Studies on Anticodon-anticodon Interactions: Hemi-protonation of Cytosines Induces Self-pairing Through the GCC Anticodon of E. Coli tRNA-Gly

Abstract

The temperature-jump method was used to compare the stability of anticodon-anticodon duplexes formed by the self-association of two tRNAs: yeast tRNA-Asp and Escherichia coli tRNA-Gly. Yeast tRNA-Asp duplexes contain a U/U mismatch while E. coli tRNA-Gly dimers have a C/C mismatch in the middle position of their quasi self-complementary anticodons GUC and GCC, respectively. At neutral pH, it is found that only tRNA-Asp duplexes exist whereas at pH 5.0 only tRNA-Gly duplexes are formed. This reflects the hemi- protonation of the N3 of the cytosines at pH 5.0 which induces a pairing between the two middle residues of the anticodon GCC in E. coli tRNA-Gly. This is the first evidence that a protonated C-C(+) base pair is compatible with the formation of a double helix with antiparallel strands in a natural RNA molecule.     

阑尾杯状细胞类癌合并回盲部混合型腺神经内分泌癌1例病例报道并文献复习

目的:报道1例极为罕见的阑尾杯状细胞类癌(goblet cell carcinoid,GCC)合并回盲部混合型腺神经内分泌癌(mixed adenoneuroendocrine carcinoma,MANEC),以提高临床医师对本病的认识。方法:回顾性分析1例阑尾GCC合并回盲部MANEC患者的临床、病理特征、免疫组化及术后情况并进行文献复习。结果:该病例经病理切片会诊明确诊断为GCC,行右半结肠切除术后进一步诊断为回盲部MANEC,行1次FOLFOX化疗后,一般情况良好。结论:阑尾GCC合并回盲部MANEC是具有高度侵袭性的异质性恶性肿瘤,免疫组化局部Cg A和局部Syn阳性、Tang分类C组、临床分期IV期和非根治性手术为不良预后的危险因素。     

Identification of gonadotropic and antigonadotropic factors from the nervous part of the corpora cardiaca in the African locust

Summary

The nervous part of the locust corpora cardiaca (NCC) was extracted using 70% methanol and subjected to fast protein liquid chromatography (FPLC). Twenty fractions were collected and injected into females from the end of the IIIrd larval instar to day-12 of the adult life to screen the effects on metamorphosis, body pigmentation, and oocyte growth. The rate of juvenile hormone (JH) biosynthesis by the corpora allata of injected animals was evaluated in vitro. One fraction was found to stimulate ovarian maturation, and 3 to delay oocyte growth. None of them disturbed (i) either metamorphosis and body color (which are JH dependent), or (ii) rate of JH biosynthesis. Of the 3 fractions which inhibited ovarian maturation, 2 of them were identified as the neuroparsins A and B. The results clearly illustrate that antagonist factors from the brain are involved in the regulation of ovarian maturation in the locust.     

The activity of denervated corpora allata in a diapausing strain of Locusta migratoria: in vivo and in vitro studies

Summary

The activity of denervated corpora allata of a diapausing strain of Locusta migratoria is compared in vivo and in vitro. Inactive corpora allata (CA) of diapausing adult females soon become active following implantation into other diapausing females. Activation also follows incubation in vitro, although this activity decreases rapidly. Active CA of animals whose diapause has been broken by NCC2 section continue to secrete hormone when implanted into host animals (i.e., in vivo) although activity declines after 3 days. In contrast such CA show very low activity in vitro. These data confirm that in L. migratoria activity of the CA is regulated by a dual control involving inhibiting and activating factors whose origins are discussed.     

Separation and characteristics of antidiuretic factors from the corpora cardiaca of the migratory locust

Summary

Corpora cardiaca (CC) of the migratory locust contain two antidiuretic (AD) factors increasing the fluid reabsorption of isolated recta over a 5-h period.

They are contained in storage lobes (SL) and glandular lobes (GL) of the CC. They differ in size and can be separated from each other in different ways (extraction, dialysis, electrophoresis).

The AD factor contained in GL of the CC is stable at 4°C and quickly destroyed at 100°C. It stimulates the fluid reabsorption in a dose-dependent manner. Its release into the haemolymph seems to be controlled by octopamine, which is probably synthesized in the lateral neurosecretory areas of the brain.     

Synthesis, characterization, and evaluation of a novel 99mTc(CO)3 pyrazolyl conjugate of a peptide nucleic acid sequence

The 16-mer peptide nucleic acid sequence H-A GAT CAT GCC CGG CAT-Lys-NH₂ (1), which is complementary to the translation start region of the N-myc oncogene messenger RNA, was synthesized and conjugated to a pyrazolyl diamine bifunctional chelator (pz). The novel conjugate pz-A GAT CAT GCC CGG CAT-Lys-NH₂ (2) was labeled with technetium tricarbonyl, yielding quantitatively the complex fac-[^99mTc(CO)₃(κ³-pz-A GAT CAT GCC CGG CAT-Lys-NH₂)]²⁺ (4). Complex 4 was obtained with high radiochemical purity and high specific activity, revealing high stability in human serum and in cell culture medium. The identity of 4 was confirmed by comparing its reversed-phase high performance liquid chromatography profile with that of the rhenium analog fac-[Re(CO)₃(κ³-pz-A GAT CAT GCC CGG CAT-Lys-NH₂)]²⁺ (3), prepared by conjugation of fac-[Re(CO)₃(3,5-Me₂pz(CH₂)₂N((CH₂)₃COOH)(CH₂)₂NH₂)]⁺ to 1, using solid-phase techniques. UV melting experiments of 1 and 3 with the complementary DNA sequence led to the formation of stable duplexes, indicating that the conjugation of 1 to the pyrazolyl chelator and to the metal fragment fac-[M(CO)₃]⁺ did not affect the recognition of the complementary sequence as well as the duplex stability. For a first screening, SH-SY5Y human neuroblastoma cells, which express N-myc, were treated with 4. The results show that 4 internalizes (7% of the activity goes into the cells, after 4 h at 37 °C), presenting also a relatively high cellular retention (only 40% of internalized activity is released from the cells after 5 h). An erratum to this article can be found at     

Chlorophyll Catabolites in Senescent Leaves of the Lime Tree (Tilia cordata)

In cold extracts of senescent leaves of the Lime tree (Tilia cordata), two colorless nonfluorescent chlorophyll catabolites (NCCs) were identified, named Tc‐NCC‐1 and Tc‐NCC‐2, as well as a polar yellow chlorophyll catabolite (YCC), named Tc‐YCC. The constitution of the two NCCs was determined by spectroscopic means. In addition, a tentative structure was derived for Tc‐YCC. The three chlorophyll degradation products exhibited tetrapyrrolic structures, as are typical of NCCs or YCCs, and turned out to be rather polar, due to a glucopyranosyl group at their 8²‐position. At their 3‐positions, the more polar Tc‐NCC‐1 carried a 1,2‐dihydroxyethyl group and the less polar Tc‐NCC‐2 a vinyl group. Tc‐YCC was identified as the product of an oxidation of Tc‐NCC‐1.     

V1-Vasopressin Receptors in the Septum of the Rat Brain. Electrophysiological Evidence

Abstract

In slices from the rat brain, extracellular recordings were obtained from single neurones located in the lateral septum, an area known to receive a vasopressinergic innervation. Approximately half of the neurones tested responded to vasopressin by a concentration-dependent increase in firing rate, the lowest effective concentration being in the order of 2 nM. The effect of vasopressin was blocked by a synthetic structural analogue possessing vasopressor and oxytocic antagonistic properties on peripheral vasopressin and oxytocin receptors. Oxytocin had a weak effect in firing septal neurones, whereas a selective oxytocic agonist was totally ineffective. The action of vasopressin on neuronal firing was mimicked by a vasopressor agonist (Phe2-Orn8-VT) but not by a selective antidiuretic agonist (dDAVP). These results indicate that the vasopressin receptors present in rat septum are V1 (vasopressor type) rather than V2 (antidiuretic type) receptors. In addition, we conclude that these receptors, when occupied, lead to increased firing of lateral septal neurones.     

Exogenous retinoic acid decreases in vivo and in vitro proliferative activity during the early migratory stage of neural crest cells

We have previously demonstrated that directional migration of neural crest cells (NCC) is associated with a high cell density, resulting from an active cell proliferation. It is also known that treatment with retinoic acid (RA) causes a dose-dependent inhibition of proliferation of some cell types, and that administration of RA during the early stages of embryonic development, induces cranio-facial abnormal patterns corresponding to NCC derivatives. In view of these findings, it was of interest to determine if exogenous RA is a potential modulator of the mitotic rate of NCC, and to explore the hypothesis of an inhibitory effect exerted by RA on the proliferative behaviour of NCC in vivo and in vitro. Homogenates of RA-treated chick embryos showed a low [³H]dT incorporation, indicating a generalized diminution of DNA synthesis. The labelling index (LI=number of labelled cells/total number of cells) revealed that NCC from RA-treated and control embryos had higher values of [³H]dT incorporation than neural tube cells (P < 0.0001). Autoradiographs of RA-treated chick embryos showed a significantly lower [³H]dT incorporation in NCC at the prosencephalic and mesencephalic levels, as well as in the neural tube cells at the prosencephalic, mesencephalic and rhombencephalic levels, than in control chick embryos (P < 0.0001). NCC cultures treated with 1 or 10 μm RA had a significantly lower LI than in cultures treated with 0.1 μm RA or control cultures (P < 0.04). In chick embryos, the mitotic index of NCC was 0.026 for RA-treated and 0.033 for controls, while the duration of the cell cycle was significantly longer in the NCC of RA-treated embryos (～ 40 h) than in controls (～ 25 h). The length of the cell cycle phases of NCC was similar in both experimental conditions, except for G₁ phase, which was significantly longer in the RA-treated group than in controls. These results show that RA blocks DNA synthesis and lengthens the proliferative behaviour of NCC both in early chick embryos and in vitro, effects that could modify the morphogenetic patterns of NCC distribution through a decreased cell population.     

Patterns of phylogenetic community structure of sand dune plant communities in the Yucatan Peninsula: the role of deterministic and stochastic processes in community assembly

ABSTRACT

Background: Tropical sand dunes are ideal systems for understanding drivers of community assembly as dunes are subject to both deterministic and stochastic processes. However, studies that evaluate the factors that mediate plant community assembly in these ecosystems are few.

Aims: We evaluated phylogenetic community structure to elucidate the role of deterministic and stochastic processes in mediating the assembly of plant communities along the north of the Yucatan Peninsula, Mexico.

Methods: We used plastid genetic markers to evaluate phylogenetic relationships in 16 sand-dune communities. To evaluate the role of climate in shaping plant community structure we carried out linear regressions between climatic variables and mean phylogenetic distance. We estimated the Net Relatedness Index and Nearest Taxon Index to identify ecological processes mediating community assembly.

Results: Observed phylogenetic structure was not different from random, suggesting that stochastic processes are the major determinants of community assembly. Climate was slightly correlated with phylogenetic diversity suggesting that abiotic environment plays a minimal role in community assembly.

Conclusions: Random assembly appears to be the primary factor structuring the studied sand dune plant communities. Environmental filters may represent a secondary factor contributing to the observed phylogenetic structure. Thus, both processes may act simultaneously to mediate the assembly of sand-dune plant communities.     

Porcine model of neurocysticercosis by intracarotid injection of Taenia solium oncospheres: Dose assessment,infection outcomes and serological responses

BackgroundNeurocysticercosis (NCC) is the infection of the human central nervous system (CNS) by Taenia solium larvae that cause significant neurological morbidity. Studies on NCC pathophysiology, host-parasite interactions or therapeutic agents are limited by the lack of suitable animal models. We have previously reported that carotid injection of activated T. solium oncospheres directs parasites into the CNS and consistently reproduces NCC. This study assessed the minimal dose required to consistently obtain NCC by intracarotid oncosphere injection and compared antigen and antibody response profiles by dose-group.Methods/Principal findingsThree groups of pigs were infected with either 2500 (n = 10), 5000 (n = 11), or 10000 (n = 10) oncospheres. Two pigs died during the study. Necropsy exam at day 150 post-infection (PI) demonstrated viable NCC in 21/29 pigs (72.4%), with higher NCC rates with increasing oncosphere doses (4/9 [44.4%], 9/11 [81.8%] and 8/9 [88.9%] for 2500, 5000, and 10000 oncospheres respectively, P for trend = 0.035). CNS cyst burden was also higher in pigs with increasing doses (P for trend = 0.008). Viable and degenerated muscle cysticerci were also found in all pigs, with degenerated cysticerci more frequent in the 2500 oncosphere dose-group. All pigs were positive for circulating parasite antigens on ELISA (Ag-ELISA) from day 14 PI; circulating antigens markedly increased at day 30 PI and remained high with plateau levels in pigs infected with either 5000 or 10000 oncospheres, but not in pigs infected with 2500 oncospheres. Specific antibodies appeared at day 30 PI and were not different between dose-groups.Conclusion/SignificanceIntracarotid injection of 5000 or more oncospheres produces high NCC rates in pigs with CNS cyst burdens like those usually found in human NCC, making this model appropriate for studies on the pathogenesis of NCC and the effects of antiparasitic treatment.     

Report of the Annual Conference of the Cryptogamic Section, 1948

Background: Trait-based assembly rules are a powerful tool in community ecology, used to explore the pattern and process of community structure (richness and composition).

Aims: A preliminary test for the utility of trait-based assembly rules in explaining cryptogamic epiphyte communities (lichens and bryophytes).

Methods: We sampled epiphytes from three different tree species (aspen, birch and pine), and from trees of contrasting age. The community composition of epiphyte species (taxon analysis) and functional groups (trait analysis) was summarised using multivariate ordination (nonmetric multidimensional scaling, NMDS).

Results: Ordination documented a widely observed pattern in which different tree species have taxonomically different epiphyte communities. However, NMDS sample scores were correlated to tree age in the trait-based analysis, but not in the taxon analysis.

Conclusions: Our results point to the existence of a common pattern in community traits during succession (on trees of different age) when measured for epiphyte communities with contrasting taxonomic composition. This pattern is evidenced by consistent trends in lichen growth form and reproductive strategy (sexual vs. asexual).     

Prevalence and clinicopathologic characteristics of gastric cardia cancer in South Korea

Background and Aim: Western reports have suggested that the prevalence of gastric cardia cancer (GCC) has been increasing, and indicated some differences between GCC and gastric noncardia cancer (GNCC). However, few studies have been conducted in Asia. The aims of this study were to estimate the prevalence of GCC and to evaluate differences of clinicopathologic characteristics between GCC and GNCC in South Korea. Methods: This study was single‐center case–control study. A total of 829 patients with gastric cancer and 270 controls were enrolled between 2003 and 2011. Baseline characteristics, Helicobacter pylori (H. pylori) infection status, and histologic characteristics were compared among three groups (GCC, GNCC, and control). Results: Sixty cases (7.2%) of gastric cancer were located in cardia. Multivariate analysis showed that male odds ratio (OR, 5.72; 95% CI, 1.72–19.07; p = .005) and cigarette smoking (OR, 5.38; 95% CI, 1.39–20.90; p = .015) were risk factors of GCC in comparison with control group, but H. pylori infection rate was not significant. In the case of GNCC, cigarette smoking (OR, 3.87; 95% CI, 1.81–8.29; p < .001), past alcohol intake (OR, 2.82; 95% CI, 1.28–6.20; p = .010), intestinal metaplasia (OR, 3.22; 95% CI, 2.00–5.17; p < .001), and H. pylori infection (OR, 3.06; 95% CI, 1.90–4.93; p < .001) were risk factors of GNCC. Gastroesophageal reflux disease symptoms were higher in the GNCC (21.2%) than control group (13.5%) (p = .008). However, in the case of GCC, they were similar between the GCC (12.7%) and control group (p = .872). According to multivariate analysis, history of H. pylori eradication (OR, 0.34; 95% CI, 0.19–0.61; p < .001) was associated with a protective effect on GNCC. GCC showed higher depth of invasion (p = .038) and frequent distant metastasis (p = .012) than GNCC. Conclusion: In this referral center based study, the prevalence of GCC was 7.2% in South Korea. Risk factors and clinicopathologic characteristics for GCC and GNCC were different, supporting that the pathophysiology is different in the development of GCC and GNCC.     

Cinnamic,myristic and fumaric acids in tobacco root exudates induce the infection of plants by Ralstonia solanacearum

Aim

The secretion of allelochemicals from plant roots plays a key role in soil sickness and soil-borne disease. The goal of this study was to investigate the role of allelopathic chemicals in Ralstonia solanacearum-infected tobacco roots.

Methods

The organic acids investigated in the present study are major components of tobacco root exudates. Through a swarming assay, we assessed the chemotaxis and colonization of R. solanacearum in response to organic acids.

Results

Fumaric acid was detected, and the results showed that this acid could serve as a semiochemical for attracting R. solanacearum and inducing the formation of biofilms of this species. The results also revealed that cinnamic and myristic acids play significant roles on swarming motility and chemotaxis. In addition, cinnamic, myristic and fumaric acids could enhance the expression of chemotaxis- and motility-related genes in R. solanacearum cultured in minimal medium. Furthermore, these three acids promote R. solanacearum colonization and accelerate disease progression in tobacco.

Conclusion

Cinnamic, myristic and fumaric acids could serve as semiochemical attractants to induce the colonization and infection of R. solanacearum. The results of the present study enhance our understanding of the ecological effects of plant root exudates in plant-microbe interactions and help to reveal the relationship between tobacco bacterial wilt and the autotoxins and allelochemicals that accumulate from root exudates.

     

Cholinesterase inhibitory triterpenoids from the bark of Garcinia hombroniana

Abstract

Context: Garcinia hombroniana Pierre, known as manggis hutan in Malaysia is a rich source of xanthones and benzophenones.

Objectives: This study was aimed to isolate and characterize potential cholinesterase inhibitors from the extracts of G. hombroniana bark and investigate their interactions with the enzymes.

Materials and methods: The dichloromethane extract afforded five triterpenoids which were characterized by NMR and mass spectral techniques. Cholinesterase inhibitory assay and molecular docking were performed to get insight of the inhibitory activity and molecular interactions of the compounds. The compounds were also tested for their antioxidant capacity.

Results: The isolated triterpenoids were identified as: 2β-hydroxy-3α-O-caffeoyltaraxar-14-en-28-oic acid (1), taraxerol (2), taraxerone (3), betulin (4) and betulinic acid (5). Compound 1 was the most active dual inhibitor of both AChE and BChE. Compound 1 also showed good antioxidant activities.

Conclusion: Compound 1 had dual and moderate inhibitory activity on AChE and BChE worthy for further investigations.     

Numerical analysis of phenotypic properties,genomic fingerprinting,and multilocus sequence analysis of Bradyrhizobium strains isolated from root nodules of Lembotropis nigricans of the tribe Genisteae

Purpose

The aim of this study was to estimate the level of genomic and phenotypic diversity as well as the genus and species position of bacterial strains isolated from root nodules of Lembotropis nigricans (family Fabaceae).

Methods

The genomic diversity of studied L. nigricans nodule symbionts was examined by using BOX-PCR and AFLP (amplified fragment length polymorphism) fingerprinting techniques. To assign bacteria to the genus, numerical analysis of phenotypic features and comparative analysis of 16S rDNA sequences were performed. The comparative analysis of combined atpD, dnaK, gyrB, and rpoB gene sequences (multilocus sequence analysis, MLSA) was used to determine the most closely related species to the studied bacteria.

Results

Both BOX-PCR and AFLP techniques revealed a high level of genomic heterogeneity of L. nigricans nodulators. Among 33 studied bacteria, 32 genotypes were delineated by the AFLP method and 27 genotypes were identified by the BOX-PCR fingerprinting. The numerical analysis of 86 phenotypic characteristics of L. nigricans nodule isolates and reference rhizobia showed that studied bacteria belong to the genus Bradyrhizobium. Affiliation of L. nigricans nodule isolates to the genus Bradyrhizobium was supported by comparative analysis of 16S rDNA sequences and the concatenation of atpD, dnaK, gyrB, and rpoB gene sequences. MLSA indicated also that L. nigricans microsymbionts are members of Bradyrhizobium japonicum.

Conclusion

L. nigricans root nodule symbionts are members of Bradyrhizobium japonicum and exhibit high phenotypic and genomic diversity important for their survival in soil.

     

On the Nature of Isomeric Nonfluorescent Chlorophyll Catabolites in Leaves and Fruit – A Study with a Ubiquitous Phylloleucobilin and its Main Isomerization Product

The typical main products of chlorophyll (Chl) breakdown in higher plants are non‐fluorescent, colorless phyllobilins, named phylloleucobilins. These long elusive Chl‐catabolites are linear tetrapyrroles, whose structure elucidation has required thorough spectroscopic analyses. Interestingly, in recent LC/MS studies of leaf extracts, isomeric forms of phylloleucobilins were detected. The existence of isomeric phyllobilins may suggest incomplete stereo‐selectivity of catabolic processes, or isomerization processes in plant cells or in the analytes. Here we report a study with the phylloleucobilin NCC‐1, a basic Chl‐catabolite in extracts of leaves and fruit. NCC‐1 and its main isomerization product in aqueous solution were identified as 8²‐epimers. Formation of 8²‐epi‐NCC‐1 from NCC‐1 implies an unstable enol(ate)‐intermediate, which reverts to NCC‐1 or converts to 8²‐epi‐NCC‐1. Such reversible epimerization reactions are a non‐biological in vitro feature of typical phylloleucobilins, and probably also take place in vivo.     

The projection of neuroendocrine fibers (NCC I and II) in the brains of three orthopteroid insects

Neuronal projections from neuroendocrine tracts (nervi corpori cordiaci I and II) in the brains of the locust (Schistocerca vaga), cricket (Acheta domesticus), and cockroach (Periplaneta americana) were studied using reconstructions of silver-intensified cobalt chloride preparations. Collaterals from the NCC I in these species branch extensively in the dorsal protocerebral neuropile, anterior to the stalk of the corpora pedunculata and ventral to its calyces. Other fibers project from the NCC I bilaterally into the medial protocerebral neuropile, anterior to the central body, and posterior to the beta lobes. NCC II collaterals arborize in the medial, dorsal, and lateral protocerebral neuropile, their region of projection partially overlapping with that of the NCC I. Several NCC II fibers terminate in the superior arch of the central body in Acheta but not in the other two species. Tritocerebral cells filled through the NCC I branch in the medial tritocerebral neuropile in all three species, but most extensively in Schistocerca. No NCC fibers were seen to penetrate any part of the corpora pedunculata, protocerebral bridge, olfactory glomeruli, ocellar tracts, or optic lobes. These neuronal projections from the NCC I and II lie anterior to regions of branching of second-order ocellar fibers and thus provide no anatomical basis for direct ocellar input to neurosecretory cells, contrary to previous reports for orthopteroid species (Brousse-Gaury, '71a, b). However, interneurons filled from the optic lobes were found to terminate in the same region of dorsal protocerebral neuropile as NCC I and II fibers in Acheta, thus providing a possible pathway for optic input to the cerebral neuroendocrine system.     

Three New Species of Gelechiid Moths (Lepidoptera,Gelechiidae) from the Southeastern Regions of the European Part of Russia

Three new species of gelechiid moths (Gelechiidae) are described from the arid regions of the southeastern European part of Russia (Astrakhan and Orenburg provinces): Bryotropha satschkovisp. n., Monochroa aenigmasp. n., and Scrobipalpa achtubicasp. n.

     

Investigation of the relationship between kinesiophobia and sensory processing in fibromyalgia patients

Abstract

Purpose/aim: The aims of this study are to investigate the relationship between kinesiophobia and sensory processing in fibromyalgia (FM) patients and obtain new information about kinesiophobia in light of these results.

Materials and methods: This study has been conducted with 82 literate subjects aged 18–65?years and diagnosed with FM based on ACR 2010 diagnostic criteria. Fibromyalgia Impact Questionnaire was used for measuring functional status in FM patients, The Tampa Scale of Kinesiophobia was used for determine the levels of kinesiophobia and Adolescent/Adult Sensory Profile was used for determine the characteristics of sensory process patterns of the individuals.

Results: Among the participants, sensory sensitivity scores of 65.85% and sensation avoiding scores of 40.24% were higher than that in the general population. Sensation seeking scores of 48.78% the subjects were lower compared to the general population. A significant and weak positive correlation was found between the kinesiophobia scores and responses of sensory sensitivity and sensation avoiding (r?=?0.23, p?=?0.04; z?=?0.29, p?=?0.01)

Conclusion: This is the first study conducted to investigate the relationship between kinesiophobia and sensory processing in fibromyalgia patients. However, different studies investigating this subject are warranted in order to be able to generalize the findings and increase the value of evidence.