Interlaboratory evaluation of Mucorales PCR assays for testing serum specimens: A study by the fungal PCR Initiative and the Modimucor study group.

Interlaboratory evaluations of Mucorales qPCR assays were developed to assess the reproducibility and performance of methods currently used. The participants comprised 12 laboratories from French university hospitals (nine of them participating in the Modimucor study) and 11 laboratories participating in the Fungal PCR Initiative. For panel 1, three sera were each spiked with DNA from three different species (Rhizomucor pusillus, Lichtheimia corymbifera, Rhizopus oryzae). For panel 2, six sera with three concentrations of R. pusillus and L. corymbifera (1, 10, and 100 genomes/ml) were prepared. Each panel included a blind negative-control serum. A form was distributed with each panel to collect results and required technical information, including DNA extraction method, sample volume used, DNA elution volume, qPCR method, qPCR template input volume, qPCR total reaction volume, qPCR platform, and qPCR reagents used. For panel 1, assessing 18 different protocols, qualitative results (positive or negative) were correct in 97% of cases (70/72). A very low interlaboratory variability in Cq values (SD = 1.89 cycles) were observed. For panel 2 assessing 26 different protocols, the detection rates were high (77-100%) for 5/6 of spiked serum. There was a significant association between the qPCR platform and performance. However, certain technical steps and optimal combinations of factors may also impact performance. The good reproducibility and performance demonstrated in this study support the use of Mucorales qPCR as part of the diagnostic strategy for mucormycosis.

Trade-Offs with rehabilitation Effectiveness (REs) and Efficiency (REy) in a sample of Italian disabled persons in a in post-acuity rehabilitation unit.

BACKGROUND: Intensive Rehabilitation Centres, known in Italy as "code 56", admit patients who need to recover from an acute episode. Different Rehabilitation Impact Indices have been proposed as composite rehabilitation outcomes measuring the rate of improvement due to a rehabilitation program. The most widely employed measure the performance of Activities of daily living in rehabilitation is the modified Barthel Index. The Barthel Index-based Rehabilitation Impact Indices are the Rehabilitation Effectiveness and the Rehabilitation Efficiency. AIM: The aim of our study was to evaluate the trade-off between Rehabilitation Effectiveness tayand Rehabilitation Efficiency with respect to the Barthel Index admission score and the Length Of Stay, and their ideal ranges that optimized both indices. METHODS: We retrospectively evaluated data of all patients admitted to intensive rehabilitation unit of the Scientific Institute for Research and Healthcare San Raffaele Pisana of Rome, from January 2006 to March 2018. The primary outcome measures of our study were patient's Rehabilitation Effectiveness and Rehabilitation Efficiency during the hospital stay. RESULTS: A database of 3,466 patients was analysed and the Rehabilitation Effectiveness and Rehabilitation Efficiency indexes were calculated. We calculated the median rank ratio of the Rehabilitation Effectiveness to the Rehabilitation Efficiency against Barthel Index scores. We calculated the median rank ratio of the Rehabilitation Effectiveness to Rehabilitation Efficiency against Barthel Index scores and days of stay. The median rank ratio of the Rehabilitation Effectiveness to the Rehabilitation Efficiency value were 1 in the range of Barthel Index scores from 32 to 42. The median rank ratio of the Rehabilitation Effectiveness to Rehabilitation Efficiency value were 1 for a Length of Stay corresponding to 33 days. CONCLUSIONS: In our study we calculated the Trade-offs between Rehabilitation Effectiveness and Rehabilitation Efficiency with respect to admission Barthel Index Score and Length Of Stay in a population of 3,466 patients affected by orthopedic (1,707) and neurological (1,759) diseases. Every member of the healthcare team should be aware of such trade-offs when they make decisions about rehabilitation services.

Estimating bacteria diversity in different organs of nine species of mosquito by next generation sequencing.

BACKGROUND: Symbiosis in insects is accumulating significant amount of studies: the description of a wide array of mutualistic associations across the evolutionary history of insects suggests that resident microbiota acts as a driving force by affecting several aspects of hosts biology. Among arthropods, mosquito midgut microbiota has been largely investigated, providing crucial insights on the role and implications of host-symbiont relationships. However, limited amount of studies addressed their efforts on the investigation of microbiota colonizing salivary glands and reproductive tracts, crucial organs for pathogen invasion and vertical transmission of symbiotic microorganisms. Using 16S rRNA gene sequencing-based approach, we analysed the microbiota of gut, salivary glands and reproductive tracts of several mosquito species, representing some of the main vectors of diseases, aiming at describing the dynamics of bacterial communities within the individual. RESULTS: We identified a shared core microbiota between different mosquito species, although interesting inter- and intra-species differences were detected. Additionally, our results showed deep divergences between genera, underlining microbiota specificity and adaptation to their host. CONCLUSIONS: The comprehensive landscape of the bacterial microbiota components may ultimately provide crucial insights and novel targets for possible application of symbionts in innovative strategies for the control of vector borne diseases, globally named Symbiotic Control (SC), and suggesting that the holobiont of different mosquito species may significantly vary. Moreover, mosquito species are characterized by distinctive microbiota in different organs, likely reflecting different functions and/or adaptation processes.

Isolation of a Wickerhamomyces anomalus yeast strain from the sandfly Phlebotomus perniciosus, displaying the killer phenotype.

The yeast Wickerhamomyces anomalus has been studied for its wide biotechnological potential, mainly for applications in the food industry. Different strains of W. anomalus have been isolated from diverse habitats and recently from insects, including mosquitoes of medical importance. This paper reports the isolation and phylogenetic characterization of W. anomalus from laboratory-reared adults and larvae of Phlebotomus perniciosus (Diptera: Psychodidae), a main phlebotomine vector of human and canine leishmaniasis. Of 65 yeast strains isolated from P. perniciosus, 15 strains were identified as W. anomalus; one of these was tested for the killer phenotype and demonstrated inhibitory activity against four yeast sensitive strains, as reported for mosquito-isolated strains. The association between P. perniciosus and W. anomalus deserves further investigation in order to explore the possibility that this yeast may exert inhibitory/killing activity against Leishmania spp.

Detection and isolation of the α-proteobacterium Asaia in Culex mosquitoes.

Investigations of microbiota within mosquitoes continue to widen the spectrum of possible symbiont-based applications against vector-borne diseases. In this context, α-proteobacteria of the genus Asaia (Rhodospirillales: Acetobacteraceae) are emerging as possible endosymbiotic candidates, particularly in paratransgenic approaches aimed at interrupting pathogen transmission. Previous studies have shown that Asaia spp. distribution among Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae) mosquitoes displayed positive rates of infection in isolated midguts, salivary glands and reproductive tissues. Similarly, Asaia has been detected in Aedes albopictus (Stegomyia albopicta) and Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) populations. Within the Culex pipiens complex (Diptera: Culicidae), Asaia infection is still largely unexplored. Here, we summarize a preliminary survey of laboratory-reared Cx. pipiens complex and field-collected Culex quinquefasciatus for the presence of Asaia spp., and present the first identification of Asaia in some of the members of the Cx. pipiens complex and the first description in West African populations of Cx. quinquefasciatus.

Efficacy of ultrasound therapy combined with cryotherapy in pain management and rehabilitation in patients with Achilles tendinopathy: a retrospective observational study.

Background: Achilles tendinopathy (AT) is characterized by pain, reduced performance, and swelling in and around the tendon. The aim of our study was to evaluate and compare the effects of ultrasound therapy alone or associated with cryotherapy. Methods: We analyzed retrospectively amateur runner patients who run at least 3 times a week, with medical and ultrasound diagnosis of subacute AT of the midportion. All patients underwent 10 sessions of ultrasounds' therapy with qmd® ultrasound cryo and a therapeutic exercise with stretching and eccentric exercises. The Cryo-Ultrasound Group (CUG, 15, 8M and7/F), during the ultrasound treatment, underwent a session of cryo-ultrasound therapy. The Ultrasound Group (UD, 15, 7M and 8F) only performed ultrasound therapy. Results: All evaluations performed show significant improvement over time in both groups. The CUG shows at T1 a greater increase in pain and function compared to the UG. Friedmann's repeated measures analysis shows that both groups improved when assessed separately over time. From the subsequent post hoc analysis, a statistically significant difference is highlighted between the values evaluated at T0 and T3. Conclusions: The possible simultaneous delivery of the two treatment modalities, in patients suffering from tendinopathies, therefore represents a good possibility of synergistically exploiting their therapeutic actions. Future studies with a larger patient sample and longer follow-up are also needed to better evaluate the benefits of this treatment.

Effectiveness of Kinesiotaping and McConnell taping combined with physical exercise on gait biomechanics in patients with patellofemoral syndrome: non-randomized clinical trial.

Background: Patellofemoral pain syndrome (PFPS) is a pathological condition of the knee, typical of young adults, characterized by diffuse pain in the anterior and / or medial part of the knee. We aimed to examine the effectiveness of the two types of taping in association with therapeutic exercise in relation to the biomechanical parameters, on pain and on functionality of the lower limb in patients with PFPS. Methods: We collected data from patients treated in our outpatient's clinic with two kinds of bandage: the Kinesiotaping group (KG) and the McConnel taping group (MG). All subjects were evaluated trough an optoelectronic system, the Numeric Pain Rating Scale (NPRS), and with the Lower Extremity Functional Scale (LEFS) at baseline before applying the taping (T0), fifteen minutes after applying the bandage (T1), after four weeks of treatment (T2) without applying the bandage and three months after the end of the first treatment period with bandages and exercises (T3). Results: Thirty-five patients (KG 16; MG 19) were included in the study. The most statistically significant changes over time in the LEFS and NPRS values have been recorded in the MG group compared to KG. The average speed and hip rotation showed a statistically significant increase between T3 and T0. Conclusion: The application of the knee bandage for PFPS would appear to show improvement in NPRS and LEFS outcomes in both groups. Furthermore, in this study the MG evidenced better results and significant changes over time than KG.

Do mosquito-associated bacteria of the genus Asaia circulate in humans?

Symbiotic bacteria of the genus Asaia have been proposed as tools for control of mosquito-borne diseases, specifically malaria. However, safety issues are a major concern for paratransgenesis strategies. The aim of this study is to investigate, with immunofluorescence assays and quantitative PCR experiments, whether Asaia spp. is circulating among humans. All human sera and whole blood samples analyzed were negative for Asaia spp., thus suggesting that this organism could be utilized, in the future, as a malaria control tool.

Design, synthesis, and characterization of novel aminoalcohol quinolines with strong in vitro antimalarial activity.

Malaria is the fifth most lethal parasitic infections in the world. Herein, five new series of aminoalcohol quinolines including fifty-two compounds were designed, synthesized and evaluated in vitro against Pf3D7 and PfW2 strains. Among them, fourteen displayed IC50 values below or near of 50.0 nM whatever the strain with selectivity index often superior to 100.17b was found as a promising antimalarial candidate with IC50 values of 14.9 nM and 11.0 nM against respectively Pf3D7 and PfW2 and a selectivity index higher than 770 whatever the cell line is. Further experiments were achieved to confirm the safety and to establish the preliminary ADMET profile of compound 17b before the in vivo study performed on a mouse model of P. berghei ANKA infection. The overall data of this study allowed to establish new structure-activity relationships and the development of novel agents with improved pharmacokinetic properties.

Eucalyptol, an essential oil, reduces contractile activity in rat cardiac muscle.

Eucalyptol is an essential oil that relaxes bronchial and vascular smooth muscle although its direct actions on isolated myocardium have not been reported. We investigated a putative negative inotropic effect of the oil on left ventricular papillary muscles from male Wistar rats weighing 250 to 300 g, as well as its effects on isometric force, rate of force development, time parameters, post-rest potentiation, positive inotropic interventions produced by Ca2+ and isoproterenol, and on tetanic tension. The effects of 0.3 mM eucalyptol on myosin ATPase activity were also investigated. Eucalyptol (0.003 to 0.3 mM) reduced isometric tension, the rate of force development and time parameters. The oil reduced the force developed by steady-state contractions (50% at 0.3 mM) but did not alter sarcoplasmic reticulum function or post-rest contractions and produced a progressive increase in relative potentiation. Increased extracellular Ca2+ concentration (0.62 to 5 mM) and isoproterenol (20 nM) administration counteracted the negative inotropic effects of the oil. The activity of the contractile machinery evaluated by tetanic force development was reduced by 30 to 50% but myosin ATPase activity was not affected by eucalyptol (0.3 mM), supporting the idea of a reduction of sarcolemmal Ca2+ influx. The present results suggest that eucalyptol depresses force development, probably acting as a calcium channel blocker.

Usefulness of (1,3) ß-D-glucan detection in bronchoalveolar lavage samples in Pneumocystis pneumonia and Pneumocystis pulmonary colonization.

OBJECTIVE OF THE STUDY: Recent data demonstrate the usefulness of (1,3) ß-d-glucan (BG) detection in serum samples to distinguish patients developing Pneumocystis pneumonia and patients who are colonized by the fungus. In contrast, data of BG detection in bronchoalveolar lavage (BAL) samples from these patient populations are still rare. PATIENTS: In this context, we determined BG levels in BAL samples from 11 Pneumocystis pneumonia (PCP) patients, 10 colonized patients, and 24 Pneumocystis-uninfected patients. MATERIALS AND METHODS: BG levels were determined on each BAL sample using the Fungitell(®) kit (Associates of Cape Cod, Inc., Cape Cod, MA, USA) according to the manufacturer's instructions applied to serum sample examination. RESULTS: The BG levels in BAL samples from the PCP patient group (mean value 20 588 pg/mL) were significantly higher than those in the colonized patient group (mean value 105 pg/mL) (P=0.0001, Mann-Whitney test) and than those in the Pneumocystis-uninfected patient group (mean value 74 pg/mL) (P<0.0001, Mann-Whitney test). The BG levels in BAL samples from the colonized patient group did not differ significantly from those in the Pneumocystis-uninfected patients group (P=0.21). CONCLUSION: The results suggest that measurements of BAL BG levels may facilitate the differential diagnosis of PCP and pulmonary colonization with Pneumocystis.

Prevalence and causes of hypergastrinemia in primary hyperparathyroidism: a prospective study.

Gastrin levels have been reported to be often increased in patients with primary hyperparathyroidism (PHPT) considered to be caused by hypercalcemia. To determine the prevalence of increased basal gastrin and to investigate its causes, 52 consecutive patients with PHPT were studied prospectively, undergoing a clinical, biochemical, and gastric morphofunctional assessment before any parathyroid surgical procedure. This included evaluation of basal and secretin-stimulated gastrin, basal and pentagastrin-stimulated gastric acid secretion, upper gastrointestinal endoscopy, with histological evaluation for gastritis and Helicobacter pylori infection. Twenty of the 52 PHPT patients (38.5%) had increased fasting gastrin. Further investigation allowed us to clearly demonstrate the causes of hypergastrinemia in 16 of these 20 patients. In 7 of 20 (35%), hypergastrinemia was caused by gastric fundus atrophy; in 3 patients (15%), Zollinger-Ellison syndrome with Multiple Endocrine Neoplasia type I was diagnosed; whereas in another 20% of patients, mild hypergastrinemia was ascribed to Helicobacter pylori gastritis. Finally, in 2 patients, additional clinical history revealed an occasional use of the gastric antisecretory drug omeprazole a few days before the serum gastrin determination. This study shows that the hypercalcemic status per se is not sufficient to produce an increase in fasting gastrin levels. Furthermore, gastric fundus atrophy, and not gastrinoma, is the major cause of relevant (>160 pg/mL) hypergastrinemia.

Biofiltration with buffer-free dialysate.

The presence of acetate in the dialysate appears to be superfluous in the new depurative technique indicated as biofiltration, which consists in a standard hemodialysis with high ultrafiltration combined with the reinfusion of 3-4 1/2 liters of solution containing bicarbonate. The presence of acetate could in fact be contraindicated by a number of potential side effects, metabolic, cardiovascular and biological. Hence, starting from the consideration that in standard biofiltration a buffer is already infused directly as bicarbonate, we tried to overcome the potential hazards of the acetate-containing bath simply by using a dialysate without acetate and by increasing the concentration of bicarbonate in the reinfusate. A cumulative clinical experience of 20 months in 4 patients proved the feasibility and safety of the technique and suggests further advantages over standard biofiltration (better control of acid-base equilibrium, better cardiovascular stability.

Use of a fast EM algorithm for 3D image reconstruction with the YAP-PET tomograph.

OBJECTIVE: We would like to improve the image reconstructions for both signal-to-noise ratio (SNR) and spatial resolution characteristics for the small animal positron emission tomograph YAP-PET, built at the Department of Physics of Ferrara University. The three-dimensional (3D) filtered backprojection (FBP) algorithm, usually used for image reconstruction, has a limited angle restriction due to the tomograph geometry, which causes a serious loss in sensitivity. METHODS: We implemented a 3D iterative reconstruction program using the symmetry and sparse properties of the 'probability matrix', which correlates the emission from each voxel to the detector within a coincidence tube. A fraction only of matrix elements are calculated before the reconstruction and stored on disk: this allows us to avoid on-line computation. A depth dependent function differentiates the voxels in a coincidence tube. Three experimental phantoms with no background were reconstructed by using the program, in comparison with traditionally used FBP. RESULTS: The adopted method allowed us to reduce the computation time significantly. Furthermore, the simple depth dependent function improved the spatial resolution. With 64 x 64 x 20 voxels of 0.625 x 0.625 x 2.0 mm(3) in the field of view, the computation time was less than 4 min per iteration on a Sparc Ultra 450 Workstation, and less than 6 min per iteration on a Mac-PPC G3 300 MHz: the spatial resolution measured with a 0.8 mm diameter 18F-FDG filled capillary reconstructed in this way was 2.0 mm FWHM. By decreasing the voxel size to 0.3125 x 0.3125 x 2.0 mm(3) per voxel the transaxial FWHM was 1.7 mm with a computation time of 15 min per iteration on a Sparc Ultra 450. By using all the acquired data, the SNR improves from 1.3 to 6.0 in the worst measured case, a pair of 0.8mm diameter 18F-FDG filled capillaries, which are 2.5 mm apart each other. CONCLUSION: The adoption of iterative reconstruction allowed us to overcome the loss in sensitivity of previously used FBP: this improved the SNR. The studies of symmetry and sparse properties avoided a severe increase of the reconstruction time and of storing space on disk. This fast EM Algorithm is now routinely used for the image reconstruction with the YAP-PET tomograph.

Localized 100 Hz vibration improves function and reduces upper limb spasticity: a double-blind controlled study.

BACKGROUND: Physical modalities such as vibration has been suggested as possible non-pharmacological way to control spasticity. AIMS: The hypotheses tested were: 1) can a selective vibration of the upper limb flexor antagonist, triceps brachii, reduce the spasticity of the flexor biceps brachii muscle; 2) is its association with physiotherapy better than physiotherapy alone in reducing spasticity and improving function, 3) can this possible effect last for longer than the stimulation period. DESIGN: Randomized double-blind study. SETTING: Rehabilitation Institute, inward patients. POPULATION: Thirty hemiplegic patients affected by upper limb spasticity. METHOD: (VIB + PT) group received physiotherapy plus vibration by means of a pneumatic vibrator applied over the belly of the triceps brachii of the spastic side (contact surface 2 cm2; frequency 100 Hz; amplitude 2 mm; mean pressure 250 mBar). (SHAM + PT) group received physiotherapy and sham vibration. Both groups had 60 minutes of physiotherapy (Kabat techniques) for 5 days a week (from Monday to Friday) for 2 weeks. MAIN OUTCOME MEASURE: Ashworth modified scale for spasticity and robot-aided motor tasks changes for functional modifications were evaluated before starting treatment (T0), 48 hours after the fifth session (T1) and 48 hours after the last session (T2). RESULTS: Fisher's exact test showed a statistically significant greater improvements in the (VIB + PT) group (P=0.0001) compared to in the (SHAM + PT) group after 1 week, as well as after 2 weeks of treatment (P=0.0078) at the Ashworth scale. CONCLUSION: 1) 100 Hz vibration applied to the triceps brachii of a spastic upper limb in association with physiotherapy is able to reduce the spasticity of the flexor agonist, biceps brachii; 2) this association is better than physiotherapy alone in controlling spasticity and improving function; 3) this clinically perceivable reduction of spasticity and function improvement extends (for at least 48 hours) beyond the period of application of the vibration, supporting its possible role in the rehabilitation of spastic hemiplegia. CLINICAL REHABILITATION IMPACT: 100 Hz antagonist muscle vibration, a non-pharmacological treatment, can help physiotherapy to reduce flexors spasticity and improve functions in the rehabilitation of upper limb spasticity.

Effects of eugenol on resting tension of rat atria.

In cardiac and skeletal muscle, eugenol (µM range) blocks excitation-contraction coupling. In skeletal muscle, however, larger doses of eugenol (mM range) induce calcium release from the sarcoplasmic reticulum. The effects of eugenol are therefore dependent on its concentration. In this study, we evaluated the effects of eugenol on the contractility of isolated, quiescent atrial trabeculae from male Wistar rats (250-300 g; n=131) and measured atrial ATP content. Eugenol (1, 3, 5, 7, and 10 mM) increased resting tension in a dose-dependent manner. Ryanodine [100 µM; a specific ryanodine receptor (RyR) blocker] and procaine (30 mM; a nonspecific RyR blocker) did not block the increased resting tension induced by eugenol regardless of whether extracellular calcium was present. The myosin-specific inhibitor 2,3-butanedione monoxime (BDM), however, reversed the increase in resting tension induced by eugenol. In Triton-skinned atrial trabeculae, in which all membranes were solubilized, eugenol did not change resting tension, maximum force produced, or the force vs pCa relationship (pCa=-log [Ca2+]). Given that eugenol reduced ATP concentration, the increase in resting tension observed in this study may have resulted from cooperative activation of cardiac thin filaments by strongly attached cross-bridges (rigor state).

Pain and electrophysiological parameters are improved by combined 830-1064 high-intensity LASER in symptomatic carpal tunnel syndrome versus Transcutaneous Electrical Nerve Stimulation. A randomized controlled study.

AIM: The aim of the study was to compare LASER versus transcutaneous electrical nerve stimulation (TENS) in reducing pain and paraesthesia; and in improving motor and sensory median nerve conduction parameters in mild to moderate carpal tunnel syndrome (CTS). DESIGN: Randomised blinded pilot study. Patients and staff administered treatments and outcome measures were blinded. SETTING: Outpatient; Research and Care Rehabilitation Institute. PARTICIPANTS: Twenty CTS symptomatic patients. INTERVENTIONS: Fifteen sessions of: 1) 100 Hz TENS (30 minutes; rectangular waves; 80 ms width, intensity below muscle contraction); 2) combined 830-1064 nm LASER (radiating dose: 250 J cm-2 delivered to the skin overlying the course of the median nerve at the wrist for 100 s at 25 W (18 W [1064 nm] + 7 W [830 nm]) via a fiber-optic probe with a spot size of ~1 cm2). Outcome measures. Visual analogue scale (VAS) for pain and paresthesia; median nerve distal motor latency and sensory nerve conduction velocity. RESULTS: LASER improved both positive and negative sensory symptoms. TENS induced clinical improvement but this was not statistically significant and was limited to pain reduction. LASER but not TENS favourably modified the neurophysiological parameters. CONCLUSION: High-intensity combined LASER wavelengths of 830 nm and 1064 nm, which produce a better transparency with less scattering and a high energy transfer, are better than TENS in improving both pain and paraesthesia as well as neurophysiological parameters in CTS.

Cell-cell interaction and diversity of emergent behaviours.

Despite myriads of possible gene expression profiles, cells tend to be found in a confined number of expression patterns. The dynamics of Boolean models of gene regulatory networks has proven to be a likely candidate for the description of such self-organisation phenomena. Because cells do not live in isolation, but they constantly shape their functions to adapt to signals from other cells, this raises the question of whether the cooperation among cells entails an expansion or a reduction of their possible steady states. Multi random Boolean networks are introduced here as a model for interaction among cells that might be suitable for the investigation of some generic properties regarding the influence of communication on the diversity of cell behaviours. In spite of its simplicity, the model exhibits a non-obvious phenomenon according to which a moderate exchange of products among adjacent cells fosters the variety of their possible behaviours, which on the other hand are more similar to one another. On the contrary, a more invasive coupling would lead cells towards homogeneity.

Effects of eugenol on resting tension of rat atria

In cardiac and skeletal muscle, eugenol (μM range) blocks excitation-contraction coupling. In skeletal muscle, however, larger doses of eugenol (mM range) induce calcium release from the sarcoplasmic reticulum. The effects of eugenol are therefore dependent on its concentration. In this study, we evaluated the effects of eugenol on the contractility of isolated, quiescent atrial trabeculae from male Wistar rats (250-300 g; n=131) and measured atrial ATP content. Eugenol (1, 3, 5, 7, and 10 mM) increased resting tension in a dose-dependent manner. Ryanodine [100 µM; a specific ryanodine receptor (RyR) blocker] and procaine (30 mM; a nonspecific RyR blocker) did not block the increased resting tension induced by eugenol regardless of whether extracellular calcium was present. The myosin-specific inhibitor 2,3-butanedione monoxime (BDM), however, reversed the increase in resting tension induced by eugenol. In Triton-skinned atrial trabeculae, in which all membranes were solubilized, eugenol did not change resting tension, maximum force produced, or the force vs pCa relationship (pCa=-log [Ca2+]). Given that eugenol reduced ATP concentration, the increase in resting tension observed in this study may have resulted from cooperative activation of cardiac thin filaments by strongly attached cross-bridges (rigor state).