Synthesis and Incorporation of Pyrrole Carboxamide Nucleoside Triphosphates by DNA Polymerases

Abstract

We have synthesised and examined the enzymatic incorporation properties of the 5′-triphosphates of 2′-deoxyribosyl pyrrole 3-monocarboxamide (dMTP) and 2′-deoxyribosyl pyrrole 3,4-dicarboxamide (dDTP). These analogues we had hoped would behave as ambivalent base analogues in that they can present two alternative hydrogen-bonding faces either by rotation about the carboxamide group or about the glycosidic bond. The two pyrrole derivatives, dMTP and dDTP, exhibit a preference for incorporation with Klenow polymerase. They are preferentially incorporated as either A or C.     

Centrin homologues in higher plants are prominently associated with the developing cell plate

Summary Centrin and calmodulin are members of the EF-hand calcium-binding superfamily of proteins. In this study we compared localisation and immunoblotting of centrin with calmodulin in several monocot (onion and wheat) and dicot (mung bean andArabidopsis) plants. We confirmed that an anti-calmodulin antibody recognised a 17 kDa protein in all species tested and localises to the cytoplasm, mitotic matrix and with microtubules of the preprophase band and phragmoplast. In contrast, immunoblotting using anti-centrin antibodies shows that plant centrins vary from 17 to 20 kDa. Immunofluorescence microscopy with anti-centrin antibodies revealed only weak centrin immunoreactivity in the cytoplasm, nucleus, nuclear envelope, phragmoplast and mitotic matrix in meristematic cells. There was a slightly more intense perinuclear labelling in large differentiating onion cells and between separating anaphase chromosomes. While centrin is known to localise to the mitotic spindle poles in animal and algal cells, there was no appreciable immunoreactivity at the spindle poles in higher plants. In contrast, there was an intense immunofluorescence signal with anti-centrin antibodies in the developing cell plate. Further characterisation of the cell plate labelling by immunogold electron microscopy shows centrin immunoreactivity was closely associated with vesicles in the cell plate. Our observations suggest that centrin may play a role in cell plate formation.Abbreviations BSA bovine serum albumin - MTs microtubules - MTOCs microtubule organising centres - PBS phosphate buffered saline - PBST phosphate buffered saline with Tween-20     

根施甜菜碱对镍胁迫下小麦幼苗生长生理的影响

以普通小麦品种‘轮选988’为材料,采用溶液培养法,研究了根施不同浓度甜菜碱(1.0、2.0、3.0、4.0、5.0、10.0、15.0、20.0mmol·L~(-1))对镍(100μmol·L~(-1) NiSO_4)胁迫下小麦根系生长的影响,以及4.0mmol·L~(-1)甜菜碱处理镍胁迫幼苗根系相关抗逆生理生化指标的变化。结果表明:(1)与不施加镍对照相比,镍胁迫下小麦幼苗的根长、株高、鲜重和干重分别显著降低了14.7%、11.7%、15.0%和16.7%。(2)与单独镍胁迫处理相比,小麦幼苗的根长、株高、鲜重和干重均随着根施甜菜碱的浓度逐渐增加且呈先升后降的趋势,并以4.0mmol·L~(-1)外源甜菜碱处理效果较佳。(3)与单独镍胁迫处理相比较,在4.0mmol·L~(-1)外源甜菜碱处理下,小麦幼苗根系超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性分别升高了284.7%、40.3%、82.9%和20.4%,超氧阴离子自由基(O-·2)含量、过氧化氢(H_2O_2)含量和丙二醛(MDA)含量分别显著降低了50.6%、38.4%和40.6%,可溶性糖含量及游离脯氨酸(Pro)含量分别显著降低了19.2%、45.4%,而根系活力大幅上升了358.0%。研究认为,根施适宜浓度外源甜菜碱可显著增强小麦幼苗根系的抗氧化能力,恢复根系活力,从而有效减弱镍胁迫对小麦幼苗生长的伤害。     

Surface N balances and reactive N loss to the environment from global intensive agricultural production systems for the period 1970-2030

Data for the historical years 1970 and 1995 and the FAO-Agriculture Towards 2030 projection are used to calculate N inputs (N fertilizer, animal manure, biological N fixation and atmospheric deposition) and the N export from the field in harvested crops and grass and grass consumption by grazing animals. In most industrialized countries we see a gradual increase of the overall N recovery of the intensive agricultural production systems over the whole 1970-2030 period. In contrast, low N input systems in many developing countries sustained low crop yields for many years but at the cost of soil fertility by depleting soil nutrient pools. In most developing countries the N recovery will increase in the coming decades by increasing efficiencies of N use in both crop and livestock production systems. The surface balance surplus of N is lost from the agricultural system via different pathways, including NH3 volatilization, denitrification, N2O and NO emissions, and nitrate leaching from the root zone. Global NH3-N emissions from fertilizer and animal manure application and stored manure increased from 18 to 34 Tg·yr-1 between 1970 and 1995, and will further increase to 44 Tg·yr-1 in 2030. Similar developments are seen for N2O-N (2.0 Tg·yr-1 in 1970, 2.7 Tg·yr-1 in 1995 and 3.5 Tg·yr-1 in 2030) and NO-N emissions (1.1 Tg·yr-1 in 1970, 1.5Tg·yr-1 in 1995 and 2.0 Tg·yr-1 in 2030).     

Common Avian Infection Plagued the Tyrant Dinosaurs

Background

Tyrannosaurus rex and other tyrannosaurid fossils often display multiple, smooth-edged full-thickness erosive lesions on the mandible, either unilaterally or bilaterally. The cause of these lesions in the Tyrannosaurus rex specimen FMNH PR2081 (known informally by the name ‘Sue’) has previously been attributed to actinomycosis, a bacterial bone infection, or bite wounds from other tyrannosaurids.

Methodology/Principal Findings

We conducted an extensive survey of tyrannosaurid specimens and identified ten individuals with full-thickness erosive lesions. These lesions were described, measured and photographed for comparison with one another. We also conducted an extensive survey of related archosaurs for similar lesions. We show here that these lesions are consistent with those caused by an avian parasitic infection called trichomonosis, which causes similar abnormalities on the mandible of modern birds, in particular raptors.

Conclusions/Significance

This finding represents the first evidence for the ancient evolutionary origin of an avian transmissible disease in non-avian theropod dinosaurs. It also provides a valuable insight into the palaeobiology of these now extinct animals. Based on the frequency with which these lesions occur, we hypothesize that tyrannosaurids were commonly infected by a Trichomonas gallinae-like protozoan. For tyrannosaurid populations, the only non-avian dinosaur group that show trichomonosis-type lesions, it is likely that the disease became endemic and spread as a result of antagonistic intraspecific behavior, consumption of prey infected by a Trichomonas gallinae-like protozoan and possibly even cannibalism. The severity of trichomonosis-related lesions in specimens such as Tyrannosaurus rex FMNH PR2081 and Tyrannosaurus rex MOR 980, strongly suggests that these animals died as a direct result of this disease, mostly likely through starvation.     

The origin of modern crocodyliforms: new evidence from the Cretaceous of Australia

While the crocodyliform lineage extends back over 200 million years (Myr) to the Late Triassic, modern forms-members of Eusuchia-do not appear until the Cretaceous. Eusuchia includes the crown group Crocodylia, which comprises Crocodyloidea, Alligatoroidea and Gavialoidea. Fossils of non-crocodylian eusuchians are currently rare and, in most instances, fragmentary. Consequently, the transition from Neosuchia to Crocodylia has been one of the most poorly understood areas of crocodyliform evolution. Here we describe a new crocodyliform from the mid-Cretaceous (98-95 Myr ago; Albian-Cenomanian) Winton Formation of Queensland, Australia, as the most primitive member of Eusuchia. The anatomical changes associated with the emergence of this taxon indicate a pivotal shift in the feeding and locomotor behaviour of crocodyliforms-a shift that may be linked to the subsequent rapid diversification of Eusuchia 20 Myr later during the Late Cretaceous and Early Tertiary. While Laurasia (in particular North America) is the most likely ancestral area for Crocodylia, the biogeographic events associated with the origin of Eusuchia are more complex. Although the fossil evidence is limited, it now seems likely that at least part of the early history of Eusuchia transpired in Gondwana.     

Sensory nerve induced inflammation contributes to heterotopic ossification

Heterotopic ossification (HO), or bone formation in soft tissues, is often the result of traumatic injury. Much evidence has linked the release of BMPs (bone morphogenetic proteins) upon injury to this process. HO was once thought to be a rare occurrence, but recent statistics from the military suggest that as many as 60% of traumatic injuries, resulting from bomb blasts, have associated HO. In this study, we attempt to define the role of peripheral nerves in this process. Since BMP2 has been shown previously to induce release of the neuroinflammatory molecules, substance P (SP) and calcitonin gene related peptide (CGRP), from peripheral, sensory neurons, we examined this process in vivo. SP and CGRP are rapidly expressed upon delivery of BMP2 and remain elevated throughout bone formation. In animals lacking functional sensory neurons (TRPV1(-/-) ), BMP2-mediated increases in SP and CGRP were suppressed as compared to the normal animals, and HO was dramatically inhibited in these deficient mice, suggesting that neuroinflammation plays a functional role. Mast cells, known to be recruited by SP and CGRP, were elevated after BMP2 induction. These mast cells were localized to the nerve structures and underwent degranulation. When degranulation was inhibited using cromolyn, HO was again reduced significantly. Immunohistochemical analysis revealed nerves expressing the stem cell markers nanog and Klf4, as well as the osteoblast marker osterix, after BMP2 induction, in mice treated with cromolyn. The data collectively suggest that BMP2 can act directly on sensory neurons to induce neurogenic inflammation, resulting in nerve remodeling and the migration/release of osteogenic and other stem cells from the nerve. Further, blocking this process significantly reduces HO, suggesting that the stem cell population contributes to bone formation.