Effect of Tempeh and Daidzein on Calcium Status, Calcium Transporters, and Bone Metabolism Biomarkers in Ovariectomized Rats.

Menopause marks a critical life stage characterized by hormonal changes that significantly impact bone health, leading to a heightened susceptibility to bone fractures. This research seeks to elucidate the impact of daidzein and tempeh on calcium status, calcium transporters, and bone metabolism in an ovariectomized rat model. Forty female Wistar rats, aged 3 months, participated in a two-phase experiment. The initial phase involved inducing a calcium deficit, while the second phase comprised dietary interventions across five groups: Sham (S) and Ovariectomy (O) with a standard diet, O with bisphosphonate (OB), O with pure daidzein (OD), and O with tempeh (OT). Multiple parameters, encompassing calcium levels, calcium transporters, bone histopathology, and serum bone metabolism markers, were evaluated. The findings revealed that the OT group showcased heightened levels of bone turnover markers, such as pyridinoline, C-telopeptide of type I collagen, bone alkaline phosphatase, and procollagen type I N-terminal propeptide, in contrast to S and O groups, with statistical significance (p < 0.05). Histopathologically, both the OD and OT groups exhibited effects akin to the OB group, indicating a decrease in the surface area occupied by adipocytes in the femoral bone structure, although statistically non-equivalent, supporting the directionally similar trends. Although TRPV5 and TRPV6 mRNA expression levels in the jejunum and duodenum did not display statistically significant differences (p > 0.05), the OD and OT groups exhibited increased expression compared to the O group. We hypothesized that obtained results may be related to the effect of isoflavones on estrogen pathways because of their structurally similar to endogenous estrogen and weak estrogenic properties. In conclusion, the daily consumption of pure daidzein and tempeh could potentially improve and reinstate calcium status, calcium transport, and bone metabolism in ovariectomized rats. Additionally, isoflavone products demonstrate effects similar to bisphosphonate drugs on these parameters in ovariectomized rats.

Influence of Isoflavones and Probiotics on Magnesium Status in Healthy Female Rats.

Isoflavones and probiotics are promising nutrients for bone health, and magnesium (Mg) is essential for bone metabolism. This study aimed to determine the effects of daidzein, genistein and Lactobacillus acidophilus on the Mg status of healthy female rats. Forty-eight rats were randomly assigned to six groups, with the control group receiving a standard diet (AIN 93M). The remaining groups were fed the same diet with added ingredients such as tempeh flour; soy flour; pure daidzein and genistein; L. acidophilus or a combination of daidzein, genistein, and L. acidophilus. Tissue samples were collected after the eight-week intervention, and Mg concentrations were analysed using flame atomic absorption spectrometry. Myeloid and erythroid cells were determined using the haematoxylin and eosin bone staining method. Statistical analyses were conducted using one-way ANOVA with Tukey's post hoc test and Pearson's correlation coefficient. The threshold for significance was p < 0.05. Compared with the control group, adding tempeh to the diet of rats resulted in significant changes in Mg concentrations in various tissues, with a decrease in the kidneys and an increase in the fur. Although not statistically significant compared to the control group, the tempeh group showed increased Mg concentrations in the femur and spleen. The myeloid-to-erythroid cell ratio did not differ significantly among groups, but all intervention groups showed higher ratios than the control group. A strong negative correlation was observed between Mg concentrations in the kidneys and fur. Conversely, a positive correlation was identified between Mg concentrations in the pancreas and fur. Daily consumption of tempeh may improve Mg status in the organism. Intake of pure daidzein, genistein, or probiotic seems to have no effect on Mg concentrations in healthy rats.

Effects of isoflavone and probiotic intake on calcium transport and bone metabolism biomarkers in female rats.

Calcium is essential for maintaining bone health as it contributes to bone formation, remodeling, strength, and density. This study investigated the effect of isoflavones and probiotics on calcium transporters' gene expression, serum calcium levels, and bone metabolism biomarkers in healthy female rats. Forty-eight female Wistar rats were classified into six groups. Bone metabolism biomarkers (pyridinoline, deoxypyridinoline, parathyroid hormone, and osteocalcin) and serum calcium levels were measured by enzyme-linked immunosorbent assay (ELISA) and atomic absorption spectroscopy (AAS), respectively. Gene expression of calcium transporters (Trpv5 and Trpv6) was evaluated in duodenum and jejunum tissue samples using quantitative polymerase chain reaction (qPCR). Trpv5 and Trpv6, epithelial calcium channels, play a crucial role in calcium transport and homeostasis in the body. The study consisted of a1-week adaptation period for the rats to adjust to the controlled conditions, followed by an 8-week intervention phase. The daidzein and genistein group showed a significant increase in the gene expression of the Trpv6 transporter in the duodenum and a marked decrease in serum pyridinoline levels compared to the control group. The tempeh and soybean groups showed a significant decrease in the gene expression of the Trpv5 calcium transporter in the jejunum. However, no significant influence of the Lactobacillus acidophilus diet on calcium transport and bone metabolism biomarkers was observed in the L. acidophilus group. The correlation analysis showed a significant positive relationship between serum calcium, bone metabolism biomarkers, and calcium transporters. In conclusion, our study demonstrates that the daidzein and genistein diet improves calcium transport in the duodenum and reduces pyridinoline serum concentrations, while tempeh and soybean diets reduce calcium transport in the jejunum. However, the combination of daidzein, genistein, and L. acidophilus did not demonstrate a synergistic effect on calcium transport and bone metabolism, suggesting that further investigations are needed to elucidate their potential interactions.

Corrigendum to "Isoflavones and probiotics effect on bone calcium and bone cells in rats" [Heliyon 9(6) (June 2023) e16801].

[This corrects the article DOI: 10.1016/j.heliyon.2023.e16801.].

Isoflavones and probiotics effect on bone calcium and bone cells in rats.

Isoflavones and probiotics have shown the therapeutic potential to alter calcium absorption and bone cell metabolism. This study sought to ascertain the effect of isoflavones and probiotics on calcium status and bone health in healthy female rats. Forty-eight adult female Wistar rats were grouped and fed: a standard diet (control); and standard diets with tempeh; soy; daidzein and genistein; Lactobacillus acidophilus; and a combination of daidzein, genistein, and L. acidophilus. The biochemical serum parameters, such as alanine transaminase, aspartate transaminase, glucose, and triacylglycerol concentrations, were measured, and calcium contents in tissues were determined. After staining the bone with hematoxylin and eosin, the number of osteoblasts, osteocytes, and the percentage of bone marrow adipocytes were counted. Compared with the control group, the soy group showed a significantly lower triacylglycerol concentration. The L. acidophilus group considerably increased the calcium content in the femoral bone. The daidzein and genistein, L. acidophilus, and a combination of daidzein, genistein, and L. acidophilus groups showed significantly lower calcium contents in the heart and kidneys. The daidzein and genistein group significantly enhanced the number of osteoblasts and osteocytes. A substantial inverse correlation was observed between calcium contents in kidneys and osteoblasts. In conclusion, the combination of daidzein, genistein, and L. acidophilus may improve bone calcium concentrations and bone cells. However, no synergistic effect between isoflavones and probiotics was detected in this study.

The impact of soy products, isoflavones, and Lactobacillus acidophilus on iron status and morphological parameters in healthy female rats.

BACKGROUND: Isoflavones and probiotics are two major factors involved in bone health. Osteoporosis and disturbances in iron (Fe) levels are common health problems in aging women. This study aimed to analyze how soybean products, daidzein, genistein, and Lactobacillus acidophilus (LA) affect Fe status and blood morphological parameters in healthy female rats. METHODS: A total of 48 Wistar rats aged 3 months were randomly divided into six groups. The control group (K) received a standard diet (AIN 93 M). The remaining five groups received a standard diet supplemented with the following: tempeh flour (TP); soy flour (RS); daidzein and genistein (DG); Lactobacillus acidophilus DSM20079 (LA); as well as a combination of daidzein, genistein, and L. acidophilus DSM20079 (DGLA). After 8 weeks of intervention, blood samples of the rats were collected for morphological analysis, whereas tissue samples were collected and kept at -80 °C until Fe analysis. Red blood cells, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets (PLTs), red cell distribution width, white blood cells, neutrophils (NEUT), lymphocytes (LYM), monocytes, eosinophils (EOS), and basophils were measured for blood morphological analysis. Fe concentrations were determined using flame atomic spectrometry. For statistical analysis, an ANOVA test for significance at the 5 % level was used. The relationship between tissue Fe levels and blood morphological parameters was determined using Pearson's correlation. RESULTS: Although no significant differences were observed in the Fe content between all diets, the TP group showed significantly higher levels of NEUT and lower levels of LYM than the control group. Compared with the DG and DGLA groups, the TP group showed a dramatically higher PLT level. In addition, the RS group showed significantly higher Fe concentrations in the spleen compared with the standard diet. Compared with the DG, LA, and DGLA groups, the RS group also showed significantly higher Fe concentrations in the liver. Compared with the TP, DG, LA, and DGLA groups, the RS group showed dramatically higher Fe concentrations in the femur. Pearson's correlations between blood morphological parameters and Fe levels in tissues were observed, especially a negative correlation between the Fe level in the femur and the NEUT concentration (-0.465) and a strong positive correlation between the Fe level in the femur and the LYM concentration (0.533). CONCLUSION: Soybean flour was found to increase Fe levels in rats, whereas tempeh may alter anti-inflammatory blood parameters. Isoflavones and probiotics did not affect Fe status in healthy female rats.

Carob kibbles as an alternative raw material for production of kvass with probiotic potential.

BACKGROUND: Non-diary beverages with probiotic properties are of great interest nowadays. In this research, we evaluated the suitability of carob kibbles in the manufacture of kvass. Kvass is a low-alcohol drink popular in Central and Eastern Europe and indicated as a potential non-diary beverage with probiotic properties. Therefore, the viability of probiotic strains of Lactobacillus plantarum and Saccharomyces boulardii during 4 weeks' storage in manufactured beverages was tested. RESULTS: Carob kibbles introduced significant amounts of phenolic compounds into kvasses, especially gallic acid (up to 117.45 ± 10.56 mg L-1 ), and improved antiradical activity up to 78% after fermentation. Moreover, fermentation efficiently reduced furfural and hydroxymethylfurfural content in samples up to 12.9% and 29.9%, respectively. Kvasses with rye malt extract possessed coffee-like, chocolate-like, roasted and caramel-like odours and a more bitter taste. Whereas kvass with carob kibbles was characterized by fruit-like odour and sweeter taste. Fermentation contributed to a creation as well as degradation of volatiles. L. plantarum exhibited higher general mortality during storage, whereas, in the case of S. boulardii, the viability was significantly higher regardless of the sample composition. CONCLUSION: This is the first study reporting the use of carob kibbles for kvass production. The obtained results showed that carob kibbles can replace rye malt extract, at least partially, in the production of kvass, giving to the product added health benefits. Moreover, S. boulardii is a better choice for production of kvass with probiotic properties. © 2021 Society of Chemical Industry.

Influence of fermentation by different microflora consortia on pulque and pulque bread properties.

BACKGROUND: Pulque bread is a traditional Mexican product obtained by fermentation using microflora present only in pulque. In this study, the possibility of creating a pulque microbial consortium under laboratory conditions and its applications were evaluated. A laboratory-made consortium was compared with a consortium originating in Mexico in bread and pulque production. They were tested in various growth medium systems: pulque made from agave sap and malt extract, Mexican wheat and rye pulque bread, and European wheat and rye bread. RESULTS: Depending on the growth medium, consortiums showed differing influence on many factors, such as specific volume, weight loss after baking, soluble proteins, and crust and crumb color. Indigenous starters increased sensorial acceptance of pulque and Mexican rye bread, decreased pH, and increased titratable acidity of the breads at the highest level whereas laboratory consortia improved sensory acceptance of wheat breads. The laboratory-prepared starter in some cases improved antiradical activity. All pulques received similar consumer evaluations. However, malt pulque was the least appreciated beverage. CONCLUSION: The results show the possibility of creating a pulque microbial consortium under laboratory conditions. Depending on the flour type and the breadmaking technique, the use of a particular microbial consortium allowed modification of certain physicochemical parameters. In conclusion, it is feasible to modify bread parameters to obtain features corresponding to consumer demands by using an appropriate microflora, pulque, or flour type. Moreover, this research describes, for the first time, the use of rye malt for pulque and rye flour for pulque bread preparation as raw materials. © 2019 Society of Chemical Industry.

Development of a Sustainable, Simple, and Robust Method for Efficient l-DOPA Extraction.

l-3,4-dihydroxyphenylalanine (l-DOPA) is a medically relevant compound in Parkinson's disease therapy. Several extraction methods of l-DOPA from beans, including velvet and faba beans, have been described in the literature. However, these methods require the use of strong acids, long extraction times, or complex downstream processing, which makes the extraction of l-DOPA expensive and energy-demanding, limiting its industrial application. In addition, the stability of l-DOPA during the extraction process is critical, further complicating the extraction of adequate amounts of this amino acid. This work is the first report on a simple, rapid, greener, and robust extraction method of l-DOPA. The developed method consists of a quick homogenization step followed by a double extraction with 0.2% v/v acetic acid for 20 min and was applied to faba bean at a ratio of 1:25 with respect to the extracting solvent. This study also investigated the stability of l-DOPA during extraction and thermal treatment. The proposed method demonstrated to be robust and extraordinarily efficient for numerous cultivars of faba bean, velvet bean, and food products containing faba beans.

Effect of Replacing Cocoa Powder by Carob Powder in the Muffins on Sensory and Physicochemical Properties.

The increasing demand for cocoa and search for ingredients rich in bioactive compounds encouraged us to investigate the possibility of replacing it by carob powder in the muffins containing soy beans, sesame oil and flaxseeds. There was 5% addition of carob or cocoa powder to the individual doughs. The muffins with the addition of carob were characterized by improved antiradical activity (by 36% - 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) method, by 83% - 2,2-diphenyl-1-picrylhydrazyl (DPPH) method), higher content of genistein (18%) and total phytosterols (17%) in the dry mass. The color differences in the carob muffins crusts were not perceptible by consumers (ΔE = 0.70 for crust, ΔE = 5.6 for crumb) and their taste was found to be less bitter and sweeter than the taste of cocoa muffins. Moreover, the addition of carob powder as well as cocoa powder resulted in good sensory quality. The high content of phytosterols, genistein and improved antiradical properties proved carob to be a source of bioactive compounds. The results show that carob powder may be used as valuable alternative muffin ingredient to cocoa.

Evaluation of bean and soy tempeh influence on intestinal bacteria and estimation of antibacterial properties of bean tempeh.

In this study the effect of bean tempeh on the growth of Bacillus subtilis, Escherichia coli, Lactobacillus acidophilus and Lactobacillus paracasei bacteria was investigated. Antibacterial activity was observed only in relation to the bacteria Bacillus subtilis. The effect of tempeh products on human intestinal microflora was also assessed. Bean and soy tempeh were culinarily processed and next digested in conditions simulating the human digestive tract (one of the digestive tracts was equipped with a mechanism simulating absorption). Soy tempeh stimulated most the growth of bacteria of the genus Bifidobacterium, while bean tempeh that of Escherichia coli. Using simulation of absorption for the digestion of fried soy tempeh resulted in a higher rise in the bacteria count of the genus Lactobacillus, while after digestion of fried bean tempeh the highest increase was recorded for Bifidobacterium and E. coli.

Determination of compounds responsible for tempeh aroma.

Tempeh is a fermented food, popular mainly in south-east Asia, but also among vegetarians worldwide. It is produced by fermenting soybean or other beans with Rhizopus strains and usually eaten deep-fried, steamed or roasted. The flavour of tempeh depends upon the fermentation time, beans used and the (eventual) frying process. Our goal was to identify compounds responsible for the unique aroma of fermented and fried soy tempeh. Gas chromatography-olfactometry (GC-O) with the aroma extract dilution analysis (AEDA) approach, was used to determine key odorants after 1 and 5 days of fermentation and subsequent frying. Comprehensive gas chromatography-mass spectrometry (GC×GC-ToF-MS) was used for their quantitation using stable isotope dilution analysis (SIDA) or standard addition (SA) methods. Odour activity values (OAV) were calculated for 19 out of 21 key odorants. Tempeh was fermented for 5 days and fried, and the main aroma compounds were found to be the following: 2-acetyl-1-pyrroline, (FD=1024, OAV 1380), 2-ethyl-3,5-dimethylpyrazine (FD=512, OAV 338), dimethyl trisulfide, (FD=512, OAV 900), methional (FD=512, OAV 930), 2-methylpropanal (FD=512, OAV 311) and (E,E)-2,4-decadienal (FD=512, OAV 455). The frying process induced the increase or appearance of the main key odorants in tempeh.