Purification and characterization of a hemagglutinin isolated from the leaves of Chenopodium (Chenopodium amaranticolor).

A hemagglutinin was isolated and purified from the leaves of Chenopodium (Chenopodium amaranticolor) using ion-exchange chromatography and affinity chromatography on fetuin-agarose matrix. It agglutinated rabbit erythrocytes. The hemagglutinin had a native molecular mass of 58 kDa, as estimated by gel filtration and showed a single band of molecular mass of 33 kDa on SDS-PAGE. It showed hemagglutination activity over the pH range 3-12 and was found to be stable up to 70 degrees C. On isoelectrofocussing, the pI of this hemagglutinin was estimated to be 5.25. However, it was found to contain seven charge variants when isoelectrofocussing was performed in presence of 6M urea.     

Localization and biological activities of melatonin in intact and diseased gastrointestinal tract (GIT).

Melatonin (MT), an indole formed enzymatically from L-trytophan (Trp), was first discovered in the bovine pineal gland in 1958 by Lerner et al. Melatonin is the most versatile and ubiquitous hormonal molecule produced not only in the pineal gland but also in various other tissues of invertebrates and vertebrates, particularly in the gastrointestinal tract (GIT). This review focuses on the localization, production, metabolism and the functions of MT in GIT and the duodenal unit (liver, biliary routes and pancreas), where multi-step biosynthetic pathways of this indole, similar to those in pinealocytes, have been identified. These biosynthetic steps of MT, including two major rate limiting enzymes; arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O-methyltransferase (HIOMT), transforming L-tryptophan (Trp), originally identified in pinealocytes, have been also detected in entero-endocrine (EE) cells of GIT, where this indole appears to act in endocrine, paracrine and/or luminal pathway directly or through G-protein coupled MT receptors. Studies of the distribution of MT in GIT mucosa showed that this indole is generated in GIT in much larger amounts than it is produced in the pineal gland. Melatonin acts in GIT, partly locally in paracrine fashion and is partly released into portal circulation, to be taken up by the liver. It is then metabolized and excreted with the bile to small bowel and finally returns to liver through entero-hepatic circulation. The production of MT by the pineal gland shows circadian rhythm with high night-time surge, especially at younger age, followed by the fall during the day-light time. As a highly lipophylic substance, MT reaches all body cells within minutes, thus, serving as a convenient circadian timing signal. Following pinealectomy, the light/dark cycle of plasma MT levels disappears, while its day-time blood concentration is maintained mainly due to its release from the GIT. According to our experience, after oral application of Trp, the plasma MT increases in dose-dependent manner both in intact and pinealectomized animals and humans, indicating that GIT but not the pineal gland is a source of this indole. In GIT MT exhibits a wide spectrum of activities such as circadian entrainment, antioxidant and free radicals scavenging activity, Melatonin (MT), an indole formed enzymatically from L-trytophan (Trp), was first discovered in the bovine pineal gland in 1958 by Lerner et al. Melatonin is the most versatile and ubiquitous hormonal molecule produced not only in the pineal gland but also in various other tissues of invertebrates and vertebrates, particularly in the gastrointestinal tract (GIT). This review focuses on the localization, production, metabolism and the functions of MT in GIT and the duodenal unit (liver, biliary routes and pancreas), where multi-step biosynthetic pathways of this indole, similar to those in pinealocytes, have been identified. These biosynthetic steps of MT, including two major rate limiting enzymes; arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O-methyltransferase (HIOMT), transforming L-tryptophan (Trp), originally identified in pinealocytes, have been also detected in entero-endocrine (EE) cells of GIT, where this indole appears to act in endocrine, paracrine and/or luminal pathway directly or through G-protein coupled MT receptors. Studies of the distribution of MT in GIT mucosa showed that this indole is generated in GIT in much larger amounts than it is produced in the pineal gland. Melatonin acts in GIT, partly locally in paracrine fashion and is partly released into portal circulation, to be taken up by the liver. It is then metabolized and excreted with the bile to small bowel and finally returns to liver through entero-hepatic circulation. The production of MT by the pineal gland shows circadian rhythm with high night-time surge, especially at younger age, followed by the fall during the day-light time. As a highly lipophylic substance, MT reaches all body cells within minutes, thus, serving as a convenient circadian timing signal. Following pinealectomy, the light/dark cycle of plasma MT levels disappears, while its day-time blood concentration is maintained mainly due to its release from the GIT. According to our experience, after oral application of Trp, the plasma MT increases in dose-dependent manner both in intact and pinealectomized animals and humans, indicating that GIT but not the pineal gland is a source of this indole. In GIT MT exhibits a wide spectrum of activities such as circadian entrainment, antioxidant and free radicals scavenging activity, cytoprotective, anti-inflammatory and healing efficacy of various GIT lesions such as esophagitis, gastritis, peptic ulcer, pancreatitis and colitis. This review concentrates on the generation and pathophysiological implication of MT in GIT and related organs.     

Characterization of biological activities of Brucella melitensis lipopolysaccharide

Biological activities of lipopolysaccharide (LPS) from Brucella melitensis 16M were characterized in comparison with LPS from Escherichia coli O55. LPS extracted from B. melitensis was smooth type by electrophoretic analysis with silver staining. The endotoxin-specific Limulus activity of B. melitensis LPS was lower than that of E. coli LPS. There was no significant production of tumor necrosis factor-alpha and nitric oxide in RAW 264.7 macrophage cells stimulated with B. melitensis LPS, although E. coli LPS definitely induced their production. On the other hand, B. melitensis LPS exhibited a higher anti-complement activity than E. coli LPS. B. melitensis LPS as well as E. coli LPS exhibited a strong adjuvant action on antibody response to bovine serum. The characteristic biological activities of B. melitensis are discussed.     

Anti-substance P anti-idiotypic antibodies. Characterization and biological activities

Antibodies to substance P (SP) produced in rabbits have been characterized for their specificity toward SP and some 30 SP-related peptides. For each compound, we observed a close correlation between capacity of binding to anti-SP antibodies and biological activity, namely their spasmogenic effect on guinea pig ileum in vitro and their hypotensive effect in the rat in vivo, indicating that the combining sites of anti-SP and SP receptor(s) are structurally very similar. Further immunization of five rabbits with anti-SP immunoglobulins elicited in two allotype-matched animals the production of anti-SP anti-idiotypic antibodies. These latter antibodies were found to strongly inhibit the spasmogenic action of SP on the guinea pig ileum. In contrast, they specifically enhanced, like SP, phospholipid turnover in rat parotid gland cells, a physiological function mediated through an activation of SP receptors. Immunocytochemical studies actually revealed the presence of specific membranous binding sites for anti-idiotypic antibodies on the parotid gland-dissociated cells. The anti-idiotypic antibodies described here, which thus behave either as agonists or antagonists for SP depending on the biological test, might be used as original and powerful tools not only in studies of the receptor stereospecificity but also in attempts to purify the membranous SP receptors.     

Anatomical and hnctional differences and nyctinastic leaf movements in Chenopodium album L. and Chenopodium hircinum Schrad. (Chenopodiaceae)

The anatomical structure of the leaves of Chenopodium album L. and Chenopodium hircinum Schrad. has been examined. Leaf chlorophyll content (Chl), specific leaf area (SLA), specific leaf weight (SLA^-1) and nitrogen content (N₂) were estimated in both species. Ch1, SLA and N₂ were greater in C. album than in C. hircinum. SLA^-1 data showed that C. album is more efficient than C. hircinum , because the former invests smaller quantities of dry weight to achieve a square centimetre of leaf area. The possible effects of Chl, N₂ and leaf anatomy on efficient gas exchange are discussed. The presence of nyctinastic leaf nocturnal movements was detected in both species. The meaning of this is discussed in relation to the prevention of the loss of pollen grains.     

Removal of carbohydrate from influenza A virus and its hemagglutinin and the effect on biological activities.

Treatment of influenza virus and its purified hemagglutining with glycosidases from Diplococcus pneumoniae, which included beta-galactosidase, beta-N-acetylglucosminidase, and endoglycosidase D, released amino and neutral sugars from the virus and these as well as large oligosaccharides from the purified hemagglutinin. The released glucosamine-containing oligosaccharides were of one discrete size. Large oligosaccharides not removed by the glycosidases were found on the virus as well as the hemagglutinin. Some oligosaccharides on the virus were inaccessible to the enzymes, since they could be removed only from the purified hemagglutinin. Approximately 50% of the hemagglutinin carbohydrates could be removed without effect on hemagglutinating activity. Similarly, removal of 20 to 25% of the carbohydrates from intact virus particles did not alter infectivity.     

Characterization of murine interferon-alpha 12 (MuIFN-alpha12): biological activities and gene expression

Interferon alpha (IFN-alpha) belongs to the type I interferon family and consists of multiple subtypes in many species. In the mouse, there are at least 14 IFN-alpha genes and 3 IFN-alpha pseudogenes, the most recently identified of which are murine interferon-alpha 12 (MuIFN-alpha12), MuIFN-alpha13 and MuIFN-alpha14. To further study the biological activities of MuIFN-alpha12, we have produced a recombinant MuIFN-alpha12 (rMuIFN-alpha12) protein using COS-1 cells. rMuIFN-alpha12 was found to inhibit the growth of murine myeloid leukemia JCS cells. Flow cytofluorometric analysis with propidium iodide staining showed that the growth inhibitory activity of rMuIFN-alpha12 may be caused by the induction of apoptosis. Flow cytofluorometric analysis also revealed that rMuIFN-alpha12 was able to up-regulate the expression of MHC-I on both JCS cells and primary macrophages. Functional studies indicated that a MuIFN-alpha12 transgene could induce an anti-viral state in L929 cells against Influenza A virus. Moreover, expression of MuIFN-alpha12 was not detectable by RT-PCR in untreated, Influenza A virus infected, polyI:polyC induced L929 cells, or in a wide range of normal murine tissues. Taken together, this data shows that MuIFN-alpha12 is a protein with all the biological traits of a type I IFN.     

土壤节肢动物对箭竹凋落叶分解过程中酶活性的影响

为了解凋落物分解过程中土壤节肢动物与土壤酶活性的相互联系,以川西亚高山森林箭竹(Fargesia spathacea)凋落叶为对象,通过原位控制实验,于2016年4月至2018年4月研究了土壤节肢动物对凋落叶分解过程中碳、氮和磷转化相关酶活性的影响。结果表明:生物抑制剂施用降低了分解袋中土壤节肢动物49.7%～66.8%的个体密度和19.2%～46.3%的类群数量;对照和处理分解袋中凋落叶碳、氮和磷转化相关酶活性随分解过程呈现相似的动态;与处理相比,土壤节肢动物参与(对照)显著提高了凋落叶分解过程中蔗糖酶、β-葡聚糖苷酶、纤维素酶、多酚氧化酶、过氧化物酶、N-乙酰-β-D-氨基葡萄糖苷酶和酸性磷酸酶活性;土壤节肢动物对凋落叶分解过程中酶活性的贡献率在达到一个明显的峰值后快速降低;土壤温度和土壤节肢动物类群数量与蔗糖酶活性呈显著正相关,与β-葡聚糖苷酶、纤维素酶、多酚氧化酶、过氧化物酶、N-乙酰-β-D-氨基葡萄糖苷酶和酸性磷酸酶活性呈显著负相关。土壤节肢动物对凋落叶分解过程中酶活性促进效应随酶类型和分解时间变化存在差异,与土壤节肢动物群落结构和分解环境密切相关。     

Biological activities and chemistry of saponins from Chenopodium quinoa Willd.

Chenopodium quinoa Willd. is a valuable food source which has gained importance in many countries of the world. The plant contains various bitter-tasting saponins which present an important antinutritional factor. Various triterpene saponins have been reported in C. quinoa including both monodesmosidic and bidesmosidic triterpene saponins of oleanolic acid, hederagenin, phytolaccagenic acid, and serjanic acid as the major aglycones and other aglycones as 3β-hydroxy-23-oxo-olean-12-en-28-oic acid, 3β-hydroxy-27-oxo-olean-12-en-28-oic acid, and 3β, 23α, 30β-trihydroxy-olean-12-en-28-oic acid. A tridesmosidic saponin of hederagenin has also been reported. Here we review the occurrence, analysis, chemical structures, and biological activity of triterpene saponins of C. quinoa. In particular, the mode of action of the mono- and bidesmosidic triterpene saponins and aglycones are discussed.     

Biological activities of extracts from Chenopodium ambrosioides Lineu and Kielmeyera neglecta Saddi

ABSTRACT: BACKGROUND: Chenopodium ambrosioides and Kielmeyera neglecta are plants traditionally used in Brazil to treat various infectious diseases. The study of the biological activities of these plants is of great importance for the detection of biologically active compounds. METHODS: Extracts from these plants were extracted with hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc) and ethanol (EtOH) and assessed for their antimicrobial properties, bioactivity against Artemia salina Leach and antifungal action on the cell wall of Neurospora crassa. RESULTS: Extracts from C. ambrosioides (Hex, DCM and EtOH) and K. neglecta (EtOAc and EtOH) showed high bioactivity against A. salina (LD50 < 1000 ug/mL), which might be associated with cytotoxic activity against cancer cells. C. ambrosioides Hex and DCM showed specific activity against yeasts, highlighting the activity of hexanic extract against Candida krusei (MIC = 100 ug/mL). By comparing the inhibitory concentration of 50% growth (IC 50%) with the growth control, extracts from K. neglecta EtOAc and EtOH have shown activities against multidrug-resistant bacteria (Enterococcus faecalis ATCC 51299 and Staphylococcus aureus ATCC 43300), with IC 50% of 12.5 ug/mL The assay carried out on N. crassa allowed defining that extracts with antifungal activity do not have action through inhibition of cell wall synthesis. CONCLUSIONS: Generally speaking, extracts from C. ambrosioides and K. neglecta showed biological activities that have made the search for bioactive substances in these plants more attractive, illustrating the success of their use in the Brazilian folk medicine.     

Effect of the addition of oligosaccharides on the biological activities and antigenicity of influenza A/H3N2 virus hemagglutinin

Influenza A/H3N2 viruses have developed an increased number of glycosylation sites on the globular head of the hemagglutinin (HA) protein since their appearance in 1968. Here, the effect of addition of oligosaccharide chains to the HA of A/H3N2 viruses on its biological activities was investigated. We constructed seven mutant HAs of A/Aichi/2/68 virus with one to six glycosylation sites on the globular head, as found in natural isolates, by site-directed mutagenesis and analyzed their intracellular transport, receptor binding, and cell fusion activities. The glycosylation sites of mutant HAs correspond to representative A/H3N2 isolates (A/Victoria/3/75, A/Memphis/6/86, or A/Sydney/5/97). The results showed that all the mutant HAs were transported to the cell surface as efficiently as wild-type HA. Although mutant HAs containing three to six glycosylation sites decreased receptor binding activity, their cell fusion activity was not affected. The reactivity of mutant HAs having four to six glycosylation sites with human sera collected in 1976 was much lower than that of wild-type HA. Thus, the addition of new oligosaccharides to the globular head of the HA of A/H3N2 viruses may have provided the virus with an ability to evade antibody pressures by changing antigenicity without an unacceptable defect in biological activity.     

Synthesis and biological activities of 8(14)a-homocalcitriol.

8(14)a-Homocalcitriol was synthesized and tested for its biologic activities. It exhibited a vitamin D agonist activity profile. The compound was bound to the pig intestinal receptor with an affinity slightly less than calcitriol, showed the same potency in inducing HL 60 cell differentiation and inhibition of keratinocyte proliferation as calcitriol, and was found to be approximately 10-fold less potent in inducing hypercalcemia and hypercalciuria after a single injection in normal rats.     

Cross-protection between arabis mosaic virus and grapevine fan leaf virus isolates in Chenopodium quinoa

Cross-protection experiments were performed in Chenopodium quinoa using arabis mosaic virus (ArMV) and grapevine fanleaf virus (GFLV) isolates. Two factors were specially studied, namely the time interval and the distance between the two inoculations, respectively, with the hypovirulent isolate and with the hyper virulent challenge isolale. ArMV-S clearly protected C. quinoa from a super infection with GFLV-F13 as shown by a diminution, or even suppression, of the synthesis of the coat protein and the nucleic acids of the GFLV-F13 isolate. In the homologous interaction between GFLV isolates (GH and F13), protection was also observed. In the interaction between GFLV-GH and ArMV-862, by contrast, symptoms were typical of the hyper virulent ArMV-862 and the amount of coat protein of ArMV-862 was normal.     

Analysis and biological activities of anthocyanins

Anthocyanins are naturally occurring compounds that impart color to fruits, vegetables, and plants. They are probably the most important group of visible plant pigments besides chlorophyll. Apart from imparting color to plants, anthocyanins also have an array of health-promoting benefits, as they can protect against a variety of oxidants through a various number of mechanisms. However, anthocyanins have received less attention than other flavonoids, despite this. This article reviews their biological functions and pre-clinical studies, as well as the most recent analytical techniques concerning anthocyanin isolation and identification.     

Characterization of influenza hemagglutinin interactions with receptor by NMR

In influenza, the envelope protein hemagglutinin (HA) plays a critical role in viral entry by first binding to sialic acid receptors on the cell surface and subsequently mediating fusion of the viral and target membranes. In this work, the receptor binding properties of influenza A HA from different subtypes (H1 A/California/04/09, H5 A/Vietnam/1205/04, H5 A/bar-headed goose/Qinghai/1A/05, and H9 A/Hong Kong/1073/99) have been characterized by NMR spectroscopy. Using saturation transfer difference (STD) NMR, we find that all HAs bind to the receptor analogs 2,3-sialyllactose and 2,6-sialyllactose, with subtle differences in the binding mode. Using competition STD NMR, we determine the receptor preferences for the HA subtypes. We find that H5-Qinghai and H9-Hong Kong HA bind to both receptor analogs with similar affinity. On the other hand, H1 exhibits a clear preference for 2,6-sialyllactose while H5-Vietnam exhibits a clear preference for 2,3-sialyllactose. Together, these results are interpreted within the context of differences in both the amino acid sequence and structures of HA from the different subtypes in determining receptor preference.