Registration of surface fluorescence of endogenous porphyrins during low-level laser therapy of gastric and duodenal ulcers

With the use of the developed opto-fiber spectroscope and gastroscope, the intensity of fluorescence from the ulcer surface into the gastrointestinal tract was measured in the range of 670-690 nm under the He-Ne laser (= 632.8 nm) illumination. Surface fluorescence data were collected with the use of a special diagnostic fiber with a total diameter of about 2 mm, which combines some illuminating and receiving fibers. To do the measurements, the diagnostic fiber was brought into a slight contact with the mucosa surface. The quantitative results were calculated with the use of the special fluorescent contrast coefficient (Kf), which takes into account the backscattered light as well. After the fluorescent diagnostic procedure, the low-level laser therapy was applied to all patients. It was found that: first of all, during the laser medical cure the registered fluorescence for the patients with a good dynamics of treatment had an evident tendency to a decrease, and the coefficient Kf for observed ulcers tended to approach the value obtained from normal mucosa (Kf), which indicates the normalization of the porphyrin content in the tissue. Second, the retrospective analysis of the efficacy of laser therapy versus initial values of Kf showed that, for the patients with the initial values Kf = 0.2, the efficiency of the laser treatment was the highest: the relation between the patients with good treatment results and bad ones was 4:1 (more than 80%). For the initial Kf = 0.2-0.3, this relation was 3:1 but for Kf = 0.4-0.5 it was 2:3. The differences in the efficiency of the low-level laser therapy, except the cases of the cancer in the ulcers, may be due to different content of porphyrin in the tissue: at high concentrations of the photosensitizer in the ulcer, the effect of the overdosing could be realized. In this case, another technology of laser illumination is needed. The results are discussed in terms of the free-radical conception of the stimulating effect of laser radiation.     

Effect of low-level laser therapy on experimental wounds of hard palate mucosa in mice

Under general anesthesia and sterile conditions, incision wound was induced in the hard palate mucosa of adult male mice. The wounds of groups 1 and 2 were irradiated daily with He-Ne laser at 3 and 7.5 J/cm2 for 120 and 300 s, respectively, while the incision wound of group 3 not exposed served as controls. On day 3 of injury, the laser-treated wounds contained significantly lower neutrophils than the wounds in the control group. By day 7 after injury, the laser-treated wounds contained significantly more fibroblasts and at the same time contained significantly fewer macrophages. In conclusion, an acceleration of the wound healing process of experimental wounds in the hard palate mucosa of mice at low-level laser therapy with a He-Ne laser at energy densities of 3 and 7.5 J/cm2 was observed.     

The effect of low-intensity laser irradiation in the blue, green, and red spectral ranges on healing of experimental skin wounds in rats

The effects of low-intensity laser irradiation in the red (632.8 nm), green (532 nm), and blue (441.2 nm) spectral ranges on wound healing has been studied in rats. The effect of the traditionally used red laser irradiation has been compared with the effect caused by laser irradiation in other spectral ranges, aiming to support the provisional hypothesis that a similar healing effect could be achieved at lower doses of wound irradiation by lasers emitting in the blue and green spectral ranges. The following parameters have been used to characterize healing of the experimental wounds: the functional activity of phagocytes in the wound exudate, which was determined from luminol-dependent chemiluminescence, the phagocyte number; the wound exudates’ antioxidant activity; and the rate of healing, which was determined as the change of the wound surface area. It was found that in all cases the laser irradiation accelerated the healing of wounds. Exposure to red laser irradiation at the dose of 1.5 J/cm²), and to blue or green laser irradiation at a dose of 0.75 J/cm² shortened the time of the wound healing from 22 to 17 and 19 days, respectively. The functional activity of leukocytes in irradiated groups increased by day 5 after surgery, whereas in the control group it decreased. The superoxide dismutase activity increased in all experimental groups by day 5 after surgery. Laser irradiation in the red spectral range at a dose of 1.5 J/cm² resulted in a larger increase in superoxide dismutase activity, as compared to that found after exposure to laser irradiation in the blue and green spectral ranges at a dose of 0.75 J/cm².     

Dynamics of wound healing of skin wounds produced by laser and scalpel in experiments

In this study in 90 rats the measured rate of healing, pO2 in sites of postoperative scar and histological specimen of identical excisional wounds produced by CO2-laser, scalpel and combination are compared. The data of healing laser wounds via scalpel ones show slow laser wound healing. We explain, this by inhibition of the inflammation phase due to diminishing macrophages migration into the wound. pO2 data in postoperative scar after laser and scalpel wounds show that laser thermal alteration does not influence CO2 regimen in the scar.     

Role of endogenous porphyrins in the effects of low-intensity laser radiation of the red region on free radical processes in the blood of rats under experimental endotoxic shock

The role of endogenous porphyrins in the effects of laser radiation of the red region (632.8 nm) on free radical processes in the blood of rats under endotoxic shock induced by the administration of lipopolysaccharide B (25 mg/kg) has been studied. The measurements of the functional activity of polymorphonuclear leukocytes (the method of luminol-dependent chemiluminescence), the superoxide dismutase activity of blood plasma (using nitro blue tetrazolium), and the degree of lipid oxidation of erythrocyte membranes (the method of fluorescence of cis-parinaric acid) have been carried out. It has been found that low-intensity laser radiation strongly affects all processes examined irrespective of the administration of lipopolysaccharide B. The effect of radiation was most pronounced in animals injected with the polysaccharide, the changes being dependent on the concentration of endogenous porphyrins in samples.     

Role of endogenous porphyrins in the effects of low-intensity laser radiation of the red region on free radical processes in the blood of rats under experimental endotoxic shock

The role of endogenous porphyrins in the effects of laser radiation of the red region (632.8 nm) on free radical processes in the blood of rats under endotoxic shock induced by the administration of lipopolysaccharide B (25 mg/kg) has been studied. The measurements of the functional activity of polymorphonuclear leukocytes (the method of luminol-dependent chemiluminescence), the superoxide dismutase activity of blood plasma (using nitro blue tetrazolium), and the degree of lipid oxidation of erythrocyte membranes (the method of fluorescence of cis-parinaric acid) have been carried out. It has been found that low-intensity laser radiation strongly affects all processes examined irrespective of the administration of lipopolysaccha-ride B. The effect of radiation was most pronounced in animals injected with the polysaccharide, the changes being dependent on the concentration of endogenous porphyrins in samples.     

Healing of abdominal skin wounds in rats

The studies performed on rats have shown that healing of square full-tissue skin wounds depends on their location. Thus, the healing of wounds on the belly was faster and more complete than that of dorsally located ones. Regeneration of belly skin resulted in the formation of new tissue with a typical network of fibers and skin folds. On the back epithelial connective tissue scars were formed.     

The role of endogenous thiols in intrinsic radioprotection

Observations are reviewed from experiments performed to study the role of endogenous thiols in the radiation response of cells using a glutathione-deficient and a related glutathione-proficient cell strain. The effect of glutathione in the initial radical reactions was considered and the yield of single-strand DNA breaks was the end-point of the response. The rejoining of breaks and clonogenic survival were chosen as end-points when, in addition, the role of glutathione in the subsequent biochemical processes was studied. The results were interpreted to indicate that glutathione plays a role in both the radical and the biochemical reactions which follow irradiation. In the former case, it functions as a damage-restituting reactant, in general agreement with the 'competition model'. Some biochemical repair processes, in particular those concerned with the rejoining of breaks induced by radiation in the presence of oxygen or misonidazole, appear also to be critically dependent on glutathione. Due, probably, to its particular spatial distribution, endogenous glutathione is specific in the radical processes, and exogenous thiols cannot be substituted for it. No such specificity was indicated in the biochemical processes related to strand break rejoining.     

The role of endogenous auxin in root initiation

This paper describes the process of the formation of adventitious roots. There appears to be good agreement that this consists of four stages, defifferentiation coupled with the formation of a meristematic locus, cell division to form a radially symmetrical cluster of cells, further divisions coupled with organisation into a bilaterally symmetrical meristem and finally growth of cells in the basal part of the meristem which causes its protursion through the epidermis. Evidence for the involvement of auxins in these various stages is reviewed and the extent to which rooting of easy- and hard-to-root species can be accounted for in terms of auxin content discussed. Peaks of IAA occur soon after excision of cuttings in some species and there is some evidence suggesting that this is correlated with changes in peroxidase activity. The possible involvement of cytokinins with auxins is briefly considered.     

The role of endogenous auxin in root initiation

This paper is the second part of a review which considers evidence for the involvement of auxin in root initiation. Part II examines the research being carried out with transformed plant tissues. Agrobacterium rhizogenes causes abundant root initiation at the site of inoculation. Ri plasmid T-DNA contains several genes which encode enzymes involved in the biosynthesis and metabolism of indole-3-acetic acid. Transfer of various fragments of the Ri plasmid has also been reported to confer increased sensitivity to auxin upon plant cells. Controlled expression of these genes in the plant genome potentially offer an insight for developmental plant physiologists into the role of plant growth substances in the process of root initiation. The importance of absolute levels of IAA in the stimulation of root initiation is discussed.     

Reinnervation of experimental superficial wounds in rats

Sensory reinnervation of a superficial skin wound in the rat was studied by labeling sensory axons with anterogradely transported wheat germ agglutinin-horseradish peroxidase. Reinnervation starts after 3 days from the edge of the wound as well as from beneath the wound. About 2 weeks after the production of the wound, some hyperinnervation appears to be present, but after a few additional weeks, the innervation pattern is essentially normal. The results indicate that structural recovery of sensory axons is rapid and probably complete when skin wounds heal with no or minimal scar formation.     

Interactions with glutathione S-transferases of porphyrins used in photodynamic therapy and naturally occurring porphyrins.

Several naturally occurring porphyrins and porphyrins used in photodynamic therapy inhibit glutathione S-transferase isoenzymes either purified from rat liver or lung or in cytosol from normal and from cancerous (Morris 7288C hepatoma) liver. Although differences occur in the type and amount of transferases in normal and cancerous liver and in the liver of rats bearing an extrahepatic tumour, these enzymes are potential binding sites for porphyrins. Porphyrin structure is an important factor in determining the affinity of binding, as shown by the relative inhibitory effectiveness. Of the dicarboxylic porphyrins in the mixture used clinically, OO'-diacetylhaematoporphyrin and monohydroxyethylmonovinyldeuteroporphyrin are more effective inhibitors than haematoporphyrin and protoporphyrin IX. Of the naturally occurring porphyrins the order of effectiveness is protoporphyrin IX (dicarboxylic) greater than coproporphyrin (tetracarboxylic) greater than uroporphyrin (octacarboxylic) and type I greater than type III isomers of both uroporphyrin and coproporphyrin, and the synthetic tetra-meso-phenylporphinetetrasulphonate is a better inhibitor (apparent Ki = 250 nM) than coproporphyrin, which contains a comparable number of negative charges. In addition, iron-porphyrin chelates are more effective inhibitors of the transferases, with 25-fold decrease in Ki value, than the free porphyrins. These results indicate that one means whereby porphyrins accumulate in tissues is the occupation of intracellular binding sites, such as the transferases. Since porphyrins inhibit the activity of these important detoxifying enzymes, there will be metabolic consequences to the cell.     

The role of endogenous bromotyrosine in health and disease

Abstract

Bromotyrosine is a stable by-product of eosinophil peroxidase activity, a result of eosinophil activation during an inflammatory immune response. The elevated presence of bromotyrosine in tissue, blood, and urine in medical conditions involving eosinophil activation has highlighted the potential role of bromotyrosine as a medical biomarker. This is highly beneficial in a paediatric setting as a urinary noninvasive biomarker. However, bromotyrosine and its derivatives may exert biological effects, such as protective effects in the brain and pathogenic effects in the thyroid. Understanding these pathways may yield therapeutic advancements in medicine. In this review, we summarize the existing evidence present in literature relating to bromotyrosine formation and metabolism, identify the biological actions of bromotyrosine and evaluate the feasibility of bromotyrosine as a medical biomarker.     

The role of carboxymethyl substituents in the interaction of tetracationic porphyrins with DNA

Cationic porphyrin-based compounds capable of interacting with DNA are currently under extensive investigation as prospective anticancer and anti-infective drugs. One of the approaches to enhancing the DNA-binding affinity of these ligands is chemical modification of functional groups of the porphyrin macrocycle. We analyzed the interaction with DNA of novel derivatives containing carboxymethyl and ethoxycarbonylmethyl substituents at quaternary nitrogen atoms of pyridinium groups at the periphery of the porphyrin macrocycle. The parameters of binding of 5,10,15,20-tetrakis(N-carboxymethyl-4-pyridinium)porphyrin (P1) and 5,10,15,20-tetrakis(N-ethoxycarbonylmethyl-4-pyridinium)porphyrin (P2) to double-stranded DNA sequences of different nucleotide content were determined using optical spectroscopy. The association constant of P1 interaction with calf thymus DNA (K?=?3.4?×?10(6)?M(-1)) was greater than that of P2 (K?=?2.8?×?10(5)?M(-1)). Preferential binding of P1 to GC- rather than AT-rich oligonucleotides was detected. In contrast, P2 showed no preference for particular nucleotide content. Modes of binding of P1 and P2 to GC and AT duplexes were verified using the induced circular dichroism spectra. Molecular modeling confirmed an intercalative mode of interaction of P1 and P2 with CpG islands. The carboxyl groups of the peripheral substituent in P1 determine the specific interactions with GC-rich DNA regions, whereas ethoxycarbonylmethyl substituents disfavor binding to DNA. This study contributes to the understanding of the impact of peripheral substituents on the DNA-binding affinity of cationic porphyrins, which is important for the design of DNA-targeting drugs.     

The role of melanocortins in skin homeostasis

Melanocortins are structurally related bioactive peptides which are produced by many extra-neural tissues including the skin. All of the melanocortins (alpha, beta, and gamma-melanocyte-stimulating hormone and adrenocorticotropin) have melanotropic activity but can elicit many other effects on skin cells. On the basis of in vitro and in vivo findings melanocortins have been shown to regulate immune and inflammatory responses, hair growth, exocrine gland activity and extracellular matrix composition. These effects are mediated by melanocortin receptors among which the melanocortin-1 receptor is most ubiquitously expressed by human skin cells. Simultaneous expression of melanocortins and their receptors suggest a complex autocrine and/or paracrine regulatory network whose disruption invariably affects skin homeostasis. Expression of melanocortin receptors on various skin cell types further indicates novel pharmacological targets for the treatment of skin diseases.     

A study of the effect of low-intensity laser radiation of the blue, green, and red spectral regions on the healing of experimental skin wounds in rats

The effect of low-intensity laser radiation of the blue (441.2 nm), green (532 nm), and red (632.8 nm) spectral regions on the healing of experimental skin wounds in rats has been studied. The effect of the traditionally applied laser radiation in the red region has been compared with the effect of laser radiation in the other spectral regions, assuming that, upon irradiation of wounds by lasers emitting in the blue and green regions, a similar effect can be achieved at lower doses. The following parameters characterizing the healing of experimental wounds were used: the functional activity of phagocytes of wound exudates, which was determined by luminol-dependent chemiluminescence, and their number; the antioxidant activity of wound exudates; and the rate of healing, which was determined as a change in the wound area. It was shown that irradiation with laser accelerated the healing of wounds in all cases. The exposure to laser radiations in the red (1.5 J/cm), blue, and green (0.75 J/cm2) spectral regions shortened the time of wound healing from 22 to 17 and 19 days, respectively. The functional activity of leukocytes after the exposure increased on day 5 after the infliction of the wound, whereas in the control it decreased. The superoxide dismutase activity increased in all experimental groups by day 5 after the operation. A maximum increase in the superoxide dismutase activity occurred after the exposure to laser radiation in the red region at a dose of 1.5 J/cm and in the blue and green spectral regions at a dose of 0.75 J/cm2.     

Effect of a low-intensity CO2 laser on the process of reparative regeneration of experimental wounds

The authors studied the effect of CO2 laser with an energy density ranging from 0.1 to 300 mVt/cm2 on reparative regeneration of experimental wounds. The criteria of the efficacy of laser therapy were the wound surface area, the surface of the necrotic area, cellular composition and the content of collagen and glycosaminoglycans in the granulation tissue. Laser radiation with an energy density of 0.5 and 4 mVt/cm2 and exposure of 2 min appeared to produce the most beneficial effect on reparative regeneration.