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.     

The in vitro cellular immune response to quartz

It is known that macrophages release into medium in vitro biologically active substances which modulate immune response. In recent years, increase attention has been directed towards the role of prostaglandin in macrophage function. Guinea pig splenic lymphocytes and peritoneal macrophage cultures were incubated with quartz (DQ12), Corundum and aspirin as prostaglandin inhibitor. Lymphocyte proliferation measured by 3H-thymidine incorporation, and lymphokine release by Bendixen-Soborg capillary test (MIF) were applied. EA rosetting and phagocytosis were determined for macrophage function assessment. Quartz suppressed the immune response evidenced by reduced 3H-thymidine incorporation and decreased lymphokine production. Aspirin (as a specific prostaglandin synthesis inhibitor) restored the affected by quartz immunological parameters. This observation gives support to the hypothesis that the immunological response to quartz involves macrophage prostaglandin release, presumably as a first step of lipid peroxidation.     

The effect of oxidants on biomembranes and cellular metabolism

During the reductive process in the tissues, the aerobes generate a number of oxidants. Unless these oxidants are reduced, oxidative damage and cell death would occur. Oxidation of plasma membrane lipids leads to autocatalytic chain reactions which eventually alter the permeability of the cell. The role of oxidative damage in the pathophysiology of diabetic complications and ischemic reperfusion injury of myocardium, especially the changes in the channel activity which may lead to arrhythmia have been studied. Hyperglycemia activates aldose reductase which could efficiently reduce glucose to sorbitol in the presence of NADPH. Since NADPH is also aldose required by glutathione reductase for reducing oxidants, its diversion would lead to membrane lipid oxidation and permeability changes which are probably responsible for diabetic complications such as cataractogenesis, retinopathy, neuropathy etc. Antioxidants such as butylated hydroxy toluene (BHT) and also reductase inhibitors prevent or delay some of these complications. By using patch-clamp technique in isolated frog myocytes, we have shown that hydroxy radicals generated by ferrous sulfate and ascorbate as well as lipid peroxides such as t-butyl hydroperoxide facilitate the entry of Na⁺ by oxidizing Na⁺-channels. Increased intracellular Na⁺ leads to an increase in Na⁺/Ca²⁺ exchange. The increased Na⁺ concentration by itself may produce electrical disturbance which would result in arrhythmia. Increased Ca²⁺ may affect proteases and may help in the conversion of xanthine dehydrogenase to xanthine oxidase, consequently increased production of super oxide radicals. Increased membrane lipid peroxidation and other oxygen free-radical associated membrane damage in myocytes has been demonstrated.     

MEKK2 Kinase Association with 14-3-3 Protein Regulates Activation of c-Jun N-terminal Kinase

MEKK2 (MAP/ERK kinase kinase-2) is a serine/threonine kinase that belongs to the MEKK/STE11 family of MAP kinase kinase kinases (MAP(3)Ks). MEKK2 integrates stress and mitogenic signals to the activation of NF-κB, JNK1/2, p38, and ERK5 pathways. We have found that MEKK2 is regulated through a phosphorylation-dependent association with 14-3-3, a group of adapters that modulate dimerization and association between proteins. We found that MEKK2 was phosphorylated at Thr-283, which resulted in decreased activation loop phosphorylation at Ser-519 and consequently reduced activity. Mechanistically, we found that MEKK2 associated with inactive MEKK2 in the absence of 14-3-3 binding, which led to trans-autophosphorylation of Ser-519. Enforced binding with 14-3-3 reduced Ser-519 trans-autophosphorylation. Expression of T283A MEKK2 within a MEKK2^−/− background enhanced stress-activated c-Jun N-terminal kinase activity while elevating IL-6 expression, but also reduced ERK activation with a corresponding reduced proliferation rate. These results indicate that Thr-283 phosphorylation is an important regulatory mechanism for MEKK2 activation.     

Oregano improves reproductive performance of sows

Natural herbs are being explored as alternatives to antimicrobials. The objective of the present study was to determine the effect of strategic addition of oregano to prefarrowing and lactation diets of sows under field conditions. Alternate farrowing groups were given diets containing 1000 ppm oregano (dried leaf and flower of Origanum vulgare, enriched with 500 g/kg of cold-pressed essential oils of O. vulgare) in prefarrowing and lactation diets. Overall, 801 oregano-treated sows, including 601 primiparous and 1200 multiparous (parity 2.99 +/- 0.43, mean +/- S.E.) and 1809 untreated control sows (705 primiparous and 1104 multiparous; parity 3.04 +/- 0.38), were used. Sows fed oregano had lower annual sow mortality rate (4.02 +/- 0.4% versus 6.92 +/- 1.11%, mean +/- S.E.; P = 0.003), lower sow culling rate during lactation (8.01 +/- 1.11% versus 14.02 +/- 1.33%, P = 0.02), increased farrowing rate (77.02 +/- 2.31% versus 69.91 +/- 2.32%, P = 0.01), increased number of liveborn piglets per litter (10.49 +/- 1.5 versus 9.95 +/- 1.22, P < 0.05), and decreased stillbirth rate (0.909 +/- 0.01 versus 0.807 +/- 0.01, P = 0.05). In addition, multiparous sows fed oregano had higher (P = 0.04) daily voluntary feed intake compared to non-treated sows (7.7 +/- 0.32 kg versus 7.0 +/- 0.42 kg, P = 0.04). Additional studies are needed to elucidate the effects of oregano on the gastrointestinal, immune and urogenital system in swine and to determine if it has any adverse effects.     

Condensation of interphase chromatin in nuclei of synchronized chinese hamster ovary (CHO-K1) cells

Reversibly permeabilized cells have been used to visualize interphase chromatin structures in the presence and absence of biotinylated nucleotides. By reversing permeabilization, it was possible to confirm the existence of a flexible chromatin folding pattern through a series of transient geometric forms such as supercoiled, circular forms, chromatin bodies, thin and thick fibers, and elongated chromosomes. Our results show that the incorporation of biotin-11-dUTP interferes with chromatin condensation, leading to the accumulation of decondensed chromatin structures. Chromatin condensation without nucleotide incorporation was also studied in cell populations synchronized by centrifugal elutriation. After reversal of permeabilization, nuclei were isolated and chromatin structures were visualized after DAPI staining by fluorescent microscopy. Decondensed veil-like structures were observed in the early S phase (at an average C-value of 2.21), supercoiled chromatin later in the early S (2, 55 C), fibrous structures in the early mid S phase (2, 76 C), ribboned structures in the mid-S phase (2, 98 C), continuous chromatin strings later in the mid-S phase (3,28), elongated prechromosomes in the late S-phase (3, 72 C), precondensed chromosomes at the end and after the S phase (3, 99 C). Fluorescent microscopy revealed that neither interphase nor metaphase chromosomes are separate entities but form a linear array arranged in a semicircle. Linear arrangement was confirmed by computer image analysis.