Co-existence of malnutrition and sarcopenia and its related factors in a long-term nursing care facility: A cross-sectional study.

Objectives: Malnutrition and sarcopenia often co-exist in older patients. This condition, called co-MS, shows a worse prognosis than either condition alone but is often overlooked and undertreated. We aimed to clarify the prevalence of co-MS and its associated factors with a focus on prescription in a long-term nursing care facility in Japan. Methods: Patients aged >65 years who resided in a long-term nursing care facility in Hyogo, Japan, were recruited for this cross-sectional study, which was conducted from July 1 to July 30, 2022. Sarcopenia and malnutrition were diagnosed using the Asian Working Group for Sarcopenia and Global Leadership Initiative on Malnutrition criteria, respectively. Patients who met both criteria were classified as having co-MS. Potentially associated factors, including age, sex, length of stay, activities of daily living, comorbidity, oral function and hygiene, swallowing ability, and the number and type of prescriptions, were assessed. Results: The prevalence of sarcopenia was 92 % (72/78). All malnourished patients were sarcopenic (40.3 %) and were classified as having co-MS. Oral function and hygiene, swallowing ability, comorbidity, and the presence of potentially inappropriate medications showed significant associations in univariate analyses. Of particular note, potentially inappropriate medication was an independent factor in the multivariate analysis. Conclusions: Co-MS is prevalent in long-term nursing care facilities; thus, healthcare workers should pay attention to relevant factors to identify patients at risk of co-MS and to provide appropriate care and intervention.

Self-reported seafood intake and atopy in Japanese school-aged children.

BACKGROUND: The effects of fish consumption and n-3 poly-unsaturated fatty acid (PUFA) levels on atopic disorders are inconsistent in previous reports, but few studies have investigated the effects of both fish and n-3 PUFA. The aim of the present study was to investigate whether erythrocyte fatty acids and the consumption of fish are associated with atopic diseases in pre- and early adolescents. METHODS: A total of 135 students with eczema, 136 students with asthma, and 137 healthy control students were selected from fifth and eighth grades in Shunan, Japan. Atopic disorders and dietary intake were evaluated with questionnaires, and total serum IgE was measured using an enzyme-linked immunosorbent assay. In addition, erythrocyte membrane levels of PUFA were assessed via gas chromatography. RESULTS: Total IgE was significantly elevated in the atopic subjects (P < 0.001). The intake of fatty and dried fish or seafood was significantly associated with eczema (odds ratios of the highest quartiles: 0.46, 95% confidence interval (95%CI): 0.22-0.94; 0.34, 95%CI: 0.16-0.71, respectively). Additionally, only erythrocyte eicosapentaenoic acid (EPA) level had a negative association with eczema (P= 0.048). For asthma, the effect of fish consumption was not significant. CONCLUSIONS: Fish consumption was related to a low prevalence of eczema, but not asthma in Japanese pre- and early adolescents. EPA may be involved in this mechanism.

Association of serum carotenoids and tocopherols with atopic diseases in Japanese children and adolescents.

The present study assessed whether serum carotenoids and tocopherols are associated with atopic diseases (eczema and asthma) in 10- and 13-yr-olds in a Japanese community. Of 2796 students attending schools in Shunan, Japan, in 2006, 396 students were randomly selected for this study using nested case-control design. Atopic diseases and dietary food intake were assessed using self-administered questionnaires, and serum antioxidants were analyzed using high-performance liquid chromatography. We found no associations between serum carotenoids and atopic diseases. However, odds ratios (OR)s for the third and fourth quartiles of serum alpha-tocopherol with atopic eczema were 0.33 (95% confidence interval: 0.15-0.73) and 0.36 (0.14-0.89), respectively, and the trend was negatively significant (P(trend) = 0.048). We did not find a significant association for asthma. In conclusion, serum alpha-tocopherol was negatively associated with the prevalence of eczema. Serum carotenoids did not show definitive protective effects in Japanese youth.

Occurrence of singlet oxygen oxygenation of oleic acid and linoleic acid in the skin of live mice.

To assess the contribution of singlet molecular oxygen [O(2) ((1)Delta(g))] to lipid peroxidation in vivo, this study combined gas chromatography-mass spectrometry with thin layer chromatography to analyse peroxidized lipids in the skin of hairless mice. Hydroxyoctadecenoate isomers and unconjugated hydroxyoctadecadienoate isomers derived from peroxidized oleic acid and linoleic acid, respectively, which are specific to O(2) ((1)Delta(g))-dependent oxygenation, were detected in the skin of live mice under ordinary feeding conditions. Short-term ultraviolet A (UVA)-irradiation of the skin in vivo elevated levels of the unconjugated hydroxyoctadecadienoate isomers significantly, whereas the irradiation of skin homogenate in vitro increased levels of all isomers derived from both O(2) ((1)Delta(g)) and free radical-dependent oxygenation to a much greater extent. This is the first report to demonstrate the occurrence of O(2) ((1)Delta(g))-specific oxygenation of unsaturated fatty acids in living animals.

Ingested beta-carotene enhances glutathione level and up-regulates the activity of cysteine cathepsin in murine splenocytes.

To elucidate health benefits of beta-carotene, especially on immunity, we measured redox-related indices in spleen cells from BALB/c mice supplemented with various amounts of beta-carotene. In mice supplemented with beta-carotene in their diet, glutathione, an intracellular anti-oxidation agent, increased in their splenocytes. This change was highly correlated with the accumulation of beta-carotene, but not with that of retinol. The increase in glutathione was accompanied by an increase in mRNA for gamma-glutamylcysteine synthetase, a rate-limiting enzyme for glutathione synthesis. The higher the glutathione content was in the spleen cells, the higher the activity of cysteine cathepsin became in crude antigen-presenting cells contained in the spleen. These data suggest that accumulated beta-carotene in splenocytes, without being metabolized, caused an increase in the intracellular glutathione level, thereby anti-oxidatively supporting the activity of redox-sensitive lysosomal protease, which is involved in antigen-presentation.

Combination of TLC blotting and gas chromatography-mass spectrometry for analysis of peroxidized cholesterol.

We have established a sensitive and convenient method for analysis of cholesterol hydroperoxides (Chol-OOHs) as trimethylsilyloxyl derivatives using diphenylpyrenylphosphine (DPPP)-thin-layer chromatography (TLC) blotting and gas chromatography-electron ionization-mass spectrometry/selected-ion monitoring (GC-EI-MS/SIM). Chol-OOH standards were prepared by photosensitized oxidation and azo radical-induced peroxidation of cholesterol. Trimethylsilyloxyl derivatives of cholesterol 5alpha-hydroperoxide (Chol 5alpha-OOH), cholesterol 7alpha-hydroperoxide (Chol 7alpha-OOH), and cholesterol 7beta-hydroperoxide (Chol 7beta-OOH) could be separated from one another in the SIM chromatogram using a fragment ion with elimination of trimethylsilanol from the molecular ion. This method was used to characterize peroxidized cholesterol from azo radical-exposed human low-density lipoprotein and UVA-irradiated human keratinocytes in the presence of hematoporphyrin. Finally, we succeeded in the quantification of each Chol-OOH isomer present in hairless mouse skin with and without UVA irradiation by use of beta-sitosterol hydroperoxide as internal standard. The accumulation of Chol 5alpha-OOH with Chol 7alpha/betaOOH in the skin indicates that singlet molecular oxygen ((1)O(2)) participated in the peroxidation of skin cholesterol, because Chol 5alpha-OOH is known to be a (1)O(2) specific cholesterol peroxidation product. We concluded that the combination of DPPP-TLC blotting and GC-EI-MS/SIM is useful for quantifying peroxidized cholesterol in biological samples and confirming the participation of (1)O(2) in oxidative stress.

Participation of singlet oxygen in ultraviolet-a-induced lipid peroxidation in mouse skin and its inhibition by dietary beta-carotene: an ex vivo study.

Dietary beta-carotene acts as a photoprotective agent in the skin, but the exact mechanism of protection is unknown. This ex vivo study is focused on determining the mechanism of action of beta-carotene against UV-A-induced skin damage by characterizing peroxidized phosphatidylcholine (PC) and beta-carotene oxidation products. BALB/c mice were fed with basal or a beta-carotene-supplemented diet, and homogenates from their dorsal skin were prepared after 3 weeks for UV-A irradiation. Analyses revealed that the degree of lipid peroxidation in the beta-carotene group was significantly lower than that in the controls. The isomeric composition of hydroperoxy fatty acids, constituting peroxidized PC, was determined by thin-layer chromatography-blotting followed by gas chromatography/mass spectrometry (MS)/selected ion monitoring analysis. The 9- and 10-isomers of peroxidized PC, resulting from the reaction of singlet molecular oxygen ((1)O(2)) with oleic acid, were elevated in the UV-A-exposed control group compared to the experimental group. Similar results were obtained from methylene-blue-sensitized photooxidation of mouse skin lipids in vitro. Liquid chromatography/MS analysis of the homogenates confirmed the formation of beta-carotene 5,8-endoperoxide, a specific marker for the (1)O(2) reaction. These results indicate that dietary beta-carotene accumulates in the skin and acts as a protective agent against UV-A-induced oxidative damage, by quenching the (1)O(2).

Alum augments the experimental allergenicity of Kunitz-type soybean trypsin inhibitor independent of the antigen-adsorption.

In order to inspect the significance of the adsorbing property in the adjuvant activity to enhance IgE production, we immunized BALB/c mice against Kunitz-type soybean trypsin inhibitor (KSTI), the most potent experimental allergen among soybean proteins, associated with Aluminum hydroxide (alum) or DEAE-Sephadex particles. The production of immunoglobulin isotypes was analyzed at the various amounts, 3-3,000 microg per mouse, of the antigen dosages. In our experiments, although alum did not adsorb KSTI significantly, it augmented the total and the antigen-specific IgE without affecting the optimal range of the antigen dosage. On the other hand, alum did not effectively enhance the production of the other immunoglobulin isotypes. The production of immunoglobulin isotypes other than IgE increased dose-dependently on the antigen. These results ensured our previous finding that another protein, ovalbumin, was used as the antigen. We also demonstrated that the adsorption of KSTI by DEAE-Sephadex in the immunizing vehicle resulted in the requirement of more KSTI for accomplishing the equal immunity in BALB/c mice compared to the control. Moreover, we demonstrated that, regardless of the inability to adsorb KSTI, alum exerted its adjuvant activity only when it was co-injected with the antigen. These results showed that some biochemical effect, other than adsorptive activity, to enhance the production of the antigen-specific IgE resides in alum.

Plasma metabolites of dietary flavonoids after combination meal consumption with onion and tofu in humans.

SCOPE: The effect of food combination on metabolic profile in postprandial plasma has hardly been reported. We investigated the absorption and metabolism of quercetin and soy isoflavones in humans after combination meal consumption. METHODS AND RESULTS: Five healthy volunteers ingested sautéed onion and tofu, and the plasma metabolites of quercetin and isoflavones were analyzed. Quercetin and genistein were incubated with human intestinal Caco-2 cells and human hepatoma HepG2 cells to further analyze the influence of simultaneous supply to the small intestine and the liver. Glucuronosyl conjugates of quercetin and methylated quercetin were the major plasma metabolites in the case of onion intake. Plasma metabolites with the single serving of tofu were both glucuronide and sulfate metabolites of isoflavones. Interestingly, quercetin sulfate was only detected after the combined intake of sautéed onion and tofu, accompanied with a decrease in sulfated isoflavones. Besides, quercetin was shown as the preferential substance for phase II enzymes over genistein in both Caco-2 and HepG2 cells. CONCLUSION: These results indicate that, when flavonoids and isoflavonoids were ingested together, the metabolic conversions in the small intestine and/or the liver could be altered, resulting in the variation of the postprandial profiles of the plasma metabolites.

Non-selective distribution of isomeric cholesterol hydroperoxides to microdomains in cell membranes and activation of matrix metalloproteinase activity in a model of dermal cells.

Cholesterol hydroperoxides (ChOOHs) are included as lipid peroxidation products in the skin exposed to ultraviolet (UV) light irradiation. They may exert physicochemical actions affecting biomembrane rigidity because cholesterol is one of the major components of cell membranes. We investigated the distribution of isomeric ChOOHs in heterogeneous cell membranes with different lipid profiles using mouse fibroblast NIH-3T3 cells as a model of the dermis. Before and after UVA irradiation in the presence of hematoporphyrin, cell membranes were partitioned to microdomains (lipid rafts and caveolae) containing a higher amount of cholesterol and non-microdomains (containing a lower amount of cholesterol) by ultracentrifugation. By a combination of diphenylpyrenylphosphine-thin-layer chromatography blotting analyses and gas chromatography-electron ionization-mass spectrometry/selected ion monitoring analyses, ChOOH isomers were determined as their trimethylsilyloxyl derivatives. Cholesterol 5α-, 7α- and 7ß-hydroperoxide were found as isomeric ChOOHs before irradiation. The amounts of the three ChOOH isomers increased significantly after photoirradiation for 2h. No difference was observed between microdomains and non-microdomains with regard to the ratio of the amounts of isomeric ChOOHs to that of cholesterol, suggesting that these ChOOH isomers were distributed equally in both parts depending on cholesterol content. When cells were treated with a purified mixture of ChOOH isomers, cell membranes incorporated ChOOHs into microdomains as well as non-microdomains evenly. Cellular matrix metalloproteinase-9 (MMP-9) activity was elevated by treatment with the purified mixture of ChOOH isomers. These results strongly suggest that ChOOHs accumulate in cell membranes irrespective of the heterogeneous microstructure and promote MMP activity if dermal cells are exposed to photodynamic actions.

Singlet molecular oxygen-quenching activity of carotenoids: relevance to protection of the skin from photoaging.

Carotenoids are known to be potent quenchers of singlet molecular oxygen [O(2) ((1)Δ(g))]. Solar light-induced photooxidative stress causes skin photoaging by accelerating the generation of reactive oxygen species via photodynamic actions in which O(2) ((1)Δ(g)) can be generated by energy transfer from excited sensitizers. Thus, dietary carotenoids seem to participate in the prevention of photooxidative stress by accumulating as antioxidants in the skin. An in vivo study using hairless mice clarified that a O(2) ((1)Δ(g)) oxygenation-specific peroxidation product of cholesterol, cholesterol 5α-hydroperoxide, accumulates in skin lipids due to ultraviolet-A exposure. Matrix metalloproteinase-9, a metalloproteinase family enzyme responsible for the formation of wrinkles and sagging, was enhanced in the skin of ultraviolet-A -irradiated hairless mice. The activation of metalloproteinase-9 and the accumulation of 5α-hydroperoxide, as well as formation of wrinkles and sagging, were lowered in mice fed a ß-carotene diet. These results strongly suggest that dietary ß-carotene prevents the expression of metalloproteinase-9 (at least in part), by inhibiting the photodynamic action involving the formation of 5α-hydroperoxide in the skin. Intake of ß-Carotene therefore appears to be helpful in slowing down ultraviolet-A -induced photoaging in human skin by acting as a O(2) ((1)Δ(g)) quencher.

Ingested quercetin but not rutin increases accumulation of hepatic beta-carotene in BALB/c mice.

Beta-carotene is a carotenoid with a range of reported health benefits besides vitamin A activity. If the enzymatic conversion of beta-carotene to retinal is suppressed in the digestive tract, residual beta-carotene that reaches the tissues increases. We evaluated the function of quercetin and rutin (quercetin-3-rutinoside) to increase the accumulation of beta-carotene in vitro and in vivo in BALB/c mice. When the conversion of beta-carotene by a preparation of the murine small intestine was measured in vitro, the addition of quercetin or rutin considerably inhibited the conversion. When the levels of hepatic beta-carotene and retinoids were measured among three groups of mice fed a diet supplemented with beta-carotene plus quercetin or rutin or beta-carotene alone (four to six mice per group), quercetin increased the level of beta-carotene and decreased the level of retinol, whereas rutin did not. These results demonstrate that quercetin can suppress the conversion of beta-carotene which develops in the cytosol of small intestinal epithelial cells, and that rutin whose rutinose-moiety prevents being absorbed in the small intestine cannot suppress the conversion in vivo. This study offers a novel insight into the interaction between flavonoids and carotenoids with respect to the health benefits from the latter.

beta-Carotene and beta-cryptoxanthin but not lutein evoke redox and immune changes in RAW264 murine macrophages.

The mechanism of immunological benefits induced by carotenoids has not been fully elucidated. Here, we investigated some of the immunity-related properties of beta-carotene and two other carotenoids, beta-cryptoxanthin, and lutein, on the murine macrophages cell line RAW264. beta-Carotene added to the culture medium accumulated in the cells in a time- and dose-dependent manner. The accumulation was positively correlated with cellular lipid peroxidation, demonstrating the pro-oxidative activity of beta-carotene, and also with the synthesis of glutathione, an intracellular antioxidant. Conversely, accumulation of beta-carotene was negatively correlated with the transcription of immune-active molecules, such as IL-1beta, IL-6, and IL-12 p40, in cells stimulated by LPS and INF-gamma. The transcription of the pro-inflammatory cytokines IL-1beta and IL-6 was more sensitive to the accumulation of beta-carotene than was IL-12 p40. The accumulation of beta-cryptoxanthin in cells resulted in effects similar to those of beta-carotene. However, lutein accumulated minimally and did not significantly affect the cells. These results demonstrate that beta-carotene, and beta-cryptoxanthin as well, can accumulate in RAW264 cells and induce changes in intracellular redox status, which in turn regulate the immune function of macrophages.

Peroxidized cholesterol-induced matrix metalloproteinase-9 activation and its suppression by dietary beta-carotene in photoaging of hairless mouse skin.

The activation of matrix metalloproteinase (MMP)-9 leading to the formation of wrinkle and sagging of skin is an essential step in the skin photoaging on exposure to ultraviolet A (UVA). This study attempted to elucidate the role of peroxidized cholesterol including cholesterol hydroperoxides (Chol-OOHs), primary products of lipid peroxidation in biomembranes, in MMP-9 activation and the effect of dietary beta-carotene in MMP-9 activation. Hairless mice were subjected to periodic UVA irradiation for 8 weeks. The amount of peroxidized cholesterol detected as total hydroxycholesterol in the skin was increased significantly by the exposure. The activity and protein level of MMP-9 were elevated with wrinkling and sagging formation. MMP-9 activity was also enhanced by the intracutaneous injection of Chol-OOHs into the mouse skin. Adding beta-carotene to the diet of the mice during the period of irradiation suppressed the activity and expression of MMP-9 as well as the wrinkling and sagging formation. The amount of cholesterol 5alpha-hydroperoxide, a singlet molecular oxygen oxygenation-specific peroxidized cholesterol, was significantly lowered by the addition of beta-carotene to the diet. These results strongly suggest that Chol-OOHs formed on exposure to UVA contribute to the expression of MMP-9, resulting in photoaging. Dietary beta-carotene prevents the expression of MMP-9, at least partly, by inhibiting photodynamic action involved in the formation of Chol-OOHs.

Carotenoid, tocopherol, and fatty acid biomarkers and dietary intake estimated by using a brief self-administered diet history questionnaire for older Japanese children and adolescents.

We investigated the association between nutrient biomarkers and dietary intake estimated using a brief self-administered dietary history questionnaire (BDHQ) for Japanese children and adolescents. Blood samples were collected from 398 subjects (5th graders of elementary school aged 10-11 y, and 2nd graders of secondary schools aged 13-14 y) randomly selected from among students in Shunan City, Japan, who were then required to answer two questionnaires. Spearman correlations were calculated between dietary intake and the corresponding biomarkers (serum carotenoids, tocopherols, and erythrocyte fatty acids). Correlations with beta-carotene and beta-cryptoxanthin were significant in the 13- and 14-y age group (r=0.220-0.333, p<0.030) and the 10- and 11-y age subgroup who answered the questionnaire with assistance (r=0.295-0.299, respectively, p=0.006). Consumption of green-yellow vegetables and fruits was significantly correlated with beta-carotene and beta-cryptoxanthin levels (r=0.205-0.341, p<0.047). In the 13- and 14-y age group, correlations with eicosapentaenoic and docosahexaenoic acids were between 0.215 and 0.473 (p<0.040). Total seafood intake was significantly correlated with marine n-3 polyunsaturated fatty acids (PUFAs; r=0.239-0.420, p<0.023). In the 10- and 11-y age subgroup who completed the questionnaire with assistance, seafood intake was significantly correlated with marine n-3 PUFAs (r=0.239-0.243, p<0.032). In conclusion, dietary intake assessed using the BDHQ reflects the corresponding biomarkers for 13- and 14-y-olds; however, when used for elementary school children, caution is necessary in interpreting the results.

Effect of an excessive intake of quercetin on the vitamin E level and antioxidative enzyme activities of mouse liver under paraquat-induced oxidative stress.

The liver alpha-tocopherol level of the paraquat fed mice group was lower than that of the control diet-fed group. An excessive intake of quercetin lowered the liver alpha-tocopherol level of the control diet-fed mice group, but did not affect it in the paraquat-fed mice group. The same quercetin intake significantly increased the superoxide dismutase and glutathione peroxidase activities in the liver of both groups, indicating that excessive quercetin intake can either promote or attenuate oxidative stress in the liver.

Beta-carotene modulates the immunological function of RAW264, a murine macrophage cell line, by enhancing the level of intracellular glutathione.

The activities of beta-carotene on redox status and the immune functions of RAW264 cells, a murine macrophage cell line, were investigated. Supplementation with beta-carotene for RAW264 cells resulted in apparently inconsistent redox indices: lipid peroxidation was enhanced but intracellular oxidation was moderately attenuated. Attenuated intracellular oxidation was endorsed by an increase in glutathione accompanied by up-regulated transcription of a subunit of gamma-glutamylcysteine synthetase, the rate-limiting enzyme for glutathione synthesis. alpha-Tocopherol, which can quench lipid peroxidation by free radical, neither inhibited that by beta-carotene nor influenced the intracellular redox status. Lipopolysaccharide-stimulated transcriptions of IL-1beta and IL-12 p40 in RAW264 were inhibited by beta-carotene but not by alpha-tocopherol. These results indicate that beta-carotene, which can modulate the intracellular redox status of macrophages by enhancing the level of intracellular glutathione, is related to the immune functions of macrophages.

Feeding with both ß-carotene and supplemental α-tocopherol enhances type 1 helper T cell activity among splenocytes isolated from DO11.10 mice.

beta-Carotene and/or supplemental alpha-tocopherol were fed to DO11.10 mice to investigate their effect on the immune function of naive splenocytes. A high secretion of interleukin-12 and interferon-gamma in response to the ex vivo primary antigen presentation occurred only when both were fed. This is consistent with the suppressed immunoglobulin E production under the similar condition described in our previous report.

Inhibition of immunoglobulin E production in allergic model mice by supplementation with vitamin E and beta-carotene.

A diet containing different amounts of vitamin E (alpha-tocopherol; 0.5 mg, 5 mg, 10 mg or 50 mg per 100 g diet) was supplemented to BALB/c mice for 6 weeks. These mice were subcutaneously immunized twice with ovalbumin (OVA). A passive cutaneous anaphylaxis (PCA) analysis demonstrated that the mice fed on the diet containing 5 mg of vitamin E produced the highest level of the OVA-specific immunoglobulin E (IgE) antibody. A lower level of serum IgE was found in the mice supplemented with 0.5 mg, 10 mg and 50 mg of vitamin E. A sandwich ELISA analysis showed that the pattern of the total IgE antibody level among these four groups was the same as that of the allergen-specific IgE. In a separate experiment, 5 mg of vitamin E and/or 50 mg of beta-carotene was supplemented to the basal diet containing vitamin E as alpha-tocopherol acetate (5 mg) in order to evaluate the effect of their combination on OVA-specific and total IgE production in the mice. The supplementation with beta-carotene alone had no effect on OVA-specific or total IgE production. In contrast, supplementation with vitamin E plus beta-carotene effectively suppressed both the antigen-specific and total IgE antibodies. The serum vitamin E and beta-carotene levels were increased by supplementation with the respective compounds. These results strongly suggest that the combination of dietary vitamin E and beta-carotene suppressed IgE production and would therefore help to prevent the type-I allergic reaction.