Cytochemical distribution of beta-glucuronidase activity in experimental brain tumors and brain tissue in vivo and in vitro

Summary Activity of beta-glucuronidase in experimental brain tumors and in nervous tissue was studied. The activity of the enzyme was detected both in gliomas and sarcomas exhibiting a high degree of anaplasia and dysplasia. Reactive glia showed high enzymatic activity; whereas activity in normal nervous tissue was absent. In tissue culture the activity of beta-glucuronidase was observed mainly in astrocytes, and its intensity expressed the degree of cellular maturation and differentiation.The investigation was carried out under a grant from PL 480 US Public Health Service Program, Agreement 05-004-1.     

Suppression of the release of prostaglandin-like substances by hydrocortisone in vivo

Muscular exercise of the dog's hind leg evokes the release of prostaglandin-like substances / PG-like substances/ into femoral venous blood. The release of PG-like substances detected by the bioassay method was significantly greater in adrenalectomized as compared to normal dogs. To test the possibility that this difference may be related to the deficiency of adrenocortical secretion in adrenalectomized dogs, the effect of hydrocortisone / HC / and aldosterone / AS / upon the release of PG-like substances induced by muscular work of the dog's hind leg was investigated. The doses of HC and AS infused intravenously or intraarterially were close to the range of physiological secretion rate of these hormones. HC suppressed the release of PG-like material by 30 to 60%, whereas AS had no effect upon the rate and duration of the release. The rate of removal of exogenous PGE₂ in the hind limb circulation was not influenced by HC, suggesting that the diminution of PG release by HC results from the suppression of PG generation rather than from the enhancement of degradation. It is suggested that inhibitory effect of HC upon the rate of the release of PG-like substances may be related to the membrane-stabilizing properties of this hormone. The difference in the intensity of the release of PG-like substances between normal and adrenalectomized dogs suggests that, at least in some conditions, the release of endogenous PGs from tissues may be influenced by the state of adrenocortical activity.     

New facts about callose events in the young ovules of some sexual and apomictic species of the Asteraceae family

Callose (β-1,3-glucan) is one of the cell wall polymers that plays an important role in many biological processes in plants, including reproductive development. In angiosperms, timely deposition and degradation of callose during sporogenesis accompanies the transition of cells from somatic to generative identity. However, knowledge on the regulation of callose biosynthesis at specific sites of the megasporocyte wall remains limited and the data on its distribution are not conclusive. Establishing the callose deposition pattern in a large number of species can contribute to full understanding of its function in reproductive development. Previous studies focused on callose events in sexual species and only a few concerned apomicts. The main goal of our research was to establish and compare the pattern of callose deposition during early sexual and diplosporous processes in the ovules of some Hieracium, Pilosella and Taraxacum (Asteraceae) species; aniline blue staining technique was used for this purpose. Our findings indicate that callose deposition accompanies both meiotic and diplosporous development of the megaspore mother cell. This suggests that it has similar regulatory functions in intercellular communication regardless of the mode of reproduction. Interestingly, callose deposition followed a different pattern in the studied sexual and diplosporous species compared to most angiosperms as it usually began at the micropylar pole of the megasporocyte. Here, it was only in sexually reproducing H. transylvanicum that callose first appeared at the chalazal pole of the megasporocyte. The present paper additionally discusses the occurrence of aposporous initial cells with callose-rich walls in the ovules of diploid species.

     

Stomatal density in Pinus sylvestris as an indicator of temperature rather than CO2: Evidence from a pan-European transect

The commonly observed negative relationship between stomatal density (SD) and atmospheric CO₂ has led to SD being proposed as an indicator of atmospheric CO₂ concentration. The use of SD as a proxy for CO₂, however, has been hampered by an insufficient understanding of the intraspecific variation of this trait. We hypothesized that SD in Pinus sylvestris, a widely distributed conifer, varies geographically and that this variation is determined by major climatic variables. By sampling needles from naturally growing trees along a latitudinal range of 32.25°, equivalent to 13.7°C gradient of mean annual temperature (MAT) across Europe, we found that SD decreased from the warmest southern sites to the coldest sites in the north at a rate of 4 stomata per mm² for each 1°C, with MAT explaining 44% of the variation. Additionally, samples from a provenance trial exhibited a positive relationship between SD and the MAT of the original localities, suggesting that high SD is an adaptation to warm temperature. Our study revealed one of the strongest intraspecific relationships between SD and climate in any woody species, supporting the utility of SD as a temperature, rather than direct CO₂, proxy. In addition, our results predict the response of SD to climate warming.     

Mechanisms involved in the anti-inflammatory action of inhaled tea tree oil in mice

Tea tree oil (TTO) is well known as an antimicrobial and immunomodulatory agent. In the present study we confirmed the anti-inflammatory properties of TTO and investigated the involvement of the hypothalamic-pituitary-adrenal (HPA) axis in the immunomodulatory action of TTO administered by inhalation. Sexually mature, 6-8-week-old, C(57)BI(10) x CBA/H (F(1)) male mice were used. One group of animals was injected intra-peritoneally (ip) with Zymosan to elicit peritoneal inflammation and was then submitted to four sessions of TTO inhalation (15 mins each). Some of the mice were simultaneously injected ip with Antalarmin, a CRH-1 receptor antagonist, to block HPA axis functions. Twenty-four hours after the injections the mice were killed by CO(2) asphyxia, and peritoneal leukocytes (PTLs) were isolated and counted. Levels of reactive oxygen species (ROS) and cyclooxygenase (COX) activity in PTLs were assessed by fluorimetric and colorimetric assays, respectively. The results obtained show that sessions of TTO inhalation exert a strong anti-inflammatory influence on the immune system stimulated by Zymosan injection, while having no influence on PTL number, ROS level, and COX activity in mice without inflammation. The HPA axis was shown to mediate the anti-inflammatory effect of TTO; Antalarmin abolished the influence of inhaled TTO on PTL number and their ROS production in mice with experimental peritonitis, but it had no effect on these parameters in mice without inflammation.     

Cholesterol conjugation potentiates the antiviral activity of an HIV immunoadhesin

Immunoadhesins are engineered proteins combining the constant domain (Fc) of an antibody with a ligand‐binding (adhesion) domain. They have significant potential as therapeutic agents, because they maintain the favourable pharmacokinetics of antibodies with an expanded repertoire of ligand‐binding domains: proteins, peptides, or small molecules. We have recently reported that the addition of a cholesterol group to two HIV antibodies can dramatically improve their antiviral potency. Cholesterol, which can be conjugated at various positions in the antibody, including the constant (Fc) domain, endows the conjugate with affinity for the membrane lipid rafts, thus increasing its concentration at the site where viral entry occurs. Here, we extend this strategy to an HIV immunoadhesin, combining a cholesterol‐conjugated Fc domain with the peptide fusion inhibitor C41. The immunoadhesin C41‐Fc‐chol displayed high affinity for Human Embryonic Kidney (HEK) 293 cells, and when tested on a panel of HIV‐1 strains, it was considerably more potent than the unconjugated C41‐Fc construct. Potentiation of antiviral activity was comparable to what was previously observed for the cholesterol‐conjugated HIV antibodies. Given the key role of cholesterol in lipid raft formation and viral fusion, we expect that the same strategy should be broadly applicable to enveloped viruses, for many of which it is already known the sequence of a peptide fusion inhibitor similar to C41. Moreover, the sequence of heptad repeat‐derived fusion inhibitors can often be predicted from genomic information alone, opening a path to immunoadhesins against emerging viruses. Copyright © 2015 European Peptide Society and John Wiley & Sons, Ltd.     

Interactions of Escherichia coli molecular chaperone HtpG with DnaA replication initiator DNA

The bacterial chaperone high-temperature protein G (HtpG), a member of the Hsp90 protein family, is involved in the protection of cells against a variety of environmental stresses. The ability of HtpG to form complexes with other bacterial proteins, especially those involved in fundamental functions, is indicative of its cellular role. An interaction between HtpG and DnaA, the main initiator of DNA replication, was studied both in vivo, using a bacterial two-hybrid system, and in vitro with a modified pull-down assay and by chemical cross-linking. In vivo, this interaction was demonstrated only when htpG was expressed from a high copy number plasmid. Both in vitro assays confirmed HtpG–DnaA interactions.