A Novel Fluorescent GFP Chromophore Analog-Based Dye for Quantitative PCR.

This is the first report describing the possibility of using a green fluorescent protein chromophore synthetic analog, P-HOBDI-BF2, as a fluorescent dye for a linear hydrolysis probe used in qPCR. The study was carried out on a system for detection of the plant pathogenic fungus Fusarium avenaceum using a plasmid containing translation elongation factor 1α fragment as a template. To estimate fluorogenic properties of P-HOBDI-BF2, 6-FAM- and BDP-FL-labeled probes were used. It was demonstrated that a synthetic dye based on the P-HOBDI-BF2 chromophore can be used for labeling hydrolysis probes for qPCR, but fluorescence increase levels for P-HOBDI-BF2-labeled probes were slightly lower than those for 6-FAM-labeled ones. At the same time, the sensitivity of P-HOBDI-BF2-based assays remained high, and this fact together with acceptable fluorescence levels suggests that this dye can be considered as an efficient alternative for reporters traditionally used for fluorescence detection in the FAM channel.

[Novel Water-Soluble Substrate for Silicateins].

We suggested to use tetrakis(2-hydroxyethyl)orthosilicate (THEOS) as a substrate for silicateins--an enzyme family playing a key role in formation of skeleton in marine sponges. We compared THEOS with tetraethylorthosilicate (TEOS)--a commonly used substrate for silicateins. These substrates were tested in reaction of amorphous silica formation in vitro catalyzed by silicatein Al from sponge Latrunculia oparinae. It was found that reaction with THEOS occurs more efficiently than with TEOS, probably due to high water solubility and higher hydrolysis rate of THEOS.

[Synthesis of the chromophores of fluorescent proteins and their analogs].

Members of the green fluorescent protein (GFP) family are widely used in experimental biology as genetically encoded fluorescent tags. Chromophores of GFP-like proteins share a common structural core: 3,5-dihydro-4H-imidazol-4-one. This review covers synthetic approaches to 3,5-dihydro-4H-imidazol-4-ones, substituted at different positions. General, as well as specific methods, represented by single examples are considered. The most popular synthetic route to substituted 3,5-dihydro-4H-imidazol-4-ones includes synthesis of azlactones, followed by transformation into N-acyldehydroaminoacids and, finally, cyclization into target heterocycles. Accordingly, the review is divided into three parts: the first part covers syntheses of azlactones, the second part covers main approaches to N-acyldehydroaminoacids, and in the third part we summarize cyclizations of N-acyldehydroaminoacids, as well as all other approaches to 3,5-dihydro-4H-imidazol-4-ones.

The impact of the April 1992 civil unrest on the Los Angeles REI WIC program and its participants.

This paper discusses the findings of a study conducted in south central Los Angeles in August 1992 among women in the Special Supplemental Food Program for Women, Infants, and Children. The goals of the study were to determine the current demographics of WIC participants; examine the financial hardship, need for relief services, and extent of hunger resulting from the civil unrest of April 1992; look at the effects of the unrest on different ethnic groups; determine the unmet need for WIC services; and evaluate the State and local WIC responses to the unrest. The 1,189 respondents were approximately 77 percent Latina, 20 percent African American, and 3 percent white. Half or more were recent immigrants, 19 percent were pregnant and parenting adolescents, 74 percent were school dropouts, and 56 percent were single mothers. Only 1 percent had any problems using WIC vouchers after the unrest, although more than half of their grocery stores had closed. Thirty-five percent experienced food deficits in their households, and 33 percent of those who applied for emergency food stamps had trouble getting them. Four percent said their children had gone to bed hungry in the last week, and 9 percent said they, the respondents, had as well. Only 2 percent needed shelter, and 1 percent became homeless, but 6 percent had family members who lost jobs due to the unrest. This study suggests that the chronically substandard conditions under which many families in south central Los Angeles live affect them more profoundly than did the dramatic consequences of the civil unrest.