Dispersive Raman spectroscopy allows the identification and quantification of melanin types

Melanins are the most prevalent pigments in animals and are involved in visual communication by producing colored traits that often evolve as intraspecific signals of quality. Identifying and quantifying melanins are therefore essential to understand the function and evolution of melanin‐based signals. However, the analysis of melanins is difficult due to their insolubility and the lack of simple methods that allow the identification of their chemical forms. We recently proposed the use of Raman spectroscopy as a simple, noninvasive technique that can be used to identify and quantify melanins in feathers and hairs. Contrarily, other authors later stated that melanins are characterized by a lack of defined Raman signals. Here, we use confocal Raman microscopy to confirm previous analyses showing that the two main chemical forms of melanins (eumelanin and pheomelanin) exhibit distinct Raman signal and compare different excitation wavelengths to analyze synthetic pheomelanin and natural melanins in feathers of different species of birds. Our analyses indicate that only laser excitation wavelengths below 1064 nm are useful for the analysis of melanins by Raman spectroscopy, and only 780‐nm laser in the case of melanins in feathers. These findings show that the capacity of Raman spectroscopy to distinguish different chemical forms of melanins depends on laser power and integration time. As a consequence, Raman spectroscopy should be applied after preliminar analyses using a range of these parameters, especially in fragile biological tissues such as feathers.     

Pathogenic bacteria and timing of laying

Pathogenic bacteria constitute a serious threat to viability of many organisms. Because growth of most bacteria is favored by humid and warm environmental conditions, earlier reproducers in seasonal environments should suffer less from the negative consequences of pathogenic bacteria. These relationships, and the effects on reproductive success, should be particularly prominent in predators because they are frequently exposed to pathogenic microorganisms from sick prey. Here, we presented and tested this hypothesis by sampling bacteria on adult and nestling goshawks Accipiter gentilis. We predicted that early breeders and their offspring should have fewer bacteria than those reproducing later during the breeding season. Adult goshawks with a high abundance of Staphylococcus on their beak and claws were easier to capture and their laying date was delayed. Moreover, goshawks that laid their eggs later had offspring with more Staphylococcus on their beaks and claws. The strength of the association between laying date and bacterial density of nestlings was stronger during the warm spring of 2013, when nestlings suffered from a higher abundance of pathogenic bacteria. Hatching failure and fledging failure were more common in nests with a higher abundance of Staphylococcus independently of the number of years occupied, laying date, and age of the female nest owner. These findings imply that timing of reproduction may be under the influence of pathogenic bacteria. Because early breeding goshawks produce more recruits than later breeders, our results suggest a role for pathogenic bacteria in the optimal timing of reproduction.     

A new Miocene local fauna from the Sierra Madre Oriental at San Luis Potosí, Central-East Mexico,and its paleontologic significance

Mexico's Late Neogene mammal faunas are largely known from localities in the Trans-Mexican Volcanic Belt; those from other morphotectonic provinces are few and far apart. Thus, the discovery of Late Miocene vertebrates in western Sierra Madre Oriental at San Luis Potosí, the Paso del Águila local fauna, significantly adds to this meager record. The assemblage was collected from the floodplain facies of the San Nicolás Formation, a ∼1100-m thick, dominantly fluviolacustrine and calcilithitic, 15°–20° NE dipping sequence preserved in the Peotillos-Tolentino Graben, between 22°11’–22°19’ N and 100°30’–100°39° W. It includes remains of cf. Trachemys, a small to medium-sized emydid chelonian, a large camelid, a small cervid and a new species of the equini Pliohippus s.s., comparable in size, cranial morphology and odontographic characters to the Clarendonian-Early Hemphillian horses of the Pliohippus clade. Ar-Ar dates from ash-fall tuffs seemingly above and below the fossiliferous strata, bracket the age between 12.33 and 7.41 Ma (i.e., late Middle to Late Miocene), that is, within the Late Clarendonian-Early Hemphillian NALMA interval, making this fauna the first in Mexico from this age. The Paso del Águila local fauna is at least partly correlative with the Hemphillian local faunas from the TMVB and adjacent areas (e.g., Rancho El Ocote, Guanajuato and Tecolotlán, Jalisco), the Central Plateau (e.g., Arroyo Los Fragmentos, Zacatecas), and the Sierra Madre Occidental (e.g., Yepómera). Elsewhere, it is broadly correlative with the Late Clarendonian-Early Hemphillian faunas from the California Coast Ranges (e.g., North Tejon Hills, Ricardo and Dove Springs in the Mohave Desert), and the Gulf Coast Plain, Florida (McGehee Farm and Mixon). The Paso del Águila local fauna was part of a subtropical savannah and pine-oak forest (with a well-developed understory) biome that thrived on a climate regime much more humid than today.     

Organogenesis and histological development of the wedge sole <Emphasis Type="Italic">Dicologoglossa cuneata</Emphasis> M. larva with special reference to the digestive system

In this work, several features during the wedge sole larval development have been described. The newly hatched larva presented an acidophilic yolk with some oil drops. The digestive tract began to differentiate at 1 DAH, with a loop being discernible. The pancreas and liver were completely formed at 2 DAH, the former showing its typical basophilic acinar structure and acidophilic zymogen granules. The first supranuclear vesicles in enterocytes were seen at 3 DAH. At 4 DAH, yolk reserves were completely exhausted, the number of oesophagus and intestine mucous cells increased, and the heart was differentiated into four chambers: the venous sinus, atrium, ventricle, and arterious bulb. The development was fast and almost all organs were differentiated at 2 DAH. It is important to emphasize that gastric glands were not detected, a factor that should be considered when deciding diet formulation and feeding strategies for the rearing of this species.     

Kinetic studies of Gly28:Ser mutant form of Bacillus pumilus lipase: Changes in kcat and thermal dependence

Lipases are useful catalysts for a wide variety of industrial purposes. Herein we report the stability and thermal dependence of the activity of wild-type Bacillus pumilus lipase (BplA) and four site-directed mutants designed to improve its thermal stability. The Gly28:Ser mutation produces a dramatic four-fold increase in its k_cat and a remarkable increase in its stability. While the increase in k_cat is temperature-independent, the increase in stability shows that the resultant interactions of this mutation have a strong enthalpic component. Thermal dependence of stability, k_cat, K_M and k_cat/K_M were analysed to gain insight on the structural effects of mutations on BplA. Our results are consistent with a gain in enzyme mobility for those mutants displaying enhanced catalytic properties; the analysis of thermal dependence of kinetic parameters indicates that the mutations did not change either the catalytic mechanism or the rate-limiting step of catalysis.     

Effects of relative humidity on development, fecundity and survival of three storage mites

The developmental rate of immature stages and the reproduction of adults of Tyrophagus putrescentiae (Schrank), T. neiswanderi Johnston and Bruce and Acarus farris (Oudemans) were examined at 70, 80 and 90% r.h. and a constant temperature of 25°C. At 70% r.h., T. putrescentiae and A. farris immature stages failed to reach the protonymph stage as 100% of the larvae died, whereas T. neiswanderi was able to complete development. The developmental time of all immature stages for the three species was significantly increased as relative humidity was reduced. The mobile stages were particularly susceptible, as the time needed to complete their development at lower relative humidities suffered greater increases than the egg stage. At 70% r.h., T. putrescentiae and A. farris were not able to lay eggs and only 24% of T. neiswanderi pairs were fertile. The reproductive parameters of the three species at the relative humidities at which they were able to lay eggs showed significant differences, except for the percentage of fertile mating at 80 and 90% r.h. As relative humidity increased, preoviposition period was reduced and fecundity and daily fecundity was increased, whereas the oviposition period showed different patterns for the three species. The intrinsic rate of increase (r _m ) of T. neiswanderi at 70% r.h. was negative indicating that, at these conditions, mite populations of this species will diminish until they disappear. As relative humidity increased from 80 to 90% r.h. this parameter was almost twofold for both Tyrophagus species. The r _m obtained for A. farris at 90% r.h. was similar to that of T. neiswanderi at the same humidity while at 80% r.h. it was very small so that the population doubling time was more than 84 days. The influence of relative humidity on biology of these mites and its practical application as control measure are discussed.     

Novel avenues for passion fruit in vitro regeneration from endosperm culture,and morpho-agronomic and physiological traits of triploid Passiflora cincinnata Mast. emblings

The present study describes a new regeneration system based on somatic embryogenesis from mature endosperm Passiflora cincinnata Mast. cultures. Moreover, the morpho-agronomic and phenological traits, as well as enzymatic activity of regenerated triploid emblings are compared to those of diploids. Mature endosperms were cultured on Murashige and Skoog medium supplemented with various concentrations (4.5–45.2 µM) of 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.5 μM 6-benzylaminopurine (BA). No plant growth regulators were included in the control group. Embryogenic calli were observed only in treatments supplemented with 13.6 and 18.1 µM 2,4-D?+?4.5 µM BA, with the highest number of somatic embryos per explant and regenerated plants (emblings) obtained with 18.1 µM 2,4-D. Most emblings (70%) were triploid (2n?=?3x?=?27), with a DNA amount (4.38 pg) similar to that of endosperm and 1.5 times greater than in diploid P. cincinnata seedlings (2n?=?2x?=?18), that contained 2.98 pg of DNA. While the number of organs and/or structures was akin to that in diploids, triploid emblings generally exhibited larger and longer vegetative and floral structures. The flower lifespan was also slightly altered by triploidy, nectar concentration was 27% higher, and the activity of plant defense enzymes β-1,3-glucanase and polyphenol oxidase was 29.8% and 22.1% higher. This study describes a new regeneration system for the production of phenotypic variants of this ornamental passion fruit species, opening new perspectives for future studies on genetic passion fruit breeding.

     

Gastrin-stimulated Gα13 Activation of Rgnef Protein (ArhGEF28) in DLD-1 Colon Carcinoma Cells

The guanine nucleotide exchange factor Rgnef (also known as ArhGEF28 or p190RhoGEF) promotes colon carcinoma cell motility and tumor progression via interaction with focal adhesion kinase (FAK). Mechanisms of Rgnef activation downstream of integrin or G protein-coupled receptors remain undefined. In the absence of a recognized G protein signaling homology domain in Rgnef, no proximal linkage to G proteins was known. Utilizing multiple methods, we have identified Rgnef as a new effector for Gα₁₃ downstream of gastrin and the type 2 cholecystokinin receptor. In DLD-1 colon carcinoma cells depleted of Gα₁₃, gastrin-induced FAK Tyr(P)-397 and paxillin Tyr(P)-31 phosphorylation were reduced. RhoA GTP binding and promoter activity were increased by Rgnef in combination with active Gα₁₃. Rgnef co-immunoprecipitated with activated Gα₁₃Q226L but not Gα₁₂Q229L. The Rgnef C-terminal (CT, 1279–1582) region was sufficient for co-immunoprecipitation, and Rgnef-CT exogenous expression prevented Gα₁₃-stimulated SRE activity. A domain at the C terminus of the protein close to the FAK binding domain is necessary to bind to Gα₁₃. Point mutations of Rgnef-CT residues disrupt association with active Gα₁₃ but not Gα_q. These results show that Rgnef functions as an effector of Gα₁₃ signaling and that this linkage may mediate FAK activation in DLD-1 colon carcinoma cells.