The specificity of the syngeneic mixed leukocyte response, a primary anti-I region T cell proliferative response, is determined intrathymically

In previous studies, the syngeneic MLR of peripheral T cells was shown to be predominantly an I region-restricted function. In this report we show that adult thymocytes are also capable of responding to syngeneic irradiated stimulator cells in a syngeneic MLR, provided that TCGF is added to the culture system. Using this assay, it was possible for the first time to examine the pattern of I region restriction within the thymus itself. Analysis of the thymocyte syngeneic MLR in thymuses from radiation-induced bone marrow chimeras demonstrated that the MHC preference seen in the peripheral T cell population also existed in cells resident within the thymus. Experiments utilizing congenitally athymic mice transplanted with allogeneic thymic grafts demonstrated that both peripheral T cells and thymocytes from such animals displayed a strong preferential proliferation toward stimulator cells bearing thymic-type MHC determinants. The results in the nude model thus demonstrate that the thymus by itself is sufficient to impart such restriction specificity on a developing T cell repertoire. These results are consistent with the notion that the thymus exerts selective pressure on maturing T cell populations that results in a skewing of the T cell repertoire toward the recognition of thymic-type I region products, and that this MHC preference exists before expansion of T cells in the periphery.     

Dietary restriction causes chronic elevation of corticosterone and enhances stress response in red-legged kittiwake chicks

Release of corticosterone in hungry kittiwake chicks facilitates begging and allows them to restore depleted energy reserves by increasing parental food provisioning. However, in order to avoid detrimental effects of chronic elevation of corticosterone, chicks might suppress adrenocortical activity in response to prolonged food shortages. In this study we examined temporal dynamics of corticosterone release in red-legged kittiwake (Rissa brevirostris) chicks exposed to prolonged restrictions in energy content and/or nutritional quality (low versus high lipid content) of their food. Starting at the age of 15 days, chicks were fed either high- or low-lipid fish at 40%, 65%, and 100% of ad libitum energy intake. Body mass measurements and baseline plasma samples were taken on a weekly basis after beginning of the treatment. After 3 weeks of treatment, chicks were exposed to a standardized acute handling and restraint stress protocol, where in addition to a baseline sample, three plasma samples were taken at intervals up to 50 min. We found that food-restricted chicks had lower body mass, chronically (during 2-3 weeks) elevated baseline and higher acute stress-induced levels of corticosterone compared to chicks fed ad libitum. Low lipid content of food further exacerbated these effects. An increase in baseline levels of corticosterone was observed within a week after energy requirements of food-restricted chicks exceeded their daily energy intake. A tendency for suppression of adrenocortical activity was observed in treatments fed low-lipid diets only at the end of the experiment. We suggest that nest-bound chicks, if food-stressed, might suffer deleterious effects of chronic elevation of corticosterone.     

Potential new areas for conservation of key botanical families in the subtropical Atlantic Forest

Myrtaceae, Lauraceae and Fabaceae are regarded as essential floristic elements of Atlantic forests due to their outstanding species richness, endemism levels, and ecological functions. Nonetheless, Atlantic forests are being subjected to multiple human disturbances that compromise the conservation of their flora. This study, therefore, intended to address whether there exist potential areas for conservation with great richness of tree/shrub species of the aforementioned families in different forest types within the subtropical Atlantic Forest. For this, data collected systematically across?~?23% of the subtropical Brazilian Atlantic Forest were employed. The univariate Local Moran I statistic was used to search for clusters of sample plots with great richness of species of the studied families. Six clusters were found throughout the evergreen rainforest (ERF) and Araucaria forest (AF), and most of them contained more than half of all species of these families observed on the sample plots, besides many others belonging to different families. A cluster of Myrtaceae and a cluster of Lauraceae in the ERF were the only ones that overlapped protected areas. The clusters of Lauraceae in the AF, located in ecotone zones with the ERF, had?~?50% of native forest cover, whereas the clusters of Myrtaceae and Fabaceae had?~?10% of forest cover. Inasmuch as forests in the study area have been heavily exploited, the clusters have relevant conservation value. Protected areas could be expanded or converted into more restrictive conservation categories to enhance the conservation of populations of key elements of the Atlantic Forest. Yet, non-protected areas deserve attention regarding the management of forest resources and conservation-by-use strategies.

     

N-2-acetylaminofluorene modification of poly(dG-m5dC)·poly(dG-m5dC) induces the Z-DNA conformation

Poly(dG-m⁵dC)·poly(dG-m⁵dC) was modified by treatment with N-acetoxy-N-2-acetylaminofluorene (N-Aco-AAF) and its conformation examined by circular dichroism (CD) and susceptibility to S₁ nuclease digestion. A sample with a modification level of 10% shows a CD spectrum characteristic of the Z form and is resistant to digestion by S₁ nuclease. The relative reactivity of several polymers with N-Aco-AAF was shown to follow the order of ease of formation of Z DNA: poly(dG-m⁵dC)·poly(dG-m⁵dC) > poly(dG-dC)·poly(dG-dC) > poly(dG)·poly(dC). This suggests that AAF reacts more readily with Z DNA than B DNA.     

The binding of pentaammineruthenium (III) to RNase B and RNase A+d(pA)4 in the crystalline state

The binding of pentaammineruthenium (III) to ribonuclease A and B both free and complexed with d(pA)₄ has been examined in the crystalline state through the application of X-ray diffraction and difference Fourier techniques. In crystals of native RNase B, the reagent was observed to have many binding sites, some entirely electrostatic in nature and others consistent with coordination to histidine residues. The primary histidine in the latter case was 105 with 119 also partially substituted. In crystals of RNase A+d(pA)₄ complex only a single, extremely strong site of substitution was observed, and this was 2.4 Å from the native position of the imidazole ring of histidine 105. Thus, the results of these X-ray diffraction studies appear to be quite consistent with the findings of earlier NMR studies and with the results obtained in crystals of the gene 5 DNA binding protein.     

Evolutionary trends in Mio-Pliocene Leporinae, based onTrischizolagus (Mammalia, Lagomorpha)

New material ofTrischizolagus dumitrescuae from Moldova and Ukraine is described. The variation of p3 inTrischizolagus shows the gradual shift of morphotype frequencies from the ‘Hypolagus’ pattern in Turolian through the mixture of three patterns (including ‘Nekrolagus’ morphotype) in Early Ruscinian to the dominant ‘Alilepus’ pattern in the Late Ruscinian samples. These transformations took place parallel to that of the North AmericanNekrolagus. Probably North AmericanSylvilagus, Brachylagus, andRomerolagus had an North American origin fromNekrolagus, whereas Eurasiatic and AfricanOryctolagus, Caprolagus, Nesolagus, andPoelagus could have originated in the Old World fromTrischizolagus.     

Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints

We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP further enables simultaneous cloning of multiple inserts.