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Histaminski sustav je uključen u regulaciju različitih fizioloških funkcija, kao što su izlučivanje želučane kiseline, neurotransmisija i imunomodulacija. Histaminski sustav u užem smislu obuhvaća četiri podtipa histaminskih receptora i njihove izoforme putem kojih histamin, endogeni biogeni amin ostvaruje svoje biološke učinke. Cilj ovog diplomskog rada bio je opisati ulogu histaminskog sustava u zdravlju i bolesti kroz prikaz najznačajnijih znanstvenih rezultata koja su dovela do sinteze lijekova koji svoje terapijske učinke ostvaruju modulirajući funkciju tog sustava. Tijekom nekoliko desetljeća postignut je značajan napredak u razumijevanju uloge histaminskog sustava u različitim patofiziološkim procesima, od ulkusa i alergija, do bolesti središnjeg živčanog sustava. Lijekovi koji blokiraju histaminske H1- i H2-receptora već se dugi niz godina uspješno koriste u liječenju čestih medicinskih stanja, kao što su alergijske reakcije i ulkusna bolest. Dodatno je identifikacija histaminskih H3- i H4-receptora otvorila nova područja za istraživanje fiziološke uloge histaminskog sustava te razvoj novih generacija lijekova. Potencijalne nove skupine lijekova, H3- i H4-antagonisti predmet su brojnih pretkliničkih istraživanja, s ciljem utvrđivanja njihove djelotvornosti i sigurnosnog profila te optimiranja fizikalno-kemijskih i farmakoloških svojstava. Za neke od najperspektivnijh spojeva u kliničkim se studijama ispituje potencijalna terapijska vrijednost, i to u stanjima kao što su narkolepsija, kognitivni poremećaji, pretilost, shizofrenija, bol, ovisnost. The histamine system regulates a variety of physiological functions, such as secretion of hydrochloric acid, neurotransmission and immunomodulation. In its core, the histamine system includes four subtypes of histamine receptors and its isoforms by which histamine, an endogenous biogenic amine, carries out its biological effects. The aim of this master’s thesis was to describe the role of histamine system in health and illnesses while reviewing most prominent scientific results that led to development of medications which achieve their effects by modulating the function of the system. During the last few decades great progress has been made in understanding the role of histamine system in many pathophysiological processes, from ulcer and allergies, to diseases of central nervous system. For many years drugs that block histamine H1- and H2-receptors are successfully used in treatment of common medical conditions, such as allergic reactions and peptic ulcer disease. Additionally, the identification of H3- and H4–receptors opened new areas for research about physiological role of the histamine system and development of new drugs. The potential new generations of H3- and H4-antagonists drugs are the subject of many preclinical researches with aim to establish their effectiveness and security profile as well as the optimization of the physical-chemical and pharmacological performance. In clinical studies, most promising compounds are examined for their therapeutic validity for wide variety of conditions such as narcolepsy, cognitive disorders, obesity, schizophrenia, pain and addiction.

Osnovni cilj ovog istraživanja bio je utvrditi promjene kemijskih parametara svježine (histamina, trimetilamina i dimetilamina) lubina Dicentrarchus labrax tijekom skladištenja na različitim temperaturama (4±1°C i 12 ±1°C ). Pored toga, uspoređena je učinkovitost polusatnog tretmana ribe kupkom s 0, 9% NaCl u odnosu na klasično pakiranje ribe u podloške direktno s leda. Također je uspoređen i učinak dvije najčešće korištene vrste stiropornih podložaka na kemijske pokazatelje svježine ribe.

Biogenic amines are organic nitrogenous compounds produced in wines mainly during malolactic fermentation (MLF). Its presence is a health risk and it could negatively aff ect the wine quality. Th e objective of this work was to determine the biogenic amines content in Croatian Zweigelt wines produced with diff erent MLF inoculation time. Biogenic amines were determined by liquid chromatographic method. Results showed signifi cant infl uence of MLF on the concentrations of biogenic amines. Control wines had the lowest amount of total biogenic amines while wines in which MLF was conducted aft er alcoholic fermentation had signifi cantly the highest concentrations of total biogenic amines. Between them histamine was the most abundant amine ranging from 1.14 mg/L in control wines up to 2.94 mg/L in wines that undergo MLF aft er alcoholic fermentation. It can be concluded that MLF process and time of inoculation can signifi cantly infl uence the formation of biogenic amines. Th ese results suggest that co-inoculation using Oenoccocus oeni commercial strain Uvaferm ß is a worthwhile alternative compared to traditional post AF inoculation for Zweigelt winemaking.

Summary In recent years, there has been a steady increase in the prevalence of allergic diseases. Allergic immune response represents a complex network of cellular events involving numerous immune cells and mediators. It represents the interaction of innate and acquired immune response. The key role in the immune cascade is taken by histamine, a natural component of the body, which in the allergic inflammatory response is releasesd by the mast cells and basophils. The aim of this study was to highlight the role of histamine in allergic immunological events, their effect on Th1 and Th2 subpopulation of lymphocytes and the production of the corresponding cytokines, as well as the role of histamine blockers in the treatment of these conditions. Histamine achieves its effect by binding to the four types of its receptors, which are widely distributed in the body. Histamine blockers block a numerous effects of histamine by binding to these receptors. As a highly selective second-generation antihistamine, cetirizine not only achieves its effects by binding to H1 receptors, but also attenuates numerous events during the inflammatory process. Knowledge of the effects of histamine blockers, including cetirizine, may lead to the selection of proper therapy for the treatment of allegic diseases.

Standardi sigurnosti hrane zahtijevaju detekciju spojeva koji, čak i u tragovima, mogu utjecati na zdravlje ljudi. U ovu skupinu tvari pripadaju i biogeni amini, a jedan od najproučavanijih je histamin. Histamin se rijetko nalazi u svježoj ribi, međutim njegov udio raste s razvojem procesa bakterijske razgradnje ribe, te se histamin može smatrati indikatorom kvarenja ribe. Mariniranje je jedan od načina prerade ribljeg mesa kojim se, zahvaljujući djelovanju kombinacije octene kiseline i soli te drugih sastojaka marinade, zadržava djelovanje bakterija i enzima. Iako se snažan inhibitorski učinak marinade na bakterije i enzime povećava s povećanjem koncentracije, razvoj histamina moguć je i u mariniranoj ribi. Postoji više metoda za identifikaciju i kvantifikaciju odnosno određivanje udjela histamina u ribi. U ovom radu ispitivana je primjena Raman spektroskopije tj. spektroskopije površinski pojačanog Ramanovog raspršenja (SERS metode) i referentne HPLC metode kod određivanja udjela histamina u uzorcima marinirane ribe, te je izvršena usporedba dobivenih rezultata i podudarnosti korištenih metoda. Usporedba rezultata SERS metode i referentne HPLC metode pokazala je da je najveća točnost postignuta primjenom modela dobivenog linearnom regresijom na temelju intenziteta SERS vrpce histamina na 1264 cm-1i 1570 cm-1u rasponu koncentracija 0 - 200 mg kg-1. SERS metoda omogućava analizu histamina u uzorcima ribe uz značajno smanjene troškove te skraćuje vrijeme potrebno za analizu (30 min) u usporedbi s referentnom HPLC metodom (140 min). Food safety standards require detection of compounds that even at trace levels, may affect human health. This group of substances also includes the biogenic amines, and one of the most prominent is histamine. Histamine is rarely found in fresh fish but its share is growing with the development of bacterial degradation, thus histamine can be considered an indicator of fish degradation. Marinating is one method of fish processing, which, by combination of acetic acid, salt and other marinade ingredients keeps under control the bacteria and enzymes activity. Although the potent inhibitory effect of marinades on bacteria and enzymes increases with increasing concentrations, histamine development is also possible in marinated fish.There are several methods for identifying and quantifying histamine in fish.This thesis investigated the application of Raman spectroscopy apropos Surface Enhanced Raman Scattering (SERS method) and reference HPLC method to determine the histamine content in marinated fish samples, and compared the obtained results and the correlation of the methods used. Results of the SERS method and the reference HPLC method showed that the highest accuracy was achieved by applying a linear model regression based on the intensity of the SERS band of histamine at 1264 cm-1and 1570 cm-1 in the range of 0 - 200 mg kg-1. The SERS method allows analysis of histamine in fish samples at significantly reduced costs and shortens the time required for analysis (30 min) compared to the reference HPLC method (140 min).

Histamin je jedan od najproučavanijih biogenih amina ponajprije zbog svoje toksičnosti. Veoma je bitno na vrijeme detektirati histamin u ribi i proizvodima od ribe kako bi se na vrijeme spriječilo histaminsko trovanje. Zbog toga se nastoje razviti nove, brže metode za detekciju histamina, a jedna od tih metoda je i Raman spektroskopija. U ovom radu prikazana je usporedba postupka analize histamina primjenom tekućinske kromatografije visoke djelotvornosti (HPLC) i spektroskopije površinski pojačanog Ramanovog raspršenja (SERS). Utvrđeno je da SERS metoda ima neke prednosti pred HPLC metodom, a to su značajno pojednostavljenje postupka analize, skraćivanje vremena potrebnog za analizu jednog uzorka i smanjenje troškova analize. Usporedbom koeficijenta korelacije (R2) kalibracijskih modela temeljenih na intenzitetu vrpce na 1264 cm-1 (0.9784) i na 1570 cm-1 (0,9273), s koeficijentom korelacije standardnog modela dobivenog HPLC mjerenjem (0,995) vidljivo je da model temeljen na intenzitetu vrpce na 1264 cm-1 ne odudara puno od standardnog modela te može biti korišten za analizu i kvantifikaciju histamina. Histamine is one of the most investigated biogenic amines, primarily beacuse of its toxicity. It is very important to detect histamin early in fish and fish products to prevent histamin poisoning on time. This is why new methods are being developed to detect histamine, and Raman spectroscopy is one of those methods. In this study is presented a comparison of the histamine analysis using the reference HPLC method and SERS method. It has been established that the SERS method has some advantages over HPLC methode, which is a significant simplification of the analitycal procedure, shortening the time required for single sample analysis and reducing the cost of the analysis. By comparing the correlation coefficient (R2) of the calibration models based on intensity of histamine band at 1264 cm-1 (0,9784) and at 1570 cm-1 (0,9273) with the correlation coefficient of the standard model obtained by HPLC method (0,995) shows that the model based on intensity of histamine band at 1264 cm-1 does not differ much from the standard model and can be used for analysis and quantification of histamine

Histamin je najvažniji biogeni amin koji pri povišenoj koncentraciji u hrani morskog podrijetla predstavlja toksikološki rizik. Ramanova spektroskopija se pokazala brzom i invazivnom metodom za detekciju histamina, dok su tradicionalne metode skuplje i složenije te teže primjenjive u industrijskim uvjetima. Cilj ovog istraživanja bio je pročistiti ekstrakte različitim otapalima kako bi se histamin izdvojio iz vodenog sloja i bolje detektirao SERS (površinski pojačano Ramanovo raspršenje) metodom, te time izbjeći dugi postupak uparavanja i skratiti navedenu metodu. Koeficijent koleracije (R2= 0,9614) i intenzitet vrpce histamina na 1570 cm-1 pokazali su se najboljima kod ekstrakta B1 (ekstrakcija HCl-om), dok je kod ekstrakta B2 (ekstrakcija s izooktanom i vodom) najveća točnost metode (95,45%) kod vrpce histamina na 1264 cm-1. Zbog otežanih uvjeta u radu s ekstraktima B i C (ekstrakcija 0.1 i 0.5M NaOH), te tijekom dodavanja i miješanja isptivanih otapala (HCl, izooktan, cikloheksan), može se zaključiti da te postupke nije preporučljivo primjenjivati. Histamine is the most important biogenic amine which represents a toxicological risk at elevated concentrations in sea food. Raman spectroscopy proved to be a fast and invasive histamine detection method, while traditional methods are more expensive, complex and difficult to apply in industrial conditions. The aim of this study was to purify the extracts with different solvents in order to separate histamine from the aqueous layer and enable better detection by the SERS (Surface Enhanced Raman Scattering) method. Thus the relatively long evaporation process was to be avoided and the method would shorten. The correlation coefficient c (R2= 0.9614) and the histamine band intensity at 1570 cm--1 were the best for analysing extract B1 (extraction with HCl). The highest method accuracy (95.45%) in extract B2 (extraction with isooctane and water) was achieved by the histamine band at 1264 cm-1. Due to difficulties in analysing extracts B and C (extraction with 0.1 and 0.5 M NaOH), and during the addition and mixing with investigated solvents (HCl, isooctane and cyclohexane) such procedures are not recommended for use.

Histamin je najvažniji biogeni amin koji uzrokuje trovanje kod ljudi, a u organizam se unosi konzumacijom proizvoda ribarstva. Tradicionalne instrumentalne metode za detekciju histamina nisu adekvatne za primjenu u industriji pa se iz tog razloga istražuju nove metode poput Raman spektroskopije kako bi se osigurala kvaliteta i zdravstvena ispravnost proizvoda ribarstva. U ovom istraživanju, određivan je udio histamina u tkivu palamide s ciljem validacije razvijene SERS metode (spektroskopija površinski pojačanog Ramanovog raspršenja). Kao referentna metoda uzima se HPLC (tekućinska kromatografija visoke djelotvornosti). Snimljeni SERS spektri su obrađeni radi uklanjanja interferencija te su histaminske vrpce vidljive na 1264 i 1570 cm-1. Koeficijent korelacije (R2) za kalibracijski model koji se temelji na intenzitetu vrpce na 1264 cm-1 iznosi 0,951, a za model na 1570 cm-1 iznosi 0,990 te se isti pokazao kao bolji model. Histamine is the most important biogenic amine which is ingested by eating seafood products. It causes human intoxication. Traditional methods for analysis of histamine are not acceptable in industry use so new methods like Raman spectroscopy are being investigated to ensure quality and health standards of seafood products. In this study, histamine content in Atlantic bonito tissue has been determined to validate the developed SERS (Surface Enhanced Raman Scattering) method. HPLC (High Performance Liquid Chromatography) is the reference method. Recorded spectra have been processed to remove interferences and histamine bands are observed at 1264 cm-1 and 1570 cm-1. Correlation coefficient (R2) for calibration model based on intensity of histamine band at 1264 cm-1 is 0,951 cm-1 and 0,990 for model at 1570 cm-1. Based on given results, the last one is determined to be the more accurate model.

WOS: 000430650600016, In the present study, 0.00% (control), 0.10%, 0.40%, 0.70%, 1.00% and 1.30% black cumin oil was added to rainbow trout feed and the fish were fed for 120 days. Following the feeding period, the harvested fish were stored in the refrigerator at +/- 2 degrees C, in Styrofoam boxes filled with ice, for 23 days. During the storage period, the effects of black cumin oil on ammonia and biogenic amine production in rainbow trout were examined. An increase was observed in the ammonia content during the storage period, with the most substantial increase observed in the control group and the least substantial increase observed in the 1.30% black cumin oil group. The biogenic amine level increased during the storage period. A significant increases was observed in the tyramine level during the storage period (

Bu çalışmada, erkek sıçanlarda merkezi olarak enjekte edilen histaminin erkek hipotalamik-hipofizel-gonadal aksis üzerindeki etkileri ve bu etkilere merkezi sinir sisteminde bulunan histaminin H1R, H2R ve H3/4R'lerinin aracılığının araştırılması hedeflendi. Çalışmada 77 adet erkek Sprague Dawley ırkı sıçan kullanıldı. Sevofluran anestezisi altında plazma kan örneklerinin toplanabilmesi için sıçanların sol femoral arterlerine katater yerleştirildi. Merkezi yolla ilaç mikroenjeksiyonları için ise sıçanların serebral yan ventriküllerine kılavuz kanül yerleştirildi. Plazma GnRH, LH, FSH ve testosteron seviyelerini belirlemek için ise uygun ilaç enjeksiyonlarından önce ve enjeksiyondan sonraki 20., 40. ve 60. dakikalarda femoral artere yerleştirilen kateterden toplam 500 µl'lik kan örnekleri toplandı. Histaminin erkek hipotalamik-hipofizel-gonadal aksis üzerindeki etkilerini göstermek amacıyla histamin 50 ve 100 nmol dozlarında merkezi olarak uygulandı. Serebral yan ventriküle 50 nmol dozda histamin uygulanması, istatistiksel olarak anlamlı olmayan bir seviyede olmakla birlikte GnRH, LH, FSH ve testosteron hormonlarının seviyelerinde artışlar oluşturdu. Histaminin 100 nmol'lük dozunun merkezi olarak enjekte edilmesi ise, GnRH, LH, FSH ve testosteron hormonlarının seviyelerinde istatistiksel olarak anlamlı (p

Aim: Plasma-based biomarkers of functional cardiovascular and hemodynamic status have been fundamental discoveries that propelled modern clinical cardiology. The aim of this work was to provide a review of catestatin – a novel cardiac biomarker that might provide higher sensitivity and specificity in pinpointing and detecting specific phases of cardiovascular pathology progression, with the emphasis on arterial blood pressure. Materials and methods: An extensive literature search has been performed using the MEDLINE (PubMed) database to retrieve data from high-impact clinical trials and basic studies. Results: Based on the analysis of thirty original research studies that were selected among 75 screened articles, catestatin, a 21-amino acid residue that is proteolytically cleaved from chromogranin A is a promising novel biomarker of arterial blood pressure. Its physiological relevance lies in the ability to inhibit catecholamine secretion from adrenal chromaffin cells. Preliminary in vivo studies performed in humans demonstrated that catestatin exhibited direct vasoactive effects through marked vasodilation of blood vessels, especially in women. Furthermore, plasma concentrations of catestatin were consistently decreased in hypertensive subjects. In rat models, catestatin promoted the augmented release of histamine from mast cells and thus supported vasodilatory action. In addition, chronic administration of catestatin in rats induced cardioprotection, decreased sympathetic drive and improved autonomic function in subjects with hypertensive cardiomyopathy. In acute coronary syndromes, human subjects with ST- elevation myocardial infarction (STEMI) and unstable angina pectoris (UAP) had significantly lower levels of plasma catestatin compared to control groups. Conclusion: Plasma catestatin levels are inversely correlated with hypertension phenotype, especially in the early stages of hypertension development. Available data suggests that catestatin might play an important pathophysiological role in early hypertension as well as other pleiotropic cardioprotective roles. All of this makes catestatin worthy of further examination, especially in the light of preventive and therapeutic clinical applications.

Zbog svoje izuzetne toksičnosti histamin je jedan od najproučavanijih biogenih amina. Kako bi se spriječilo histaminsko trovanje od velike važnosti je rana detekcija histamina. Samim time došlo je do sve intenzivnijeg razvoja novih metoda za ranu detekciju histamina. Jedna od njih je Raman spektroskopija. U ovom radu prikazana je usporedba HPLC metode i Raman spektroskopije na uzorcima slanih inćuna. HPLC metoda se koristila kao referentna metoda a uspoređeni su koeficijenti kalibracijskih modela temeljenih na intenzitetu vrpce na 1264 cm-1 i na 1570 cm-1 s koeficijentom korelacije standardnog modela dobivenog HPLC mjerenjem. Dobivenim rezultatima vidljivo je da model temeljen na intenzitetu vrpce na 1264 cm-1 nema velika odstupanja, te se ta metoda može koristiti za analizu i detekciju histamina. Histamine is one of the most investigated biogenic amines, primarily because of its toxicity. In order to prevent histamine poisoning, early detection of histamine is of great importance. There is an increasing need for the development of new methods for early detection of histamine. One of them is Raman spectroscopy. In this study is presented a comparison of the HPLC method and the Raman spectroscopy on salty anchovy samples. The HPLC method was used as a reference method. We compared coefficients of calibration models based on the intensity of the tape at 1264 cm-1 and at 1570 cm-1 with the correlation coefficient of the standard model obtained by HPLC measurement. The obtained results showed that the model based on the intensity of the tape at 1264 cm-1 does not have large deviations, and this method can be used for analysis and detection of histamine.

Budući da razvitak histamina u ribi predstavlja veliku opasnost za ljude koji ju konzumiraju, godinama su se nastojale razviti nove i brze metode za detekciju histamina, a među njima se posebno istakla metoda Raman spektroskopije. U ovom radu prikazana je usporedba spektroskopije površinski pojačanog Ramanovog raspršivanja (SERS) sa tekućinskom kromatografijom visoke djelotvornosti (HPLC). Nakon analize, histaminske vrpce su bile vidljive na 1264 i 1570 cm-1 te je na osnovu analitičkog prinosa, odnosno manjeg odstupanja od referentne HPLC metode zaključeno da je SERS metoda temeljena na intenzitetu vrpce na 1264 cm-1 bolja u odnosu na SERS metodu temeljenu na intenzitetu vrpce na 1570 cm-1. Također usporedbom RSD vrijednosti i odstupanja SERS i HPLC metode, uočeno je da je HPLC metoda točnija i preciznija, no rezultati SERS analize su prihvatljivi, te SERS metoda ne odudara puno od standardnog modela te može biti korištena za analizu i kvantifikaciju histamina. Given the fact that the development of histamine in the fish brings grave danger to the people who eat it, new and fast histamine detection methods were developed during the years, the most significant of which is Raman spectroscopy. This paper compares the procedures of Surface Enhanced Raman Spectroscopy (SERS) with High-performance liquid chromatography (HPLC). The analysis has shown that histamine ribbons were visible at 1264 and 1570 cm-1. Based on the analytical production i.e. smaller deviation from the referent HPLC method it is safe to conclude that SERS method based on the intensity of the ribbon visible at 1264 cm-1 is better than the SERS method based on the intensity of the ribbon visible at 1570 cm-1. Also, comparison between RSD values and the deviations of SERS and HPLC method has shown that HPLC method is more accurate and precise, but the results of SERS analysis are acceptable, as well. Thus, SERS method is not significantly different from the standard model and is free to be used for analysis and quantification of histamine.

U disertaciji je proučen katalitički mehanizam te supstratna specifičnost enzima MAO B i DAO koji su uključeni u katabolizam histamina. Naime, pokazano je kako je izmijenjena aktivnost i ekspresija enzima MAO B povezana s neurodegenerativnim bolestima poput Parkinsonove bolesti te neurološkim poremećajima, poput psihoze, depresije i impulzivnosti. S druge strane, izmijenjena aktivnost enzima DAO razlog je intolerancije na histamin, stanja koje je popraćeno neugodnim simptomima i probavnim problemima. Stoga, svrha ove disertacije je pouzdano definiranje katalitičkog mehanizma oba enzima te utvrđivanje determinanti specifičnosti primjenom najsuvremenijih metoda u računalnoj biokemiji. Rezultati disertacije mogli bi poslužiti kao temelj u racionalnom dizajnu lijekova za liječenje navedenih patoloških stanja. U radu su primijenjene računalne metode koje se koriste za proučavanje strukture, dinamike, selektivnosti i reaktivnosti enzima kao što su molekulsko-dinamičke simulacije, izračun Gibbsove energije vezanja, kvantno-mehanički računi u okviru klaster pristupa te metoda EVB. Molekulsko-dinamičke simulacije spomenutih enzima te izračun Gibbsove energije vezanja rezultirali su objašnjenjem supstratne specifičnosti enzima MAO B, dok su kvantno-mehanički računi u okviru klaster pristupa te metoda EVB dali kvantitativni uvid u katalitički mehanizam oba proučavana enzima. The scope of this dissertation is the elucidation of the catalytic mechanism and substrate specificity of MAO B and DAO enzymes involved in the catabolism of histamine. Altered expression and activity of MAO B is associated with neurodegenerative diseases such as Parkinson's disease, and neurological disorders such as psychosis, depression and impulsivity. On the other hand, the altered enzyme activity of DAO is the root cause for the histamine intolerance, a condition which is characterized by unpleasant symptoms and gastrointestinal problems. Therefore, the purpose of this dissertation is to establish the catalytic mechanism of both enzymes together with the determinants of their specificity using the most advanced methods in computational biochemistry such as molecular dynamics simulations, binding free energy and quantum mechanical calculations within the cluster approach and the empirical valence bond (EVB) simulations. Molecular dynamics simulations and the binding free energy calculations explain the substrate specificity of MAO B enzyme, while the quantum mechanical calculations together with the EVB simulations provide a quantitative insight into the catalytic mechanism of both enzymes.

Zbog svoje toksičnosti, histamin je jedan od najproučavanijih biogenih amina. Rana detekcija histamina u ribi i proizvodima od ribe nužna je kako bi se na vrijeme spriječilo histaminsko trovanje. Iz tog razloga istražuju se nove metode poput Raman spektroskopije. U ovom radu ispitana je mogućnost primjene različitih postupaka pripreme uzoraka s ciljem bržeg i jednostavnijeg provođenja SERS analize.Za što uspješniju ekstrakcijuhistamina korištena su različita otapala: perkloratna kiselina (Ekstrakt A), 0,1 M NaOH (Ekstrakt B) i 0,5 M NaOH (Ekstrakt C). Nakon snimanja i obrade SERS spektara vidljive su karakteristične vrpce histamina na 1264i 1570 cm-1.Koeficijent korelacije (R2) za kalibracijski model ekstrakta B temeljenog na intenzitetu vrpce na 1264 cm-1iznosi 0,698, a za model ekstrakta B na1570 cm-1iznosi 0,991,te se isti pokazao kao najbolji model. Također,najveća točnost postignuta je primjenom istog modela dobivenog linearnom regresijom na temelju intenziteta SERS vrpce histamina na 1570 cm-1.Iako se postupak pripreme ekstrakta B za detekciju histamina SERS analizom pokazao uspješnim te vrlo blizu standardnom postupku, njegova daljnja primjena nije preporučljiva zbog problema koji se javljaju u procesu homogenizacije uzorka. Histamine is one of the most investigated biogenic amines because of its toxicity. Early detection of histamine in fish and fish products is necessary to prevent histamine poisoning on time. For this reason new methods like Raman spectroscopy are being investigated. The aim of this study is to investigate the possibility of applying different sampling procedures to improve the SERS analysis. To increase extraction of histamine various solvents were used: perchloric acid (Extract A), 0.1 M NaOH (Extract B) and 0.5 M NaOH (Extract C). After capturing and treating the SERS spectrum, characteristic histamine bands are visible at 1264 and 1570 cm-1. Correlation coefficient (R2) for calibration model of extract B based on intensity of histamine band at 1264 cm-1 is 0,698 cm-1 and 0,991 for model of extract B at 1570 cm-1. Based on given results, the last one is determined to be the more accurate model.Also, the highest accuracy was achieved by applying the same model obtained by linear regression based on intensity of histamine at 1570 cm-1. The procedure of preparing extract B for histamine detection has proven to be successful and very close to the standard procedure but its further use is not recommended because of problems occurring in the homogenization process.

Žutoperjana tuna se zajedno sa svojom podoricom Scombridae smatra glavnim uzročnikom skombroidnih trovanja tj. otrovanja histaminom. Taj biogeni amin nastaje neprikladnim skladištenjem svježe ribe te se nakon formiranja ne može ukloniti niti jednom metodom toplinske obrade. Stoga je cilj ovog istraživanja validirati metodu detekcije histamina spektroskopijom površinski pojačanog Ramanovog raspršenja (SERS) na uzorku tune te time omogućiti njegovo jednostavno, brzo i precizno određivanje. Rezultati su pokazali da se tom metodom mogu postići zadovoljavajući no ne i vrlo precizni rezultati u odnosu na referentnu HPLC metodu. Ipak, ističu se brojne prednosti SERS metode koje prvenstveno uključuju značajno smanjenje troškova i vremena potrebnog za analizu. Yellowfin tuna along with her family Scombridae is considered as the main cause of scombroid poisoning, ie. histamine poisoning. That biogenic amine will appear during unproperly storage of fish and once it is formed, it can't be removed by any method of heat treatment. According to that, purpose of this research is to validate method for detection of histamine using surface enhanced Raman spectroscopy (SERS) on tuna samples and enable his simple, fast and precise determination. Results have shown that using this method it is possible to achieve satisfying results, but not so precise according to reference HPLC method. However, there are a lot of advantages of SERS method like significant lowering of cost and time necessary for analysis.

Pterjium, oküler yüzey bozukluğu olup kapak aralığına uyan bölgedebulber konjonktivanın fibro¬vasküler proliferasyonunun korneayı invaze etmesisonucu oluşan bir patolojidir. Pterjium, cerrahi sonrası nüksün yüksek oranlaraulaştığı bir hastalıktır. Günümüzde nüks oranlarını daha aşağılara çekmekamacıyla birçok yeni cerrahi teknik yanında büyümesini durdurmak içinantineoblastikler kullanılmaktadır. Çeşitli büyüme faktörleri ve mediatörlerinbazı neoplazmlar gibi pterjium büyümesi, invazyonu ve nüksünde etkili olduğubulunmuştur. Kök hücre faktörü ve VEGF gibi büyüme faktörlerinin yanında,histaminde, karbonik anhidraz aktivitesinde, eph B4 sayısında ve nestinsalınımında artış bulunurken, IGFBP3 salınımında ise azalma bulunmuştur.Pterjimun büyümesinin durdurulması ve nüksünün önlenmesi için halen çalışmalaryapılmaktadır. Pterjium is an ocular surface disorder and is a pathologic result ofcorneal invasion of the fibro-vascular proliferation of the bulb conjunctiva inthe region corresponding to the lid space. Pterjium is a disease with highrates of recurrence after surgery. Nowadays, along with several new techniquesthat are used to prevent recurrence, antineoplastics are also used to stop thegrowth. Various growth factors and mediators, like some neoplasms, have beenfound to be effective in pterygium growth, invasion and relapse. In addition togrowth factors such as stem cell factor and VEGF, an increase in histamine,carbonic anhydrase activity, in number of eph B4, and in nestin release wasfound while a decrease in IGFBP3 release was found. Studies are currentlyunderway to stop the growth of pterygium and prevent recurrence.

Biogeni amini (BA) su skupina organskih dušikovih spojeva koji nastaju dekarboksilacijom aminokiselina. U ljudskom organizmu igraju ulogu moderatora u brojnim biološkim reakcijama koje mogu imati i neželjene posljedice. Određivanje biogenih amina u ribi važno je zbog mogućeg toksikološkog djelovanja na ljudski organizam nakon unosa velike količine biogenih amina te zbog činjenice da postoji veza između visokog udjela biogenih amina u ribi i loših higijenskih uvjeta tijekom proizvodnog procesa. U ovom radu ispitivana je mogućnost primjene Raman spektroskopije za analizu histamina u ribi i proizvodima od ribe. Prilikom analize realnih uzoraka histamin nije mogao biti detektiran zbog interferencija koje potječu od drugih tvari prisutnih u ribljem mišičju. Stoga su uzorci ribe pročišćeni te se postupkom pročišćavanja iz uzorka gubi oko 63 % ukupnih tvari uz gubitak od 38 % histamina čime se potvrđuje uspješnost pročišćavanja na smanjenje utjecaja matriksa prilikom analize Ramanovom spektroskopijom. Biogenic amines (BA) are the group of organic nitrogen compounds formed by decarboxylation of amino acids and their occurrence gives rise to significant debate of their physiological effects in humans. The study of biogenic amines in fish is important because the possible and the possibility that there might exist a relationship between hight amine contents and unsanitary conditions during the procedures. In this study the possibility of application of Raman spectroscopy for the analysis of histamine in fish and fish products. In the analysis of real samples histamine could be detected due to the interference originating from other substances present in fish muscles. Therefore, the fish samples purified and purification process of the sample loses about 63% of substances with a loss of 38% of histamine which confirms the success of treatment to reduce the impact of the matrix when analyzing Raman spectroscopy.toxicological risk that might results from an intake of a great quantity of these molecules.

Der GABA\(_{A}\)-Rezeptor (GABA\(_{A}\)R) ist der wichtigste inhibitorische Rezeptor im zentralen Nervensystem. Es handelt sich um einen Liganden-aktivierten pentameren Ionenkanal, der durch die \(\gamma\)-Aminobuttersäure aktiviert und durch eine Vielzahl von Substanzen, wie Benzodiazepine, Barbiturate, Alkohol, Neurosteroide, Kationen, Naturstoffe, Duftstoffe und dem Neurotransmitter Histamin, moduliert wird. Für viele dieser Substanzen sind die spezifischen Interaktionsstellen am GABA\(_{A}\)R bekannt, jedoch nicht für das Histamin. Um die Interaktionsstellen zu identifizieren wurden funktionale Analysen anhand von Chimären, welche aus der \(\beta\)3-Untereinheit des GABA\(_{A}\)R und der \(\alpha\)1-Untereinheit des Glycin-Rezeptors bestanden, durchgeführt. Darüber hinaus wurden 16 Aminosäuren hinsichtlich ihres Einflusses auf die modulatorische Wirkung von Histamin an dem GABA\(_{A}\)R durch gerichtete Mutagenese und funktionale Charakterisierung der mutierten Rezeptoren anhand der \(\textit {two-electrode Voltage-Clamp}\) Methode untersucht.

This bachelor thesis is focused on biogenic amines in fermented dairy products, particularly in cheeses. Biogenic amines are low molecular weight nitrogenous base formed mainly by decarboxylation of free amino acids by treatment with bacterial de-carboxylation enzymes. The most important bacteria forming biogenic amines in cheese are part of contaminating microflora (e.g. Enterococcus) and starter cultures (e.g. Lactobacillus). The cheeses are most prone to histamine, tyramine, cadaverine, putrescine. Furthermore, the work described effects, factors affecting the incidence and methods of determination.

Bu çalışmada, hemorajik şok oluşturulmuş sıçanlarda merkezi olarak enjekte edilen histaminin kardiyovasküler homeostaziste önemli rollere sahip beyin bölgeleri olan posterior hipotalamus (PH) ve nükleus traktus solitariustan (NTS) ekstraselüler asetilkolin (ACh) ve kolin (Ch) çıkışlarına etkisi ve bu etkide merkezi histaminerjik reseptörlerin aracılığının araştırılması amaçlandı. Çalışmada 140 adet erkek Wistar-Albino ırkı sıçan kullanıldı. Ketamin / ksilazin karışımı anestezisi altında kardiyovasküler parametrelerin kaydı ve hemorajik şokun oluşturulabilmesi için, sıçanların sol femoral arterlerine katater yerleştirildi. Merkezi yolla ilaç enjeksiyonları için ise sıçanların serebral yan ventriküllerine (s.y.v.) kılavuz kanül yerleştirildi. PH ve NTS'den ekstraselüler ACh ve Ch seviyelerini belirlemek amacıyla mikrodiyaliz çalışması yapıldı. Hemorajik şok, 10 dakikalık bir periyotta 100 gr vücut ağırlığı için 1,5 ml kan arteriyel katetater yoluyla alınarak oluşturuldu. Hemorajik şok, kan basıncında ve kalp atım sayısında şiddetli ve uzun süreli bir düşüşe ve bununla birlikte PH ve NTS ekstraselüler ACh ve Ch çıkışlarında artışlara neden oldu. Hemorajik şok sonrası uygulanan histamin (100 nmol) kardiyovasküler sistem üzerinde pressör ve taşikardik yanıt oluştururken, PH ve NTS ekstraselüler ACh ve Ch çıkışlarında hemorajiye bağlı olarak gelişen artışlara ilave artışlar da oluşturdu. Merkezi yolla uygulanan H1 reseptör antagonisti klorfeniramin ön tedavisi, hemorajik şok sonrası merkezi yolla uygulanan histaminin kardiyovasküler etkilerini ve PH veya NTS ekstraselüler ACh ve Ch çıkışları üzerindeki arttırıcı etkilerini tamamen bloke ederken, H2 reseptör antagonisti ranitidinin veya H3-4 reseptör antagonisti tioperamidin ön tedavisi ise histaminin oluşturduğu bu etkilerde herhangi bir değişikliğe sebep olmadı. Sonuç olarak elde edilen bulgular, hemorajik hipotansif koşullar altında merkezi olarak uygulanan histaminin, özellikle merkezi histaminerjik H1 reseptörlerini aktive ederek pressör ve taşikardik yanıtlar oluşturduğunu ve yine PH ve NTS ekstraselüler ACh ve Ch çıkışlarını artırdığını göstermektedir. The aim of this study was to determine the role of central administrated histamine in haemorrhagic hypotensive rats in the extracellular release of acetylcholine (ACh) and choline (Ch) from the posterior hypothalamus (PH) and nucleus tractus solitarius (NTS), the critical centres in the regulation of cardiovascular homeostasis as well as the mediation of central histamine receptors in the aforementioned parameter. 140 male Wistar-Albino rats were used in the study. In order to record cardiovascular parameters and to establish haemorrhagic shock, a catheter was inserted into the left femoral artery under mixture of ketamine and xylazine anesthesia. For central administration of drugs, a guide canula was directed into the lateral ventricle. To determine extracellular ACh and Ch levels from PH and NTS, microdialysis study was performed. Haemorrhagic shock was induced by withdrawing 1.5 ml blood per 100 gram body weight within a period of 10 minutes. Haemorrhagic shock provoked a severe and prolonged reduction in blood pressure and heart rate as well as an increase in the extracellular release of ACh and Ch from PH and NTS. Injection of histamine (100 nmol) after haemorrhagic shock caused an increase in blood pressure and tachycardic response, and restored hypotension. Meanwhile, it also provoked haemorrhage-dependent additional increase in extracellular levels of ACh and Ch from PH and NTS apart from normal increase. Central administration of H1 receptor antagonist chlorpheniramine pretreament completely blocked histamine induced pressor and tachycardiac cardiovascular responses and extracellular release of ACh and Ch from PH and NTS in haemorrhagic animals while, H2 receptor antagonist ranitidine or H3-4 receptor antagonist thioparamide pretreament did not block histamine induced cardiovascular responses.In conclusion, our findings show that centrally administered histamine under haemorrhagic hypotensive conditions induced pressor and tachycardiac reponses and at the same time caused the extracellular release of ACh and Ch from PH and NTS and restored hypotension particularly by activating H1 receptors. 111

TEZ12509 Tez (Doktora) -- Çukurova Üniversitesi, Adana, 2016. Kaynakça (s. 87-106) var. XV, 116 s. :_res. (bzs. rnk.), tablo ;_29 cm. Bu çalışmada, % 0.5 konsantrasyonda üç farklı organik asidin (propiyonik, asetik ve formik asit) iki farklı aminoasit (histidin ve tirosin) dekarboksilaz sıvısında referans bakteri (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Pseudomonas aeruginosa, Aeromonas hydrophila, Salmonella Paratyphi A, Campylobacter jejuni ve Yersinia enterocolitica) ve laktik asit bakterilerin (Lactococcus lactis subsp. lactis, Lactobacillus plantarum, Streptococcus thermophilus, Leuconostoc mesenteroides subsp. cremoris ve Pediococcus acidolactici) gelişimi ve biyojen amin üretimine etkisi incelenmiştir. Organik asitlerin bakterilere karşı minimium inhibisyon konsantrasyonu (MIK) ve minimum bakterisit konsantrasyonu (MBK) mikrodilüsyon yöntemine göre belirlenmiştir. Test edilen organik asitler arasında formik asit ve bunu takiben asetik asit diğer asitlere kıyasla daha yüksek antimikrobiyal aktivite sergilemiştir. Test edilen formik aside karşı en hassas türler 0.78 mg/mL MIK değeri ile E. coli, E. faecalis, S. Paratyphi A ve S. aureus olmuştur. Organik asitlerin referans bakteriler üzerindeki MBK değerleri genellikle 50 mg/mL’nin üzerinde olmuştur. Laktik asit bakterilerinde bu değer 6.52 mg/mL’ ye kadar düşmüştür. Aminoasit dekarboksilaz sıvısında genellikle en düşük bakteriyel yük formik asitte Pd. acidolactici bakterisinde (5.38 log kob/mL) gözlenmiştir. Bakteriler tarafından başlıca üretilen biyojen aminler dopamin, agmatin, putresin ve tiramin olmuştur. Histidin dekarboksilaz sıvısında histamin üretimi 0.12 mg/L (S. Paratyphi A) ve 28.56 mg/L (A. hydrophila) arasında değişkenlik gösterirken, laktik asit bakterileri tarafından üretilen en yüksek histamin değeri 2.38 mg/L (Lb. plantarum) olmuştur. Tirosin dekarboksilaz sıvısında en yüksek tiramin üretimi referans bakteriler için 578.22 mg/L (E. coli), laktik asit bakterileri için ise 22.05 mg/L (Lc. lactis) olmuştur. Organik asitlerin biyojen amin üretimindeki etkisi aminoasit dekarboksilaz sıvısına, asit tipine, bakteri türüne ve spesifik biyojen amine göre değişkenlik göstermiştir. In this study, the effects of tree different organic acids (propionic, acetic and formic acid) at doses of 0.5 % on growth and biogenic amine production of eight reference bacterial strains(Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Pseudomonas aeruginosa, Aeromonas hydrophila, Salmonella Paratyphi A, Campylobacter jejuni and Yersinia enterocolitica) and five lactic acid bacteria (Lactococcus lactic subsp. lactic, Lactobacillus plantarum, Streptococcus thermophilus, Leuconostoc mesenteroides subsp. cremoris and Pediococcus acidolactici) were investigated in two different amino acid (histidine and tyrosine) decarboxylase broth. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of organic acids against test strains were determined by using broth microdilution method. Among organic acids tested, formic acid showed the highest antimicrobial activity, followed by propionic acid. E. coli, E. faecalis, S. Paratyphi A and S. aureus were the most susceptible strains against formic acid, with MIC value of 0.78 mg/mL. MBC of reference strains was generally above 50 mg/mL, whilst lactic acid bacteria had 6.52 mg/mL of MBC. The lowest bacterial load in amino acid decarboxylase broth was observed by Ps. acidolactici (5.38 log cfu/mL) in the presence of formic acid. The main amines produced by bacteria were dopamine, agmatine, putrescine and tyramine. In histidine decarboxylase broth, histamine production was in range from 0.12 mg/L (S. Paratyphi A) to 28.56 mg/L (A. hydrophila), although the highest histamine production by lactic acid bacteria was found as 2.38 mg/L (Lb. plantarum). In tyrosine decarboxylase broth, the highest tyramine production was 578.22 mg/L for E. coli as references strains and 22.05 mg/L for Lac. subsp. lactis as lactic acid bacteria. The effect of organic acids on biogenic amine production varied depending on amino acid decarboxylase broth, type of acid, bacterial strains and specific biogenic amines. Bu çalışma Ç.Ü. Bilimsel Araştırma Projeleri Birimi tarafından desteklenmiştir. Proje No: FDK-2014-2526

Biogenic amines are low-molecular weight nitrogenic substances, which made by microorganisms with decarboxylic activity. These microrganisms contain decarboxylic enzymes, which catalyze decarboxylation of free amino acids. Biogenic amines are essential for organism, because they have their part in metabolism of celullar growth, regulativ of cardiac activity and so on. However, with higher concentrations ( > 100 mg/kg-1) they cause headache, hypotensia, nauzea, dizziness, and laminitiss with cattle. Laminitis is an inflammation of hooves. Goal of this work was so create an overview of origin, elimination and detection of biogenic amines in protein silages. The creation of biogenic amines is mainly caused by bad condition of silage making and its storage. Feeding of this silage can cause serious health issues to cattle and biogenic amines can be transferred to the animal products that could be potentionally harmfull for customers. Creation of these substances could be eliminated by adhering to the correct terms of silage making and its storage and by adding a correct inoculants. Detection of biogenic amides could be achived by chromatography.

Aminoasit dekarboksilasyonu sonucu oluşan biyojen aminler, hem vücudumuzda sentezlenmekte hem de gıdalarla alınabilmektedir. Meyve ve sebzelerde doğal olarak bulunmalarının yanında proteince zengin ve fermente edilmiş gıdaların olgunlaşması veya bozulması sonucu da ortaya çıkabilmektedirler. Gıdalarda aminoasitleri dekarboksilaz enzimi ile indirgeyen ve biyojen amin oluşumuna neden olan birçok mikroorganizma izole edilmiştir. Biyojen aminler, büyük ölçüde sucuk gibi et ürünleri yanısıra balık ürünleri, peynir, bira, şarap ve turşu gibi fermente gıdalar ve maya ekstraktları gibi pek çok üründe bulunmaktadır. Besin hijyeni ve sağlıklı beslenme bilincinin artmasıyla birlikte gıdalardaki biyojen amin varlığı gıda bozulması ve gıda güvenliği açısından tüketiciler için büyük önem taşımaya başlamıştır. Biyojen aminler gıdalarda iki nedenle önem arz etmektedirler: Bunlar biyojen amin miktarının kalite indikatörü olarak kabul ediliyor olması ve sağlık açısından riskli toksik etkilerinin bulunmasıdır. Bu çalışmada biyojen aminlerin oluşumu ve sağlığa etkileri incelenmiştir., Amino acid decarboxylation caused by biogenic amines, as well as to synthesize in our bodies and can be taken with food. Besides naturally present in fruits and vegetables protein-rich food and maturation of the fermented or degradation as a result may occur. Many microorganisms that cause reducing amino acid with decarboxylase enzyme and formation of biogenic amine was isolated in foods. Biogenic amines, largely meat products such as sausages and fish products, including cheese, beer, wine and fermented foods, pickles, yeast extract and are present in many products. Due to the increase of conciousness of food hygiene and healthy nutrition, the presence of biogenic amines in foods is increasing emphasis on both food security and food spoilage for consumers. Biogenic amines in foods are important for two reasons. These are the amount of biogenic amines is considered as quality indicators and the presence of risky toxic health effects. In this study the formation and the health effects of biogenic amines were investigated.