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Ribosome-inactivating proteins (RNA N-glycosidases) from the seeds of Saponaria ocymoides and Vaccaria pyramidata.

From the seeds of the Caryophyllaceae Saponaria ocymoides and Vaccaria pyramidata two proteins were purified which have the properties of the type-1 (single-chain) ribosome-inactivating proteins [reviewed by Barbieri, L., Battelli, M. G. & Stirpe, F. (1993) Ribosome-inactivating proteins from plants, Biochim. Biophys. Acta 1154, 237-282]. The proteins have molecular masses of 30.2 kDa (S. ocymoides) and 28.0 kDa (V. pyramidata) and pI greater than 9.5, their N-terminal amino acid sequences are similar to those of saporin-S6 and dianthin 30, ribosome-inactivating proteins from other Caryophyllaceae, and they partially cross-react with sera against these proteins. Both proteins inhibit protein synthesis by a rabbit-reticulocyte lysate with IC50 (concentrations giving 50% inhibition) below 10(-10) M, have a smaller effect on poly(U)-directed phenylalanine polymerisation by rat liver ribosomes (nanomolar IC50, approximately) and on protein synthesis by various cell lines (IC50 ranging from 4 nM to > 3000 nM) and possess rRNA N-glycosidase activity, releasing 1 mol adenine/ribosome.

Amino Acid Sequence↗

Thionation of segetalins A and B, cyclic peptides with estrogen-like activity from seeds of Vaccaria segetalis.

Thionation of estrogen-like active cyclic peptides, segetalins A (1) and B (2), with Lawesson's reagent provided each two thiosegetalins; thiosegetalin A1 [Gly-1-psi(CS-NH)-Val-2; Trp-5-psi (CS-NH)-Ala-6]segetalin A, thiosegetalin A2 [Gly-1-psi(CS-NH)-Val-2; Ala-6-psi(CS-NH)-Gly-1]segetalin A, thiosegetalin B1 [Gly-1-psi (CS-NH)-Val-2; Ala-3-psi(CS-NH)-Trp-4]segetalin B, and thiosegetalin B2 [Gly-1-psi(CS-NH)-Val-2; Trp-4-psi(CS-NH)-Ala-1]segetalin B. Thiosegetalin A2 only showed estrogen-like activity against ovariectomized rats. On the basis of their conformations analysed by NMR experiments, the backbone conformation was considered to play an important role in estrogen-like activity for segetalins.

Animals↗

Structural and functional analysis of Aplysia attractins, a family of water-borne protein pheromones with interspecific attractiveness.

Mate attraction in Aplysia involves a long-distance water-borne signal (the protein pheromone attractin), which is released during egg laying. Aplysia californica attractin attracts species that produce closely related attractins, such as Aplysia brasiliana, whose geographic distribution does not overlap that of A. californica. This finding suggests that other mollusks release attractin-related pheromones to form and maintain breeding aggregations. We describe four additional members of the attractin family: A. brasiliana, Aplysia fasciata, Aplysia depilans (which aggregates with A. fasciata aggregations), and Aplysia vaccaria (which aggregates with A. californica aggregations). On the basis of their sequence similarity with A. californica attractin, the attractin proteins fall into two groups: A. californica, A. brasiliana, and A. fasciata (91-95% identity), and A. depilans and A. vaccaria (41-43% identity). The sequence similarity within the attractin family, the conserved six cysteines, and the compact fold of the NMR solution structure of A. californica attractin suggest a common fold for this pheromone family containing two antiparallel helices. The second helix contains the IEECKTS sequence conserved in Aplysia attractins. Mutating surface-exposed charged residues within this heptapeptide sequence abolishes attractin activity, suggesting that the second helix is an essential part of the receptor-binding interface.

Amino Acid Sequence↗

Egg-laying hormone peptides in the aplysiidae family.

The neuropeptidergic bag cells of the marine mollusc Aplysia californica are involved in the egg-laying behavior of the animal. These neurosecretory cells synthesize an egg-laying hormone (ELH) precursor protein, yielding multiple bioactive peptides, including ELH, several bag cell peptides (BCP) and acidic peptide (AP). While immunohistochemical studies have involved a number of species, homologous peptides have been biochemically characterized in relatively few Aplysiidae species. In this study, a combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS) and electrospray ionization Fourier transform ion cyclotron resonance MS is used to characterize and compare the ELH peptides from related opisthobranch molluscs including Aplysia vaccaria and Phyllaplysia taylori. The peptide profiles of bag cells from these two Aplysiidae species are similar to that of A. californica bag cells. In an effort to characterize further several of these peptides, peptides from multiple groups of cells of each species were extracted, and microbore liquid chromatography was used to separate and isolate them. Several MS-based sequencing approaches are applied to obtain the primary structures of bag cell peptides and ELH. Our studies reveal that (&agr;)-BCPs are 100 % conserved across all species studied. In addition, the complete sequences of (&egr;)-BCP and ELH of A. vaccaria were determined. They show a high degree of homology to their counterparts in A. californica, with only a few amino acid residue substitutions.

Amino Acid Sequence↗

An immunohistochemical study of the neuropeptidergic bag cells of Aplysia.

The bag cell neurosecretory system of Aplysia californica synthesizes and releases a neuropeptide, the egg-laying hormone (ELH), which can induce egg-laying behavior. While ELH acts as a neurohormone on distant neuronal and non-neuronal targets, it also can moderate the electrical activity of neurons within the abdominal ganglion. Using antibodies generated against pure ELH, our immunohistochemical study provides direct morphological support for this dual function of ELH by defining the distribution of this neuropeptide within the abdominal ganglion of A. californica. We also find ELH cross-reactivity in the bag cell systems of three other species of Aplysia. In sections of abdominal ganglia stained for ELH by the peroxidase-antiperoxidase method, bag cell clusters were found to be homogeneous populations of ELH-immunoreactive neurons, extending processes in a complex array within the connective tissue sheath overlying the ganglion and the proximal regions of neighboring nerves. This association of neurites with the sheath is the neurohemal organ which facilitates release of ELH into circulating hemolymph. Stained processes also invade the interior of the ganglion in association with septal sheets of connective tissue and as scattered fibers in the neuropil. Local modulation by ELH probably occurs through release from these intraganglionic sites. A few ELH-positive ectopic cells were seen outside of the confines of the bag cell clusters as far displaced as the base of the branchial nerve; they appear to be bag cells which have mis-migrated during development. Specificity of the antiserum was confirmed by the total lack of staining of all other neurons within the abdominal ganglion as well as the absence of staining of bag cells with pre-immune serum. Although th ELHs of Aplysia vaccaria, Aplysia braziliana, and Aplysia dactylomela have not been characterized biochemically, it is known that abdominal ganglion extracts from A. californica can induce egg laying in all three species. We find that antiserum against A. californica ELH stains the bag cell systems of A. vaccaria, A. braziliana, and A. dactylomela and not other parts of their abdominal ganglia. It is likely that this staining represents cross-immunoreactivity of the egg-laying neurohormones from these species which also share cross-bioactivity.

Animals↗

Comparative efficacy of five herbicides on winter cereal weeds in semi-arid region of Algeria.

Field chemical control trials on cereal weeds were conducted in the Sétif region (north-east of Algeria), using five herbicides. The most abundant weeds were Fumaria densiflora DC., Veronica cymbalaria Bodard, Vaccaria pyramidata Medik, and Papaver rhoeas L.. The herbicides applied at the end-tillering stage of wheat were bromoxynil + diclofop-methyl, flamprop-isopropyl + MCPA, 2,4-D ester, diclofop-methyl and tribenuron-methyl. Compared to the untreated control, they increased grain yield by 359, 294, 216, 174 and 417 kg ha-1, respectively. These results showed the importance of the chemical control, which is applied in the Sétif region on less than 5% of the total surface of cereals.

Algeria↗

In vitro anticancer activity of twelve Chinese medicinal herbs.

Aqueous extracts of 12 Chinese medicinal herbs, Anemarrhena asphodeloides, Artemisia argyi, Commiphora myrrha, Duchesnea indica, Gleditsia sinensis, Ligustrum lucidum, Rheum palmatum, Rubia cordifolia, Salvia chinensis, Scutellaria barbata, Uncaria rhychophylla and Vaccaria segetalis were evaluated for their antiproliferative activity on eight cancer cell lines as well as on normal human mammary epithelial cells. Five human and three murine cancer cell lines representing different tissues (breast, lung, pancreas and prostate) were used. All the crude aqueous extracts demonstrated growth inhibitory activity on some or all of the cancer cell lines, but only two showed activity against the normal mammary epithelial cells. Overall, the murine cell lines tended to be more sensitive to most of the extracts compared with the human cell lines. Among the human cell lines, cell type specificity was observed for two extracts. These results indicate the potential use of traditional Chinese medicinal herbs as antineoplastic agents and suggest that further studies evaluating their mechanism(s) of action and the isolation of active antitumor compounds are warranted.

Animals↗

Auriculotherapy on low back pain in the elderly.

The objective of the study was to examine the effectiveness of auriculotherapy using magnetic pellets for the elderly suffering from low back pain (LBP). Sixty participants who were 60-years old or above and had been suffering from LBP were recruited. Participants were randomly allocated to receive auriculotherapy on a 3-week basis using either Semen Vaccariae (control group=30) or magnetic pellets (experimental group=30). Seven auricular acupoints that are believed to have an effect on LBP were selected. Treatment effects were evaluated using the Chinese Pain Intensity Verbal Rating scale (VRS). The experimental group had indeed experienced a significant improvement in pain relief when compared with the control group; and the therapeutic effects were sustained at 2 and 4-week follow-up periods after the therapy. Findings of this study demonstrated that auriculotherapy using magnetic pellets significantly reduce the pain intensity level of the elderly suffering from non-specific LBP.

Acupressure↗

A conserved heptapeptide sequence in the waterborne attractin pheromone stimulates mate attraction in Aplysia.

Mate attraction in the marine mollusk Aplysia involves long-distance waterborne chemical signaling via the release of the peptide pheromone attractin during egg laying. Aplysia californica attractin attracts conspecifics, reduces the latency to mating, and stimulates hermaphroditic mating. Four additional members of the Aplysia attractin family have recently been characterized from Aplysia brasiliana, Aplysia fasciata, Aplysia depilans, and Aplysia vaccaria. The five sequences differ significantly, but share six cysteine residues and the strictly conserved sequence Ile30-Glu-Glu-Cys-Lys-Thr-Ser36. Attractin is attractive to geographically and evolutionarily distant species, suggesting that the conserved heptapeptide region may be important for mate attraction. Consistent with this prediction, a synthetic constrained cyclic peptide that contains the conserved heptapeptide sequence is significantly attractive in T-maze bioassays. The attractins are the first family of waterborne peptide pheromones characterized in invertebrates and are unique in that family members are not species-specific pheromonal attractants.

Amino Acid Sequence↗

Molluscan attractins, a family of water-borne protein pheromones with interspecific attractiveness.

The marine mollusk Aplysia releases the water-borne pheromone attractin during egg laying. This small protein stimulates the formation and maintenance of mating and egg-laying aggregations. Attractin has been characterized from five Aplysia species: A. californica, A. brasiliana, A. fasciata, A. vaccaria, and A. depilans. We describe here the isolation of attractin from Bursatella leachii, and show that it belongs to the same protein family. The pattern of residue conservation, especially the six invariant cysteines, suggests that all of these attractins have a common fold. The nuclear magnetic resonance solution structure of A. californica attractin contains two antiparallel alpha-helices, the second of which contains the heptapeptide sequence IEECKTS that has been implicated in attractin function. Synthetic peptides containing this IEECKTS region are attractive, and mutating surface exposed charged residues within this region of attractin abolishes attractin activity. This suggests that the second helix is an essential part of the receptor-binding interface. In contrast to the peptide pheromonal attractants in amphibians, which are species specific, the attractins are, to our knowledge, the first water-borne peptide or protein pheromone family in invertebrates and vertebrates that are not species specific.

Amino Acid Sequence↗

Effectiveness of auricular therapy on sleep promotion in the elderly.

Sleep disturbances are a particularly common problem in the elderly. The purpose of this study was to examine the effectiveness of auricular therapy on sleep behaviors in the elderly. One hundred and twenty participants of 60 years old or above and who were suffering from sleep disturbances were invited to participate in this study. Eligible participants were randomly allocated to receive auricular therapy using Junci Medulla (Group A = 30), Semen Vaccariae (Group B = 30) or magnetic pearls (Group C = 60). Groups A and B were the control groups, while Group C was the experimental group. Seven auricular points which are thought to have an effect on promoting sleep were selected. The total treatment course lasted for three weeks. Objective measurement using actigraphic monitoring was performed before the therapy commenced, in the middle period of the therapy, and within one week after the therapy had been completed. After the therapy, there were significant differences among the three groups in terms of the nocturnal sleep time (NST) (F(2,117) = 6.84, p < 0.05) and sleep efficiency (SE) (F(2,117) = 7.69, p < 0.05). Significant improvement in the sleep behaviors was observed in the experimental group using magnetic pearls. In a backward multiple regression, the effect of auricular therapy on SE after allowing for age in female participants is of high statistical significance (F(3,106) = 9.04, p < 0.001). The paper concludes that auricular therapy using magnetic pearls is an effective means of improving the quantity and quality of sleep in the elderly.

Acupuncture Points↗

Clinical observation on 25 cases of hormone dependent bronchial asthma treated by acupuncture.

The clinical observation revealed that the asthmatic symptoms in most of the patients began to be improved after several acupuncture treatments with the dosage of the drug gradually reduced. Generally, the dose of cortisone per os was decreased by 2 mg every 10 days, while that of aerosol was controlled by the patients themselves according to the condition of the disease. The symptoms in most of the patients were markedly improved after 15 treatments, but the treatment should be continued for another 10 times to consolidate the curative effect. Thus, each asthmatic patient needs to receive approximately 30 sessions of acupuncture treatment, lasting about 3 months. Thereafter, in order to prevent its relapse, the treatment should be administered 10 times each year in the summer season. It is worthy to be mentioned that, in this series, there was a female patient who suffered from anaphylactic asthma induced by dog's hair. She received 10 sessions of acupuncture treatment with no any improvement. In this case, the acupuncture treatment should not be given any longer. There were another two children patients aged 5 and 7 years respectively, for them the above method of acupuncture treatment was difficult to be used. Therefore, the auricular pressing method combined with cupping was adopted instead. The auricular points selected were Shenmen, Lung, Large Intestine, sensitive point, Pingchuan, Subcortex, and Kidney. 5 points were used each time, and pressed with the seeds of Vaccaria which were changed every 3 days. The patients were asked to press each of the points themselves 3 times a day (each time by 20 pressings). And Dazhui (Du 14), Feishu (UB 13), Pishu (UB 20) and Shenshu (UB 23) were cupped for 5 min. each time. The auricular pressing and cupping therapies may also be added for adult patients to strengthen the curative effect. The 25 cases of hormone dependent bronchial asthma were treated by acupuncture, yielding a markedly effective rate of 56% with a total effective rate of 96%, indicating that good therapeutic results can also be expected in the western countries for those cases who have already been treated with hormones.

Acupuncture Therapy↗