Thermal requirement for development of Sancassania rodionovi (Acari: Acaridae) on mushrooms

The free-living mite species Sancassania rodionovi (Zachvatkin) (Acari: Acaridae), is a serious pest of mushrooms in Iran. Studies were conducted to examine the development of this mite in relation to temperature on two mushroom species: Agaricus bisporus Lange (button mushroom) and Pleurotus ostreatus Kummer (oyster mushroom). The developmental time of this acarid mite was studied at eight constant temperatures, ranging from 5 to 40 degrees C, and developmental rates were modeled as a function of temperature. Sancassania rodionovi completed immature development in 17.35 +/- 0.58 and 20.17 +/- 0.88 d at 25 degrees C on button and oyster mushrooms, respectively. When the mite fed on button mushroom, the rate of development increased gradually from 10 to 35 degrees C. Using a linear model, the developmental zero was estimated to be 3.50 degrees C with a thermal constant of 357.14 degree-days. The Logan 10, Briere 1, and Thermodynamic models adequately described the data for this mite and yielded R2 values >0.95; these models provided estimates of optimum temperature for development of 33.244, 32.145, and 32.148 degrees C, respectively. Understanding the influence of temperature on development of S. rodionovi is discussed with respect to pest management in mushroom production.     

DNA-containing polygonal structures detected in somatic nuclei of Bursaria ovata during preparation to cryptobiosis

Supramolecular chromatin organization of the somatic nucleus (macronucleus--Ma) was studied in a free-living unicellular eukaryotic organism, the ciliate Bursaria ovata Beers 1952, at two late successive stages of its encystment (in the state of preparation to cryptobiosis). A modified Miller's method (Sergejeva et al., 1987) and the same technique in combination with high resolution DNA autoradiography were used. In chromatin spread preparations of Ma, not labeled by 3H-thymidine, numerous electron dense structures (rounded, stick-like and polygonal) were revealed, along with rarely occurring typical supramolecular chromatin structures, such as nucleosomic and not nucleosomic threads, nucleomeres, chromomeres, rosette-like looping chromatin, and electron dense chromonemes (Fig. 1). For DNA visualizing in the revealed polygonal structures, the vegetative cells (trophonts) of B. ovata were inoculated into the culture medium, containing 3H-thymidine and food (ciliates Paramecium caudatum). Here, the ciliates passed through 3-4 successive cell division cycles, thus progressively accumulating the radioactive DNA precursor in Ma. After washing the ciliates in 3H-thymidine-free culture medium, the process of their encystment was induced, and Ma were isolated by hand from the ciliates being at two late successive stages of encystment. Isolated Ma were dispersed in the low ionic solutions, as described elsewhere (Sergejeva et al., 1987). The carbon shadowed electron grids, that contained spread Ma preparations, were individually coated with photographic emulsion, according to the loop interference method (Angelier et al., 1976a; Bouteille, 1976). After a 6 month exposure at 4 degrees C, thymidine incorporation was revealed in fibril crowds, rosette-like structures (Fig. 2), and crystal-like plates of different size and morphology (Fig. 3). In all our experiments, non-specific localization of radioactive DNA precursor was not observed. The above data confirm undoubtedly our earlier assumption (Sergejeva et al., 1987; Sergejeva, Bobyleva, 1988) that the Ma chromatin of Bursaria may undergo crystallization during encystment, i.e. in the state of preparation to cryptobiosis. The present data enable us first to suggest that the looping rosette-like chromatin may be transformed into crystal-like structures ("exotic liquid crystal structures") by means of a peculiar loop packing within the limits of an individual resette (Fig. 2-4), these structural transformations taking place without any visible loop destruction. In this paper, we first describe new morphological types of polygonal plates, differing from those earlier reported elsewhere for the Ma of Bursaria (Sergejeva, Bobylova, 1988), and also from the plates earlier described in studies on liquid crystals both in vivo and in vitro (see: Gianonni et al., 1969; Lerman, 1974; Livolant, 1991: Leforestier et al., 1993, 1997, 1999). The technical approaches used in the present work enabled us to obtain, for the first time, a direct evidence of the presence of DNA in the crystallized structures of somatic nuclei of ciliates during their preparation to cryptobiosis, the DNA-containing polygonal structures being highly morphologically diverse. Further studies into the reasons of this diversity are needed.     

The brain is getting ready for dinner

Every evening, as we get ready for dinner, in addition to the routine behaviors of preparing the meal itself, we also prepare our bodies to cope with the upcoming meal. This could take the form of making restaurant reservations, changing into appropriate attire, washing hands, priming ourselves with an aperitif, or even consciously avoiding snacks as the meal approaches. A study by Johnstone and colleagues in this issue of Cell Metabolism (Johnstone et al., 2006) provides evidence that in parallel to our learned preparatory behaviors, our central nervous system is going through comparable motions as it gets ready for the anticipated meal.     

Unicoloniality in Reticulitermes urbis: a novel feature in a potentially invasive termite species

Social insects are among the world's most successful species at invading of new habitats. A good example of this invasive ability is Reticulitermes (Rhinotermitidae), a prominent group of subterranean termites. As a result of human intervention, i.e. transportation and creation of urban heat islands, Reticulitermes have been able to invade and thrive in cities located in areas where the natural habitat is normally too cold for colonization. They commonly infest man-made structures where they can cause extensive damage.This study was designed to evaluate the invasiveness of Reticulitermes urbis that was probably introduced in France from the Balkans. Invasive potential was assessed on the basis of features typical to invasive social insects, i.e. unicoloniality, low intraspecific aggression, high level of polygyny and colony reproduction by budding. The opportunity to study establishment and spreading processes arose after extensive sampling of an imported Reticulitermes urbis population was performed over the entire city of Domène, France (Rh?ne-Alpes region).For the first time, genetic analysis showed that the termites belonged to a single 'genetic entity' forming a vast colony covering about seven hectares. The colony was structured as an extended family with separate reproductive centres. We speculate that termites were introduced in a single location from which they gradually budded throughout the old town. Based on the absence of aggression among different nests within the colony, we defined this 'genetic entity' as a supercolony.     

Sonic Hedgehog-induced proliferation requires specific Gα inhibitory proteins

Proliferation of cerebellar granular neuronal precursors (CGNPs) is mediated by Sonic Hedgehog (Shh), which activates the Patched and Smoothened (Smo) receptor complex. Although its protein sequence suggests that Smo is a G protein coupled receptor (GPCR), the evidence that this receptor utilizes heterotrimeric G proteins as downstream effectors is controversial. In Drosophila, Gα(i) is required for Hedgehog (Hh) activity, but the involvement of heterotrimeric G proteins in vertebrate Shh signaling has not yet been established. Here, we show that Shh-induced proliferation of rat CGNPs is enhanced strongly by the expression of the active forms of Gα(i/o) proteins (Gα(i1), Gα(i2), Gα(i3), and Gα(o)) but not by members of another class (Gα(12)) of heterotrimeric G proteins. Additionally, the mRNAs of these different Gα(i) members display specific expression patterns in the developing cerebellum; only Gα(i2) and Gα(i3) are substantially expressed in the outer external granular layer, where CGNPs proliferate. Consistent with this, Shh-induced proliferation of CGNPs is reduced significantly by knockdowns of Gα(i2) and Gα(i3) but not by silencing of other members of the Gα(i/o) class. Finally, our results demonstrate that Gα(i2) and Gα(i3) locate to the primary cilium when expressed in CGNP cultures. In summary, we conclude that the proliferative effects of Shh on CGNPs are mediated by the combined activity of Gα(i2) and Gα(i3) proteins.     

Inhibition of calmodulin-dependent cyclic AMP phosphodiesterase by phenoxazines

Phenoxazine derivatives were examined for their ability to inhibit the calmodulin-mediated activation of phosphodiesterase, which is based on the hydrolysis of cAMP to AMP by phosphodiesterase in the presence or absence of inhibitor, followed by quantitative analysis by HPLC method. Anticalmodulin activity of phenoxazines with respect to substitution at C-2 position follows the order: 2-trifluoromethyl>2-chloro>unsubstituted phenoxazines. The interaction of phenoxazines with calmodulin using fluorescence spectroscopy has been performed. Binding study showed that calmodulin has two types of binding sites for phenoxazines. One is high affinity binding site (Kd value 0.07-0.46 microM) and the other, a low affinity binding site (Kd value 0.7-34.5 microM). The change in secondary structure of calmodulin upon binding to phenoxazines was studied by circular dichroism (CD) method, which showed that the percentage of helicity decreased with an extensive change in tertiary structure of calmodulin. Kinetic analysis of the phenoxazine-calmodulin interaction showed that phenoxazines competitively inhibited the activation of phosphodiesterase without affecting Vmax. Thus, these studies showed a good correlation between the ability of phenoxazines to block the activation of phosphodiesterase and their ability to bind to the activator.     

Highly stereoselective synthesis and biological properties of nucleoside analogues bearing a spiro inserted oxirane ring

Starting from 2',5'-di-O-TBDMS-3'-ketouridine 1 or its thymine analogue 2, both xylo (3-10) and ribo (20) epimers of a series of 3"-substituted 3'-spironucleosides have been obtained in good yields and with a total stereoselectivity. Most new compounds were moderately cytotoxic with in some cases slightly selective antiproliferative activities. None of these compounds was active against HIV, but some other antiviral activities against HSV-2, CMV, EBV, or VZV, in the micromolar range, were noted in specific cases.     

Catestatin in rat RVLM is sympathoexcitatory, increases barosensitivity, and attenuates chemosensitivity and the somatosympathetic reflex

The fundamental role and corollary effects of neuropeptides that govern cardiorespiratory control in the brain stem are poorly understood. One such regulatory peptide, catestatin [Cts, human chromogranin A-(352-372)], noncompetitively inhibits nicotinic-cholinergic-stimulated catecholamine release. Previously, we demonstrated the presence of chromogranin A mRNA in brain stem neurons that are important for the maintenance of arterial pressure. In the present study, using immunofluorescence histochemistry, we show that Cts immunoreactivity is colocalized with tyrosine hydroxylase in C1 neurons of the rostral ventrolateral medulla (RVLM, n = 3). Furthermore, we investigated the effects of Cts on resting blood pressure, splanchnic sympathetic nerve activity, phrenic nerve activity, heart rate, and adaptive reflexes. Cts (1 mM in 50 nl or 100 μM in 50-100 nl) was microinjected into the RVLM in urethane-anesthetized, vagotomized, ventilated Sprague-Dawley rats (n = 19). Cardiovascular responses to stimulation of carotid baroreceptors, peripheral chemoreceptors, and the sciatic nerve (somatosympathetic reflex) were analyzed. Cts (1 mM in 50 nl) increased resting arterial pressure (28 ± 3 mmHg at 2 min postinjection), sympathetic nerve activity (15 ± 3% at 2 min postinjection), and phrenic discharge amplitude (31 ± 4% at 10 min postinjection). Cts increased sympathetic barosensitivity 40% (slope increased from -0.05 ± 0.01 before Cts to -0.07 ± 0.01 after Cts) and attenuated the somatosympathetic reflex [1st peak: 36% (from 132 ± 32.1 to 84.0 ± 17.0 μV); 2nd peak: 44% (from 65.1 ± 21.4 to 36.6 ± 14.1 μV)] and chemoreflex (blood pressure response to anoxia decreased 55%, sympathetic response decreased 46%). The results suggest that Cts activates sympathoexcitatory bulbospinal neurons in the RVLM and plays an important regulatory role in adaptive reflexes.