Prooxidant action of gallic acid compounds: copper-dependent strand breaks and the formation of 8-hydroxy-2'-deoxyguanosine in DNA.

Gallic acid and its alkylesters, polyphenolic compounds with antioxidative activity, acted as a prooxidant causing a copper-dependent DNA damage. Treatment of DNA from plasmid pBR322 and calf thymus with gallic acid plus copper ion caused strand scission and the formation of 8-hydroxy-2'-deoxyguanosine in DNA. Addition of catalase protected DNA from the gallic acid/copper-dependent strand breaks and the formation of 8-hydroxy-2'-deoxyguanosine, indicating that hydroxyl radical may participate in the DNA damage. Ethyl-, propyl- and butylgallates showed only a little DNA damage. Octyl- and laurylgallates caused negligible damage of DNA. DNA strand breaks and formation of 8-hydroxy-2'-deoxyguanosine were closely related to the reduction of copper by gallate compounds. These results imply that cuprous ion reduced by gallate derivatives may play a key role in the oxidative cleavage of DNA and the formation of base adduct. The cytotoxic effect of gallate compounds can be explained by their prooxidant action dependent on the reducing activity.

Aluminum-induced apoptosis in PC12D cells.

The addition of aluminum-maltol complex to PC12D cells induced a time-dependent and concentration-dependent growth inhibition as well as cell death, whereas aluminum chloride or maltol alone did not affect the viability of PC12D cells. Apoptosis of differentiated PC12D cells was assessed by using terminal deoxynucleotidyltransferase-mediated 2'-deoxyuridine-5'-triphosphate nick end labeling (TUNEL) technique to detect DNA strand breaks in situ. The number of TUNEL-positive cells treated with aluminum-maltol increased with time in the treatment cultures. The ability of aluminum ion to elevate intracellular reactive oxygen species was determined by fluorescence in PC12D cells loaded with the oxidant-sensitive dye 2',7'-dichlorofluorescin diacetate. Aluminum ion incorporated to PC 12D cells causes apoptotic cell death by enhancing the generation of reactive oxygen species.

Field trials of a rapid test for G6PD deficiency in combination with a rapid diagnosis of malaria.

A rapid single-step screening method for detection of glucose-6-phosphate dehydrogenase (G6 PD) deficiency was evaluated on Halmahera Island, Maluku Province, Indonesia, and in Shan and Mon States, Myanmar, in combination with a rapid diagnosis of malaria by an acridine orange staining method. Severe deficiency was detected by the rapid test in 45 of 1126 volunteers in Indonesia and 54 of 1079 in Myanmar, but it was difficult to distinguish blood samples with mild deficiency from those with normal activity. 89 of 99 severely deficient cases were later confirmed by formazan ring method in the laboratory, but 5 with mild and 5 with no deficiency were misdiagnosed as severe. Of the samples diagnosed as mild and no deficiency on-site, none was found to be severely deficient by the formazan method. Malaria patients were simultaenously++ detected on-site in 273 samples on Halmahera island and 277 samples from Shan and Mon States. In Mon State, primaquine was prescribed safely to G6 PD-normal malaria patients infected with Plasmodium vivax and/or gametocytes of P. falciparum. The new rapid test for G6 PD deficiency may be useful for detecting severe cases under field conditions, and both rapid tests combined are can be useful in malaria-endemic areas, facilitating early diagnosis, prompt and radical treatment of malaria and suppression of malaria transmission.

Identification of human urinary stains by the quotient uric acid/urea nitrogen.

Uric acid (UA) and urea nitrogen (UN) were determined in urinary stains and the UA/UN x 20 values were calculated. The values in human urinary stains were 1.11-4.21, while those in other mammals except some of chimpanzees, were under 0.7, and those in fecal stains of birds were over 80. Most of the stains of other human body fluids or plant juices tested contained neither UA nor UN, and some contained one, but never the other. Ascorbic acid (AS) of up to 100 mg/dl in urine did not interfere with UA determination when dried human urinary stains were analyzed. It was also found that the contents of UA were very low at the peripheral parts of urinary stains. The present results indicate that the quotient UA/UN is useful for identification of human urinary stains in forensic practice provided that the peripheral part of the stain is not used.

Comparative immunology of mammalian uricase using anti-hog liver uricase.

1. Uricase was partially purified from livers of five mammals (dog, bovine, horse, house musk shrew and guinea pig), and its immunological properties were studied using anti-hog liver uricase prepared in rabbits. 2. Results obtained by immunodiffusion suggest that hog uricase has four sites of partial antigens (a, b, c, d), while the enzyme of other mammals has only part of them, and a cladogram of these six mammals is presented showing development of partial antigens. 3. Results obtained by immunoelectrophoresis suggest that the protein structure of house musk shrew uricase is considerably different from the other five mammals tested.

Antigenic properties of human and animal bloodstains studied by enzyme-linked immunosorbent assay (ELISA) using various antisera against specific plasma proteins.

Antigenic properties of bloodstains of human and non-human primates as well as other animal bloodstains were investigated by the inhibition ELISA using commercially available anti-human albumin (Alb), alpha 2-macroglobulin (alpha 2-M), fibrinogen, transferrin, and immunoglobulin G. In general, chimpanzee bloodstains showed strong cross-reactions with these antisera, and the extent of the cross-reactions of other animal bloodstains decreased largely with the phylogenic order, i.e., agile gibbon (ape), Old World monkeys (Japanese monkey and hamadryas baboon), New World monkeys (night monkey and tufted capuchin monkey), prosimians (grand galago and ring-tailed lemur) and other animals (rat, cattle, swine, goat, dog, cat, and chicken). Among these antisera, anti-human alpha 2-M showed the weakest cross-reaction with chimpanzee bloodstains, and anti-human Alb showed next.