Subcellular localisation of the NME6 protein and anti-NME6 antibody testing

Nukleozid-difosfatske kinaze (NDPK/Nm23/NME) čine obitelj evolucijski očuvanih enzima, koji prenose fosfatnu skupinu γ s andenozin trifosfata na nukleozid difosfate i na taj način održavaju ravnotežu nukleotida u stanci, a smatra se da imaju i neke druge biokemijske funkcije. Nukleozid-difosfatske kinaze sudjeluju u brojnim biološkim procesima kao što su proliferacija, diferencijacija i razvoj, a imaju značajnu ulogu i u razvoju tumora i metastaziranju. Unatoč brojnim istraživanjima, njihova biološka funkcija nije do danas u potpunosti razjašnjena. Deset članova obitelji proteina NME (od NME1 do NME10) podijeljeni su u dvije skupine. Skupinu I čine proteini od NME1 do NME4 i svi posjeduju aktivnost kinaze NDP, dok u skupinu II ubrajamo NME5 do NME10, od kojih većina ne pokazuje aktivnost kinaze NDP. Strukturno, NME6 dijeli svega 34-41% homologije s proteinima iz prve skupine, dok je njegov izražaj relativno nizak u većini tkiva. Malobrojna istraživanja upućuju na to da je NME6 smješten u mitohondrijima i u citoplazmi. Cilj diplomskog rada bio je odrediti lokalizaciju proteina NME6 u humanim tumorskim stanicama HeLa uz pomoć fluorescentnog reporterskog sustava, testirati antitijelo anti-NME6 te optimizirati metodu Western blot. U tu svrhu, koristili smo metode transformacije, transfekcije, fluorescentnu imunocitokemiju, analizu Western blot te konfokalnu mikroskopiju.Nucleoside diphosphate kinases are evolutionary conserved enzymes which transfer the γ phosphate from adenosine triphosphates to nucleoside diphosphates consequently maintaining the nucleotide homeostasis in the cell. Additionally, they are also considered to have other biochemical functions. NDP kinases are involved in several biological processes such as proliferation, differentiation and development. Moreover, they are proven to have a significant role in tumour development and metastasis. However, despite intensive research in the field, all of their biological functions still aren’t completely resolved. The NME family consists of 10 genes/proteins (from NME1 to NME10), divided into two groups. Group I encompasses proteins from NME1 to NME4 which all exhibit the NDP kinase. Group II consists of proteins from NME5 to NME10 most of which show no NDP kinase activity. Structurally, NME6 shares only 34-41% homology with Group I proteins, while its expression is relatively low in most tissues. Scarce data on NME6 indicates that it is located in the mitochondria and the cytoplasm. The aim of this thesis was to determine the localization of the NME6 protein in HeLa tumour cells using GFP reporter system, to test the anti-NME6 antibody, and optimise the Western blot method. For that purpose, we used transformation, transfection and fluorescent immunocytochemistry methods, Western blot analysis and confocal microscopy

SAFETY ASPECTS OF BEE POLLEN USE IN NUTRITION

Bee pollen is popular nutraceutical and remedy used in traditional medicine since ancient times. Although it has indisputable beneficial action on human health, in recent years some issues regarding its safety have been raised. Mainly, they are a result of human actions, either indirectly (usage of pesticides, pollution with toxic trace elements) or directly (microbial contamination during handling). This review summarizes findings regarding safety aspects of bee pollen for human consumption

NUTRITION AS A SCIENTIFIC FIELD IN CROATIA: BIBLIOMETRIC ANALYSIS OF THE PERIOD 2010 – 2020

The scientific community’s interest in nutrition, i.e., the interaction of human and food consumption, is constantly rising worldwide and in Croatia. The published research results deepen our understanding of the processes involved in the interplay between food consumption and health, as well as different diseases and malfunctions in the human body. This research provides insight into scientific themes and focuses on nutrition research in Croatia. The study aims to provide an overview of the development of nutrition as a scientific field in Croatia from 2010 to 2020 using descriptive bibliometric analysis and science mapping. The analysis was made on a sample of 1173 articles indexed in the Web of Science Core Collection (WoS CC) database. In the analyzed period, nutrition and its impact on people\u27s health, especially on children\u27s health, are the focus of Croatian scientists\u27 research. Furthermore, it can be concluded that the same research trends continue in future research

COMMONLY USED ARTIFICIAL SWEETENERS IN EUROPE

It has been known for many years that the excessive consumption of sugar (sucrose) has harmful effects on human health. This fact led to a reduction in sugar consumption and the appearance of artificial sweeteners in the 1800s. The first low-cost and low-calorie sugar alternative was saccharin. Since this sweetener gained great popularity, other artificial sweeteners soon followed, including aspartame, acesulfame-K and cyclamates as the most common ones. As the result of a sharp rise in the obesity pandemic in all populations and ethnic groups, a demand for sweeteners with a minimum caloric value has increased dramatically in the last decade as consumers care more about their health. Due to the different regulation of permitted artificial sweeteners in United States (US) and Europe (EU), there are some controversies and suspicions about the relationship between certain sweeteners and a potential health risk. Despite doubts about the safety of artificial sweeteners, many studies have shown the absence of dangers associated with their use (if used in the acceptable daily intake, ADI). Therefore, artificial sweeteners today are considered as safe for consumption by many competent institutions and organisations. Nowadays, artificial sweeteners are fundamental in the food industry and present in many foodstuffs

Pathobiology of T-lymphoma

T–limfomi su tumori nastali od vlastitih izmijenjenih T–limfocita. Tipovi T–limfoma se međusobno teško razlikuju ukoliko se ne klasificiraju pomoću više, npr. razlike u morfološkoj strukturi zahvaćenog tkiva, imunofenotipu i histokemijske razlike. Poremećen regulatorni mehanizam rekombinacije VDJ gena je najvjerojatnije jedan od prvih i glavnih uzročnika razvoja T–limfoma, osim u malom broju slučajeva kada nastane funkcionalni triplet koji donosi veću varijabilanost TCR-a. U većini slučajeva nastane nefunkcionalni triplet ili stop kodon. T–limfome najčešće karakteriziraju translokacije, no mogu nastati i zbog delecije, ili mutacije u genomu čija je posljedica promijenjena ekspresija panT antigena. Takve stanice su nefunkcionalne i ukoliko ne nastupi programirana stanična smrt nastaju T–limfomi. Smatra se da virusi, npr. EBV, HIV i HTLV1, imaju značajan utjecaj na nastanak T–limfoma. Uočena je povezanost u učestalosti T–limfoma i geografskog položaja gdje je pojedini tip virusa zastupljeniji. Uzevši u obzir crnu, bijelu i žutu rasu, tipovi T–limfoma su različito zastupljeni, što se povezuje sa genetskim sličnostima unutar rase, odnosno razlikama između rasa.T–cell lymphoma is a type of cancer developed from it's own altered T-lymphocytes. Types of T–cell lymphoma are difficult to differentiate unless they are classified using multiple properties, e.g. differences in morphological structure of the affected tissue, immunophenotype and histochemical differences. Disrupted regulatory mechanism of VDJ genes recombination is probably one of the first and major causes of the T–cell lymphoma development. In rare cases the cause is the occurrence of a functional triplet, which brings in higher variability of the TCR, instead of a non-functional triplet or a stop codon. T–cell lymphoma is most often characterised by translocations, but it can be also developed by deletion or genome mutation resulting in changed expression of the panT antigen. These kind of cells are non-functional and if a programmed cellular death does not happen, the T–cell lymphomas develop. It is considered that viruses, e.g. EBV, HTLV1 and HIV have significant influence on the T–cell lymphoma formation. A correlation between the frequency of T–cell lymphoma appearance and geographic location is also perceived. Considering black, white and Asian races, T–cell lymphoma types have different occurrence that is linked to genetic similarities within a particular race as well as to differences between those races

Cultivation of tunicates in Adriatic Sea

Mali se broj ljudi bavi uzgojem plaštenjaka (Tunicata). Cilj ovog seminara je promocija i upoznavanje ove egzotične skupine životinja. U Sredozemlju su već zabilježena neka uzgajališta,te na temelju njihovih iskustava, predložen je način uzgoja u Jadranskom moru.Na kraju ćemo vidjeti mogućnosti plaštenjaka za proizvodnju hrane,lijekova te životinjske celuloze.A small number of people are engaged in tunicate farming (Tunicata). The objective of this seminar is to promote and get acquainted with this exotic animal group. Some ponds have already been recorded in the Mediterranean, and according to their experiences,a method of breeding is suggested in the Adriatic Sea. In the end we will see the possibility of producing food, medicine and animal cellulose from tunicates

Depositional environments and foraminiferal assemblages in sediments of the Neretva river delta plain

Na jezgrama NER3 i NER4 te iz rezultata preuzetih iz disertacije Felja (2017) za jezgre NER5 i NER6 iz područja deltne ravnice rijeke Neretve provedena je foraminiferska analiza, s ciljem da se rekonstruiraju taložni okoliši koji su se izmjenjivali tijekom holocena. U jezgrama je određena apsolutna brojnost foraminifera, relativna brojnost foraminifera i indeksi bioraznolikosti (indeks dominacije, Berger-Parker-ov indeks, Simpson-ov indeks, Shannon-Wiener-ov indeks, Fisher-ov indeks). Najzastupljenije vrste foraminifera pripadale su rodovima Ammonia, Haynesina i Porosononion. Povećanjem dubine rasla je i raznolikost vrsti, pa su nađene miliolide i jedinke roda Elphidium. Taložni okoliši koji su prepoznati na temelju indeksa bioraznolikosti i zastupljenosti foraminiferskih rodova su: aluvijalni okoliš (deltna ravnica), brakični okoliš (močvare/lagune, unutrašnji estuarij), te središnji/vanjski estuarij. Na temelju rezultata utvrđena je progradacija delte unutar estuarija te zapunjavanje estuarija tijekom holocena.On the cores NER3 and NER4, and from the results given in disertation Felja (2017) for cores NER5 and NER6 from the area of the Neretva river delta plain, a foraminiferal analysis was carried out to reconstruct the depositional environments that were changing during Holocene. In all drilled cores total number of foraminifera, relative percentage of foraminifera and biodiversity indices (Dominance, Berger-Parker index, Simpson index, Shannon-Wiener index, Fisher's index) were calculated. The most common types of foraminifera belonged to the genera Ammonia, Haynesina and Porosononion. The biodiversity increased with depth and various miliolids and Elphidium genus were present in the deeper layers. Depositional environments that were recognized based on biodiversity indices and the abundance of foraminiferal genera are: alluvial environments (deltaic plain), brackish environments (marhes/lagoons, inner estuaries), central/outer estuaries. Based on the results, progradation of intra-estuarine delta and the filling of the estuarine basin occurred during Holocene