Prostaglandins involvement in the formation of luteinized unrupted follicles in the cyclic female rat

The purpose of this investigation was to study the role played by prostaglandins in advanced ovulation and in the formation of luteinized unrupted follicles (LUF) in cyclic female rats. Dose related effects on ovulation were observed in rats given LH on diestrus 2 at 16.30. A significant positive correlation was observed between the number of postovulatory corpura lutea (POCL) and the increasing doses of LH. By contrast the number of LUF was negatively correlated with LH. Indomethacin treatment by 6h30 after administration of an ovulatory LH dose significantly increased the occurence of LUF at the expense of POCL. Conversely PGF_2α when admiststered by 6h30 after a subovulatory LH stimulation enhanced in a dose dependent manner the number of POCL with respect to the LH treated controls. Under a similar treatment with a subovulatory dose of LH, PGE₂ remained without ovulatory effects. The mechanisms of the formation of LUF are discussed on the basis of these results.     

Hydraulic acclimation in a Mediterranean oak subjected to permanent throughfall exclusion results in increased stem hydraulic capacitance

Stem water storage capacity and hydraulic capacitance (C_S) play a crucial role in tree survival under drought-stress. To investigate whether C_S adjusts to increasing water deficit, variation in stem water content (StWC) was monitored in vivo for 2 years and related to periodical measurements of tree water potential in Mediterranean Quercus ilex trees subjected either to permanent throughfall exclusion (TE) or to control conditions. Seasonal reductions in StWC were larger in TE trees relative to control ones, resulting in greater seasonal C_S (154 and 80 kg m⁻³ MPa⁻¹, respectively), but only during the first phase of the desorption curve, when predawn water potential was above −1.1 MPa. Below this point, C_S decreased substantially and did not differ between treatments (<20 kg m⁻³ MPa⁻¹). The allometric relationship between tree diameter and sapwood area, measured via electrical resistivity tomography, was not affected by TE. Our results suggest that (a) C_S response to water deficit in the drought-tolerant Q. ilex might be more important to optimize carbon gain during well-hydrated periods than to prevent drought-induced embolism formation during severe drought stress, and (b) enhanced C_S during early summer does not result from proportional increases in sapwood volume, but mostly from increased elastic water.     

Autosomal dominant polycystic kidney disease: evidence for the existence of a third locus in a Portuguese family

Autosomal dominant polycystic kidney disease is characterized by clinical and genetic heterogeneity. Two loci implicated in the disease have previously been mapped (PKD1 on chromosome 16 and PKD2 on chromosome 4). By two point and multipoint linkage analysis, negative lod scores have been found for both chromosome 16 and chromosome 4 markers in a large Portuguese family, indicating that a third PKD locus is involved in the development of the disease.     

Circadian clock adjustment to plant iron status depends on chloroplast and phytochrome function

Plant chloroplasts are not only the main cellular location for storage of elemental iron (Fe), but also the main site for Fe, which is incorporated into chlorophyll, haem and the photosynthetic machinery. How plants measure internal Fe levels is unknown. We describe here a new Fe‐dependent response, a change in the period of the circadian clock. In Arabidopsis, the period lengthens when Fe becomes limiting, and gradually shortens as external Fe levels increase. Etiolated seedlings or light‐grown plants treated with plastid translation inhibitors do not respond to changes in Fe supply, pointing to developed chloroplasts as central hubs for circadian Fe sensing. Phytochrome‐deficient mutants maintain a short period even under Fe deficiency, stressing the role of early light signalling in coupling the clock to Fe responses. Further mutant and pharmacological analyses suggest that known players in plastid‐to‐nucleus signalling do not directly participate in Fe sensing. We propose that the sensor governing circadian Fe responses defines a new retrograde pathway that involves a plastid‐encoded protein that depends on phytochromes and the functional state of chloroplasts.     

Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle

Trypanosoma cruzi, etiological agent of Chagas’ disease, has a complex life cycle which involves the invasion of mammalian host cells, differentiation and intracellular replication. Here we report the first insights into the biological role of a poly(ADP-ribose) glycohydrolase in a trypanosomatid (TcPARG). In silico analysis of the TcPARG gene pointed out the conservation of key residues involved in the catalytic process and, by Western blot, we demonstrated that it is expressed in a life stage-dependant manner. Indirect immunofluorescence assays and electron microscopy using an anti-TcPARG antibody showed that this enzyme is localized in the nucleus independently of the presence of DNA damage or cell cycle stage. The addition of poly(ADP-ribose) glycohydrolase inhibitors ADP-HPD (adenosine diphosphate (hydroxymethyl) pyrrolidinediol) or DEA (6,9-diamino-2-ethoxyacridine lactate monohydrate) to the culture media, both at a 1 µM concentration, reduced in vitro epimastigote growth by 35% and 37% respectively, when compared to control cultures. We also showed that ADP-HPD 1 µM can lead to an alteration in the progression of the cell cycle in hydroxyurea synchronized cultures of T. cruzi epimastigotes. Outstandingly, here we demonstrate that the lack of poly(ADP-ribose) glycohydrolase activity in Vero and A549 host cells, achieved by chemical inhibition or iRNA, produces the reduction of the percentage of infected cells as well as the number of amastigotes per cell and trypomastigotes released, leading to a nearly complete abrogation of the infection process. We conclude that both, T. cruzi and the host, poly(ADP-ribose) glycohydrolase activities are important players in the life cycle of Trypanosoma cruzi, emerging as a promising therapeutic target for the treatment of Chagas’ disease.     

DIATMOD: diatom predictive model for quality assessment of Portuguese running waters

A predictive model for diatoms based on an adaptation of the River Invertebrate Prediction and Classification System/Australian River Assessment System approaches was evaluated as an effective tool for measuring stream ecological quality. This type of model was originally developed in UK and later in Australia and is extensively used to obtain ecological quality assessments with macroinvertebrates. The first step for the model construction was the definition of six consistent reference biological groups (ANOSIM: Global R = 0.77; P < 0.001) after classification (UPGMA) and ordination (nMDS) of 120 reference sites containing 254 different diatom taxa (species and infra-specific rank). A set of five environmental variables (slope, hydrological regime, mean annual temperature, mean annual precipitation and alkalinity) correctly discriminated 67% of reference sites (stepwise forward discriminant analysis, Jackknifed classification). The model was statistically accurate (slope = 1.07, intercept = −0.68, R ² = 0.65) and was validated by an independent set of reference data (13 reference sites; 70% correct answers). In addition, the model was tested by running data from 113 potentially disturbed sites. The model (DIATMOD) was well correlated with a general abiotic degradation gradient (Spearman correlations, R ² = 0.53, P < 0.001; and PCA analysis) and also with several specific pressure variables such as nitrates, phosphates, urban area, connectivity and land use (P < 0.001). Most diatom indices assess chemical contamination and we showed here that through predictive modelling the potential of diatoms as bioindicators increases as they also responded to hydromorphological changes. Further investigation on model potential consists in: testing different probability levels for taxa inclusion (here it was >0.5 as the most common models); comparing with alternative classification systems; assessing the influence of substrate type and seasonal variation in assessments.     

The first record of Lutzomyia longipalpis in the Argentine northwest

In 2004, the urban presence of Lutzomyia longipalpis was recorded for the first time in Formosa province. In 2006, the first autochthonous case of human urban visceral leishmaniasis (VL) was recorded in Misiones in the presence of the vector, along with some canine VL cases. After this first case, the vector began to spread primarily in northeast Argentina. Between 2008-2011, three human VL cases were reported in Salta province, but the presence of Lu. longipalpis was not recorded. Captures of Phlebotominae were made in Tartagal, Salta, in 2013, and the presence of Lu. longipalpis was first recorded in northwest Argentina at that time. Systematic sampling is recommended to observe the distribution and dispersion patterns of Lu. longipalpis and consider the risk of VL transmission in the region.     

Intracellular Nucleic Acid Delivery by the Supercharged Dengue Virus Capsid Protein

Supercharged proteins are a recently identified class of proteins that have the ability to efficiently deliver functional macromolecules into mammalian cells. They were first developed as bioengineering products, but were later found in the human proteome. In this work, we show that this class of proteins with unusually high net positive charge is frequently found among viral structural proteins, more specifically among capsid proteins. In particular, the capsid proteins of viruses from the Flaviviridae family have all a very high net charge to molecular weight ratio (> +1.07/kDa), thus qualifying as supercharged proteins. This ubiquity raises the hypothesis that supercharged viral capsid proteins may have biological roles that arise from an intrinsic ability to penetrate cells. Dengue virus capsid protein was selected for a detailed experimental analysis. We showed that this protein is able to deliver functional nucleic acids into mammalian cells. The same result was obtained with two isolated domains of this protein, one of them being able to translocate lipid bilayers independently of endocytic routes. Nucleic acids such as siRNA and plasmids were delivered fully functional into cells. The results raise the possibility that the ability to penetrate cells is part of the native biological functions of some viral capsid proteins.