Thioredoxin can influence gene expression by affecting gyrase activity

The expression of many genes of facultatively photosynthetic bacteria of the genus Rhodobacter is controlled by the oxygen tension. Among these are the genes of the puf and puc operons, which encode proteins of the photosynthetic apparatus. Previous results revealed that thioredoxins are involved in the regulated expression of these operons, but it remained unsolved as to the mechanisms by which thioredoxins affect puf and puc expression. Here we show that reduced TrxA of Rhodobacter capsulatus and Rhodobacter sphaeroides and oxidized TrxC of R.capsulatus interact with DNA gyrase and alter its DNA supercoiling activity. While TrxA enhances supercoiling, TrxC exerts a negative effect on this activity. Furthermore, inhibition of gyrase activity strongly reduces puf and puc expression. Our results reveal a new signaling pathway by which oxygen can affect the expression of bacterial genes.     

Dephosphorylation of cofilin is regulated through Ras and requires the combined activities of the Ras-effectors MEK and PI3K

Remodeling of the actin cytoskeleton is crucial for a multitude of cellular functions including cell movement, intracellular transport as well as signal transduction and gene expression processes. Cofilin has been identified as a key mediator of actin reorganization. Its activity is regulated via reversible phosphorylation of ser-3. In a variety of cell types stimulation through particular surface receptors fastly induces the dephosphorylation/activation of cofilin. Yet, the signal transduction cascades linking receptor stimulation with cofilin activation have not been identified so far. Here we show that the GTPase Ras acts as a central regulator of the cofilin dephosphorylation pathway. Thus, stimulation of Ras through platelet-derived growth factor (PDGF) or transient expression of activated Ras-proteins induces the dephosphorylation of cofilin. Importantly, the cooperation of two Ras-initiated signaling pathways is required to induce cofilin dephosphorylation: a Ras-Raf-MAPkinase/Erk-kinase (MEK)- and a Ras-phosphatidylinositol-3-kinase (PI3K)-effector cascade.     

Synthesis, complete characterization, and enantioselective electrokinetic separation of functionalized ruthenium complex enantiomers

Electrokinetic chromatography was employed to separate the enantiomers of two novel functionalized ruthenium(II) complexes with different polypyridyl coordination spheres. The use of anionic carboxymethyl-beta-cyclodextrin as chiral mobile phase additive resulted in maximum efficiency and resolution for the enantiomer separation of both transition metal complexes. The syntheses of the [4-(3-hydroxypropyl)-4'-methyl-2,2'-bipyridine]-bis(2,2'-bipyridine)rethenium(II)-bis(tetrafluoroborate) and [4-(3-hydroxypropyl)-4'-methyl-2,2'-bipyridine]-bis(4,4'-dimethyl-2,2'-bypyridine)ruthenium(II)-bis(tetrafluoroborate) complexes and their complete characterization by means of two-dimensional (1)H and (13)C[(1)H] NMR techniques ((1)H-(1)H COSY and (1)H-(13)C HMQC) as well as elemental analyses and MALDI-TOFMS are described in detail. The functionalized complexes can be used as building blocks for further reactions with polymers, biopolymers, surfaces and nanoparticles.     

Stem cell factor and H2O2 induce GLUT1 translocation in M07e cells

This work aims to elucidate the mechanisms involved in the early activation of glucose transport in hematopoietic M07e cells by stem cell factor (SCF) and a reactive oxygen species (ROS) as H2O2. SCF and H2O2 increase Vmax for glucose transport; this enhancement is due to a higher content in GLUT1 in plasma membranes, possibly through a translocation from intracellular stores. Inhibitors of tyrosine kinases or phospholipase C (PLC) remove glucose transport enhancement and prevent translocation. The inhibitory effect of STI-571 suggests a role for c-kit tyrosine kinase on glucose transport activation not only by SCF, but also by H2O2. On the other hand, neither protein kinase C nor phosphoinositide-3-kinase appear to be involved in the acute activation of glucose transport. Our data suggest that i) in M07e cells, SCF and exogenous H2O2 elicit a short-term activation of glucose transport through a translocation of GLUT1 from intracellular stores to plasma membranes; ii) both stimuli could share at least some signaling pathways leading to glucose uptake activation, involving protein tyrosine kinases and PLC iii) H2O2 could act increasing the level of tyrosine phosphorylation through the inhibition of tyrosine phosphatases and mimicking the regulation role of endogenous ROS.     

Exercise hyperventilation, dyspnea sensation, and ergoreflex activation in lone atrial fibrillation

Lone atrial fibrillation may be associated with daily life disability and exercise limitation. The extracardiac pathophysiology of these effects is poorly explored. In 35 subjects with lone atrial fibrillation (mean age 67 +/- 7 yr), we investigated pulmonary function, symptom-limited cardiopulmonary exercise performance, muscle ergoreflex (handgrip exercise) contribution to ventilation, and brachial artery flow-mediated dilation (as a measure of endothelial function) before and after (average interval 20 +/- 5 days) restoring sinus rhythm with external cardioversion. Respiratory volumes and lung diffusing capacity at rest were within normal limits during both atrial fibrillation and after restoring sinus rhythm. Cardioversion was associated with the following changes: a decrease of the slope of exercise ventilation vs. CO2 production (from 35 +/- 5 to 29 +/- 3; P <0.01) and of dyspnea sensation (Borg score from 4 to 2) and an increase of peak oxygen uptake (Vo2; from 16 +/- 4 to 20 +/- 5 ml.min(-1).kg(-1); P <0.01), Vo2 at anaerobic threshold (from 11 +/- 2 to 13 +/- 2 ml.min(-1).kg(-1); P <0.05), and O2 pulse (from 8 +/- 3 to 11 +/- 3 ml/beat; P <0.01). After cardioversion, the observed improvement in ventilatory efficiency was accompanied by a significant peak end-tidal CO2 increase (from 33 +/- 2 to 37 +/- 2 mmHg; P <0.01) and no changes in dead space-to-tidal volume ratio (from 0.23 +/- 0.03 to 0.23 +/- 0.02; P=not significant). In addition, the ergoreflex contribution to ventilation was remarkably attenuated, and the brachial artery flow-mediated dilatation was significantly augmented (from 0.32 +/- 0.07 to 0.42 +/- 0.08 mm; P <0.01). Ten patients had atrial fibrillation relapse and, compared with values after restoration of regular sinus rhythm, invariably showed worsening of endothelial function, exercise ventilatory efficiency, and muscle ergoreflex contribution to ventilation. In subjects with lone atrial fibrillation, an impairment in ventilatory efficiency appears to be involved in the pathophysiology of exercise limitation, and to be primarily related with a demodulated peripheral control of ventilation.     

Human cystic hydatidosis in Italy: a public health emergency? Past to present

Starting from 1991 the Central Office of Statistics ISTAT, according to changed regulations on notification of infective diseases, stopped reporting official national data on human hydatidosis. On the other hand until then notified data, concerning only about a hundred cases per year in mean during the last decades and just 36 in 1991, appeared unreliable, suggesting a diffusion far from the actual. Owing to specific studies on different research groups it is possible to try to describe a not exhaustive but indicative pattern of trend of human hydatidosis in Italy. More recent publications regarding different regions report an annual mean incidence rate of surgical cases of 9.77/100,000 inhabitants in Sardinia, of 1.57 in Emilia Romagna, of 2.30 in Sicily and of 2.33 in Apulia, the latter encompassing only hepatic cysts treated in the region, irrespective of residence of subjects. A comparison with previous studies regarding the period between 1948-1952 and 1980-84 respectively, indicates an increase in the more recent surveys. However, as even underlined by Authors, former surveys were not all-embracing (for example not including patients from private Clinics etc). On the other hand during time, improvements in surgical as well in anaesthesiological skills have made surgery possible in patients previously inoperable, and particularly in old subjects. Conversely, at present, availability of alternative therapies such as chemotherapy, PAIR or modified PAIR-based techniques may allow cases to avoide surgical surveys. However, actual parasite burden in humans may be estimated by assessing the trend of likely young cysts, as due to recent infections. Rate of pulmonary cysts, that are sure to be young, was of 48.6%/100,000 inhab. in 1952 in Italy, then falling to 18.2 in 1980-84 in the 9 considered regions, to 12.7 in Sicily (1989-91), 6.5 in Emilia-Romagna (1989-93) and 15.7 in Sardinia (1990-95). Another "class" of surely young cysts regards all cysts in young age group that necessarily have been recently infected. Ratio of infection rates in the old/young people was around 1 in 1941-52 but was exceeding 3 in 1980-84. In addition available data from more recent surveys indicate that ratio is at present still relatively low in Sardinia (3.9), where sheep-breeding is widespread, and dog-livestock-man interaction is most likely to exist, and conversely is very high in Emilia-Romagna (5.75) where pastoral economy is not prevalent (just 92,000 sheep). Actual trend both parasitological and in terms of public health would ameliorate if studies were provided with long temporary series. In Sardinia all hydatid cases surgically treated from 1969 to 1995 have been registered and analysed with a standardized methodology. During the considered period incidence appears nearly unvaried (16.9 in 1969 vs 15.8 in 1985), with a fall only for the last 5 years (9.1 in 1995), and with an increase regarding 1984-88 period. Analysis according to cyst localization strongly suggests that fall in the last period is only weakly related to hepatic cysts, that conversely are near completely responsible of rise in 1984-88. On the contrary for pulmonary cysts, i.e. young cysts, a slow but continuous and significant decrease (from 6.5 in 1969 to 1.4 in 1995) is evident. In the same way the above- mentioned old/young ratio rises from 1.7 (1969-79) to 3.9 (1990-95). On the whole analysed data suggest that although parasitic burden, strong in central-south Italy for decades, is now slow lowering due to structural changes and improvements in general conditions of life rather to specific preventive actions, human hydatidosis still represents a public health problem that will continue in the future as, because of latency, incidence rates will stay long relevant.     

Purine nucleosides support the neurite outgrowth of primary rat cerebellar granule cells after hypoxia

Mammalian neurons require a constant supply of oxygen to maintain adequate cellular functions and survival. Following sustained hypoxia during ischemic events in brain, the energy status of neurons and glia is compromised, which may subsequently lead to cell death by apoptosis and necrosis. Concomitant with energy depletion is the formation of the purine nucleoside adenosine, a powerful endogenous neuroprotectant. In this paper the effect of chemical hypoxia on cell survival and neurite outgrowth of primary cerebellar granule cells was investigated. Rotenone, a mitochondrial complex I inhibitor, induced a 30.4 +/- 3.6% loss of viable cells and a 35.0 +/- 4.4% loss of neurite formation of cerebellar granule cells, which was partially restored by the addition of purine nucleosides adenosine, inosine and guanosine. Inosine had the most striking effect of 37.7 +/- 2.9% rescue of viability and 71.2 +/- 18.4% rescue of neurite outgrowth. Data confirm the suggested role of purine nucleosides for the neuronal regeneration of primary brain cells following hypoxic insult.