Kate Palmano

‘Iron-saturated’ lactoferrin is a potent natural adjuvant for augmenting cancer chemotherapy

Bovine lactoferrin (bLf), an iron‐containing natural defence protein found in bodily secretions, has been reported to inhibit carcinogenesis and the growth of tumours. Here, we investigated whether natural bLf and iron‐saturated forms of bLf differ in their ability to augment cancer chemotherapy. bLf was supplemented into the diet of C57BL/6 mice that were subsequently challenged subcutaneously with tumour cells, and treated by chemotherapy. Chemotherapy eradicated large (0.6 cm diameter) EL‐4 lymphomas in mice that had been fed iron‐saturated bLf (here designated Lf+) for 6 weeks prior to chemotherapy, but surprisingly not in mice that were fed lesser iron‐saturated forms of bLf, including apo‐bLf (4% iron saturated), natural bLf (∼15% iron saturated) and 50% iron‐saturated bLf. Lf+‐fed mice bearing either EL‐4, Lewis lung carcinoma or B16 melanoma tumours completely rejected their tumours within 3 weeks following a single injection of either paclitaxel, doxorubicin, epirubicin or flurouracil, whereas mice fed the control diet were resistant to chemotherapy. Lf+ had to be fed to mice for more than 2 weeks prior to chemotherapy to be wholly effective in eradicating tumours from all mice, suggesting that it acts as a competence factor. It significantly reduced tumour vascularity and blood flow, and increased antitumour cytotoxicity, tumour apoptosis and the infiltration of tumours by leukocytes. Lf+ bound to the intestinal epithelium and was preferentially taken up within Peyers patches. It increased the production of Th1 and Th2 cytokines within the intestine and tumour, including TNF, IFN‐γ, as well as nitric oxide that have been reported to sensitize tumours to chemotherapy. Importantly, it restored both red and white peripheral blood cell numbers depleted by chemotherapy, potentially fortifying the mice against cancer. In summary, bLf is a potent natural adjuvant and fortifying agent for augmenting cancer chemotherapy, but needs to be saturated with iron to be effective.

Acute effect of milk on serum urate concentrations: a randomised controlled crossover trial

Objectives Recent observational studies have highlighted the beneficial role of dairy ingestion in gout prevention. The aims of this study were to determine the acute effects of milk ingestion on serum urate concentrations and examine the mechanisms of these effects. Methods This was a short-term randomised controlled crossover trial of milk in 16 healthy male volunteers. The following products were tested (each 80 g protein): soy control, early season skim milk, late season skim milk (containing high concentrations of orotic acid, a naturally occurring uricosuric agent) and ultrafiltrated MPC 85 skim milk. Each participant received a single dose of each product in random order. Serum and urine were obtained immediately before and then hourly over a 3 h period after ingestion of each study product. Results Ingestion of the soy control led to an increase in serum urate concentrations by approximately 10%. In contrast, ingestion of all milks led to a decrease in serum urate concentrations by approximately 10% (p<0.0001). All products (including soy) rapidly increased the fractional excretion of uric acid (FEUA). Late season milk led to a greater increase in FEUA than MPC 85 (p=0.02) and early season milk (p=0.052). There were no differences over time in serum oxypurines or purine-containing nucleosides. However, all products increased the fractional excretion of xanthine. Conclusions Intact milk has an acute urate-lowering effect. These data provide further rationale for long-term intervention studies to determine whether such dietary interventions have an adjunctive role in the management of individuals with hyperuricaemia and gout.

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 pgroup=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.

The Role of Gangliosides in Neurodevelopment

Gangliosides are important components of neuronal cell membranes and it is widely accepted that they play a critical role in neuronal and brain development. They are functionally involved in neurotransmission and are thought to support the formation and stabilization of functional synapses and neural circuits required as the structural basis of memory and learning. Available evidence, as reviewed herein, suggests that dietary gangliosides may impact positively on cognitive functions, particularly in the early postnatal period when the brain is still growing. Further, new evidence suggests that the mechanism of action may be through an effect on the neuroplasticity of the brain, mediated through enhanced synaptic plasticity in the hippocampus and nigro-striatal dopaminergic pathway.

The effect of whey acidic protein fractions on bone loss in the ovariectomised rat

Bovine milk has been shown to contain bioactive components with bone-protective properties. Earlier studies on bovine milk whey protein showed that it suppressed bone resorption in the female ovariectomised rat. A new osteotropic component was subsequently identified in the whey basic protein fraction, but bone bioactivity may also be associated with other whey fractions. In the present study, we investigated whether acidic protein fractions isolated from bovine milk whey could prevent bone loss in mature ovariectomised female rats. Six-month-old female rats were ovariectomised (OVX) or left intact (sham). The OVX rats were randomised into four groups. One group remained the control (OVX), whereas three groups were fed various whey acidic protein fractions from milk whey as 3 g/kg diet for 4 months. Outcomes were bone mineral density, bone biomechanics and markers of bone turnover. Bone mineral density of the femurs indicated that one of the whey AF over time caused a recovery of bone lost from OVX. Plasma C-telopeptide of type I collagen decreased significantly in all groups except OVX control over time, indicating an anti-resorptive effect of whey acidic protein. Biomechanical data showed that the AF may affect bone architecture as elasticity was increased by one of the whey AF. The femurs of AF-supplemented rats all showed an increase in organic matter. This is the first report of an acidic whey protein fraction isolated from milk whey that may support the recovery of bone loss in vivo.

Identification of dairy fractions with anti-inflammatory properties in models of acute gout

Aims Large epidemiological studies have shown that low-fat dairy intake reduces the risk of developing gout. It was hypothesised that factors within dairy fractions inhibit the inflammatory response to monosodium urate monohydrate (MSU) crystals. Methods Dairy fractions were tested in MSU crystal-stimulated THP-1 cell assays. Fractions with inhibitory effects were then tested in the murine urate peritonitis model. Results Two dairy fractions were found to have consistent inhibitory effects. Glycomacropeptide (GMP) and G600 milk fat extract both inhibited interleukin-1β (IL1β) gene and protein expression in the THP-1 cell assay. Conversely, standard milk fat increased IL8 protein expression in the THP-1 cell assay. Oral administration of GMP and G600 milk fat extract inhibited cellular influx in the urate peritonitis model. Conclusions Both protein and lipid fractions within dairy products are capable of modulating the inflammatory response to MSU crystals.

Effects of Dairy Intake on Hyperuricemia and Gout

Dietary modification is frequently recommended for patients with gout. Longitudinal observational studies have shown a clear inverse relationship between low-fat dairy intake and gout risk. Several checkpoints in gout pathogenesis may be targeted by dairy intake. Cross-sectional and short-term intervention studies of healthy volunteers have demonstrated that low-fat dairy intake has a moderate urate-lowering effect. In addition, certain dairy fractions, particularly glycomacropeptide and G600 milk fat extract, have anti-inflammatory properties in experimental models of acute gout. Such anti-inflammatory properties may contribute to the reduction in gout risk through inhibition of the inflammatory response to monosodium urate crystals within the joint. Well-controlled intervention studies in patients with gout are now needed to determine the clinical relevance of these observations in order to guide dietary recommendations for this disease.

“Iron-saturated” bovine lactoferrin improves the chemotherapeutic effects of tamoxifen in the treatment of basal-like breast cancer in mice

BackgroundTamoxifen is used in hormone therapy for estrogen-receptor (ER)-positive breast cancer, but also has chemopreventative effects against ER-negative breast cancers. This study sought to investigate whether oral iron-saturated bovine lactoferrin (Fe-Lf), a natural product which enhances chemotherapy, could improve the chemotherapeutic effects of tamoxifen in the treatment of ER-negative breast cancers.MethodsIn a model of breast cancer prevention, female Balb/c mice treated with tamoxifen (5 mg/Kg) were fed an Fe-Lf supplemented diet (5 g/Kg diet) or the base diet. At week 2, 4T1 mammary carcinoma cells were injected into an inguinal mammary fat pad. In a model of breast cancer treatment, tamoxifen treatment was not started until two weeks following tumor cell injection. Tumor growth, metastasis, body weight, and levels of interleukin 18 (IL-18) and interferon γ (IFN-γ) were analyzed.ResultsTamoxifen weakly (IC50 ~ 8 μM) inhibited the proliferation of 4T1 cells at pharmacological concentrations in vitro. In the tumor prevention study, a Fe-Lf diet in combination with tamoxifen caused a 4 day delay in tumor formation, and significantly inhibited tumor growth and metastasis to the liver and lung by 48, 58, and 66% (all P < 0.001), respectively, compared to untreated controls. The combination therapy was significantly (all P < 0.05) more effective than the respective monotherapies. Oral Fe-Lf attenuated the loss of body weight caused by tamoxifen and cancer cachexia. It prevented tamoxifen-induced reductions in serum levels of IL-18 and IFN-γ, and intestinal cells expressing IL-18 and IFN-γ. It increased the levels of Lf in leukocytes residing in gut-associated lymphoid tissues. B, T and Natural killer (NK) cells containing high levels of Lf were identified in 4T1 tumors, suggesting they had migrated from the intestine. Similar effects of Fe-Lf and tamoxifen on tumor cell viability were seen in the treatment of established tumors.ConclusionsThe results indicate that Fe-Lf is a potent natural adjuvant capable of augmenting the chemotherapeutic activity of tamoxifen. It could have application in delaying relapse in tamoxifen-treated breast cancer patients who are at risk of developing ER-negative tumors.

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 under 5-yrs, presenting commonly with diarrhoeal symptoms. In a prospective 12-wk 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, aged 8-24 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, n = 20) and throughout the trial, with only 110 total episodes of ACD over 12-wks (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 (2.3 ± 0.5 days) group than Control (3.8 ± 1.3 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 aginst a higher background of seasonal prevalence are necessary to assess efficacy of this nutritional intervention to prevent RVD. Importantly, however, high-ganglioside CML was acceptable for long-term consumption in infants aged 8-24 months.ABSTRACTRotavirus (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 ± 0.5 days) than that in the control group (3.8 ± 1.3 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.

Lactoferrin – A Potential Anabolic Intervention in Osteoporosis

Osteoporosis or porous bone was first described by Fuller Albright approximately 70 years ago as having “too little bone in the bone”. Bone tissue is maintained throughout life by being continually replaced and in osteoporosis bone resorption exceeds bone formation resulting in bone loss. The majority of current treatments for osteoporosis are antiresorptive, decreasing osteoclast activity and preventing further bone loss. Therapeutic agents that activate osteoblasts and increase bone formation have the potential benefit of restoring bone rather than only preventing further deterioration, but only a small number of safe anabolic therapies are currently available. Milk is a rich biological fluid that contains many growth factors and provides nutrition at a time of very rapid skeletal growth and development in the neonate, and was therefore considered as a possible source of factors with anabolic effects on bone. Investigations of fractions of whey proteins extracted from milk identified lactoferrin as a bone-active factor. Lactoferrin is an iron-binding glycoprotein which as well as being present in milk is found in other epithelial secretions. It is a pleiotropic factor with potent antimicrobial and immunomodulatory activities, and shows anabolic effects in bone at physiological concentrations. In a number of recent studies in humans and experimental animals dietary lactoferrin supplementation improved bone mineral density, bone markers and bone strength. The current chapter discusses the structure and function of lactoferrin, the bone-effects of lactoferrin in vitro and in vivo, and the potential use of lactoferrin for the improvement of bone health.