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The physiological cost of walking is greater in bilateral amputees (BA) than in both unilateral amputee and non-pathological gait. The aim of this study was to describe the physiological costs and other standard gait characteristics in a sample population of BA, walking at self-selected (comfortable) speeds. Amputees had bilateral trans-tibial, bilateral trans-femoral or trans-tibial/trans-femoral amputations as a result of trauma or congenital defects. All amputees wore their own prosthetic limbs which were either full-length prostheses or short non-articulating pylon prostheses (SNAPPs). The results were compared with a base line data set collected from a non-pathological control group. It was anticipated that amputees with high-level amputations would walk at the slowest speeds, have the highest physiological costs and lowest perception of walking ability. However, varying walking speeds resulted in varying exercise intensities, exercise heart rates and perceptions of walking that could not be directly related to amputation levels. It is therefore concluded that bilateral amputee gait is complex, varied and not easily categorized. [ABSTRACT FROM AUTHOR]

Tomkins et al. [A. G. Tomkins et al. , Nature 533, 235-238 (2016)] suggested that iron oxides contained in 2.7-Ga iron micrometeorites can be used to determine the concentration of O 2 in the Archean upper atmosphere. Specifically, they argued that the presence of magnetite in these objects implies that O 2 must have been near present-day levels (∼21%) within the altitude range where the micrometeorites were melted during entry. Here, we reevaluate their data using a 1D photochemical model. We find that atomic oxygen, O, is the most abundant strong oxidant in the upper atmosphere, rather than O 2 But data from shock tube experiments suggest that CO 2 itself may also serve as the oxidant, in which case micrometeorite oxidation really constrains the CO 2 /N 2 ratio, not the total oxidant abundance. For an atmosphere containing 0.8 bar of N 2 , like today, the lower limit on the CO 2 mixing ratio is ∼0.23. This would produce a mean surface temperature of ∼300 K at 2.7 Ga, which may be too high, given evidence for glaciation at roughly this time. If p N 2 was half the present value, and warming by other greenhouse gases like methane was not a major factor, the mean surface temperature would drop to ∼291 K, consistent with glaciation. This suggests that surface pressure in the Neoarchean may need to have been lower-closer to 0.6 bar-for CO 2 to have oxidized the micrometeorites. Ultimately, iron micrometeorites may be an indicator for ancient atmospheric CO 2 and surface pressure; and could help resolve discrepancies between climate models and existing CO 2 proxies such as paleosols., Competing Interests: The authors declare no competing interest.

Selective breeding for performance has resulted in distinct breeds of horse, such as the Quarter Horse (bred for acceleration) and the Arab (bred for endurance). Rapid acceleration, seen during Quarter Horse racing, requires fast powerful muscular contraction and the generation of large joint torques, particularly by the hind limb muscles. This study compared hind limb moment arm lengths in the Quarter Horse and Arab. We hypothesized that Quarter Horse hind limb extensor muscles would have longer moment arms when compared to the Arab, conferring a greater potential for torque generation at the hip, stifle and tarsus during limb extension. Six Quarter Horse and six Arab hind limbs were dissected to determine muscle moment arm lengths for the following muscles: gluteus medius, biceps femoris, semitendinosus, vastus lateralis, gastrocnemius (medialis and lateralis) and tibialis cranialis. The moment arms of biceps femoris (acting at the hip) and gastrocnemius lateralis (acting at the stifle) were significantly longer in the Quarter Horse, although the length of the remaining muscle moment arms were similar in both breeds of horse. All the Quarter Horse muscles were capable of generating greater muscle moments owing to their greater physiological cross-sectional area (PCSA) and therefore greater isometric force potential, which suggests that PCSA is a better determinant of muscle torque than moment arm length in these two breeds of horse. With the exception of gastrocnemius and tibialis cranialis, the observed muscle fascicle length to moment arm ratio (MFL : MA ratio) was greater for the Arab horse muscles. It appears that the Arab muscles have the potential to operate at slower velocities of contraction and hence generate greater force outputs when compared to the Quarter Horse muscles working over a similar range of joint motion; this would indicate that Arab hind limb muscles are optimized to function at maximum economy rather than maximum power output.

We provide quantitative anatomical data on the muscle-tendon architecture and geometry of the pelvic limb of an elite sprint athlete, the racing greyhound. Specifically, muscle masses, muscle lengths, fascicle lengths, pennation angles and muscle moment arms were measured. Maximum isometric force and power of muscles, the maximum muscle torque at joints and tendon stress and strain were estimated. We compare data with that published for a generalized breed of canid, and other cursorial mammals such as the horse and hare. The pelvic limb of the racing greyhound had a relatively large volume of hip extensor muscle, which is likely to be required for power production. Per unit body mass, some pelvic limb muscles were relatively larger than those in less specialized canines, and many hip extensor muscles had longer fascicle lengths. It was estimated that substantial extensor moments could be created about the tarsus and hip of the greyhound allowing high power output and potential for rapid acceleration. The racing greyhound hence possesses substantial specializations for enhanced sprint performance.

We provide quantitative muscle-tendon architecture and geometry data for the racing greyhound thoracic limb. Muscle mass, belly length, fascicle lengths, pennation angles and moment arms were measured, as were tendon masses and lengths. Maximum isometric force and maximum power were estimated for muscles, and maximum stress and strain were estimated for tendons. Results are compared with other fast quadrupedal runners, and to previously published data in mixed-breed dogs. The implications of the functional adaptations of the greyhound thoracic limb for sprinting performance are discussed. The thoracic limb was found to benefit from a similar proportion of locomotor muscle mass to the pelvic limb, suggesting that it may be used to some extent in propulsion, or alternatively that stabilisation is very important in this animal. Extrinsic muscles, especially latissimus dorsi and pectoralis profundus, were predicted to be powerful and important for generating net positive work during accelerations. Proximal biarticular muscles show specialisation toward preventing collapse of the shoulder and elbow joints to enable strut-like limb function, or some form of dynamic control. Distal muscles did not appear specialised for elastic energy storage, a functional difference to pelvic limb muscles, and the equivalents in horse thoracic limbs. The greyhound thoracic limb appears to possess substantial differences from both that of more 'sub-maximal specialist' quadrupeds, and from the greyhound pelvic limb.

The evolution of habitual bipedalism is still a fundamental yet unsolved question for paleoanthropologists, and carrying is popular as an explanation for both the early adoption of upright walking and as a positive selection pressure once a terrestrial lifestyle had been adopted. However, to support or reject any hypothesis that suggests carrying efficiency was an important selective pressure, we need quantitative data on the costs of different forms of carrying behavior, especially infant-carrying since reduction in the grasping capabilities of the foot would have prevented infants from clinging on for long durations. In this study, we tested the hypothesis that the mode of load carriage influences the energetic cost of locomotion. Oxygen consumption was measured in seven female participants walking at a constant speed while carrying four different 10-kg loads (a weighted vest, 5-kg dumbbells carried in each hand, a mannequin infant carried on one hip, and a 10-kg dumbbell carried in a single hand). Oxygen consumption was also measured during unloaded standing and unloaded walking. The results show that the weighted vest requires the least amount of energy of the four types of carrying and that, for this condition, humans are as efficient as mammals in general. The balanced load was carried with approximately the predicted energy cost. However, the asymmetrical conditions were considerably less efficient, indicating that, unless infant-carrying was the adaptive response to a strong environmental selection pressure, this behavior is unlikely to have been the precursor to the evolution of bipedalism.

The Quarter Horse (bred for acceleration) and the Arab (bred for endurance) are situated at either end of the equine athletic spectrum. Studies into the form and function of the leg muscles in human sprint and endurance runners have demonstrated that differences exist in their muscle architecture. It is not known whether similar differences exist in the horse. Six Quarter Horse and six Arab fresh hind limb cadavers were dissected to gain information on the muscle mass and architecture of the following muscles: gluteus medius; biceps femoris; semitendinosus; vastus lateralis; gastrocnemius; tibialis cranialis and extensor digitorum longus. Specifically, muscle mass, fascicle length and pennation angle were quantified and physiological cross-sectional area (PCSA) and maximum isometric force were estimated. The hind limb muscles of the Quarter Horse were of a significantly greater mass, but had similar fascicle lengths and pennation angles when compared with those of the Arab; this resulted in the Quarter Horse hind limb muscles having greater PCSAs and hence greater isometric force potential. This study suggests that Quarter Horses as a breed inherently possess large strong hind limb muscles, with the potential to accelerate their body mass more rapidly than those of the Arab.

Muscle moment arms were measured for major muscles of the pelvic limb of the ostrich (Struthio camelus) in order to assess specific functional behaviour and to apply this to locomotor performance. Pelvic limbs of six juvenile ostriches were used for this study. The tendon travel technique was used to measure moment arms of 21 muscles at the hip, knee, ankle and metatarsophalangeal joints throughout the ranges of motion observed during level running. Six of the 21 muscles measured were found to have moment arms that did not change with joint angle, whilst the remainder all demonstrated angle-dependent changes for at least one of the joints crossed. Moment arm lengths tended to be longest for the large proximal muscles, whilst the largest relative changes were found for the moment arms of the distal muscles. For muscles where moment arm varied with joint angle: all hip muscles were found to have increasing moment arms with extension of the joint, knee flexors were found to have moment arms that increased with extension, knee extensor moment arms were found to increase with flexion and ankle extensor moment arms increased with extension. The greatest relative changes were observed in the flexors of the metatarsophalangeal joint, for which a three-fold increase in moment arm was observed from flexion to full extension. Changes in muscle moment arm through the range of motion studied appear to optimize muscle function during stance phase, increasing the effective mechanical advantage of these muscles.

We provide quantitative anatomical data on the muscle-tendon architecture of the hare thoracic limb (specifically muscle mass, fascicle length, pennation angle, tendon mass and length). In addition, moment arms of major thoracic limb muscles were measured. Maximum isometric force and power of muscles, the moment of force about a joint, and tendon stress and strain were estimated. Data are compared with those from other cursorial mammals. The thoracic limb of the hare consists predominantly of extrinsic musculature with long parallel fascicles, specialised for generating force over a large range. A large shoulder flexor/elbow extensor muscle mass is present, in particular Triceps brachii. The pennate nature of the long head of this muscle suggests it has an important role in stabilising the elbow joint during stance, whilst moment arm curves suggest that it may also play a role in initiating shoulder flexion. In addition, Supraspinatus and Infraspinatus are capable of generating high forces, potentially to stabilise the shoulder joint during the stance phase of locomotion. Supraspinatus may in addition play an important role in forelimb protraction. The Subscapularis muscle was capable of generating surprisingly high forces, suggesting that the hare must be able to withstand/produce high forces during activities that need medio-lateral stability, such as turning. Distally, tendons were relatively short, showing little potential for elastic energy storage when compared with both their pelvic limb counterparts and their equivalents in the horse thoracic limb. Thus, a 'stiffer' thoracic limb may be beneficial in terms of behaving like a strut, simply supporting and deflecting the body during high-speed running. This more distal/less proximal distribution of limb mass is also likely to be important in retaining the manipulative/adaptive/non-locomotor capabilities of the limb.

We provide quantitative anatomical data on the muscle-tendon architecture of the hare pelvic limb (specifically muscle mass, fascicle length, pennation angle, tendon mass and length). In addition, moment arms of major pelvic limb muscles were measured. Maximum isometric force and power of muscles, the moment of force about a joint, and tendon stress and strain were estimated. Data are compared with published data for other cursorial mammals such as the horse and dog, and a non-specialised Lagamorph, the rabbit. The pelvic limb of the hare was found to contain substantial amounts of hip extensor and adductor/abductor muscle volume, which is likely to be required for power production and stability during rapid turning. A proximal to distal reduction in muscle volume and fascicle length was also observed, as is the case in other cursorial quadrupeds, along with a reduction in distal limb mass via the replacement of muscle volume by long distal limb tendons, capable of storing large amounts of elastic energy. The majority of hare pelvic limb muscle moment arms varied with joint position, giving the hare the capacity to vary muscle function with limb posture and presumably different locomotor activities.

The functional anatomy of the pelvic limb of the ostrich (Struthio camelus) was investigated in order to assess musculoskeletal specialization related to locomotor performance. The pelvic limbs of ten ostriches were dissected and detailed measurements of all muscle tendon units of the pelvic limb were made, including muscle mass, muscle length, fascicle length, pennation angle, tendon mass and tendon length. From these measurements other muscle properties such as muscle volume, physiological cross-sectional area (PCSA), tendon cross-sectional area, maximum isometric muscle force and tendon stress were derived, using standard relationships and published muscle data. Larger muscles tended to be located more proximally and had longer fascicle lengths and lower pennation angles. This led to an expected proximal to distal reduction in total muscle mass. An exception to this trend was the gastrocnemius muscle, which was found to have the largest volume and PCSA and also had the highest capacity for both force and power production. Generally high-power muscles were located more proximally in the limb, while some small distal muscles (tibialis cranialis and flexor perforatus digiti III), with short fibres, were found to have very high force generation capacities. The greatest proportion of pelvic muscle volume was for the hip extensors, while the highest capacity for force generation was observed in the extensors of the ankle, many of which were also in series with long tendons and thus were functionally suited to elastic energy storage.

Quantitative, accurate data regarding the inertial properties of body segments are of paramount importance when developing musculo-skeletal locomotor models of living animals and, by inference, their ancestors. The limited number of available primate cadavers, and the destructive nature of the post-mortem, result in such data being very rare for primates. This study builds on the work of Crompton et al. (Am. J. Phys. Anthropol. 1996, 99, 547-570) and reports inertial properties of the body segments of gorillas, chimpanzees, orangutans and gibbons. Segment mass, centre of mass and the radius of gyration of five ape cadavers were measured using a complex-pendulum technique and compared with the results derived from external measurements of segment lengths and diameters on the same animals. With additional data from external measurements of eight more hominoid cadavers, and published data, intergeneric differences between the inertial properties and the distribution of mass between limb segments are analysed and related to the locomotor habits of the species. We found that segment inertial properties show extensive overlap between ape genera as a result of large interindividual variation. Segment mass distribution also overlaps between apes and humans, with the exception of the shank segment. However, owing to a different distribution of mass between the limb segments, the centre of mass of both the arms and the legs is located more distally in apes than in humans, and the natural pendular period of ape forelimbs is larger than that of the hindlimbs. This suggests that, in contrast to the limbs of cursorial mammals and cercopithecoid primates, hominoid limbs are not optimized for efficiency in quadrupedal walking, but rather reflect a compromise between various locomotor modes. Common chimpanzees may have secondarily evolved a more efficient quadrupedal gait.

Flexion/extension moment arms were obtained for the major muscles crossing the hip, knee and ankle joints in the orang-utan, gibbon, gorilla (Eastern and Western lowland) and bonobo. Moment arms varied with joint motion and were generally longer in proximal limb muscles than distal limb muscles. The shape of the moment arm curves (i.e. the plots of moment arm against joint angle) differed in different hindlimb muscles and in the same muscle in different subjects (both in the same and in different ape species). Most moment arms increased with increasing joint flexion, a finding which may be understood in the context of the employment of flexed postures by most non-human apes (except orang-utans) during both terrestrial and arboreal locomotion. When compared with humans, non-human great apes tended to have muscles better designed for moving the joints through large ranges. This was particularly true of the pedal digital flexors in orang-utans. In gibbons, the only lesser ape studied here, many of the moment arms measured were relatively short compared with those of great apes. This study was performed on a small sample of apes and thus differences noted here warrant further investigation in larger populations.

The purpose of this work was to evaluate the accuracy and precision of a freehand three-dimensional (3-D) ultrasonography system in the determination of lesion volume in tendons. The accuracy and precision of a 3-D ultrasonography system was assessed by performing repeated measurements on a phantom of known volume. Volume measurements of tendon lesions performed with 3-D ultrasonography were compared with measurements based on a series of two-dimensional (2-D) ultrasound (US) scans and to direct measurements from dissections. A novel method for the creation of tendon lesions in vitro was developed. 3-D US showed excellent precision and accuracy in measurements of the phantom (mean measured volume = 3.76 mL, calculated volume = 3.77 mL, coefficient of variation (CoV) = 0.54%) and good repeatability in the determination of tendon lesions (repeatability coefficient = 0.00047). All three methods examined were repeatable (repeatability coefficient for 2-D US = 0.00032, repeatability coefficient for dissections = 0.00076). However, each of the methods produced different results and no constant relationship could be found between any of the measurement methods. Both 3-D and 2-D US proved to be repeatable techniques for the measurement of the volume of a tendon lesion. Even if they produced different results, each of them can be repeatedly used individually. It was not possible to define which one provided the most accurate value as a result of difficulties encountered in lesion identification on histology, and therefore the lack of a gold standard.

We provide quantitative anatomical data on the muscle-tendon units of the equine pelvic limb. Specifically, we recorded muscle mass, fascicle length, pennation angle, tendon mass and tendon rest length. Physiological cross sectional area was then determined and maximum isometric force estimated. There was proximal-to-distal reduction in muscle volume and fascicle length. Proximal limb tendons were few and, where present, were relatively short. By contrast, distal limb tendons were numerous and long in comparison to mean muscle fascicle length, increasing potential for elastic energy storage. When compared with published data on thoracic limb muscles, proximal pelvic limb muscles were larger in volume and had shorter fascicles. Distal limb muscle architecture was similar in thoracic and pelvic limbs with the exception of flexor digitorum lateralis (lateral head of the deep digital flexor), the architecture of which was similar to that of the pelvic and thoracic limb superficial digital flexors, suggesting a functional similarity.

Muscles have two major roles in locomotion: to generate force and to absorb/generate power (do work). Economical force generation is achieved by short-fibred pennate muscle while the maximum power output of a muscle is architecture independent. In this study we tested the hypothesis that there is an anatomical and structural separation between the force-generating anti-gravity muscles and the propulsive (limb/trunk moving) muscles of the equine forelimb. Muscle mass and fascicle length measurements were made on the thoracic limb extrinsic muscles of six fresh horse cadavers. Physiological cross-sectional area and maximum isometric force were then estimated. Maximum power was estimated from muscle volume and published contraction velocity data. The majority of extrinsic forelimb muscles were large with long fascicles arranged in parallel to the long axis of the muscle. Muscles arranged in this way are optimised for doing work. The architecture of serratus ventralis thoracis (SVT) was unique. It had short (48 +/- 17 mm) fascicles, arranged at about 45 degrees to the long axis of the muscle, which would suggest a force-generating, anti-gravity role. The muscle belly of SVT was sandwiched between two broad, thick sheets of aponeurosis. Hence, SVT could make a significant contribution to the overall elastic properties of the thoracic limb.

Muscles have two major roles in locomotion: to generate force and to absorb/generate power (do work). Economical force generation is achieved by short-fibred pennate muscle while the maximum power output of a muscle is architecture independent. In this study we tested the hypothesis that there is an anatomical and structural separation between the force-generating anti-gravity muscles and the propulsive (limb/trunk moving) muscles of the equine forelimb. Muscle mass and fascicle length measurements were made on the thoracic limb extrinsic muscles of six fresh horse cadavers. Physiological cross-sectional area and maximum isometric force were then estimated. Maximum power was estimated from muscle volume and published contraction velocity data. The majority of extrinsic forelimb muscles were large with long fascicles arranged in parallel to the long axis of the muscle. Muscles arranged in this way are optimised for doing work. The architecture of serratus ventralis thoracis (SVT) was unique. It had short (48 +/- 17 mm) fascicles, arranged at about 45 degrees to the long axis of the muscle, which would suggest a force-generating, anti-gravity role. The muscle belly of SVT was sandwiched between two broad, thick sheets of aponeurosis. Hence, SVT could make a significant contribution to the overall elastic properties of the thoracic limb.

Background: The extracellular domain of the HER-2/neu oncogene product is increased in sera of some patients with epithelial cancers. Our aim was to develop an automated serum assay for the extracellular domain of the HER-2/neu protein., Methods: We used a monoclonal antibody labeled with fluorescein for capture and a monoclonal Fab' fragment labeled with alkaline phosphatase for detection. Separation of bound and free detection conjugate was performed with magnetizable particles coated with monoclonal antibody to fluorescein. Alkaline phosphatase activity was measured kinetically at 405 or 450 nm., Results: The assay was linear from 0.1 to 250 microg/L. No hook effect was evident up to 10 000 microg/L. Within-run imprecision (CV) was 0.8-1.2%, and total imprecision was 1.1-1.7%. Cross-reactivity with human epidermal growth factor receptor, which has extensive homology with HER-2/neu extracellular domain, was <0.6%. Human anti-mouse antibodies, heterophilic antibodies, and rheumatoid factor did not interfere, nor did the therapeutic monoclonal antibody Herceptin((R)). In 51 healthy females, the mean value was 9.3 microg/L with a range of 6.4-14.0 microg/L. No reagent lot-to-lot variability was detected over four lots of reagents tested., Conclusion: The Bayer Immuno 1(TM) assay for HER-2/neu was precise and resistant to interferences, characteristics that are essential for longitudinal monitoring of cancer patients.

Two studies on the efficacy of mel Cy (Cymelarsan, Rhone Merieux, France) for the treatment of cattle infected with Trypanosoma evansi were carried out with groups of 5 Friesian Holstein calves infected with an Indonesian stock of T. evansi and treated 14 days after infection. In the first study 3 groups were injected subcutaneously with Cymelarsan at dose rates of 0.125, 0.25 and 0.50 mg/kg and in the second study 2 i/m at 0.50 and 0.75 mg/kg. The response to treatment was monitored parasitologically by daily microhaematocrit centrifugation technique and weekly mouse inoculation. Enzyme linked immunosorbent assays were used to monitor trypanosomal antibodies and trypanosomal antigens in serum samples collected weekly. Relapse infections occurred in all the groups given the drug s/c whilst all the animals treated i/m remained parasitologically negative up to 80 days after treatment. Results from serological assays, however, suggested the possible persistence of trypanosome infection in the animals treated at a dose rate of 0.50 mg/kg i/m although trypanosomes could not be demonstrated parasitologically. A dose rate of 0.75 mg/kg administered i/m is recommended, therefore, for the treatment of T. evansi infection in Friesian Holstein cattle.

The effect of Trypanosoma evansi infection on oestrous cycling was studied in 12 Friesian Holstein heifers. In the Phase 1 of the investigation, six heifers were infected with T. evansi, the remaining six acted as uninfected controls. Daily body temperature, packed red cell volume (PCV) and parasitaemia measurements were obtained from each animal for 90 days. The animals were examined for external signs of oestrous activity twice daily, blood samples were taken three times a week and subjected to an enzyme-linked immunosorbent assay to detect plasma progesterone. Body weights were measured weekly. Parasites were eliminated by trypanocidal drug treatment 90 days after infection. In Phase 2 of the trial, the uninfected heifers were injected with a different stock of parasites and monitoring was continued as before. Infection with T. evansi resulted in a marked reduction in the rate of weight gain, an increase in body temperatures and a fall in PCV values. Eleven of the heifers continued to cycle normally for the duration of the study, irrespective of their infective status. One animal which stopped cycling lost 16.2% of its pre-infection body weight as a result of the infection and cessation of oestrous activity was considered to have been due to weight loss.

The effect of a single, primary, artificial infection with Trypanosoma evansi was studied in nine Friesian Holstein calves. Body temperature, packed cell volume (PCV) and parasitaemia measurements were obtained daily from each of the infected calves for up to 90 days after parasites were injected, body weights were monitored weekly. T. evansi infection had a marked depressive effect on PCV profiles and the rate of body weight gain.

The effect of controlled amounts of exercise on the outcome of Trypanosoma evansi infection was studied in groups of swamp buffaloes (Bubalus bubalis) experimentally infected with T. evansi. Daily body temperature, packed cell volume (PCV) and parasitaemia measurements were obtained from each animal for up to 110 days after infection. Exercise did not appear to exacerbate the effect of T. evansi infection in that similar temperature, PCV and parasitaemia profiles were obtained with both exercised and rested animals. Trypanosoma evansi infection, however, had a marked effect on temperature and PCV profiles, both of which could adversely affect an infected animal's work output and work tolerance.

Fifteen bovine and 11 buffalo calves born on different farms in a Trypanosoma evansi-endemic area of West Java were monitored for the presence of T. evansi and T. evansi antibody at monthly intervals until they were 12 months of age. Fifty percent of the bovine and 83% of the buffalo calves sampled in the first month of life were antibody positive. This antibody was considered to be of colostral origin. Antibody developing later in life persisted for up to 12 months and was considered to have arisen in response to T. evansi infection. No protective function could be ascribed to the colostral antibody.

Cattle, buffaloes and horses in several areas of Indonesia were examined for evidence of infection with Trypanosoma evansi by the microhaematocrit centrifugation technique (MHCT) and an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to T. evansi. Evidence of infection was found in animals at each sampling site although differences were seen in prevalence rates between sites. Prevalence rates in buffalo were usually higher than in cattle in the same area while in horses they were much lower than in cattle or buffalo. An age-dependent prevalence rate was seen in buffalo and cattle with the highest rates seen in animals older than 2 years. These results concur with the view that T. evansi infection is widespread throughout most of the livestock-producing areas of Indonesia. The apparent lack of any obvious disease owing to T. evansi infection in the sampled animals suggests that a form of stability exists in most endemic areas which serves to ameliorate the effect of T. evansi infection and has an immunological basis linked to the parasite's limited antigenic diversity.

A synthetic tRNA precursor analog containing the structural elements of Escherichia coli tRNA(Phe) was characterized as a substrate for E. coli ribonuclease P and for M1 RNA, the catalytic RNA subunit. Processing of the synthetic precursor exhibited a Mg2+ dependence quite similar to that of natural tRNA precursors such as E. coli tRNA(Tyr) precursor. It was found that Sr2+, Ca2+, and Ba2+ ions promoted processing of the dimeric precursor at Mg2+ concentrations otherwise insufficient to support processing; very similar behavior was noted for E. coli tRNA(Tyr). As noted previously for natural tRNA precursors, the absence of the 3'-terminal CA sequence in the synthetic precursor diminished the facility of processing of this substrate by RNase P and M1 RNA. A study of the Mg2+ dependence of processing of the synthetic tRNA dimeric substrate radiolabeled between C75 and A76 provided unequivocal evidence for an alteration in the actual site of processing by E. coli RNase P as a function of Mg2+ concentration. This property was subsequently demonstrated to obtain (Carter, B. J., Vold, B.S., and Hecht, S. M. (1990) J. Biol. Chem. 265, 7100-7103) for a mutant Bacillus subtilis tRNAHis precursor containing a potential A-C base pair at the end of the acceptor stem.

During an outbreak of trypanosomiasis in cattle, buffalo and horses on the island of Madura, a survey was conducted to determine the prevalence of Trypanosoma evansi in the region. Blood samples were collected from 130 cattle and 147 buffalo in 5 villages, whole blood was examined for trypanosomes by the microhaematocrit (MHCT) method and serum samples were subjected to an enzyme linked immuno absorbent assay to detect T. evansi antibodies. T. evansi was detected by MHCT in 50% of the buffalo examined and 13% of the cattle, while antibodies were detected in 47% of the buffalo and 30% of the cattle. The outbreak was brought under control following the administration of Nagano (Suramin, Bayer) to infected animals and animals considered to be at risk of infection.

A total of 102 susceptible adult Holstein Friesian cattle imported into an area of Paraguay where anaplasmosis and babesiosis are endemic were immunised by infection with Anaplasma centrale and attenuated forms of Babesia bigemina and Babesia bovis obtained from Uruguay. The results indicated that the attenuated forms of both Babesia species protected cattle against heterologous field challenge whereas A. centrale did not invariably confer sufficient protection against a field challenge of A. marginale.

A serological survey was carried out in Paraguay to determine the prevalence of antibodies to Anaplasma marginale, Babesia bigemina and Babesia bovis. The results suggest that the parasites are widely distributed throughout the country with prevalence rates of 92% for A. marginale, 79% for B. bigemina and 71% for B. bovis. Epidemiological studies on calves demonstrated that initial infection with the three blood parasites occurs in most animals within the first nine months of life. It is likely therefore that most of the country can be considered as being enzootically stable with respect to anaplasmosis and babesiosis.

A cell-free protein biosynthesizing system prepared from Escherichia coli CF300 was found to synthesize E. coli tryptophan synthase alpha subunit in a time-dependent manner when programmed with pBN69 plasmid DNA. This plasmid contains the trp promoter from Serratia marcescens adjacent to the coding region of E. coli tryptophan synthase alpha protein [Nichols, B.P., & Yanofsky, C. (1983) Methods Enzymol. 101, 155-164]. The synthesized tryptophan synthase alpha subunit was found to be indistinguishable from authentic alpha subunit protein when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and to have the same specific activity for catalyzing the conversion of indole----L-tryptophan by tryptophan synthase beta 2 subunit, as well as the conversion of indole + glyceraldehyde 3-phosphate to indole-3-glycerol phosphate. In the absence of exogenously added phenylalanine, admixture of E. coli phenylalanyl-tRNAPhe to the protein biosynthesizing system stimulated the production of functional alpha protein; the analogous result was obtained when valine was replaced by E. coli valyl-tRNAVal. The ability of a misacylated tRNA to participate in alpha protein synthesis in this system was established by the use of E. coli phenylalanyl-tRNAVal in the absence of added valine. Protein biosynthesis proceeded normally and gave a product having the approximate molecular weight of tryptophan synthase alpha subunit; as expected, this polypeptide lacked catalytic activity.

Different regions of crown dentin from erupted human premolars were examined ultrastructurally to determine the contents of the dentinal tubules. Odontoblast processes were limited to inner dentin, and nerve fibers were not observed in any tubules examined.

Theileria parva piroplasm infection of mouse erythrocytes was detected, following intraperitoneal inoculation of mice with irradiated cultures of T. parva-infected bovine lymphoid cells. Infection could not be transmitted to cattle using Rhipicephalus appendiculatus ticks.

Uridine 5'-phosphate (UMP) synthase is a multifunctional protein that contains the last two enzyme activities for the de novo biosynthesis of UMP, orotate phosphoribosyltransferase (EC 2.4.2.10) and orotidine-5'-phosphate (OMP) decarboxylase (EC 4.1.1.23). The native enzyme from mouse Ehrlich ascites cells exists in at least three distinct aggregation/conformation states as measured by sedimentation in sucrose gradients: a 3.6S monomer, a 5.1S dimer, and a conformationally altered 5.6S dimer. It has previously been reported that a variety of ligands (of which the most effective is OMP) mediate the conversion of the 3.6S monomer to the two types of dimers. Initial velocity studies with the enzyme in the different native states show that all three forms of UMP synthase have phosphoribosyltransferase activity but that the OMP decarboxylase is either uniquely or at least predominantly associated with the 5.6S form. Activation of this enzyme activity by the substrate appears to be the result of both a dimerization and a conformation step.

Uridine kinase has been purified to homogeneity from Ehrlich ascites tumor cells. For the phosphate acceptor site, the enzyme shows substrate specificity only for ribopyrimidine nucleosides and is active with various analogs that have limited structural alterations; both endocyclic and exocyclic substituents can be acceptable. Of nucleosides that have been used in the chemotherapy of cancer, 5-fluorouridine, 6-azauridine, and 3-deazauridine are good substrates, whereas arabinosylcytosine is a poor substrate. No analogs are better substrates than the physiological substrates uridine and cytidine. 5', 5''' -P1, P4-Bisnucleoside oligophosphate bisubstrate analogs (e.g., Ap4U, Ap5U) were synthesized and tested as inhibitors. The most effective compound was Ap4U; with a Ki of 197 microM, it bound more tightly than ATP but no better than uridine. Ap3A, Ap4A, and Ap5A were also tested, with the result that both Ap4A and Ap4U were most effective, suggesting that this size of bisubstrate analog most closely approaches the spacing of the catalytic site.

Washed erythrocytes infected with Babesia major were mixed with glycerol and snap frozen in liquid nitrogen. After thawing, these cells were used for the preparation of antigen slides for immunofluorescence studies.

Adult Rhipicephalus appendiculatus ticks, infected as nymphs by feeding them on an African buffalo harbouring Theileria lawrencei, were applied to the ears of rabbits. Equal numbers of the feeding ticks were removed daily from rabbits and either dissected and processed so that parasites in their salivary glands could be examined morphologically and classified, or ground in Eagle's Minimal Essential Medium, to harvest parasites from the ticks in the resultant supernatant fluid. The infectivity of the supernatant fluid was tested by inoculation of aliquots into groups of susceptible cattle. It was found that supernatant fluids produced from day 0 to day 2 fed ticks were uninfective to cattle but those produced from 3 to 9 days were infective. The most infective supernatant fluid was produced from 6 day fed ticks. The reactions resulting in cattle were correlated with the number and morphology of parasites in sections of salivary glands and it was found that mature parasites coincided with infectivity of the supernatant fluid to cattle. From the reactions in cattle inoculated with supernatant fluids from 5 and 6 day fed ticks it was concluded that this method of harvesting parasites was efficient. Examination of the salivary glands of a representative sample of a T. lawrencei infected tick batch is a potential means of screening material for T. lawrencei stabilates.

A comparison was made between the development of Theileria mutans (Aitong) in adult Amblyomma variegatum ticks and the development of T. parva (Muguga) in adult Rhipicephalus appendiculatus ticks. Both batches of ticks dropped as engorged nymphs from cattle whose intraerythrocytic parasitemias were approximately 40%, and it was thus possible to make a direct comparison of the infection rates produced in their tick hosts in this experiment by the 2 species of Theileria. Ticks of the 2 batches were fed in ear bags on rabbits, and equal numbers of male and female ticks were removed daily for 5 days. The salivary glands were dissected out from these ticks, and from unfed ones in each batch, processed, and examined for Theileria parasites, which were categorized according to their morphological maturity. The A. variegatum ticks had a lower rate of infection with Theileria. Fewer parasites were observed in the ticks, and they took longer to mature. When the development of the 2 species of Theileria was compared, the major difference appeared to be that T. mutans had large Babesia-like infective particles.

A serological survey was carried out in the Central American state of El Salvador to determine the distribution and prevalence of Anaplasma marginale, Babesia bigemina and Babesia bovis. the results suggest that the parasites are widely distributed throughout the country and prevalence rates of 78.5% for A. marginale, 70.5% for B. bigemina and 73.5% for B. bovis were obtained. Tick counts suggest that boophilus microplus is the main blood parasite vector. Epidemiological studies showed that initial infection with the 3 blood parasites occurs in most animals during the first 7 months of life. Notwithstanding these facts which indicate an area of enzootic stability outbreaks of clinical disease were frequently reported. Twenty-two such outbreaks were investigated and management factors predisposing to outbreaks of clinical disease are discussed.

Uridine kinase from mouse Ehrlich ascites cells can exist in a variety of different aggregation states, from monomer up to aggregates that may contain 32 or more subunits. With very crude enzyme preparations, uridine kinase activity is always associated with several different coexisting molecular weight species. Changes in the aggregation state are produced in the presence of normal effectors (orthophosphate, ATP and CTP) at physiological concentrations. With uridine kinase that has been purified 9,000-fold, enzyme activity is associated with only a single molecular weight species, but is still responsive to the same physiological effectors. In the presence of orthophosphate, uridine kinase has a molecular weight of 380,000 (appropriate for a dodecamer). In the presence of CTP, the enzyme dissociates with concomitant loss of activity. The dissociated enzyme can be reassociated to the native size. These results imply that alteration of the enzyme's quaternary structure by normal effectors constitutes a mechanism for regulating uridine kinase activity in vivo.