Concentrations of metalloproteinase-1 in patients with morphea treated with phototherapy: a preliminary study.

Introduction: Morphea (localized scleroderma) is a rare, chronic, inflammatory connective tissue disease, characterized by immune system dysfunction, vasculopathy and skin fibrosis. One of the most effective treatments is phototherapy. Phototherapy has been found to be effective in treating localized scleroderma by inducing the expression of metalloproteinase-1. Aim: To compare the concentrations of metalloproteinase (MMP-1) before psoralen and ultraviolet A (PUVA) and ultraviolet A1 (UVA1) treatments in the serum of patients with morphea. Material and methods: The observational study was conducted in one research centre and included patients with generalised morphea who were treated with PUVA and UVA1 phototherapies. The mean age of all morphea patients included in the study was 55.7 years. The levels of MMP-1 were examined by ELISA (The Biorbyt Human MMP-1 ELISA - Enzyme-Linked Immunosorbent Assay). Results: The study showed that patients treated with PUVA and UVA1 had an improvement based on clinical measures, resulting in a reduction of clinical score. However, we did not observe statistically significant differences in MMP-1 concentrations before and after treatment. Limitations: The study sample was relatively small. Further studies on a larger group of patients would be beneficial. Conclusions: Our data suggest that there is a possible correlation between MMP-1 concentrations and phototherapy. MMP-1 levels were found to be increased following phototherapy treatment, which may suggest a correlation with better response to treatment in patients with morphea. However, further research is needed.

Effectiveness of PUVA vs. UVA1 phototherapy in the treatment of morphea patients.

Introduction: Morphea (localized scleroderma) is an inflammatory connective tissue disease, characterized by immune system dysfunction, vasculopathy and skin fibrosing. Phototherapy has been found to be effective in treating localized scleroderma. Psoralen + ultraviolet A (PUVA) and ultraviolet A1 (UVA1) phototherapy significantly enriched therapeutic possibilities. Aim: To compare the clinical effect of PUVA photochemotherapy and UVA1 phototherapy and to evaluate the treatment response rates. Material and methods: It was a retrospective one-centre research and observational study of all morphea patients treated with PUVA and UVA phototherapy. We reviewed phototherapy notes along with electronic and paper case records for all patients with morphea treated with PUVA and UVA1 phototherapy from January 2010 to December 2019. Results: The study shows that patients in both groups experienced improvement based on clinical measures, resulting in a reduction in the clinical score in all groups. There is positive short- and long-term efficacy of UVA1 and PUVA phototherapy in patients with morphea. There were no statistical differences between the treatment response rates. Limitations: We had a relatively small study sample and it was a retrospective, observational study. Conclusions: Our data suggest that ultraviolet PUVA and UVA1 should be considered for the treatment of morphea with disseminated lesions or not responding to topical treatment. UVA1 is free of side effects linked to oral psoralens such as nausea, vomiting, photokeratosis, but we showed that there was no statistical advantage in the effectiveness of both. UVA1 phototherapy is, however, a less accessible form of treatment, available in the centres of higher quality.

Secretory Products in Petals of Centaurea cyanus L. Flowers: A Histochemistry, Ultrastructure, and Phytochemical Study of Volatile Compounds.

(1) Background: Centaurea cyanus L. is a medicinal plant whose flowers are widely used in herbal medicine. The aim of the study was to localise flower tissues that are responsible for the production of secretory products in petals and to analyse the volatile compounds. The volatile compounds of the flowers of this species have not been investigated to date. (2) Methods: Light, fluorescence, scanning and transmission electron microscopy techniques were used in the study. Lipophilic compounds were localised in the tissues using histochemical assays. Volatile compounds were determined with the use of solid phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). (3) Results: The study showed production of secretion in the petal parenchyma, whose ultrastructure has features of a secretory tissue. The lipophilic secretion was localised in the cells and intercellular spaces of the parenchyma and in the walls and surface of epidermal cells, where it accumulated after release through cuticle microchannels. Sesquiterpenes were found to constitute the main group of volatile compounds, with the highest content of ß-caryophyllene (26.17%) and α-humulene (9.77%). (4) Conclusions: Given the presence of some volatile components that are often found in resins (caryophyllene, delta-cadinene) and the abundant secretion residues on the epidermal surface, we suppose that the C. cyanus secretion released by the flowers is a resinaceous mixture (oleoresin), which is frequently found in plants, as shown by literature data. This secretion may play an important role in the therapeutic effects of C. cyanus flowers.

Phenolic Constituents of Lamium album L. subsp. album Flowers: Anatomical, Histochemical, and Phytochemical Study.

Flos Lamii albi has a high biological activity and is widely used in herbal medicine. The aim of the study was to characterize the secretory structures present in Lamium album subsp. album corolla and the location of phenolic compounds. Additionally, we carried out qualitative phytochemical analyses of flavonoids and phenolic acids. Light, fluorescence, and scanning electron microscopy were used to analyze the structure of the floral organs. The main classes of phenolic compounds and their localization were determined histochemically. Phytochemical analyses were performed with high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC). Six types of glandular trichomes were found which contained flavonoids, phenolic acids, and tannins. The phytochemical studies demonstrated the presence of caffeic, chlorogenic, ferulic, gallic, p-coumaric, protocatechuic, syringic, gentisic, and vanillic phenolic acids as well as rutoside, isoquercetin, and quercetin flavonoids. The corolla in L. album subsp. album has antioxidant properties due to the presence of various polyphenols, as shown by the histo- and phytochemical analyses. The distribution and morphology of trichomes and the content of phenolic compounds in the corolla have taxonomic, pharmacognostic, and practical importance, facilitating the identification of the raw material.

Influence of silver content on rifampicin adsorptivity for magnetite/Ag/rifampicin nanoparticles.

Magnetite nanoparticles (NPs) decorated with silver (magnetite/Ag) are intensively investigated due to their application in the biomedical field. We demonstrate that the increase of silver content on the surface of nanoparticles improves the adsorptivity of antibiotic rifampicin as well as antibacterial properties. The use of ginger extract allowed to improve the silver nucleation on the magnetite surface that resulted in an increase of silver content. Physicochemical and functional characterization of magnetite/Ag NPs was performed. Our results show that 5%-10% of silver content in magnetite/Ag NPs is already sufficient for antimicrobial properties against Streptococcus salivarius and Staphylococcus aureus. The rifampicin molecules on the magnetite/Ag NPs surface made the spectrum of antimicrobial activity wider. Cytotoxicity evaluation of the magnetite/Ag/rifampicin NPs showed no harmful action towards normal human fibroblasts, whereas the effect on human embryonic kidney cell viability was time and dose dependent.

The New TLC Method for Separation and Determination of Multicomponent Mixtures of Plant Extracts.

The new mode of two-dimensional gradient thin layer chromatography (MGD-2D TLC) has been presented. Short distance development of sample in the first dimension leads to formation of the preconcentrated narrow zones. They are consecutively separated in the second dimension with the mobile phase gradient in several steps of development until the eluent reaches the further end of the chromatographic plate. The use of the above-mentioned technique allows isolating and then identifying the compounds of various polarity from the multicomponent mixture. The practical application of two-dimensional gradient thin layer chromatography has been performed for isolation of the two plant (Juniperus and Thymus) oils components as the examples of test mixtures. The experiments have been carried out with the use of silica gel plates as well as a normal phase condition. The results of solute separation with isocratic one-dimensional thin layer chromatography system have been compared with those of two-dimensional gradient system. It has been observed that application of the latter mode leads to almost triplicated number of zones in comparison with the former one. It is purposeful to apply the proposed mode to control the purity of the dominant component or components of the mixture.

Herbal preparation extract for skin after radiotherapy treatment. Part One--Preclinical tests.

Naran R is a herbal composition made of Plantago lanceolate folium, Malvae arboreae flos, Calendulae flos, Chamomillae inflorescentia, Lamii albi flos to prepare compresses or to wash skin with inflammations. The extract of this preparation is mixed to be applied as an ointment on patients' skin after radiotherapy. Experiments performed in vitro are part of pre-clinical tests with Naran R ointment. This study examined the impact of the plant composition for ethanol-water extract on human skin fibroblasts (HSF) culture. Samples of extract, prepared from patented amounts of herbs, were in the range of 25-225 µg/mL. Six methods were applied: standard spectrophotometric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, neutral red (NR) uptake assay, DPPH free radical scavenging test, labeling of cytoskeleton F-actin, staining of argyrophilic nucleolar organizer regions (AgNORs) and trypan blue coloration. The extract concentration 75 µg/mL was established as safe for application on human skin. In labeling of F-actin with rhodamine-phalloidin dye at this concentration the cytoskeleton was stable. The extract did not influence the membrane stability and had positive influence on the proliferation activity. It was confirmed in AgNOR test during incubation with extract, which led to formation of larger amount of smaller nucleolins. In DPPH scavenging activity test, the extract revealed over 8% higher free-radical scavenging activity in comparison to control. After trypan blue staining, the extract in concentration 125 µg/mL significantly lowered the cell viability. When the cytotoxic and anti-proliferative activity of the extracts were analyzed, MTT and Neutral Red (NR) methods were used. The cells' viability was maintained on a constant level (80-110%) after 24, 48 and 72 h of incubation. During all time of NR test (72 h) and even when 225 µg/mL of extract was applied, the viability of cells was in range 80-110% of control. Positive influence of the extract on investigated cells structure and proliferation, lack of toxicity and increasing anti-oxidant activity enable to consider this preparation as a natural remedy with potential application in skin therapy after radiation.

Assessment of eyebright (euphrasia officinalis L.) extract activity in relation to human corneal cells using in vitro tests.

BACKGROUND: Euphrasia officinalis L. is an herb traditionally used in folk medicine, mainly in the treatment of eye disorders. AIMS: The present study analyzed the activity of three extracts of E. officinalis L. (ethanol, ethyl acetate and heptane) on cultured human corneal epithelial cells (10.014 pRSV-T). STUDY DESIGN: In vitro study. METHODS: Toxicity, free radical scavenging activity and the immunomodulatory effects of the extracts were tested using the thiazolyl blue tetrazolium bromide (MTT) or Neutral Red, 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and ELISA tests, respectively. Moreover, nitric oxide levels and cytoskeleton architecture were analyzed after corneal cell incubation with the plant extracts. RESULTS: We show that the biological effect depended on both the concentration and the extraction solvent used. Heptane extracts, distinct from those in ethanol and ethyl acetate, were toxic to 10.014 pRSV-T cells at low concentrations (25 µg/mL) and did not demonstrate free radical scavenging effects. All tested extracts decreased pro-inflammatory cytokine expression (IL-1ß, IL-6 and TNF-α) and also anti-inflammatory IL-10 expression by human corneal cells when the extracts were added to the cell culture medium for 24 h. CONCLUSION: In conclusion, we show that the promising effects of the application of E. officinalis L. preparations as a supplementary therapy for eye disorders are associated with the ethanol and ethyl acetate extracts, not the heptane extract.

[Valproic acid concentration in the blood serum of pregnant women and umbilical cord blood in relation to the condition of the newborn--case studies]. / Stezenie kwasu walproinowego w surowicy krwi kobiet ciezarnych oraz krwi pepowinowej w odniesieniu do stanu noworodka--analiza przypadków.

Epilepsy is one of the most common neurological diseases in the world. One of the most difficult clinical problems associated with the disease is to treat pregnant women because the use of antiepileptic drugs increase the risk of birth defects in the fetus. The second most common use in pregnant women is an antiepileptic drug valproic acid. Its use is associated with an increased risk of serious birth defects such as neural tube defects, heart defects, cleft palate, urinary tract defects, limb defects, specific syndromes that cause dysmorfizm face or abnormalities of the reproductive organs and developmental disorders affecting cognitive and behavioral functions. To minimize the risk to the fetus can be through the use of contraception and planning for pregnancy, taking valproic acid monotherapy, at a dose of less than 1000 mg/day in 2-3 divided doses, folic acid supplementation, close monitoring during pregnancy and full cooperation with doctors: a gynecologist and neurologist. The aim of this study was to measure the concentrations of valproic acid in maternal serum and cord serum and a reference to the results of the newborn. The concentrations of valproic acid in the serum was performed using high performance liquid chromatography method with fluorometric detection. Valproic acid concentration in the blood serum of patients housed or slightly exceed the therapeutic concentration. In all samples of umbilical cord blood serum were detected, no drug found to have birth defects. Born children should remain under control in order to detect possible birth defects that cannot be detected immediately after birth.

Aloe vera extract activity on human corneal cells.

CONTEXT: Ocular diseases are currently an important problem in modern societies. Patients suffer from various ophthalmologic ailments namely, conjunctivitis, dry eye, dacryocystitis or degenerative diseases. Therefore, there is a need to introduce new treatment methods, including medicinal plants usage. Aloe vera [Aloe barbadensis Miller (Liliaceae)] possesses wound-healing properties and shows immunomodulatory, anti-inflammatory or antioxidant activities. MATERIALS AND METHODS: NR uptake, MTT, DPPH• reduction, Griess reaction, ELISA and rhodamine-phalloidin staining were used to test toxicity, antiproliferative activity, reactive oxygen species (ROS) reduction, nitric oxide (NO) and cytokine level, and distribution of F-actin in cells, respectively. AIM: The present study analyzes the effect of Aloe vera extracts obtained with different solvents on in vitro culture of human 10.014 pRSV-T corneal cells. RESULTS: We found no toxicity of ethanol, ethyl acetate and heptane extracts of Aloe vera on human corneal cells. No ROS reducing activity by heptane extract and trace action by ethanol (only at high concentration 125 µg/ml) extract of Aloe vera was observed. Only ethyl acetate extract expressed distinct free radical scavenging effect. Plant extracts decreased NO production by human corneal cells as compared to untreated controls. The cytokine (IL-1ß, IL-6, TNF-α and IL-10) production decreased after the addition of Aloe vera extracts to the culture media. DISCUSSION AND CONCLUSIONS: Aloe vera contains multiple pharmacologically active substances which are capable of modulating cellular phenotypes and functions. Aloe vera ethanol and ethyl acetate extracts may be used in eye drops to treat inflammations and other ailments of external parts of the eye such as the cornea.

[Development and validation of method for the determination of cynarin, luteolin in plasma]. / Opracowanie i walidacja metody oznaczania cynaryny, luteoliny w osoczu.

The aim of this study was to develop and validate the method of cynarin and luteolin, the main constituents of artichoke (Cynara scolymus L.) leaf extract, determination in plasma. The compounds were separated using the high-performance liquid chromatography technique with diode array detection (HPLC-DAD). The analysis was preceded by liquid-liquid extraction using as the extracting agent ethyl acetate. The HPLC separation was performed on C18 column under gradient conditions using a mobile phase - 0,05% trifluoroacetic acid in water and methanol. The detector was set at lambda=330 nm. The validation was related to linearity, sensitivity (LOD and LOQ), accuracy and repeatability. In the validated method the linearity was achieved within concentration range 1,5625 - 50,0 microg/cm3 for the cynarin (R2=0,9989) and 1,5625 - 200,0 microg/cm3 for the luteolin (R2=0998). The limits of detection for cynarin and luteolin was: 0,75 microg/cm3 and 0,1 microg/cm3 and the limits of quatification: 2,25 microg/cm3 and 0,2 microg/cm3, respectively. Coefficient of variation for the inter-day and the intra-day analysis, which is a precision and accuracy parameter, do not exceed 10%. Recovery was 67% for the cynarin and 96% for the luteolin. The practical application of this method was proved by analysis of plasma samples from rats. The animals were administrated artichoke leaf extract - orally and intraperitoneally at a dose of 3 g/kg body weight or pure substances - intraperitoneally at a dose 1 mg/kg of luteolin and 0,5 mg/kg of cynarin. The presence of investigated compounds was proved only in samples after intraperitoneal administration of pure substances. The developed method is used to determine simultaneously cynarin and luteolin, after intraperitoneal administration of pure compounds.

[Artichoke--herbal drug]. / Karczoch zwyczajny--lek roslinny.

The liver is the gland most vulnerable to the toxic effects of xenobiotics, as responsible for their metabolism. Significant impact on the functioning of this gland has a style of life: alcohol consumption, diet with high fats ingredients and prooxidative substances and synthetic drugs. Very improtant aspect in herbal medicaments is protective properties on parenchymal organ-damaging. Concomitant intake of plant extracts containing cytoprotective compounds, may increase the efficacy of treatment and reduce side effects. One of the plants of the hepatoprotective action is artichoke (Cynara scolymus L.). Artichoke with multiple therapeutic properties and practically no side effects is recommended not only in disorders of the liver, but also in the prevention of atherosclerosis and hyperlipidemia or dyspeptic disorders.

[Artichoke--untapped potential of herbal medicine in the treatment of atherosclerosis and liver diseases]. / Karczoch zwyczajny- niewykorzystane mozliwosci leku roslinnego w terapii miazdzycy i chorób watroby.

Substances of natural origin are the subject of growing interest on the part of both researchers and doctors. One of the well known herbal medicines extensively examined in terms of clinical and pharmacological is artichoke (Cynara scolymus L.), which was used in European medicine from the 18th century. His multidirectional treatment is a documented fact and it is associated with treatment of dyspepsia, influence of active substances contained in artichoke on plasma lipid levels and with a strong antioxidant effect of the artichoke extract--due to this properties, artichoke compounds have a protective effect on liver cells.

The effect of ursolic and oleanolic acids on human skin fibroblast cells.

In this article, we look at how ursolic and oleanolic acids can be used for the purpose of quality control of natural products used in dermatocosmetology as well as of various other therapeutic preparations. Ursolic acid (UA) and oleanolic acid (OA) are pentacyclic triterpenes and they are constituents of many medicinal herbs. In this study, we analyzed the cytotoxic and anti-proliferative activity of OA and UA against normal human skin fibroblasts (HSF). Additionally, the scavenging activity of free radicals of both acids was analyzed. The sensitivity of cells to OA and UA activity was determined using a standard spectrophotometric (MTT) assay. The free radical scavenging activity of OA and UA was measured using the DPPH• test. The F-actin cytoskeletal proteins organization was analyzed using TRITC-phalloidine fluorescent staining. The cytotoxic activity of the analyzed acids was determined using Neutral Red (NR) uptake assay. Of the two isomeric compounds, UA showed a higher cytotoxic activity against HSF cells than did OA. Our investigations showed that OA, in view of its non-toxic nature, may be used as a supplementary factor for dermal preparations.