Unusual reactivity of ß-hydroxy-serotonin, a forgotten serotonin derivative.

Serotonin (5-HT) is a well-known biogenic amine which regulates mood, sleep, and is involved in muscle contraction and blood coagulation. Based on an analogy to norepinephrine, a ß-hydroxylated derivative of dopamine which has diverse physiological functions, beta-hydroxy-serotonin (ß-OH-5-HT) originally encouraged interest as a potential pharmacological agent. Four decades ago, its organic synthesis was attempted. However, due to difficulties with the synthesis and the compound's instability, rigorous identification and characterization of ß-OH-5-HT proved evasive. Here, we successfully synthesized ß-OH-5-HT from 5-HT using a Pseudomonas enzyme, tryptophan side chain oxidase type I (TSOI), and we determined the structure by 2D-NMR and characterized ß-OH-5-HT in detail. The CD spectra showed no optical activity, suggesting a racemic mixture. To separate DL-ß-OH-5-HT, we synthesized L-Ala-5-HT and derivatized it into erythro- and threo-L-Ala-ß-OH-5-HT with TSOI. Interestingly, both isolated fractions returned to a diastereoisomeric mixture within two hours at pH 5.0. Later, we found that, under acidic conditions, ß-OH-5-HT readily reacted with nucleophiles like alcohols or thiols, yielding a variety of DL-ß-substituted-5-HT. The unusual properties of ß-OH-5-HT might be attributed to the unique nature of a ß-hydroxyl group adjacent to an indole ring and amino group. The mechanism for the rapid racemization of ß-OH-5-HT is discussed.

Downregulation of trefoil factor 3 gene expression in the colon of the senescence-accelerated mouse (SAM)-P6 revealed by oligonucleotide microarray analysis.

Global comparison of the colonic gene expression profiles between 14-month-old senescenceaccelerated mouse (SAM)-P6 mice and SAM-R1 mice, a wild-type control, was conducted with an oligonucleotide microarray containing more than 5,000 mouse genes. Eight genes were upregulated more than two-fold and 94 genes were downregulated more than two-fold in SAM-P6 mice. The three cell defense genes intelectin1 (Itln1), trefoil factor 3 (intestinal) (Tff3) and "deleted in malignant brain tumors 1" (Dmbt1) were among those extensively downregulated. Quantitative RT-PCR analysis confirmed that Itln1 mRNA was almost undetectable in SAM-P6 colon, whereas it was readily detected in SAM-R1 colon. Colonic expression of both Tff3 and Dmbt1 mRNA was also substantially decreased, to one third and two thirds of the levels in SAM-R1 mice, respectively. A 14 kDa Tff3 dimer was detected by Western blotting in the colon of all three SAM-R1 mice, but was not present in three SAM-P6 mice. No upregulation of 3 cell defense genes was detected in 3-month-old SAM-R1 as well as SAM-P6 mice. These results suggest that a diminution of the intestinal trefoil factor system may be involved in the acceleration of aging in SAM-P6 mice.

Composition of milk obtained from unmassaged versus massaged breasts of lactating mothers.

BACKGROUND: The Oketani method is a program of breast massage and clinical counseling developed by the midwife Satomi Oketani. The purpose of this study is to examine the effects of the method on the quality of breast milk by determining the chemical composition of the milk before and after massage. METHODS: Milk samples were obtained immediately before and after massage from healthy, exclusively breast-feeding Japanese mothers at two different periods of lactation one <3 months the other >3 months after parturition. Lipids, whey protein, casein, lactose, ash, and total solids in milk were measured in milk samples. The gross energy content of milk was estimated. RESULTS: Breast massage significantly increased lipids in the late lactating period but not in the early lactating period. In the early lactating period casein was increased by breast massage but was not significantly affected in the late lactating period. Breast massage caused a significant increase in total solids from the first day to 11 months post partum. The gross energy in the late lactating period was significantly increased by breast massage but not in the early lactating period. Lactose was not significantly changed by breast massage. CONCLUSIONS: Breast massage improves the quality of human milk by significantly increasing total solids, lipids, and casein concentration and gross energy. The milk of mothers treated by Oketani breast massage may improve the growth and development of infants.