Acoustically shaped DNA-programmable materials.

Recent advances in DNA nanotechnology allow for the assembly of nanocomponents with nanoscale precision, leading to the emergence of DNA-based material fabrication approaches. Yet, transferring these nano- and micron-scale structural arrangements to the macroscale morphologies remains a challenge, which limits the development of materials and devices based on DNA nanotechnology. Here, we demonstrate a materials fabrication approach that combines DNA-programmable assembly with actively driven processes controlled by acoustic fields. This combination provides a prescribed nanoscale order, as dictated by equilibrium assembly through DNA-encoded interactions, and field-shaped macroscale morphology, as regulated by out-of-equilibrium materials formation through specific acoustic stimulation. Using optical and electron microscopy imaging and x-ray scattering, we further revealed the nucleation processes, domain fusion, and crystal growth under different acoustically stimulated conditions. The developed approach provides a pathway for the fabrication of complexly shaped macroscale morphologies for DNA-programmable nanomaterials by controlling spatiotemporal characteristics of the acoustic fields.

[Immunogenicity of recombinant analog of antitumor protein lactaptin].

Therapeutic monoclonal antibodies and recombinant proteins including cytokines are commonly used in the treatment of cancer and inflammatory diseases. In most cases, these protein-based drugs exhibit a high therapeutic efficacy, which is unfortunately frequently associated with a variety of side effects. We have investigated the in vitro and in vivo immunogenicity of recombinant antitumor protein lactaptin (RL2). Based on the qRT-PCR analysis, we have shown that, in MDA-MB-231 human breast adenocarcinoma cells, RL2 suppresses the NF-kB signaling cascade that regulates the reactions of innate immunity. RL2 inhibits the expression of the CXCL1 protein and apoptosis inhibitor A20 and enhances expression of IkB, NF-kB repressor. The ELISA method has been used to evaluate the antibody titer in the blood of mice, which received single and triple intravenous or intraperitoneal injections of RL2. The multiplex immunoassay of 23 cytokines in the mice blood has shown that the RL2 injections lead to a slight increase in the levels of systemic pro-inflammatory cytokine interleukin-5 (IL-5) and keratinocyte chemoattractant (KC), a homologue of human macrophage inflammatory protein-1 (MIP-1). These observations indicate the low immunogenicity of the recombinant lactaptin analog, which can be considered to be a potential molecular drug candidate for further clinical development.

[Hygienic assessment of large-size hole type storages for low-to-moderate activity radioactive waste].

Analysis of the design features of radioactive waste (RAW) storages of large-size hole type, geological and hydrogeological conditions of their location, a multi-barrier system for environmental protection, and the reliability of a computer-aided geomonitoring system allows one assess these facilities as meeting the current International Atomic Energy Agency recommendations for RAW storages.

[Characteristics of humoral immunity to rubella virus in residents of Moscow in 1998-1999]. / Pokazateli gumoral'nogo immuniteta k virusu krasnukhi u zhitelei Moskvy v 1998-1999gg.

A total of 400 blood samples were collected from residents of Moscow in 1998-1999, 369 from adults aged mainly 19-31 years and from children aged 5-12 years. The mean incidence of antirubella antibody was 76.5%; the value varied in different age groups. The highest percentage of antibody detection was observed in men aged 19-25 years (91.8%) and the lowest in pregnant women aged 18-38 years (66-67%). Estimation of antihemagglutinin titers in international Units showed that antibody titer 1:40 and higher protected from infection; such titers were detected in 73.6% examinees. In pregnant women the level of immunological defense was very low (only 56%), which necessitates urgent vaccination of adolescent girls and young women. The results of EIA were compatible with the results of indirect hemagglutination test (IHAT), but EIA was much more sensitive in cases with low titers of IHAT.

[Virus-like particles as a source for obtaining antigens for diagnosing rubella]. / Virusopodobnye chastitsy kak istochnikh polucheniia antigenov dlia diagnostiki krasnukhi.

Rubella diagnostic agents for hemagglutination inhibition (HI) and enzyme immunoassay (EIA) based on rubella virus-like particles (RVLP) have been developed. Noninfectious RVLPs containing three structural E1, E2, and C proteins were expressed in transfected CHO24S cell culture. HI titer in culture medium was 1:256. Tween-80 treatment and ether increased HI titer 4-6-fold and rendered the antigen higher stability. Immunogenic properties of RVLPs were similar to the native rubella virus in HI test with international reference human rubella serum and sera from convalescents after rubella. Serum of mice immunized with RVLP reacted similarly with RVLP antigens and native virus. Antigen for EIA from RVLP was prepared by concentrating RVLP from culture fluid and partial purification by ultracentrifugation. The results of human sera testing by HI and EIA with RVLP and native virus antigens coincided. RVLP are identical to antigenic structure of the virus, are stable and easily purified, and can therefore be used for commercial production of HI and EIA antigens.

Effect of bilirubin on the activity and thermokinetic characteristics of brain (synaptosomal) membrane NO synthase.

NO synthase activity was found in the plasma (synaptosomal) membrane particles isolated from the homogenate of adult rat brain (without cerebellum) under conditions preventing the protease attack and formation of reactive oxygen species. The NO synthase discovered exhibited some properties of a neuronal constitutive integral membrane enzyme and was inhibited by N-nitro-L-arginine. NO synthase activity decreased when bilirubin entered the synaptosomal membrane in vitro. Bilirubin caused the shift of the transition temperature in the temperature dependence of NO synthase activity in Arrhenius plots. The incorporation of bilirubin into synaptosomal membranes resulted in an increase in the apparent activation energy for NO synthase within a temperature range of 10-30 degrees C. The membrane NO synthase was susceptible to the photodynamic effect of membrane-bound bilirubin molecules. Monomeric human serum albumin without organophilic ligands exerted a protective effect on NO synthase in bilirubin-containing membrane particles.

Stress propagation through frictionless granular material.

We examine the network of forces to be expected in a static assembly of hard, frictionless spherical beads of random sizes, such as a colloidal glass. Such an assembly is minimally connected: the ratio of constraint equations to contact forces approaches unity for a large assembly. However, the bead positions in a finite subregion of the assembly are underdetermined. Thus to maintain equilibrium, half of the exterior contact forces are determined by the other half. We argue that the transmission of force may be regarded as unidirectional, in contrast to the transmission of force in an elastic material. Specializing to sequentially deposited beads, we show that forces on a given buried bead can be uniquely specified in terms of forces involving more recently added beads. We derive equations for the transmission of stress averaged over scales much larger than a single bead. This derivation requires the ansatz that statistical fluctuations of the forces are independent of fluctuations of the contact geometry. Under this ansatz, the d(d+1)/2-component stress field can be expressed in terms of a d-component vector field. The procedure may be generalized to nonsequential packings. In two dimensions, the stress propagates according to a wave equation, as postulated in recent work elsewhere. We demonstrate similar wave-like propagation in higher dimensions, assuming that the packing geometry has uniaxial symmetry. In macroscopic granular materials we argue that our approach may be useful even though grains have friction and are not packed sequentially.

[The effect of cobalt chloride on hemoxygenase activity, on bilirubin metabolic homeostasis in the brain and on serum albumin transport function in animals]. / Vliianie khlorida kobal'ta na aktivnost' gemoksigenazy, metabolicheskii gomeostaz bilirubina v golovnom mozge i transportnuiu funktsiiu syvorotochnogo al'bumina u zhivotnykh.

Administration of cobalt chloride (everyday dosage 50-100 mumol/kg) to rats in the course of pregnancy and first two days of postnatal period increases the activity of hemoxygenase + NADPH biliverdinreductase enzyme system in the brain of adult rats and induce this activity in new-born rats. Functional activity of serum albumin connected with bilirubin transport, the pH value in blood and bilirubin concentration in the brain of adult and new-born rats virtually do not change, whereas bilirubin concentration in the serum of pregnant rats received CoCl2 raises. When the same doses of cobalt chloride are administered to pregnant rats in combination with the mixture of high fatty acids and acetylsalicylic acid the activity of this enzyme system in the brain increases, functional activity of serum albumin connected with bilirubin transport decreases, bilirubin concentration in the brain raised (serum bilirubin concentration being comparatively low) at the same pH value in blood of both pregnant and new-born rats.

[Interaction of human serum albumin with bilirubin in solution and in suspensions containing bile pigment of membranous particles]. / Vzaimodeistvie syvorotochnogo al'bumina cheloveka s bilirubinom v rastvore i v suspenzii soderzhashchikh zhelchnyi pigment membrannykh chastits.

Spectrophotometrically measured in homogeneous aqueous medium association constants for the formation of bilirubin--human human serum albumin complex did not correlate with the intensity of the same complex formation as a result of bilidiene release from nerve cell plasma membrane particles containing biliary pigment in aqueous medium during changed hydrogen parameter and ionic strength of the medium (pH22 degrees 6.8, 7.2, or 7.5 and NaCl concentration in the medium 0.135, 0.150, or 0.165 M), such changes being compatible with normal mammalian cellular and tissue vital activity, the rest conditions being compatible. Study of the time course of the said complex formation in membranous particle suspension seems to be more adequate and therefore more reliable for organic anions (e.g. bilirubin) transport related assessment of detoxifying activity of infused human serum albumin preparations than measurement of the said association constants in a homogeneous aqueous medium.

[Disorder of the serum albumin transport function in chronic oxygen starvation and its pharmacological correction]. / Narushenie transportnoi funktsii syvorotochnogo al'bumina pri khronicheskom kislorodnom golodanii i ee farmakologicheskaia korrektsiia.

Grade II chronic intermittent hypoxic hypoxia under normal barometric pressure, which was reproduced in pregnant rats 2.0-2.5 weeks before expected delivery and in the offsprings in the first 2 days of neonatality caused a decrease in serum albumin functional activity associated with bilirubin transport, as evidenced in vitro, as well as accumulation of serum albumin-bound nonesterified higher fatty acids and acetaldehyde along with uncompensated acidosis, an increase in brain levels of bilirubin in adult and newborn rats, and enhancement of its hemoxygenase activity. Administration of alpha-tocopherol acetate, nicotinamide, sodium oxybutyrate, and trisamine to animals exposed to hypoxia normalized the above biochemical parameters, except for hemoxygenase activity and promoted a greater binding of endogenous bilirubin to albumin in adult and newborn rats in vivo.

[Possible reasons for bilirubin accumulation in the brain of newborn animals during oxygen deficiency]. / Vozmozhnye pricheny nakopleniia bilirubina v golovnom mozgu novorozhdennykh zhivotnykh pri kislorodnoi nedostatochnosti.

An increase in content of bilirubin was detected in brain tissue of newborn rats with chronic interrupted hypoxic hypoxia occurred during pre- and neonatal periods. Accumulation of bilidiene in animal brain tissues appears to occur due to the following causative factors: tissue acidosis, developed after oxygen deficiency, contributed to binding of bile pigment to brain cell plasma membranes, synaptosomal ones in particular; serum albumin lost its specific ability to bind bilirubin due to protein molecular accumulation of excessive fatty acids and acetaldehyde, which are bound to protein, as well as other minor ligands extracted by organic solvents.

[Coding part of the son gene small transcript contains four areas of complete tandem repeats]. / Kodiruiushchaia chast' posledovatel'nosti malogo transkripta gena son soderzhit chetyre oblasti sovershennykh tandemnykh povtorov.

Overlapping cDNA clones for the smaller son gene transcript were isolated by several steps of human placenta cDNA library walking. Here we report the structure of repeats that exist at the long open reading, present in the structure of this smaller (5.6 kb) transcript of human gene son. We were able to identify in this transcript four areas of complete tandem repeats, formed by several steps of duplication. For three of them the extent of homology between amino acid sequences of repeated elements is higher than between their nucleic acid sequences. It means that amino acid sequences of repeated elements are conserved, that could reflect the structural significance for the function of hypothetical protein product of human gene son. On the basis of the results of analysis we proposed the model for the formation of repeats by processes of duplication, mutation and natural selection pressure at the expressed protein level.

Exons I and VII of the gene (Ker10) encoding human keratin 10 undergo structural rearrangements within repeats.

A genomic fragment containing the K51 gene previously isolated from a rat genomic library by hybridization with the v-mos probe in nonstringent conditions [Chumakov et al., Dokl. Akad. Nauk SSSR 290 (1986) 1252-1254], resembles a human keratin type-I-encoding gene [Shvets et al., Mol. Biol. 24 (1990) 663-677]. This genomic clone, K51, has been used as a probe to search for related human genes. A recombinant clone, HK51, with a 1.5-kb insert, was isolated from a human embryonic skin cDNA library, and its nucleotide (nt) sequence was determined. Analysis has shown that the cloned cDNA encodes human keratin 10 (Ker10). All presently known nt sequences of the human Ker10-encoding gene (Ker10) are not identical. Differences are concentrated in the 5'-end of the first exon and in the middle of the seventh exon within repeats. In spite of structural rearrangements in two of eight exons, the reading frame and position of the stop codon are preserved. The genetic rearrangements cause changes in hydrophobicity profiles of the N and C termini of Ker10. It was also noticed that insertion of one nt leads to the formation of an unusual 3'-end of the transcript.

The expression of skin-specific gene K51 in the epidermal layer of human skin and in basal cell carcinoma cells.

Gene K51 probe isolated previously from the rat genomic library has been used to study the expression of its human counterpart by in situ hybridization and Northern blot analysis. A polyA-containing transcript of human gene K51 of 3 kb size has been detected in embryonic skin. The gene is also expressed in the epidermis of newborn humans and adults, but not in the adjacent mesenchymal tissues. Immunostaining with keratin antisera revealed predominantly earlier stage expression of K51 than cytokeratin markers. Sebaceous and sweat glands also contain cells expressing K51 gene. K51 expression was found in the cells of eight individual basal cell carcinomas tested, with the level of expression lower than in keratinocytes from normal human epidermis. We propose that K51 gene expression could serve as a convenient marker for the study of the process of skin keratinocyte development and the changes in this process associated with skin cancers and dysplasia.