A short review of headspace extraction and ultrasonic solvent extraction for honey volatiles fingerprinting

Honey volatiles exhibit a potential role in distinguishing honeys as a function of botanical origin, but heating of honey generates artefacts such as compounds of Strecker degradation and Maillard reaction products. This short review is focused on the most recently applied methods for honey volatiles fingerprinting (without generation of thermal artefacts): headspace extraction (dynamic headspace extraction (DHE), headspace solid-phase microextraction (HS-SPME)) and ultrasonic solvent extraction (USE). These methods display a varying degree of selectivity and effectiveness depending upon the compounds involved and the extraction conditions. Recent developments of these methods are discussed, with application examples drawn from the literature as well from our own research. Flavour qualities of the honey are very much dependent on the volatile and semivolatile organic compounds present in both the sample matrix and the headspace aroma. Therefore the use of one single technique is not adequate for reliable honey volatiles profiling, but combined use of headspace extraction and ultrasonic solvent extraction could be a useful tool for the characterization of the honey and identification of its botanical source through typical volatile marker compounds

Contribution to the Research of Free and Glycosidically Bound Volatiles from 2001 to 2006

U radu je prikazan doprinos istraživanju slobodnih i glikozidno vezanih isparljivih spojeva u razdoblju 2001. - 2006. koji je ostvaren radom u Zavodu za organsku kemiju Kemijsko-tehnološkog fakulteta, Sveučilište u Splitu. Identificirane su mnoge terpenske strukture, fenilpropanski derivati, kao i alifatski ugljikovodici, alkoholi, karbonili, masne kiseline i dr. Aglikoni oslobođeni enzimskom hidrolizom glikozida isparljivih spojeva bili su alifatski, fenilpropanski derivati, monoterpeni, seskviterpeni i dr. Nadalje, sintetizirani su glukozidi čestih monoterpenskih i alifatskih alkohola, te fenola (Koenigs-Knorrova glukozilacija, izravna glukozilacija uz FeCl3, enzimska kondenzacija i transglukozilacija iz celobioze).This review paper presents a contribution to the research of free and glycosidically bound volatiles in the 2001- 2006 period obtained by work in the Department of Organic Chemistry, Faculty of Chemistry and Technology, University of Split, Croatia. Many terpene structures, phenylpropane derivatives (Fig. 6), as well as aliphatic hydrocarbons, alcohols, carbonyls, fatty acids and others were identified (Fig. 5). The chemotypes of Origanum vulgare ssp. hirtum, Artemisia absinthium and Myrtus communis were determined and some uninvestigated or insufficiently investigated plants (Micromeria juliana, Sequaoiadendron giganteum, Populus nigra, Prunus mahaleb, Ailanthus altissima and others) were analyzed. A newly developed method of co-distillation enabled isolation of unstable volatile compounds without formation of artefacts (Fig. 1). The volatile aglycones liberated by enzymatic hydrolysis of the corresponding glycosides were aliphatic and phenylpropanoic derivatives, as well as monoterpenes, sesquiterpenes and others (Fig. 7). Only partial similarity was observed in the composition of volatile aglycones and corresponding free volatiles of the same plant. Furthermore, glucosides of ubiquitous monoterpene and aliphatic alcohols and phenols were prepared by Koenigs-Knorr glucosylation (Fig. 8), by enzymatic condensation and transglucosidation from cellobiose (Fig. 11) and by direct glucosylation with FeCl3 (Fig. 13). The products were characterized by GC-MS analysis of prepared tetraacetyl glucosides. Fragment ion characteristics of the aglucone moiety were present in all mass spectra, along with the fragments obtained from acetylated glucose (Fig. 12 and Fig. 14). Acetylated glucosides were separable on the HP-101 column, Fig. 9 (even diastereomeric tetraacetyl ß-glucosides of enantiomeric alcohols, Fig. 10)

HEADACHES IN CHILDREN

Uvod: Glavobolja je vrlo čest simptom u djece i adolescenata, a uzroci su raznoliki. Cilj: Raščlaniti i povezati anamnestičke podatke i nalaze djece hospitalizirane na Neuropedijatrijskom odjelu s dijagnozom glavobolje pri prijmu - povezati trajanje, kliničku sliku i uzroke glavobolja; oblik glavobolje klasificirati prema kriterijima The International Classification of Headache Disordersa, 2nd Edition. Metode: Retrospektivna analiza kliničkih značajki glavobolja u 377-ero djece praćene u razdoblju od 1. 1. 2003. Do 31. 12. 2007. Rezultati: Analiza obuhvaća podatke 221 (58.6%) djevojčice i 156 (41.4%) dječaka. Srednja dob ispitanika bila je 11godina i 8 mjeseci (raspon 2 godine i 8 mjeseci – 22 g.). Trajanje glavobolje ≤3 mj. Bilo je prisutno u 137-ero (36.3%) djece, u njih 220-ero (58.4%) >3mj, a u 20-ero (5.3%) djece podatci su bili nepoznati. Akutna glavobolja bila je prisutna u 144-ero (38.2%) djece, a recidivirajuća u njih 215-ero (57.0%). U sve djece učinjena je hematološko-biokemijsko-mikrobiološka, neurološka, okulistička i ORL obrada. Na temelju anamneze i ostalih nalaza glavobolje smo klasificirali kao: tenzijske (36.1%), moguće tenzijske (5.6%), povezane sa strukturnim promjenama organa glave (20.7%), migrena (16.7%), posttraumatske(3.2%), intrakranijske nevaskularne (neoplazme, disfunkcija drenažnog sustava prema Pudenzu) (1.8%) i vaskularne (CVI ) (0,3%). Učestalost glavobolja povezanih s infekcijom (sistemske infekcije, nakon preboljelog meningitisa) nađena je u 0.8% , zbog hipertenzije 0.5% slučajeva, dok su ostali uzroci (neklasificirane, sideropenična anemija, multipla skleroza, epilepsija, menstruacija) bili prisutni u 14.3% djece. Zaključak: Najčešći tip glavobolje bile su psihogene – tenzijske glavobolje.Radi utvrđivanja etiološke dijagnoze glavobolja koje zahtijevaju specifičnu terapiju, preporučujemo detaljnu kliničku obradu djece kojoj glavobolja traje dulje od tri mjeseca.Introduction: Headache is a common symptom in children and adolescents, with various causes. AIM: To divide and put together the anamnestic data and findings of children hospitalized at the Neuropaediatric Unit with headache as the primary diagnosis – to associate the duration, clinical features and causes of headaches and classify them according to the criteria of The International Classification of Headache Disorders Second Edition. Methods: The retrospective analysis of clinical data in 377 children followed up from January 1st 2003 until December 31st 2007. Results: The analysis includes data from 221(58.6%) girls and 156 (41.4%) boys. Mean age was 11 years (y) and 8 months (m) (2y8m – 22y). Headache with duration ≤3m was found in 137 (36.3%), >3m in 220 (58.4%) children. In 20 (5.3%) children the data was unknown. Acute headache was present in 144 (38.2%), recurrent headache in 215 (57.0%) children. In all children the haematological-biochemical-microbiological findings were made, together with neurological, ophthalmologic and ORL evaluation. Based on anamnestic data and findings we classified headaches as follows: tension-type (36.1%), possible tension-type (5.6%), associated with structural changes of the organs in the head (20.7%), migraine (16.7%), post-traumatic (3.2%), intracranial non-vascular (neoplasms, dysfunction of Pudenz drainage system) (1.8%) and vascular (CVI) (0,3%). Frequency of headaches associated with infection (systemic infection, after meningitis) was 0.8%, headache 0.5% was due to hypertension, while other causes were noted in 14.3% (non-classified, sideropenic anaemia, multiple sclerosis, epilepsy, menstruation). Conclusion: The most frequent type of headache was tension-type headache. In order to evaluate the etiological diagnosis of headaches which demand specific therapy, we recommend detailed clinical evaluation in children with duration of headaches of more than 3 months

GISAXS/GIXRD View of ZnO Films with Hierarchical Structural Elements

ZnO films constituted of porous sheet-like structures, formed by calcination of precursor, were examined using scanning electron microscopy and simultaneous small-angle scattering and diffraction of the synchrotron-sourced X-rays, under the grazing-incidence conditions. The presented analysis enabled insight into the complexity of the film morphology, which revealed substrate sensitivity on the microscopic and nanoscopic length scales. The average size and spatial arrangement of nanoparticles, single-crystal domains, and the average size and features of nanopores in sheet-like structures were determined for films deposited on glass, polycrystalline ZnO layer, and silicon

Functional dissection of the Sox9-Kcnj2 locus identifies nonessential and instructive roles of TAD architecture

The genome is organized in three-dimensional units called topologically associating domains (TADs), through a process dependent on the cooperative action of cohesin and the DNA-binding factor CTCF. Genomic rearrangements of TADs have been shown to cause gene misexpression and disease, but genome-wide depletion of CTCF has no drastic effects on transcription. Here, we investigate TAD function in vivo in mouse limb buds at the Sox9-Kcnj2 locus. We show that the removal of all major CTCF sites at the boundary and within the TAD resulted in a fusion of neighboring TADs, without major effects on gene expression. Gene misexpression and disease phenotypes, however, were achieved by redirecting regulatory activity through inversions and/or the repositioning of boundaries. Thus, TAD structures provide robustness and precision but are not essential for developmental gene regulation. Aberrant disease-related gene activation is not induced by a mere loss of insulation but requires CTCF-dependent redirection of enhancer-promoter contacts

Mineral and volatile composition of agua-mel from Portugal

Agua-mel (honey-water) is a typical honey-based product produced by the Portuguese beekeepers, particularly in southern Portugal. Agua-mel was characterized by mineral content and volatiles contents. Mineral content evaluation was performed based on a random sampling of 14 samples from a total of 16 samples provided by local producers. Mineral content showed that potassium predominated in agua-mel samples (1270-4105 mg/kg). The concentration of aluminium in one sample was tenfold higher (5.8 mg/kg) than in the remaining samples (0.3-0.6 mg/kg). Agua-mel volatiles were isolated by hydrodistillation and analysed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) from a subset of eight samples. Cluster analysis showed two poorly correlated clusters (S (corr) < 0.3). Cluster I only sample was dominated by trans-beta-ocimene (19 %), gamma-terpinene (15 %) and 2-furfural (9 %). Cluster II that included the remaining seven samples showed two moderately correlated subclusters (S (corr) < 0.5). The six samples with high correlation from subcluster IIa were dominated by 2-furfural (18-41 %) and benzene acetaldehyde (12-39 %). n-Nonadecane (14 %), n-heneicosane and 2-furfural (both 13 %) were the main components of subcluster IIb sample. Although the presence of some volatile compounds can help in the correlation between agua-mel and honey botanical source, the final product varies largely according to the preparation process even for the same producer, in different years. Agua-mel detailed characterization may assist in bringing added value to this typical Portuguese honey-based product

Psip1/p52 regulates posterior Hoxa genes through activation of lncRNA Hottip

Long noncoding RNAs (lncRNAs) have been implicated in various biological functions including the regulation of gene expression, however, the functionality of lncRNAs is not clearly understood and conflicting conclusions have often been reached when comparing different methods to investigate them. Moreover, little is known about the upstream regulation of lncRNAs. Here we show that the short isoform (p52) of a transcriptional co-activator—PC4 and SF2 interacting protein (Psip1), which is known to be involved in linking transcription to RNA processing, specifically regulates the expression of the lncRNA Hottip–located at the 5’ end of the Hoxa locus. Using both knockdown and knockout approaches we show that Hottip expression is required for activation of the 5’ Hoxa genes (Hoxa13 and Hoxa10/11) and for retaining Mll1 at the 5’ end of Hoxa. Moreover, we demonstrate that artificially inducing Hottip expression is sufficient to activate the 5’ Hoxa genes and that Hottip RNA binds to the 5’ end of Hoxa. By engineering premature transcription termination, we show that it is the Hottip lncRNA molecule itself, not just Hottip transcription that is required to maintains active expression of posterior Hox genes. Our data show a direct role for a lncRNA molecule in regulating the expression of developmentally-regulated mRNA genes in cis

Contribution to the Research of Free and Glycosidically Bound Volatiles from 2001 to 2006

U radu je prikazan doprinos istraživanju slobodnih i glikozidno vezanih isparljivih spojeva u razdoblju 2001. - 2006. koji je ostvaren radom u Zavodu za organsku kemiju Kemijsko-tehnološkog fakulteta, Sveučilište u Splitu. Identificirane su mnoge terpenske strukture, fenilpropanski derivati, kao i alifatski ugljikovodici, alkoholi, karbonili, masne kiseline i dr. Aglikoni oslobođeni enzimskom hidrolizom glikozida isparljivih spojeva bili su alifatski, fenilpropanski derivati, monoterpeni, seskviterpeni i dr. Nadalje, sintetizirani su glukozidi čestih monoterpenskih i alifatskih alkohola, te fenola (Koenigs-Knorrova glukozilacija, izravna glukozilacija uz FeCl3, enzimska kondenzacija i transglukozilacija iz celobioze).This review paper presents a contribution to the research of free and glycosidically bound volatiles in the 2001- 2006 period obtained by work in the Department of Organic Chemistry, Faculty of Chemistry and Technology, University of Split, Croatia. Many terpene structures, phenylpropane derivatives (Fig. 6), as well as aliphatic hydrocarbons, alcohols, carbonyls, fatty acids and others were identified (Fig. 5). The chemotypes of Origanum vulgare ssp. hirtum, Artemisia absinthium and Myrtus communis were determined and some uninvestigated or insufficiently investigated plants (Micromeria juliana, Sequaoiadendron giganteum, Populus nigra, Prunus mahaleb, Ailanthus altissima and others) were analyzed. A newly developed method of co-distillation enabled isolation of unstable volatile compounds without formation of artefacts (Fig. 1). The volatile aglycones liberated by enzymatic hydrolysis of the corresponding glycosides were aliphatic and phenylpropanoic derivatives, as well as monoterpenes, sesquiterpenes and others (Fig. 7). Only partial similarity was observed in the composition of volatile aglycones and corresponding free volatiles of the same plant. Furthermore, glucosides of ubiquitous monoterpene and aliphatic alcohols and phenols were prepared by Koenigs-Knorr glucosylation (Fig. 8), by enzymatic condensation and transglucosidation from cellobiose (Fig. 11) and by direct glucosylation with FeCl3 (Fig. 13). The products were characterized by GC-MS analysis of prepared tetraacetyl glucosides. Fragment ion characteristics of the aglucone moiety were present in all mass spectra, along with the fragments obtained from acetylated glucose (Fig. 12 and Fig. 14). Acetylated glucosides were separable on the HP-101 column, Fig. 9 (even diastereomeric tetraacetyl ß-glucosides of enantiomeric alcohols, Fig. 10)