Functional Toll-like receptor 4 expressed in lactotrophs mediates LPS-induced proliferation in experimental pituitary hyperplasia

Toll like receptor 4 (TLR4) has been characterized for its ability to recognize bacterial endotoxin lipopolysaccharide (LPS). Considering that infections or inflammatory processes might contribute to the progression of pituitary tumors, we analyzed the TLR4 functional role by evaluating the LPS effect on lactotroph proliferation in primary cultures from experimental pituitary tumors, and examined the involvement of PI3K-Akt and NF-κB activation in this effect. In addition, the role of 17β-estradiol as a possible modulator of LPS-induced PRL cell proliferation was further investigated. In estrogen-induced hyperplasic pituitaries, LPS triggered lactotroph cell proliferation. However, endotoxin failed to increase the number of lactotrophs taking up BrdU in normal pituitaries. Moreover, incubation with anti-TLR4 antibody significantly reduced LPS-induced lactotroph proliferation, suggesting a functional role of this receptor. As a sign of TLR4 activation, an LPS challenge increased IL-6 release in normal and tumoral cells. By flow cytometry, TLR4 baseline expression was revealed at the plasma membrane of tumoral lactotrophs, without changes noted in the percentage of double PRL/TLR4 positive cells after LPS stimulus. Increases in TLR4 intracellular expression were detected as well as rises in CD14, p-Akt and NF-κB after an LPS challenge, as assessed by western blotting. The TLR4/PRL and PRL/NF-κB co-localization was also corroborated by immunofluorescence and the involvement of PI3K/Akt signaling in lactotroph proliferation and IL-6 release was revealed through the PI3K inhibitor Ly-294002. In addition, 17β-estradiol attenuated the LPS-evoked increase in tumoral lactotroph proliferation and IL-6 release. Collectively these results demonstrate the presence of functional TLR4 in lactotrophs from estrogen-induced hyperplasic pituitaries, which responded to the proliferative stimulation and IL-6 release induced by LPS through TLR4/CD14, with a contribution of the PI3K-Akt and NF-κB signaling pathways.     

Effect of Proline Mutations on the Monomer Conformations of Amylin

The formation of human islet amyloid polypeptide (hIAPP) is implicated in the loss of pancreatic β-cells in type II diabetes. Rat amylin, which differs from human amylin at six residues, does not lead to formation of amyloid fibrils. Pramlintide is a synthetic analog of human amylin that shares three proline substitutions with rat amylin. Pramlintide has a much smaller propensity to form amyloid aggregates and has been widely prescribed in amylin replacement treatment. It is known that the three prolines attenuate β-sheet formation. However, the detailed effects of these proline substitutions on full-length hIAPP remain poorly understood. In this work, we use molecular simulations and bias-exchange metadynamics to investigate the effect of proline substitutions on the conformation of the hIAPP monomer. Our results demonstrate that hIAPP can adopt various β-sheet conformations, some of which have been reported in experiments. The proline substitutions perturb the formation of long β-sheets and reduce their stability. More importantly, we find that all three proline substitutions of pramlintide are required to inhibit β conformations and stabilize the α-helical conformation. Fewer substitutions do not have a significant inhibiting effect.     

Pollen production in the Poaceae family

Total pollen production per inflorescence was studied in the most important species of the Poaceae family in the city of Córdoba in order to further our knowledge of the partial contribution of each species of this family to the total amount of pollen released into the atmosphere.

The contribution of grasses in a given area was estimated by counting the number of inflorescences in an area of one square meter. Four different representative areas of the city were selected. The number of pollen grains per anther and flowers per inflorescence was also estimated in order to obtain total pollen production per inflorescence.

Pollen production per inflorescence ranged from 14,500 to more than 22,000,000 pollen grains, the amount being clearly higher in the perennial species. Pollen production per square meter was higher in the mountains near the city and lower in areas of abandoned crops.

Only a few species are responsible for the majority of pollen produced. A phenological study is necessary in order to determine the temporal distribution of this pollen production and subsequent shedding.     

Pollen biology in four Mediterranean Quercus species

Potential pollen production, viability and germination were studied in the most important species of Quercus in the mountains of Córdoba to determine the contribution of each species to the total amount of airborne pollen. The results were compared over two consecutive years with different rainfall patterns. The viability of pollen grains was determined at anther opening, and during the pollination period, in order to determine potential pollination capacity. Results indicated that there were differences in the number of pollen grains produced by stamen in the four species. Equally, there were differences in the number of flowers among the species, being Q. suber the species with higher number of catkins groups and flowers per individual tree. Total pollen production per tree can be summarized in the following proportion: 1: 3: 3: 6 (Q. coccifera, Q. ilex ssp. ballota, Q. faginea and Q. suber). Potential pollen viability was estimated using the Fluorochromatic Reaction (FCR) and a germination assay. The results have shown that Quercus potential pollen viability is high and declines slowly with time. Q. coccifera was the species with the highest percentage of germination, with Q. suber being the lowest.     

Agroecosystems and conservation of migratory waterbirds: importance of coastal pastures and factors influencing their use by wintering shorebirds

Coastal pastures are common agroecosystems adjacent to estuarine areas that can provide valuable habitat for wildlife, particularly for migratory shorebirds. Disentangling the factors that influence coastal pasture use by wintering shorebirds will provide new insights into its role for buffering human disturbances and habitat loss in intertidal areas. We examined whether numbers of two shorebirds (Eurasian curlew and Black-tailed godwit) foraging actively on coastal pastures was affected by weather conditions, tidal stage (low/high tide) and number of harvesters at intertidal areas throughout winter. Both species frequently used coastal pastures and most individuals foraged actively there. The average percentage of the total wintering population of curlews and godwits foraging on coastal pastures was 27.4 and 7.8 %, respectively, and was significantly higher during high tide compared to low tide. The number of harvesters on mudflats also had a positive significant effect in explaining the presence of curlews, and to a lesser extent for godwits, on coastal pastures, and accumulated rainfall had a positive effect for both species too. These supratidal areas were consistently used as alternative foraging grounds during low tide by curlews, as well as supplementary foraging areas during high tide by wintering populations of both large shorebirds. By supplementary foraging, wintering curlews, and probably godwits, seemed to compensate for a negative effect of the presence of harvesters on their foraging activity. We recommend managing of those coastal agricultural fields adjacent to intertidal foraging grounds in order to increase the availability of supratidal foraging habitats for declining shorebird populations. These habitats may thus have a beneficial role in sustaining populations of wintering shorebirds, but further studies are needed to estimate if birds can compensate for any shortfall in daily energy budget by supplementary foraging on coastal pastures, thus providing insights into whether they are involved in large-scale population regulation of migratory birds.     

PEG conjugates of potent α4 integrin inhibitors,maintaining sustained levels and bioactivity in vivo,following subcutaneous administration

Mitsunobu reactions were employed to link t-butyl esters of α4 integrin inhibitors at each of the termini of a three-arm, 40 kDa, branched PEG. Cleavage of the t-butyl esters using HCO₂H provided easily isolated PEG derivatives, which are potent α4 integrin inhibitors, and which achieve sustained levels and bioactivity in vivo, following subcutaneous administration to rats.     

Effects of cytoprotective antioxidants on lymphocytes from representative mitochondrial neurodegenerative diseases

Two new aza analogues of the neuroprotective agent idebenone have been synthesized and characterized. Their antioxidant activity, and ability to augment ATP levels have been evaluated in several different cell lines having suboptimal mitochondrial function. Both compounds were found to be good ROS scavengers, and to protect the cells from oxidative stress induced by glutathione depletion. The compounds were more effective than idebenone in neurodegenerative disease cells. These novel pyrimidinol derivatives were also shown to augment ATP levels in coenzyme Q₁₀-deficient human lymphocytes. The more lipophilic side chains attached to the pyrimidinol redox core in these compounds resulted in less inhibition of the electron transport chain and improved antioxidant activity.     

Crops and fodder: evidence for storage and processing activities in a functional area at the Roman settlement of Monte Mozinho (northern Portugal)

Archaeobotanical material was recovered in two contiguous compartments of a compound in Monte Mozinho (Penafiel, northern Portugal). These areas comprise storage facilities from three different typologies—dolium, pits and above-ground quadrangular structures in the corner of the compartments. Few carpological materials were found inside the storage structures and all these are cereals which were most likely stored as clean grain. Recovered grains are probably remnants of the last stored crops prior to abandonment of the area. A radiocarbon date obtained on grains of rye positions this event at a time within the 3rd century and the beginning of the 4th century a.d. Outside the structures from compartment 2, abundant crop processing by-products, mostly weeds and chaff, were recovered. Since traditional agricultural communities tend not to waste such organic material, it is likely that it was kept in the area to be used as fodder. Data is ambiguous regarding the possible existence of fodder crops mixed with these by-products. Overall, there is a predominance of millets (Panicum miliaceum and Setaria italica) and rye (Secale cereale), but naked wheat (Triticum aestivum) is also a relevant crop. The presence of oat (Avena) is significant but it is difficult to interpret due to its exclusive presence alongside weeds and chaff. Grape pips (Vitis vinifera) were also found, but these are rare. The Late Roman carpological material from Monte Mozinho is of great relevance on a regional level. It documents and dates the presence of rye and testifies to a multifunctional system in which plant husbandry and animal breeding were intertwined.     

Comparing galling insect richness among Neotropical savannas: effects of plant richness, vegetation structure and super-host presence

Plant species diversity maintains the stability of ecosystems and the diversity of consumer species such as insect herbivores. Considering that gall-inducing insects are highly specialized on their host plants and dependent on the occurrence, abundance and distribution of plants, we evaluated the diversity patterns of gall-inducing insect along Brazilian Neotropical savannas and the potential role of plant species richness, vegetation structure and super-host presence on determining these patterns. We found 1,882 individual plants that belonged to 64 different host plant species grouped in 31 families, associated to 112 galling insect species. The galling richness was positively influenced by plant species richness and the presence of the super-host genus Qualea (Vochysiaceae). Plant species richness explained 48 % of the galling richness and areas with presence of super-hosts had more than twice of galling species than areas where they were absent. On the other hand, we found no evidence that larger plants hosted more species of galling insects. We observed that for the diversity of galling insects in the Brazilian Cerrado, vegetation structure explained almost the same portion as plant richness, because structural variables did not have an effect on residuals of galling richness and plant richness regression. Our findings suggests that plant richness has a more important role on the mitigation of natural enemies and adaptive radiation of galling species, while structural aspects of the vegetation does not seem to have that effect. Furthermore, we show that the super-host taxa provide an increment in local galling richness because they present a great diversity of local number of gall morphospecies (i.e. alpha diversity) and the high turnover of morphospecies among different localities (i.e. beta diversity). Therefore we argue that the quality of resources (richness and super host presence) appears to be a most important factor for the diversity of galling insects in Neotropical systems, than the amount of resources.