Characterization and Mechanisms of Action of Avocado Extract Enriched in Mannoheptulose as a Candidate Calorie Restriction Mimetic.

Increased consumer interest in the avocado (Persea americana or Persea gratissima) has been attributed to established health benefits of this fruit associated with a wide range of ingredients. In search of effective calorie restriction mimetics (CRM), we present herein a consideration of possible health benefits of the rare sugar, mannoheptulose (MH), which acts as an intracellular glycolytic inhibitor and presents the highest concentration of this inhibitor in unripe avocados. A method for producing an extract of unripe avocado (AvX) to enrich concentrations of MH is described. Experiments using myocyte cultures demonstrated a pattern of CRM-like responses when treated with AvX. In vivo experiments confirmed that orally consumed AvX is bioavailable in both mice and dogs, as observed in urine and blood samples. Additional experiments in both these species demonstrated CRM-like improvements in glucose and insulin responses. In sum, the MH-enriched AvX exhibits promise as a CRM.

Understanding How Dogs Age: Longitudinal Analysis of Markers of Inflammation, Immune Function, and Oxidative Stress.

As in human populations, advances in nutrition and veterinary care have led to an increase in the lifespan of companion animals. Detrimental physiological changes occurring later in life must be understood before interventions can be made to slow or reduce them. One important aspect of human aging is upregulation of the inflammatory response and increase in oxidative damage resulting in pathologies linked to chronic inflammation. To determine whether similar processes occur in the aging dog, changes in markers of inflammation and oxidative stress were investigated in 80 Labrador retrievers from adulthood to the end of life. Serum levels of immunoglobulin M (p < .001) and 8-hydroxy-2-deoxyguanosine (p < .001) increased with age, whereas no effect of age was detected for immunoglobulin G or C-reactive protein unless the last year of life was included in the analysis (p = .002). Baseline levels of heat shock protein 70 decreased with age (p < .001) while those after exposure to heat stress were maintained (p = .018). However, when excluding final year of life data, a decline in the heat shock protein 70 response after heat stress was observed (p = .004). These findings indicate that aging dogs undergo changes similar to human inflammaging and offer the possibility of nutritional or pharmacological intervention to delay or reduce these effects.

Astaxanthin stimulates cell-mediated and humoral immune responses in cats.

Astaxanthin is a potent antioxidant carotenoid and may play a role in modulating immune response in cats. Blood was taken from female domestic shorthair cats (8-9 mo old; 3.2 ± 0.04 kg body weight) fed 0, 1, 5 or 10mg astaxanthin daily for 12 wk to assess peripheral blood mononuclear cell (PBMC) proliferation response, leukocyte subpopulations, natural killer (NK) cell cytotoxic activity, and plasma IgG and IgM concentration. Cutaneous delayed-type hypersensitivity (DTH) response against concanavalin A and an attenuated polyvalent vaccine was assessed on wk 8 (prior to vaccination) and 12 (post-vaccination). There was a dose-related increase in plasma astaxanthin concentrations, with maximum concentrations observed on wk 12. Dietary astaxanthin enhanced DTH response to both the specific (vaccine) and nonspecific (concanavalin A) antigens. In addition, cats fed astaxanthin had heightened PBMC proliferation and NK cell cytotoxic activity. The population of CD3(+) total T and CD4(+) T helper cells were also higher in astaxanthin-fed cats; however, no treatment difference was found with the CD8(+) T cytotoxic and MHC II(+) activated lymphocyte cell populations. Dietary astaxanthin increased concentrations of plasma IgG and IgM. Therefore, dietary astaxanthin heightened cell-mediated and humoral immune responses in cats.

Dietary fish oil and flaxseed oil suppress inflammation and immunity in cats.

The modulatory activity of dietary n-3 fatty acids on inflammation and immune response in domestic cats is unknown. Mature female cats (n=14/treatment) were fed control, fish oil or flaxseed oil diets with n-6:n-3 fatty acid ratios of 20:1, 5:1 and 5:1, respectively, for 12 wk. Immune response was assessed on wk 0, 6 and 12, and skin hypersensitivity response on wk 6 and 12. Fish oil increased (P<0.01) eicosapentaenoic and docosahexaenoic acids in plasma and skin, whereas flaxseed oil increased α-linolenic acid. Fish and flaxseed oils decreased (P<0.01) skin inflammatory response to histamine. Cats fed fish but not flaxseed oil had higher (P<0.05) skin leukotriene LTB(5), but not LTB(4). Fish and flaxseed oils lowered B, total T and T(h) subset populations, and leukocyte proliferative response to PWM (P<0.05). In contrast, there was no change in ConA- or PHA-induced lymphocyte proliferation, Tc and MHC II cell populations, DTH response, NK cytotoxicity, IL-2 production, or plasma IgG concentrations. Therefore, fish and flaxseed oil can reduce skin inflammatory responses in cats, however, flaxseed oil appears less immunosuppressive than fish oil.

Dietary astaxanthin enhances immune response in dogs.

No information is available on the possible role of astaxanthin on immune response in domestic canine. Female Beagle dogs (9-10 mo old; 8.2 ± 0.2 kg body weight) were fed 0, 10, 20 or 40 mg astaxanthin daily and blood sampled on wk 0, 6, 12, and 16 for assessing the following: lymphoproliferation, leukocyte subpopulations, natural killer (NK) cell cytotoxicity, and concentrations of blood astaxanthin, IgG, IgM and acute phase proteins. Delayed-type hypersensitivity (DTH) response was assessed on wk 0, 12 and 16. Plasma astaxanthin increased dose-dependently and reached maximum concentrations on wk 6. Dietary astaxanthin enhanced DTH response to vaccine, concanavalin A-induced lymphocyte proliferation (with the 20mg dose at wk 12) and NK cell cytotoxic activity. In addition, dietary astaxanthin increased concentrations of IgG and IgM, and B cell population. Plasma concentrations of C reactive protein were lower in astaxanthin-fed dogs. Therefore, dietary astaxanthin heightened cell-mediated and humoral immune response and reduced DNA damage and inflammation in dogs.

Astaxanthin uptake in domestic dogs and cats.

BACKGROUND: Research on the uptake and transport of astaxanthin is lacking in most species. We studied the uptake of astaxanthin by plasma, lipoproteins and leukocytes in domestic dogs and cats. METHODS: Mature female Beagle dogs (18 to 19 mo old; 11 to 14 kg BW) were dosed orally with 0, 0.1, 0.5, 2.5, 10 or 40 mg astaxanthin and blood taken at 0, 3, 6, 9, 12, 18 and 24 h post-administration (n = 8/treatment). Similarly, mature domestic short hair cats (12 mo old; 3 to 3.5 kg body weight) were fed a single dose of 0, 0.02, 0.08, 0.4, 2, 5, or 10 mg astaxanthin and blood taken (n = 8/treatment) at the same interval. RESULTS: Both dogs and cats showed similar biokinetic profiles. Maximal astaxanthin concentration in plasma was approximately 0.14 mumol/L in both species, and was observed at 6 h post-dosing. The plasma astaxanthin elimination half-life was 9 to 18 h. Astaxanthin was still detectable by 24 h in both species. In a subsequent study, dogs and cats were fed similar doses of astaxanthin daily for 15 to 16 d and astaxanthin uptake by plasma, lipoproteins, and leukocytes studied. In both species, plasma astaxanthin concentrations generally continued to increase through d 15 or 16 of supplementation. The astaxanthin was mainly associated with high density lipoprotein (HDL). In blood leukocytes, approximately half of the total astaxanthin was found in the mitochondria, with significant amounts also associated with the microsomes and nuclei. CONCLUSION: Dogs and cats absorb astaxanthin from the diet. In the blood, the astaxanthin is mainly associated with HDL, and is taken up by blood leukocytes, where it is distributed to all subcellular organelles. Certain aspects of the biokinetic uptake of astaxanthin in dogs and cats are similar to that in humans.

Exploring mechanisms of sex differences in longevity: lifetime ovary exposure and exceptional longevity in dogs.

To move closer to understanding the mechanistic underpinnings of sex differences in human longevity, we studied pet dogs to determine whether lifetime duration of ovary exposure was associated with exceptional longevity. This hypothesis was tested by collecting and analyzing lifetime medical histories, age at death, and cause of death for a cohort of canine 'centenarians'--exceptionally long-lived Rottweiler dogs that lived more than 30% longer than average life expectancy for the breed. Sex and lifetime ovary exposure in the oldest-old Rottweilers (age at death, > or = 13 years) were compared to a cohort of Rottweilers that had usual longevity (age at death, 8.0-10.8 years). Like women, female dogs were more likely than males to achieve exceptional longevity (OR, 95% CI = 2.0, 1.2-3.3; P = 0.006). However, removal of ovaries during the first 4 years of life erased the female survival advantage. In females, a strong positive association between ovaries and longevity persisted in multivariate analysis that considered other factors, such as height, body weight, and mother with exceptional longevity. A beneficial effect of ovaries on longevity in females could not be attributed to resistance against a particular disease or major cause of death. Our results document in dogs a female sex advantage for achieving exceptional longevity and show that lifetime ovary exposure, a factor not previously evaluated in women, is associated with exceptional longevity. This work introduces a conceptual framework for designing additional studies in pet dogs to define the ovary-sensitive biological processes that promote healthy human longevity.

Isolation and characterization of primary canine lens epithelial cells.

OBJECTIVE: The purpose of this study was to characterize the proliferation and differentiation of primary canine lens epithelial cells (LEC) under standard culture conditions. PROCEDURE: Canine LEC were isolated by mechanical dissection of the canine globe and enzymatic digestion of the lens capsule from fresh lenses. Isolated capsules and cell suspensions were seeded in laminin-coated culture flasks. Canine LEC proliferated and formed monolayers, which could be passaged and maintained for approximately 2 weeks. Cells were characterized morphologically and cell lysates examined for expression of protein markers of epithelial origin and differentiation. RESULTS: Canine LEC exhibit morphologic characteristics of epithelial cells when cultured on laminin/lysine coated flasks. Expression of epithelial cell marker, cytokeratin 5, was highest at passage 1 and diminished with increasing passage number. Expression of gamma-crystallin, a protein found only in differentiated lens fiber cells, increased at passage 6. A laminin/lysine-coated surface supported optimal proliferation of canine LEC. Both an initial seeding density of 1 x 10(5) cells/cm(2) and culture in Dulbecco's modified essential media (DMEM) supplemented with 10% FBS supported a doubling time of less than 48 h in canine LEC. CONCLUSION: This study demonstrates that primary canine LEC retain the characteristics of lens epithelial cells prior to passage 6 under the described culture conditions and represent a suitable in vitro model for investigating lens physiology and cataractogenesis.

Modulation of immune response through nutraceutical interventions: implications for canine and feline health.

Mounting research demonstrates that certain nutraceutical compounds interact with the immune system. These interactions may be positive or negative depending on the compound or dose administered to the individual. Understanding the mechanisms by which these compounds work should provide opportunities to design nutritional interventions to bolster the health of dogs and cats.

Effect of age on immune parameters and the immune response of dogs to vaccines: a cross-sectional study.

The evaluation of anti-aging intervention strategies in dogs would benefit from reliable quantitative biomarkers of aging. In the present study, the expression of various immune parameters was measured in young and old dogs to identify potential biomarkers of aging. The second goal of the study was to determine the effect of age on the immune response to vaccines. The immune function, including the antibody response to vaccines, was determined in 32 young adult (3.15+/-0.8 years of age) and 33 old dogs (12.1+/-1.3 years of age) of various breeds. Old dogs had a significantly lower lymphocyte proliferative response and a lower percentage of CD4+ T cells and CD45R+/CD4+ T cells, and a higher percentage of CD8+ T cells and a higher concentration of serum and salivary IgA. The most significant differences (P<0.001) occurred in the lymphocyte proliferative responses to ConA and PHA, the CD4:CD8 ratio, and the percentage of CD45R+/CD4+ T cells suggesting that these parameters are potential biomarkers of aging. There was no difference in the percentage of total T and B lymphocytes and the concentration of serum IgM and IgG. Both groups of dogs had protective titers against distemper virus, parvovirus and rabies virus before annual revaccination. The pre-vaccination titer against rabies virus was higher in the old dogs than in the young dogs, and there were no differences in post-vaccination titers against any of the viruses. This suggests that annual vaccination protocols provide adequate protection for old dogs.

Exceptional longevity in pet dogs is accompanied by cancer resistance and delayed onset of major diseases.

To characterize extreme aged pet dogs as a first step in developing an animal model of exceptional longevity, we constructed lifetime medical histories for 345 Rottweiler dogs using information collected from owners and veterinarians. Extreme aged dogs (alive at the 95th percentile age at death for the study population, > or =13.3 years) were compared with a usual longevity group (9-10 years). Exceptional longevity in Rottweiler dogs was accompanied by a significant delay in the onset of major life-threatening diseases; 76% of extreme aged dogs remained free of all major diseases during the first 9 years of life. Only 19% of extreme aged dogs died of cancer versus 82% of dogs with usual longevity (p <.0001). The reduction in cancer mortality in oldest-old pet dogs mimics that seen in human centenarians and provides strong rationale for using this animal model to study comparative mechanisms of cancer resistance in the extreme aged.

Effects of age and dietary beta-carotene on immunological variables in dogs.

beta-Carotene is a naturally occurring carotenoid reported to have health-promoting effects in several species. Advancing age is known to have a negative impact on various immune variables in several species. This study was conducted in order to assess the effect of age on immune response in dogs and to determine whether beta-carotene is able to reverse this age-associated decline. To test this hypothesis, young and old dogs (n = 36) were fed either a control diet or experimental diets containing supplemental beta-carotene for 2-month periods. Age significantly (P < .05) lowered CD4+ T cell populations (47.2% versus 33.7%; young-control versus old-control, respectively) and beta-carotene restored percent distributions in old dogs to nonsignificance versus younger controls (41.0%). T cell proliferation was lower in old dogs (30,254 +/- 2,248 versus 14,811 +/- 2,497 cCPM; young-control versus old-control, respectively; P < .05), and beta-carotene supplementation significantly improved responses in this age group (21,329 +/- 2,275 cCPM). Although B cell proliferation was depressed with age (17,967 +/- 1,384 versus 7,535 +/- 1,469 cCPM; young-control versus old-control, respectively; P < .05), beta-carotene supplementation improved B cell proliferation in young dogs (23,500 +/- 1,339 cCPM). Old dogs displayed lower delayed-type hypersensitivity test (DTH) responses versus younger controls to both phytohemagglutinin-P (PHA; 11.1 +/- 0.95 versus 7.57 +/- 1.15 mm; young-control versus old-control, respectively; P < .05) and sheep red blood cell (RBC; 9.12 +/- 0.62 versus 8.08 +/- 0.75 mm; young-control versus old-control, respectively; P < .10). beta-Carotene improved these responses, mostly within the first 24-48 hours after injection. In summary, older dogs have lower immunological responses compared with younger controls. beta-Carotene supplementation significantly restored immune responses in older dogs when compared with their age-matched controls and younger counterparts.

Oxidized lipid depresses canine growth, immune function, and bone formation.

Dietary oxidized lipids can increase oxidative stress and potentially contribute to a variety of disease syndromes. This research describes the first use of a canine model to assess the effects of dietary oxidized lipids on growth, antioxidant status, and some immune functions. Three groups of eight, two-month old coon-hound puppies were pair fed diets for 16 weeks. The control diet contained <50 ppm aldehydes, and two additional diets contained thermally oxidized lipids targeted to contain 100 ppm aldehydes (medium-oxidation) and 500 ppm aldehydes (high-oxidation). Dogs fed the high-oxidation diet weighed less than those from the medium-oxidation (P < 0.05) and control groups (P < 0.001) at the end of the study. Oxidized lipids reduced serum vitamin E levels, total body fat content, and bone appositional rate. At different time points of the study, peripheral blood neutrophils and monocytes from dogs fed the HO diet had reduced oxidative burst capacity and produced less superoxide and hydrogen peroxide when stimulated with phorbol esters compared to the control group. Lymphocyte blastogenesis in response to concanavalin A was suppressed by dietary oxidized lipid. This study indicates that dietary oxidized lipids negatively affect the growth, antioxidant status, and some immune functions of dogs. Importantly, some effects are evident at 100 ppm aldehydes in the diet, which is a moderate level of oxidation. The rapid growth and weight gain of the dog during the first 6 months of life may also provide a better model for assessing the risks of dietary oxidized lipid in children and adolescents than previously used rodent models.