Characterization of tumor-associated lymphocytes in a series of mouse mammary tumor lines with differing biological properties

Tumor-associated lymphocytes were isolated by isokinetic gradient separation from five related mouse mammary tumor lines with different immunological and growth characteristics. Although considerable variation in recovery rates was seen from experiment to experiment, the five tumor types were found to have reproducible and characteristic patterns of T lymphocyte subpopulations, as detected by cytotoxicity assay using monoclonal antisera to Thy-1, Lyt-1, and Lyt-2 antigens. Tumors of line 168, which are weakly immunogenic at best, had the lowest numbers of recovered ALS+, Thy1+ lymphocytes (12% and 9%, respectively), in contrast to immunogenic lines (mean 38% and 26%, respectively). Line 68H tumors, which grow after prolonged latency periods and also produce tumor cell variants in vivo, were unique in that the numbers of recovered Lyt 1+ lymphocytes exceeded the number of Lyt 2+ lymphocytes, whereas these two T cell subpopulations were either equal or Lyt 2+ cells predominated in the other faster growing, non-variant-producing tumors. No differences in T lymphocyte distribution were associated with the presence or absence of metastatic behavior. These results indicate that distinctive lymphocyte infiltrates may be characteristic of tumors with distinct biological differences.     

Effect of vertebrate feces on the attraction of Culicoides (Diptera: Ceratopogonidae) in an Amazonian mangrove

Biting midges are of interest to public health because they play an important role as vectors of disease‐causing pathogens, as well as being a biting nuisance to humans and domestic animals. Although these insects are common in mangrove areas, they have not yet been studied in this ecosystem in the state of Maranhão (MA), Brazil. The objective of this study was to characterize the Culicoides community structure found in a mangrove swamp and verify if the use of vertebrate feces as bait interferes with their composition, richness, abundance, or seasonality. CDC light traps with vertebrate (chicken, ox, control, donkey, capuchin monkey and pig) feces baits were used to capture biting midges in a mangrove area of the Island of São Luís, MA. A total of 4,087 individuals representing 22 species of Culicoides were captured, the most abundant being C. (Oecacta) furens Poey (23.46%), C. (gr. Fluviatilis) leopoldoi Ortiz (21.58%), C. (Hoffmania) ignacioi Forattini (16.98%), C. (Hoffmania) maruim Lutz (13.85%), C. (Diphaomyia) iriartei Fox (10.57%), C. (Hoffmania) insignis Lutz (7.07%), and C. (gr. Limai) limai Barreto (3.03%). Species richness and abundance were higher when baits of capuchin monkey (15 species; 26.84% of the individuals) and pig (15; 25.3%) feces were used. The least attractive baits were donkey (12 species; 9.3%) and ox (nine species; 11.52%) feces. Biting midges were more abundant in the rainy season (67%), but richness was higher in the dry season (19 species). These results show that vertebrate feces may serve as olfactory cues and increase the attraction of biting midges to traps.     

Pelage color variation in pocket gophers (Rodentia: Geomyidae) in relation to sex,age and differences in habitat

Pelage coloration is a phenotypic characteristic in mammals that could be associated with an individual's survival and fitness; thus coloration gains adaptive importance. In geomyids, pelage coloration shows a relationship with the color of soils on which they live, mainly freshly dug soil of their burrows. This characteristic could be due to a camouflage adaptation to avoid predators. Pocket gophers disperse aboveground to establish a territory before they reach reproductive condition, and males disperse longer distances than females. The aim of this study was to evaluate pelage color variation in pocket gophers (Thomomys anitae) in relation to habitat differences, sex, and age, and determine its association to the color of the soil on which they live. Brightness of T. anitae's dorsal pelage coloration and that of soil samples from five different habitats in the Baja California peninsula, Mexico were measured to test four hypotheses: (1) Subadults show a wide coloration range, but extreme colors are lost in adulthood due to natural selection. (2) Males are more vulnerable to depredation than females; therefore, males’ coloration is more homogenous as a protective camouflage. (3) In open habitats pocket gophers are more exposed to being detected by predators, therefore their pelage coloration pattern is less variable than that of individuals from habitats with more vegetation cover. (4) Pelage coloration better matches soil coloration in moist conditions similar to that of freshly dug soil of their burrows. The results confirmed our predictions; however, selection does not impose an equal pressure on pelage coloration on the five habitats evaluated; other factors such as population density and predator presence need to be assessed. The strongest effects are found in the most open habitat, and there is less strong support for predictions in habitats where predator assemblage is diminished.     

Golgi localisation of GMAP210 requires two distinct cis-membrane binding mechanisms

Background  

The Golgi apparatus in mammals appears as a ribbon made up of interconnected stacks of flattened cisternae that is positioned close to the centrosome in a microtubule-dependent manner. How this organisation is achieved and retained is not well understood. GMAP210 is a long coiled-coil cis-Golgi associated protein that plays a role in maintaining Golgi ribbon integrity and position and contributes to the formation of the primary cilium. An amphipathic alpha-helix able to bind liposomes in vitro has been recently identified at the first 38 amino acids of the protein (amphipathic lipid-packing sensor motif), and an ARF1-binding domain (Grip-related Arf-binding domain) was found at the C-terminus. To which type of membranes these two GMAP210 regions bind in vivo and how this contributes to GMAP210 localisation and function remains to be investigated.     

Placental morphology in two sympatric Andean lizards of the genus Liolaemus (Reptilia: Liolaemidae)

Viviparity is a remarkable feature in squamate sauropsids and it has evolved multiple times in parallel with the formation of a placenta. One example of this repeated evolution of viviparity and placentation occurs in the species‐rich South American genus Liolaemus with at least six independent origins of viviparity. However, evolutionary studies of placentation in this genus are limited by a lack of data on placental morphology. The aim of this study is to describe and compare the microanatomy and vessel diameter (Dv, a function of blood flow) of the placenta using scanning electron microscopy (SEM) and confocal laser scanning microscopy (cLSM) in two sympatric Andean viviparous but highly divergent species, Liolaemus robustus and Liolaemus walkeri. We found interspecific differences in cell types in the chorion, allantois, and omphalopleure that may be explained by divergent phylogenetic history. Time elapsed since divergence may also explain the pronounced interspecific differences in vessel diameter, and within each species, there are strong differences in Dv between tissue locations. Both species show features to improve gas exchange in the chorioallantoic placenta including absence of eggshell, large Dv in the allantois (L. robustus) or embryonic side of the uterus (L. walkeri), and when present, microvillous cells in the allantois (L. walkeri). Both species also show features that suggest transfer of nutrients or water in the omphaloplacenta, including an almost complete reduction of the eggshell, secretive material (L. robustus), or vesicles (L. walkeri) on cell surface uterus, and when present specialized cells in the omphalopleure (L. walkeri). No statistical differences in Dv were found among stages 32–39 in each species, suggesting that a different mechanism, other than enhanced blood flow, might satisfy the increased oxygen demand of the developing embryos in the hypoxic environments of the high Andes. J. Morphol. 276:1205–1217, 2015. © 2015 Wiley Periodicals, Inc.     

Matrix effects break the LC behavior rule for analytes in LC-MS/MS analysis of biological samples

High-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are generally accepted as the preferred techniques for detecting and quantitating analytes of interest in biological matrices on the basis of the rule that one chemical compound yields one LC-peak with reliable retention time (Rt.). However, in the current study, we have found that under the same LC-MS conditions, the Rt. and shape of LC-peaks of bile acids in urine samples from animals fed dissimilar diets differed significantly among each other. To verify this matrix effect, 17 authentic bile acid standards were dissolved in pure methanol or in methanol containing extracts of urine from pigs consuming either breast milk or infant formula and analyzed by LC-MS/MS. The matrix components in urine from piglets fed formula significantly reduced the LC-peak Rt. and areas of bile acids. This is the first characterization of this matrix effect on Rt. in the literature. Moreover, the matrix effect resulted in an unexpected LC behavior: one single compound yielded two LC-peaks, which broke the rule of one LC-peak for one compound. The three bile acid standards which exhibited this unconventional LC behavior were chenodeoxycholic acid, deoxycholic acid, and glycocholic acid. One possible explanation for this effect is that some matrix components may have loosely bonded to analytes, which changed the time analytes were retained on a chromatography column and interfered with the ionization of analytes in the MS ion source to alter the peak area. This study indicates that a comprehensive understanding of matrix effects is needed towards improving the use of HPLC and LC-MS/MS techniques for qualitative and quantitative analyses of analytes in pharmacokinetics, proteomics/metabolomics, drug development, and sports drug testing, especially when LC-MS/MS data are analyzed by automation software where identification of an analyte is based on its exact molecular weight and Rt.