Enhancement of growth promoting activity of human blood on keratinocytes after its irradiation in vivo (transcutaneously) and in vitro with visible and infrared polarized light

To stimulate wound healing, current medicine uses various methods of phototherapy. The induced activation of proliferative processes in the wound occurs due to development of not only local, but also systemic processes, whose nature remains largely uninvestigated. The present work provides evidences that as early as 30 min after irradiation of a small area of the volunteer's body surface with polychromatic visible light + infrared polarized light (400-3400 nm, 95% of polarization) at a therapeutic dose (12 J/cm2), soluble factors appear in the circulating blood, which are able to stimulate proliferation of human keratinocytes in primary culture. A similar effect was also revealed after a direct blood irradiation. A proof is provided in favor of a hypothesis that a rapid rise of growth promoting activity of the entire circulating blood may be a consequence of transcutaneous photomodification of the small amount of light-modified blood in superficial skin vessels, and of the effect of such blood on its entire circulating volume. A possibility of a release into plasma of growth factors from blood cells and complexes with alpha 2-macroglobulin is discussed.     

Autoradiographic study of RNA synthesis and secretion into the cytoplasm during the early stages of liver regeneration

Autoradiographic experiments with regeneration of rat liver showed that RNA transport from nucleus to cytoplasm occurs more quickly in 3h following partial hepatectomy while the total RNA synthesis increases insignificantly. Nucleolar RNA synthesis at this time grows in some hepatocytes and decreases in others though it does not differ on the average from the control one. By the 6th h after partial hepatectomy total RNA synthesis significantly increases simultaneously with the acceleration of RNA transport to cytoplasm. Nucleolar RNA synthesis intensifies in all hepatocytes.     

Similarities in Restriction Fragment Patterns of Mitochondrial DNAs of PhytophthoraSpecies Strongly Depend on the Restriction Enzyme Used Due to Heterogeneous Base Distribution and Sequence Conservation

Mitochondrial DNAs of six morphologically different Phytophthora species were digested with 15 restriction enzymes. The numbers of restriction fragments obtained differed considerably from those theoretically expected for random base distribution. Enzymes with relatively many G and C in their recognition sequences produced significantly larger numbers of fragments. Moreover, fragments generated by most of these enzymes were more often shared by two or more species than those from enzymes with more A and T in their recognition sequence. It is concluded that base distribution in mitochondrial DNA of Phytophthora is heterogeneous,AT-rich stretches occurring scattered over the mitochondrial genome and GC-rich regions present in conserved sequences, presumably genes. A practical consequence for taxonomic RFLP studies is that optimal enzymes can be selected, depending on the desired level of resolution.     

Tetrapod phylogeny inferred from 18S and 28S ribosomal RNA sequences and a review of the evidence for amniote relationships [published erratum appears in Mol Biol Evol 1991 May;8(3):398]

The 18S ribosomal RNAs of 21 tetrapods were sequenced and aligned with five published tetrapod sequences. When the coelacanth was used as an outgroup, Lissamphibia (living amphibians) and Amniota (amniotes) were found to be statistically significant monophyletic groups. Although little resolution was obtained among the lissamphibian taxa, the amniote sequences support a sister-group relationship between birds and mammals. Portions of the 28S ribosomal RNA (rRNA) molecule in 11 tetrapods also were sequenced, although the phylogenetic results were inconclusive. In contrast to previous studies, deletion or down- weighting of base-paired sites were found to have little effect on phylogenetic relationships. Molecular evidence for amniote relationships is reviewed, showing that three genes (beta-hemoglobin, myoglobin, and 18S rRNA) unambiguously support a bird-mammal relationship, compared with one gene (histone H2B) that favors a bird- crocodilian clade. Separate analyses of four other genes (alpha- crystallin A, alpha-hemoglobin, insulin, and 28S rRNA) and a combined analysis of all sequence data are inconclusive, in that different groups are defined in different analyses and none are strongly supported. It is suggested that until sequences become available from a broader array of taxa, the molecular evidence is best evaluated at the level of individual genes, with emphasis placed on those studies with the greatest number of taxa and sites. When this is done, a bird-mammal relationship is most strongly supported. When regarded in combination with the morphological evidence for this association, it must be considered at least as plausible as a bird-crocodilian relationship.      

Enhancement of fibroblast growth promoting activity of human blood after its irradiation in vivo (transcutaneously) and in vitro with visible and infrared polarized light

Visible and infrared (IR) irradiation of laser and non-laser sources has a pronounced wound-healing effect promoting tissue repair without hyperproduction of connective tissue elements. This effect develops as a consequence of local and systemic light effects, but many aspects of their mechanism have been yet unclear. In the present work, we have shown that in 0.5 h after irradiation of a small area of the volunteers' body surface with polychromatic visible + IR light (400-3400 nm, 95% polarization, 12 J/cm2) the amounts of PDGF and TGF-beta 1 in the blood serum increase, on average, by 20 and 43%, respectively. This effect is preserved for at least 24 h to be recorded only in volunteers with the initially normal and decreased levels of the growth factors; the initially elevated content of PDGF-AB decreases. Addition of such a plasma (2.5%) to the nutrient medium of primary cultures of human embryonal fibroblasts stimulates cell proliferation, on average, by 10 and 17%, but only in the case if the initial growth-promoting (GP) blood activity was low. Similar changes occur in parallel experiments following irradiation of blood samples of the same volunteers in vitro, as well as at mixing irradiated and non-irradiated autologous blood at the ratio 1:10 (v/v), i.e. at modeling a situation in the vascular bed, when the transcutaneously photomodified blood contacts with the rest of its volume. Similar changes in the blood GP activity under conditions in vitro were recorded as well after 4-9 daily phototherapy sessions. This allows us to suggest that changes in GP activity of circulating blood of the irradiated volunteers may be, to a large extent, the consequence of effect exerted on the blood by small amounts of transcutaneously photomodified blood. The obtained results are discussed in terms of light effect on wound healing and scar tissue formation, with regard to the authors' previous data on much higher GP of the irradiated blood in respect to keratinocytes, the fast decrease in proinflammatory cytokine levels, and the increase in IFN-gamma content.     

Changes in cytokine content in the peripheral blood of volunteers after their exposure to polychromatic visible and infrared light

Anti-inflammatory, immunomodulating and wound-healing effects of visible and infrared (IR) radiation from laser and non-laser sources are widely used in current medicine. However, the role of pro- and anti-inflammatory cytokines in development of these effects has been poorly studied. A randomized, placebo-controlled, double blind study was made. Using ELISA, the content of 10 cytokines was studied in the peripheral blood of volunteers after a single and four daily irradiations of the sacral area (D = 15 cm) with polychromatic visible + IR polarized light (480-3400 nm, 12 J/cm2). The phototherapeutic sessions were accompanied by four blood exfusions for the study (to a total volume of 80 ml). In the control (placebo) group, irradiation was imitated, and blood samples of the same volume were drawn at the same time intervals as in volunteers of the main group. A fast decrease in the level of pro-inflammatory cytokines was revealed as soon as in 0.5 h after the irradiation. This level was retained until the end of the phototherapeutic course. At the parameters exceeding the norm, the contents of TNF-alpha, IL-6 and IFN-gamma fell, on average, by 34, 12 and 1.5 times, respectively. By the end of the course, the levels of IFN-gamma and of IL-12 decreased by 5 and 15 times, respectively. A fast decrease (by two-fold) was also characteristic of normal values of IL-6. Neither IL-1beta, nor IL-2 were detected in blood plasma of the examined people both before and after the irradiation. In parallel with a decrease in the proinflammatory factor levels the amount of anti-inflammatory cytokines was found to rise: that of IL-10--by 2.7-3.5 times in 0.5 h and at later terms at the initially normal parameters, and that of TGF-beta1--by 1.4-1.5 times at the initially decreased level. The IL-4 content did not change. A characteristic feature of the light effect was a fast rise of IFN-gamma amount--by 3.3-4.0 times in individuals with its initially normal level, with no changes in IFN-alpha content. The above-reported regularities of the light effects were also recorded at a direct (in vitro) irradiation of the examined volunteers' blood, as well as on addition of irradiated blood to a 10-fold volume of non-irradiated autologous blood, i.e. at a modeling of mixing, of a small amount of transcutaneously photomodified blood with its main circulating volume in the vascular bed of an irradiated person. Such a similarity of effects in blood following its irradiation in vivo and in vitro enables us to associate the fast changes of the cytokine content in the entire volume of peripheral blood with the transcutaneous photomodification of its small amounts, and with a "transfer" of the light effects by photomodified blood to the whole pool of circulating blood.     

Changes in the expression of membrane markers and in the number of human blood monocytes after single and repeated courses of visible and infrared light at therapeutic doses

An attempt has been made to prove that the immunomodulating effect of therapeutic doses of polychromatic visible + infrared polarized (VIP) light at its application to a small body surface area is connected with a transcutaneous photomodification of a small amount of blood in superficial skin microvessels. For this purpose, in parallel experiments, using monoclonal antibodies, the membrane phenotype of circulating blood mononuclears was studied after irradiation of volunteers, of samples of their blood in vitvo, and of a mixture of the irradiated and non-irradiated autologous blood in a 1:10 volume ratio, thereby modeling events in vivo, when a small amount of the transcutaneously photomodified blood in the vascular bed contacts its main circulating volume. In this variant of experiment, a great similarity has been established of changes in expression of mononuclear membrane markers (CD3, CD4, CD8, CD20, CD16, HLA-DR and to a lesser degree of CD25); the ability has been proven of the photomodified blood to "translate" the light-induced changes to a much higher volume of non-irradiated blood, which might represent a mechanism of the systemic immunomodulating effect of phototherapy. Under conditions in vivo and in vitro, the most "reactive" were HLA-DR+, CD20+, CD16+, CD4+, and 0-cells. An increase of the total number of lymphocytes and monocytes has been shown by the end of the 10-day-long phototherapeutic course. The regulatory character of the single and course sessions of the VIP light on the blood immunocompetent cells is substantiated: depending on the initial state of the immune system, the VIP light can produce both stimulating and inhibitory effect on lymphoid cell subpopulations, which opens large possibilities of using this method for correction of immunological disturbances in diseases of different etiopathogenesis.