Joint work and power for both the forelimb and hindlimb during trotting in the horse.

The net work of the limbs during constant speed over level ground should be zero. However, the partitioning of negative and positive work between the fore- and hindlimbs of a quadruped is not likely to be equal because the forelimb produces a net braking force while the hindlimb produces a net propulsive force. It was hypothesized that the forelimb would do net negative work while the hindlimb did net positive work during trotting in the horse. Because vertical and horizontal impulses remain unchanged across speeds it was hypothesized that net work of both limbs would be independent of speed. Additionally because the major mass of limb musculature is located proximally, it was hypothesized that proximal joints would do more work than distal joints. Kinetic and kinematic analysis were combined using inverse dynamics to calculate work and power for each joint of horses trotting at between 2.5 and 5.0 m s(-1). Work done by the hindlimb was indeed positive (consistently 0.34 J kg(-1) across all speeds), but, contrary to our hypothesis, net work by the forelimb was essentially zero (but also independent of trotting speed). The zero net work of the forelimb may be the consequence of our not being able to account, experimentally, for the negative work done by the extrinsic muscles connecting the scapula and the thorax. The distal three joints of both limbs behaved elastically with a period of energy absorption followed by energy return. Proximal forelimb joints (elbow and shoulder) did no net work, because there was very little movement of the elbow and shoulder during the portion of stance when an extensor moment was greatest. Of the two proximal hindlimb joints, the hip did positive work during the stride, generating energy almost throughout stance. The knee did some work, but like the forelimb proximal joints, had little movement during the middle of stance when the flexion moment was the greatest, probably serving to allow the efficient transmission of energy from the hip musculature to the ground.

Additional food folate derived exclusively from natural sources improves folate status in young women with the MTHFR 677 CC or TT genotype.

The effectiveness of additional food folate in improving folate status in humans is uncertain particularly in people with the common genetic variant (677 C-->T) in the methylenetetrahydrofolate reductase (MTHFR) gene. To examine the effect of a doubling of food folate consumption on folate status response variables, women (n=32; 18-46 years) with the MTHFR 677 CC or TT genotype consumed either 400 (n=15; 7 CC and 8 TT) or 800 (n=17; 8 CC and 9 TT) microg/day of dietary folate equivalents (DFE) derived exclusively from naturally occurring food folate for 12 weeks. A repeated measures two-factor ANOVA was used to examine the effect of the dietary treatment, the MTHFR C677T genotype and their interactions on serum folate, RBC folate and plasma total homocysteine (tHcy) during the last 3 weeks of the study. Consumption of 800 microg DFE/day resulted in serum folate concentrations that were 67% (P=.005) higher than consumption of 400 microg DFE/day (18.6+/-2.9 vs. 31.0+/-2.7 nmol/L, respectively) and RBC folate concentrations that were 33% (P=.001) higher (1172+/-75 vs. 1559+/-70 nmol/L, respectively). Serum folate (P=.065) and RBC folate (P=.022) concentrations were lower and plasma tHcy was higher (P=.039) in women with the MTHFR 677 TT genotype relative to the CC genotype. However, no genotype by dietary treatment interaction was detected. These data suggest that a doubling of food folate intake will lead to marked improvements in folate status in women with the MTHFR 677 CC or TT genotype.

The glycine N-methyltransferase (GNMT) 1289 C->T variant influences plasma total homocysteine concentrations in young women after restricting folate intake.

Glycine N-methyltransferase (GNMT) is a key regulatory protein in folate metabolism, methionine availability, and transmethylation reactions. Perturbations in GNMT may lead to aberrations in homocysteine metabolism, a marker of numerous pathologies. The primary objective of this study was to examine the influence of the GNMT 1289 C-->T alone, and in combination with the methylenetetrahydrofolate reductase (MTHFR) 677 C-->T variant, on plasma total homocysteine concentrations in healthy young women (n = 114). Plasma total homocysteine was measured at baseline (wk 0) and after 2 wk of controlled folate restriction (135 microg/d as dietary folate equivalents). Plasma homocysteine concentrations did not differ among the GNMT C1289T genotypes at baseline. However, after folate restriction, women with the GNMT 1289 TT genotype (n = 16) had higher (P = 0.019) homocysteine concentrations than women with the CT (n = 51) or CC (n = 47) genotype. The influence of the GNMT 1289 C-->T variant on homocysteine was dependent on the MTHFR C677T genotype. In subjects with the MTHFR 677 CC genotype, homocysteine was greater (P < or = 0.05) for GNMT 1289 TT subjects relative to 1289 CT or CC subjects. However, in subjects with the MTHFR 677 TT genotype, plasma homocysteine concentrations did not differ among the GNMT C1289T genotypes. Overall, these data suggest that the GNMT 1289 C-->T polymorphism influences plasma homocysteine and is responsive to folate intake.

A long-term controlled folate feeding study in young women supports the validity of the 1.7 multiplier in the dietary folate equivalency equation.

The presence of folic acid in enriched cereal grain products and the higher bioavailability of folic acid than food folate led to the expression of the 1998 folate RDA, 400 microg/d, as dietary folate equivalents (DFE). DFE are defined as: mug natural food folate + 1.7 x microg synthetic folic acid. The 1.7 multiplier was based on assumptions that added folic acid was 85% available and food folate was 50% available. The 85/50 ratio also inferred that the bioavailability of food folate was approximately 60% relative to added folic acid. The objective of this long-term controlled feeding study was to assess the dietary folate equivalency of folic acid. After a 2-wk period of folate restriction, women (n = 42, 18-45 y old) consumed either 400 or 800 microg DFE/d derived from various combinations of food folate and folic acid for 12 wk. Folic acid was converted to DFE using the 1.7 multiplier from the DFE calculation and was consumed with a meal throughout the treatment period. Folate status response to the various treatments was assessed during wk 12-14. Serum folate, RBC folate, and plasma total homocysteine did not differ among the 400 microg DFE/d groups or among the 800 microg DFE/d groups. In contrast, consumption of 800 microg DFE/d led to higher (P

In vivo muscle function vs speed. II. Muscle function trotting up an incline.

Different locomotor tasks, such as moving up or down grades or changing speed, require that muscles adjust the amount of work they perform to raise or lower, accelerate or decelerate the animal's center of mass. During level trotting in the horse, the triceps had shortening strains of around 10.6% while the vastus shortened 8.1% during the stance phase. Because of the 250% increase in metabolic rate in horses trotting up a 10% incline which is, presumably, a result of the increased requirement for mechanical work, we hypothesized that muscle strain during trotting would be increased in both the triceps and the vastus over that observed when trotting on the level. Because times of contact are similar in level and incline trotting, we also hypothesized that strain rates of these muscles would be increased, accompanied by an increase in EMG activity. We examined the lateral head of the triceps and the vastus lateralis while trotting up a 10% incline (5.7 degrees) over a range of speeds. The triceps shortened by 18% compared with 10.6% shortening on the level, and the vastus shortened by 18.5% compared with 8.1% on the level. The increased shortening velocities that were observed in both muscles probably reduced the force that any given set of activated muscle fibers could produce. If this pattern held for other limb muscles that do work to elevate the horse's center of mass on an incline, then a greater volume of muscle would have to be recruited to generate an equivalent force for body support. This was reflected in significant increases in the EMG intensity (IEMG) of both muscles.

In vivo muscle function vs speed. I. Muscle strain in relation to length change of the muscle-tendon unit.

The activity of muscles can be concentric (shortening), eccentric (lengthening) or isometric (constant length). When studying muscle function it is important to know what the muscle fascicles are actually doing because the performance of muscle is strongly influenced by the type of activity: force decreases as a function of shortening velocity during concentric contractions; force produced during eccentric contractions can be stronger than maximum isometric force, and force production is enhanced if a concentric contraction follows an eccentric phase. It is well known that length changes of muscle fascicles may be different from length changes of the overall muscle-tendon unit because of the compliance of the series elasticity. Consequently, fascicles of joint extensor muscles may not undergo eccentric activity even when the joint flexes, but the extent to which this occurs may vary with the compliance of the series elasticity and may differ between species: the vastus lateralis, a knee extensor, shortens when active during trotting in dogs and lengthens in rats. Previous studies of kinematics of trotting in horses have shown that during stance, the elbow extends nearly continuously with a brief period of flexion near mid-stance and the knee exhibits two phases of flexion followed by extension. The lateral triceps (an elbow extensor) has no external tendon but the vastus lateralis has a relatively long external tendon and the fascicles insert on an aponeurosis. Thus, one might expect the relation between fascicle strain and overall length change of the muscle-tendon units to be quite different in these two muscles. In the present study in horses, fascicle length changes of the lateral triceps and vastus lateralis were measured with sonomicrometry and length changes of the muscle-tendon units were estimated from muscle architecture and joint kinematics for four horses trotting on a treadmill at nine speeds. Because the focus of this study was the relation between length changes of the muscle-tendon unit (estimated from kinematics) and length changes in the muscle fascicles, we divided the stance-phase sonomicrometry records into phases that corresponded to the alternating flexion and extension of the joint as indicated by the kinematic records. During its one eccentric phase, the triceps shortened by 0.7+/-0.4% despite a predicted lengthening of 1%. Similarly, the vastus shortened by 3.7+/-1.9% when kinematics predicted 3.2% lengthening. During their concentric phases the triceps shortened by 10.6% and the vastus shortened by 8.1%. Strain in the triceps did not change with speed but it did in the vastus. Strain rate increased with speed in both muscles as did the integrated EMG, indicating an increase in the volume of muscle recruited. Thus, despite differences in their architecture and the kinematic patterns of the associated joints, these two joint extensors exhibited similar activity.

Ground reaction forces in horses trotting up an incline and on the level over a range of speeds.

Although the forces required to support the body mass are not elevated when moving up an incline, kinematic studies, in vivo tendon and bone studies and kinetic studies suggest there is a shift in forces from the fore- to the hindlimbs in quadrupeds. However, there are no whole-animal kinetic measurements of incline locomotion. Based on previous related research, we hypothesized that there would be a shift in forces to the hindlimb. The present study measured the force produced by the fore- and hindlimbs of horses while trotting over a range of speeds (2.5 to 5 m s(-1)) on both level and up an inclined (10%) surface. On the level, forelimb peak forces increased with trotting speed, but hindlimb peak force remained constant. On the incline, both fore- and hindlimb peak forces increased with speed, but the sum of the peak forces was lower than on the level. On the level, over the range of speeds tested, total force was consistently distributed between the limbs as 57% forelimb and 43% hindlimb, similar to the weight distribution of the horses during static weight tests. On the incline, the force distribution during locomotion shifted to 52% forelimb and 48% hindlimb. Time of contact and duty factor decreased with speed for both limbs. Time of contact was longer for the forelimb than the hindlimb, a finding not previously reported for quadrupeds. Time of contact of both limbs tended to be longer when traveling up the incline than on the level, but duty factor for both limbs was similar under both conditions. Duty factor decreased slightly with increased speed for the hindlimb on the level, and the corresponding small, predicted increase in peak vertical force could not be detected statistically.

Moments and power generated by the horse (Equus caballus) hind limb during jumping.

The ability to jump over an obstacle depends upon the generation of work across the joints of the propelling limb(s). The total work generated by one hind limb of a horse and the contribution to the total work by four joints of the hind limb were determined for a jump. It was hypothesized that the hip and ankle joints would have extensor moments performing positive work, while the knee would have a flexor moment and perform negative work during the jump. Ground reaction forces and sagittal plane kinematics were simultaneously recorded during each jumping trial. Joint moment, power and work were determined for the metatarsophalangeal (MP), tarsal (ankle), tibiofemoral (knee) and coxofemoral (hip) joints. The hip, knee and ankle all flexed and then extended and the MP extended and then flexed during ground contact. Consistent with our hypothesis, large extensor moments were observed at the hip and ankle joints and large flexor moments at the knee and MP joints throughout ground contact of the hind limb. Peak moments tended to occur earlier in stance in the proximal joints but peak power generation of the hind limb joints occurred at similar times except for the MP joint, with the hip and ankle peaking first followed by the MP joint. During the first portion of ground contact (approximately 40%), the net result of the joint powers was the absorption of power. During the remainder of the contact period, the hind limb generated power. This pattern of power absorption followed by power generation paralleled the power profiles of the hip, ankle and MP joints. The total work performed by one hind limb was 0.71 J kg(-1). Surprisingly, the knee produced 85% of the work (0.60 J kg(-1)) done by the hind limb, and the positive work performed by the knee occurred during the first 40% of the take-off. There is little net work generated by the other three joints over the entire take-off. Velocity of the tuber coxae (a landmark on the pelvis of the animal) was negative (downward) during the first 40% of stance, which perhaps reflects the negative work performed to decrease the potential energy during the first 40% of contact. During the final 60% of contact, the hip, ankle and MP joints generate positive work, which is reflected in the increase of the animal's potential energy.

Methylenetetrahydrofolate reductase 677C-->T variant modulates folate status response to controlled folate intakes in young women.

A common genetic variant in the methylenetetrahydrofolate reductase (MTHFR) gene involving a cytosine to thymidine (C-->T) transition at nucleotide 677 is associated with reduced enzyme activity, altered folate status and potentially higher folate requirements. The objectives of this study were to investigate the effect of the MTHFR 677 T allele on folate status variables in Mexican women (n = 43; 18-45 y) and to assess the adequacy of the 1998 folate U.S. Recommended Dietary Allowance (RDA), 400 micro g/d as dietary folate equivalents (DFE). Subjects (14 CC, 12 CT, 17 TT genotypes) consumed a low folate diet (135 micro g/d DFE) for 7 wk followed by repletion with 400 micro g/d DFE (7 CC, 6 CT, 9 TT) or 800 micro g/d DFE (7 CC, 6 CT, 8 TT) for 7 wk. Throughout repletion with 400 micro g/d DFE, the TT genotype had lower (P 0.05) in their response relative to the CC genotype. Throughout repletion with 800 micro g/d DFE, the CT genotype had lower (P 0.05) in the measured variables between the TT and CC genotypes. Repletion with 400 micro g/d DFE led to normal blood folate and desirable plasma tHcy concentrations, regardless of MTHFR C677T genotype. Collectively, these data demonstrate that the MTHFR C-->T variant modulates folate status response to controlled folate intakes and support the adequacy of the 1998 folate U.S. RDA for all three MTHFR C677T genotypes.

The energetics of the trot-gallop transition.

Two studies have focused on potential triggers for the trot-gallop transition in the horse. One study concluded that the transition was triggered by metabolic economy. The second study found that it was not metabolic factors but, rather, peak musculoskeletal forces that determine gait transition speeds. In theory, peak musculoskeletal forces should be the same when trotting up an incline as when trotting at the same speed on the level. Assuming this is the case, we hypothesized that if peak forces determine gait transition speeds then horses should switch from a trot to a gallop at the same speed (i.e. the same critical force) regardless of incline. The aim of the present research was to examine the effects of incline on the trot-gallop transition speed in horses and to re-examine the role of metabolism in determining the trot-gallop transition. Horses (Equus caballus) were conditioned to run on a high-speed treadmill prior to data collection. Gait changes were recorded for each horse using a standardized testing protocol on the level and when trotting up a 10% incline. Both maximum sustained trotting speeds and minimum sustained galloping speeds, representing the lower and upper limits of the trot-gallop transition, respectively, were significantly slower when trotting up an incline. After completing collection of gait transition data, the horses were trained to extend their gaits beyond the normal transition speeds, and metabolic data were collected. Maximum sustained trotting speeds were not different from the energetically optimal transition speeds, i.e. the speed at which metabolic rates are the same for both gaits.

Heterogeneity in the prevalence of methylenetetrahydrofolate reductase gene polymorphisms in women of different ethnic groups.

OBJECTIVE: To determine the prevalence of methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms in women of different ethnic groups and to relate these common mutations to plasma homocysteine, red cell folate, and serum folate. DESIGN: A one-time fasting blood sample was obtained for MTHFR genotype (C677T and A1298C) determinations (n=433). Serum folate, red cell folate, and homocysteine analyses were performed in nonfolic acid supplement users (n=215). SUBJECTS/SETTING: This study involved 433 women from four ethnic groups, including 193 Hispanic women of Mexican descent, 139 white women, 53 Asian women of mixed descent, and 48 African American women. STATISTICAL ANALYSIS PREFORMED: Chi;(2), t Test, and analysis of variance were used. RESULTS: Mexican women (18.1%) had a higher frequency of the 677 TT genotype compared with white (7.2%), Asian (3.8%), and African American (0%) women. White women (7.9%) had a higher frequency of the 1298 CC genotype than the other ethnic groups (range=1.9% to 2.6%). The frequency of compound heterozygosity (677 CT + 1298 AC) was higher in Mexican (17.6%) and white (15.1%) women than Asian and African American ( approximately 4% to 6%) women. In the era of folic acid fortification, neither genotype, independently or together, was associated with homocysteine or blood folate concentrations when ethnic groups were combined. In Mexican women, however, a linear trend (P