Purification of recombinant proteins with an example of tumor necrosis factor thymosin-α1

Hybrid protein, cancer necrosis factor thymosin-α1 (TNF-T), when synthesizing in strain-producer of Escherichia coli SG200-50 with plasmid pThy315, was a part of “inclusion bodies” mostly in the form of a high-molecular complex with other proteins due to the S-S bonds formation. An approach of purification of TNF-T has been proposed, which is based on the destruction of the complex in the presence of sodium dodecylsulfate (DDS-Na) and dithiotreitol (DDT) followed by gel-filtration on Sephadex G-100 and renaturation by ultrafiltration on hollow fibers. The method allows the isolation of electrophoretically homogeneous TNF-T containing no DDS-Na and having high cytotoxic activity against cancer cells of mouse adenocarcinome L-929. The yield of TNF-T achieved 80% relative its content in biomass and 30% relative the total protein.     

Different abilities of Friend murine leukemia virus (MuLV) and Moloney MuLV to induce promonocytic leukemia are due to determinants in both psi-gag-PR and env regions.

Moloney murine leukemia virus (M-MuLV) is capable of inducing promonocytic leukemia in 50% of adult BALB/c mice that have received peritoneal injections of pristane, but Friend MuLV strain 57 (F-MuLV) is nonleukemogenic under similar conditions. It was shown earlier that these differences could not be mapped to the U3 region of the virus long terminal repeat, indicating the probable influence of structural genes and/or R-U5 sequences. In this study, reciprocal chimeras containing exchanged structural genes and R-U5 sequences from these two closely related viruses were analyzed for differences in ability to induce disease. Results showed that two regions of F-MuLV, psi-gag-PR and env, when substituted for those of M-MuLV were dramatically disease attenuating. The 5'-most region, which is widely distributed, overlaps with the 5' end of the env intron and includes the RNA packaging region, psi, the entire gag coding region, and the viral protease coding region (PR) of pol. It was also found that reciprocal constructs having substitutions of both of these regions of M-MuLV in an F-MuLV background allowed full reestablishment of promonocytic leukemia. These leukemias were positive for c-myb rearrangements which are characteristic of M-MuLV-induced promonocytic leukemias. Neither region alone, however, was sufficient to produce disease with a greater incidence than 13%. Further studies demonstrated that the inability of viruses with psi, gag, PR, or env sequences from F-MuLV to induce leukemia in this model system was not due to their inability to replicate in hematopoietic tissue, to integrate into the c-myb locus early on after infection in vivo, or to express gag-myb mRNA characteristic of M-MuLV-induced preleukemic cells and acute leukemia.     

Erythromycin pharmacokinetics in children after rectal and oral administration

Pharmacokinetics of erythromycin base was studied clinically in children not older than 14 years treated with new children dosage forms of the antibiotic i. e. 0.1 and 0.25 g enteric coated tablets and 0.06 and 0.125 g suppositories. It was noted that the new dosage forms were characterized by higher availability which was 2.5-3 times higher than that after using the erythromycin base tablets without the coating. Systematic increasing of erythromycin availability after the use of the rectal suppositories was observed with increasing of the children age. Absolute absorption in newborns, sucklings and children over 1 year amounted to 28, 36 and 54 per cent respectively.     

The Effect of Material and Roughness of the Probe Surface on Probe Characteristics

Plasma Physics Reports - In the region of the surface of the probe introduced into the plasma, there are emission flows that introduce changes in the determination of its parameters. Despite the...     

Repeated oral administration of capsaicin increases anxiety-like behaviours with prolonged stress-response in rats

This study was conducted to examine the psycho-emotional effects of repeated oral exposure to capsaicin, the principal active component of chili peppers. Each rat received 1 mL of 0.02% capsaicin into its oral cavity daily, and was subjected to behavioural tests following 10 daily administrations of capsaicin. Stereotypy counts and rostral grooming were significantly increased, and caudal grooming decreased, in capsaicin-treated rats during the ambulatory activity test. In elevated plus maze test, not only the time spent in open arms but also the percent arm entry into open arms was reduced in capsaicin-treated rats compared with control rats. In forced swim test, although swimming duration was decreased, struggling increased in the capsaicin group, immobility duration did not differ between the groups. Repeated oral capsaicin did not affect the basal levels of plasma corticosterone; however, the stress-induced elevation of plasma corticosterone was prolonged in capsaicin treated rats. Oral capsaicin exposure significantly increased c-Fos expression not only in the nucleus tractus of solitarius but also in the paraventricular nucleus. Results suggest that repeated oral exposure to capsaicin increases anxiety-like behaviours in rats, and dysfunction of the hypothalamic-pituitary-adrenal axis may play a role in its pathophysiology.     

Effect of weight loss and exercise on angiogenic factors in the circulation and in adipose tissue in obese subjects

Background:

Vascular growth is a prerequisite for adipose tissue (AT) development and expansion. Some AT cytokines and hormones have effects on vascular development, like vascular endothelial growth factor (VEGF‐A), angiopoietin (ANG‐1), ANG‐2 and angiopoietin‐like protein‐4 (ANGPTL‐4).

Methods:

In this study, the independent and combined effects of diet‐induced weight loss and exercise on AT gene expression and proteins levels of those angiogenic factors were investigated. Seventy‐nine obese males and females were randomized to: 1. Exercise‐only (EXO; 12‐weeks exercise without diet‐restriction), 2. Hypocaloric diet (DIO; 8‐weeks very low energy diet (VLED) + 4‐weeks weight maintenance diet) and 3. Hypocaloric diet and exercise (DEX; 8‐weeks VLED + 4‐weeks weight maintenance diet combined with exercise throughout the 12 weeks). Blood samples and fat biopsies were taken before and after the intervention.

Results:

Weight loss was 3.5 kg in the EXO group and 12.3 kg in the DIO and DEX groups. VEGF‐A protein was non‐significantly reduced in the weight loss groups. ANG‐1 protein levels were significantly reduced 22‐25% after all three interventions (P < 0.01). The ANG‐1/ANG‐2 ratio was also decreased in all three groups (P < 0.05) by 27‐38%. ANGPTL‐4 was increased in the EXO group (15%, P < 0.05) and 9% (P < 0.05) in the DIO group. VEGF‐A, ANG‐1, and ANGPTL‐4 were all expressed in human AT, but only ANGPTL‐4 was influenced by the interventions.

Conclusions:

Our data show that serum VEGF‐A, ANG‐1, ANG‐2, and ANGPTL‐4 levels are influenced by weight changes, indicating the involvement of these factors in the obese state. Moreover, it was found that weight loss generally was associated with a reduced angiogenic activity in the circulation.     

Polymorphism of spawners of the resident form of European river lamprey <Emphasis Type="Italic">Lampetra fluviatilis</Emphasis> (Petromyzontidae)

Data on polymorphism (plastic and meristic characters) are obtained for spawners of resident nonparasitic form of European river lamprey Lampetra fluviatilis from the Serebristaya River. The new data verify the diagnosis of this species and contribute to the formulation of a hypothesis on the presence of several phenotypic groups: dwarf, small-sized, and common. Analysis of plastic characters and of the number of trunk myomeres demonstrated that the values of these characters reliably depend on position of lampreys in a certain discerned group. Some aspects of biology of this species are discussed: spawning and interaction between resident nonparasitic river and migrating parasitic lampreys.     

Photopreferendum of migratory and nonmigratory larvae of European river lamprey <Emphasis Type="Italic">Lampetra fluviatilis</Emphasis>

In the light gradient installation, the larvae of European river lamprey Lampetra fluviatilis of 0+ age, migrating downstream (downstream migrants), are distributed predominantly over the zone of dusk illumination (150–1 lx) and less so over the daylight zone (2500–150 lx) and night illumination zone (<1 lx). In 0+ larvae that passed over to the sedentary lifestyle, the photopreferendum changes: they avoid the dusk and daytime illumination and prefer nighttime illumination. Such photopreferendum is retained in later larvae of lamprey (1+ and older).     

The dynamics of nitrous oxide emission from the use of mineral fertilizers in Russia

The intensity of nitrous oxide (N2O) emission was considered based on literature data on the single input of mineral N (nitrogen) fertilizers into different agricultural soil types in Russia. Ambient environmental factors exert a combined effect on the process of gaseous nitrogen formation from fertilizers applied. To reduce the uncertainty of estimates as much as possible, only experimental results obtained under conditions similar to natural were selected for the assessments. Mineral nitric fertilizers were applied to soil at a rate of 40 to 75 kg/ha and the N2O emissions were measured for approximately 140 days. Daily average emission values varied from 0.08 to 0.45% of fertilizer nitrogen. Correspondingly, 1.26 and 2.38% of fertilizer nitrogen were emitted as N2O from chernozems and soddy podzols. In 1990, the use of fertilizers in Russian agricultural practices for 53 Gg N2O-N, which equates to approximately 6.1% of global nitrous oxide emissions from nitric fertilizers. Later, the emission dropped because of a decrease in the input of nitric fertilizers to agricultural crops, and in 1998, it constituted just 20.5% of the 1990 level. In the period from 2008 to 2012, the nitrous oxide emission is expected to vary from 0.5 to 65.0 Gg N2O-N due to possible changes in national agricultural development. In the most likely scenario, the use of mineral fertilizers in Russia will account for approximately 34 to 40 Gg N2O-N emissions annually from 2008-2012.     

90Sr and 137Cs in higher aquatic plants of the Chernobyl nuclear plant exlusion zone]

The content of radionuclides 90Sr and 137Cs in higher aquatic plants of water objects within Chernobyl NPP exclusion zone has been analysed. Biodiversity of phytocenose was studied and species-indicators of radioactive contamination were revealed. The seasonal dynamics of radionuclide content in macrophytes was studied and the role of main aquatic plant clumps in processes of 137Cs and 90Sr distribution in abiotic component of biohydrocenose was demonstrated.