Organic acid and probiotic derived from grass silage improved egg quality in Pegagan laying duck: a research note.

The aim of this study was to evaluate the effect of supplementation of organic acid and probiotic derived from grass silage on the egg quality of duck. Seventy-two Pegagan laying ducks (average age: 24 weeks) were randomly allocated to six treatment groups: basal diet, basal diet + organic acid, basal diet + probiotic, basal diet + tetracycline, basal diet + probiotic + organic acid, and basal diet + organic acid + tetracycline. The result showed that the feeding diets containing probiotics and organic acid significantly (P < 0.05) reduced yolk fat and yolk cholesterol and increased eggshell weight, egg index, yolk color score, Haugh unit, and protein content. However, egg weight, albumen weight, yolk weight, albumen index, yolk index, and water content were not significantly (P > 0.05) different. It is concluded that dietary supplementation of organic acid and probiotic derived from grass silage improved egg quality in terms of yolk color score, fat, and cholesterol content.

A Computational Study of the Electrophysiological Substrate in Patients Suffering From Atrial Fibrillation.

In the context of cardiac electrophysiology, we propose a novel computational approach to highlight and explain the long-debated mechanisms behind atrial fibrillation (AF) and to reliably numerically predict its induction and sustainment. A key role is played, in this respect, by a new way of setting a parametrization of electrophysiological mathematical models based on conduction velocities; these latter are estimated from high-density mapping data, which provide a detailed characterization of patients' electrophysiological substrate during sinus rhythm. We integrate numerically approximated conduction velocities into a mathematical model consisting of a coupled system of partial and ordinary differential equations, formed by the monodomain equation and the Courtemanche-Ramirez-Nattel model. Our new model parametrization is then adopted to predict the formation and self-sustainment of localized reentries characterizing atrial fibrillation, by numerically simulating the onset of ectopic beats from the pulmonary veins. We investigate the paroxysmal and the persistent form of AF starting from electro-anatomical maps of two patients. The model's response to stimulation shows how substrate characteristics play a key role in inducing and sustaining these arrhythmias. Localized reentries are less frequent and less stable in case of paroxysmal AF, while they tend to anchor themselves in areas affected by severe slow conduction in case of persistent AF.

Simultaneous endo-epicardial-high density mapping of ventricular tachycardia with the use of multi-electrode mapping catheters.

We herein describe a case of VT in an ischemic cardiomyopathy patient utilizing a novel technique of simultaneous endo-epicardial high-density mapping. Epicardial access, in addition to retrograde aortic endocardial access, was performed. Following the creation of the respective endocardial and epicardial electrical substrate maps, activation mapping of the VT was then performed simultaneously with two overlapping HD Grid catheters. Full diastolic pathway mapping was achieved, with EGMs recorded in all segments of the diastolic interval, on the epicardial surface. This case highlights the feasibility of a technique that can reliably detect the presence of activation bridges in a 3D model of VT.

European multicentre experience of staged hybrid atrial fibrillation ablation for the treatment of persistent and longstanding persistent atrial fibrillation.

The management of non-paroxysmal atrial fibrillation (AF) remains controversial. We examined the efficacy and safety of the 2 stage Hybrid AF ablation approach by analysing the largest series of this technique reported so far. METHODS: The approach aims to electrically isolate the left atrial posterior wall incorporating the pulmonary veins ('box-set'pattern). An initial video-assisted thoracoscopic (VATS) epicardial ablation is followed after a minimum of 8 weeks by endocardial radiofrequency catheter ablation. RESULTS: Of 175 patients from 4 European cardiothoracic centers, who underwent the surgical (COBRA Fusion, AtriCure Inc) 1st stage ablation, 166 went on to complete 2nd stage catheter ablation. At median follow up of 18 months post 2nd stage procedure 93/166 (56%) had remained free of AF or atrial tachycardia (AT) recurrence off antiarrhythmic drugs. 110/175 62.9% were in sinus rhythm off all antiarrhythmic drugs at last clinic follow-up (132/175 75.4% including those on antiarrhythmic drugs). 18 patients (10.8%) underwent a further re-do ablation (mean of 1.1 ablations per patient) 105/166 (63%) remained free of AF/AT recurrence off antiarrhythmic drugs following last ablation procedure.Latterly, ILRs have been implanted in patients (n = 56); 60% have remained fully arrhythmia free and 80% have shown AF burden < 5% at a median 14 months follow-up [IQR: 13.5 (8-21.5)]. Only 10.9% have reverted to persistent AF. 5 patients (2.9%) had a perioperative stroke and 4 patients (2.3%) exhibited persistent weakness of the right hemidiaphragm following stage 1 VATS epicardial ablation. One patient died following stroke (overall mortality 0.6%). CONCLUSIONS: In patients with non-paroxysmal AF with unfavourable characteristics for catheter ablation, the staged hybrid approach results in acceptable levels of freedom from recurrent atrial arrhythmia, however, complication rates are higher than with catheter ablation alone.

Updated clinical overview on cardiac laminopathies: an electrical and mechanical disease.

Cardiac laminopathies, associated with mutations in the LMNA gene, encompass a wide spectrum of clinical manifestations, involving electrical and mechanical alterations of cardiomyocytes. Thus, dilated cardiomyopathy, bradyarrhythmias and atrial or ventricular tachyarrhythmias may occur in a number of combined phenotypes. Nowadays, some attempt has been made to identify clinical predictors for the most life-threatening complications of LMNA-associated heart disease, i.e. sudden cardiac death and end-stage heart failure. The goal of this manuscript is to combine the most recent evidences in an updated review to show the state-of-the-art of such a complex disease group. This is supposed to be the starting point to collect more data and design new ad hoc studies to identify clinically useful predictors to stratify risk in mutation carriers, including probands and their asymptomatic relatives.

The size of DNA molecules and chromatin organization in the macronucleus of the ciliate Didinium nasutum (Ciliophora).

Pulsed-field gel electrophoresis (PFGE) was applied to analyze the molecular karyotype of the ciliate Didinium nasutum. The data obtained indicate that D. nasutum belongs to the ciliate species with subchromosomal macronuclear genome organization. No short "gene-sized" DNA molecules were detected. Macronuclear DNAs formed a continuous spectrum from 50 kbp to approximately 1,000 kbp in size with a peak plateau between 250 and 400 kbp. The macronuclear DNA molecules were packed into chromatin bodies of 80-265 nm in size. Comparison of the PFGE and electron microscopic data shows that most if not all chromatin bodies contain more than one DNA molecule.

Draft Genome Sequence of Pseudomonas mediterranea Strain CFBP 5447T, a Producer of Filmable Medium-Chain-Length Polyhydroxyalkanoates.

Pseudomonas mediterranea strain CFBP 5447(T) is a phytopathogenic bacterium isolated from tomato plants affected by pith necrosis disease. Moreover, its ability to produce medium-chain-length polyhydroxyalkanoates (mcl-PHAs) in culture from different carbon sources and valuable microbial products, such as cyclic lipopeptides, has been well documented. Here, we report the first draft genome sequence of this species.

Draft genome sequence of Pseudomonas corrugata, a phytopathogenic bacterium with potential industrial applications.

Pseudomonas corrugata was first described as the causal agent of a tomato disease called 'pith necrosis' yet it is considered as a biological resource in various fields such as biocontrol of plant diseases and production of industrially promising microbial biopolymers (mcl-PHA). Here we report the first draft genome sequence of this species.

Rhythm-symptom correlation in patients on continuous monitoring after catheter ablation of atrial fibrillation.

INTRODUCTION: Correlation between symptoms and atrial fibrillation (AF) episodes after catheter ablation may have clinical relevance, especially for anticoagulation usage. The aim of our project was to analyze the relationship between symptoms and AF recurrences in unselected patients following AF catheter ablation during long-term follow-up. METHODS AND RESULTS: One hundred and forty-three consecutive patients (mean age 59 ± 9 years, 85% male) were implanted with a continuous cardiac monitor (RevealXT, Medtronic Inc., Minneapolis, MN, USA) following first pulmonary vein ablation procedure. Device data were downloaded every 3 months and correlated to patients' symptom diary. AF was paroxysmal in 55% and persistent in 45%. At a mean follow-up of 14 ± 6 months, 98/143 (69%) patients had at least one AF recurrence. Among these, 53 (54%) reported AF-related symptoms while 45 (46%) were totally asymptomatic. Conversely, 13 (29%) out of 45 patients without AF recurrences reported symptoms. Globally, a significant reduction of symptoms (from 82% at baseline to 44% at last follow-up; P < 0.0001) was observed. SF-12 questionnaire showed a significant improvement of physical and mental functioning (respectively 44.5 ± 8.5 vs 51.0 ± 6.7, and 45.7 ± 9.3 vs 49.2 ± 6.1, P < 0.05 baseline vs last follow-up). At the 12-month follow-up, 80% and 77% of patients were on AAD and anticoagulant drugs, respectively. There were not differences in AAD usage in symptomatic and asymptomatic patients. CONCLUSIONS: Continuous ECG monitoring is a valuable tool for long-term follow-up after AF catheter ablation facilitating reliable assessment of symptomatic and asymptomatic AF episodes. This may have clinical implications with regards to anticoagulation therapy in high-risk patients.

Quantitative analysis of nucleolar chromatin distribution in the complex convoluted nucleoli of Didinium nasutum (Ciliophora).

We have earlier shown that the typical Didinium nasutum nucleolus is a complex convoluted branched domain, comprising a dense fibrillar component located at the periphery of the nucleolus and a granular component located in the central part. Here our main interest was to study quantitatively the spatial distribution of nucleolar chromatin structures in these convoluted nucleoli. There are no "classical" fibrillar centers in D.nasutum nucleoli. The spatial distribution of nucleolar chromatin bodies, which play the role of nucleolar organizers in the macronucleus of D.nasutum, was studied using 3D reconstructions based on serial ultrathin sections. The relative number of nucleolar chromatin bodies was determined in macronuclei of recently fed, starved D.nasutum cells and in resting cysts. This parameter is shown to correlate with the activity of the nucleolus. However, the relative number of nucleolar chromatin bodies in different regions of the same convoluted nucleolus is approximately the same. This finding suggests equal activity in different parts of the nucleolar domain and indicates the existence of some molecular mechanism enabling it to synchronize this activity in D. nasutum nucleoli. Our data show that D. nasutum nucleoli display bipartite structure. All nucleolar chromatin bodies are shown to be located outside of nucleoli, at the periphery of the fibrillar component.

Quantitative analysis of nucleolar chromatin distribution in the complex convoluted nucleoli of Didinium nasutum (Ciliophora)

We have earlier shown that the typical Didinium nasutum nucleolus is a complex convoluted branched domain, comprising a dense fibrillar component located at the periphery of the nucleolus and a granular component located in the central part. Here our main interest was to study quantitatively the spatial distribution of nucleolar chromatin structures in these convoluted nucleoli. There are no "classical" fibrillar centers in D.nasutum nucleoli. The spatial distribution of nucleolar chromatin bodies, which play the role of nucleolar organizers in the macronucleus of D.nasutum, was studied using 3D reconstructions based on serial ultrathin sections. The relative number of nucleolar chromatin bodies was determined in macronuclei of recently fed, starved D.nasutum cells and in resting cysts. This parameter is shown to correlate with the activity of the nucleolus. However, the relative number of nucleolar chromatin bodies in different regions of the same convoluted nucleolus is approximately the same. This finding suggests equal activity in different parts of the nucleolar domain and indicates the existence of some molecular mechanism enabling it to synchronize this activity in D. nasutum nucleoli. Our data show that D. nasutum nucleoli display bipartite structure. All nucleolar chromatin bodies are shown to be located outside of nucleoli, at the periphery of the fibrillar component.

Role of calcitonin gene-related peptide and substance P in different models of pain.

Calcitonin gene-related peptide (CGRP) and substance P (SP) play an important role in the development of pain and hyperalgesia. Experimental models have demonstrated that nitroglycerin (NTG)--a nitric oxide donor--provokes a hyperalgesic state, probably via the activation of second-order neurons in the nucleus trigeminalis caudalis. In order to gain further insight into the role of CGRP and SP in different types of experimental pain, we evaluated and compared changes in immunoreactivity (-ir) for these two neuropeptides at different levels of the central nervous system [nucleus trigeminalis caudalis (NTC) and dorsal horns of the lumbar spinal cord] in two animal models of hyperalgesia: systemic NTG administration and formalin test. Following NTG administration, CGRP-ir decreased steadily in the NTC, whereas SP-ir increased transiently. In the lumbar dorsal horns, NTG induced a decrease in SP-ir 1 h after its administration. Formalin injection induced an ipsilateral increase in both CGRP and SP immunostaining at 1 and 2 h in the lumbar dorsal horns. In the NTC, a significant decrease in CGRP-ir was observed at 1 h. The changes in the staining intensities were paralleled by changes in the numbers of CGRP and of SP varicosities in both the NTC and the lumbar dorsal horns. These findings show specific changes in CGRP and SP at different levels of the central nervous system in the different models of pain. In the case of the formalin test, the changes involve both neuropeptides synchronously and to the same extent, whereas in the case of NTG administration, CGRP seems to play a more prevalent and long-lasting role, particularly at the NTC level.

First Report of Leaf Spot and Blight Caused by Ralstonia pickettii on Bird of Paradise Tree in Italy.

Bird of Paradise tree (Strelitzia alba (L. f.) Skeels) is an ornamental perennial tropical plant grown in southern Italy. In the summer of 2006 and 2007, a widespread, severe leaf disease was observed on seedlings and 1- to 2-year-old plants in two glasshouses located in eastern Sicily. Disease incidence ranged from 10 to 25%. Symptoms on the leaves consisted of dark brown-to-black stripes of varying length and found between the lateral veins. Lesions sometimes coalesced into a large area of necrotic tissue. Symptomatic tissues were ground in a drop of sterile distilled water (SDW) with a scalpel. Suspensions were streaked on King's medium B (KB), nutrient agar, and yeast extract nutrient agar (2). Isolated strains were gram negative and oxidase positive, non-levan, negative in tobacco hypersensitivity test, white and nonmucoid on yeast dextrose calcium carbonate agar, did not produce fluorescent pigments on KB, and utilized glucose, mannitol, trehalose, arabinose, mannose, and N-acetylglucosamine. Bacterial strains were identified as Ralstonia pickettii by using the Biolog Identification System (MicroLogTM System Release 4.2; Biolog, Inc., Hayward, CA) with a similarity index ranging from 0.52 to 0.67. For an additional confirmation of identity, the small subunit rRNA gene (SSUrDNA) was amplified with primers 530F and Uni 1492R (1). The resulting nucleotide sequence was compared with sequences deposited in GenBank and showed the highest identity (99%) to sequences of R. pickettii strains. Pathogenicity tests were performed on 20 cm tall potted plants. Four S. alba plants were inoculated by infiltrating leaf veins with bacterial suspensions for each of the four isolates (107 CFU ml-1 in SDW) with a 25-gauge needle and syringe. Plants were placed in polyethylene bags 1 day before inoculation and maintained there for 3 days after inoculation. Four control plants were inoculated with SDW. Water-soaked areas in the lateral veins of leaves were observed in all inoculated plants 4 days after inoculation. Within 10 days, dark brown-to-black stripes that coalesced into dark necrotic areas were observed. All isolates induced similar symptoms. Control plants did not show any symptoms. The pathogen was reisolated from symptomatic tissue and identified as R. pickettii by Biolog. A similar disease on S. reginae caused by a Pseudomonas sp. was previously reported from Florida (3). To our knowledge, this is the first record in the world of leaf spot and blight caused by R. pickettii. References: (1) D. J. Lane. 16S/23S rRNA sequencing. Page 115 in: Nucleic Acid Techniques in Bacterial Systematics. E. Stackebrandt and M. Goodfellow, eds. John Wiley and Sons, NY, 1991. (2) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. The American Phytopathological Society, St. Paul, MN, 2001. (3) C. Wehlburg. Plant Dis. Rep. 55:447, 1971.

Combination of dexamethasone and etanercept reduces secondary damage in experimental spinal cord trauma.

The aim of our study was to evaluate the therapeutic efficacy of combination therapy with etanercept and dexamethasone (DEX) in vivo in experimental murine model of spinal cord trauma, which was induced by the application of vascular clips (force of 24 g) to the dura via a four-level T5-T8 laminectomy. Spinal cord injury in mice resulted in severe trauma characterized by edema, neutrophil infiltration, and cytokine production followed by recruitment of other inflammatory cells, production of inflammation mediators, tissue damage, apoptosis and disease. Treatment of the mice with etanercept (1.25 mg/kg) and DEX (0.025 mg/kg) when administered as a combination therapy but not as a single treatment significantly reduced the degree of (1) spinal cord inflammation and tissue injury (histological score), (2) infiltration of neutrophils (MPO evaluation), (3) inducible nitric oxide synthase, nitrotyrosine, and cytokines expression (tumor necrosis factor-alpha and interleukin-1 beta), (4) and apoptosis (Terminal deoxynucleotidyltransferase-mediated UTP end labeling staining, Fas-ligand expression and Bax and Bcl-2 expression). In a separate set of experiments we have also clearly demonstrated that the combination therapy significantly ameliorated the recovery of limb function (evaluated by motor recovery score). Taken together, our results clearly demonstrate for the first time that strategies targeting multiple proinflammatory pathways may be more effective than a single effector molecule for the treatment of spinal cord trauma.

Intrathecal baclofen therapy in patients with severe spasticity.

Spasticity has been described as "a motor disorder, characterized by a velocity-dependent increase in tonic stretch reflexes (muscle tone) with exaggerated tendon jerks, resulting from hyper-excitability of the stretch reflex as one component of the upper motor neuron syndrome". In patients with complete spinal cord lesions, severe untreatable spasticity can make movement, sitting and hygiene difficult or impossible while it may alter gait and personal care in patients with partial lesions. From a clinical point of view, it is useful to distinguish spinal cord spasticity from supraspinal spasticity. Traditionally, the Ashworth scale is the most widely used to quantify the tone of single muscles. In order to quantify hypereflexia, the Reflex Scale is also used. In the spinal spasticity which is characterized by spasms, the Spasm Frequency Scale is useful in order to monitor their frequency. Initially, management of spasticity is based on non-invasive treatments that later become more invasive. The first approach. the conservative treatment, usually includes elimination of the nociceptive stimuli, rehabilitative therapy (physical and occupational), orthopaedic prostheses and plaster corsets. These treatments, do not resolve spasticity in about 33% of cases. In these severe cases, more invasive procedures such as muscle infiltrations with botulin toxin and intrathecal baclofen infusion can be used.

Infection of algae-free Climacostomum virens with symbiotic Chlorella sp. isolated from algae-containing C. virens.

The ciliate Climacostomum virens normally contains algae as symbionts in its cytoplasm and retains them over many generations. An aposymbiotic strain of C. virens which cannot re-establish a new symbiotic association by ingestion of algae derived from green Climacostomum was recently isolated in our laboratory. Results of infection experiments showed that all newly ingested, potentially symbiotic algae were digested in food vacuoles. To clarify whether these ciliates have completely lost their ability to sustain symbiosis with algae or whether this ability can eventually be re-established, infection experiments were performed using a microinjection technique. We have achieved successful infection of algae-free Climacostomum using this method. The endosymbiont population was established in ciliates from as few as 3-5 injected algae, which have retained an intact perialgal vacuole membrane around them. To our knowledge, this is the first evidence of successful infection of aposymbiotic ciliates with algae by microinjection.

Asymmetric Guillain-Barré syndrome.

Guillain-Barré syndrome (GBS) is a heterogeneous disorder according to clinical, electrophysiological, immunologic and pathologic findings. It has usually been considered as an immune-mediated polyneuropathy clinically characterised by acute symmetric muscle weakness and areflexia. We describe a patient who, after a Campylobacter jejuni infection, developed an acute motor-sensory neuropathy with marked and persistent asymmetry of clinical and electrophysiological findings. He had a high titre of anti-GM1 IgG antibodies and cytoalbuminologic dissociation and was responsive to intravenous immunoglobulins. Investigations and three years of follow-up excluded mimics of GBS. Tendon areflexia has recently been challenged as a mandatory diagnostic criterion in GBS; likewise marked and persistent motor asymmetry does not exclude the diagnosis of GBS.

Nucleolar apparatus in the macronucleus of Didinium nasutum (Ciliata): EM and 3D reconstruction.

The three-dimensional (3D) organization of nucleoli in the somatic nuclei (macronuclei) of recently fed and starved Didinium nasutum was reconstructed on the basis of serial ultra-thin sections. It was shown that nucleoli, looking on the single sections like individual separate structures, appeared to be parts of the large complicated branchy nucleolar networks. A 30 h starvation did not lead to disintegration of this network, but stimulated formation of numerous vacuoles in the granular component of nucleoli, which becomes more condensed. Unlike starved D. nasutum, in fed ciliates numerous holes appeared in the fibrillar component located at the periphery of nucleoli. These holes may presumably serve as channels for transporting newly synthesized rRNA. To our knowledge, this is the first report of a 3D reconstruction of the nucleolar apparatus in ciliates.

Identification and Detection of Phoma tracheiphila, Causal Agent of Citrus Mal Secco Disease, by Real-Time Polymerase Chain Reaction.

Phoma tracheiphila is the causal agent of a tracheomycotic disease of citrus called mal secco causing the dieback of twigs and branches. This pathogen is of quarantine concern; therefore, fast and reliable protocols are required to detect it promptly. A specific primer pair and a dual-labeled fluorogenic probe were used in a real-time polymerase chain reaction (PCR) with the Cepheid Smart Cycler II System (Transportable Device TD configuration) to detect this fungus in citrus samples. Real-time PCR assay was compared to modified conventional PCR assay. The sensitivity of the former was evaluated by testing P. tracheiphila DNA dilutions, and the minimum amount detectable was about 500 fg, whereas the linear quantification range was within 100 ng to 1 pg. Conventional PCR sensitivity was 10 pg. Conventional and real-time PCR successfully detected the fungus in woody samples of naturally infected lemon and artificially inoculated sour orange seedlings. Nevertheless, real-time PCR was about 10- to 20-fold more sensitive than conventional PCR, and preliminary results indicate that the former technique achieves quantitative monitoring of the fungus in tissues. Simple and rapid procedures to obtain suitable DNA samples from fungal cultures and citrus woody samples for PCR assays enable diagnosis to be completed in a short time.

Non-contact left ventricular endocardial mapping for cardiac resynchronisation therapy: a "slow conduction" towards the fast solution.

Cardiac resynchronisation therapy can help to improve left ventricular function in patients with heart failure, but only if those regions of myocardium which are mostly compromised by electromechanical desynchronisation can be identified and effectively stimulated.