Torque complexity of maximal knee extensor isometric contraction in individuals following anterior cruciate ligament reconstruction.

BACKGROUND: Current rehabilitation goals following anterior cruciate ligament reconstruction are structured around the maximal force generating capabilities of the muscle. Force fluctuations, an index of force control, have been observed to alter post- anterior cruciate ligament reconstruction. The temporal structure, or "complexity" of force fluctuations may provide important insight into the post-operative muscular recovery. The aims of this study were 1) to compare quadriceps torque complexity in anterior cruciate ligament reconstructed patients to the contralateral limb and to healthy, controls and 2) to assess the relationships between torque complexity to patient outcomes. METHODS: Data from 120 anterior cruciate ligament reconstructed participants (65 Females, 21.0 ± 8.3 years, 5.96 ± 0.48-months post-surgery) and 95 healthy controls (50 Females, 21.5 ± 2.9 years) were collected. A 30-s knee extensor maximal isometric contraction was completed to calculate approximate entropy, a measure of torque complexity. FINDINGS: Approximate entropy was found to decrease throughout the 30-s trial (P < .001, Cohen's d = 1.87 [1.64,2.10]). The anterior cruciate ligament reconstructed limb demonstrated greater approximate entropy compared to the contralateral limb or to healthy controls (P < .001, Cohen's d = 0.64 [0.38,0.90]). approximate entropy at the end of the trial demonstrated weak, negatively relationships with peak torque, patient reported outcome measures, and knee extensor fatigue (r = -0.21 to -0.32, P < .05). INTERPRETATION: A greater torque complexity in individuals following anterior cruciate ligament reconstruction was weakly related to lower quadriceps strength, lower subjective function, and quadriceps fatigue resistance. The complexity of force fluctuations during a sustained maximal task may draw clinical insight into the recovery of motor function following anterior cruciate ligament reconstruction.

Agriculture causes nitrate fertilization of remote alpine lakes.

Humans have altered Earth's nitrogen cycle so dramatically that reactive nitrogen (Nr) has doubled. This has increased Nr in aquatic ecosystems, which can lead to reduced water quality and ecosystem health. Apportioning sources of Nr to specific ecosystems, however, continues to be challenging, despite this knowledge being critical for mitigation and protection of water resources. Here we use Δ(17)O, δ(18)O and δ(15)N from Uinta Mountain (Utah, USA) snow, inflow and lake nitrate in combination with a Bayesian-based stable isotope mixing model, to show that at least 70% of nitrates in aquatic systems are anthropogenic and arrive via the atmosphere. Moreover, agricultural activities, specifically nitrate- and ammonium-based fertilizer use, are contributing most (∼60%) Nr, and data from other North American alpine lakes suggest this is a widespread phenomenon. Our findings offer a pathway towards more effective mitigation, but point to challenges in balancing food production with protection of important water resources.

HbA 1c as a risk factor for heart failure in persons with diabetes: the Atherosclerosis Risk in Communities (ARIC) study.

AIMS/HYPOTHESIS: Heart failure (HF) incidence in diabetes in both the presence and absence of CHD is rising. Prospective population-based studies can help describe the relationship between HbA(1c), a measure of glycaemia control, and HF risk. METHODS: We studied the incidence of HF hospitalisation or death among 1,827 participants in the Atherosclerosis Risk in Communities (ARIC) study with diabetes and no evidence of HF at baseline. Cox proportional hazard models included age, sex, race, education, health insurance status, alcohol consumption, BMI and WHR, and major CHD risk factors (BP level and medications, LDL- and HDL-cholesterol levels, and smoking). RESULTS: In this population of persons with diabetes, crude HF incidence rates per 1,000 person-years were lower in the absence of CHD (incidence rate 15.5 for CHD-negative vs 56.4 for CHD-positive, p<0.001). The adjusted HR of HF for each 1% higher HbA(1c) was 1.17 (95% CI 1.11-1.25) for the non-CHD group and 1.20 (95% CI 1.04-1.40) for the CHD group. When the analysis was limited to HF cases which occurred in the absence of prevalent or incident CHD (during follow-up) the adjusted HR remained 1.20 (95% CI 1.11-1.29). CONCLUSIONS/INTERPRETATIONS: These data suggest HbA(1c) is an independent risk factor for incident HF in persons with diabetes with and without CHD. Long-term clinical trials of tight glycaemic control should quantify the impact of different treatment regimens on HF risk reduction.

Determinants of gait as applied to children with cerebral palsy.

In the present study, we quantified the isolated contributions of eight determinants of gait on the vertical center of mass (CoM) displacement of both typically developing children and children with cerebral palsy (CP). The role of these determinants, on vertical excursion, has never been examined for children or children with CP. We hypothesized that the relative contributions of the determinants to vertical CoM excursion of children with CP would be the same as the age-matched controls. We found that based on the similarities in the determinants effect on gait between the controls and adults reflect that children of this age walk with a mature gait. When applied to subjects with CP the determinant analysis found similar, but slightly exaggerated effects of those of the controls. All determinants that negatively affect CoM excursion were significantly worse in the children with CP, while those determinants that decreased excursion were varied. Heel rise, single support knee flexion, and pelvic obliquity had similar effects for on both groups. Pelvic rotation resulted in more excursion reduction in the controls, while leg inclination was more beneficial in reducing the CP groups excursion. The main cause for increased vertical excursion of the CoM in the children with CP was the increased knee flexion of both legs during double support. This excessive lowering of the CoM means that extra work is done to raise the CoM over the single support leg. The situation is aggravated by the fact that the CoM was lifted higher than typical because of the heel lifting during single support. Although these determinants allow quantification of the effects of gait kinematics and provide some useful information for gait they are limited in their ability to quantify the dynamics and kinetics of gait that are important for individuals with walking disabilities.

Tacrolimus with mycophenolate mofetil (MMF) or sirolimus vs. cyclosporine with MMF in cardiac transplant patients: 1-year report.

The most advantageous combination of immunosuppressive agents for cardiac transplant recipients has not yet been established. Between November 2001 and June 2003, 343 de novo cardiac transplant recipients were randomized to receive steroids and either tacrolimus (TAC) + sirolimus (SRL), TAC + mycophenolate mofetil (MMF) or cyclosporine (CYA) + MMF. Antilymphocyte induction therapy was allowed for up to 5 days. The primary endpoint of >/=3A rejection or hemodynamic compromise rejection requiring treatment showed no significant difference at 6 months (TAC/MMF 22.4%, TAC/SRL 24.3%, CYA/MMF 31.6%, p = 0.271) and 1 year (p = 0.056), but it was significantly lower in the TAC/MMF group when compared only to the CYA/MMF group at 1 year (23.4% vs. 36.8%; p = 0.029). Differences in the incidence of any treated rejection were significant (TAC/SRL = 35%, TAC/MMF = 42%, CYA/MMF = 59%; p < 0.001), as were median levels of serum creatinine (TAC/SRL = 1.5 mg/dL, TAC/MMF = 1.3 mg/dL, CYA/MMF = 1.5 mg/dL; p = 0.032) and triglycerides (TAC/SRL = 162 mg/dL, TAC/MMF = 126 mg/dL, CYA/MMF = 154 mg/dL; p = 0.028). The TAC/SRL group encountered fewer viral infections but more fungal infections and impaired wound healing. These secondary endpoints suggest that the TAC/MMF combination appears to offer more advantages than TAC/SRL or CYA/MMF in cardiac transplant patients, including fewer >/=3A rejections or hemodynamic compromise rejections and an improved side-effect profile.

The benefit of implementing a heart failure disease management program.

BACKGROUND: To handle the increasing complexity of congestive heart failure (CHF) care, several new models for the care of patients with CHF have been developed to replace traditional strategies. We undertook this study to evaluate the potential benefit of implementing a CHF disease management program at a tertiary care center, particularly in terms of beta-blocker use and cost to the health care system. METHODS: After reviewing the literature regarding therapies and management strategies for patients with CHF, we developed the Duke Heart Failure Program. All enrolled patients had 1 of the following: recent CHF hospitalization, ejection fraction less than 20%, or symptoms consistent with New York Heart Association class III or IV. We compared preenrollment and postenrollment medication use and resource utilization. RESULTS: We enrolled 117 patients from July 1998 to April 1999. Mean enrollment time was 4.7 months. beta-Blocker use and dose significantly increased (52% vs 76% for beta-blocker, P<.01; 6% vs 13% of target dose, P<.01). The hospitalization rate decreased (1.5 vs 0 hospitalizations per patient-year, P<.01), while the number of clinic visits increased (4.3 vs 9.8 clinic visits per patient-year, P<.01). The Duke University Health System saved a median of $8571 per patient-year. CONCLUSIONS: Implementing a CHF disease management program was associated with improved CHF medication dosing and with decreased hospitalization for patients with CHF. A CHF disease management program is an effective method for a health care system to care for patients with CHF.

Evaluation of early postoperative results after bicaval versus standard cardiac transplantation and review of the literature.

OBJECTIVE: Previous studies have been inconsistent in defining a clinical benefit to the bicaval cardiac transplantation technique relative to the standard technique, and many major centers have not adopted this newer approach. The purpose of this study was to determine whether clinically significant benefits support utilization of the bicaval technique. METHODS: Sixty-eight consecutive adult patients undergoing a standard cardiac transplant were compared with 75 consecutive patients who underwent the bicaval technique during the period from 1991 to 1999. Etiology, recipient sex, recipient age, donor age, and pulmonary vascular resistance were similar between the two groups. RESULTS: Cardiac index at 24 hours after operation was increased for the bicaval group relative to the standard group (3.15 +/- 0.7 vs 2.7 +/- 0.5 L/min/m(2), P <. 05). Inotropic requirements were significantly less, and there was significantly less tricuspid regurgitation in the bicaval group relative to the standard group. In addition, the bicaval group more frequently had a nonpaced normal sinus rhythm at 24 hours after operation (73.9% vs 50.7% [standard group], P =.025) and had fewer postoperative arrhythmias (29.3% vs 47.7% [standard group], P <.01). Finally, although mortality was similar for the two groups, length of postoperative hospitalization was longer for the standard group relative to the bicaval group (12.1 +/- 11 vs 20.4 +/- 12 days, P <. 001). Review of the literature identified reduced tricuspid regurgitation and improved rhythm as consistent benefits of the bicaval technique. CONCLUSION: This review demonstrates a clinical benefit during the early postoperative period with bicaval cardiac transplantation (relative to standard) and encourages further utilization of this technique.

Bridging patients to cardiac transplantation.

Potential recipients of heart transplants have the most advanced form of congestive heart failure, in which standard therapy fails to maintain clinical stability. In the absence of guidelines derived from evidence obtained in clinical trials, caring for these patients becomes a challenge. A successful approach requires the proper coordination of surgical and nonsurgical strategies, including revascularization and valvular surgery as well as mechanical ventricular support and medical strategies. Intensive medical therapy is the most commonly used approach for prolonged bridging to transplantation. Although carefully individualized regimens are necessary to achieve desired goals, most centers adopt a fairly standardized approach involving vasodilators, diuretics, and inotropic support. Bridging patients with cardiac decompensation to transplantation presents a major therapeutic challenge. Appropriate strategies will maximize patients' chances that the bridge from decompensation to transplantation remains intact. (c)2000 by CHF, Inc.

Lack of effect of increased inspired oxygen concentrations on maximal exercise capacity or ventilation in stable heart failure.

Recent uncontrolled studies have suggested improved maximal exercise capacity and decreased exercise ventilation in heart failure after administration of increased inspired oxygen concentrations. To study the responses further, 16 patients performed staged, symptom-limited cycle ergometry with humidified 21% and 60% inspired oxygen concentrations using a randomized, double-blind, crossover study design. Serial measurements of minute ventilation, heart rate, blood pressure, leg blood flow, and arterial and venous lactate and oxygen content were obtained. Exercise time did not change between the 2 tests (595 +/- 179 seconds and 602 +/- 181 seconds for 21% and 60% oxygen concentrations, respectively). Similarly, measurements of the ventilatory response to exercise and of leg blood flow were not different between the 2 oxygen concentrations. Although hemoglobin oxygen saturation increased from 96.7 +/- 2.1% to 97.9 +/- 1.5% at rest, at both rest and maximal exercise there was no statistically significant difference in arterial or venous oxygen content. This study failed to demonstrate any physiologic or functional benefit from the administration of increased oxygen concentrations to patients with stable heart failure.

Aerobic exercise training can reverse age-related peripheral circulatory changes in healthy older men.

BACKGROUND: The age-related decline in maximal oxygen consumption is attenuated by habitual aerobic exercise. However, the relative effects of training on central and peripheral responses to exercise in older subjects are not known. The present study assessed the contribution of central and peripheral responses to the age-associated decline in peak oxygen consumption and compared the effect of exercise training in healthy older and younger subjects. METHODS AND RESULTS: Ten older and 13 younger men underwent invasive measurement of central and peripheral cardiovascular responses during an upright, staged cycle exercise test before and after a 3-month period of exercise training with cycle ergometry. At baseline, cardiac output and AV oxygen difference during exercise were significantly lower in older subjects. With training, the older and younger groups increased maximal oxygen consumption by 17.8% and 20.2%, respectively. Peak cardiac output was unchanged in both groups. Systemic AV oxygen difference increased 14.4% in the older group and 14.3% in the younger group and accounted for changes in peak oxygen consumption. Peak leg blood flow increased by 50% in older subjects, whereas the younger group showed no significant change. There was no change in peak leg oxygen extraction in the older group, but in the younger group, leg AV oxygen difference increased by 15.4%. CONCLUSIONS: These findings suggest that the age-related decline in maximal oxygen consumption results from a reversible deconditioning effect on the distribution of cardiac output to exercising muscle and an age-related reduction in cardiac output reserve.

Localization of the Ca(2+)-binding protein, Bra r 1, in anthers and pollen tubes.

Calcium plays an essential role during pollen development and pollen tube growth, and several Ca(2+)-binding proteins are expressed in anthers. We have previously reported that Brassica pollen allergens encoded by Bra r 1 and Bra r 2 show sequence similarities to Ca(2+)-binding proteins [Toriyama et al. (1995) Plant Mol. Biol. 29: 1157]. Herein, we report that both genes are expressed in the diploid tapetum and haploid microspores, as detected by in situ RNA hybridization. Immunoblot analysis revealed that Bra r 1 and Bra r 2 were accumulated in anthers during pollen development. When pollen grains were suspended in an aqueous solution, both proteins were mainly detected in the pollen extracellular fraction, indicating that Bra r 1 and Bra r 2 are released from the pollen upon hydration. Localization of Bra r 1 was further investigated in sections of anthers and pollen tubes. Bra r 1 was detected in the tapetum, microspores and pollen grains. In longitudinal sections of cross-pollinated pistils. Bra r 1 was detected throughout pollen tubes elongating in transmitting-tissue. These findings suggest that Bra r 1 may be involved in pollen-pistil interaction and pollen tube growth.

Exertional dyspnea in heart failure: a symptom unrelated to pulmonary function at rest or during exercise. Duke University Clinical Cardiology Studies (DUCCS) Exercise Group.

BACKGROUND: Exertional dyspnea is a common symptom in patients with heart failure, and the mechanisms responsible for the symptom are unknown. The purpose of this study was to identify factors responsible for the symptom of exertional dyspnea in patients with heart failure. METHODS: Resting pulmonary-function tests and maximal cardiopulmonary exercise tests were performed in 71 patients with New York Heart Association functional class II-IV symptoms (mean ejection fraction 30.6%; mean age, 68 years). RESULTS: The severity of dyspnea at peak exercise, which patients rated as 3 to 10 on a 1 to 10 severity scale, did not correlate with rest or exercise hemodynamic, spirometric, or metabolic variables, including peak oxygen uptake (VO2), minute ventilation (Ve), and respiratory rate, or with derived variables including Ve/VO2, Ve/VCO2, and the dyspnea index (Ve/maximum voluntary ventilation). Additionally, these variables did not differ between patients who reported limitation of exercise by dyspnea and those who were limited by fatigue. CONCLUSIONS: The symptom of exertional dyspnea in patients with heart failure is not determined by abnormalities in ventilatory function or demand.

Improvement in the mechanical efficiency of walking: an explanation for the "placebo effect" seen during repeated exercise testing of patients with heart failure. Duke University Clinical Cardiology Studies (DUCCS) Exercise Group.

To determine the mechanism responsible for the "placebo effect" seen during serial exercise testing of patients with heart failure, we examined metabolic variables for 81 patients who underwent five baseline exercise tests as part of a multicenter drug trial. The patients were 50 men and 31 women with a mean ejection fraction of 30.1% and a mean age of 69 years. From test 1 to 2, the exercise time increased from 419 +/- 140 to 462 +/- 130 seconds before it reached a plateau over the next three tests. Metabolic measurements at test 1 and test 3 revealed no change in peak oxygen consumption ( 1119 +/- 376 to 1105 +/- 346 ml/min). Maximum heart rate, systolic blood pressure, ventilation, and respiratory exchange ratio also were unchanged. The onset of the anaerobic threshold was delayed from 211 +/- 81 to 238 +/- 93 seconds, but there was no change in oxygen consumption at the anaerobic threshold (810 +/- 222 to 795 +/- 220 ml/min). At a predetermined submaximal level, oxygen consumption, ventilation, and respiratory exchange ratio all decreased to a statistically significant degree. These results indicate that a rapid increase in the mechanical efficiency of walking contributes to the placebo effect among patients with heart failure during serial exercise testing and is independent of changes in conditioning or motivation.

Isolation and collection of two populations of viable sperm cells from the pollen of Plumbago zeylanica.

A protocol is described for individually collecting two populations of sperm cells, Svn and Sua, from pollen of Plumbago zeylanica. Pollen grains were burst in 10 mM MOPS buffer containing 0.8 M mannitol (pH 4.6). Paired sperm cells released from pollen were separated using a microinjector. Svn and Sua were then collected individually with a microinjector, based upon known size differences. Collected sperm cells were washed with isolation medium and transferred to liquid nitrogen until use. Fluorochromatic reaction (FCR) test of isolated sperm cells showed a positive reaction, indicating that the isolated sperm cells are viable; most of the sperm cells retain viability for at least 2 h.

Spray-freezing freeze substitution (SFFS) of cell suspensions for improved preservation of ultrastructure.

Some unicellular organisms present challenges to chemical fixations that lead to common, yet obvious, artifacts. These can be avoided in entirety by adapting spray-freezing technology to ultrarapidly freeze specimens for freeze substitution. To freeze specimens, concentrated suspensions of cells ranging in diameter from 0.5-30 pm were sprayed with an airbrush at 140-200 kPa (1.05-1.5 torr; 20.3-29.0 psi) into a nylon mesh transfer basket submerged in liquid propane. After freezing, the mesh basket containing the frozen sample was lifted out of the chamber, drained and transferred through several anhydrous acetone rinses at 188 K (-85 degrees C). Freeze substitution was conducted in 1% tannic acid/1% anhydrous glutaraldehyde in acetone at 188 K (-85 degrees C), followed by 1% OsO4/acetone at 277 K (4 degrees C). Freeze substitution was facilitated using a shaking table to provide gentle mixing of the substitution medium on dry ice. High quality freezing was observed in 70% of spray-frozen dinoflagellate cells and in 95% of spray-frozen cyanobacterial cells. These could be infiltrated and observed directly; however, overall ultrastructural appearance and membrane contrast were improved when the freeze-substituted cells were rehydrated and post-fixed in aqueous OSO4, then dehydrated and embedded in either Spurr's or Epon resin. Ultrastructural preservation using this ultrarapid freezing method provided specimens that were consistently superior to those obtainable in even the best comparable chemical fixations.

Micromanipulation of male and female gametes ofNicotiana tabacum: I. Isolation of gametes.

The isolation of male and female gametes is a precondition for the micromanipulation of flowering plant gametes. To reflect their condition at fertilization, isolated gametes need to be physiologically mature and vigorous. Sperm cells are isolated from pollen tubes grown on cut styles using the "in vivo/in vitro" technique. Embryo sacs are isolated 2 days after anthesis using brief treatments of minimal concentrations of cell-wall-digesting enzymes on ovules of emasculated flowers. Egg cells are then mechanically separated from the embryo sac, allowing unambiguous identification of cells. Two days is usually the minimum required for the pollen tube to penetrate the ovule and effect fertilization in vivo.

Micromanipulation of male and female gametes ofNicotiana tabacum: II. Preliminary attempts for in vitro fertilization and egg cell culture.

This research is part of an attempt to establish an in vitro fertilization system in tobacco to aid in understanding mechanisms of fertilization. Fusions of isolated male and female gametes were induced in a polyethylene glycol solution. Fusion appears similar to that in maize. One nuclear division of both an unfertilized egg cell and a synergid was induced in KM8p medium with 1 mg/l 2,4-dichlorophenoxyacetic acid in a microchamber culture; one cellular division of the egg cell was also induced in the same medium in solid-drop culture. The osmolality of suspension culture feeder cells was critical for the development of these cells. These results indicate that in vitro fertilization is possible in tobacco, which would be the first such system in dicots.

Isolation of viable sperm cells from tobacco (Nicotiana tabacum).

Viable sperm cells of Nicotiana tabacum were isolated by the semi-vivo technique. After pollination, excised styles were floated, cut end immersed, in a solution of 15% sucrose with 0.01% boric acid and 0.03% Ca(NO3)2 at 27 degrees C in a growth chamber until pollen tubes emerged. After sperm cells were formed (at least 8 h after pollination) tubes were immersed in a 9% mannitol solution. In this solution, sperm cells are nearly ellipsoidal and retain viability for over 6 h.

Occurrence of Mitochondria in the Nuclei of Tobacco Sperm Cells.

Tobacco sperm cells contain intact mitochondria within their nuclei with a frequency of 0.35 [plusmn] 0.13 per cell. These inclusions appear to originate from mitochondria found among chromatids in the highly elongated metaphase plate of the dividing generative cell. These organelles are apparently captured during the reconstitution of the nuclear envelope. Only sperm cells were observed to contain these nuclear mitochondria; generative cells, vegetative pollen cells, transmitting tissue cells, unfertilized egg cells, and central cells lacked them. Nuclear mitochondria were also seen in the nuclei of the egg and central cell after fusion with sperm nuclei, suggesting that nuclear mitochondria are transmitted into the zygote and primary endosperm cells during double fertilization. Organellar inclusions in the sperm nucleus provide a potential mechanism for transmitting organellar DNA into the next generation and could potentially facilitate the transfer of genetic material between the nucleus and other organelles.