Effect of maternal overnutrition on predisposition to insulin resistance in the foal: Foal skeletal muscle development and insulin signaling.

Skeletal muscle plays an integral role in the ability of a horse to perform at high levels. Shifts in skeletal muscle development in response to maternal plane of nutrition may have substantial and lasting impacts on athletic performance and whole-body metabolism. Therefore, sixteen Quarter Horse mares were used in a completely randomized design and maintained at a body condition score (BCS) 6 until start of third trimester. On d 235 of gestation, mares were randomly assigned to receive one of two dietary treatments with a diet formulated to meet requirements during late gestation (CON; n = 8), and an overfed diet (HIGH; n = 8) where mares received an additional 40% above CON. Five h after parturition, foals were euthanized, and gluteus medius, triceps brachii, and semitendinosus were harvested for analyses. Gene expression was determined by qPCR and western immunoblotting was used to quantify total and phosphorylated forms of proteins involved in skeletal muscle metabolism with tubulin as the loading control. All data were analyzed using PROC MIXED of SAS. Foals from HIGH mares exhibited larger skeletal muscle fibers by area (P <0.05), and a shift in muscle fiber development towards type I slow twitch muscle fibers (P <0.05). Relative expression of glucose transporter 4 (GLUT4) was lower in HIGH foals compared to CON in gluteus medius (P = 0.05). Insulin receptor isoform B (INSR-B) and insulin-like growth factor 1 receptor (IGF1R) were greater in triceps brachii of HIGH foals compared to CON (P ≤ 0.03). Insulin receptor isoform A (INSR-A), however, tended to be lower in triceps brachii of HIGH compared to CON (P = 0.10). Ratios of phosphorylated to total extracellular signal-regulated protein kinase 1/2 (ERK1/2) and c-June N-terminal kinase (JNK) were higher in HIGH foals compared to CON (P ≤0.04) in gluteus medius. There were no differences observed for phosphorylated to total protein ratios in semitendinosus and triceps brachii muscles; however, total ERK1/2 tended to be elevated (P <0.10) in semitendinosus from CON foals compared to HIGH. There was no difference in phosphorylated or total protein kinase B (AKT) (P >0.14). These data indicate hypertrophy of skeletal muscle fibers and a shift towards type I slow twitch fibers in HIGH foals. Furthermore, this study identifies muscle specific changes in gene expression and downstream insulin receptor signaling, which may contribute to future metabolic abnormalities in response to maternal overnutrition.

Effect of bioactive proteins on gait kinematics and systemic inflammatory markers in mature horses.

Twenty-seven mature Quarter horses were used in a randomized design to determine the effects of bioactive protein supplementation on gait kinematics and systemic inflammatory markers in a 34-d trial. Treatments consisted of oral doses of 230 g/d of pelleted supplements containing 0 g (CON; n = 9), 40 g of bioactive protein (40BP; n = 9; LIFELINE, APC, LLC, Ankeny, IA), and 80 g of bioactive protein (80BP; n = 9) daily. Horses were fed a commercial concentrate at 0.5% BW (as-fed) and received ad libitum coastal bermudagrass (Cynodon dactylon) hay daily. On day 33, horses consistent in exercise (CON, n = 6; 40BP, n = 8; 80BP, n = 7) participated in a trailering and riding challenge. Kinematic gait analysis was performed on day 0 for use as a covariate, and on day 14, 28, and 34 to allow for the determination of potential time and dosage effects. Video footage was collected and analyzed using gait analysis software (EquineTec, Monroe, GA) for the determination of stride length (SL) and range of motion (ROM). Blood was collected via jugular venipuncture on days 0, 14, 28, and 34 for determination of systemic expression of tumor necrosis factor (TNF)-α and IL-1ß. Data were analyzed using PROC MIXED of SAS. A trend towards treatment × time interaction was observed in ROM of the knee at the walk (P = 0.10), due to the increasing ROM for 40BP and 80BP as time increased and decreasing ROM for CON. A treatment × time interaction was observed (P < 0.01) for hock ROM at a walk resulting from CON and 80BP decreasing from day 14 to 28 with 40BP increasing, while from day 28 to 34 ROM at a walk decreased for 40BP and increased for 80BP. The main effect of treatment on hock ROM at the walk was quadratic (P < 0.01) and characterized by higher ROM values for 40BP compared to CON or 80BP. Dietary treatment lengthened (P = 0.04) SL of the hind limb at the walk for 40BP and 80BP compared to CON on both days 14 and 28. A significant treatment × time interaction was observed in the expression of IL-1ß (P < 0.01) and can be explained by lower concentrations of IL-1ß for 80BP on day 34 compared to the other treatments, with 40BP being intermediate and CON being the highest. Increased articular ROM with decreased expression of IL-1ß may indicate potential anti-inflammatory effects of 80 g/d of bioactive proteins.

Effect of maternal overnutrition on predisposition to insulin resistance in the foal: Maternal parameters and foal pancreas histoarchitecture.

Results from previous studies indicate that maternal overnutrition during late gestation predisposes foals to metabolic disease, however, specific mechanisms resulting in disease remain unknown. Quarter Horse mares (n = 16), were randomly assigned to dietary treatments, beginning on gestational day 235, and consisted of a control group (CON- diet meeting nutrient requirement; n = 8) or an overfed diet (HIGH; n = 8) where mares received an additional 40 % above CON. On gestational days 285 and 315, an intravenous glucose tolerance test (FSIGTT) was conducted. Following parturition, foals were separated from the mare, prohibited from nursing, and an FSIGTT was conducted at 2 h postpartum. Foals were immediately euthanized and tissues preserved for analyses. There was no effect of treatment on foal BW (P = 0.50), pancreas weight (P = 0.60), or FSIGTT area under the curve for glucose (P = 0.80) and insulin (P = 0.70). Colocalization of α-amylase to isolate pancreatic islets of Langerhans indicated increased islet number and size in foals from HIGH mares (P < 0.01). Immunofluoresent analysis of insulin, glucagon, and somatostatin indicate no difference in intensity of staining (P> 0.10). Foals exposed to overnutrition during peak fetal growth had altered pancreatic islet development that may lead to adult-onset metabolic disease.

Age-related effects on markers of inflammation and cartilage metabolism in response to an intra-articular lipopolysaccharide challenge in horses.

Eighteen Quarter Horses were used in a randomized complete design for a 28-d experiment to evaluate age-related effects on inflammation and cartilage turnover after induction of a single inflammatory insult using lipopolysaccharide (LPS). Horses were grouped by age as yearlings (3 males and 3 females), 2 to 3 yr olds (2/3 yr old; 2 males and 4 females), and skeletally mature 5 to 8 yr olds (mature; 2 males and 4 females). On d 0, all horses were individually housed and fed diets that met or exceeded requirements. On d 14, horses were challenged with an intra-articular injection of LPS. Radial carpal joints were randomly assigned to receive 0.5 ng LPS solution obtained from O55:B5 or 0.8 mL sterile lactated Ringer's solution as a contralateral control. Synovial fluid was collected prior to LPS injection at h 0 before injection and at 6, 12, 24, 168, and 336 h after injection. Samples were analyzed using commercial ELISA kits for PGE, collagenase cleavage neoepitope (C2C), and carboxypropeptide of type II collagen (CPII). Heart rate (HR), respiratory rate (RR), and rectal temperature (RT) were monitored over the initial 24 h and carpal circumference and surface temperature were also recorded, with additional measurements at 168 and 336 h. Data were analyzed using PROC MIXED of SAS. Values for RT, HR, and RR were within the normal range for each age group. Heart rate and RT were influenced by age ( < 0.01), whereas RR was unaffected ( ≤ 0.21). Joint circumference was not influenced by age of horse ( = 0.84), but circumference and surface temperature increased ( < 0.01) over time across all age groups. Synovial PGE concentrations tended ( = 0.09) to be influenced by age, with yearlings having lower ( = 0.03) concentrations than mature horses. Concentrations of synovial C2C were affected by age of horse, with yearlings and 2/3 yr olds having lower ( < 0.01) concentrations than mature horses. Similarly, synovial CPII was influenced by age, with yearlings and 2/3 yr olds having lower ( ≤ 0.02) concentrations than mature horses. Ratios of anabolic CPII to catabolic C2C varied by age, with mature and 2/3-yr-old horses having greater ( < 0.01) values compared with yearlings. These results indicate that inflammation and the corresponding cartilage turnover in response to LPS administration vary with age.

Influence of short-term dietary starch inclusion on the equine cecal microbiome.

The objective of this study was to determine bacterial community profiles of the equine cecum in response to abrupt inclusion of varying levels of dietary starch. Seven cecally cannulated Quarter Horse geldings (497 to 580 kg) were used in a crossover design with two 28-d periods and a 28-d washout between each. Horses were randomly assigned to dietary treatments consisting of a commercial concentrate offered as fed at either 0.6 (low starch [LS]) or 1.2% BW (high starch [HS]) daily that was divided into 2 meals at 12-h intervals. Prior to the start of each period, horses were allowed ad libitum access to coastal bermudagrass () hay. Concentrate was fed on d 1 with no adaptation. Cecal fluid was collected on d 1 at h 0 and at 3, 6, 9, and 12 h relative to the initial concentrate meal on d 1. Additional samples were collected 6 h after feeding on d 2, 3, and 7 of each period. Cecal contents were used to determine pH and VFA concentrations and extract microbial DNA. The V4 through V6 region of 16S rRNA gene was amplified using PCR and sequenced on the Roche 454 FLX platform. Sequence analysis was performed with QIIME, and data were analyzed using the MIXED procedure of SAS. Cecal pH tended to decrease ( = 0.09) in horses fed HS in the first 12 h after the first concentrate meal and remained lower ( ≤ 0.05) the following 7 d. Total VFA were greater ( ≤ 0.05) in horses fed HS in the initial 12 h and 7 d after addition of concentrate. Species richness determined using the Chao1 index was unchanged ( > 0.20) over the initial 12 h and decreased ( = 0.01) over 7 d for both treatments. Community diversity determined using the Shannon index tended to decrease ( = 0.06) over the 7 d. Relative abundances of Paraprevotellaceae were greater ( ≤ 0.05) in HS in the first 12 h. Over 7 d, relative abundances of Paraprevotellaceae, Veillonellaceae, and Succinivibrionaceae were greater ( ≤ 0.05) in HS compared with LS. Abrupt and short-term exposure to dietary starch does alter cecal fermentation and microbial community structure in horses, but the impact on horse health is unknown.

Influence of oral glucosamine supplementation in young horses challenged with intra-articular lipopolysaccharide.

Fourteen yearling Quarter horses (351 to 470 kg) were utilized in a randomized complete block design to evaluate potential of glucosamine hydrochloride (HCl) to mitigate intra-articular inflammation following a single inflammatory insult. Horses were blocked by BW, age, and sex, and randomly assigned to treatments for a 98-d experiment. Treatments consisted of a control diet (CON; = 7) fed 1% BW per d (as-fed) of concentrate only or a treatment diet ( = 7) of concentrate top dressed with 30 mg/kg BW glucosamine HCl (99.6% purity; GLU30) offered at 12 h intervals. Horses were maintained in individual stalls and offered approximately 1% BW per d of coastal bermudagrass hay (). Plasma and synovial fluid samples were obtained every 14 and 28 d, respectively, and stored at -20°C, before analysis of glucosamine via HPLC. On d 84, an intra-articular lipopolysaccharide (LPS) challenge was conducted on all horses to determine ability of dietary glucosamine HCl supplementation to mitigate joint inflammation and cartilage metabolism. Carpal joints were randomly selected to receive 1 of 2 intra-articular treatments and included sterile lactated Ringer's (control; Contra) only or 0.5 ng LPS solution (LPS) obtained from O55:B5 into the radial carpal joint. Synovial fluid was obtained at pre-injection h 0 and 6, 12, 24, 128, and 336 h post-injection, and was analyzed for prostaglandin E (PGE), carboxypeptide of type II collagen (CPII) and collagenase cleavage neopeptide (C2C) biomarkers by commercial ELISA kits. Data were analyzed using PROC MIXED procedure of SAS. Plasma and synovial glucosamine tended ( = 0.10 and = 0.06, respectively) to increase over time in response to GLU30 compared to CON. There was a treatment by time interaction ( ≤ 0.01), with GLU30 increasing plasma glucosamine concentrations at 28 and 42 d when compared to CON. A treatment by time interaction ( ≤ 0.01) was observed with GLU30 increasing synovial glucosamine levels at d 28 and 84 ( ≤ 0.01 and = 0.05, respectively). Intra-articular LPS increased ( ≤ 0.01) synovial PGE, C2C, and CPII levels. GLU30 decreased synovial PGE and C2C concentrations when compared to CON ( = 0.04 and = 0.05, respectively), while synovial levels of CPII increased ( ≤ 0.01) in GLU30 horses. These results indicate the potential for oral glucosamine HCl to mitigate intra-articular inflammation and influence cartilage turnover in a young horse model.

Gastric adenocarcinoma associated with human immunodeficiency virus (HIV) infection.

Gastric adenocarcinoma developed in a 33-year-old homosexual man with human immunodeficiency virus (HIV) infection. This type of cancer is very rare in a man of his young age and in the absence of other factors. The tumor was aggressive and led to rapid death. Whereas Kaposi's sarcoma and lymphomas have been associated with HIV, there are only a few documented cases of adenocarcinoma associated with a suppressed immune system. The association between HIV infection and the virulence of this tumor appears to be strong.