Glutamatergic chemical transmission: look! Here, there, and anywhere

Vesicular glutamate transporter (VGLUT) is responsible for the active transport of L-glutamate in synaptic vesicles and thus plays an essential role in the glutamatergic chemical transmission in the central nervous system. VGLUT comprises three isoforms, VGLUT1, 2, and 3, and is a potential marker for the glutamatergic phenotype. Recent studies indicated that VGLUT is also expressed in non-neuronal cells, and localized with various organelles such as synaptic-like microvesicles in the pineal gland, and hormone-containing secretory granules in endocrine cells. L-Glutamate is stored in these organelles, secreted upon various forms of stimulation, and then acts as a paracrine-like modulator. Thus, VGLUTs highlight a novel framework of glutamatergic signaling and reveal its diverse modes of action.     

Caenorhabditis elegans DNA does not contain 5-methylcytosine at any time during development or aging.

DNA, isolated from age-synchronous senescent populations of Caenorhabditis elegans has been quantitatively and qualitatively analyzed for the presence of 5-methylcytosine. High performance liquid chromatography on two wild-type and several mutant strains of C. elegans failed to detect any 5-methylcytosine. The restriction endonuclease isoschizomers, HpaII and MspI, were used to digest genomic DNA after CsCl purification and failed to detect any 5' cytosine methylation at any age. We conclude that C. elegans does not contain detectable (0.01 mole percent) levels of 5-methylcytosine.     

Microclimatic indexes and limits of safe exposure in work activity at risk of thermal stress

The working time in work places thermically uncomfortable must be limited in order to prevent the risk of thermal stress. In a previous study we have demonstrated that the best method for calculation of the safe exposure times is based on the required sweat rate index. In the present work we verified in different work places termically uncomfortable, grouped for equivalent values of WBGT index, the variability of the safe exposure times calculated by means of a method just now mentioned for various levels of energetic expenditure. The results point out the possibility of a mathematical relation between energetic expenditure, WBGT index and mean of safe exposure times.     

Introducing plants for bee-keeping at any cost? – Assessment of Phacelia tanacetifolia as nectar source plant under xeric Mediterranean conditions

 We assessed nectar and honey potential of Phacelia tanacetifolia, an American plant, under Mediterranean conditions. Its flowering occurred during the major flowering season unless intensely irrigated, whereas duration and flower life span were shorter compared to continental conditions. Mediterranean climate limitations to nectar secretion were assessed on plants grown under natural conditions (xeric) vs. regular irrigation. May flowering xeric plants produced less nectar per flower than the irrigated ones, but had the same nectar potential per surface area. On the contrary, at the cost of intense irrigation, July flowering plants produced much higher nectar per flower and per surface area compared to xeric ones. In all flowering periods and sections honeybees were the most constant and numerous visitors, visiting the flowers mainly for nectar, whereas solitary bees were scarce. Based on our results, we suggest that although the plant may be a good nectar source for honeybees in some cases, we have serious reservations for a generalised use in the Mediterranean. Received September 3, 2002; accepted November 18, 2002 Published online: June 2, 2003     

The influence of conditions at the time of mating on reproduction of commercial pigs

The objective of this study was to compare a new mating system, called the Detection-Mating Area (DM Area), and a conventional mating system on the long-term reproductive performance of commercial pigs. The DM Area treatment basically involved detecting oestrous females in an arena closely surrounded on two sides by boars and mating these females in this arena. This mating system was designed to improve the physical and sexual environments of the pigs at mating. In contrast, the conventional treatment involved conducting oestrus detection and mating in the boar's accommodation pen. The study was conducted over an 18-month period at a commercial farm that housed 2400 breeding female pigs.

In order to control for the effects of the stockperson, an analysis was conducted on the reproductive performance of female pigs in which one stockperson assisted the matings in both treatments over a 12-month period. Gilts mated in the DM Area treatment had a higher (P<0.05) total litter size and a higher (P<0.01) litter size born alive than gilts mated in the conventional treatment (10.31 vs. 8.96 and 9.50 vs. 8.29, respectively). Although gilts in the DM Area treatment had a higher farrowing rate (93.2 vs. 87.9%), this difference was not significant (P>0.05). There were no significant (P>0.05) differences found between the reproductive performance of sows in the two treatments; however this comparison was confounded by sows in the DM Area treatment having a lower number of matings per oestrus than those in the other treatment. Observations on the sexual behaviour of pigs at 145 matings indicated that the boars in the DM Area treatment displayed a higher (P<0.05) number of bouts of courting behaviour directed towards the female than boars in the conventional treatment (8.3 and 6.4, respectively). These very limited observations on sexual behaviour suggest that changes in the courting behaviour of bears may be associated with changes in litter size. While there was some indication from the results of progesterone analysis of blood samples taken from unmated gilts that there may have been some differences between treatments in the sexual age of gilts at matings, these differences are unlikely to explain the differences in litter size between treatments. Further research is required to identify the component (s) of the DM Area treatment that are responsible for the improvement in litter size in gilts and to further examine the effects of the two treatments on the reproductive performance of sows.     

Ammonia and other weak bases applied at any time of the hormone-dependent period inhibit 1-methyladenine-induced meiosis reinitiation of starfish oocytes

The effect of weak bases on starfish oocyte maturation induced by 1-methyladenine (1-MeAde) was investigated. Ammonium chloride, nicotine, and procaine inhibited meiosis reinitiation induced by either 1-MeAde or dithiothreitol. The inhibitory effect of the weak bases was highly dependent on the pH of sea water and increased with the increase in pH. These weak bases neither inhibited nor delayed meiosis reinitiation when given after the end of the hormone-dependent period, the period during which 1-MeAde is required for meiosis to occur, whereas they blocked hormone action when added just before the end of the hormone-dependent period.     

Genome Evolution of the Cyanobacterium Nostoc linckia under Sharp Microclimatic Divergence at "Evolution Canyon," Israel

We describe the genomic DNA diversity and divergence of the cyanobacterium Nostoc linckia from "Evolution Canyon," a microsite consisting of ecologically contrasting slopes, south-facing slope (SFS) and north-facing slope (NFS), at lower Nahal Oren, Mt. Carmel, Israel. The opposing slopes share their limestone lithology but vary greatly in their ecology, primarily because of different levels of solar radiation (which is six times higher on the SFS than on the NFS). The warm and xeric SFS displays a tropical African savanna, whereas the cool and mesic NFS displays a temperate South European Mediterranean live-oak maquis shrub forest. The cyanobacterium Nostoc linckia tested here is a sessile microorganism, growing as a carpet on rock surfaces and constantly exposed to environmental fluctuations of solar radiation, temperature, and desiccation. We demonstrate remarkable interslope and intraslope genetic divergence of the genome (including both coding and noncoding regions) of Nostoc linckia, by using 211 AFLP (amplified fragment length polymorphism) DNA molecular marker loci. Genetic polymorphism of N. linckia subpopulations on the ecologically harsher SFS was significantly (p <0.05) higher (p = 99.53%) than was that of the subpopulations on the climatically milder nfs (p = 85.78%). genetic polymorphism (p) and gene diversity (he) were significantly correlated with variables influencing aridity stress: solar radiation (sr) (rp = 0.956; p = 0.046), temperature (tm) (rp = 0.993; p = 0.0068), and day-night temperature difference (tdd) (rp = 0.975; p = 0.025). as in other tested organisms from "evolution canyon", but even more exceptionally because of its completely sedentary nature, we suggest that the climatically stressed sfs environment is responsible for this marked increase of genetic polymorphism, which is maintained by the combined evolutionary forces of diversifying and balancing selection. This could highlight the importance of ecological stress and selection in evolution and its remarkable effect on the genetic system across the prokaryotic genome.     

Mechanism of ferripyoverdine uptake by Pseudomonas aeruginosa outer membrane transporter FpvA: no diffusion channel formed at any time during ferrisiderophore uptake

To get access to iron, Pseudomonas aeruginosa produces the siderophore pyoverdine (PVD), composed of a fluorescent chromophore linked to an octapeptide, and its corresponding outer membrane transporter FpvA. This transporter is composed of three domains: a β-barrel inserted into the membrane, a plug that closes the channel formed by the barrel, and a signaling domain in the periplasm. The plug and the signaling domain are separated by a sequence of five residues called the TonB box, which is necessary for the interaction of FpvA with the inner membrane TonB protein. Genetic deletion of the plug domain resulted in the presence of a β-barrel in the outer membrane unable to bind and transport PVD-Fe. Expression of the soluble plug domain with the TonB box inhibited PVD-(55)Fe uptake most likely through interaction with TonB in the periplasm. A reconstituted FpvA in the bacterial outer membrane was obtained by the coexpression of separately encoded plug and β-barrel domains, each endowed with a signal sequence and a signaling domain. This resulted in polypeptide complementation after secretion across the cytoplasmic membrane. The reconstituted FpvA bound PVD-Fe with the same affinity as wild-type FpvA, indicating that the resulting transporter is correctly folded and reconstituted in the outer membrane. PVD-Fe uptake was TonB-dependent but 75% less efficient compared to wild-type FpvA. These data are consistent with a gated mechanism in which no open channel with a complete removal of the plug domain for PVD-Fe diffusion is formed in FpvA at any point during the uptake cycle.     

Microclimatic conditions, meltwater moistening, and the distributional pattern of Buellia frigida on rock in a southern continental Antarctic habitat

The importance of snowmelt as a source of moisture for the crustose lichen Buellia frigida in the early austral summer was investigated at Cape Geology, Granite Harbour, southern Victoria Land (77°01′S, 162°32′E). Surface and air temperatures and irradiance were recorded on the surface of a slightly inclined granite boulder for 5 weeks. Observations were made of lichen thallus hydration during a 5-day period. The results confirmed the strong warming effect of high irradiance; the rock surface and hydrated lichen were up to 19K above air temperature and, overall, the rock surface averaged 5.5K warmer. Therefore water condensation on the rock surface (dew or hoarfrost) was not possible during that period. Thalli were moistened by meltwater from both a small area of snow pack and from occasional snowfalls. The distribution of lichen thalli on the rock surface can be explained by the frequency and duration of meltwater moistening. Despite the very high irradiance whilst moist, the lichens seem well adapted to the combination of hydration, low temperatures and strong light. Received: 22 April 1997 / Accepted: 17 August 1997     

It's time to swim! Zebrafish and the circadian clock

The zebrafish represents a fascinating model for studying key aspects of the vertebrate circadian timing system. Easy access to early embryonic development has made this species ideal for investigating how the clock is first established during embryogenesis. In particular, the molecular basis for the functional development of the zebrafish pineal gland has received much attention. In addition to this dedicated clock and photoreceptor organ, and unlike the situation in mammals, the clocks in zebrafish peripheral tissues and even cell lines are entrainable by direct exposure to light thus providing unique insight into the function and evolution of the light input pathway. Finally, the small size, low maintenance costs and high fecundity of this fish together with the availability of genetic tools make this an attractive model for forward genetic analysis of the circadian clock. Here, we review the work that has established the zebrafish as a valuable clock model organism and highlight the key questions that will shape the future direction of research.