Colony-stimulating factor-1 (CSF-1) directly inhibits receptor activator of nuclear factor-{kappa}B ligand (RANKL) expression by osteoblasts.

Colony-stimulating factor-1 (CSF-1), released by osteoblasts, stimulates the proliferation of osteoclast progenitors via the c-fms receptor (CSF-1R) and, in combination with receptor activator of nuclear factor-kappaB ligand (RANKL), leads to the formation of mature osteoclasts. Whether the CSF-1R is expressed by osteoblasts and mediates specific biological effects in osteoblasts has not been explored. Wild-type primary calvaria osteoblasts (OB) were analyzed for CSF-1R expression (RT-PCR and Western blot) and functionality (immunocomplex kinase assay). OB were serum starved for 24 h, and the effect of CSF-1 (0-100 ng/ml) on OB biological activities was determined at 48 h. In wild-type mouse bone marrow cultures, CSF-1 was tested for its effect on RANKL mRNA and osteoclast formation. Because ROS influence osteoblast RANKL expression, studies analyzed the effect of CSF-1 on reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity and Nox1 and Nox4 proteins. Results indicate that OB express CSF-1R mRNA and protein and that CSF-1R could be phosphorylated in the presence of CSF-1. In osteoblasts, CSF-1 decreased RANKL mRNA in a dose- and time-dependent manner. Incubation of bone marrow cultures with CSF-1 resulted in a significant decline in tartrate-resistant acid phosphatase (TRACP) activity and CTR expression. RANKL-decreased expression by CSF-1 was correlated with a decrease of NADPH oxidase activity as well as Nox1 and Nox4 protein levels. These findings provide the first evidence that osteoblasts express CSF-1R and are a target for CSF-1 ligand. CSF-1-mediated inhibition of RANKL expression on osteoblasts may provide an important mechanism for coupling bone formation/resorption and preventing excessive osteoclastogenesis during normal skeletal growth.

Feto-placental adaptations to maternal obesity in the baboon.

Maternal obesity is present in 20-34% of pregnant women and has been associated with both intrauterine growth restriction and large-for-gestational age fetuses. While fetal and placental functions have been extensively studied in the baboon, no data are available on the effect of maternal obesity on placental structure and function in this species. We hypothesize that maternal obesity in the baboon is associated with a maternal inflammatory state and induces structural and functional changes in the placenta. The major findings of this study were: 1) decreased placental syncytiotrophoblast amplification factor, intact syncytiotrophoblast endoplasmic reticulum structure and decreased system A placental amino acid transport in obese animals; 2) fetal serum amino acid composition and mononuclear cells (PBMC) transcriptome were different in fetuses from obese compared with non-obese animals; and 3) maternal obesity in humans and baboons is similar in regard to increased placental and adipose tissue macrophage infiltration, increased CD14 expression in maternal PBMC and maternal hyperleptinemia. In summary, these data demonstrate that in obese baboons in the absence of increased fetal weight, placental and fetal phenotype are consistent with those described for large-for-gestational age human fetuses.

High d(+)glucose concentration inhibits RANKL-induced osteoclastogenesis.

Diabetes is a chronic disease associated with hyperglycemia and altered bone metabolism that may lead to complications including osteopenia, increased risk of fracture and osteoporosis. Hyperglycemia has been implicated in the pathogenesis of diabetic bone disease; however, the biologic effect of glucose on osteoclastogenesis is unclear. In the present study, we examined the effect of high d(+)glucose (d-Glc) and l(-)glucose (l-Glc; osmotic control) on RANKL-induced osteoclastogenesis using RAW264.7 cells and Bone Marrow Macrophages (BMM) as models. Cells were exposed to sustained high glucose levels to mimic diabetic conditions. Osteoclast formation was analyzed using tartrate resistant acid phosphatase (TRACP) assay, expression of calcitonin receptor (CTR) and cathepsin K mRNAs, and cultures were examined for reactive oxygen species (ROS) using dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, caspase-3 and Nuclear Factor kappaB (NF-kappaB) activity. Cellular function was assessed using a migration assay. Results show, for the first time, that high d-Glc inhibits osteoclast formation, ROS production, caspase-3 activity and migration in response to RANKL through a metabolic pathway. Our findings also suggest that high d-Glc may alter RANKL-induced osteoclast formation by inhibiting redox-sensitive NF-kappaB activity through an anti-oxidative mechanism. This study increases our understanding of the role of glucose in diabetes-associated bone disease. Our data suggest that high glucose levels may alter bone turnover by decreasing osteoclast differentiation and function in diabetes and provide new insight into the biologic effects of glucose on osteoclastogenesis.

A method to study lumbar spine response to axial compression during magnetic resonance imaging: technical note.

STUDY DESIGN: A magnetic resonance imaging (MRI)-compatible device was developed to apply calibrated compression loads to the lumbar spine during imaging. Experiments were performed to establish a protocol to measure lumbar load-response and estimate muscle-force contribution to spinal load. OBJECTIVE: To develop experimental methodology for direct study of lumbar spine response to compression load. SUMMARY OF BACKGROUND DATA: Most lumbar MRI scans require subjects to lie relaxed and supine, but spinal stenosis has been demonstrated to increase during moderate compressive loading. Several devices have been used to load the spine during MRI, but they could not maintain and/or change calibrated loads during MRI experiments. Furthermore, artifact from viscoelastic creep during imaging was not considered. METHODS: An MRI-compatible spine compression unit with pneumatic load elements was developed to produce calibrated compression loads. Young healthy men were loaded with 140% body weight for up to 10 minutes to establish an appropriate test protocol. Muscle force contribution to spinal load was estimated from electromyography experiments. RESULTS: The spine compression unit produced specified loads +/- 29 N (standard deviation). Spine viscoelastic creep reached steady state by 6.5 minutes, leaving 3.5 minutes for image acquisition. The subjects could support 1.0 body weight for the requisite 10 minutes. Muscle compressive force estimates were only 135 N during application of 1.4 x body weight external compression load; thus, internal muscle forces during supine spine compression could be neglected. CONCLUSIONS: The lumbar load/image protocol fits within the time constraints of creep deformation and subject endurance. These methods allow acute lumbar mechanical response measurements during loading.

Relationships between throwing mechanics and shoulder distraction in professional baseball pitchers.

The extreme forces and torques and the high speeds and excessive ranges of motion of baseball pitching place tremendous stress on the soft tissues of the throwing shoulder. Little is known about the relationship between pitching mechanics and shoulder joint stress, especially in professional athletes. The purpose of this study was to quantify joint loads and kinematic parameters of pitching mechanics at the major league level and to study their relationships. Three-dimensional, high-speed video data were collected on 40 professional pitchers during the 1998 Cactus League spring training. A clinically significant distraction force was calculated at the shoulder joint, which reached an average peak value of 947 +/- 162 N (108% +/- 16% body weight). Descriptive statistics and a multiple linear regression analysis were used to relate shoulder distraction to kinematic and kinetic parameters of pitching mechanics. This study was undertaken not only to investigate the peak forces and torques on the shoulder, but also to identify potential areas of intervention that might prevent throwing injuries. Knowledge of joint ranges of motion, angular velocities, and joint-reaction forces can provide a scientific basis for improved preventive and rehabilitative protocols for baseball pitchers.

The effects of extended play on professional baseball pitchers.

The purpose of this study was to investigate kinematic and kinetic changes as a result of extended play in baseball pitching. Seven major league baseball pitchers were videotaped with high-speed (120 Hz) cameras during multiple innings of the same game. For each athlete, two fastballs (one thrown during the initial inning of play and one from the final inning) were chosen for analysis. Twenty-one physical landmarks were manually digitized from the video data. Kinematic and kinetic parameters were subsequently calculated relative to four phases of the pitching motion: windup, cocking, acceleration, and follow-through. Paired t-tests revealed that seven parameters changed significantly between early and late innings. These included decreases in maximum external rotation of the shoulder, knee angle at ball release, ball velocity, maximum distraction force at both the shoulder and elbow, and horizontal adduction torque at both release and its maximum value. Ultimately, a decline in performance was evident by a 2 m/s (5 mph) drop in ball speed. It is unclear whether the kinematic and kinetic changes occurred because of fatigue or if protective mechanisms were adopted.

The mild cleavage of 2-amino-2-deoxy-D-glucoside methoxycarbonyl derivatives.

The conversion of methyl carbamate to the corresponding free amine is described for a series of 2-amino-2-deoxy-D-glucosamine derivatives. Cleavage of methoxycarbonyl moiety with MeSiCl(3) and triethylamine in dry THF at 60 degrees C and subsequent aqueous hydrolysis yields the free amine in 54 to 93% yields. The selective cleavage of methyl carbamates with MeSiCl(3) in the presence of a 2,2,2-trichloroethoxycarbonyl group or 2-azido glycosides affords selectively, orthogonal N-deprotected carbohydrates.

Routine determination of sulfonylurea, imidazolinone, and sulfonamide herbicides at nanogram-per-liter concentrations by solid-phase extraction and liquid chromatography/mass spectrometry.

Sulfonylurea (SU), imidazolinone (IMI), and sulfonamide (SA) herbicides are new classes of low-application-rate herbicides increasingly used by farmers. Some of these herbicides affect both weed and crop species at low dosages and must be carefully used. Less is known about the effect of these compounds on non-crop plant species, but a concentration of 100 ng/l in water has been proposed as the threshold for possible plant toxicity for most of these herbicides. Hence, analytical methods must be capable of detecting SUs, IMIs, and SAs at concentrations less than 100 ng/l in ambient water samples. The authors developed a two-cartridge, solid-phase extraction method for isolating 12 SU, 3 IMI, and 1 SA herbicides by using high-performance liquid chromatography/electrospray ionization-mass spectrometry (HPLC/ESI-MS) to identify and quantify these herbicides to 10 ng/l. This method was used to analyze 196 surface- and ground-water samples collected from May to August 1998 throughout the Midwestern United States, and more than 100 quality-assurance and quality-control samples. During the 16 weeks of the study, the HPLC/ESI-MS maintained excellent calibration linearity across the calibration range from 5 to 500 ng/l, with correlation coefficients of 0.9975 or greater. Continuing calibration verification standards at 100-ng/l concentration were analyzed throughout the study, and the average measured concentrations for individual herbicides ranged from 93 to 100 ng/l. Recovery of herbicides from 27 reagent-water samples spiked at 50 and 100 ng/l ranged from 39 to 92%, and averaged 73%. The standard deviation of recoveries ranged from 14 to 26%, and averaged 20%. This variability reflects multiple instruments, operators, and the use of automated and manual sample preparation. Spiked environmental water samples had similar recoveries, although for some herbicides, the sample matrix enhanced recoveries by as much as 200% greater than the spiked concentration. This matrix enhancement was sample- and compound-dependent. Concentrations of herbicides in unspiked duplicate environmental samples were typically within 25% of each other. The results demonstrate the usefulness of HPLC/ESI-MS for determining low-application-rate herbicides at ambient concentrations.

Determination of submicrogram-per-liter concentrations of caffeine in surface water and groundwater samples by solid-phase extraction and liquid chromatography.

A method for determining submicrogram-per-liter concentrations of caffeine in surface water and groundwater samples has been developed. Caffeine is extracted from a 1 L water sample with a 0.5 g graphitized carbon-based solid-phase cartridge, eluted with methylene chloride-methanol (80 + 20, v/v), and analyzed by liquid chromatography with photodiode-array detection. The single-operator method detection limit for organic-free water samples was 0.02 microgram/L. Mean recoveries and relative standard deviations were 93 +/- 13% for organic-free water samples fortified at 0.04 microgram/L and 84 +/- 4% for laboratory reagent spikes fortified at 0.5 microgram/L. Environmental concentrations of caffeine ranged from 0.003 to 1.44 micrograms/L in surface water samples and from 0.01 to 0.08 microgram/L in groundwater samples.

Biomechanics of wrist injuries in sports.

Athletic injuries of the wrist are common. Wrist anatomy, biomechanics, and injury mechanisms are outlined for the athletic population. Common injuries are discussed in a biomechanical context for impact sports; racquet, stick, and club sports; and apparatus and external contact sports.

Biomechanics of the elbow during baseball pitching.

By understanding pitching biomechanics, therapists can develop better preventive and rehabilitative programs for pitchers. The purpose of this study was to quantify and explain the joint motions, loads, and muscle activity that occur at the elbow during baseball pitching. Seven healthy, adult pitchers were examined with synchronized high-speed video digitization and surface electromyography. Elbow extension before ball release corresponded with a decrease in biceps activity and an increase in triceps activity. A varus torque of 120 Nm, acting to resist valgus stress, occurred near the time of maximum shoulder external rotation. Previous cadaveric research showed that the ulnar collateral ligament by itself cannot withstand a valgus load of this magnitude. Triceps, wrist flexorpronator, and anconeus activity during peak valgus stress suggests that these muscles may act as dynamic stabilizers to assist the ulnar collateral ligament in preventing valgus extension overload.