Dorsal and ventral stimuli in cell-material interactions: effect on cell morphology

Cells behave differently between bidimensional (2D) and tridimensional (3D) environments. While most of the in vitro cultures are 2D, most of the in vivo extracellular matrices are 3D, which encourages the development of more relevant culture conditions, seeking to provide more physiological models for biomedicine (e.g., cancer, drug discovery and tissue engineering) and further insights into any dimension-dependent biological mechanism. In this study, cells were cultured between two protein coated surfaces (sandwich-like culture). Cells used both dorsal and ventral receptors to adhere and spread, undergoing morphological changes with respect to the 2D control. Combinations of fibronectin and bovine serum albumin on the dorsal and ventral sides led to different cell morphologies, which were quantified from bright field images by calculating the spreading area and circularity. Although the mechanism underlying these differences remains to be clarified, excitation of dorsal receptors by anchorage to extracellular proteins plays a key role on cell behavior. This approach--sandwich-like culture--becomes therefore a versatile method to study cell adhesion in well-defined conditions in a quasi 3D environment.     

How successful are plant species reintroductions?

Reintroduction of native species has become increasingly important in conservation worldwide for recovery of rare species and restoration purposes. However, few studies have reported the outcome of reintroduction efforts in plant species. Using data from the literature combined with a questionnaire survey, this paper analyses 249 plant species reintroductions worldwide by assessing the methods used and the results obtained from these reintroduction experiments. The objectives were: (1) to examine how successful plant species reintroductions have been so far in establishing or significantly augmenting viable, self-sustaining populations in nature; (2) to determine the conditions under which we might expect plant species reintroductions to be most successful; (3) to make the results of this survey available for future plant reintroduction trials. Results indicate that survival, flowering and fruiting rates of reintroduced plants are generally quite low (on average 52%, 19% and 16%, respectively). Furthermore, our results show a success rate decline in individual experiments with time. Survival rates reported in the literature are also much higher (78% on average) than those mentioned by survey participants (33% on average). We identified various parameters that positively influence plant reintroduction outcomes, e.g., working in protected sites, using seedlings, increasing the number of reintroduced individuals, mixing material from diverse populations, using transplants from stable source populations, site preparation or management effort and knowledge of the genetic variation of the target species. This study also revealed shortcomings of common experimental designs that greatly limit the interpretation of plant reintroduction studies: (1) insufficient monitoring following reintroduction (usually ceasing after 4 years); (2) inadequate documentation, which is especially acute for reintroductions that are regarded as failures; (3) lack of understanding of the underlying reasons for decline in existing plant populations; (4) overly optimistic evaluation of success based on short-term results; and (5) poorly defined success criteria for reintroduction projects. We therefore conclude that the value of plant reintroductions as a conservation tool could be improved by: (1) an increased focus on species biology; (2) using a higher number of transplants (preferring seedlings rather than seeds); (3) taking better account of seed production and recruitment when assessing the success of reintroductions; (4) a consistent long-term monitoring after reintroduction.     

Universal criteria for species conservation priorities? Findings from a survey of public views across Europe

Decisions on biodiversity management and conservation are increasingly based on indicators. These imply, explicitly or implicitly, a number of criteria such as nativeness, rarity, endangeredness and (economic) value. We investigated to which degree such criteria matter to members of the general public and conducted a survey in eight sites across Europe (n = 2378). We explored the relationships between perceived desirability of a species’ population increase and six species-related attributes, including previous population change, rarity, vulnerability, harmfulness, value, attractiveness, and nativeness.For all three species types investigated, previous population change, followed by perceived harmfulness and value, had the strongest relationship with desirability of future increase. Perceived nativeness played only a minor role in informing a species’ desirability. A strong relationship between previous change and desirability of future increase could also be found in a number of additional species and six different habitat types, suggesting that previous change is a key criterion that the general public draws on to inform their attitudes towards biodiversity management.We compare the roles of such criteria for the general public to those used in the scientific and political discourse, and draw conclusions for the use of indicators in the conservation debate, arguing that biodiversity management that is strongly focused on nativeness might fall short of the interests of the citizenship, whereas other criteria, such as population trends, harmfulness and role and value of a species in the ecosystem strongly resonate with the views of the general public.     

Dynamic of nasal colonization by methicillin-resistant Staphylococcus aureus ST398 and ST1 after mupirocin treatment in a family in close contact with pigs

Nasal colonization by methicillin-resistant Staphylococcus aureus (MRSA) was evaluated after a mupirocin treatment in a family previously colonized by MRSA sequence type ST398 and ST1, who lived close to a pig farm. Eight nasal samples were swabbed from each of the four family members on different moments after mupirocin treatment. The efficacy of treatment was low in those family members who worked in the farm, and higher in the remaining two family members with sporadic contact with pigs. In addition, nasal and skin swabs from randomly selected pigs of the farm were taken. MRSA were detected in 33% of pigs tested. All MRSA isolates obtained were characterized by Staphylococcal-Cassette-Chromosome mec (SCCmec) determination, Multilocus-Sequence-Typing (MLST), spa- and agr-typing, Pulsed-field-gel-electrophoresis (PFGE), antimicrobial susceptibility, detection of antimicrobial resistance genes, and toxin gene profiling. Spa-types t011, t1255 and t1197 were detected in humans and animals, with indistinguishable PFGE patterns, suggesting animal to human MRSA transmission. Each spa-type was ascribed to a specific pulsotype. Spa-types t127 and t108 were only detected in MRSA isolates obtained from humans, and t012 only in those from animals. MRSA ST1-t127 isolates and some ST398-t011 and ST398-t1197 isolates presented a multiantimicrobial-resistance phenotype. None of them harbored lukF/lukS, tst, eta and etb virulence genes. This study showed that the efficacy of nasal MRSA decolonization in healthy people with very close contact with pigs is especially low.     

Molecular Characterization by AFLPs of Capsicum Germplasm from the Amazon Department in Colombia

Using AFLPs, 71 peppers (Capsicum) accessions from the species C. chinense Jacq., C. baccatum L., C. annuum L. and C. frutescens L. from indigenous communities of the Amazon Department (Colombia) were studied to assess the genetic diversity of the collections, and delineate species gene groups. Ten additional accessions were included as a reference species group. Three clusters were identified in the Amazonian accessions by Multiple Correspondence Analyses (MCA) and a dendrogram from the UPGMA analyses of Nei – Li genetic similarity. The clusters correspond to gene groups of the species Capsicum chinense (the majority of the accessions), C. baccatum and the complex annuum - frutescens. A fourth cluster corresponds to the reference accession C. pubescens. The MCA analyses accounted for 95% of the total variation. The total genetic variation was low (Ht 0.119) and a genetic diversity index (Gst) of 0.331 was obtained. This suggests a limited genetic diversity of the accessions and a close relatedness of the species. This study is the first molecular marker assessment of genetic diversity for peppers from the Colombian Amazon, and provides information of biodiversity that can be employed in the preservation and use of Capsicum germplasm.     

Progesterone and equine chorionic gonadotropin concentrations around the time of pregnancy loss in mares impregnated by donkeys or stallions

The objective of this study was to compare the incidence of pregnancy loss of mares carrying a mule embryo with that of mares carrying a horse embryo. The possible causes of such mortality were evaluated through serial ultrasonographic evaluations and hormonal monitoring, paying special attention to the role of premature regression of the endometrial cups and its relation to inadequate luteal function.

Twenty-eight mares impregnated by stallions and 19 mares impregnated with donkey semen were evaluated ultrasonographically every week from day 20 to day 150 of pregnancy. The viability of the product was assessed each time, and the diameter of the embryonic vesicle was measured from day 25 to day 60. Blood samples for progesterone and equine chorionic gonadotropin (eCG) determination were taken every week.

Both progesterone and eCG concentrations during normal pregnancies wegre significantly lower in the mares inseminated with donkey semen than in the mares impregnated by stallions (P<.05). In 7 of the mares carrying mule conceptuses to term, the concentrations of eCG remained basal throughout the study. In the other animals from this group, the levels of this hormone did increase but returned to baseline much earlier (on day 77 of pregnancy) than in the mares served by stallions (on day 126 day of pregnancy). There was no significant difference between the growth rate of embryonic vesicles of mares carrying mule embryos and that of mares carrying horse embryos (P>.05).

The incidence of pregnancy loss was significantly higher (P<.05) in mares carrying a mule embryo (36.8 %) than in mares carrying a horse embryo (21.4%); it occurred on average on day 93 of pregnancy in mares carrying mule embryos and on day 43 on mares carrying horse embryos. There was only 1 case in which pregnancy loss was associated with concentrations of both eCG and progesterone that were much lower than the average for the normal pregnancies of the same group, and this was in a mare carrying a horse embryo. The most frequent cause of pregnancy loss was premature luteal regression due to primary luteolysis, as evaluated via peripheral progesterone concentrations. This occurred in 2 mares carrying horse embryos and in 4 mares carrying mule embryos. Three mares carrying mule embryos and 1 carrying a horse embryo had abortions that were not preceded or accompanied by any alteration in progesterone or eCG levels and were thus classified as fetal deaths of non-endocrine origin.

It is concluded that the incidence of pregnancy loss is higher in mares carrying a mule embryo than in mares carrying a horse embryo. However, this is not due to the low progesterone concentrations associated with the premature regression of the endometrial cups that occurs in mares with interspecific pregnancy.     

Renal perfusion parameters measured by contrast‐enhanced ultrasound in healthy dogs demonstrate a wide range of variability in the long‐term

Contrast‐enhanced ultrasound may be helpful for detecting early renal microvascular damage and dysfunction in dogs. However, before this noninvasive imaging method can be tested as an early‐stage screening tool in clinical patients, an improved understanding of long‐term variation in healthy animals is needed. In this prospective, secondary, longitudinal, serial measurements study, variability of contrast‐enhanced ultrasound renal perfusion parameters was described for eight healthy dogs, using seven time points and a period of 83 weeks. Dogs were sedated with butorphanol (0.4 mg/kg), and contrast‐enhanced ultrasound of each kidney was performed after an intravenous bolus injection of a microbubble contrast agent (0.04 mL/kg). Time‐intensity curves were created from regions‐of‐interest drawn in the renal cortex and medulla. Intensity‐related parameters representing blood volume and time‐related parameters representing blood velocity were determined. A random‐effects model using restricted maximum likelihood was used to estimate variance components. Within‐dog coefficient of variation was defined as the ratio of the standard deviation over the mean. Time‐related parameters such as time‐to‐peak, rise and fall time had lowest within‐dog variability. Intensity‐related parameters such as peak enhancement, wash‐in and wash‐out area under the curve, total area under the curve, and wash‐in and washout rates had high within‐dog variability (coefficient of variation > 45%). Authors therefore recommend the use of time‐related parameters for future studies of renal perfusion. Within‐dog variability for bilateral kidney measurements was extremely low, therefore contrast‐enhanced ultrasound may be particularly useful for detecting unilateral changes in renal perfusion. Future studies are needed to compare contrast‐enhanced ultrasound findings in healthy dogs versus dogs with renal disease.     

Spatial and temporal variability of the two main caught species of an artisanal trap fishery in an oceanic island

1. This study analysed the catches of trap fishing in the Canary Archipelago, NE Atlantic Ocean. This study was conducted from October 2016 to September 2017, April to November 2018 and September 2020, in five ports (San Cristóbal, Taliarte, Castillo del Romeral, Arguineguín and Mogán).
2. The primary gear were traps with small mesh size (31.6 mm) and large mesh size (50.8 mm). A total of 2587 small mesh size traps and 141 large mesh size traps were analysed from 20- to 130-m depth.
3. The main target species were Dentex gibbosus and Stephanolepis hispidus.
4. Catches of traps with large mesh size showed D. gibbosus above first maturity and S. hispidus in small mesh size traps. Both species exhibit larger specimens during reproductive periods.
5. The most effective traps for D. gibbosus (large mesh) were located on the western coast (>30-m depth), with the highest catch per unit of effort (CPUE) in Mogán (857.7 g trap⁻¹ day⁻¹). The most effective traps for S. hispidus (small mesh) were located in southern areas, especially in sandy habitats (124.9 g trap⁻¹ day⁻¹ in Castillo del Romeral, 102.9 g trap⁻¹ day⁻¹ in Arguineguín).
6. The highest catchability of D. gibbosus was observed in April (CPUE of 962.8 g trap⁻¹ day⁻¹), and the highest catchability of S. hispidus was found in June (CPUE of 165.9 g trap⁻¹ day⁻¹).
7. The traps with small mesh size showed a great selectivity of sizes for S. hispidus; though high catch rates of immature specimens of D. gibbosus pose a threat to species conservation.
8. The management recommendations' proposed measures include revising minimum catch sizes for both species. Additionally, we suggested depth limitations for mesh sizes of traps.
9. The study highlights the need for new management strategies to ensure the conservation of targeted demersal species, with special emphasis on addressing the threat posed by small mesh sizes to immature D. gibbosus.
10. The findings provide valuable insights for adapting fishery management practices to sustainably conserve the targeted species in the Canary Archipelago.

     

Evaluation of dry ashing in conjunction with ion chromatographic determination of transition metal ions in pig feed samples

The contents of transition metal ions iron, copper, zinc, and manganese were simultaneously determined in pig feed using an ion chromatographic technique (IC) preceded by dry ashing. Employing ion exchange, the ions were separated on an IonPac CS5A column used in combination with a pyridine-2,6-dicarboxylic acid based eluent. The separation was followed by spectrophotometric detection after postcolumn reaction with 4-(2-pyridylazo)resorcinol. Dry ashing parameters were varied to assess their role in potential analyte loss. Quantitative recoveries (>95%) were obtained for all analytes with a dry ashing method that included a moderate temperature-time regime and ash leaching support in the form of sonication and heat treatment. The use of HCl as leaching acid and the presence of alkaline earths in the matrix solution did not interfere with the chromatographic separation.