Györgyi Horváth

PACAP is an endogenous protective factor-insights from PACAP-deficient mice.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a widespread neuropeptide with a diverse array of biological functions. Not surprisingly, the lack of endogenous PACAP therefore results in a variety of abnormalities. One of the important effects of PACAP is its neuroprotective and general cytoprotective role. PACAP protects neurons and other tissues against ischemic, toxic, and traumatic lesions. Data obtained from PACAP-deficient mice provide evidence that endogenous PACAP also has protective functions. Mice lacking PACAP are more vulnerable to different in vitro and in vivo insults. The present review summarizes data on the increased sensitivity of PACAP-deficient mice against harmful stimuli. Mice lacking PACAP respond with a higher degree of injury in cerebral ischemia, autoimmune encephalomyelitis, and axonal lesion. Retinal ischemic and excitotoxic injuries also produce increased cell loss in PACAP-deficient mice. In peripheral organs, kidney cell cultures from PACAP-deficient mice are more sensitive to oxidative stress and in vitro hypoxia. In vivo, PACAP-deficient mice have a negative histological outcome and altered cytokine response in kidney and small intestine ischemia/reperfusion injury. Large intestinal inflammation, toxic lesion of the pancreas, and doxorubicin-induced cardiomyopathy are also more severe with a lack of endogenous PACAP. Finally, an increased inflammatory response has been described in subacute endotoxin-induced airway inflammation and in an oxazolone-induced allergic contact dermatitis model. In summary, lack of endogenous PACAP leads to higher vulnerability in a number of injuries in the nervous system and peripheral organs, supporting the hypothesis that PACAP is part of the endogenous cytoprotective machinery.

Determination of polyphenolic compounds by liquid chromatography-mass spectrometry in Thymus species

Polyphenolic compounds represent a wide group of phytochemicals, including well-known subgroups of phenolic acids, flavonoids, natural dyes, lignans etc., which are produced by plants. These natural bioactive compounds possess a variety of beneficial effects including antioxidant and anticarcinogenic activities, protection against coronary diseases as well as antimicrobial properties. Thymus species have already been reported as sources of different phenolic acids and flavonoids. Moreover, the composition and content of flavonoids in Thymus species play important role as taxonomic markers providing distinction of species. High-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) and on-line mass spectrometry (ESI-MS) method was used for analysis. The method was evaluated for a number of validation characteristics (repeatability and intermediate precision, LOD, LOQ, calibration range, and recovery). The polyphenolic pattern of five native Hungarian Thymus species (T. glabrescens Willd., T. pannonicus All., T. praecox Opiz, T. pulegioides L., and T. serpyllum L.) was characterized. The dominant compound was rosmarinic acid, which ranged between 83.49 μg g(-1) and 1.436 mg g(-1). Other phenolic acids (ferulic acid, caffeic acid and its other derivatives, chlorogenic acid and p-coumaric acids) were present in every examined Thymus species, as well as flavanones: naringenin, eriodictyol and dihydroquercetin; flavones: apigenin and apigenin-7-glucoside, flavonols: quercetin and rutin. The polyphenolic pattern was found to be a useful additional chemotaxonomic tool for classification purposes and determination of the locality of origin.

Agonistic Behavior of PACAP6-38 on Sensory Nerve Terminals and Cytotrophoblast Cells

The effects of pituitary adenylate cyclase activating polypeptide (PACAP) are mediated through G-protein-coupled receptors, the specific PAC1 receptor and VPAC1 and VPAC2 receptors which bind vasoactive intestinal peptide with similar affinity. Based on binding affinity studies, PACAP6-38 was discovered as a potent antagonist of PAC1 and it has been used by hundreds of studies as a PACAP antagonist. Recently, we have found that in certain cells/tissues, PACAP6-38 does not antagonize PACAP-induced effects, but surprisingly, it exerts similar actions to PACAP1-38, behaving as an agonist. In the present study, we report on the agonistic behavior of PACAP6-38 on neuropeptide release from sensory nerves of the isolated rat trachea and on the MAPK signaling pathways in cytotrophoblast cells. In isolated rat tracheae, PACAP6-38, similarly to PACAP1-38, induced significant inhibitory effects on the release of three simultaneously measured sensory neuropeptides, substance P, calcitonin gene-related peptide, and somatostatin evoked by both chemical excitation and electrical field stimulation of capsaicin-sensitive afferents. Effects of PACAP6-38 were the same as those of PACAP1-38 on MAPK signaling in human cytotrophoblast cells. Western blot analysis showed that both peptide forms stimulated ERK1/2 and JNK phosphorylation, while they both inhibited p38 MAPK phosphorylation. The most pronounced effects were observed when both peptides were present. In summary, our results show that PACAP6-38, which is a PACAP receptor antagonist in most cells/tissues, can behave as an agonist in other systems. The increasing interest in the effects of PACAP requires further studies on the pharmacological properties of the peptide and its analogues.

Chloroparva pannonica gen. et sp. nov. (Trebouxiophyceae, Chlorophyta) - a new picoplanktonic green alga from a turbid, shallow soda pan

Somogyi B., Felföldi T., Solymosi K., Makk J., Homonnay Z.G., Horváth G., Turcsi E., Böddi B., Márialigeti K. and Vörös L. 2011. Chloroparva pannonica gen. et sp. nov. (Trebouxiophyceae, Chlorophyta) – a new picoplanktonic green alga from a turbid, shallow soda pan. Phycologia 50: 1–10. DOI: 10.2216/10-08.1 We describe Chloroparva pannonica Somogyi, Felföldi & Vörös gen. et sp. nov., a new trebouxiophycean picoplanktonic alga isolated from a turbid, shallow soda pan in Hungary. The cells are spherical to oval, less than 2 µm in diameter, with simple ultrastructure typical to small green algae. Cells divide by autosporulation, forming two daughter cells per autosporangium. Cell wall structure consists of an outer trilaminar layer, an inner microfibrillar layer and an electron-transparent layer covering the plasma membrane. The trilaminar layer of the mother cell wall often persists around the autospores. Typical chlorophyte pigments have been found, including chlorophyll a and b and lutein as the dominant carotenoid. The main fatty acid was oleic acid. The phylogenetic position of the new chlorophyte confirms the proposal of a new genus within the Trebouxiophyceae. Based on its 18S rRNA gene sequence, this isolate is distantly related to Nannochloris eucaryotum UTEX 2502, Chlorella minutissima C-1.1.9 and C. minutissima SAG 1.80 (≤ 97.6% 18S rRNA gene pairwise similarities).

Essential Oil Composition of Three Cultivated Thymus Chemotypes from Hungary

Abstract The essential oils of three Hungarian cultivated thymes (Thymus vulgaris L., Thymus x citriodorus (Pers.) Schreb, Thymus x citriodorus “archers gold”) were obtained by steam distillation of air-dried plant materials and analyzed by GC and GC/MS. Three different chemotypes were found. In the oil of T. vulgaris thymol (45.6%), in T. x citriodorus geraniol (39.2%) and in T. x citriodorus, “archers gold,” carvacrol (43.5%) was the main component. Among the other constituents, p-cymene, β-caryophyllene, geranial, 1,8-cineole and γ-terpinene were characterized.

Changes in the expression of pituitary adenylate cyclase-activating polypeptide in the human placenta during pregnancy and its effects on the survival of JAR choriocarcinoma cells.

Pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide with survival-promoting actions, has been observed in endocrine organs and is thought to play a role in reproductive functions, including pregnancy. PACAP occurs in two forms, 27 and 38 amino acid residues, with PACAP38 being the predominant form in human tissues. In the present study, we determined the concentrations of PACAP38 and PACAP27 in first-trimester and full-term human placentas using radioimmunoassay. We found high levels of PACAP38 and lower levels of PACAP27 in different parts of the full-term human placenta. PACAP38 content increased in the placenta during pregnancy, both on the maternal side and on the fetal side. The effects of PACAP on the survival of JAR human choriocarcinoma cells were investigated using flow cytometry and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) cell viability assay in cells exposed to the widely used chemotherapeutic agent methotrexate (MTX). It was found that PACAP neither influenced the survival of JAR cytotrophoblast cells nor affected cellular response to the death-inducing effect of the chemotherapeutic agent MTX. The present observations further support the significance of PACAP in the human placenta. The observation that PACAP did not influence the effects of MTX may have future clinical importance, showing that PACAP does not decrease the effects of certain chemotherapeutic agents.

Characterization and TLC-bioautographic detection of essential oils from some Thymus taxa. Determination of the activity of the oils and their components against plant pathogenic bacteria

The chemical composition of the oils of four thyme (Lamiaceae) chemotypes (Thymus vulgaris L., Thymus serpyllum L., Thymus x citriodorus (Pers.) Schreb., and Thymus x citriodorus “Archer’s Gold”) has been determined by gas chromatography (GC). Thymol was the main component of the oils of Thymus vulgaris and Thymus serpyllum, geraniol was the main component of the oil of Thymus x citriodorus, and carvacrol was the main component of the oil of Thymus x citriodorus “Archer’s Gold”. The bioactivity of the volatile oil of Thymus vulgaris and of the three main components of the oils against Gram negative plant pathogenic bacteria was examined by direct bioautography. They had an inhibitory effect on all of the test microorganisms. Two bacterial strains (Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. phaseolicola) were most sensitive in the bioautographic system and use of these bacteria would considerably shorten the process of bioautographic detection. The two antibiotics gentamycin and streptomycin were used as controls.

Presence of Pituitary Adenylate Cyclase Activating Polypeptide and Its Type I Receptor in the Rat Kidney

Pituitary adenylate cyclase activating polypeptide (PACAP), a multifunctional neuropeptide, has 2 active forms, PACAP38 and PACAP27. It is now well-established that PACAP has several actions also in peripheral organs, including renoprotective effects. The peptide itself has not been previously identified in the rat kidney. The first aim of our study was to identify PACAP in the rat kidney using mass spectrometry and radioimmunoassay (RIA). Receptor mRNA and binding studies revealed the existence of all 3 PACAP receptors (PAC1, VPAC1, and VPAC2) in the kidney, but their exact localization in histologic sections was not evident. Because most of the cytoprotective effects of PACAP relate to its specific PAC1 receptor, our second aim was to identify the cell types wherein the PAC1 receptor is expressed in the rat kidney. Mass spectrometry revealed the presence of PACAP38 in the kidney. RIA measurements showed both PACAP38- and PACAP27-like immunoreactivities in kidney homogenates, with PACAP38 being dominant. Immunohistochemistry revealed PAC1 receptor-like immunoreactivity in kidney sections, mainly expressed in cortical tubular epithelial cells. These results showed PACAP to be endogenously present in the kidney. The tubular localization of the PAC1 receptor provides the basis for the renal effects of the peptide under physiologic and pathologic conditions.

Investigation of thyme (Thymus vulgaris L.) essential oil by use of the BioArena system

The composition of the essential oil of Thymus vulgaris L. has been determined by GC-FID and GC-MS. Because separation of thymol, carvacrol, and linalool, components of the essential oil, was more efficient by overpressured layer chromatography (OPLC) than by conventional thin-layer chromatography (TLC), the forced flow technique was used before biological detection. All three test compounds had antibacterial effect against the phytopathogenic bacterium Pseudomonas syringae pv. maculicola, in bioautography, although in essential oil thymol was present in sufficient quantity to produce an inhibiting zone in the adsorbent layer. In BioArena investigations, when reduced glutathione as a formaldehyde (HCHO) capturer was dissolved in the cell suspension before bioautographic exposure to the essential oil, the characteristic inhibiting activity of thymol and carvacrol against Bacillus subtilis soil bacteria was reduced, whereas the presence of the HCHO precursors NG-monomethyl-l-arginine or Nε-monomethyl-l-lysine enhanced their antibacterial effect. These results suggest that HCHO and its reaction products may be involved in the antibacterial activity of thymol and carvacrol.

Effects of Pituitary Adenylate Cyclase Activating Polypeptide on the Survival and Signal Transduction Pathways in Human Choriocarcinoma Cells

The pituitary adenylate cyclase activating polypeptide (PACAP) has several effects in endocrine and reproductive organs, including the placenta. PACAP is generally known as a survival‐promoting peptide acting on divergent signal transduction pathways. However, its effects on the survival and signaling mechanisms of trophoblast cells are not known. In the present study we found that 1‐h pretreatment with PACAP38 did not significantly influence the survival of JAR cytotrophoblast cells. However, the survival rate of cells exposed to oxidative stress or CoCl2‐induced in vitro hypoxia showed a significant further decrease in PACAP‐treated cells, implying that PACAP sensitizes the cells to these stressors. This was not observed in the case of lipopolysaccharide or ethanol treatment. Western blot data revealed that, in cells exposed to oxidative stress, PACAP treatment decreased phosphorylation of all extracellular signal‐regulated kinase (ERK), phospho‐jun N‐terminal kinase (JNK), protein kinase B, p38 mitogen‐activated protein kinase (MAPK), and phospho‐glycogen synthase kinase (GSK) and the expression of bax. The overall effect seems to be a sensitizing effect in almost all examined pathways when oxidative stress was applied, which may explain the enhancing effect of PACAP on cell death in contrast to most other cell types examined so far. Our data show that the signaling mechanism of PACAP may be different in trophoblast cells to that observed in other cell lines.