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Research into the sustainability of natural, potentially renewable, resources is one of the major issues of our time. It naturally includes the quest for sustainable sources of colorants for textiles, cosmetics, and food. In industrialized countries, natural dyeing with plants and a few species of coccid insects was practiced on a large scale for centuries before synthetic colorants were developed. Therefore, historical documents on the growing of dye plants and dyeing processes offer a relevant basis from which to start reconsidering the potential of natural colorants in our time. However, written sources need to be completed by experimental archaeologists to allow a scientific understanding of the biochemical reactions at work in the historical processes described. The results of such interdisciplinary research can then inspire contemporary programs to revive the production of natural dyes. The long history of dyeing blue with woad, Isatis tinctoria L., is revisited here as an illustration of the fruitful complementarity of sources and approaches. This article presents a step-by-step re-assessment of the production chain of woad as described in historical texts, from the growing of the plant to its use as a source of indigo in the woad and indigo vats. The experimental reconstitution of the processing of woad leaves into couched woad allowed us to follow the evolution of the composition and proportions of indigoid colorants in the leaves by HPLC analyses. Additionally, HPLC analyses allowed a comparison of the respective indigoid contents of couched woad and sukumo, the form of indigo dye resulting from another couching process, traditionally used in Japan for dyers’ knotweed, Persicaria tinctoria (Ait.) H. Gross. The reconstitution of the 18th century woad and indigo vat process allowed investigations into the bacterial flora associated with the use of couched woad in vat liquors, which were found to contain different indigo-reducing bacteria, including two distinct strains of a new indigo-reducing species.

Water-insoluble indigo is solubilized by the reducing action of microorganisms which occurs during fermentation. In natural indigo fermentation, composted leaves of Polygonum tinctorium L. (sukumo) are the raw material that has been used as both the indigo source and the bacterial inoculum. Ideally, indigo reduction occurs shortly after preparation of the fermentation vat. The time-to-reduction depends on the quality of the sukumo and the methods for preparation and management of the fermentation batch. We estimated the effect of adding Indigofera tinctoria L. leaf powder (LP) to indigo fermentation in two fermentations originally exhibiting either rapid or slow time-to-reduction (T-sukumo and D-sukumo, respectively). Alkalihalobacillus spp. (97.7%–98.4% similarities with Alkalihalobacillus macyae) were observed only in the LP-added T-sukumo fermentation liquor. They appeared from day 1 (0.7%) and increased to 24.4% on day 6, and their presence was related to indigo reduction. Differences in functional ratio between LP-added and its control batches revealed enhancement of pathways related to reconstitution of cellular functions and substrate metabolisms, to all of which Alkalihalobacillus spp. contributed intensively. In D-sukumo batch, appearance of bacteria necessary to initiate indigo reduction (principally Anaerobacillus/Polygonibacillus) was comparatively slower. LP promotes earlier indigo reduction in both T- and D-sukumo-based batches, owing to its promotion of microbiota transition. The effect of the LP was intensified from day 1 to day 2 in both sukumo using batches according to the assumed function of the microbiota. The initial effect of LP on the T-sukumo batches was more intense than that in the D-sukumo batches and was continued until day 3, while the duration in the T-sukumo batches was continued until day 5. Based on these observations, we propose that the LP functions through its phytochemicals that eliminate oxygen, stimulate the microbiota, and accelerate its transitional changes toward a suitable function that opens the pathway for the extracellular electron transfer using carbohydrates as a substrate.

Water-insoluble indigo is solubilized by the reducing action of microorganisms which occurs during fermentation. In natural indigo fermentation, composted leaves of Polygonum tinctorium L. (sukumo) are the raw material that has been used as both the indigo source and the bacterial inoculum. Ideally, indigo reduction occurs shortly after preparation of the fermentation vat. The time-to-reduction depends on the quality of the sukumo and the methods for preparation and management of the fermentation batch. We estimated the effect of adding Indigofera tinctoria L. leaf powder (LP) to indigo fermentation in two fermentations originally exhibiting either rapid or slow time-toreduction (T-sukumo and D-sukumo, respectively). Alkalihalobacillus spp. (97.7%-98.4% similarities with Alkalihalobacillus macyae) were observed only in the LP-added T-sukumo fermentation liquor. They appeared from day 1 (0.7%) and increased to 24.4% on day 6, and their presence was related to indigo reduction. Differences in functional ratio between LP-added and its control batches revealed enhancement of pathways related to reconstitution of cellular functions and substrate metabolisms, to all of which Alkalihalobacillus spp. contributed intensively. In D-sukumo batch, appearance of bacteria necessary to initiate indigo reduction (principally Anaerobacillus/Polygonibacillus) was comparatively slower. LP promotes earlier indigo reduction in both T- and D-sukumo-based batches, owing to its promotion of microbiota transition. The effect of the LP was intensified from day 1 to day 2 in both sukumo using batches according to the assumed function of the microbiota. The initial effect of LP on the T-sukumo batches was more intense than that in the D-sukumo batches and was continued until day 3, while the duration in the T-sukumo batches was continued until day 5. Based on these observations, we propose that the LP functions through its phytochemicals that eliminate oxygen, stimulate the microbiota, and accelerate its transitional changes toward a suitable function that opens the pathway for the extracellular electron transfer using carbohydrates as a substrate. [ABSTRACT FROM AUTHOR]

The aim of the present study is to evaluate the association of serum uric acid (UA) levels with the risk of incident hypertension among different age groups in men and women using a single large Japanese general cohort. The present study is based on annual health check-up program in Gunma, Japan. We studied 12,029 participants (mean age, 48 ​± ​9 years old; 31% women) free of prevalent cardiovascular disease and hypertension at baseline (2009). Hypertension was defined by self-report, hypertensive medication use, or measured BP ​> ​140/90 ​mmHg ​at each visit. Discrete proportional hazards regression model was used to evaluate the association of UA level at baseline with incident hypertension through 2012 adjusted for age, gender, baseline blood pressure, and other CVD risk factors among different age decade groups in men and women. During follow-up of 3 years, 12% of the cohort (n ​= ​1457) developed hypertension. UA was strongly associated with incident hypertension in the multivariable model in all participants. In age-stratified analysis, participants below 50 years of age in men had the significant association of UA with incident hypertension, whereas participants above 50 years did not. In women, participants above 40 years had the significant association, whereas participants below 40 years did not. The present data suggest that UA level is an independent predictor for incident hypertension among middle aged men below 50 years old and middle aged and the elderly women above 40 years.