroxide, diethylmaleate, aluminum chloride, cadmium chloride, or mercuric chloride. The GGTII mRNA level was significantly elevated by treatment with GSH-depleting diethylmaleate. These results imply that the S. pombe GGTII gene produces functional GGTII protein and is involved in the response to oxidative stresses in S. pombe cells.

Age-related changes in the activity of antioxidant and redox enzymes in rats.

Cellular defense system, including glutathione, glutathione-related enzymes, and antioxidant and redox enzymes, may play crucial roles in the aging of aerobic organisms. To understand the physiological roles of these factors in the aging process, their levels were compared in the livers and brains of 5-week- and 9-month-old rats. GST activity was higher in livers and brains of 9-month-old rats than in those of 5-week-old rats, and brain catalase activity was about 2-fold higher. However, it was unchanged in the livers of the 9-month-old rats. gamma-Glutamylcysteine synthetase activity was about 2-fold higher in the brains of the older rats but again not in their livers. In contrast glutathione synthetase activity appeared to be lower in the livers of the older rats while GSH content did not change with age in livers and brains. Glutathione peroxidase activity was higher in 9-month-old rat brains, but lower in 9-month-old rat livers, while superoxide dismutase activity was higher in both tissues in the older rats. The activities of two redox enzymes, thiol-transferase and thioredoxin reductase, did not change with age, nor did that of glutathione reductase. These results indicate that levels of different cellular defense systems vary with age in an irregular manner.