An acoustic startle alters knee joint stiffness and neuromuscular control.

Growing evidence suggests that the nervous system contributes to non-contact knee ligament injury, but limited evidence has measured the effect of extrinsic events on joint stability. Following unanticipated events, the startle reflex leads to universal stiffening of the limbs, but no studies have investigated how an acoustic startle influences knee stiffness and muscle activation during a dynamic knee perturbation. Thirty-six individuals were tested for knee stiffness and muscle activation of the quadriceps and hamstrings. Subjects were seated and instructed to resist a 40-degree knee flexion perturbation from a relaxed state. During some trials, an acoustic startle (50 ms, 1000 Hz, 100 dB) was applied 100 ms prior to the perturbation. Knee stiffness, muscle amplitude, and timing were quantified across time, muscle, and startle conditions. The acoustic startle increased short-range (no startle: 0.044 ± 0.011 N·m/deg/kg; average startle: 0.047 ± 0.01 N·m/deg/kg) and total knee stiffness (no startle: 0.036 ± 0.01 N·m/deg/kg; first startle 0.027 ± 0.02 N·m/deg/kg). Additionally, the startle contributed to decreased [vastus medialis (VM): 13.76 ± 33.6%; vastus lateralis (VL): 6.72 ± 37.4%] but earlier (VM: 0.133 ± 0.17 s; VL: 0.124 ± 0.17 s) activation of the quadriceps muscles. The results of this study indicate that the startle response can significantly disrupt knee stiffness regulation required to maintain joint stability. Further studies should explore the role of unanticipated events on unintentional injury.

Do thick leaves avoid thermal damage in critically low wind speeds?

Transient lulls in air movement are rarely measured, but can cause leaf temperature to rise rapidly to critical levels. The high heat capacity of thick leaves can damp this rapid change in temperature. However, little is known about the extent to which increased leaf thickness can reduce thermal damage, or how thick leaves would need to be to have biological significance. We evaluated quantitatively the contribution of small increases in leaf thickness to the reduction in thermal damage during critically low wind speeds under desert conditions. We employed a numerical model to investigate the effect of thickness relative to transpiration, absorptance and leaf size on damage avoidance. We used measured traits and thermotolerance thresholds of real leaves to calculate the leaf temperature response to naturally occurring variable low wind speed. Our results demonstrated that an increase in thickness of only fractions of a millimetre can prevent excursions to damaging high temperatures. This damping effect of increased thickness was greatest when other means of reducing leaf temperature (transpiration, reflectance or reduced size) were lacking. For perennial desert flora, we propose that increased leaf thickness is important in decreasing the incidence of extreme heat stress and, in some species, in enhancing long-term survival.

Ice growth in supercooled solutions of a biological "antifreeze", AFGP 1-5: an explanation in terms of adsorption rate for the concentration dependence of the freezing point.

It is widely accepted, and we agree, that the lowering of the temperature at which ice can grow in a water solution of one of the biological antifreezes is a result of adsorption of the antifreeze molecules at the ice surface. However, how this can produce a well-defined "freezing point" that varies with the solution concentration has remained problematical. The results of a series of measurements of ice growing in supercooled solutions of an effective antifreeze are reported and interpreted in terms of this fundamental problem. It seemed that the solution of the problem would have to rely upon adsorption rate, because that appeared to be the only way for the concentration in solution to be so important. The crystal growth results are most unusual, and appear to confirm this. The growth rates over a wide range of antifreeze concentration in solution (about 0.05 to 9 mg ml(-1)) are zero from the thermodynamic freezing point down to the "non-equilibrium" freezing point, where there is a very sudden increase to a plateau value that then remains about constant as the supercooling is increased by about 2 degrees C. The plateau values of growth rate are faster than those from pure water at the lower-supercooling ends of the plateaus, but slower at higher supercooling, until the growth rate starts rising toward that from pure water. These plateau values of growth rate increase markedly with increasing concentration of the antifreeze in solution. Along with these changes there are complex changes in the growth orientations, from c-axis spicules in the plateaus to those more characteristic of growth from pure water at greater supercooling. We conclude that the non-equilibrium freezing point is determined by the adsorption rate. It is the warmest temperature at which the ice growth rate on the basal plane (where the antifreeze does not adsorb) is fast enough to prevent the area of basal face on a growing ice crystal from becoming too small to grow, which is determined in turn by the adsorption rate on non-basal surfaces, which is proportional to the solution concentration. This mechanism answers the question of how the antifreeze stops growth rather than how it prevents growth, a subtle but important difference.

Superficial motor units are larger than deeper motor units in human vastus lateralis muscle.

Previous studies have suggested that regionalization may occur for human motor units, whereby smaller motor units are located in deeper parts of the muscle and larger motor units are located in more superficial portions. We examined this possibility in the human vastus lateralis muscle using macro-EMG (electromyography) to estimate motor unit size. The sample consisted of nine individuals from whom 114 motor units were recorded at forces ranging between 5% and 60% MVC. Peak-to-peak macro-EMG amplitude was well correlated with macro area (Spearman rho = 0.96). There was a statistically significant inverse relationship between recording depth and macro peak-to-peak amplitude (rho = -0.402, p < 0.001). We conclude that there is a nonrandom distribution of motor units in human muscle, with larger motor units located in more superficial regions and smaller units located in deeper regions. Clinicians who monitor motor unit activity need to recognize that a representative sample of motor unit recordings should include motor units from both deeper and more superficial regions of muscle.

Beverage caffeine intake in US consumers and subpopulations of interest: estimates from the Share of Intake Panel survey.

Concerns exist about the potential adverse health effects of high consumption of dietary caffeine, especially in children and pregnant women. Recommended caffeine intakes corresponding to no adverse health effects have been suggested recently for healthy adults (400-450 mg/day), for women contemplating pregnancy (300 mg/day), and for young children age 4-6 years (45 mg/day). To determine whether current caffeine intake approaches these levels, intake from major dietary sources (coffee, tea and carbonated soft drinks) were measured in 10,712 caffeinated beverage consumers in the 1999 US Share of Intake Panel, a targeted beverage survey. Mean caffeine intakes in adult caffeinated beverage consumers ranged from 106 to 170 mg/day (90th percentile intake was 227-382 mg/day). In children 1-5 and 6-9 years, mean caffeine intakes were 14 and 22 mg/day, respectively; corresponding 90th percentile intakes were 37 and 45 mg/day. Pregnant women consumed an average of 58 mg/day (157 mg/day at the 90th percentile), and women of reproductive age ingested 91-109 mg/day (229-247 mg/day at the 90th percentile). These data show that while mean caffeine intakes are within recommended safe levels, heavy consumers of certain subpopulations, including young children and women contemplating pregnancy, might benefit from dietary advice.

Adaptations in muscular activation of the knee extensor muscles with strength training in young and older adults.

The purpose of this study was to compare the extent of muscular activation during maximal voluntary knee extension contractions in old and young individuals and to examine the effects of resistance training on muscular activation in each group. The interpolated twitch technique was used to estimate muscular activation during two pre-training baseline tests, and after two and six weeks of resistance training. Throughout the study, the older group was 30% less strong than the young group (p=0.02). The training protocol was effective in both groups with overall isometric strength gains of 30 and 36% in the older (p=0.01) and young (p<0.01) groups, respectively. 10-RM training loads increased by 66% in the old group (p<0.01) and by 77% in the young group (p<0.01) throughout training. At the first baseline test, a 2% difference in muscular activation between groups (p=0.3) did not explain the large disparity in strength. Muscular activation increased by 2% in both groups throughout training (p<0.01). Despite considerably less muscular strength in the older group, muscular activation was greater than 95% of maximum and appears to be equal in both young and older individuals. Both groups demonstrated similar but small increases in muscular activation throughout training.

Effect of superficial heat, deep heat, and active exercise warm-up on the extensibility of the plantar flexors.

BACKGROUND AND PURPOSE: Warm-up prior to static stretching enhances muscle extensibility. The relative effectiveness of different modes of warm-up, however, is unknown. The purpose of this study was to evaluate the effectiveness of superficial heat, deep heat, and active exercise warm-up prior to stretching compared with stretching alone on the extensibility of the plantar-flexor muscles. SUBJECTS: Ninety-seven subjects (59 women, 38 men) with limited dorsiflexion range of motion (ROM) were randomly assigned to 1 of 5 groups. Female subjects had a mean age of 27.6 years (SD=7.68, range=17-50), and male subjects had a mean age of 26.8 years (SD=6.87, range=18-48). METHODS: The first group (group 1) was a control group and did not perform the stretching protocol. The 4 experimental groups (groups 2-5) performed a stretching protocol 3 days per week for 6 weeks. Group 2 performed the static stretching protocol only; group 3 performed active heel raises before stretching; group 4 received 15 minutes of superficial, moist heat to the plantar-flexor muscles before stretching; and group 5 received continuous ultrasound for 7 minutes before stretching. Dorsiflexion ROM measurements were taken initially and after 2, 4, and 6 weeks. RESULTS: All experimental groups increased active and passive range of motion (AROM and PROM). The mean AROM/PROM differences at 6 weeks were 1.11/1.39 degrees for group 1, 4.10/6.11 degrees for group 2, 4.16/4.21 degrees for group 3, 4.38/4.90 degrees for group 4, and 6.20/7.35 degrees for group 5. The group receiving ultrasound before performing the stretching protocol (group 5) displayed the greatest increase in both AROM (6.20() and PROM (7.35(). DISCUSSION AND CONCLUSION: Among the modalities tested, the use of ultrasound for 7 minutes prior to stretching may be the most effective for increasing ankle dorsiflexion ROM.

Effect of ketoprofen on muscle function and sEMG activity after eccentric exercise.

PURPOSE: This study examined whether ketoprofen, a nonsteroidal anti-inflammatory drug, attenuated muscle soreness (SOR), improved maximal isometric force (MIF) recovery, and/or altered myoelectric activity after high-force eccentric exercise. METHODS: 48 subjects were randomly assigned to one of four groups: CON: no exercise/no drug (N = 12); PLA: exercise + placebo (N = 12); TRT-100: exercise + 100 mg oral ketoprofen (N = 12); and TRT-25: exercise + 25 mg oral ketoprofen (N = 12). PLA, TRT-100, and TRT-25 were administered in a double-blind fashion. Baseline measurements of SOR, MIF, and surface electromyographic (EMG) amplitude were taken, and PLA, TRT-100, and TRT-25 performed 50 maximal eccentric contractions of the elbow flexors; 36 h later, subjects reporting moderate soreness were given ketoprofen or placebo and SOR measures were taken hourly for 8 h. EMG amplitude was assessed during MIF before dosing and again 8 h later and during submaximal contractions of 5%, 10%, and 20% of MIF before dosing and hourly for 8 h. RESULTS: Eccentric exercise increased myoelectric activity during submaximal force measurements in PLA, TRT-100, and TRT-25 in all conditions. Ketoprofen had no effect on reducing this increase in EMG activity. Ketoprofen attenuated perceived SOR (P < 0.05) and enhanced MIF recovery (P < 0.05) compared with placebo. TRT-100 and TRT-25 demonstrated 10% and 19% reductions in SOR, respectively, and 16% and 9% increases in MIF, respectively, whereas PLA demonstrated a 1% increase in SOR and 9% decrease in MIF over 8 h. CONCLUSION: Ketoprofen treatment after muscle damaging exercise reduces muscle soreness and improves force recovery.

Correlated evolution of chloroplast heat shock protein expression in closely related plant species.

Interspecific variation in chloroplast low molecular weight (cLMW) HSP (heat shock protein) expression was examined with respect to phylogeny, species specific leaf area, chlorophyll fluorescence, and mean environmental conditions within species ranges. Eight species of Ceanothus (Rhamnaceae) were heat shocked for 4 h at several different temperatures. Leaf samples were collected immediately after the heat shock, and cLMW HSP expression was quantified using Western blots. At 45°C species from the subgenus Cerastes had significantly greater cLMW HSP expression than species from the subgenus Ceanothus. Specific leaf area was negatively correlated with cLMW HSP expression after the 45°C heat treatment. In addition, chlorophyll fluorescence (F(v)/F(m)) 1 h after the heat shocks was positively correlated with cLMW HSP expression. Contrary to our prediction, there was no correlation between July maximum temperature within species ranges and cLMW HSP expression. These results suggest that evolutionary differentiation in cLMW HSP expression is associated with leaf physiological parameters and related aspects of life history, yet associations between climatic conditions within species ranges and cLMW HSP expression require further study.

Muscular and metabolic costs of uphill backpacking: are hiking poles beneficial?

PURPOSE: The purpose of the present study was to compare pole and no-pole conditions during uphill backpacking, which was simulated on an inclined treadmill with a moderately heavy (22.4 kg, 30% body mass) backpack. METHODS: Physiological measurements of oxygen consumption, heart rate, and RPE were taken during 1 h of backpacking in each condition, along with joint kinematic and electromyographic comparisons from data collected during a third test session. RESULTS: The results showed that although imposing no metabolic consequence, pole use elicited a longer stride length (1.27 vs 1.19 m), kinematics that were more similar to those of unloaded walking, and reduced activity in several lower extremity muscles. Although pole use evoked a greater heart rate (113.5 vs 107 bpm), subjects were backpacking more comfortably as indicated by their ratings of perceived exertion (10.8 vs 11.6). The increased cardiovascular demand was likely to support the greater muscular activity in the upper extremity, as was observed in triceps brachii. CONCLUSION: By redistributing some of the backpack effort, pole use alleviated some stress from the lower extremities and allowed a partial reversal of typical load-bearing strategies.

Vacuum-assisted closure versus saline-moistened gauze in the healing of postoperative diabetic foot wounds.

Diabetic foot wounds present a great challenge to wound care practitioners. The objective of this pilot study was to determine whether vacuum-assisted closure (V.A.C.) therapy would afford quicker wound resolution as compared to saline-moistened gauze in the treatment of postoperative diabetic foot wounds. Ten patients were randomized into either the experimental V.A.C. group or control saline gauze group. Included in the study were diabetic patients 18 to 75 years of age who had a nonhealing foot ulceration. Excluded were those patients with venous disease, coagulopathy, or those who had active infections not resolved by initial surgical debridement. All foot ulcers were surgically debrided prior to initiation of V.A.C. or gauze treatment. In the experimental group, V.A.C. dressings were applied in accordance with manufacturer's protocol for chronic wounds and changed every 48 hours. In the control group, saline gauze dressings were applied at the time of surgical debridement and changed twice a day thereafter. Measurements and photos were obtained to document wound progress. Main outcome measures included: 1) time to satisfactory healing (calculated from date of initial debridement to date of definitive closure, and 2) change in wound surface area (calculated from initial wound tracing to final tracing). Satisfactory healing in the V.A.C. group was achieved in 22.8 (+/- 17.4) days, compared to 42.8 (+/- 32.5) days in the control group. Surface area changes of 28.4% (+/- 24.3) average decrease in wound size in the V.A.C. group, compared to a 9.5% (+/- 16.9) average increase in the control group during measurement period.

Structure-function relationship in the antifreeze activity of synthetic alanine-lysine antifreeze polypeptides.

Recently antifreeze proteins (AFP) have been the subject of many structure-function relationship studies regarding their antifreeze activity. Attempts have been made to elucidate the structure-function relationship by various amino acid substitutions, but to our knowledge there has been no successful from first principles design of a polypeptide that would bind to designated ice planes along a specific direction. In this paper we show the results of our first attempt on an entirely de novo design of an alanine-lysine-rich antifreeze polypeptide. This 43 residue alanine-lysine peptide exhibits characteristic nonequilibrium freezing point depression and binds to the designated (210) planes of ice along the [122] vector. The structural and thermodynamic properties of this polypeptide were determined using circular dichroism spectroscopy and its nonequilibrium antifreeze properties were investigated using an ice-etching method and nanoliter osmometry.

Valine substituted winter flounder 'antifreeze': preservation of ice growth hysteresis.

Three mutant polypeptides of the type I 37-residue winter flounder 'antifreeze' protein have been synthesized. All four threonine residues in the native peptide were been mutated to serine, valine and glycine respectively and two additional salt bridges were incorporated into the sequences in order to improve aqueous solubility. The peptides were analyzed by nanoliter osmometry, the 'ice hemisphere' test, the 'crystal habit' test, measurement of ice growth hysteresis and CD spectroscopy. While the valine and serine mutants retain the alpha-helical structure, only the valine mutant retains 'antifreeze' activity similar to that of the native protein. These data show that the threonine hydroxyl groups do not play a crucial role in the accumulation of the native 'antifreeze' protein at the ice/water interface and the inhibition of ice growth below the equilibrium melting temperature.

Analysis of shorthorn sculpin antifreeze protein stereospecific binding to (2-1 0) faces of ice.

In this paper we report the results of our studies on the stereospecific binding of shorthorn sculpin antifreeze protein (AFP) to (2 -1 0) secondary prism faces of ice. Using ice crystal growth and etching techniques together with molecular modeling, molecular dynamics, and energy minimization, we explain the nature of preferential binding of shorthorn sculpin AFP along the [1 2 2] direction on (2- 1 0) planes. In agreement with ice etching studies, the mechanism of preferential binding suggested by molecular modeling explains why the binding of shorthorn sculpin AFP occurs along [1 2 2] and not along its mirror symmetry-related direction [-1 -2 2] on (2 -1 0). This binding mechanism is based on the protein-crystal surface enantioselective recognition that utilizes both alpha-helical protein backbone matching to the (2 -1 0) surface topography and matching of side chains of polar/charged residues with specific water molecule positions in the ice surface. The mechanisms of winter flounder and shorthorn sculpin antifreeze binding to ice are compared.

Nonequilibrium antifreeze peptides and the recrystallization of ice.

Evidence is presented that the nonequilibrium antifreeze peptide (AFP) from winter flounder has a special ability to inhibit recrystallization in ice only when an appreciable amount of liquid is present, as is the case when the system contains salts and the temperature is not too low. In this circumstance the AFP binds to the ice surface at the ice-solution interfaces in grain boundaries, preventing migration of the solution and effectively immobilizing the boundaries. In the absence of liquid, recrystallization inhibition appears to be a common property of many peptides. This is consistent with the view that the special effects of AFPs require a structural fit onto ice, and therefore require the AFP molecules to have the mobility to achieve that fit. Since the concentration of salt required to induce the special recrystallization inhibition effects of AFPs is lower (< 10 mM) than that found normally in physiological fluids, AFPs could play a role in the survival of organisms by preventing damage due to recrystallization. The proposition that mobility is needed for AFP molecules to produce their special influence upon ice growth argues against any special effects of AFPs in devitrification.

Adsorption to ice of fish antifreeze glycopeptides 7 and 8.

Experimental results show that fish antifreeze glycopeptides (AFGPs) 8 and 7 (with 4 and 5 repeats respectively of the Ala-Ala-Thr backbone sequence) bond onto ice prism planes aligned along a-axes, and inhibit crystal growth on prism planes and on surfaces close to that orientation. The 9.31-A repeat spacing of the AFGP in the polyproline II helix configuration, deduced from NMR studies, matches twice the repeat spacing of ice in the deduced alignment direction, 9.038 A, within 3%. A specific binding model is proposed for the AFGP and for the alpha-helical antifreeze peptide of winter flounder. For AFGP 7-8, two hydroxyl groups of each disaccharide (one disaccharide is attached to each threonine) reside within the ice surface, so that they are shared between the ice crystal and the disaccharide. This provides 24 hydrogen bonds between AFGP 8 and the ice and 30 for AFGP 7, explaining why the chemical adsorption is virtually irreversible and the crystal growth can be stopped virtually completely. The same scheme of sharing polar groups with the ice works well with the alpha-helical antifreeze of winter flounder, for which an amide as well as several hydroxyls are shared. The sharing of polar groups with the ice crystal, rather than hydrogen-bonding to the ice surface, may be a general requirement for adsoprtion-inhibition of freezing.

Plant thermal hysteresis proteins.

Proteins which produce a thermal hysteresis (i.e. lower the freezing point of water below the melting point) are common antifreezes in cold adapted poikilothermic animals, especially fishes from ice-laden seas and terrestrial arthropods. However, these proteins have not been previously identified in plants. 16 species of plants collected from northern Indiana in autumn and winter had low levels of thermal hysteresis activity, but activity was absent in summer. This suggests that thermal hysteresis proteins may be a fairly common winter adaptation in angiosperms. Winter stem fluid from the bittersweet nightshade, Solanum dulcamara L., also showed the recrystallization inhibition activity characteristic of the animal thermal hysteresis proteins (THPs), suggesting a possible function for the THPs in this freeze tolerant species. Other potential functions are discussed. Antibodies to an insect THP cross reacted on immunoelectroblots with proteins in S. dulcamara stem fluid, indicating common epitopes in the insect and plant THPs.

Small cell carcinoma of the ovary.

A case of a 29-year-old woman with primary small cell carcinoma of the ovary associated with hypercalcemia is presented. Despite intensive surgical and chemotherapeutic intervention, the patient expired after 19 months. Literature review indicates that this rare tumor grows rapidly, with hypercalcemia occurring frequently. Absence of effective treatment results in early mortality.

Adsorption of alpha-helical antifreeze peptides on specific ice crystal surface planes.

The noncolligative peptide and glycopeptide antifreezes found in some cold-water fish act by binding to the ice surface and preventing crystal growth, not by altering the equilibrium freezing point of the water. A simple crystal growth and etching technique allows determination of the crystallographic planes where the binding occurs. In the case of elongated molecules, such as the alpha-helical peptides in this report, it also allows a deduction of the molecular alignment on the ice surface. The structurally similar antifreeze peptides from winter flounder (Pseudopleuronectes americanus) and Alaskan plaice (Pleuronectes quadritaberulatus) adsorb onto the (2021) pyramidal planes of ice, whereas the sculpin (Myoxocephalus scorpius) peptide adsorbs on (2110), the secondary prism planes. All three are probably aligned along (0112). These antifreeze peptides have 11-amino acid sequence repeats ending with a polar residue, and each repeat constitutes a distance of 16.5 A along the helix, which nearly matches the 16.7 A repeat spacing along (0112) in ice. This structural match is undoubtedly important, but the mechanism of binding is not yet clear. The suggested mechanism of growth inhibition operates through the influence of local surface curvature upon melting point and results in complete inhibition of the crystal growth even though individual antifreeze molecules bind at only one interface orientation.