Advances in understanding the energetics of muscle contraction.

Muscle energetics encompasses the relationships between mechanical performance and the biochemical and thermal changes that occur during muscular activity. The biochemical reactions that underpin contraction are described and the way in which these are manifest in experimental recordings, as initial and recovery heat, is illustrated. Energy use during contraction can be partitioned into that related to cross-bridge force generation and that associated with activation by Ca2+. Activation processes account for 25-45% of ATP turnover in an isometric contraction, varying amongst muscles. Muscle energy use during contraction depends on the nature of the contraction. When shortening muscles produce less force than when contracting isometrically but use energy at a greater rate. These characteristics reflect more rapid cross-bridge cycling when shortening. When lengthening, muscles produce more force than in an isometric contraction but use energy at a lower rate. In that case, cross-bridges cycle but via a pathway in which ATP splitting is not completed. Shortening muscles convert part of the free energy available from ATP hydrolysis into work with the remainder appearing as heat. In the most efficient muscle studied, that of a tortoise, cross-bridges convert a maximum of 47% of the available energy into work. In most other muscles, only 20-30% of the free energy from ATP hydrolysis is converted into work.

The energetics of muscle contractions resembling in vivo performance.

Muscle energetics has expanded into the study of contractions that resemble in vivo muscle activity. A summary is provided of experiments of this type and what they have added to our understanding of muscle function and effects of compliant tendons, as well as the new questions raised about the efficiency of energy transduction in muscle.

A century of exercise physiology: key concepts in muscle energetics.

In the mid-nineteenth century, the concept of muscle behaving like a stretched spring was developed. This elastic model of contraction predicted that the energy available to perform work was established at the start of a contraction. Despite several studies showing evidence inconsistent with the elastic model, it persisted into the twentieth century. In 1923, W. O. Fenn published a paper in which he presented evidence that appeared to clearly refute the elastic model. Fenn showed that when a muscle performs work it produces more heat than when contracting isometrically. He proposed that energy for performing work was only made available in a muscle as and when that work was performed. However, his ideas were not adopted and it was only after 15 years of technical developments that in 1938 A. V. Hill performed experiments that conclusively disproved the elastic model and supported Fenn's conclusions. Hill showed that the rate of heat production increased as a muscle made the transition from isometric to working contraction. Understanding the basis of the phenomenon observed by Fenn and Hill required another 40 years in which the processes that generate force and work in muscle and the associated scheme of biochemical reactions were established. Demonstration of the biochemical equivalent of Hill's observations-changes in rate of ATP splitting when performing work-in 1999 was possible through further technical advances. The concept that the energy, from ATP splitting, required to perform work is dynamically modulated in accord with the loads a muscle encounters when contracting is key to understanding muscle energetics.

Historical Perspective: Heat production and chemical change in muscle. Roger C. Woledge.

The objective of this article is to provide an historical perspective on a review of "Heat production and chemical change in muscle" written by Roger C. Woledge and published in Progress in Biophysics and Molecular Biology 50 years ago. We first provide a brief but broad summary of the history of muscle chemistry prior to 1971 and then address the central theme of the 1971 review - that of energy balance. Energy balance is a method to establish whether all the energetically significant biochemical reactions accompanying muscle contraction have been identified. Woledge adopted the method to compare the measured enthalpy output (i.e., the sum of the heat output and work output) to that expected from the extent of known biochemical reactions. Prior work had suggested that the observed and expected enthalpy outputs were similar but Woledge proposed that the expected heat had been overestimated and that, hence, there must be an unidentified reaction that accounted for as much as half the heat produced by a contracting muscle. We describe investigations carried out after the review that vindicated that view, ultimately characterising the processes producing the unexplained enthalpy which, in turn, led to identification of the hitherto unknown reaction. Those experiments and a more recent resurrection of the approach using fluorescent probes to monitor ATP turnover have now accounted for the processes that underlie the complex time courses of muscle heat production and ATP turnover during contraction, at least in the classical frog sartorius muscle preparation. However, the few studies performed on mammalian muscles since then have produced results that are difficult to reconcile with the ideas derived from energy balance studies of amphibian and fish muscles, thereby suggesting a new objective for energy balance studies.

Components of activation heat in skeletal muscle.

Activation heat (qA) production by muscle is the thermal accompaniment of the release of Ca2+ from the sarcoplasmic reticulum (SR) into the cytoplasm, its interactions with regulatory proteins and other cytoplasmic Ca2+ buffers and its return to the SR. The contribution of different Ca2+-related reactions to qA is difficult to determine empirically and therefore, for this study, a mathematical model was developed to describe Ca2+ movements and accompanying thermal changes in muscle fibres in response to stimulation. The major sources of heat within a few milliseconds of the initiation of Ca2+ release are Ca2+ binding to Tn and Pv. Ca2+ binding to ATP produces a relatively small amount of heat. Ca2+ dissociation from ATP and Tn, with heat absorption, are of similar time course to the decline of force. In muscle lacking Pv (e.g. mouse soleus), Ca2+ is then rapidly pumped into the SR. In muscles with Pv, Ca2+ that dissociates from Tn and ATP binds to Pv and then dissociates slowly (over 10 s of seconds) and is then pumped into the SR; the net effect of these two processes is heat absorption. It is proposed that this underlies Hill's "negative delayed heat". After all the Ca2+ is returned to the SR, qA is proportional to the amount of Ca2+ released into the cytoplasm. In muscles with Pv this is 20-60 s after Ca2+ release; in muscles without Pv, all Ca2+ is returned to the SR soon after the end of force relaxation.

Extended Treatment with Glial Cell Line-Derived Neurotrophic Factor in Parkinson's Disease.

BACKGROUND: Intraputamenal glial cell line-derived neurotrophic factor (GDNF), administered every 4 weeks to patients with moderately advanced Parkinson's disease, did not show significant clinical improvements against placebo at 40 weeks, although it significantly increased [18F]DOPA uptake throughout the entire putamen. OBJECTIVE: This open-label extension study explored the effects of continued (prior GDNF patients) or new (prior placebo patients) exposure to GDNF for another 40 weeks. METHODS: Using the infusion protocol of the parent study, all patients received GDNF without disclosing prior treatment allocations (GDNF or placebo). The primary outcome was the percentage change from baseline to Week 80 in the OFF state Unified Parkinson's Disease Rating Scale (UPDRS) motor score. RESULTS: All 41 parent study participants were enrolled. The primary outcome decreased by 26.7±20.7% in patients on GDNF for 80 weeks (GDNF/GDNF; N = 21) and 27.6±23.6% in patients on placebo for 40 weeks followed by GDNF for 40 weeks (placebo/GDNF, N = 20; least squares mean difference: 0.4%, 95% CI: -13.9, 14.6, p = 0.96). Secondary endpoints did not show significant differences between the groups at Week 80 either. Prespecified comparisons between GDNF/GDNF at Week 80 and placebo/GDNF at Week 40 showed significant differences for mean OFF state UPDRS motor (-9.6±6.7 vs. -3.8±4.2 points, p = 0.0108) and activities of daily living score (-6.9±5.5 vs. -1.0±3.7 points, p = 0.0003). No treatment-emergent safety concerns were identified. CONCLUSIONS: The aggregate study results, from the parent and open-label extension suggest that future testing with GDNF will likely require an 80- rather than a 40-week randomized treatment period and/or a higher dose.

Randomized trial of intermittent intraputamenal glial cell line-derived neurotrophic factor in Parkinson's disease.

We investigated the effects of glial cell line-derived neurotrophic factor (GDNF) in Parkinson's disease, using intermittent intraputamenal convection-enhanced delivery via a skull-mounted transcutaneous port as a novel administration paradigm to potentially afford putamen-wide therapeutic delivery. This was a single-centre, randomized, double-blind, placebo-controlled trial. Patients were 35-75 years old, had motor symptoms for 5 or more years, and presented with moderate disease severity in the OFF state [Hoehn and Yahr stage 2-3 and Unified Parkinson's Disease Rating Scale motor score (part III) (UPDRS-III) between 25 and 45] and motor fluctuations. Drug delivery devices were implanted and putamenal volume coverage was required to exceed a predefined threshold at a test infusion prior to randomization. Six pilot stage patients (randomization 2:1) and 35 primary stage patients (randomization 1:1) received bilateral intraputamenal infusions of GDNF (120 µg per putamen) or placebo every 4 weeks for 40 weeks. Efficacy analyses were based on the intention-to-treat principle and included all patients randomized. The primary outcome was the percentage change from baseline to Week 40 in the OFF state (UPDRS-III). The primary analysis was limited to primary stage patients, while further analyses included all patients from both study stages. The mean OFF state UPDRS motor score decreased by 17.3 ± 17.6% in the active group and 11.8 ± 15.8% in the placebo group (least squares mean difference: -4.9%, 95% CI: -16.9, 7.1, P = 0.41). Secondary endpoints did not show significant differences between the groups either. A post hoc analysis found nine (43%) patients in the active group but no placebo patients with a large clinically important motor improvement (≥10 points) in the OFF state (P = 0.0008). 18F-DOPA PET imaging demonstrated a significantly increased uptake throughout the putamen only in the active group, ranging from 25% (left anterior putamen; P = 0.0009) to 100% (both posterior putamina; P < 0.0001). GDNF appeared to be well tolerated and safe, and no drug-related serious adverse events were reported. The study did not meet its primary endpoint. 18F-DOPA imaging, however, suggested that intermittent convection-enhanced delivery of GDNF produced a putamen-wide tissue engagement effect, overcoming prior delivery limitations. Potential reasons for not proving clinical benefit at 40 weeks are discussed.

Restorative aspects of oral cancer reconstruction.

The oral rehabilitation of head and neck cancer patients is a vital and pivotal part of their care pathway, helping to restore function, aesthetics and improve patient well-being and quality of life. Patient-reported outcome data demonstrates that such patients consider that many of their continued and unmet needs relate to their oral and dental rehabilitation. It is therefore incumbent on all dental team members to be aware of the entire spectrum of restorative management options that can be utilised in both the prevention and rehabilitation of this patient cohort. The literature highlights that maxillary and mid-face tumours are challenging, as patients often suffer both profound functional and aesthetic consequences with significant physiological and psychological impact. Conversely, mandibular and floor of mouth tumours often result in issues primarily affecting oral function. This paper will summarise commonly encountered functional and aesthetic issues facing the restorative dental team and highlight widespread preventive challenges.

Mini Dental Implants in the Management of The Atrophic Maxilla and Mandible: A New Implant Design and Preliminary Results.

Although the edentulous population in the UK is falling, those that are rendered edentulous are becoming edentate later in life and with significantly resorbed ridges. This creates a challenge because the management of such patients and their ability to adapt to new dentures is impaired later in life. Despite widespread endorsement of two implants to retain lower complete dentures, the inability to comply has resulted in elderly patients with compromised ability to function and unable to eat a healthy diet. Mini dental implants may offer an ideal solution for the elderly edentulous population who may not be keen on invasive surgery for the placement of conventional dental implants. Further work is required to show the longevity of these restorations, however, existing research and clinical experience show that they potentially offer a simple solution to this group of patients. This paper presents the development of a new design of mini implant, based on clinical problems encountered during a pilot randomised controlled trial. The design of the new implant specifically aims to overcome problems in managing severely atrophic ridges. A preliminary survival study shows survival rates to be equivalent to other mini dental implants and highly satisfactory in the short to medium term.

The use of dental implants, cast bars and sleeve overdentures in oral cancer patients.

Surgical resection of an oral tumour (and the associated free flap reconstruction) can significantly alter the oral anatomy. The lack of sulcus depth, alveolar ridge, presence of a mobile flap and limited tongue movement can make it impossible for patients to control a removable prosthesis. To help this cohort of patients, dental implants can be invaluable. The Oral Rehabilitation Team at Central Manchester University Dental Hospital have used dental implants to rehabilitate oral cancer patients for over thirty years. After their resective surgery, a number of patients were dentally rehabilitated with a laboratory-made, precious metal-alloy bar supported by at least four dental implants. A metal-alloy under-sleeve retained overdenture was then provided to fit over the milled bar. The majority of the 50 patients in this case series had tumours in the anterior floor of the mouth. It was noted that 76% of the patients received a rim resection and were reconstructed with a fasciocutaneous, soft tissue free flap. Six percent of patients received a segmental resection and were reconstructed with either a fibular or deep circumflex iliac artery free flap. The dental implants and sleeve overdentures had a survival rate of 100%. None of the dentures lost retention, implying that the frictional grip between the overcasting and the milled bar was sufficient to appease the retention demands of this cohort. However, 10% of patients encountered complications. This would suggest a success (or complication free) rate of 90% for this cohort of 50 oncology patients. This would still imply that milled bars/sleeve overdentures carry a relatively low maintenance burden and may be a useful treatment option for oral cancer patients.

Energy demand and supply in human skeletal muscle.

The energy required for muscle contraction is provided by the breakdown of ATP but the amount of ATP in muscles cells is sufficient to power only a short duration of contraction. Buffering of ATP by phosphocreatine, a reaction catalysed by creatine kinase, extends the duration of activity possible but sustained activity depends on continual regeneration of PCr. This is achieved using ATP generated by oxidative processes and, during intense activity, by anaerobic glycolysis. The rate of ATP breakdown ranges from 70 to 140 mM min-1 during isometric contractions of various intensity to as much as 400 mM min-1 during intense, dynamic activity. The maximum rate of oxidative energy supply in untrained people is ~50 mM min-1 which, if the contraction duty cycle is 0.5 as is often the case in cyclic activity, is sufficient to match an ATP breakdown rate during contraction of 100 mM min-1. During brief, intense activity the rate of ATP turnover can exceed the rates of PCr regeneration by combined oxidative and glycolytic energy supply, resulting in a net decrease in PCr concentration. Glycolysis has the capacity to produce between 30 and 50 mM of ATP so that, for example, anaerobic glycolysis could provide ATP at an average of 100 mM min-1 over 30 s of exhausting activity. The creatine kinase reaction plays an important role not only in buffering ATP but also in communicating energy demand from sites of ATP breakdown to the mitochondria. In that role, creatine kinases acts to slow and attenuate the response of mitochondria to changes in energy demand.

Restorative dentistry and oral rehabilitation: United Kingdom National Multidisciplinary Guidelines.

This is the official guideline endorsed by the specialty associations involved in the care of head and neck cancer patients in the UK and provides recommendations on the pre-treatment oral and dental assessment, during and after treatment and oral rehabilitation. Restorative dentists are core members of the multidisciplinary team treating head and neck cancer patients, involved from the treatment planning phase through to long-term rehabilitation. Recommendations • Preventative oral care must be delivered to patients whose cancer treatment will affect the oral cavity, jaws, salivary glands and oral accessibility. (G) • Close working and communication between the surgeons, oncologists and restorative dental specialists is important in ensuring optimal oral health outcomes. (G) • Intensity-modulated radiotherapy has been shown to reduce long-term xerostomia and should be offered to all appropriate patients. (R) • If patients are deemed at risk of trismus they should be warned and its progressive and potentially irreversible nature explained. (G) • Where it is known that adjuvant radiotherapy will be given, extractions should take place at primary surgery to maximise the time for healing and minimise the number of surgical events for patients. (G) • Osseointegrated implants should be considered for all patients having resection for head and neck cancer. (G).

A mathematical model of heat flow in a thermopile for measuring muscle heat production: implications for design and signal analysis.

Contracting muscles produce heat which largely arises from the biochemical reactions that provide the energy for contraction. Measurements of muscle heat production have made, and continue to make, important contributions to our understanding of the bases of contraction. Most measurements of muscle heat production are made using a thermopile, consisting of a series of thermocouples arranged so that alternate thermocouples are in thermal contact with the muscle and with an isothermal reference. In this study, a mathematical model was constructed of a muscle lying on a thermopile consisting of antimony-bismuth thermocouples sandwiched between polymer sheets. The validity of the model was demonstrated by its ability to accurately predict thermopile outputs in response to applying heat to the thermopile surface, to generating heat in the thermocouples using the Peltier effect and to adding heat capacity on the thermopile surface. The model was then used to show how practical changes to thermopile construction could minimise response time and thermopile heat capacity and allow measurement of very low rates of heat production. The impulse response of a muscle-thermopile system was generated using the model and used to illustrate how a measured signal can be deconvolved with the impulse response to correct for lag introduced by the thermopile.

Energetics of contraction.

Muscles convert energy from ATP into useful work, which can be used to move limbs and to transport ions across membranes. The energy not converted into work appears as heat. At the start of contraction heat is also produced when Ca(2+) binds to troponin-C and to parvalbumin. Muscles use ATP throughout an isometric contraction at a rate that depends on duration of stimulation, muscle type, temperature and muscle length. Between 30% and 40% of the ATP used during isometric contraction fuels the pumping Ca(2+) and Na(+) out of the myoplasm. When shortening, muscles produce less force than in an isometric contraction but use ATP at a higher rate and when lengthening force output is higher than the isometric force but rate of ATP splitting is lower. Efficiency quantifies the fraction of the energy provided by ATP that is converted into external work. Each ATP molecule provides 100 zJ of energy that can potentially be converted into work. The mechanics of the myosin cross-bridge are such that at most 50 zJ of work can be done in one ATP consuming cycle; that is, the maximum efficiency of a cross-bridge is ∼50%. Cross-bridges in tortoise muscle approach this limit, producing over 90% of the possible work per cycle. Other muscles are less efficient but contract more rapidly and produce more power.

Development and performance of electronic acute kidney injury triggers to identify pediatric patients at risk for nephrotoxic medication-associated harm.

BACKGROUND: Nephrotoxic medication-associated acute kidney injury (NTMx-AKI) is a costly clinical phenomenon and more common than previously recognized. Prior efforts to use technology to identify AKI have focused on detection after renal injury has occurred. OBJECTIVES: Describe an approach and provide a technical framework for the creation of risk-stratifying AKI triggers and the development of an application to manage the AKI trigger data. Report the performance characteristics of those triggers and the refinement process and on the challenges of implementation. METHODS: Initial manual trigger screening guided design of an automated electronic trigger report. A web-based application was designed to alleviate inefficiency and serve as a user interface and central workspace for the project. Performance of the NTMx exposure trigger reports from September 2011 to September 2013 were evaluated using sensitivity (SN), specificity (SP), positive and negative predictive values (PPV, NPV). RESULTS: Automated reports were created to replace manual screening for NTMx-AKI. The initial performance of the NTMx exposure triggers for SN, SP, PPV, and NPV all were ≥0.78, and increased over the study, with all four measures reaching ≥0.95 consistently. A web-based application was implemented that simplifies data entry and couriering from the reports, expedites results viewing, and interfaces with an automated data visualization tool. Sociotechnical challenges were logged and reported. CONCLUSION: We have built a risk-stratifying system based on electronic triggers that detects patients at-risk for NTMx-AKI before injury occurs. The performance of the NTMx-exposed reports has neared 100% through iterative optimization. The complexity of the trigger logic and clinical workflows surrounding NTMx-AKI led to a challenging implementation, but one that has been successful from technical, clinical, and quality improvement standpoints. This report summarizes the construction of a trigger-based application, the performance of the triggers, and the challenges uncovered during the design, build, and implementation of the system.

Efficiency and cross-bridge work output of skeletal muscle is decreased at low levels of activation.

The purpose of this study was to determine how the mechanical efficiency of skeletal muscle is affected by level of activation. Experiments were performed in vitro (35 °C) using bundles of fibres from fast-twitch extensor digitorum longus (EDL) and slow-twitch soleus muscles of mice. Measurements were made of the total work and heat produced in response to 10 brief contractions. Mechanical efficiency was the ratio of total work performed to (total heat produced + work performed). Level of activation was varied by altering stimulation frequency between 40 and 160 Hz. Efficiency did not differ significantly between the two muscle types but was significantly lower using 40 Hz stimulation (mean efficiency ± SEM, 0.092 ± 0.012, n = 12, averaged across EDL and soleus) than at any of the other frequencies (160 Hz: 0.147 ± 0.007, n = 12). Measurements of the partitioning of energy output between force-dependent and force-independent components enabled calculation of the amount of Ca(2+) released and number of cross-bridge cycles performed during the contractions. At 40 Hz stimulation frequency, less Ca(2+) was released than at higher frequencies and fewer cross-bridge cycles were performed. Furthermore, less work was performed in each cross-bridge cycle. It is concluded that skeletal muscles are less efficient at low levels of activation than when fully activated and this indicates that level of activation affects not only the number of cycling cross-bridges but also the ability of individual cross-bridges to perform work.

Can mobile technology improve response times of junior doctors to urgent out-of-hours calls? A prospective observational study.

BACKGROUND: The Hospital at Night system has been widely adopted to manage Out-of-Hours workload. However, it has the potential to introduce delays and corruption of information. The introduction of newer technologies to replace landlines, pagers and paper may ameliorate these issues. AIM: To establish if the introduction of a Hospital at Night system supported by a wireless taskflow system affected the escalation of high Early Warning Scores (EWSs) to medical attention, and the time taken to medical review. DESIGN: Prospective 'pre and post' observational study in a teaching hospital in the UK. METHODS: Review of observation charts and medical records, and data extraction from the electronic taskflow system. RESULTS: The implementation of a technology-supported Hospital at Night system was associated with a significant decrease in time to documentation of initial review in those who were reviewed. However, there was no change in the proportion of those with a high EWS that were reviewed, and throughout the study a majority of patients with high EWSs were not reviewed in accordance with guidelines. CONCLUSION: Introduction of a Hospital at Night system supported by mobile technology appeared to improve the transfer of information, but did not affect the nursing decision whether to escalate abnormal findings.

Quantifying Ca2+ release and inactivation of Ca2+ release in fast- and slow-twitch muscles.

The aims of this study were to quantify the Ca(2+) release underlying twitch contractions of mammalian fast- and slow-twitch muscle and to comprehensively describe the transient inactivation of Ca(2+) release following a stimulus. Experiments were performed using bundles of fibres from mouse extensor digitorum longus (EDL) and soleus muscles. Ca(2+) release was quantified from the amount of ATP used to remove Ca(2+) from the myoplasm following stimulation. ATP turnover by crossbridges was blocked pharmacologically (N-benzyl-p-toluenesulphonamide for EDL, blebbistatin for soleus) and muscle heat production was used as an index of Ca(2+) pump ATP turnover. At 20°C, Ca(2+) release in response to a single stimulus was 34 and 84 µmol (kg muscle)(-1) for soleus and EDL, respectively, and increased with temperature (30°C: soleus, 61 µmol kg(-1); EDL, 168 µmol kg(-1)). Delivery of another stimulus within 100 ms of the first produced a smaller Ca(2+) release. The maximum magnitude of the decrease in Ca(2+) release was greater in EDL than soleus. Ca(2+) release recovered with an exponential time course which was faster in EDL (mean time constant at 20°C, 32.1 ms) than soleus (65.6 ms) and faster at 30°C than at 20°C. The amounts of Ca(2+) released and crossbridge cycles performed are consistent with a scheme in which Ca(2+) binding to troponin-C allowed an average of ∼1.7 crossbridge cycles in the two muscles.

A compliant tendon increases fatigue resistance and net efficiency during fatiguing cyclic contractions of mouse soleus muscle.

AIM: As muscles fatigue, their ability to generate mechanical work decreases as a result of decreased force generation and in cyclic activity, slower the relaxation. The purpose of this study was to determine whether a compliant tendon, connected in series with a muscle, would increase sustained work output during cyclic contractions. METHODS: Experiments were performed in vitro (37 °C) using fibre bundles from mouse soleus muscles (n = 7). Each muscle performed two series of 40 brief contractions at a contraction frequency of 2 Hz and with a sinusoidal length change. One series was performed using the fibre bundle only and one with the fibre bundle and a compliant strip of latex connected between the muscle and the force recording apparatus. RESULTS: When contracting with the latex strip, muscle work output was better maintained during the second half of the protocol than when performed without the latex, overall energy cost was reduced and mechanical efficiency was increased. CONCLUSION: The provision of a compliant tendon analogue increased the level of work output that could be sustained during cyclic contractions and reduced energy expenditure. It is proposed that both metabolic and mechanical consequences of the compliant tendon contribute to the improved performance.

First Report of Cylindrocladiella parva as a Grapevine Pathogen in New Zealand.

Isolates morphologically identified as Cylindrocladiella parva were isolated from characteristic black foot symptoms on a grapevine (Vitis vinifera) rooted on 101-14 rootstock from Central Otago in 2005 and 101-14 rootstocks from a nursery in the Auckland Region in 2007 and 2008. On potato dextrose agar, the isolates initially produced cottony, white mycelia that turned grayish cream or golden cream within 10 days, the initially tawny colony undersides becoming dark brown with age. Conidia (0 to 1 septate; 16.4 to 17.0 [16.7] × 2.3 to 2.6 [2.5] µm) and abundant chlamydospores were produced. To confirm identity of the isolates, genomic DNA was extracted and the ribosomal DNA (rDNA) and ß-tubulin gene were amplified and sequenced (3,4). Sequences of the PCR products were compared with sequences in GenBank. The rDNA (535 bp) and ß-tubulin (297 bp) sequences of the four isolates were 100 and 99% identical, respectively, to reported sequences of C. parva in GenBank (AY793454, grapevine isolate (4)/AY793455 for rDNA; AY793486/AY793488, grapevine isolate (4)/AY793489/HM034822 for ß-tubulin). Although C. parva was previously isolated from grapevines in New Zealand (2) and rootstocks of mature grapevines, cuttings, and graft unions of grafted young grapevines in South Africa (4), its role as a pathogen of Vitis spp. has not been confirmed (2,4). However, it has been reported as a pathogen of Eucalyptus spp. (1) and was also isolated from Telopea speciosissima and Macadamia integrifolia in New Zealand (2,4). The C. parva isolates were tested as a mixed inoculum (four isolates) for pathogenicity on roots of 10 grapevine rootstock plants each of cvs. 101-14 and Schwarzmann (Sch). The rootstocks were grown in potting mix for 4 months, after which the root systems of all vines were wounded with an asparagus knife with a sharp, square tip, driven vertically down into the soil at four equidistant locations approximately 8 cm from the trunk. Each plant was inoculated with 50 ml of the mixed-isolate conidial suspension (106/ml), or 50 ml water (controls), followed by 50 ml of water. After 7 months of growth, the plants were harvested. For C. parva-inoculated plants, internal blackening of the stem base tissue was observed. Isolations from surface-sterilized trunk bases recovered C. parva from four and nine plants of 101-14 and Sch, respectively, with C. parva infections in 25 and 48%, respectively, of the four wood pieces taken per plant. Plants inoculated with water had no blackening and no C. parva was isolated from their stem bases. Mean shoot dry weights of inoculated plants (17.9 and 15.0 g for 101-14 and Sch, respectively) were significantly lower (P = 0.035) than noninoculated controls (26.5 and 20.0 g for 101-14 and Sch, respectively). Mean root dry weights were reduced by C. parva inoculation, although not significantly (32.7 and 27.0 g for C. parva inoculated 101-14 and Sch, respectively, and 36.2 and 27.4 g for control 101-14 and Sch, respectively). To our knowledge, this is the first report of C. parva as a pathogen of grapevines (2,4) and suggests that along with Cylindrocarpon spp., C. parva is part of the pathogen complex responsible for black foot of grapevines. References: (1) P. W. Crous et al. Plant Pathol. 42:302, 1993. (2) P. D. Gadgil et al. Fungi on Trees and Shrubs in New Zealand. Fungal Diversity Press, Hong Kong, 2005. (3) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (4) G. J. van Coller et al. Australas. Plant Pathol. 34:489, 2005.