Effects of permanent waving on changes of protein and physicomorphological properties in human head hair.

Two types of permanent waving [digital perm (DP) and croquignole winding perm (CWP)] and two waving lotions [cysteamine-HCl, pH 9.31, liquid type (lotion A) and sodium thioglycolate, pH 9.97, cream type (lotion B)] were used for this study. The protein content was decreased by permanent waving treatments on the whole, and the degree of reduction was dependent on the hair styling and waving lotion used. The greatest decrease (by 58%) was found in hairs processed with the three-treatment performance of DP using lotion B. SDS-PAGE identified the presence of the two most abundant polypeptides, with approximately 48 kDa and 60 kDa, and two large polypeptides, with approximately 200 kDa and 210 kDa, which would belong to the keratin family. Some physical properties of the hairs (tensile strength, diameter, swelling, and elongation) were examined with the permanent waving treatments. In general, the repeated waving treatment and the use of lotion B showed more negative effects on hair care than other treatments. Some morphological changes were observed with a scanning electron microscope (SEM). The most prominent change in the hair surface was observed in the hair specimen with the three-treatment performance of DP using lotion B. More severe signs of damage appeared on the hair with lotion B than with lotion A. As the numbers of permanent waves increased, the degree of damage to the hair surface increased on the whole. However, there was no indication of changes to the hair surface with one permanent waving treatment, as determined by SEM analysis.

A simple improved method for protein extraction from human head hairs.

This study was conducted to establish a simple efficient method for extracting the protein from human head hair materials, which can be a useful tool for the protein analysis applicable to various types of human head hairs. The method developed saves extraction time and effort considerably. The method includes four steps: cutting the hair samples into small pieces 1-2 mm in length, washing them with distilled water, incubating the hair samples in a buffer solution at 50 degrees C for 24 hr, and finally filtering the incubated mixtures through three layers of nylon mesh. This method is reproducible and reliable. SDS-PAGE analysis of the hair protein extracted by this method shows a clear protein profile on the gel, which is frequently observed in other hair sources. Two smaller sizes of molecular weights are also detected with the SDS-PAGE analysis. Not commonly found in other hair sources, they seem to be other types of human hair proteins.

Molecular and biochemical characterizations of dehydroascorbate reductase from sesame (Sesamum indicum L.) hairy root cultures.

Dehydroascorbate reductase (DHAR) is a biotechnologically or physiologically important reducing enzyme in the ascorbate-glutathione recycling reaction for most higher plants. A DHAR cDNA was isolated from sesame (Sesamum indicum L.) hairy roots, and its structure and biochemical properties were characterized to provide some information about its expressional and biochemical profiles in the hairy root cultures. The cDNA contained a catalytic motif CXXS, which may be indicative of a thiol-dependent redox function. A fusion DHAR expressed in an Escherichia coli expression system was purified with four purification steps until a homogeneous single band signal was seen in an acrylamide gel, and its antibody was prepared for Western blot analyses. The biochemical results showed that the purified recombinant DHAR had an optimal pH of around 6.0, which was different from those (pH 7.8-8.2) of other plant species. The temperature optimal for the DHAR activity was in a relatively wide range of 30-60 degrees C. It was proved by a real-time RT-PCR technique that the transcription activity of the DHAR was about 2-5-fold higher during the first 3 week cultures than during the latter 3 week ones. The highest activity of the sesame DHAR was detected in the 4 week cultures of the hairy roots, after which its activity was rapidly decreased to approximately 80%, suggesting that the most active DHAR occurred in this culture period. Western blot analyses confirmed that the presence of DHAR enzyme was identified in both cultures of the fused E. coli and the sesame hairy roots.

Expression and characterization of extracellular fungal phytase in transformed sesame hairy root cultures.

A recombinant fungal phytase was produced by cultures of sesame hairy roots transformed with Agrobacterium rhizogenes, purified and its molecular properties were characterized. Its transcription level and the phytase production were rapidly increased after 4 weeks of the cultures, suggesting that its transcription and protein synthesis might concur. Western blot analysis provided evidence that the recombinant fungal phytase was secreted into the liquid culture medium of the hairy roots. The phytase enzyme secreted was purified by three steps of ultrafiltration, DEAE-Sepharose ion exchange chromatography, and Sephadex G-100 size-exclusion chromatography. As a result, one single band signal was observed with SDS-PAGE, indicating that the purification step was reasonable. The positive signs of both the zymogram and the PAS staining on SDS-PAGE suggested that the activity of the final product phytase was active and glycosylated. The optimal reaction temperature of the phytase was between 50 and 60 degrees C and at over 60 degrees C its activity was reduced by 30-90%, depending on the temperatures applied. Pre-incubation at temperatures of 20-50 degrees C showed stable catalytic activity, while at over 50 degrees C the phytase activity was gradually decreased by 90%. The optimal pH was between 4 and 5 pH values for the recombinant fungal phytase, while for native phytase it was at pH 5.0. Addition of iron ion inhibited the phytase activity but treatments of some cations, EDTA, and PMSF showed no effect on the activity or slightly stimulated it positively.

Isolation and characterization of a myo-inositol 1-phosphate synthase cDNA from developing sesame (Sesamum indicum L.) seeds: functional and differential expression, and salt-induced transcription during germination.

A cDNA (SeMIPS1) encoding myo-inositol 1-phosphate synthase (EC 5.5.1.4) (MIPS) has been characterized from sesame (Sesamum indicum L. cv. Dan-Baek) seeds and its functional expression analyzed. The SeMIPS1 protein was highly homologous with those from other plant species (88-94%), while a much lower degree of sequence homology (53-62%) was found with other organisms such as humans, mouse, algae, yeast, Drosophila, bacteria and other prokaryotes. A yeast-based complementation assay in yeast mutants containing a disrupted INO1gene for yeast MIPS confirmed that the SeMIPS1 gene encodes a functional MIPS. Phylogenetic analysis suggested that the SeMIPS1 gene diverged as a different subfamily or family member. Southern hybridization revealed several copies of the SeMIPS1 gene present in the sesame genome and northern blotting indicated that expression of the SeMIPS1gene may be organ specific. Salt stress during sesame seed germination had an adverse influence on transcription of SeMIPS1and greatly reduced transcript levels as the duration of exposure to a saline environment increased and NaCl concentration increased. Germination initiation of sesame seeds was severely delayed as NaCl level increased. These results suggest that expression of SeMIPS1 is down-regulated by salt stress during sesame seed germination.