A clinical evaluation of efficacy of photodynamic therapy in treatment of reticular oral lichen planus: A case series.

Background The aim of the study was to clinically evaluate the efficacy of photodynamic therapy in treatment of reticular oral lichen planus (OLP). Methods Fifty patients aged 26-84, with 124 OLP lesions in total, underwent photodynamic therapy (PDT) mediated with topically applied 5% 5-aminolevulinic acid. ALA was activated by a custom-made diode lamp with a high-power LED emitting light at 630 nm and 300 mW delivered through an optical fiber probe. A light exposure dose was 150 J/cm2. The therapy comprised of 10 weekly illumination sessions. The lesions' response was macroscopically measured in millimeters with a periodontal probe and clinically evaluated at each session, then on completion of the series and throughout the 12-month follow-up. Results The baseline mean size of lesions was 3.99 cm2±3.73. The lesions on the buccal mucosa and lips (lining mucosa) were larger than those on the gingiva and tongue (masticatory mucosa) - 4.58 cm2±4.01 and 2.93 cm2±2.91 respectively. On completion of the therapy 109 sites improved, including 46 in complete remission. The mean reduction in size was 62.91% (p = 0.000000). 12-month after therapy mean reduction of the lesions was 78.7% (p = 0.000000), specifically 79.48% (p = 0.000000) within the lining mucosa and 76.11% on the masticatory mucosa. Conclusions The results proved that ALA-mediated photodynamic therapy with a 630 nm light was effective and as such it can be used as an optional treatment for symptomatic OLP.

Homocysteine as a Diagnostic and Etiopathogenic Factor in Children with Autism Spectrum Disorder.

Substantial characteristics of autism are cognitive and psychophysical disorders. Etiopathogenetic factors are thought to be responsible for development of autism in children with genetic predisposition as well as have their effect on the severity of the disorders. The main problem of early identification of patients affected by autism spectrum disorder is that there are no clear diagnostic criteria. The aim of our study was assessment of hair magnesium and serum homocysteine concentrations in children with autism. The presented work is a continuation of previous study in which we investigated the influence of disturbances in magnesium and homocysteine levels in children with autism, performed on a new, larger group of patients. One hundred and forty children had hair magnesium levels analyzed, as well as blood serum levels of homocysteine and magnesium. Hair magnesium analysis was performed using a flame atomic absorption spectrometer, blood serum homocysteine determination was performed using a radioimmunological method, and blood serum magnesium level was determined using a biochemical method. Our research showed normal magnesium blood levels and significantly high homocysteine levels and very low hair magnesium levels. Low concentration of hair magnesium progresses with age. Our hypothesis is that magnesium deficiency, as a relevant epigenetic factor, might be decreasing methylation of homocysteine, therefore decreasing genome transcription and lowering the synaptic plasticity. We suggest that analysis of hair magnesium and serum homocysteine levels might be useful in identification of children with autism spectrum disorder, as well as control of its treatment. Obtained results and performed analysis might therefore justify supplementation of magnesium among children with autism.

Bioelements in hair of children with selected neurological disorders.

We have analyzed concentrations of magnesium (Mg), calcium (Ca), copper (Cu), zinc (Zn) and iron (Fe) in hair of a group of 82 children with mental retardation, in which 9 patients suffered from epilepsy, 18 from the Down's syndrome and 55 from cerebral palsy. Girls comprised little over 50% of the patients. In the group of boys with epilepsy, we found Mg, Ca, Cu and Fe deficiency, and normal level of Zn. In the group of girls with epilepsy, apart from low Fe concentration, a high level of Ca, Mg, Zn, and Cu was noted. For girls with the Down's syndrome, a high or normal level of Ca, Mg, Zn and Cu was found, whereas the Fe concentration varied and presented itself in a non-characteristic way. Both groups of children with cerebral palsy, i.e. boys and girls, displayed low Fe concentration in their hair; low Cu level was found in older patients as well. In this group of patients, we also noted high concentrations of Ca, Mg and Zn in girls and normal in boys. A high concentration of Ca in girls with cerebral palsy requires separate analysis. The obtained results could be useful as guidance in the direction and determination of the amount of possible patient nutritional supplementation.

A clinical evaluation of the efficacy of photodynamic therapy in the treatment of erosive oral lichen planus: A case series.

BACKGROUND: Erosive oral lichen planus (EOLP) poses a substantial risk of malignant transformation into squamous cell cancer. The absence of established treatment gives way to alternative therapeutic strategies, including photodynamic therapy. The aim of the study was to evaluate the efficacy of PDT in the treatment of EOLP. METHODS: Twelve female patients aged 63-80 with 22 OLP lesions (16 on the buccal mucosa, 6 on gingiva and tongue), underwent authors' own PDT scheme with the use of 5% solution of 5-aminolevulinic acid (ALA) as photosensitizer. An ALA-saturated occlusive dressing was applied directly onto a lesion and surrounding mucosa 2h prior to illumination with a custom-made diode lamp (light of 630nm, dose of 300mW). After a series of 10 weekly illumination sessions the patients were monitored for 12 months. RESULTS: The mean size of lesions before treatment was 1.46cm2±1.44. The lesions on the buccal mucosa were smaller (1.06cm2±0.98) than those on the gingiva and tongue (2.63cm2±1.93). Post-treatment improvement encompassed 16 lesions, 5 of which were in remission. The mean reduction in size after 10-session therapy was 8,05%. The healing continued and further reduction in size (by 69.13%) took place during the 12-month observation: 39.62% of lesions within the buccal mucosa and full remission of all lesions on the gingiva and tongue. CONCLUSIONS: The results suggest that PDT offers non-invasive treatment of lesions in oral mucosa and may become an alternative and complementary method to those currently in use. Further studies involving larger groups of patients should be undertaken before it becomes routine practice.

[The importance of stem cells in the initiation and development of cancer]. / Znaczenie komórek macierzystych w inicjacji i rozwoju nowotworów.

Initiation of cancer may be the result of mutations occurring in stem cells, which causes blocking the differentiation of these cells. Many common properties of the stem cells and some tumor cells suggests that cancer stem cells may be responsible for the initiation and progression of cancer. The special properties of CSC is the ability to self-renewal and cell proliferation, which are the major cause of cancer recurrence and metastasis. Signaling pathways (Wnt, Notch, Shh) and pluripotency- connected transcription factors (Oct-4, Nanog) are primarily responsible for cell proliferation. Understanding the causes of initiation and progression of cancer is crucial for improving treatment of these life-threatening diseases.

[Photodynamic method of cancer diagnosis and therapy--mechanisms and applications]. / Metoda fotodynamiczna diagnostyki i leczenia nowotworów--mechanizmy i zastosowania.

Cancers are diagnosed in over few millions patients annually. Due to inter alia weak antigenicity as well as drug-fastness of most cancers, PDT can become an effective alternative or complementary method to surgery. Photodynamic method requires tree elements: photosensitizer, light energy source and oxygen. Photodynamic therapy is successfully used method in many clinics all over the world, also in Poland. PDT is used in treatment of oncological (cancers of head, neck, lung, oesophagus, pancreas, urinary bladder and ginecological cancers) as non oncological diseases (lischen sclerosus, staphylococcus aureus, papillomatosis).

Cytotoxicity of PP(Arg)2- and PP(Ala)2(Arg)2-based photodynamic therapy and early stage of apoptosis induction in human breast cancers in vitro.

Mitochondria are cell energetic centers where ATP is produced. They also play a very important role in the PDT as intracellular sites of photosensitizer localization. Photosensitizers gathering in mitochondria (like porphyrin derivatives used in this work) are more effective in tumor cell destruction. Moreover, it was assumed that di-amino acid substituents attached to porphyrin ring will strengthen the effectivity of interaction with membrane receptors of examined cells. MTT assay was performed to investigate the influence of PP(Arg)(2) and PP(Ala)(2)(Arg)(2)-based PDT on breast cancer cell viability for 24 h, 48 h and 120 h after cell irradiation. Then the influence of PP(Ala)(2)(Arg)(2)- and PP(Arg)(2)-mediated PDT on early mitochondrial apoptosis induction via measurements of the transmembrane mitochondrial potential changes was examined. Results showed that lower energy doses and maximal nontoxic photosensitizer doses of PP(Ala)(2)(Arg)(2) and PP(Arg)(2) applied in PDT can imply apoptotic cell death. It was confirmed that modification of the protoporphyrin IX by attaching two alanine substituents raised the efficiency of photodynamic therapy.

Cytotoxicity of PP(Arg)(2)- and Hp(Arg)(2)-mediated photodynamic therapy and early stage of apoptosis induction in prostate carcinoma in vitro.

Porphyrin photosensitizers tend to localize in mitochondria. The depolarization of mitochondrial membrane is one of the early stages of apoptosis and Laser Scanning Fluorescence Microscopy allows to determine changes in transmembrane mitochondrial potential under influence of PDT depending on the kind of photosensitizer (PP(Arg)(2), Hp(Arg)(2)), the energy dose (5, 10, 30 and 50 J/cm(2)) and time periods (24 and 48 hours after irradiation) in the LNCaP (lymphonodal metastasis of prostate carcinoma, the androgen dependent cell line). Cyototoxicity induced by PP(Arg)(2)- and Hp(Arg)(2)-based PDT depending on energy dose and time after irradiation in prostate carcinoma is determined with MTT. Generally, it was shown that lower energy doses induce greater changes in transmembrane mitochondrial potential. Hp(Arg)(2)-based PDT was more effective causing greater mitochondrial membrane depolarization and cell viability decrease in comparison to PP(Arg)(2)-mediated PDT (in the case of maximal nontoxic photosensitizer doses used).

Inhibition of cell growth induced by photosensitizer PP(Arg)2-mediated photodynamic therapy in human breast and prostate cell lines. Part I.

Photodynamic therapy can become an effective alternative method to surgery. The experiments reveal that using low photosensitizer doses and relatively low energy doses allow us to obtain effective results after PDT (to limit formation of colonies by investigated cancer cells). The prostate and breast cancer cell lines were investigated: MCF-7, a human breast cancer responsive to androgen therapy; MDA-MB231, a more aggressive human breast cancer non-responsive to androgen therapy; LNCaP, a lymphonodal metastasis of prostate carcinoma responsive to androgen therapy; DU-145, a human prostate cancer non-responsive to androgen therapy. Clonogenic assay shows that certain PP(Arg)(2) and light energy low doses stimulate the researched colony-forming cancer cells growth. Some low energy doses used during PP(Arg)(2)-mediated PDT also cause the increase in the colony-forming tumor cells. Among investigated cancer lines, MCF-7 exhibited the biggest sensibility towards PP(Arg)(2) and LNCaP the smallest one. PP(Arg)(2) based PDT is an effective method in colony growth limitation of breast cancer cell lines: MCF-7, MDA-MB231 and prostate cancer cell lines: LNCaP, DU-145.

Influence of photodynamic effect on biological activity of PBR-PP complexes.

The aim of this study was to examine the influence of photodynamic effect on biological activity of PBR-PP complexes. These measurements were performed in pH dependent environment. Constant concentration of solubilized receptor was titrated with increasing concentration of porphyrins (PPIX, Hp, PP(Arg)(2), Hp(Arg)(2), PP(Gly)(2), PP(Ala)(2), PP(Ser)(2), PP(Phe)(2)) and binding constants were calculated. PBP-PP mixtures were illuminated with 3 J, 5 J or 10 J of blue light and changes in protein fluorescence was recorded. Experimental data were fitted to weak and strong binding models. As a result for all derivatives weak binding model was the best fitted. The strongest binding showed PPIX in pH 7.4 and with pH drop binding constants showed greater values for all examined derivatives. Out of amino acid derivatives the strongest binding was noticed for PP(Gly)(2) and PP(Phe)(2) and for the last one pH influence was not observed.

Photodynamic therapy with 5-aminolevulinic acid and diamino acid derivatives of protoporphyrin IX reduces papillomas in mice without eliminating transformation into squamous cell carcinoma of the skin.

Photodynamic therapy (PDT) is used to treat malignant and nonmalignant diseases. It is also used for cosmetological skin treatment. PDT is generally considered to have a low risk of carcinogenicity. However, instances of nonmalignant human tumors turning malignant have been linked to PDT. In this study, we used 5-aminolevulinic (ALA) acid and 3 water soluble photosensitizers-PP(Arg)(2), PP(Ser)(2)Arg(2), PP(Ala)(2)Arg(2), all diamino acid derivatives of protoporphyrin IX-to treat benign papillomas in FVB/N mice induced by 7,12-dimethylbenz(a)anthracene (DMBA)-12-O-tetradecanoyl-phorbol-13-acetate (TPA). Of these drugs, ALA and PP(Arg)(2) were found the most efficient. PDT reduced the number of papillomas, but with increasing effectiveness of the drugs, the risk of malignant transformation of the papillomas into squamous cell carcinomas increased. The underlying mechanisms are not clear and further investigations are needed.

Interactions of protoporphyrin IX and its derivatives with benzodiazepine receptor.

The aim of this study was to determine binding affinities of the Peripheral benzodiazepine receptor (PBR) with protoporphyrin IX, haematoporphyrin (Hp), and two arginized derivatives: Hp(Arg)(2) and PP(Arg)(2). The quenching of protein fluorescence was used to measure association constants during titration of PBR solution with PP derivatives in a function of concentration. The experimental data was fitted to theoretical curves, assuming strong and weak binding model with one binding site. As a result of this early study affinity constants were determined. The highest affinity toward PBR showed PPIX and Hp, Hp(Arg)(2), PP(Arg)(2) lower, respectively. Changes in pH of protein solution resulted in decrease of association constants in nearly all examined derivatives.

Diamino acid derivatives of PpIX as potential photosensitizers for photodynamic therapy of squamous cell carcinoma and prostate cancer: in vitro studies.

Photodynamic therapy (PDT) is an alternative treatment modality involving light activated drugs, called photosensitizers (PSs), to treat cancer and non-cancerous conditions. The search for new compounds which might become effective PSs is the major direction for PDT development. In the present work we have studied the dark toxicity, intracellular localization and photodynamic properties of four potential, water soluble, second generation PSs--PP(Arg)(2), PP(Ser)(2)Arg(2), PP(Ala)(2)Arg(2), PP(Phe)(2)Arg(2), all diamino acid derivatives of protoporphyrin IX. Human prostate cancer (DU-145) and squamous carcinoma (A431) cells were used as experimental model. Among investigated compounds PP(Ser)(2)Arg(2) exhibited the lowest dark toxicity and the highest PDT effectiveness towards both cell lines. Fluorescence microscopy revealed the time-dependent changes in intracellular localization of the PS which were related to the phototoxicity. The results show that PP(Ser)(2)Arg(2) may be a potential PS for PDT.

Interactions of peripheral-type benzodiazepine receptor with diamino acid derivatives of protoporphyrin IX.

The aim of this study was to examine and determine binding affinities of the peripheral benzodiazepine receptor (PBR) with diamino acid derivatives of protoporphyrin IX with general structure PP(AA)(2) in a function of environmental pH changes. Fluorescence intensity was used as a measure of protein association constant. Decrease in protein fluorescence, after titration with increasing concentration of porphyrins, gave evidence of interactions between PBR and examined ligands. Experimental data were fitted to theoretical curves, assuming different models of interactions. All examined ligands were best fitted to weak binding model. Increase in environmental acidity resulted in changes in association constants. For all examined derivatives association constants were at least twice higher in pH 5.5 as compared to pH 7.4 and 6.5.

Photodynamic effect of protoporphyrin diarginate (PPArg2) on methicillin-resistant Staphylococcus aureus and human dermal fibroblasts.

The worldwide rise in the antibiotic resistance of bacteria forces the development of alternative antimicrobial treatments. A potential approach is photodynamic inactivation (PDI). The aim of the present study was to determine the phototoxicity of protoporphyrin diarginate (PPArg(2)) against methicillin-resistant Staphylococcus aureus and human dermal fibroblasts. Different concentrations (0 to 20 microM) of PPArg(2) and light dose of 6 J cm(-2) were tested. Cell viability was evaluated using the methylthiazoletetrazolium (MTT) assay. Incubation with 10 microM followed by illumination yielded a 3.6 log(10)-unit reduction in the viable count for Staphylococcus aureus. At the same experimental conditions, only 22.5% of the fibroblasts were photoinactivated. Protoporphyrin diarginate at concentrations up to 20 microM demonstrated no toxicity towards S. aureus or fibroblasts when not irradiated. These results suggest that the protoporphyrin diarginate exerts a high bactericidal effect against methicillin-resistant S. aureus strain without harming eukaryotic cells.

Bactericidal effect of photodynamic inactivation against methicillin-resistant and methicillin-susceptible Staphylococcus aureus is strain-dependent.

Staphylococcus aureus is one of the most important etiological factors responsible for nosocomial infections. Some of them may be life-threatening, especially in the case of immuno-compromised patients, causing bacteremia, endocarditis, sepsis or toxic-shock syndrome. Their multiresistance to antibiotics produces many therapeutic problems, and for this reason the development of a method alternative to antibiotic therapy is needed. It seems that photodynamic inactivation (PDI) may be an effective and alternative therapeutic option against both methicillin resistant (MRSA) and methicillin sensitive (MSSA) S. aureus strains. The aim of this study was to analyze the bactericidal effect of the PDI against 40 clinical MRSA and 40 MSSA clinical strains that were isolated from patients hospitalized in the Provincial Hospital in Gdansk. The ATCC strain 25904 has been used as a reference. Photodynamic inactivation by means of protoporphyrin diarginate as a photosensitizer was examined. It was observed that the bactericidal effect of the PDI was strain-dependent and ranged from 0 to 3 log(10)-unit reduction in viable counts. The mechanism underlying such a phenomenon is still not understood. Nevertheless, the correlation between the biofilm production ability and different strains response to PDI has been observed.

Contents of bioelements and toxic metals in the Polish population determined by hair analysis. Part IV. Adults aged 40 to 60 years.

The aim of this study was to define referential values of 5 basic bioelements (Ca, Mg, Zn, Cu, Fe) and 2 toxic metals (Pb and Cd) in the hair of Polish adults aged 40 to 60 years, based on the research conducted from 1991 to 2006 on a group of 3349 formally healthy women and men. The results of our biochemical tests were subjected to statistical analysis and they can be treated as referential norms for the concentrations of analyzed bioelements in the hair of Polish population. Our analyses confirm significant differences in accumulations of certain elements (Ca, Mg, Zn) in women's and men's hair. The hair element analysis of the tested women population indicates the age 49-50 as the one in which a significant decrease of hair Ca and Zn begins. Marked decrease of Ca and Zn in men begins 2-3 years later than in women. A slight tendency to decrease hair Mg in women and men during the period observed of 40-60 years of age was noticed. Significant differences (alpha = 0.05) between hair Cu contents in women and men were not observed. The increase of variances of hair Fe concentrations in women, especially after 50, has been observed. Significantly higher hair Pb and Cd concentrations were shown in men. The analysis of correlation confirms the significance (p < 0.050000) of synergistic interactions between bioelements: Ca-Mg (r = +0.7635), Ca-Zn (r = +0.36), Ca-Cu (r = +0.124), Ca-Fe (r = +0.054), Mg-Zn (r = +0.35), Mg-Cu (r = +0.11), Mg-Fe (r = +0.0757), Zn-Cu (r = +0.11), Cu-Fe (r = +0.0765). Significance (p < 0.050000) of antagonistic interactions of bioelements with toxic metals: Ca-Pb (r = -0.0965), Zn-Pb (r = -0.2476), Mg-Pb (r = -0.13), Zn-Cd (r = -0.0889) was established. Significance (p < 0.050000) of synergistic effects with toxic metals: Pb-Cd (r = +0.3322), Cu-Pb (r = +0.09), Fe-Pb (r = +0.1413), Fe-Cd (r = +0.1161) was confirmed. Adults in whose hair bioelements and hair toxic metal concentrations have shown values outlying from reference and who have too much toxic elements and too little bioelements should undergo further diagnostic tests since these results could be a sign of disturbances, that could lead to various diseases.

Bactericidal effect of photodynamic therapy against methicillin-resistant Staphylococcus aureus strain with the use of various porphyrin photosensitizers.

Photodynamic therapy (PDT) is based on photosensitizers activated by light of appropriate wavelength. Their activation leads to generation of singlet oxygen and free radicals responsible for the cytotoxic effect. The aim of this project was to compare the bactericidal effect of PDT using different porphyrin photosensitizers against a methicillin-resistant Staphylococcus aureus strain. Exogenous sensitizers (protoporphyrin IX and newly synthesized derivative, protoporphyrin diarginate) induced a 3 log10-unit reduction in bacterial viable counts. With the use of endogenous, ALA-induced porphyrins, a 1.6 log10-unit reduction was obtained. The sensitizers tested executed their antibacterial activity with no essential change in the antibiotic resistance pattern of the studied strain.

Contents of bioelements and toxic metals in the Polish population determined by hair analysis. Part III. Adults aged 20 to 40 years.

The aim of the present study was to define referential values of 5 basic bioelements (Ca, Mg, Zn, Cu, Fe) and 2 toxic metals (Pb and Cd) in the hair of Polish adults aged 20 to 40 years, based on the research conducted from 1991 to 2006 on a group of 5733 formally healthy subjects of both sexes. The results of our biochemical tests were subjected to statistical analysis and they can be treated as referential norms for the concentrations of analyzed bioelements in the hair of the Polish population. Our analyses confirm significant differences in accumulations of certain elements in women's and men's hair. We established that women's hair Ca, Mg, Zn (p = 0.000000) and hair Cu (p = 0.0001) concentrations are higher than they are in men. A lower mean of hair Fe concentrations in women were confirmed by some tests. Men have significantly higher (p = 0.0000) concentrations of toxic metals (hair Pb and Cd), which keep increasing with age. Analyses of correlations confirm significance (for the assumed level of significance alpha = 0.05) of synergistic interactions between bioelements: Ca-Mg (r = +0.70), Ca-Zn (r = +0.39), Ca-Cu (r = +0.14), Mg-Zn (r = +0.31), Mg-Cu (r = +0.10), Zn-Cu (r = +0.11), Zn-Fe (r = +0.07). Significance (a < 0.05) of antagonistic interactions with toxic metals: Ca-Pb (r = -0.13), Zn-Pb (r = -0.15), Mg-Pb (r = -0.15), Zn-Cd (r =-0.02) was confirmed. Significance (alpha < 0.05) of synergistic effects with toxic metals: Pb-Cd (r = +0.35), Fe-Pb (r = +0.16), Fe-Cd (r = +0.08) was also confirmed. Adults in whose hair bioelements and hair toxic metals concentrations have shown values outlying from reference and who have too many toxic elements and too few bioelements should undergo further diagnostic tests since the results could be a sign of disturbances that could lead to various diseases.

Biological activity of 5-aminolevulinic acid and its methyl ester after storage under different conditions.

5-Aminolevulinic acid (ALA) is a natural precursor of protoporphyrin IX (PpIX) and heme in cells. Photodynamic therapy (PDT) utilizes a metabolic imbalance in cancer cells, leading to increased PpIX generation from exogenous ALA. Due to chemical instability of ALA in therapeutic concentrations at pH values larger than 5.0 and at high temperatures, it looses its activity by spontaneous dimerization to 2,5-dicarboxyethyl-3,6-dihydropyrazine (DHPY). ALA esters are now supplementing ALA in PDT, but little is known about their stability. We have studied the stability of ALA and its methyl ester (MAL) stored under different conditions (temperatures, pH values) by measuring their ability to generate PpIX. 100mM solutions of both compounds were found to be stable at pH 4 and at 4 degrees C. However, at pH 5.5 they lost almost 10% of the initial activity during 5days of storage at 4 degrees C. The fastest decay of ALA and MAL was seen at pH 7.4 and at 37 degrees C, and followed first order kinetics. At pH 7.4 and at 4 degrees C MAL lost its PpIX producing ability more slowly than at 37 degrees C. Our work shows that solutions should be prepared immediately before use and stored at low temperatures. The pH of stock solutions should not exceed 5.