α1-antitrypsin and its C-terminal fragment attenuate effects of degranulated neutrophil-conditioned medium on lung cancer HCC cells, in vitro

Background  

Tumor microenvironment, which is largely affected by inflammatory cells, is a crucial participant in the neoplastic process through promotion of cell proliferation, survival and migration. We measured the effects of polymorphonuclear neutrophil (PMN) conditioned medium alone, and supplemented with serine proteinase inhibitor α-1 antitrypsin (AAT) or its C-terminal fragment (C-36 peptide), on cultured lung cancer cells.     

Obtaining and characterization of “Holospora curviuscula” and Holospora obtusa, bacterial symbionts of the macronuclei of Paramecium bursaria and Paramecium caudatum

The symbionts of the macronuclei of Paramecium bursaria and P. caudatum, “Holospora curviuscula” 02AZ16 and H. obtusa 88Ti, respectively, were obtained and investigated. The 16S rDNA nucleotide sequences of “Holospora curviuscula” were obtained for the first time. The differences in 16S rDNA (3.4%) suggest their classification within the genus Holospora. Molecular phylogenetic analysis of the symbionts revealed that these intranuclear symbionts of the ciliates belonged to the order Rickettsiales, forming within a compact cluster of related species.     

Human mast cells decrease SLPI levels in type II – like alveolar cell model, in vitro

Background  

Mast cells are known to accumulate at sites of inflammation and upon activation to release their granule content, e.g. histamine, cytokines and proteases. The secretory leukocyte protease inhibitor (SLPI) is produced in the respiratory mucous and plays a role in regulating the activity of the proteases.     

Effects of β-lactam antibiotics, auxins, and cytokinins on shoot regeneration from callus cultures of two hybrid aspens, Populustremuloides?×?P. tremula and P. xcanescens?×?P. gradidentata

The effects of β-lactam antibiotics (penicillin, carbenicillin and cefotaxime), cytokinins, and auxins including phenylacetic acid, a β-lactam breakdown product, were evaluated during in vitro shoot morphogenesis in two hybrid aspens; P. tremuloides × P. tremula (XTTa) and P. x canescens × P. grandidentata (XCaG). Although different callus and shoot induction media were used for both hybrids, the β-lactams often engendered similar responses. At concentrations of 1,000 mg l⁻¹, carbenicillin adversely impacted shoot elongation and, to a lesser degree, shoot regeneration. Cefotaxime enhanced caulogenesis for all of the concentrations evaluated (125–500 mg l⁻¹) especially when the cytokinin thidiazuron was used for shoot induction. The shoots formed faster and in greater numbers; and the improvements were significant (α = 0.05) for both hybrids. However, hyperhydricity was more problematic when cefotaxime was included in the media. The incidence of shoot hyperhydricity for the XCaG hybrid was more than twice as great for the highest cefotaxime concentration evaluated (500 mg l⁻¹) than for the control (>90% vs. ~40%). Penicillin had an opposite effect. Hyperhydricity frequencies for the XCaG hybrid were lower when the media were supplemented with penicllin and the reductions were statistically significant at concentrations of 500–1,000 mg l⁻¹. The effects of the antibiotics were generally not reproduced by the auxins (0.1–100 μM), including phenylacetic acid, or the other potential β-lactam degradation products evaluated (e.g. phenylmalonic acid, aminopenicillanic acid). The antibiotics may have affected shoot hyperhydicity indirectly via changes in the time course of shoot regeneration.     

The mutants compacta ?hnlich, Nitida and Grandiflora define developmental compartments and a compensation mechanism in floral development in Antirrhinum majus

In order to improve our understanding of floral size control we characterised three mutants of Antirrhinum majus with different macroscopic floral phenotypes. The recessive mutant compacta ?hnlich has smaller flowers affected mainly in petal lobe expansion, the dominant mutant Grandiflora has overall larger organs, whilst the semidominant mutation Nitida exhibits smaller flowers in a dose-dependent manner. We developed a cell map in order to establish the cellular phenotypes of the mutants. Changes in organ size were both organ- and region-specific. Nitida and compacta ?hnlich affected cell expansion in proximal and distal petal regions, respectively, suggesting differential regulation between petal lobe regions. Although petal size was smaller in compacta ?hnlich than in wild type, conical cells were significantly bigger, suggesting a compensation mechanism involved in petal development. Grandiflora had larger cells in petals and increased cell division in stamens and styles, suggesting a relationship between genes controlling organ size and organ identity. The level of ploidy in petals of Grandiflora and coan was found to be equivalent to wild type petals and leaves, ruling out an excess of growth via endoreduplication. We discuss our results in terms of current models about control of lateral organ size.     

Regulation of IFN-γ and IL-10 synthesis in vivo, as well as continuous antigen exposure, is associated with tolerance to murine skin allografts

C3H/HEJ mice are rendered hyporesponsive to multiple minor incompatible (B10.BR) skin allografts by pretreatment with irradiated B10.BR lymphoid cells injected via the portal vein, but not the lateral tail vein. As assessed by PCR with lymphocytes taken from grafted mice, or by measuring cytokines in vitro from antigen-restimulated cells, this hyporesponsiveness is associated with decreased mRNA for IFN-γ and IL-2 production, but enhanced mRNA for IL-4 and IL-10 production. In mice given B10.BR cells via the tail vein, but in addition injected every second day with anti-IFN-γ antibody, similar enhanced graft survival (with diminished IFN-γ/IL-2 and enhanced IL-4/IL-10 production) was seen. In a separate study spleen cells from pretreated mice were “parked” in lethally irradiated syngeneic mice for 21 days, along with B10.BR skin grafts to some of the recipients. Only when recipients received this reexposure to B10.BR antigen did adoptively transferred spleen cells show “persistence” of the ability to produce delayed graft rejection and preferential IL-4 production in vitro.     

Tumour necrosis factor activates the mitogen-activated protein kinases p38α and ERK in the synovial membrane in vivo

Tumour necrosis factor (TNF) is considered to be a major factor in chronic synovial inflammation and is an inducer of mitogen-activated protein kinase (MAPK) signalling. In the present study we investigated the ability of TNF to activate MAPKs in the synovial membrane in vivo. We studied human TNF transgenic mice – an in vivo model of TNF-induced arthritis – to examine phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun amino terminal kinase (JNK) and p38MAPKα in the inflamed joints by means of immunoblot and immunohistochemistry. In addition, the effects of systemic blockade of TNF, IL-1 and receptor activator of nuclear factor-κB (RANK) ligand on the activation of MAPKs were assessed. In vivo, overexpression of TNF induced activation of p38MAPKα and ERK in the synovial membrane, whereas activation of JNK was less pronounced and rarely observed on immunohistochemical analysis. Activated p38MAPKα was predominantly found in synovial macrophages, whereas ERK activation was present in both synovial macrophages and fibroblasts. T and B lymphocytes did not exhibit major activation of any of the three MAPKs. Systemic blockade of TNF reduced activation of p38MAPKα and ERK, whereas inhibition of IL-1 only affected p38MAPKα and blockade of RANK ligand did not result in any decrease in MAPK activation in the synovial membrane. These data indicate that TNF preferentially activates p38MAPKα and ERK in synovial membrane exposed to TNF. This not only suggests that targeted inhibition of p38MAPKα and ERK is a feasible strategy for blocking TNF-mediated effects on joints, but it also shows that even currently available methods to block TNF effectively reduce activation of these two MAPKs.     

Eulophidae (hymenoptera) parasitoids of Phyllonorycter apparella and phyllonorycter populifoliella (Lepidoptera, Gracillariidae), pests of Populus tremula and populus nigra in Ul’yanovsk province, Russia

Parasitoid complexes of two species of the genus Phyllonorycter (Ph. apparella and Ph. populifoliella) reared from 2 plants (Populus tremula and P. nigra) were studied in Ul’yanovsk Province, Russia. Twenty species of parasitoids are new for Ph. apparella and 12 are new for Ph. populifoliella. Minotetrastichus frontalis and Pnigalio agraules are the dominant species for both parasitoid complexes. The highest relative abundance was observed for Ph. apparella on Populus tremula (75%) and for Ph. populifoliella on P. nigra (61%). The parasitoid complexes of Ph. apparella and Ph. populifoliella show high similarity (the Jaccard coefficient is 0.65).     

The in vivo effects of neutralizing antibodies against IFN-γ, IL-4, or IL-10 on the humoral immune response in young and aged mice

In the present study we investigated whether age-related changes in the composition and functional properties of murine CD4⁺ T cells are reflected in vivo by a changed humoral response to influenza vaccine in aged mice. After the primary immunization, the titers of influenza-specific IgM, IgG1, IgG2a, and IgG2b, but not of IgG3 and IgE, were significantly reduced in aged mice compared to young mice. Treatment of aged mice with anti-IFN-γ, anti-IL-4, or anti-IL-10 resulted in levels of IgM and IgG1 comparable to those found in young mice, whereas IgG2a and IgG2b were further decreased. After the booster immunization IgE was significantly enhanced in aged mice, whereas no differences were observed with regard to the other isotypes. During the primary response in young mice, anti-IFN-γ stimulated IgG1 and IgE, whereas an inhibition of IgG2a, IgG2b, and IgG3 was observed. Anti-IL-4 caused a decrease only in IgG3 while anti-IL-10 increased IgM and IgG1 and decreased IgG2b and IgG3. During the primary response in aged mice, all anti-cytokine antibodies enhanced IgM and IgG1 while IgE was only enhanced by anti-IL-10. By contrast, IgG3 was inhibited by anti-IFN-γ and anti-IL-10. Anti-cytokine treatment of young mice increased all isotypes, except IgG3, in the secondary response, whereas the secondary response in aged mice was largely insensitive to anti-cytokine treatment. These data therefore support the idea that the in vivo effects of cytokines on isotype switching are dependent on the differentiation stage of B cells which may be different in young and aged mice.     

Comparative study (AFLP and morphology) of three species of Prosopis of the Section Algarobia: P. juliflora, P. pallida, and P. limensis. Evidence for resolution of the “P. pallida–P. juliflora complex”

The problems of delimitation of species of Prosopis originate from the few morphological discontinuities which exist among some of them; some, however, originated as a result of wide distribution of germplasm without proper knowledge of the species, in particular, much material catalogued as P. juliflora, but being of other species, was distributed for reforestation projects worldwide. This work tests the morphological results obtained for P. pallida and P. limensis of the Peruvian–Ecuadorian coast and for P. juliflora of the Caribbean Basin of Colombia and Venezuela utilizing a study of AFLPs and a study of the morphology of plantlets developed in a conventional garden study. The phenogram obtained for the AFLPs demonstrates each of the three species to be a well differentiated cluster and the molecular variance between them is significantly greater than the variance within each species. Study of the plantlets also indicates statistically significant differences for four morphological characters between P. juliflora and the other two species (P. pallida and P. limensis). These results, in addition to the morphological differentiation evident between adult plants of P. pallida and P. limensis and the clear separation of these two species from P. juliflora, corroborate the genetic identity of the three taxa analyzed.     

Synthesis and transporter binding properties of (R)-2β,3β- and (R)-2α,3α-diaryltropanes

(R)-2-Aryl-2-tropinone (9) was synthesized from (R)-2-carbomethoxy-3-tropinone (5) and was used as the key intermediate for the synthesis of (R)-2β,3β- and (R)-2α,3α-diaryltropanes. Inhibition of radioligand binding studies at the dopamine, serotonin, and norepinephrine transporters showed that the (R)-3β-(4-methylphenyl)-2β-phenyltropane (3b, RTI-422) possessed an IC₅₀ value of 1.96 nM at the dopamine transporter and was highly selective for this transporter relative to the serotonin and norepinephrine transporters.     

In?vitro evaluation of the effect of the nematophagous fungi Duddingtonia flagrans , Monacrosporium sinense and Pochonia chlamydosporia on Schistosoma mansoni eggs

The in vitro effect of four isolates of the nematophagous fungi Duddingtonia flagrans (AC 001), Monacrosporium sinense (SF 53) and Pochonia chlamydosporia (VC 1 and VC 4) on eggs of Schistosoma mansoni was examined. One thousand S. mansoni eggs were plated on 2% water–agar with the grown isolates and control without fungus. After 7, 14 and 21 days, the eggs were removed and classified according to the following parameters: type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, besides hyphal penetration and internal egg colonization. Significant differences (P < 0.01) were found among the studied fungal isolates for ovicidal activity, confirming type 3 effect for the isolates VC 1 and VC 4, which characterizes the ovicidal activity of a fungus. Type 3 effect was only found for P. chlamydosporia (VC 1 and VC 4) with 26.6 and 17.2%, 25.6 and 22.6%, 27.4 and 23.9% in the 7, 14 and 21 days respectively (P < 0.01). P. chlamydosporia can thus be a potential biological control agent for S. mansoni eggs.     

Interaction between cadmium, zinc and silver-substituted plastocyanin and cytochrome b 6 f complex — heavy metals toxicity towards photosynthetic apparatus

This paper reports the results of research on the interaction between the cytochrome f of the active cytochrome b ₆ f complex (incubated with Cd-, Zn-, and Ag-substituted plastocyanins) and Cu-plastocyanin. The presented studies show, that the metal derivatives of plastocyanin can have an influence on the photosynthetic electron transfer path: cytochrome b ₆ f complex — photosystem I. The metal-substituted plastocyanins occupy the plastocyanin electron transfer site of the cytochrome f. The stopped-flow measurements show, that although the metal derivatives of plastocyanin do not influence the rate of cyt f- Pc electron transfer, creation of the non-electron-transfer complexes characterised by a strong binding between the cyt f and substituted plastocyanins and their slow release, dependent on the redox state of the substituted metal, results in the decrease of a turnover of the cytochrome complex. The research was done in the Department of Plant Biochemistry, Freiburg University, Sch?nzlestrasse 1, 79 104 Freiburg, Germany     

Epigenetic reprogramming,gene expression and in vitro development of porcine SCNT embryos are significantly improved by a histone deacetylase inhibitor—m-carboxycinnamic acid bishydroxamide (CBHA)

Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer (SCNT). Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treatment with various histone deacetylase inhibitors (HDACi), including Trichostatin A, Scriptaid, and m-carboxycinnamic acid bishydroxamide (CBHA), in which only the effect of CBHA has not yet been tested in other species. In this paperweexamine the effect ofCBHAtreatment on the development of porcine SCNT embryos. We have discovered the optimum dosage and time for CBHA treatment: incubating SCNT embryos with 2 μmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 (AcH3K9), acetylation at histone 3 lysine 18 (AcH3K18), and acetylation at histone 4 lysine 16 (AcH4K16) was raised after CBHAtreatment. Meanwhile,CBHAtreatment improved the expression of development relating genes such as pou5f1, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was not significantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNTembryos can be modified by CBHA treatment.     

Comprehensive comparisons of three pennate diatoms, Diatoma tenuae, Fragilaria vaucheriae, and Navicula pelliculosa, isolated from summer Arctic reservoirs (Svalbard 79°N), by fine-scale morphology and nuclear 18S ribosomal DNA

Here we report morphological and molecular characteristics of dominant freshwater diatoms in summer Arctic reservoirs of Svalbard (Norway), using four culture isolates, when we collected the samples in the field on 15 August 2005. Analyses of morphology and BLAST searches with 18S rDNA sequences identified them to Diatoma tenue (HYNP006, HYNP013), Navicula pelliculosa (HYNP021), and Fragilaria vaucheriae (HYNP022), respectively. Comparative studies of morphology revealed that the body shapes of the three polar diatoms were nearly identical to the known morphology of each species; however, they were considerably shorter in body length than previously described identical species from other locations. The 18S rDNA sequences of the diatoms were nearly identical to the same species from temperate and other regions. Phylogenetic analysis showed that the polar diatoms each formed a clade with their identical species and genera according to their taxonomic positions. This suggests that the polar diatoms may possess little or no genetic or morphological variation compared to more temperate strains.     

A comparative description and distribution of the flying fishes—Cypselurus poecilopterus, C. simus, and C. callopterus, sorted out into the species group of spotwing species of the subgenus Poecilocypselurus

The three species of flying fishes—Indo-Pacific Cypselurus poecilopterus, Central Pacific C. simus, and Eastern Pacific C. callopterus form a particular species group within the subgenus Poecilocypselurus. These species rather weakly differ in the body height, number of predorsal scales and number of gill rakers. The sculls of these three species are very much alike; there are just very small differences in the proportions of some bones and the number of skull lateral line canals and pores. It could be imagined that C. poecilopterus has appeared in the Indo-Malayan Archipelago area and thence extended in all directions (up to the coasts of the Eastern Africa, Northern and Eastern Australia, New Guinea and adjacent islands, the western islands of Polynesia and the waters of Japan). C. callopterus inhabiting warm waters of the Eastern Pacific and C. simus whose area is situated in the central part of the Pacific Ocean are seemingly derived from this species.     

Takeoff flight performance and plumage wettability in Cassin’s Auklet Ptychoramphus aleuticus, Xantus’s Murrelet Synthliboramphus hypoleucus and Leach’s Storm-petrel Oceanodroma leucorhoa

Due to their marine habitats and distinctive foraging modes, seabirds face unique challenges with respect to flying that are negotiated differently by various species. One such challenge is taking off from the water with wet plumage. This study evaluated plumage wettability and takeoff performance in three seabird species: two wing-propelled divers with high wing loading, Cassin’s Auklet Ptychoramphus aleuticus and Xantus’s Murrelet Synthliboramphus hypoleucus; and Leach’s Storm-petrel Oceanodroma leucorhoa, a surface feeder with low wing loading. The plumages of the diving birds held less water than that of O. leucorhoa (~6.7% of body mass vs 9.5%). This difference is explained by O. leucorhoa’s surface to volume ratio being larger than that of the alcids. Furthermore, the alcids have afterfeathers larger than those of O. leucorhoa, which promotes a better insulation during diving. Examination of takeoff performance both before and after experimentally submerging the birds indicated that wingbeat frequency, speed and mass-specific power (peak and mean), and energy per wingbeat decreased in all species when plumage was experimentally wetted, whereas mean acceleration increased. O. leucorhoa was more strongly affected by wet plumage than the alcids, with a 32% of reduction in mass-specific energy per wingbeat compared to ≤25% in the alcids. Takeoff angle was reduced in alcids, but not significantly so in O. leucorhoa. Our results offer insights into the takeoff mechanics problems of wet seabirds given their differences in life history and morphology.     

Highly differentiated population structure of a Mangrove species, Bruguiera gymnorhiza (Rhizophoraceae) revealed by one nuclear GapCp and one chloroplast intergenic spacer trnF–trnL

To evaluate the genetic diversity of a mangrove species and clarify the genetic structure of its populations, we studied nucleotide polymorphism in two DNA regions of Bruguiera gymnorhiza collected from the southern islands of Japan, Thailand, Malaysia, Indonesia, Micronesia, and India. The two DNA sequences were the chloroplast (cp) intergenic spacer between trnL and trnF genes (ca. 300 bp), and a part (ca. 550 bp) of the nuclear gene coding for glyceraldehyde-3-phosphate dehydrogenase (GapCp). Little polymorphism was found within each of the three geographical regions, Pacific Ocean, Bay of Bengal and Arabian Sea. Throughout the vast regions east of the Malay peninsula including Indonesia, Thailand, Micronesia and the southern islands of Japan (Pacific Ocean), essentially only one haplotype (apart from variation in number of a T repeat) was present. A second haplotype was present on the western coast of Malay Peninsula and the eastern coast of India (Bay of Bengal). On the southwest of Malay Peninsula both of these haplotypes were present. Finally a third haplotype was found only on the western coast of India (Arabian Sea). When taken over all geographic populations, total nucleotide variation within the species was large (μ = 0.006, average of the two genes). Our results are consistent with the hypothesis that this low genetic diversity within any local population and differentiation between the different oceans or regions are caused by very low gene flow between each of the different oceans coupled with frequent fluctuation of population sizes due to the change in sea level. The significance of these results is discussed from evolutionary point of the mangrove forests.