Regenerating vascular mural cells in zebrafish fin blood vessels are not derived from pre-existing mural cells and differentially require Pdgfrb signalling for their development.

Vascular networks comprise endothelial cells and mural cells, which include pericytes and smooth muscle cells. To elucidate the mechanisms controlling mural cell recruitment during development and tissue regeneration, we studied zebrafish caudal fin arteries. Mural cells colonizing arteries proximal to the body wrapped around them, whereas those in more distal regions extended protrusions along the proximo-distal vascular axis. Both cell populations expressed platelet-derived growth factor receptor ß (pdgfrb) and the smooth muscle cell marker myosin heavy chain 11a (myh11a). Most wrapping cells in proximal locations additionally expressed actin alpha2, smooth muscle (acta2). Loss of Pdgfrb signalling specifically decreased mural cell numbers at the vascular front. Using lineage tracing, we demonstrate that precursor cells located in periarterial regions and expressing Pgdfrb can give rise to mural cells. Studying tissue regeneration, we did not find evidence that newly formed mural cells were derived from pre-existing cells. Together, our findings reveal conserved roles for Pdgfrb signalling in development and regeneration, and suggest a limited capacity of mural cells to self-renew or contribute to other cell types during tissue regeneration.

Health Benefits of Consuming Foods with Bacterial Probiotics, Postbiotics, and Their Metabolites: A Review.

Over the years, probiotics have been extensively studied within the medical, pharmaceutical, and food fields, as it has been revealed that these microorganisms can provide health benefits from their consumption. Bacterial probiotics comprise species derived from lactic acid bacteria (LAB) (genus Lactobacillus, Leuconostoc, and Streptococcus), the genus Bifidobacterium, and strains of Bacillus and Escherichia coli, among others. The consumption of probiotic products is increasing due to the current situation derived from the pandemic caused by COVID-19. Foods with bacterial probiotics and postbiotics are premised on being healthier than those not incorporated with them. This review aims to present a bibliographic compilation related to the incorporation of bacterial probiotics in food and to demonstrate through in vitro and in vivo studies or clinical trials the health benefits obtained with their metabolites and the consumption of foods with bacterial probiotics/postbiotics. The health benefits that have been reported include effects on the digestive tract, metabolism, antioxidant, anti-inflammatory, anticancer, and psychobiotic properties, among others. Therefore, developing food products with bacterial probiotics and postbiotics is a great opportunity for research in food science, medicine, and nutrition, as well as in the food industry.

Microplastic concentration, distribution and dynamics along one of the largest Mediterranean-climate rivers: A whole watershed approach.

Microplastics (MPs) have been recognized as one of the most ubiquitous environmental pollutants globally. They have been found in all ecosystems studied to date, threatening biological diversity, ecosystem functioning and human health. The present study aimed to elucidate the environmental and anthropogenic drivers of MP dynamics in the whole catchment of the Biobío river, one of the largest rivers in South America. MP concentration and characteristics were analysed in 18 sites subjected to different sources of pollution and other human-related impacts. The sampling sites were classified in relation to altitudinal zones (highland, midland and lowland) and ecosystem types (fluvial and reservoir), and different water and territorial environmental variables were further collated and considered for analysis. Seven types of microplastic polymers were identified in the samples analysed, with a catchment mean (±SE) MP concentration of 22 ± 0.4 particles m-3, and MP presence being significantly higher in lowlands (26 ± 2 particle m-3) and in reservoirs (42 ± 14 particle m-3). The most abundant type of MP was fragments (84%), with a mean concentration of 37 ± 6 particles m-3. Overall, MP concentrations were low compared to those found in other studies, with a strong influence of human population size.

Monitoring of chromatin organization in live cells by FRIC. Effects of the inner nuclear membrane protein Samp1.

In most cells, transcriptionally inactive heterochromatin is preferentially localized in the nuclear periphery and transcriptionally active euchromatin is localized in the nuclear interior. Different cell types display characteristic chromatin distribution patterns, which change dramatically during cell differentiation, proliferation, senescence and different pathological conditions. Chromatin organization has been extensively studied on a cell population level, but there is a need to understand dynamic reorganization of chromatin at the single cell level, especially in live cells. We have developed a novel image analysis tool that we term Fluorescence Ratiometric Imaging of Chromatin (FRIC) to quantitatively monitor dynamic spatiotemporal distribution of euchromatin and total chromatin in live cells. A vector (pTandemH) assures stoichiometrically constant expression of the histone variants Histone 3.3 and Histone 2B, fused to EGFP and mCherry, respectively. Quantitative ratiometric (H3.3/H2B) imaging displayed a concentrated distribution of heterochromatin in the periphery of U2OS cell nuclei. As proof of concept, peripheral heterochromatin responded to experimental manipulation of histone acetylation. We also found that peripheral heterochromatin depended on the levels of the inner nuclear membrane protein Samp1, suggesting an important role in promoting peripheral heterochromatin. Taken together, FRIC is a powerful and robust new tool to study dynamic chromatin redistribution in live cells.

Mitotic spindle assembly and γ-tubulin localisation depend on the integral nuclear membrane protein Samp1.

We have investigated a possible role for the inner nuclear membrane protein Samp1 (also known as TMEM201) in the mitotic machinery. Live-cell imaging showed that Samp1a-YFP (Samp1a is the short isoform of Samp1) distributed as filamentous structures in the mitotic spindle, partially colocalising with ß-tubulin. Samp1 depletion resulted in an increased frequency of cells with signs of chromosomal mis-segregation and prolonged metaphase, indicating problems with spindle assembly and/or chromosomal alignment. Consistent with this, mitotic spindles in Samp1-depleted cells contained significantly lower levels of ß-tubulin and γ-tubulin, phenotypes that were rescued by overexpression of Samp1a-YFP. We found that Samp1 can bind directly to γ-tubulin and that Samp1 co-precipitated with γ-tubulin and the HAUS6 subunit of the Augmin complex in live cells. The levels of HAUS6, in the mitotic spindle also decreased after Samp1 depletion. We show that Samp1 is involved in the recruitment of HAUS6 and γ-tubulin to the mitotic spindle. Samp1 is the first inner nuclear membrane protein shown to have a function in mitotic spindle assembly.

Prevalence of inhibitors and clinical characteristics in patients with haemophilia in a middle-income Latin American country.

INTRODUCTION: Development of inhibitors is the most serious complication in patients with haemophilia (PWH). The prevalence of inhibitors in patients with severe haemophilia A (HA) is approximately 25%-30%. Inhibitor prevalence differs among populations. Some studies report a prevalence of almost twice in Hispanic as compared to Caucasian patients. Most data available, on the prevalence of inhibitors and their predisposing factors, originate from centres in developed countries. AIM: Establish the prevalence of inhibitors of FVIII and FIX in Mexico. METHODS: This was an observational, cross-sectional and descriptive study. The records of all patients diagnosed with haemophilia A (HA) or B (HB), with and without inhibitors, were included. Clinical and demographical characteristics of patients with inhibitors were assessed. Statistical analysis was performed using IBM SPSS version 22. The Ethics Committees of the various participating institutions approved this study. RESULTS: A total of 1455 patients from the 20 participating centres were recruited, from which 1208 (83.02%) had HA and 247 (16.97%) were diagnosed with HB. The presence of inhibitors in severe HA was reported in 93/777(11.96%), and 10/162 (6.17%) in severe HB. Of them, 91.7% exhibited high titres in HA and 100% in HB. CONCLUSION: In Mexico, the general prevalence of inhibitors varies considerably among centres. This study established a basis of comparison for future development and advances in the treatment and follow-up of patients. These findings also augment our understanding of risk factors related to inhibitor development.

Simulating rewetting events in intermittent rivers and ephemeral streams: A global analysis of leached nutrients and organic matter.

Climate change and human pressures are changing the global distribution and the extent of intermittent rivers and ephemeral streams (IRES), which comprise half of the global river network area. IRES are characterized by periods of flow cessation, during which channel substrates accumulate and undergo physico-chemical changes (preconditioning), and periods of flow resumption, when these substrates are rewetted and release pulses of dissolved nutrients and organic matter (OM). However, there are no estimates of the amounts and quality of leached substances, nor is there information on the underlying environmental constraints operating at the global scale. We experimentally simulated, under standard laboratory conditions, rewetting of leaves, riverbed sediments, and epilithic biofilms collected during the dry phase across 205 IRES from five major climate zones. We determined the amounts and qualitative characteristics of the leached nutrients and OM, and estimated their areal fluxes from riverbeds. In addition, we evaluated the variance in leachate characteristics in relation to selected environmental variables and substrate characteristics. We found that sediments, due to their large quantities within riverbeds, contribute most to the overall flux of dissolved substances during rewetting events (56%-98%), and that flux rates distinctly differ among climate zones. Dissolved organic carbon, phenolics, and nitrate contributed most to the areal fluxes. The largest amounts of leached substances were found in the continental climate zone, coinciding with the lowest potential bioavailability of the leached OM. The opposite pattern was found in the arid zone. Environmental variables expected to be modified under climate change (i.e. potential evapotranspiration, aridity, dry period duration, land use) were correlated with the amount of leached substances, with the strongest relationship found for sediments. These results show that the role of IRES should be accounted for in global biogeochemical cycles, especially because prevalence of IRES will increase due to increasing severity of drying events.

RanGTPase regulates the interaction between the inner nuclear membrane proteins, Samp1 and Emerin.

Samp1, spindle associated membrane protein 1, is a type II integral membrane protein localized in the inner nuclear membrane. Recent studies have shown that the inner nuclear membrane protein, Emerin and the small monomeric GTPase, Ran are direct binding partners of Samp1. Here we addressed the question whether Ran could regulate the interaction between Samp1 and Emerin in the inner nuclear membrane. To investigate the interaction between Samp1 and Emerin in live cells, we performed FRAP experiments in cells overexpressing YFP-Emerin. We compared the mobility of YFP-Emerin in Samp1 knock out cells and cells overexpressing Samp1. The results showed that the mobility of YFP-Emerin was higher in Samp1 knock out cells and lower in cells overexpressing Samp1, suggesting that Samp1 significantly attenuates the mobility of Emerin in the nuclear envelope. FRAP experiments using tsBN2 cells showed that the mobility of Emerin depends on RanGTP. Consistently, in vitro binding experiments showed that the affinity between Samp1 and Emerin is decreased in the presence of Ran, suggesting that Ran attenuates the interaction between Samp1 and Emerin. This is the first demonstration that Ran can regulate the interaction between two proteins in the nuclear envelope.

Molecular basis for the dual subcellular distribution of microsomal glutathione transferase 1.

Microsomal glutathione transferase 1 (MGST1) is a membrane bound enzyme involved in the detoxification of reactive electrophiles and protection of membranes from oxidative stress. The enzyme displays an unusual and broad subcellular distribution with especially high levels in the endoplasmic reticulum (ER) and outer mitochondrial membrane (OMM). Here we examined the molecular basis for this dual distribution. We hypothesized that the amphipathic properties of the first transmembrane segment (TMS), that contains a positively charged lysine (K25), is a central feature guiding dual targeting. The lysine-25 was substituted to alanine by site directed mutagenesis. We also increased the amphipathic character of the helix by inserting an additional lysine either one turn above or below K25. Expressing these constructs in simian COS cells, and analyzing subcellular distribution by immunocytochemistry, we observed an increased ER targeting of K25A-MGST1. In contrast I22K-MGST1 and F28K-MGST1 displayed pronounced mitochondrial targeting. By using in vitro transcription-translation we examined whether insertion of WT-MGST1 into ER is co- or post-translational and provide evidence for the former. In the same experimental set-up, mitochondrial insertion was shown to depend on the positive charge. Together these results show that removing the positive charge of lysine-25 promotes ER incorporation, but counteracts mitochondrial insertion. In contrast, introducing an extra lysine in the first TMS of MGST1 had opposite effects. The amphipathic character of the first TMS thus constitutes a molecular determinant for the dual targeting of MGST1. Broad subcellular distribution is consistent with a physiological role in protection from reactive intermediates and oxidative stress.

Saturated Fatty Acid Analogues of Cell-Penetrating Peptide PepFect14: Role of Fatty Acid Modification in Complexation and Delivery of Splice-Correcting Oligonucleotides.

Modifying cell-penetrating peptides (CPPs) with fatty acids has long been used to improve peptide-mediated nucleic acid delivery. In this study we have revisited this phenomenon with a systematic approach where we developed a structure-activity relationship to describe the role of the acyl chain length in the transfection process. For that we took a well-studied CPP, PepFect14, as the basis and varied its N-terminal acyl chain length from 2 to 22 carbons. To evaluate the delivery efficiency, the peptides were noncovalently complexed with a splice-correcting oligonucleotide (SCO) and tested in HeLa pLuc705 reporter cell line. Our results demonstrate that biological splice-correction activity emerges from acyl chain of 12 carbons and increases linearly with each additional carbon. To assess the underlying factors regarding how the transfection efficacy of these complexes is dependent on hydrophobicity, we used an array of different methods. For the functionally active peptides (C12-22) there was no apparent difference in their physicochemical properties, including complex formation efficiency, hydrodynamic size, and zeta potential. Moreover, membrane activity studies with peptides and their complexes with SCOs confirmed that the toxicity of the complexes at higher molar ratios is mainly caused by the free fraction of the peptide which is not incorporated into the peptide/oligonucleotide complexes. Finally, we show that the increase in splice-correcting activity correlates with the ability of the complexes to associate with the cells. Collectively these studies lay the ground work for how to design highly efficient CPPs and how to optimize their oligonucleotide complexes for lowest toxicity without losing efficiency.

Pollution Assessment of the Biobío River (Chile): Prioritization of Substances of Concern Under an Ecotoxicological Approach.

The water demand for human activities is rapidly increasing in developing countries. Under these circumstances, preserving aquatic ecosystems should be a priority which requires the development of quality criteria. In this study we perform a preliminary prioritization of the risky substances based on reported ecotoxicological studies and guidelines for the Biobío watershed (Central Chile). Our specific aims are (1) reviewing the scientific information on the aquatic pollution of this watershed, (2) determining the presence and concentration of potential toxic substances in water, sediment and effluents, (3) searching for quality criteria developed by other countries for the selected substances and (4) prioritizing the most risky substances by means of deterministic ecotoxicological risk assessment. We found that paper and mill industries were the main sources of point pollution, while forestry and agriculture were mostly responsible for non-point pollution. The most risky organic substances in the water column were pentachlorophenol and heptachlor, while the most relevant inorganic ones were aluminum, copper, unionized ammonia and mercury. The most risky organic and inorganic substances in the sediment were phenanthrene and mercury, respectively. Our review highlights that an important effort has been done to monitor pollution in the Biobío watershed. However there are emerging pollutants and banned compounds-especially in sediments-that require to be monitored. We suggest that site-specific water quality criteria and sediment quality criteria should be developed for the Biobío watershed, considering the toxicity of mixtures of chemicals to endemic species, and the high natural background level of aluminum in the Biobío.

Biotic and abiotic variables influencing plant litter breakdown in streams: a global study.

Plant litter breakdown is a key ecological process in terrestrial and freshwater ecosystems. Streams and rivers, in particular, contribute substantially to global carbon fluxes. However, there is little information available on the relative roles of different drivers of plant litter breakdown in fresh waters, particularly at large scales. We present a global-scale study of litter breakdown in streams to compare the roles of biotic, climatic and other environmental factors on breakdown rates. We conducted an experiment in 24 streams encompassing latitudes from 47.8° N to 42.8° S, using litter mixtures of local species differing in quality and phylogenetic diversity (PD), and alder (Alnus glutinosa) to control for variation in litter traits. Our models revealed that breakdown of alder was driven by climate, with some influence of pH, whereas variation in breakdown of litter mixtures was explained mainly by litter quality and PD. Effects of litter quality and PD and stream pH were more positive at higher temperatures, indicating that different mechanisms may operate at different latitudes. These results reflect global variability caused by multiple factors, but unexplained variance points to the need for expanded global-scale comparisons.

Anchoring of FRET Sensors-A Requirement for Spatiotemporal Resolution.

FRET biosensors have become a routine tool for investigating mechanisms and components of cell signaling. Strategies for improving them for particular applications are continuously sought. One important aspect to consider when designing FRET probes is the dynamic distribution and propagation of signals within living cells. We have addressed this issue by directly comparing an anchored (taFS) to a non-anchored (naFS) cleavable FRET sensor. We chose a microtubule-associated protein tau as an anchor, as microtubules are abundant throughout the cytosol of cells. We show that tau-anchored FRET sensors are concentrated at the cytoskeleton and enriched in the neurite-like processes of cells, providing high intensity of the total signal. In addition, anchoring limits the diffusion of the sensor, enabling spatiotemporally resolved monitoring of subcellular variations in enzyme activity. Thus, anchoring is an important aspect to consider when designing FRET sensors for deeper understanding of cell signaling.

MCLIP, an effective method to detect interactions of transmembrane proteins of the nuclear envelope in live cells.

Investigating interactions of proteins in the nuclear envelope (NE) using co-immunoprecipitation (Co-IP) has previously been difficult or even impossible due to their inherent resistance to extraction. We have developed a novel method, MCLIP (Membrane protein Cross-Link ImmunoPrecipitation), which takes advantage of a cell permeable crosslinker to enable effective detection and analysis of specific interactions of NE proteins in live cells using Western blot. Using MCLIP we show that, in U2OS cells, the integral inner nuclear membrane protein Samp1 interacts with Lamin B1, the LINC (Linker of nucleoskeleton and cytoskeleton) complex protein, Sun1 and the soluble small GTPase Ran. The results show that the previously detected in vitro interaction between Samp1 and Emerin also takes place in live cells. In vitro pull down experiments show, that the nucleoplasmic domains of Samp1 and Emerin can bind directly to each other. We also, show that MCLIP is suitable to coprecipitate protein interactions in different stages of the cell cycle.

Cost of Patients with Alzheimer's Disease in Spain According to Disease Severity.

BACKGROUND: Alzheimer's disease (AD) has been associated with great healthcare and non-healthcare resource consumption. The aim of this study was to estimate the burden of AD in Spain according to disease severity from a societal perspective. METHODS: A self-administered questionnaire was designed by the researchers and completed by the informal caregivers of patients with AD, reporting data on themselves as caregivers and on the AD patients for whom they care. The patients' sociodemographic and clinical data, their healthcare and non-healthcare resource consumption in the previous 12 months, and the impact of the disease on labor productivity were compiled. Data collected on informal caregivers included sociodemographic data and the impact of caring for a person with AD on their quality of life and labor productivity. Costs were estimated by multiplying the number of consumed resources by their unit prices. The cost of informal care was assessed using the proxy good method, and labor productivity losses were estimated using the human capital method. Costs were estimated by disease severity and are presented per patient per year in 2021 euros (€). RESULTS: The study sample comprised 171 patients with AD aged 79.1 ± 7.4 years; 68.8% were female, time from diagnosis was 5.8 ± 4.1 years, diagnosis delay was 1.8 ± 2.3 years, and the mean Cumulative Illness Rating Scale-Geriatric (CIRS-G) total was score 8.2 ± 6.0. According to disease severity, 14% had mild cognitive impairment or mild AD, 43.9% moderate AD, and 42.1% severe AD. The average annual cost per patient was €42,336.4 in the most conservative scenario. The greatest proportion of this cost was attributed to direct non-healthcare costs (86%, €36,364.8), followed by direct healthcare costs (8.6%, €3647.1), social care costs (4.6%, €1957.1), and labor productivity losses (less than 1%, €367.4). Informal care was the highest cost item, representing 80% of direct non-healthcare costs and 69% of the total cost. The total direct non-healthcare cost and total cost were significantly higher in moderate to severe disease severities, compared to milder disease severity. CONCLUSIONS: AD poses a substantial burden on informal caregivers, the national healthcare system, and society at large. Early diagnosis and treatment to prevent disease progression could reduce this economic impact.

Optimizing Lactic Acid Bacteria Proportions in Sourdough to Enhance Antifungal Activity and Quality of Partially and Fully Baked Bread.

The organic acids produced by lactic acid bacteria (LAB) during the fermentation of sourdoughs have the ability to reduce the growth of different molds. However, this ability depends on the LAB used. For this reason, in this study, the proportions of different LAB were optimized to obtain aqueous extracts (AEs) from sourdough to reduce fungal growth in vitro, control the acetic acid concentration, and obtain a specific lactic to acetic acid ratio. In addition, the optimized mixtures were used to formulate partially baked bread (PBB) and evaluate the mold growth and bread quality during refrigerated storage. Using a simplex-lattice mixture design, various combinations of Lactiplantibacillus plantarum, Lacticaseibacillus casei, and Lactobacillus acidophilus were evaluated for their ability to produce organic acids and inhibit mold growth. The mixture containing only Lpb. plantarum significantly reduced the growth rates and extended the lag time of Penicillium chrysogenum and P. corylophilum compared with the control. The AEs' pH values ranged from 3.50 to 3.04. Organic acid analysis revealed that using Lpb. plantarum yielded higher acetic acid concentrations than when using mixed LAB. This suggests that LAB-specific interactions significantly influence organic acid production during fermentation. The reduced radial growth rates and extended lag times for both molds compared to the control confirmed the antifungal properties of the AEs from the sourdoughs. Statistical analyses of the mixture design using polynomial models demonstrated a good fit for the analyzed responses. Two optimized LAB mixtures were identified that maximized mold lag time, targeted the desired acetic acid concentration, and balanced the lactic to acetic acid ratio. The addition of sourdough with optimized LAB mixtures to PBB resulted in a longer shelf life (21 days) and adequately maintained product quality characteristics during storage. PBB was subjected to complete baking and sensory evaluation. The overall acceptability was slightly higher in the control without sourdough (7.50), followed by bread formulated with the optimized sourdoughs (ranging from 6.78 to 7.10), but the difference was not statistically significant (p > 0.05). The sensory analysis results indicated that the optimization was used to successfully formulate a sourdough bread with a sensory profile closely resembling that of a nonsupplemented one. The designed LAB mixtures can effectively enhance sourdough bread's antifungal properties and quality, providing a promising approach for extending bread shelf life while maintaining desirable sensory attributes.

Utilization of Whey for Eco-Friendly Bio-Preservation of Mexican-Style Fresh Cheeses: Antimicrobial Activity of Lactobacillus casei 21/1 Cell-Free Supernatants (CFS).

Using whey, a by-product of the cheese-making process, is important for maximizing resource efficiency and promoting sustainable practices in the food industry. Reusing whey can help minimize environmental impact and produce bio-preservatives for foods with high bacterial loads, such as Mexican-style fresh cheeses. This research aims to evaluate the antimicrobial and physicochemical effect of CFS from Lactobacillus casei 21/1 produced in a conventional culture medium (MRS broth) and another medium using whey (WB medium) when applied in Mexican-style fresh cheese inoculated with several indicator bacteria (Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes). The CFSs (MRS or WB) were characterized for organic acids concentration, pH, and titratable acidity. By surface spreading, CFSs were tested on indicator bacteria inoculated in fresh cheese. Microbial counts were performed on inoculated cheeses during and after seven days of storage at 4 ± 1.0 °C. Moreover, pH and color were determined in cheeses with CFS treatment. Lactic and acetic acid were identified as the primary antimicrobial metabolites produced by the Lb. casei 21/1 fermentation in the food application. A longer storage time (7 days) led to significant reductions (p < 0.05) in the microbial population of the indicator bacteria inoculated in the cheese when it was treated with the CFSs (MRS or WB). S. enterica serovar Typhimurium was the most sensitive bacteria, decreasing 1.60 ± 0.04 log10 CFU/g with MRS-CFS, whereas WB-CFS reduced the microbial population of L. monocytogenes to 1.67 log10 CFU/g. E. coli and S. aureus were the most resistant at the end of storage. The cheese's pH with CFSs (MRS or WB) showed a significant reduction (p < 0.05) after CFS treatment, while the application of WB-CFS did not show greater differences in color (ΔE) compared with MRS-CFS. This study highlights the potential of CFS from Lb. casei 21/1 in the WB medium as an ecological bio-preservative for Mexican-style fresh cheese, aligning with the objectives of sustainable food production and guaranteeing food safety.

Samp1 is functionally associated with the LINC complex and A-type lamina networks.

The transmembrane inner nuclear membrane (INM) protein Samp1 is required for anchoring centrosomes near the nuclei. Using high-resolution fluorescence microscopy we show that Samp1 is distributed in a distinct and characteristic pattern in the nuclear envelope (NE), where it partially colocalizes with the LINC complex protein Sun1. By studying the localization of Samp1 deletion mutants and fusion proteins, we conclude that the cysteine-rich N-terminal half of Samp1 is nucleoplasmically exposed and is responsible for targeting to the INM. It contains four conserved CxxC motifs with the potential to form zinc fingers. The distribution of cysteine-to-alanine substitution mutants, designed to prevent zinc finger formation, showed that NE localization of Samp1 depends on intact CxxC motifs. Overexpression of Samp1 zinc finger mutants produced an abnormal dominant phenotype characterized by disrupted organization of a selective subset NE proteins, including emerin, Sun1, endogenous Samp1 and, in some cases, lamin A/C, but not lamin B, Sun2 or nucleoporins. Silencing of Samp1 expression showed that emerin depends on Samp1 for its correct localization in the NE. Our results demonstrate that Samp1 is functionally associated with the LINC complex protein Sun1 and proteins of the A-type lamina network.

The role of spatial processes and environmental determinants in microgeographic shell variation of the freshwater snail Chilina dombeyana (Bruguière, 1789).

Wildlife data often show spatial organization, demonstrating positive correlations either as a result of processes occurring over the landscape or due to the influence of spatially structured environmental variables. It is, thus, essential to consider non-random spatial structure when evaluating the underlying causes of biological variation. In this study, we analyzed the population structure of Chilina dombeyana shell morphology of 14 populations that are close geographically and belong to the same hydrographic basin. We utilized a variation partitioning approach to evaluate the importance of spatial processes, such as migration, acting over the landscape, and environmental characteristics, including habitat and hydrologic characteristics, and the occurrence of aquatic predators in promoting between population variation. Our results demonstrate spatially structured variation in C. dombeyana shell morphology, with populations living near each other having more similar shell sizes than populations living farther apart. The shell size variation partition indicated that both spatially structured environmental factors and genetic relationships resulting from migration or shared common ancestry may explain this pattern. Shell shape variation, in contrast, was found to be essentially under the influence of non-spatially structured environmental factors, with habitat and water characteristics accounting for about half of the total variation among populations. The large proportion of the variation in shell size that is spatially structured demonstrates that spatial structure on morphological traits might be strong and highlights the need to consider such phenomenon in intraspecific studies of phenotypic evolution.

Baseline element composition of foliose and fruticose lichens along the steep climatic gradient of SW Patagonia (Aisén Region, Chile).

Samples of foliose (Nephroma antarcticum) and fruticose (Usnea sp.) lichens were collected across a steep climatic and vegetation gradient in a remote, almost pristine region of SW Chilean Patagonia. Concentrations of major and trace elements in lichens from the rainforest were among the lowest ever reported worldwide for foliose and fruticose lichens and can be considered background levels for the region. The two lichen growth forms showed different elemental compositions mainly due to the greater capacity of foliose thalli to intercept elements from windborne and canopy-leached particles. The patterns of spatial variation in the chemical composition of lichens were effectively explained by statistical methods and reflected the different availability of wet and dry deposition along the steep climatic gradient. Baseline values established for N. antarcticum samples growing in temperate Nothofagus forests were therefore distinct from those of samples growing in more open, drier habitats. The fruticose Usnea sp. showed a higher affinity for atmophile Hg, low concentrations of lithophilic elements, and the same baseline composition whether from temperate forests or from dry, barren environments. The provided background and baseline values against which variations can be measured will be useful in the early detection of local or regional climatic and environmental change, especially in view of the planned construction of hydropower dams under the recently approved HidroAysén Project.