Downregulation of connexin40 and increased prevalence of atrial arrhythmias in transgenic mice with cardiac-restricted overexpression of tumor necrosis factor

Atrial arrhythmias, primarily atrial fibrillation, have been independently associated with structural remodeling and with inflammation. We hypothesized that sustained inflammatory signaling by tumor necrosis factor (TNF) would lead to alterations both in underlying atrial myocardial structure and in atrial electrical conduction. We performed ECG recording, intracardiac electrophysiology studies, epicardial mapping, and connexin immunohistochemical analyses on transgenic mice with targeted overexpression of TNF in the cardiac compartment (MHCsTNF) and on wild-type (WT) control mice (age 8-16 wk). Atrial and ventricular conduction abnormalities were always evident on ECG in MHCsTNF mice, including a shortened atrioventricular interval with a wide QRS duration secondary to junctional rhythm. Supraventricular arrhythmias were observed in five of eight MHCsTNF mice, whereas none of the mice demonstrated ventricular arrhythmias. No arrhythmias were observed in WT mice. Left ventricular conduction velocity during apical pacing was similar between the two mouse groups. Connexin40 was significantly downregulated in MHCsTNF mice. In contrast, connexin43 density was not significantly altered in MHCsTNF mice, but rather dispersed away from the intercalated disks. In conclusion, sustained inflammatory signaling contributed to atrial structural remodeling and downregulation of connexin40 that was associated with an increased prevalence of atrial arrhythmias.     

Impact of myocardial structure and function postinfarction on diastolic strain measurements: implications for assessment of myocardial viability

Venular control of arteriolar perfusion has been the focus of several investigations in recent years. This study investigated 1) whether endogenous adenosine helps control venule-dependent arteriolar dilation and 2) whether venular leukocyte adherence limits this response via an oxidant-dependent mechanism in which nitric oxide (NO) levels are decreased. Intravital microscopy was used to assess changes in arteriolar diameters and NO levels in rat mesentery. The average resting diameter of arterioles (27.5 +/- 1.0 microm) paired with venules with minimal leukocyte adherence (2.1 +/- 0.3 per 100-microm length) was significantly larger than that of unpaired arterioles (24.5 +/- 0.8 microm) and arterioles (23.3 +/- 1.3 microm) paired with venules with higher leukocyte adherence (9.0 +/- 0.5 per 100-microm length). Local superfusion of adenosine deaminase (ADA) induced significant decreases in diameter and perivascular NO concentration in arterioles closely paired to venules with minimal leukocyte adherence. However, ADA had little effect on arterioles closely paired to venules with high leukocyte adherence or on unpaired arterioles. To determine whether the attenuated response to ADA for the high-adherence group was oxidant dependent, the responses were also observed in arterioles treated with 10(-4) M Tempol. In the high-adherence group, Tempol fully restored NO levels to those of the low-adherence group; however, the ADA-induced constriction remained attenuated, suggesting a possible role for an oxidant-independent vasoconstrictor released from the inflamed venules. These findings suggest that adenosine- and venule-dependent dilation of paired arterioles may be mediated, in part, by NO and inhibited by venular leukocyte adherence.     

Kawal,meat substitute from fermentedCassia obtusifolia leaves

The green leaves ofCassia obtusifolia are fermented to produce a food product, kawal, used by certain tribes of Sudan as a meat substitute. Following fermentation for 2 wk the product is sundried and used when needed. Protein content of kawal is about 20% on dry matter basis. Two microorganisms are most commonly recovered from fermenting as well as dried kawal: the bacteriumBacillus subtilis and a species of the fungusRhizopus. Other uses of the plant are discussed.     

Molecular characterization, technological properties and safety aspects of enterococci from 'Hussuwa', an African fermented sorghum product

AIMS: To identify enterococci from Hussuwa, a Sudanese fermented sorghum product, and determine their technological properties and safety for possible inclusion in a starter culture preparation. METHODS AND RESULTS: Twenty-two Enterococcus isolates from Hussuwa were identified as Enterococcus faecium by using phenotypic and genotypic tests such as 16S rDNA gene sequencing, RAPD-PCR and restriction fragment length polymorphism of the 16S/23S intergenic spacer region fingerprinting. Genotyping revealed that strains were not clonally related and exhibited a considerable degree of genomic diversity. Some strains possessed useful technological properties such as production of bacteriocins and H2O2 or utilization of raffinose and stachyose. None produced alpha-amylase or tannase. A safety investigation revealed that all strains were susceptible to the antibiotics ampicillin, gentamicin, chloramphenicol, tetracycline and streptomycin, but some were resistant to ciprofloxacin, erythromycin, penicillin and vancomycin. Production of biogenic amines or presence of genes encoding virulence determinants occurred in some strains. CONCLUSIONS: Enterococcus faecium strains are associated with fermentation of Sudanese Hussuwa. Some strains exhibited useful technological properties such as production of antimicrobial agents and fermentation of indigestible sugars, which may aid in stabilizing and improving the digestibility of the product respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterococci were shown to play a role in the fermentation of African foods. While beneficial properties of these bacteria are indicated, their presence in this food may also imply a hygienic risk as a result of antimicrobial resistances or presence of virulence determinants.     

Modulation of Calmodulin Plasticity by the Effect of Macromolecular Crowding

In vitro biochemical reactions are most often studied in dilute solution, a poor mimic of the intracellular space of eukaryotic cells, which are crowded with mobile and immobile macromolecules. Such crowded conditions exert volume exclusion and other entropic forces that have the potential to impact chemical equilibria and reaction rates. In this article, we used the well-characterized and ubiquitous molecule calmodulin (CaM) and a combination of theoretical and experimental approaches to address how crowding impacts CaM's conformational plasticity. CaM is a dumbbell-shaped molecule that contains four EF hands (two in the N-lobe and two in the C-lobe) that each could bind Ca²⁺, leading to stabilization of certain substates that favor interactions with other target proteins. Using coarse-grained molecular simulations, we explored the distribution of CaM conformations in the presence of crowding agents. These predictions, in which crowding effects enhance the population of compact structures, were then confirmed in experimental measurements using fluorescence resonance energy transfer techniques of donor- and acceptor-labeled CaM under normal and crowded conditions. Using protein reconstruction methods, we further explored the folding-energy landscape and examined the structural characteristics of CaM at free-energy basins. We discovered that crowding stabilizes several different compact conformations, which reflects the inherent plasticity in CaM's structure. From these results, we suggest that the EF hands in the C-lobe are flexible and can be thought of as a switch, while those in the N-lobe are stiff, analogous to a rheostat. New combinatorial signaling properties may arise from the product of the differential plasticity of the two distinct lobes of CaM in the presence of crowding. We discuss the implications of these results for modulating CaM's ability to bind Ca²⁺ and target proteins.     

Microbiological examination of sebeel water.

Water samples from clay storage jugs ("zeers") located in homes and at public watering stands ("sebeels") at streets, mosques, and schools were examined. Coliforms, fecal coliforms, and fecal streptococci were detected in 100, 69, 88, and 91.56% of the samples, respectively. The general microbiology of the water and some factors affecting microbial load were studied. The predominant bacterial genera of sebeel water were found to be Staphylococcus. Aerococcus, Micrococcus, Streptococcus, Bacillus, Listeria, Lactobacillus, and Arthrobacter. A simple modification of zeer construction was suggested to help improve sanitation.     

Methane fermentation of water hyacinth: effect of retention time and temperature

Summary The influence of retention times of 10, 20, 30 and 40 days on the semi-continuous anaerobic digestion of water hyacinth was tested at the laboratory level. Biogas was successfully produced using 50 and 30 g total solids/l slurries at retention times of 30 and 40 days. The retention time of 40 days proved to be much better than shorter retention times in terms of total gas produced, methane percentage and degree of decomposition of organic matter. The effect of incubation temperatures of 25, 37, 45°C and the variable temperature range of 32 to 42°C on the semicontinuous digestion of water hyacinth was also tested in the laboratory. Biogas could be successfully produced at all temperatures tested except at 45°C. At 37°C and at the variable temperature range of 32 to 42°C gas production was very rapid. Digestion at 25°C was more efficient and gas release was slower. Digestion at 45°C was very erratic.

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Resumen Se estudió, a escala de laboratorio, la influencia de tiempos de retención de 10, 20, 30, y 40 días en la digestión anaerobia semi-continua del jacinto acuático (Eichornia crassipes). A partir de pasta de jacinto acuático con 50 y 30 g de sólidos totales/l de pasta y con tiempos de retención de 30 y 40 días la obtención de gas fue eficaz. La retención de 40 días fue mucho mejor que retenciones mucho más cortas en relación con la producción total de biogás, el porcentaje de metano obtenido y la descomposición de sólidos volátiles. También se estudió el efecto de temperaturas de incubación de 25, 37 y 45°C y de un gradiente de temperaturas de 32–42°C en la fermentación. La producción de biogas fue eficaz a todas las temperaturas ensayadas excepto a 45°C. A 37°C y utilizando el gradiente de temperaturas la obtención de gas fue muy rápida. La digestión a 25°C fue más eficaz pero la liberación de gas más lenta; la fermentación a 45°C dió resultados erráticos.

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Résumé On a testé à l'échelle du laboratoire, l'influence des temps moyens de séjour de 10, 20, 30 et 40 jours sur la biométhanisation semi-continue de la jacinthe d'eau. La production de biogaz était satisfaisante aux concentrations de 50 et 30 g de solides totaux par litre et pour des temps moyens de séjour de 30 et 40 jours. Le temps moyen de séjour de 40 jours s'est révélé bien meilleur que les temps moyens de séjour plus courts en matière de gaz total produit, pourcentage de méthane et conversion de la matière organique. On a également testé au laboratoire l'effet des températures d'incubation de 25, 37 et 45°C et celui d'une température fluctuant entre 32 et 42°C sur la biométhanisation semicontinue de la jacinthe d'eau. La production de biogaz était satisfaisante à toutes les températures sauf celle de 45°C. La production de gaz était très rapide à 37°C et dans la région variable de 32 à 42°C. La biométhanisation à 25°C était plus efficiente mais la vitesse de production du gaz était plus lente. La biométhanisation à 45°C était très erratique.

     

Macromolecular Crowding Modulates Folding Mechanism of α/β Protein Apoflavodoxin

Protein dynamics in cells may be different from those in dilute solutions in vitro, because the environment in cells is highly concentrated with other macromolecules. This volume exclusion because of macromolecular crowding is predicted to affect both equilibrium and kinetic processes involving protein conformational changes. To quantify macromolecular crowding effects on protein folding mechanisms, we investigated the folding energy landscape of an α/β protein, apoflavodoxin, in the presence of inert macromolecular crowding agents, using in silico and in vitro approaches. By means of coarse-grained molecular simulations and topology-based potential interactions, we probed the effects of increased volume fractions of crowding agents (ϕ_c) as well as of crowding agent geometry (sphere or spherocylinder) at high ϕ_c. Parallel kinetic folding experiments with purified Desulfovibro desulfuricans apoflavodoxin in vitro were performed in the presence of Ficoll (sphere) and Dextran (spherocylinder) synthetic crowding agents. In conclusion, we identified the in silico crowding conditions that best enhance protein stability, and discovered that upon manipulation of the crowding conditions, folding routes experiencing topological frustrations can be either enhanced or relieved. Our test-tube experiments confirmed that apoflavodoxin''s time-resolved folding path is modulated by crowding agent geometry. Macromolecular crowding effects may be a tool for the manipulation of protein-folding and function in living cells.     

Integrated multimodal-catheter imaging unveils principal relationships among ventricular electrical activity, anatomy, and function

Multiple imaging modalities are employed independent of one another while managing complex cardiac arrhythmias. To combine electrical, anatomical, and functional imaging in a single catheter system, we developed a balloon catheter that carried 64 electrodes on its surface and an intracardiac echocardiography (ICE) catheter through a central lumen. The catheter system was inserted, and the balloon was inflated inside the left ventricle (LV) of eight dogs with 6-wk-old infarction, created by occlusion in the left anterior descending coronary artery. Anatomy was constructed by ICE imaging (9 MHz) through the balloon. Single-beat noncontact mapping (NCM) was performed via the multielectrode array to reconstruct unipolar endocardial electrograms during sinus rhythm. Standard contact mapping (CM) of the endocardium was also carried out for reference. Myocardial infarction in anterior LV extending from the middle to apical regions was localized both by ICE and NCM and validated by CM and pathology. The overall difference in the activation times between NCM and CM was 3 +/- 1 ms. Unipolar voltage in infarcted middle anterior LV was smaller than the voltage in normal middle inferior LV both by NCM (11 +/- 4 vs. 16 +/- 3 mV; P = 0.002) and CM (11 +/- 3 vs. 20 +/- 4 mV; P < 0.001). Unipolar voltage was also inversely related to infarct transmurality, both by NCM (r = -0.87; P = 0.005) and CM (r = -0.94; P < 0.001). The infarct area by ICE (7.7 +/- 2.9 cm(2)) was in agreement with CM (bipolar voltage, <1 mV; and area, 7.6 +/- 3.3 cm(2); r = 0.80; P = 0.016). Meanwhile, the voltage threshold that depicted the infarct area by NCM was directly related to the smallest unipolar voltage reconstructed within the infarct (r = 0.96; P < 0.001). In conclusion, combining NCM and ICE imaging in a single catheter system is feasible. The preclinical development of such an integrated system and its evaluation in experimental myocardial infarction demonstrate capabilities for single-beat mapping at multiple sites as well as the online assessment of anatomy and myocardial function.