Consumption of 2 Green Kiwifruits Daily Improves Constipation and Abdominal Comfort-Results of an International Multicenter Randomized Controlled Trial.

INTRODUCTION: Consumption of green kiwifruit is known to relieve constipation. Previous studies have also reported improvements in gastrointestinal (GI) comfort. We investigated the effect of consuming green kiwifruit on GI function and comfort. METHODS: Participants included healthy controls (n = 63), patients with functional constipation (FC, n = 60), and patients with constipation-predominant irritable bowel syndrome (IBS-C, n = 61) randomly assigned to consume 2 green kiwifruits or psyllium (7.5 g) per day for 4 weeks, followed by a 4-week washout, and then the other treatment for 4 weeks. The primary outcome was the number of complete spontaneous bowel movements (CSBM) per week. Secondary outcomes included GI comfort which was measured using the GI symptom rating scale, a validated instrument. Data (intent-to-treat) were analyzed as difference from baseline using repeated measures analysis of variance suitable for AB/BA crossover design. RESULTS: Consumption of green kiwifruit was associated with a clinically relevant increase of ≥ 1.5 CSBM per week (FC; 1.53, P < 0.0001, IBS-C; 1.73, P = 0.0003) and significantly improved measures of GI comfort (GI symptom rating scale total score) in constipated participants (FC, P < 0.0001; IBS-C, P < 0.0001). No significant adverse events were observed. DISCUSSION: This study provides original evidence that the consumption of a fresh whole fruit has demonstrated clinically relevant increases in CSBM and improved measures of GI comfort in constipated populations. Green kiwifruits are a suitable dietary treatment for relief of constipation and associated GI comfort.

Inflammatory markers and bone health in postmenopausal women: a cross-sectional overview.

BACKGROUND: Cytokines, chemokines, C-reactive proteins (CRP) and ferritin are known inflammatory markers. However, cytokines such as interleukin (IL-1ß), (IL-6) and tumour necrosis factor (TNF-α) have been reported to interfere with both the bone resorption and bone formation processes. Similarly, immune cell cytokines are known to contribute to inflammation of the adipose tissue especially with obesity. IL-10 but not IL-33 has been linked to lower ferritin levels and anemia. In this study, we hypothesized that specific cytokine levels in the plasma of women with low bone mineral density (BMD) would be higher than those in the plasma of healthy women due to the actions of elevated levels of pro-inflammatory cytokines in inducing osteoclast formation and differentiation during senescence. RESULTS: Levels of cytokines (IFNα2, IFN-γ, IL-12p70, IL-33) and monocyte chemoattractant protein-1 (MCP-1) were significantly higher in the plasma of the osteoporotic group compared to the osteopenic and/or healthy groups. Meanwhile CRP levels were significantly lower in women with osteoporosis (P = 0.040) than the osteopenic and healthy groups. Hip BMD values were significantly lower in women with high/detectable values of IL-1ß (P = 0.020) and IL-6 (P = 0.030) compared to women where these were not detected. Similarly, women with high/detectable values of IL-1ß had significantly lower spine BMD than those where IL-1ß was not detected (P = 0.030). Participants' CRP levels were significantly positively correlated with BMI, fat mass and fat percentage (P < 0.001). In addition, ferritin levels of women with high/detectable values of anti-osteoclastogenic IL-10 (P = 0.012) and IL-33 (P = 0.017) were significantly lower than those where these were not detected. There was no statistically significant association between TNF-α and BMD of the hip and lumbar spine. CONCLUSIONS: High levels of cytokines (IFNα2, IFN-γ, IL-12p70, IL-33) and MCP-1 in apparently healthy postmenopausal women are associated with bone health issues. In addition, an increase in levels of IL-10 and IL-33 may be associated with low ferritin levels in this age group. TRIAL REGISTRATION: ANZCTR, ACTRN12617000802303. Registered May 31st, 2017, https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=373020.

Calcium and vitamin D fortified milk reduces bone turnover and improves bone density in postmenopausal women over 1 year.

PURPOSE: In Malaysia, hip fracture incidence is higher in Chinese women than other ethnic groups. This study compared the effects of a high-calcium vitamin D fortified milk with added FOS-inulin versus regular milk over 1 year on aspects of bone health in Chinese postmenopausal women in Malaysia. METHODS: One-hundred and twenty-one women (mean age 59 (± 4) years) were randomized into two groups: control (n = 60; regular milk, 428 mg calcium per day) or intervention (n = 61; fortified milk at 1200 mg calcium, 96 mg magnesium, 2.4 mg zinc, 15 µg vitamin D and 4 g FOS-inulin per day). At baseline, weeks 12, 24, 36 and 52, parathyroid hormone (PTH), C-Telopeptide of Type I Collagen (CTx-1), Procollagen I Intact N-Terminal propeptide (PINP) and vitamin D levels were assessed. Bone density (BMD) was measured at baseline and week 52 using a GE Lunar iDXA. RESULTS: Body mass index, lumbar spine and femoral neck BMD did not differ between groups at baseline. Over 52 weeks, mean plasma 25 (OH) D3 levels increased to 74.8 nmol/L (intervention group) or remained at 63.1 nmol/L (control group) (p < 0.001 between groups). PTH levels increased in the control group (p = 0.001). The intervention resulted in a significant suppression of CTx-1 and PINP at p = 0.018 and p = 0.004. Femoral neck BMD remained stable in the intervention group but decreased significantly in the controls, with a borderline treatment effect (p = 0.07). CONCLUSION: Compared with regular milk, the fortified milk suppressed bone turnover markers and tended to increase femoral neck BMD.

Differential effects of calcium- and vitamin D-fortified milk with FOS-inulin compared to regular milk, on bone biomarkers in Chinese pre- and postmenopausal women.

PURPOSE: To compare the effects of a high-calcium vitamin D-fortified milk with added FOS-inulin versus regular milk on serum parathyroid hormone, and bone turnover markers in premenopausal (Pre-M) and postmenopausal (PM) women over 12 weeks. METHODS: Premenopausal women (n = 136, mean age 41 (±5) years) and postmenopausal women [n = 121, mean age 59 (±4) years] were recruited, and each age group randomised into two groups to take two glasses per day of control = regular milk (500 mg calcium per day) or intervention (Int) = fortified milk (1000 mg calcium for pre-M women and 1200 mg calcium for PM women, 96 mg magnesium, 2.4 mg zinc, 15 µg vitamin D, 4 g FOS-inulin per day). At baseline, week 4 and week 12 serum minerals and bone biochemical markers were measured and bone density was measured at baseline. RESULTS: Mean 25-hydroxyvitamin D [25(OH) vitamin D3] levels among groups were between 49 and 65 nmol/L at baseline, and over the 12 weeks of supplementation, the fortified milk improved vitamin D status in both Int groups. CTx-1 and PINP reduced significantly in both Pre-M and PM groups over the 12 weeks, with the changes in CTx-1 being significantly different (P < 0.035) between PM control and PM Int groups at week 12. Parathyroid hormone levels were significantly reduced in all groups over time, except for control PM group where levels increased at 12 weeks. CONCLUSION: The overall pattern of responses indicates that while both regular milk and fortified milk reduce bone resorption in young and older women, fortified milk is measurably more effective.

Bovine complex milk lipid containing gangliosides for prevention of rotavirus infection and diarrhoea in northern Indian infants.

Rotavirus (RV) is a leading cause of morbidity and mortality in children younger than 5 years of age, presenting commonly with diarrhoeal symptoms. In a prospective 12-week double-blind randomised controlled trial we assessed acceptability and efficacy of a high-ganglioside complex milk lipid (CML) for prevention of RV infection in 450 infants, ages 8 to 24 months, at 3 sites in northern India. Prevalence of diarrhoea and RV was unseasonably low at baseline (all-cause diarrhoea [ACD], n = 16; RV diarrhoea [RVD], n = 2; RV infection, RV positive [RV+], n = 20) and throughout the trial, with only 110 total episodes of ACD for 12 weeks (CML, n = 62; control, n = 48) of which 10 were RVD (CML, n = 4; control, n = 6). Mean duration that RVD persisted was lower in the CML group (2.3 ±â€Š0.5 days) than that in the control group (3.8 ±â€Š1.3 days, P = 0.03), but only 3 of 450 end of trial stool samples were identified as RV+ (<1%; CML, n = 2; control, n = 1). This hampered the assessment of efficacy of CML, despite the large a priori determined sample size. During the trial similar numbers of infants reported adverse events (AEs: CML 41%, control 46%), with the majority of events classified as mild and not related to the intervention. In conclusion, further clinical trials against a higher background of seasonal prevalence are necessary to assess efficacy of this nutritional intervention to prevent RVD. More important, however, high-ganglioside CML was acceptable for long-term consumption in infants ages 8 to 24 months.

Comparison of stool microbiota compositions, stool alpha1-antitrypsin and calprotectin concentrations, and diarrhoeal morbidity of Indonesian infants fed breast milk or probiotic/prebiotic-supplemented formula.

AIM: The composition of faecal microbiota of babies is known to be influenced by diet. Faecal calprotectin and α1-antitrypsin concentrations may be associated with mucosal permeability and inflammation. We aimed to assess whether there was any difference after consumption of a probiotic/prebiotic formula on faecal microbiota composition, calprotectin and α1-antitrypsin levels, and diarrhoea in comparison with breast milk-fed Indonesian infants. METHODS: One hundred sixty infants, 2 to 6 weeks old, were recruited to the study. They were either breastfed or formula fed (80 per group). Faecal samples were collected at recruitment and 3 months later. Bacterial groups characteristic of the human faecal microbiota were quantified in faeces by quantitative polymerase chain reaction. Calprotectin and α1-antitrypsin concentrations were measured using commercial kits. Details of diarrhoeal morbidity were documented and rated for severity. RESULTS: The compositions of the faecal microbiota of formula-fed compared with breast milk-fed children were similar except that the probiotic strain Bifidobacterium animalis subsp. lactis DR10 was more abundant after 3 months consumption of the formula. Alpha1-antitrypsin levels were higher in breastfed compared with formula-fed infants. The occurrence of diarrhoea did not differ between the groups of babies. CONCLUSION: Feeding Indonesian babies with a probiotic/prebiotic formula did not produce marked differences in the composition of the faecal microbiota in comparison with breast milk. Detrimental effects of formula feeding on biomarkers of mucosal health were not observed.

Bone health comparison in seven Asian countries using calcaneal ultrasound.

BACKGROUND: Bone density measurements by DXA are not feasible for large population studies, whereas portable ultrasound heel scanners can provide a practical way of assessing bone health status. The purpose of this study was to assess bone health in seven Asian countries using heel ultrasound. METHODS: Stiffness index (SI) was measured and T-scores generated against an Asian database were recorded for 598,757 women and 173,326 men aged over 21 years old using Lunar Achilles (GE Healthcare) heel scanners. The scanners were made available in public centres in Singapore, Vietnam, Malaysia, Taiwan, Thailand, Indonesia and the Philippines. RESULTS: The mean SI was higher for men than women. In women SI as well as T-scores declined slowly until approximately 45 years of age, then declined rapidly to reach a mean T-score of < -2.5 at about 71-75 years of age. For men, SI as well as the T-score showed a slow steady decline to reach a mean of -2.0 to -2.5 at about 81-85 years. The results for females indicate that there are differences in the rate of decline between countries (significant differences between the slopes at P < 0.05). Vietnam had the fastest decrease for both T-Score and SI, resulting in this population having the poorest bone health of all countries at older ages. The results for males aged 46-85 years indicate that there are no significant differences in the rate of decline between countries for SI and T-Score. In both men and women aged 46-85 years, Vietnam and Indonesia have the lowest SI as well as T-Score for all age groups. For Vietnam and Indonesia, more than 50% of the women could be at risk of having osteoporosis and related fractures after the age of 70, while in Thailand and the Philippines this was >80 years. CONCLUSIONS: The heel scan data shows a high degree of poor bone health in both men and women in Asian countries, raising concern about the possible increase in fractures with ageing and the expected burden on the public health system.

Effects of skim milk powder enriched with glycomacropeptide and G600 milk fat extract on frequency of gout flares: a proof-of-concept randomised controlled trial.

OBJECTIVES: Previous laboratory studies have identified two dairy fractions, glycomacropeptide (GMP) and G600 milk fat extract (G600), with anti-inflammatory effects in models of acute gout. The aim of this proof-of-concept clinical trial was to test the hypothesis that daily intake of skim milk powder (SMP) enriched with GMP and G600 can prevent gout flares. METHODS: This was a 3-month randomised double-blind controlled trial of milk products for prevention of gout flares. One hundred and twenty patients with recurrent gout flares were randomised to one of three arms: lactose powder control, SMP control and SMP enriched with GMP and G600 (SMP/GMP/G600). The primary end point was change in the frequency of gout flares using a daily flare diary measured monthly for 3 months. RESULTS: The frequency of gout flares reduced in all three groups over the 3-month study period compared with baseline. Over the 3-month study period there was a significantly greater reduction in gout flares in the SMP/GMP/G600 group (analysis of covariance p(group)=0.031, Tukey post hoc test compared with lactose control, p=0.044). Following treatment with SMP/GMP/G600 over the 3-month period, greater improvements were also observed in pain and fractional excretion of uric acid, with trends to greater improvement in tender joint count. Similar adverse event rates and discontinuation rates were observed between the three groups. CONCLUSIONS: This is the first reported controlled trial of dietary intervention in patients with gout, and suggests that SMP enriched with GMP and G600 may reduce the frequency of gout flares.

Fortified Milk Supplementation Improves Vitamin D Status, Grip Strength, and Maintains Bone Density in Chinese Premenopausal Women Living in Malaysia.

This study compared the effects of a high-calcium vitamin D fortified milk with added FOS-Inulin versus regular milk on serum parathyroid hormone (PTH), vitamin D status, grip strength (GS), as well as bone density in Chinese premenopausal women over 52 weeks. Premenopausal women (n = 133), mean age 41 (±5.1) years were randomized into control (n = 66; regular milk at 500 mg calcium per day) or intervention (Int; n = 67; fortified milk at 1200 mg calcium, 15 µg vitamin D, and 4 g FOS-Inulin per day) groups. Assessments were at baseline, weeks 12, 24, 36, and 52 for changes in vitamin D status, levels of PTH, and GS. Bone mineral densities (BMDs) of the lumbar spine (LS), femoral neck (FN), and whole body (WB) were assessed at baseline and week 52 using GE Lunar iDEXA (GE Healthcare, Madison, WI). At baseline, WB lean mass was positively associated with LS BMD (r = 0.30, p < 0.001) and FN BMD (r = 0.33, p = 0.003). Baseline 25(OH) vitamin D3 levels were 48.6 and 53.2 nmol/L (p = 0.57), respectively, and after the 12 months at 60.8 nmol/L (Int) versus 55.0 nmol/L (controls; p < 0.05 for change from baseline for both groups; no difference between groups at week 52). PTH levels decreased in both groups compared to baseline (p < 0.001), with no significant difference between groups. WB bone mineral content (BMC) and FN Z-score increased significantly in the Int group (p = 0.024 and p = 0.008). GS was positively associated with body weight, increasing in both groups over 52 weeks. Fortified milk improved vitamin D status, WB BMC, and Z-score of the FN, while regular milk maintained BMD. In addition, vitamin D status and GS improved.

Milk supplementation and bone health in young adult chinese women.

BACKGROUND: Milk is potentially beneficial for bone health, particularly for Chinese populations where consumption of dairy products is low. There are few data about milk consumption by Chinese women aged 20-35 years. This study examines whether milk supplement over 2 years caused increased bone mineral density (BMD) in Chinese women aged 20-35 years. METHODS: Four hundred forty-one community-living women living in Hong Kong SAR (221) and Beijing (220) China were randomized to receive milk supplement or nothing. The supplement consisted of two sachets of milk powder (1000 mg calcium, 80 microg vitamin K(1)), for 24 months. BMD at total hip, total spine, and whole body was measured at baseline and at 6, 12, 18, and 24 months; blood specimens were analyzed at baseline and at 3 and 24 months for biochemical indices of bone turnover and vitamin K. Urine samples also were collected. Analysis was by intention to treat as well as per protocol. Differences in change from baseline between the milk and control groups were analyzed using the mixed models approach to repeated measures, including the baseline value as a covariate. RESULTS: Both groups had an increase in BMD and a decrease in bone turnover markers over time, as an indicator of the process of attainment of peak bone mass during this period. Apart from a higher total spine BMD at 6 months in the milk group using per protocol analysis, there was little significant difference observed between the milk group and the control group. CONCLUSIONS: Age-related bone metabolism and lack of compliance most likely explain the lack of consistent changes in BMD or bone biochemical measures in response to milk supplementation for 2 years in Chinese women aged 20-35 years.

Effect of calcium fortified milk supplementation with or without vitamin K on biochemical markers of bone turnover in premenopausal women.

OBJECTIVE: We compared the effect of supplementation with a fortified skimmed milk product (high calcium skim milk) with or without added phylloquinone (vitamin K(1)) on markers of bone formation and resorption in premenopausal women. METHODS: Eighty-two women 20 to 35 y of age were randomly allocated to three groups. Two groups received two daily servings of high calcium skim milk (1000 mg/d of extra calcium) with or without added phylloquinone (80 microg/d) for 16 wk, and a third control group received no supplementation. Bone density was assessed at baseline and the bone markers, total osteocalcin, type I N-terminal procollagen peptide, and cross-linked C-telopeptide of type I collagen were measured at baseline and at weeks 2, 12, and 16. Serum phylloquinone and undercarboxylated osteocalcin were measured in the control and vitamin K-supplemented groups at weeks 0 and 16. RESULTS: Baseline values for age, body mass index, and bone density did not differ across groups. In vitamin K-supplemented women, mean serum phylloquinone concentrations increased from 0.27 to 0.76 microg/L (P < 0.05) and undercarboxylated osteocalcin concentrations decreased from 9.68 to 4.46 microg/L (P < 0.05) over 16 wk. Plasma cross-linked C-telopeptide of type I collagen, total osteocalcin, and type I N-terminal procollagen peptide levels decreased significantly in both supplemented groups compared with the control group over 16 wk (cross-linked C-telopeptide of type I collagen >30%, total osteocalcin and type I N-terminal procollagen peptide >15%). CONCLUSION: Fortified milk supplementation in premenopausal women reduced bone turnover significantly. Phylloquinone fortification substantially improved vitamin K status but had no demonstrable additive effect on bone turnover in this short-term study.

Encapsulated green kiwifruit extract: a randomised controlled trial investigating alleviation of constipation in otherwise healthy adults.

OBJECTIVES: Previous clinical trials have shown bowel function is improved through consumption of whole kiwifruit (Actinidia deliciosa). This study investigated whether encapsulated kiwifruit extract (1 g/day) could alleviate constipation in otherwise healthy adults. METHODS: Forty adults with confirmed constipation entered this trial, of which 32 completed with >80% compliance. Two capsules were self-administered morning and evening for 2 periods, each of 3 weeks duration, separated by a 3+ week washout in a double blind, randomised, placebo controlled crossover. Inclusion criteria included constipation with <=3 bowel movements (BM) per week. Daily records of defecation frequency and stool characteristics were obtained throughout treatment, as well as a measurement of gastrointestinal symptoms rating scale (GSRS) and quality of life (QoL) before and after each intervention arm. RESULTS: There was no difference in total BM over 3 weeks (p>0.05) or mean BM during each of weeks 1, 2 and 3 (p>0.05) between the kiwifruit extract and placebo when assessed from a faecal diary. There was also no detectable difference in defecation related scores of BM ease of defecation, volume, consistency or BM type assessed using Bristol stool chart scores. Nor was there a significant change in GSRS or QoL between pre and post treatment measures, when compared to placebo (p>0.05). CONCLUSIONS: This trial showed that improvement in bowel function or comfort was not achieved through supplementation with 1 g/day freeze dried kiwifruit extract. Efficacy from prior kiwifruit powder and whole fruit trials indicate that investigating higher doses of encapsulated kiwifruit extract may be worthwhile.

Postprandial metabolic responses of serum calcium, parathyroid hormone and C-telopeptide of type I collagen to three doses of calcium delivered in milk.

Acute doses of Ca rapidly increase serum Ca and reduce bone resorption concomitant with a reduction in serum parathyroid hormone (PTH) levels. The physiological response to a dose of Ca in milk and to a Ca salt may be different. The present study investigated Ca absorption patterns with increasing levels of fortification in milk, and the response to one dose of a Ca salt. A group of twenty-eight Asian women aged 20-45 years volunteered to attend the laboratory over several weeks. The fasted volunteers were randomised to one of three experimental drinks: 200 ml skimmed milk containing 250, 500 or 1000 mg Ca. A subgroup of seven volunteers also received a calcium gluconate/carbonate salt containing 1000 mg Ca in 200 ml water. Serial blood samples and urine were collected for 5 h from baseline. Different doses of Ca in milk resulted in a graded response in serum corrected Ca, PTH and C-telopeptide of type I collagen (CTx) but not ionised Ca. Serum Ca increased in response to all milk drinks and from 2 to 5 h the blood Ca levels were significantly different for the 250 and 1000 mg doses, as was the integrated response between the loads. The PTH response to the two higher doses was significantly more than following the 250 mg dose. The integrated response for CTx and urinary Ca between all three doses of Ca in milk was significantly different. A dose of Ca salt elicited a more immediate response reaching a plateau faster, and declining faster to baseline. Fortified milk is a safe matrix for delivering larger doses of Ca.

Dairy milk fat augments paclitaxel therapy to suppress tumour metastasis in mice, and protects against the side-effects of chemotherapy.

Milk fat is a natural product containing essential nutrients as well as fatty acids and other food factors with reported anti-cancer potential. Here bovine milk fat was tested for its ability to inhibit the growth of breast and colon cancers and their metastasis to the lung and liver; either alone or in combination with the chemotherapeutic agent paclitaxel. A diet containing 5% typical anhydrous milk fat (representing ~70% of the total dietary fat component) fed to Balb/c mice delayed the appearance of subcutaneous 4T1 breast and CT26 colon cancer tumours and inhibited their metastasis to the lung and liver, when compared to the control diet containing soybean oil as the only fat component. It augmented the inhibitory effects of paclitaxel on tumour growth and metastasis, and reduced the microvessel density of tumours. It displayed no apparent organ toxicity, but instead was beneficial for well-being of tumour-bearing mice by maintaining gastrocnemius muscle and epididymal adipose tissue that were otherwise depleted by cachexia. The milk fat diet ameliorated gut damage caused by paclitaxel in non-tumour-bearing mice, as evidenced by retention of jejunal morphology, villi length and intestinal γ-glutamyl transpeptidase activity, and inhibition of crypt apoptosis. It prevented loss of red and white blood cells due to both cancer-mediated immunosuppression and the cytotoxic effects of chemotherapy. The present study warrants the use of milk fat as an adjuvant to inhibit tumour metastasis during cancer chemotherapy, and to spare patients from the debilitating side-effects of cytotoxic drugs.

The effect of a fortified milk drink on vitamin D status and bone turnover in post-menopausal women from South East Asia.

Calcium and vitamin D are essential for bone growth; milk is an appropriate vehicle to be fortified with calcium, vitamin D and other minerals. The purpose of the current study was to compare the effect of supplementing with a high calcium milk drink with added vitamin D, magnesium and zinc (HCM) versus a placebo drink on serum parathyroid hormone (PTH) and vitamin D status as well as markers of bone formation/resorption in postmenopausal women living in South East Asia (Jakarta, Indonesia and Manila, the Philippines) over a period of 4 months. Calcium intake at baseline was 237 mg (median; 176-316, interquartile range) for Indonesia and 353 mg (median; 222-480, interquartile range) for the Filipino women per day. Fortified milk supplementation reduced the percentage of women that were insufficient in 25 (OH) vitamin D(3) (<50 nmol/L) from 70% to 22% in the Indonesian women and 20% to 0% in the Filipino women. Fortified milk supplementation significantly reduced parathyroid hormone levels (PTH) by week 2 (22% and 11%), C-telopeptide of type I collagen (CTX) by week 2 (34% and 27%), osteocalcin (OC) by week 8 (18% and 25%) and procollagen type I N-propeptide (PINP) by week 8 (15% and 21%), in women from Indonesia and the Philippines, respectively. Thus, the HCM intervention was able to significantly improve vitamin D status, lower PTH levels and reduce bone turnover in two groups of South East Asian women.

Heat-induced whey protein gels: protein-protein interactions and functional properties.

Heat-induced gelation (80 degrees C for 30 min or 85 degrees C for 60 min) of whey protein concentrate (WPC) solutions was studied using small deformation dynamic rheology, small and large deformation compression, and polyacrylamide gel electrophoresis (PAGE). The WPC solutions (15% w/w, pH 6.9) were prepared by dispersing WPC powder in water (control), 1% (w/w) sodium dodecyl sulfate (SDS) solution, and N-ethylmaleimide (NEM) solution at a protein/NEM molar ratio of 1:1 or in 10 mM dithiothreitol (DTT) solution. PAGE analyses showed that the heat treatment of control solutions contained both disulfide and non-covalent linkages between denatured protein molecules. Only disulfide linkages were formed in heated SDS-WPC solutions, whereas only non-covalent linkages were formed in DTT-WPC and NEM-WPC solutions during heating. In heated NEM-WPC solutions, the pre-existing disulfide linkages remained unaltered. Small deformation rheology measurements showed that the storage modulus (G') values, compared with those of the control WPC gels (approximately 14000 Pa), were 3 times less for the SDS-WPC gels (approximately 4000 Pa), double for the NEM-WPC gels (approximately 24000 Pa), and even higher for the DTT-WPC gels (approximately 30000 Pa). Compression tests suggested that the rubberiness (fracture strain) of the WPC gels increased as the degree of disulfide linkages within the gels increased, whereas the stiffness (modulus) of the gels increased as the degree of non-covalent associations among the denatured protein molecules increased.