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WOS:000584425500011 PubMed ID: 33281490 Research background. Utilization of wheat germ and wheat germ oil is limited due to high enzymatic activity and the presence of unsaturated fatty acids, which require stabilization techniques to overcome this problem. Experimental approach. In this study, the effects of stabilization methods (dry convective oven heating at 90 and 160 degrees C, microwave radiation at 180 and 360 W, and autoclave steaming) on both wheat germ and its oil were evaluated. Results and conclusions. Steaming caused the most dramatic changes in lipoxygenase activity, free fatty acid content, DPPH radical scavenging activity, and mass fractions of tocopherols and tocotrienols. Lower peroxide values were measured in the oil samples treated with convectional heating (160 degrees C) and steaming at temperatures above 100 degrees C. However, p-anisidine values of samples treated at higher temperatures were considerably greater than those of samples stabilized at lower temperatures. Oven heating at 160 degrees C was also one of the most effective treatments, after steaming, for the inactivation of lipoxygenase. Steaming significantly reduced mass fraction of total tocopherols, which was directly associated with the greater loss of beta-tocopherol content. On the contrary, gamma-and delta-tocopherol and tocotrienol homologues were abundant with higher amounts in steamed samples. alpha-Tocopherol and gamma-tocotrienol were the most resistant isomers to stabilization processes. Novelty and scientific contribution. This study shows that the high temperature oven heating method, which is widely used in the industry for thermal stabilization of wheat germ, does not provide an advantage in oxidative stability compared to steaming and microwave applications. Steaming delayed oxidation in the germ, while further inhibiting lipoxygenase activity. Moreover, tocotrienols were more conservable. In industrial application, low-power microwave (180 instead of 360 W) and oven heating at lower temperature (90 instead of 160 degrees C) would be preferable.

SUMMARY Common warts (verrucae vulgares) are the most common complaint in routine dermatological practice. Warts can be painful on pressure and are often an aesthetic problem, but they are not a major threat to the person’s general health. Treatment options are symptomatic and do not eradicate the causative agent. Dermatological surgery procedures such as cryotherapy, electrocauterization and excochleation can be painful, with common recurrences. These are the most important reasons for revival of the treatment procedures and remedies based on traditional medicine. Traditional medicine is still commonly practiced as a form of self-healing. This paper presents the most commonly used wart remedies of plant, animal and mineral origin, along with various magic practices. We emphasize that this paper is written from the viewpoint of physicians, practitioners of dermatology, not as a study in the history or culture. The main objective of the study was to explore various substances and methods people use as home remedies for warts. We performed a case study survey among the general population by interviewing 147 adult participants using a simple preliminary questionnaire inquiring about preferred treatment and knowledge about common warts.

SUMMARY The world is struggling to deal with the corona pandemic. Effective therapies are still awaited due to the lack of understanding of the pathophysiological mechanism of the disease. Bearing recent research and clinical observations in mind, the authors propose a novel physiological mechanism of COVID-19 and explain development of COVID-19 related acute respiratory distress syndrome (ARDS) secondary to COVID-19 related hemoglobinopathy. It is a consistent observation that the radiological picture of COVID-19 related ARDS bears more resemblance to high altitude pulmonary edema (HAPE) than typical ARDS. There has been great controversy regarding this proposed similarity. The main argument from those objecting to this comparison is that the etiology is hypoxia in case of HAPE and inflammation in COVID-19 related ARDS. We propose that considering the recent bioinformatics prediction models, COVID-19 might first infect red blood cells via CD147 and cause hemoglobin damage. The resulting hypoxemia may cause pulmonary hypoxic vasoconstriction leading to HAPE-like lung lesions. The now introduced alveolar hypoxia further exaggerates hemoglobinopathy hypoxemia leading to a vicious cycle. In this review, the authors recommend laboratory experiments to prove these hypotheses. The proposed physiological mechanism has significant therapeutic implications. If proven, the authors suggest the use of exchange transfusion as adjunct therapy and development of anti-CD147 drugs.

Robot-assisted brain tumor biopsy is becoming one of the most important innovative technologies in neurosurgical practice. The idea behind its engagement is to advance the safety and efficacy of the biopsy procedure, which is much in demand when planning the management of endocranial tumor pathology. Herein, we provide our earliest institutional experiences in utilizing this mesmerizing technology. Cranial robotic device was employed for stereotactic robot-assisted brain glioma biopsy in three consecutive patients from our series: an anaplastic isocitrate dehydrogenase (IDH) negative astrocytoma (WHO grade III) located in the right trigone region of the periventricular white matter; a low grade diffuse astrocytoma (WHO grade II) of bilateral thalamic region spreading into the right mesencephalic area; and an IDH-wildtype glioblastoma (WHO grade IV) of the right frontal lobe producing a contralateral midline shifting. Robot-assisted tumor biopsy was successfully performed to get tissue samples for histopathologic and immunohistochemical analysis. The adjacent tissue iatrogenic damage of the eloquent cortical areas was minimal, while the immediate postoperative recovery was satisfactory in all patients. In conclusion, considering the preliminary results of our early experiences, robot-assisted tumor biopsy was proven to be a feasible and accurate procedure when surgery for brain glioma was not an option. It may increase safety and precision, without expanding surgical time, being similarly effective when compared to standard stereotactic and manual biopsy. Using this method to provide accurate sampling for histopathologic and immunohistochemical analysis is a safe and easy way to determine management strategies and outcome of different types of brain glioma., Robotom potpomognuta tumorska biopsija postaje jednom od najvažnijih inovativnih tehnologija u neurokirurškom radu. Razlog njezine uporabe nalazi se u daljnjem poboljšanju sigurnosti, učinkovitosti i preciznosti biopsijske metode koja je osobito značajna u planiranju opskrbe endokranijske tumorske patologije. Ovim radom donosimo prva institucijska iskustva u primjeni ove začudne tehnologije pri biopsiji gliomskih tumora mozga. Kranijski robotički uređaj korišten je pri stereotaksijskoj robotom potpomognutoj tumorskoj biopsiji u tri susljedna slučaja iz naše serije, koja je uspješno učinjena radi uzimanja uzorka tumorskoga tkiva za patohistološku i imunohistokemijsku dijagnostiku u bolesnice s anaplastičkim izocitrat dehidrogenaza (IDH) negativnim astrocitomom (SZO st. III.) smještenim u periventrukulskoj bijeloj tvari desnoga trigonuma, u bolesnika s difuznim astrocitomom niskoga stupnja malignosti (SZO st. II.) smještenim obostrano u talamičkom području sa širenjem u desni mezencefalon, kao i u bolesnika s IDH-wildtype glioblastomom (SZO st. IV.) desnog čeonog režnja s pomakom središnjih tvorba. Jatrogena lezija pripadajućeg elokventnog moždanog korteksa bila je minimalna, dok je neposredni poslijeoperacijski oporavak bio uspješan u svih bolesnika. Uzimajući u obzir preliminarne rezultate našega početnog iskustva, zaključujemo kako je robotom potpomognuta tumorska biopsija dokazano izvodljiva i primjerena metoda u kirurgiji gliomskih tumora mozga kojom se može poboljšati sigurnost i preciznost bez produljenja vremena operacije, a koja je podjednako učinkovita u usporedbi sa standardnom stereotaksijskom i manualnom biopsijom. Uporaba navedene metode omogućuje precizno uzorkovanje tumorskoga tkiva za patohistološku i imunohistokemijsku analizu na siguran i lak način, što doprinosi odabiru strategije liječenja i predviđanju ishoda različitih tipova gliomskih tumora mozga.

Optimization of fermentation processes requires monitoring the species composition of starter cultures and their growth during fermentation. Most starter cultures contain closely related species. Nowadays, high-resolution melting (HRM) analysis is extensively used for multiplex identification of closely related species. In the present paper, we applied real-time polymerase chain reaction (PCR) with HRM analysis for the detection and differentiation of Lactobacillus sakei and L. curvatus. A primer pair was selected for the site of the rpoA gene of Lactobacillus spp. Eleven starter cultures and fifteen fermented sausages with a known bacterial composition were successfully tested using real-time PCR with HRM analysis with the developed primer pair.

Excessive intake of beer could increase serum uric acid levels, leading to high risk of gout, which was previously attributed to high purine content in beer. Recent reports that purine-rich vegetables and bean products do not cause higher uric acid levels do not support this theory. Why excessive intake of beer could increase a high risk of gout has been unclear. Other factors affecting the accumulation of uric acid in the blood have been explored. Beer contains relatively high levels of D-amino acids due to the racemization of L-amino acids induced by food processing. D-amino acid was catalyzed by D-amino acid oxidase to produce H2O2, which is further oxidized in the presence of Fe2+ to produce hydroxyl radicals, resulting in DNA damage and formation of a large amount of purine bases, which are oxidized to uric acid by a series of enzymes. Some food ingredients, such as vitamins and I–, prompt D-amino acids to form uric acid. D-amino acids in beer are one of the key factors responsible for the increase in uric acid levels. The biological response of D-amino acids could explain gout occurrence in beer drinkers.

Mezcal je destilat koji se proizvodi spontanom fermentacijom mošta dobivenog kuhanjem stabljika biljke Agave spp. i prešanjem dobivene smjese. Mošt agave sadržava velike količine fruktoze i fenolnih spojeva, a fermentacija se najčešće odvija pri stresnim i nekontroliranim uvjetima. Kvasci koji mogu rasti u takvim uvjetima obično imaju povoljna biološka i industrijska svojstva koja ih čine otpornijim, poput svojstva flokulacije. U ovom je radu sedam sojeva kvasca Saccharomyces cerevisiae izoliranih iz mošta agave izloženo temperaturama od 10 do 40 °C i uzgojeno u podlogama u prisutnosti fruktoze ili glukoze. Brojanjem kolonija u tzv. „microdrop“ testu potvrđeno je da su kvasci koji su rasli u podlozi s dodatkom fruktoze imali veću otpornost na stres uzrokovan niskom temperaturom (10 °C), u usporedbi s kvascima uzgojenim pri 40 °C. Soj kvasca koji je najbolje podnio stres (Sc3Y8) i komercijalni vinski kvasac (Fermichamp) uzgojeni su u tekućoj podlozi za fermentaciju i dugotrajno izloženi toplinskom stresu, radi određivanja njihove sposobnosti flokulacije. Tijekom fermentacije u podlozi s fruktozom povećalo se nakupljanje metabolita na kraju procesa, osobito pri 40 °C, te je proizvedeno 2,3 puta više glicerola (8,6 g/L), 1,3 puta više etanola (43,6 g/L) i 3,4 puta više octene kiseline (7,3 g/L) nego tijekom fermentacije u podlozi s glukozom. Pomoću konfokalnog mikroskopa utvrđene su morfološke promjene u kvascu, kao što su agregacija stanica i prisutnost ožiljaka na stijenci nastalih uslijed pupanja kvasca, osobito u soju Sc3Y8 izloženom temperaturi od 40 °C. Potvrđeno je da ovaj soj kvasca za proizvodnju pića mezcal flokulira u prisutnosti iona kalcija. Analizom ekspresije gena FLO1, FLO5 i FLO11 uključenih u regulaciju flokulacije potvrđeno je da je došlo do indukcije transkripcije u oba soja kvasca S. cerevisiae, naročito gena FLO5 u soju Sc3Y8., Mezcal is a distillate produced by spontaneous fermentation of the must obtained from stalks of Agave spp. plants that are cooked and pressed. Agave must contains a high amount of fructose and phenolic compounds, and fermentation usually occurs under stressful (and uncontrolled) environmental conditions. Yeasts capable of growing under such conditions usually display advantageous biological and industrial traits for stress tolerance such as flocculation. In this study, seven Saccharomyces cerevisiae strains isolated from mezcal must were exposed to temperatures ranging between 10 and 40 °C, and to different sugar sources (fructose or glucose). Yeasts grown in fructose increased their stress tolerance, determined by colony count in a microdrop assay, under low temperature (10 °C) compared to the growth at 40 °C on solid cultures. The most stress-tolerant mezcal strain (Sc3Y8) and a commercial wine (Fermichamp) strain, used as control, were grown under fermentation conditions and exposed to long-term temperature stress to determine their performance and their potential for flocculation. Compared to glucose, fermentation on fructose increased the metabolite accumulation at the end of culture, particularly at 40 °C, with 2.3, 1.3 and 3.4 times more glycerol (8.6 g/L), ethanol (43.6 g/L) and acetic acid (7.3 g/L), respectively. Using confocal microscopy analysis, we detected morphological changes such as aggregation and wall recognition at the level of budding scars in yeast, particularly in the Sc3Y8 strain when it was exposed to 40 °C. The analysis confirmed that this mezcal strain was positive for flocculation in the presence of Ca2+ ions. Analysis of FLO1, FLO5 and FLO11 gene expression implicated in flocculation in both Saccharomyces strains showed a strong transcriptional induction, mainly of the FLO5 gene in the mezcal Sc3Y8 strain.

Orally administered fungal vaccines show promise for the prevention of infectious diseases. Edible mushrooms are deemed appropriate hosts to produce oral vaccines due to their low production cost and low risk of gene contamination. However, their low expression level of antigens has limited the potential development of oral vaccines using mushrooms. The low expression level might result from impurity of the transgenic mycelia since dikaryotic mycelia are commonly used as transformation materials. In this study, stable transgenic hepatitis B virus surface antigen (HBsAg) in Flammulina velutipes transformants was obtained by Agrobacterium-mediated transformation, followed by fruiting and basidiospore mating. The formation of HBsAg was detected by western blot analysis. The expression levels of HBsAg in transgenic F. velutipes fruiting bodies were (129.3±15.1), (110.9±1.7) and (161.1±8.5) ng/g total soluble protein. However, the values may be underestimated due to incomplete protein extraction. Two of the four pigs in the experimental group produced positive anti-HBsAg-specific IgG after being fed the HBsAg transgenic F. velutipes fruiting bodies for 20 weeks, while no anti-HBsAg antibody was detected in the control group. One of the positive pigs had HBsAg titres of 5.36 and 14.9 mIU/mL in weeks 10 and 14, respectively, but expression faded thereafter. The other positive pig displayed HBsAg titres of 9.75, 17.86 and 39.87 mIU/mL in weeks 14, 18 and 20, respectively. The successful immunogenicity in pigs fed transgenic F. velutipes fruiting bodies demonstrated the potential of using the fungus as an oral vaccine.

Research background. During the current SARS-CoV2 pandemic, as well as earlier SARS and MERS epidemics, it has been observed that COVID19-positive women on average tend to have milder symptoms and lower fatality rates than men. There is a number of differences between the sexes known to contribute to different immune responses and severity of the disease, one being the effect of estrogen via estrogen receptor signalling. We wondered if estrogen might also affect the SARS-CoV2 more directly, perhaps by binding to the surface glycoprotein (S protein), thus possibly reducing its infectivity. Experimental approach. To assess whether there is a possibility for estrogen binding on the SARS-CoV2 S protein, we used BLAST and HHpred to compare protein sequences of S protein and human estrogen receptor β, while 3D structures of a potential estrogen binding site and an active site of estrogen receptor β were visualized and compared using PyMOL. Results and conclusions. By comparing the sequence of SARS-CoV2 S protein with the human estrogen receptor β, we identified a potential estrogen binding site on S protein and further determined that it also shares notable similarities with the active site of ER β when observed in 3D structure of their respective proteins. As a control, SARS-CoV2 S protein was compared with the human androgen receptor, and no such similarities were found. The potential estrogen binding site is part of coronavirus S2 superfamily domain, which is involved in host-virus membrane fusion during infection and appears to be conserved throughout the Coronaviridae family. Novelty and scientific contribution: This preliminary communication shows that SARS-CoV2 S protein features a potential estrogen binding site. Hopefully, this will prompt a more comprehensive study on the possibilities of estrogen binding on the S protein and the effect this might confer on the virus infectivity. © 2021, University of Zagreb. All rights reserved.

Objective: Halitosis, or oral malodour, is an unpleasant smell emanating from the oral cavity. It is a common complaint among patients with periodontitis, however, their relationship is not fully elucidated. This study aimed to evaluate the association between halitosis measures, clinical indicators of periodontitis and tongue coating, as well as a novel measure, periodontal inflamed surface area (PISA). Material and methods: Data of 10 patients with periodontitis and halitosis were included in this study. Halitosis was assessed by the organoleptic method and the portable sulphide monitor, measuring volatiles sulphur compounds. A comprehensive periodontal examination was conducted, and the parameters of probing depth, gingival recession, clinical attachment level, bleeding on probing, plaque and tongue coating were registered. The PISA was calculated using clinical attachment level, gingival recession and bleeding on probing. Results: A correlation between organoleptic score and tongue coating (r=0.554) and plaque (r=0.614) could be observed. No correlation between measures of halitosis and probing depth or the PISA could be detected. A significant correlation was found between organoleptic scores and volatiles sulphur compounds values (r=0.931). Conclusion: This pilot study has shown and further reiterated a complex interplay between different factors causative to halitosis in patients affected by periodontitis. The results suggest that tongue coating and oral hygiene may have an important role in halitosis in patients with periodontitis., Cilj istraživanja: Halitoza ili zadah neugodan je vonj koji se širi iz usne šupljine. Česta je pritužba među pacijentima s parodontitisom, no njihova povezanost nije u cijelosti razjašnjena. Cilj ovog istraživanja bio je procijeniti povezanost između parametara halitoze, kliničkih parametara parodontitisa i obloženosti jezika te novog parametra – površine parodonta zahvaćene upalom (engl. periodontal inflamed surface area – PISA). Materijali i metode: U ovo su istraživanje uključeni podatci 10 pacijenata s parodontitisom i halitozom. Halitoza je procijenjena organoleptičkom metodom i prijenosnim sulfidnim monitorom kojim se mjere hlapljive sumporne spojeve. Proveden je sveobuhvatni parodontni pregled i registrirani su parametri dubine sondiranja, recesija gingive, razine kliničkoga pričvrstka, krvarenja pri sondiranju, plaka i naslaga na jeziku. PISA je izračunata koristeći se razinom kliničkoga pričvrstka, recesije gingive i krvarenja pri sondiranju. Rezultati: Uočena je povezanost između organoleptičke ocjene halitoze i obloženosti jezika (r = 0,554) i plaka (r = 0,614). Nije zabilježena povezanost između parametara halitoze i dubine sondiranja ili PISA-e. Organoleptička ocjena halitoze i vrijednost hlapljivih sumpornih spojeva značajno su korelirali (r = 0,931). Zaključak: Ovo pilot-istraživanje pokazuje i ponovno potvrđuje složenu interakciju između različitih čimbenika koji uzrokuju halitozu kod pacijenata s dijagnozom parodontitisa. Rezultati sugeriraju da naslage na jeziku i oralna higijena mogu biti važni u slučaju halitoze kod pacijenata s parodontitisom.

SUMMARY The developments in transformation technology have enabled the scientists to incorporate, mutate or substitute gene(s) leading to a particular trait; advancing it to a point where only few technical limitations remain. Genotype dependency and explant types are important factors affecting transformation efficiency in potato. In the present study, a rapid, reproducible and stable Agrobacterium-mediated transformation procedure in potato was developed by a combination of different plant growth regulators. Leaf discs and internodal explants of five cultivars of potato, i.e. Lady Olympia, Granola, Agria, Désirée and Innovator were infected with Agrobacterium tumefaciens strain LBA4404 containing pBIN19 expression vector with β-glucuronidase gusA gene under the control of 35S CaMV promoter. Kanamycin was used as plant selectable marker for screening of primary transformants at concentration of 100 mg/L. Both explants responded positively; internode being more suitable explant for better transformation efficiency. Based on GUS histochemical assay, the transformation efficiency was 22, 20, 18.6, 15 and 10% using the internodal explant, and 15, 12, 17, 8 and 6% using leaf discs as explant in Lady Olympia, Granola, Agria, Désirée and Innovator respectively. Furthermore, PCR assays confirmed the presence of gusA and nptII genes in regenerated plants. The molecular analysis in succeeding progeny showed proper integration and expression of both genes. The results suggest Lady Olympia as the best cultivar for future transformation procedures. Overall, the short duration, rapidity and reproducibility makes this protocol suitable for wider application of transgenic potato plants.

SUMMARY The study focuses on predictive modelling of inactivation of Salmonella enterica after treatment with chlorophyllin-chitosan complex and visible light. Salmonella cells were incubated with chlorophyllin-chitosan complex (0.001% chlorophyllin and 0.1% chitosan) for different times (5-60 min) and then illuminated with visible light (λ=405 nm, He=38 J/cm2). Inactivation curves and post-treatment regrowth curves were built based on microbiological viability tests and data were fitted to ten inactivation and two regrowth models. The photoactivated complex reduced Salmonella population, which were unable to regrow. Weibull and Baranyi models were the best to describe the inactivation and regrowth kinetics respectively. In conclusion, data from the kinetic analysis and predictive modelling confirmed that photoactivated chlorophyllin-chitosan complex is a promising non-thermal approach for inactivation of Gram-negative pathogens, since no bacterial regrowth after treatment has been predicted.

The prevalence and importance of male infertility make it a serious worldwide medical and social problem. The aim of this study was to determine if there were any significant differences in zinc concentrations in seminal fluid in men diagnosed with infertility compared to control group, and if there were, to determine how it affected the number and percentage of sperm cells with normal motility and morphology. A case-control study was conducted in the Osijek-Baranja County from January 2014 to June of 2015. The study included 276 subjects referred to biochemistry laboratory of the Osijek University Hospital Centre, 100 (36.2%) of them with low sperm count confirmed by spermiogram after 3 months. Semen samples were processed according to the World Health Organization criteria, and zinc concentrations were determined using spectrophotometry and direct colorimetry without deproteinization on a chemistry analyzer (Olympus AU 680, Beckman Coulter, Tokyo, Japan). Study results showed the men with low sperm count to be significantly older (Mann Whitney U test: p=0.013) and to have lower zinc levels (χ2-test: p, Muška neplodnost s obzirom na njenu učestalost i važnost predstavlja ozbiljan globalni medicinski i socijalni problem. Cilj ovoga istraživanja bio je utvrditi postoje li značajne razlike koncentracije cinka u sjemenoj tekućini muškaraca s dijagnozom smanjene plodnosti u odnosu na kontrolnu skupinu te ako postoje, utvrditi kako razina cinka djeluje na broj, postotak pokretnih i broj morfološki normalnih spermija. Studija slučajeva i kontrola provedena je na području Osječko-baranjske županije u razdoblju od siječnja 2014. godine do lipnja 2015. godine. U studiju je bilo uključeno 276 ispitanika koji su bili upućeni u biokemijski laboratorij KBC-a Osijek, od kojih je kod 100 (36,2%) ispitanika potvrđena dijagnoza smanjene plodnosti ponovljenim spermogramom nakon 3 mjeseca. Uzorci sjemene tekućine obrađeni su u skladu s kriterijima Svjetske zdravstvene organizacije, a koncentracija cinka određena je spektrofotometrijskom, direktnom kolorimetrijskom metodom bez deproteinizacije na kemijskom analizatoru tipa Olympus AU 680 (Beckman Coulter, Tokyo, Japan). Istraživanje je pokazalo kako su muškarci s dijagnozom smanjene plodnosti značajno stariji (Mann Whitney U test: p=0,013) i imaju značajno niže vrijednosti cinka (χ2-test: p

Research background. Carob is widely cultivated Mediterranean plant, but its flour is scarcely used in bread-making. Previous studies investigated the quality of wheat bread with added carob flour showing discrepant results. This study aimed to investigate the fermentation performance, antioxidant activity, rheological behaviour, and baking application of carob sourdough. Experimental approach. Carob sourdough was fermented with Lactobacillus brevis or Lactobacillus fermentum combined with Saccharomyces cerevisiae for 24 h at 30 °C. At the end of sourdough fermentation number of viable lactic acid bacteria and yeast cells, titratable acidity, pH value, antioxidant activity, phenolics and sugar content of sourdough were determined. Carob flour (12 % of flour mass) or sourdough equivalent (22.5 % of dough mass) was applied in making composite partially baked frozen bread. Dough rheology was monitored using a farinograph. Rebaked bread samples were evaluated for nutritive value, physical properties, and sensory attributes using a hedonic test. Results and conclusions. By the end of fermentation, carob sourdough reached pH 4.2-4.5 and total acidity 9.9-12.3 mL of 0.1 M NaOH, sugar content was reduced for 8 g/100 g d.m, while total phenolics and antioxidant activity were increased up to 21 %, depending on the starter culture. Addition of carob flour or sourdough to wheat dough resulted in higher consistency, longer development time, and lower quality number. Regardless, bread with carob flour had significantly improved specific volume of bread. Compared with common wheat bread, carob breads had increased dietary fibre content (46 %), total phenolics (140-200 %) and antioxidant activity (240-300 %), higher shape, reduced crumbliness, unchanged firmness, and darker crumb colour. Consumers’ preference and overall acceptability scores of carob sour breads were comparable to those of wheat bread, falling into the category of 'liking very much'. Novelty and scientific contribution. To our knowledge, this is the first study that proved the feasibility of carob sourdough fermentation using mixed starter cultures, where L. brevis associated with S. cerevisiae was better adapted to the substrate than L. fermentum. The carob sourdough could be used as a natural ingredient for improvement of nutritive value and reduction of crumbliness of partially baked frozen bread.

The aim of this study was to determine plasma adropin concentration and parameters of insulin resistance in obese male type 2 diabetes mellitus (T2DM) patients before and after 3-month liraglutide treatment. In this interventional study, we enrolled 15 obese male T2DM patients with body mass index (BMI) >35 kg/m2, uncontrolled disease and HbA1c >7.5%, having previously taken taking two oral antidiabetic drugs. We modified their therapy to metformin and liraglutide for the next three months. After three months of liraglutide treatment, we observed significant decrease in body weight (from 111.5±18.7 kg to 109.2±17.5 kg, p=0.016) and BMI (from 40.9±7.3 to 40.1±7.0 kg/m2, p=0.021). Plasma adropin concentration increased significantly (p=0.003) compared with baseline. Fasting plasma insulin level decreased from 17.79±6.53 to 13.38±3.51 mU/L (p=0.002), fasting plasma glucose level decreased from 8.66±3.07 to 7.41±2.21 mmol/L (p=0.004) and HbA1c decreased from 7.98±0.70% to 7.26±0.36% (p=0.003). Insulin resistance presented as HOMA-IR decreased significantly from 7.30±5.19 to 4.52±2.61 (p=0.002). Systolic blood pressure, lipid status, liver and kidney function improved, but not reaching statistical significance. Treating obese male T2DM patients with liraglutide resulted in a significantly higher plasma adropin concentration, significant weight loss and improved parameters of insulin resistance, i.e. decreased fasting plasma insulin, plasma glucose levels and HOMA-IR., Cilj je bio usporediti plazmatske vrijednosti adropina i parametre inzulinske rezistencije kod pretilih muškaraca koji boluju od šećerne bolesti tip 2 (ŠBT2) prije i nakon 3 mjeseca primjene liraglutida. U ovoj intervencijskoj studiji sudjelovalo je 15 pretilih muškaraca koji boluju od ŠBT2 s indeksom tjelesne mase (ITM) >35 kg/m2, loše reguliranom bolešću i HbA1c >7,5%. Ispitanici su prethodno u terapiji imali dva peroralna antidijabetična lijeka. Nakon uključenja u studiju terapija im je modificirana na metformin i liraglutid tijekom tri mjeseca. Nakon primjene liraglutida kod ispitanika je zamijećeno smanjenje tjelesne mase (sa 111,5±18,7 na 109,2±17,5 kg, p=0,016) i ITM (s 40,9±7,3 na 40,1±7,0 kg/m2, p=0,021), dok su plazmatske vrijednosti adropina bile značajno povišene (p=0,003). Zamijećeno je sniženje vrijednosti inzulina natašte (sa 17,79±6,53 na 13,38±3,51 mU/L, p=0,002), glukoze natašte (s 8,66±3,07 na 7,41±2,21 mmol/L, p=0,004) te HbA1c (sa 7,98±0,70% na 7,26±0,36%, p=0,003). HOMA-IR se značajno smanjio (sa 7,30±5,19 na 4,52±2,61, p=0,002). Također su zabilježene niže vrijednosti sistoličkog arterijskog tlaka, bolji lipidni profil te poboljšanje jetrene i bubrežne funkcije, iako ne statistički značajno. Primjena liraglutida u pretilih muškaraca koji boluju od ŠBT2 rezultira statistički značajno višim razinama plazmatskog adropina, značajnim smanjenjem tjelesne težine i poboljšanjem svih parametara inzulinske rezistencije, tj. sniženjem plazmatskog inzulina i glukoze natašte te nižim HOMA-IR.

The role of T regulatory lymphocytes (Treg) particularly in cancer is well known. The goal of the present study was to determine the contribution of these lymphocytes in the regulation of anti-tumor immunity of CBA/HZgr mice against MC-2 fibrosarcoma (4th generation of methylcholanthrene induced tumor). The levels of T lymphocytes (CD4+, CD8+ and CD4+CD25+) were determined 8 and 20 days after tumor transplantation. Further, the role of CD4+CD25+ (Tregs) in tumor-host interaction was evaluated in vitro and in vivo by using specific monoclonal antibodies. We found that splenocytes of both control and Treg depleted tumor bearing mice strongly but differently inhibited growth of tumor cells in vitro. While splenocytes of untreated mice exhibited significant decrease of this activity (from 74.4% to 62.6% and 32.95%), the splenocytes of Treg depleted mice showed increase of this activity (from 79.5% to 84.3% and 86.2%) from day 6 to day 13 and day 21 after tumor grafting, respectively. Further, upon i.v. injecting specific monoclonal anti-Treg antibody tumor immediately prior to tumor cell intracutaneous transplantation, the tumor was rejected after initial growth. In treated mice, the incidence of Treg cells was very low initially, reaching normal values two weeks later. These animals were shown to be resistant to tumor transplantation four months later., Uloga T regulatornih limfocita dobro je poznata, osobito kod tumora. Cilj ovoga istraživanja bio je utvrditi doprinos ovih limfocita u regulaciji antitumorske imunosti CBA/HZgr miševa protiv MC-2 fibrosarkoma (4. generacija metilkolantrenom izazvanog tumora). Razine T limfocita (CD4+, CD8+ and CD4+CD25+) određivale su se 8 i 20 dana nakon transplantacije tumora. Nadalje, uloga CD4+CD25+ T limfocita (Treg) u interakciji tumora i domaćina procijenjena je in vitro i in vivo primjenom specifičnih monoklonskih protutijela. Utvrdili smo da splenociti kako kontrolnih miševa tako i miševa s tumorom bez Treg snažno, ali različito suzbijaju rast tumorskih stanica in vitro. Dok su splenociti netretiranih miševa pokazivali značajno smanjenje ove aktivnosti (sa 74,4% na 62,6% i 32,95%), splenociti miševa bez Treg pokazivali su porast ove aktivnosti (sa 79,5% na 84,3% i 86,2%) od 6. do 13. i 21. dana nakon transplantacije tumora. Uz to, nakon i.v. injektiranja specifičnog monoklonskog anti-Treg tumorskog protutijela neposredno prije intrakutane transplantacije tumorskih stanica, nakon početkog rasta nastupilo je odbacivanje tumora. Kod tretiranih miševa incidencija Treg stanica bila u početku vrlo niska i dostigla je normalne vrijednosti dva tjedna kasnije. Nakon četiri mjeseca pokazalo se da su ove životinje otporne na transplantaciju tumora.

SUMMARY Bone drilling causes focal temperature rise due to metal-to-bone contact, which may result in thermal osteonecrosis. Newly constructed internally cooled medical drill of an open type decreases temperature rise at a point of metal-to-bone contact although standard sterilization of such a drill could be inadequate due to bacteria retention within the drill lumen. The aim of this pilot study was to examine the effectiveness of sterilization and to propose sterilization recommendations for internally cooled open type bone drills. Unused internally cooled medical steel bone drills were tested. Drills were contaminated with Pseudomonas aeruginosa, Bacillus sp., beta-hemolytic Streptococcus sp., Enterobacter sp. and methicillin-resistant Staphylococcus pseudintermedius and then incubated for 24 hours at 37 °C. Afterwards, drills were autoclaved for 15, 20 and 30 minutes at 132 °C and 2.6 bar. When 15-minute sterilization was used, one out of 16 drills was contaminated with Pseudomonas aeruginosa, while the other 15 drills were sterile. Extended cycle sterilization in autoclave lasting for 20 and 30 minutes resulted in 100% sterility of all drills tested. In conclusion, lumened drills should be exposed to extended sterilization times in autoclave. Minimal recommended time for sterilization of lumened drills is 20 minutes.

Radi utvrđivanja utjecaja fermentacije na kakvoću proteina u koncentratu proteina graška ispitani su sljedeći parametri: udjel ukupnih fenola i tanina, aktivnost inhibitora proteaze, sastav aminokiselina i probavljivost proteina in vitro nakon 11 sati fermentacije s pomoću bakterije Lactobacillus plantarum. Maseni se udjel fenola u koncentratu proteina graška, izražen kao ekvivalent katehina, povećao na bazi suhe tvari s 2,5 pri 0 h na 4,9 mg/g nakon 11 sati fermentacije. Udjel tanina se povećao s 0,14 pri 0 h na maksimalnih 0,96 mg/g koncentrata nakon 5 h fermentacije, a zatim se smanjio na 0,79 mg/g nakon 11 h fermentacije. Nakon 9 h fermentacije smanjila se aktivnost inhibitora tripsina, međutim, pri svim ostalim vremenima fermentacije dobivene su vrijednosti slične onima pri 0 h. Aktivnost se inhibitora kimotripsina smanjila s 3,7 na 1,1 jedinicu inhibicije kimotripsina po mg nakon 11 sati fermentacije. Probavljivost je proteina dosegla maksimalnu vrijednost od 87,4 % nakon 5 sati fermentacije, međutim vrijednost aminokiselina koje sadržavaju sumpor smanjila se s 0,84 pri 0 h na 0,66 nakon 11 h fermentacije. Smanjenjem udjela sumpora promijenila se vrijednost aminokiselina korigirana probavljivošću proteina in vitro s 67,0 pri 0 h na 54,6 % nakon 11 h fermentacije. Dobiveni podaci potvrđuju da je, iako je fermentacija valjana metoda za smanjenje udjela nekih nenutritivnih sastojaka u koncentratu proteina graška, potrebno odabrati odgovarajuće bakterije koje nemaju izraženu sposobnost razgradnje aminokiselina što sadržavaju sumpor., In order to determine the impact of fermentation on protein quality, pea protein concentrate (PPC) was fermented with Lactobacillus plantarum for 11 h and total phenol and tannin contents, protease inhibitor activity, amino acid composition and in vitro protein digestibility were analyzed. Phenol levels, expressed as catechin equivalents (CE), increased on dry mass basis from 2.5 at 0 h to 4.9 mg CE per 1 g of PPC at 11 h. Tannin content rose from 0.14 at 0 h to a maximum of 0.96 mg CE per 1 g of PPC after 5 h, and thereafter declined to 0.79 mg/g after 11 h. After 9 h of fermentation trypsin inhibitor activity decreased, however, at all other fermentation times similar levels to the PPC at time 0 h were produced. Chymotrypsin inhibitor activity decreased from 3.7 to 1.1 chymotrypsin inhibitory units (CIU) per mg following 11 h of fermentation. Protein digestibility reached a maximum (87.4 %) after 5 h of fermentation, however, the sulfur amino acid score was reduced from 0.84 at 0 h to 0.66 at 11 h. This reduction in sulfur content altered the in vitro protein digestibility-corrected amino acid score from 67.0 % at 0 h to 54.6 % at 11 h. These data suggest that while fermentation is a viable method of reducing certain non-nutritive compounds in pea protein concentrate, selection of an alternative bacterium which metabolises sulfur amino acids to a lesser extent than L. plantarum should be considered.

Izoelektrična točka, duljina molekule, molekularna masa i slijed aminokiselina bitno utječu na topljivost proteina. U ovom smo se radu fokusirali na sastav aminokiselina i ispitali one koje najviše utječu na razinu ekspresije topljivog protutijela albumina iz ljudskog seruma (HSA dAb). Grupiranjem i primjenom linearnog modela analizirana je topljivost 65 varijanti proteina. Bitan utjecaj na ekspresiju topljivog protutijela dAb imale su specifične kombinacije aminokiselina, i to (S, R, N, D, Q) u supernatantu, (G, R, C, N, S) u lizatu peleta i (R, S, G) u ukupnom topljivom protutijelu dAb. Od 20 aminokiselina, arginin je imao negativan, a glicin i serin su imale pozitivan učinak na razinu ekspresije topljivog proteina. Preciznost linearnog modela predviđanja topljivosti proteina bila je 80 %. Zaključeno je da se povećanjem udjela polarnih aminokiselina, osobito glicina i serina, te smanjenjem udjela arginina bitno povećala ekspresija topljivog proteina HSA dAb., It is widely accepted that features such as pI, length, molecular mass and amino acid (AA) sequence have a significant influence on protein solubility. Here, we mainly focused on AA composition and explored those that most affected the soluble expression level of human serum albumin (HSA) domain antibody (dAb). The soluble expression and sequence of 65 dAb variants were analysed using clustering and linear modelling. Certain AAs significantly affected the soluble expression level of dAb, with the specific AA combinations being (S, R, N, D, Q), (G, R, C, N, S) and (R, S, G); these combinations respectively affected the dAb expression level in the broth supernatant, the level in the pellet lysate and total soluble dAb. Among the 20 AAs, R displayed a negative influence on the soluble expression level, whereas G and S showed positive effects. A linear model was built to predict the soluble expression level from the sequence; this model had a prediction accuracy of 80 %. In summary, increasing the content of polar AAs, especially G and S, and decreasing the content of R, was helpful to improve the soluble expression level of HSA dAb.

SUMMARY Lactobacillus plantarum O1 was isolated from the gut of sea bream (Sparus aurata) and identified with the API biochemical test and MALDI-TOF MS. This strain was further characterised according to the selection criteria for lactic acid bacteria as starter cultures for aquatic food production. L. plantarum O1 showed good antimicrobial activity against pathogenic test microorganisms. Further investigation confirmed it as the producer of the bacteriocin plantaricin. This strain also showed good growth at a wide range of temperatures (from 4 to 45 °C) and a wide range of pH (2–12), even in the presence of 3.5% NaCl. Its viability was also good after lyophilisation and in simulated gastric and small intestinal juice. The strain is a promising probiotic, and our further research will focus on its application in the biopreservation of fresh fish and shellfish.

γ-Decalactone is a flavour compound that when obtained by biotechnological production using microorganisms is classified as natural. The aim of this study is to evaluate various conditions for γ-decalactone production by tropical yeast strains Yarrowia lipolytica CCMA 0242 and Lindnera saturnus CCMA 0243. The growth of and γ-decalactone production by Y. lipolytica CCMA 0242 were higher in castor oil than in glycerol. γ-Decalactone production in single batch or fed-batch fermentation did not differ significantly. The γ-decalactone production by L. saturnus CCMA 0243 was better at initial pH=5, while the production by Y. lipolytica CCMA 0242 was better at initial pH=6. The yeast L. saturnus CCMA 0243 produced more γ-decalactone than Y. lipolytica CCMA 0242 under the same fermentation conditions. The crude glycerol was not an alternative substrate for γ-decalactone production by Y. lipolytica CCMA 0242. Castor oil at volume fraction of 30 % showed better results as a substrate. The strain L. saturnus CCMA 0243 showed better results of γ-decalactone production. This yeast species can be considered an alternative producer of γ-decalactone in biotechnological processes.

The present study was done to optimize the power ultrasound processing for maximizing diastase activity of and minimizing hydroxymethylfurfural (HMF) content in honey using response surface methodology. Experimental design with treatment time (1-15 min), amplitude (20-100 %) and volume (40-80 mL) as independent variables under controlled temperature conditions was studied and it was concluded that treatment time of 8 min, amplitude of 60 % and volume of 60 mL give optimal diastase activity and HMF content, i.e. 32.07 Schade units and 30.14 mg/kg, respectively. Further thermal profile analyses were done with initial heating temperatures of 65, 75, 85 and 95 ºC until temperature of honey reached up to 65 ºC followed by holding time of 25 min at 65 ºC, and the results were compared with thermal profile of honey treated with optimized power ultrasound. The quality characteristics like moisture, pH, diastase activity, HMF content, colour parameters and total colour difference were least affected by optimized power ultrasound treatment. Microbiological analysis also showed lower counts of aerobic mesophilic bacteria and in ultrasonically treated honey than in thermally processed honey samples complete destruction of coliforms, yeasts and moulds. Thus, it was concluded that power ultrasound under suggested operating conditions is an alternative nonthermal processing technique for honey.

The most severe effects of rheumatoid arthritis (RA) are loss of physical function and chronic pain, which may have a major impact on different areas of the person’s existence. The aim of this study was to get an insight into the quality of life (QOL) in subjects with RA in connection with pain perception and functional ability. The following instruments were used: the World Health Organization Quality of Life Questionnaire (WHOQOL-BREF), Short Form Health Survey (SF-36), Health Assessment Questionnaire Disability Index (HAQ-DI) and Visual Analog Scale for Pain (VAS Pain). The results indicated that there was no statistical difference in the QOL between subjects with RA and healthy population according to SF-36 Croatian norms. Also, the results showed that stronger pain experience was significantly associated with poorer social functioning assessment (SF36SF, Spearman’s rho=-0.463, p, Neke od najozbiljnijih posljedica reumatoidnog artritisa (RA) su kronična bol i gubitak fizičkih funkcija, što može imati snažan utjecaj na različita područja osobne egzistencije u oboljelih. U tom smislu, cilj ovoga istraživanja odnosio se na dobivanje uvida u kvalitetu života osoba s RA u povezanosti s percepcijom boli i funkcionalnom sposobnošću. Korišteni su sljedeći instrumenti procjene: Upitnik kvaliteta života Svjetske zdravstvene organizacije (WHOQOL-BREF), Upitnik za samoprocjenu zdravstvenog stanja SF-36, Upitnik procjene zdravstvenog statusa i indeksa invalidnosti (HAQ-DI) i Vizualno analogna ljestvica samoprocjene boli (VAS). Prema dobivenim podacima nije utvrđena statistički značajna razlika u procjeni kvalitete života između osoba s RA i zdrave populacije prema standardiziranoj hrvatskoj verziji upitnika SF-36. Međutim, intenzivniji doživljaj boli bio je značajno povezan s lošijom procjenom socijalnog funkcioniranja (SF36SF, Spearmanov rho=-0,463, p

The production of γ-linolenic acid (GLA) by the fungus Cunninghamella elegans CFR C07 in submerged fermentation was studied. Culture parameters such as carbon source and incubation time were optimized. Four different extraction methods using solvents with acid washed sand, glass beads, lyophilization and Soxhlet extraction were evaluated for improved extraction of lipids from the fungal biomass after fermentation. The GLA production was initially optimized in 250-mL flask and the process was demonstrated in a 3-litre fermentor. The maximum GLA production was 882 mg/L in shake flask culture and 733 mg/L in the fermentor. The study shows that Cunninghamella elegans CFR C07 is a potent organism for the production of GLA under submerged conditions.

Sage (Salvia sp.) is a genus of a native Mediterranean plant used for aromatization of foods. In this study, total polyphenolic profile, antioxidant properties and antiplatelet effect of short-toothed sage (Salvia brachyodon Vandas) are analyzed and compared to most commonly used Salvia species (S. officinalis, Dalmatian sage). Content of total flavonoids was 0.08-0.23 % and of phenolic acids 0.47-3.04 %. Antioxidant DPPH assay showed higher antioxidant capacity of S. brachyodon (29-36 mg/mL of gallic acid equivalents) than of S. officinalis. In functional test of primary haemostasis, extracts of S. brachyodon have prevented platelet aggregation in nanomolar concentration (21 nM), thus showing potential in prevention of thrombus formation as functional food or dietary supplement. Antiplatelet activity was related to antioxidant capacity (r=0.7014, p=0.0352) indicating that prevention of aggregation is not caused by an individual component, but it is rather a result of synergistic effect of polyphenols. The obtained results are preliminary and a more detailed study of the possibility of applying the investigated plant species as supplements for the prevention of the problem of cardiovascular system and platelet aggregation is needed.

Seven isolates from spoiled fruits and vegetables were screened for pectinase produc¬tion using pectin agar plates and the most efficient bacterial strain, MPTD1, was identified as Bacillus sonorensis. Optimisation of various process parameters was done using Plack¬ett-Burman and Box-Behnken designs and it was found that parameters like yeast extract, K2HPO4, incubation time, NaNO3 and KCl have a negative impact on pectinase production. Parameters like pH and MgSO4 and pectin mass fractions have a positive impact on pecti¬nase production. The maximum obtained enzyme activity was 2.43 (μM/mL)/min. This is the first report on pectinase production by Bacillus sonorensis.

Jumbo squid (Dosidicus gigas) muscle proteins show low functionality with limited use in gel products. This work aims to assess the influence of adding the natural and commercially available fibre, amidated low-methoxyl pectin (at 0.5, 1.0, 1.5, 2.0 and 3.0%), on the physicochemical and functional characteristics of jumbo squid (Dosidicus gigas) mantle muscle gels. The addition of 0.5% fibre showed an immediate effect on the gel texture profile analysis, improving hardness (p

Eggplant (Solanum melongena L.) is one of the most consumed vegetables in the world. The eggplant glycoalkaloids (GAs) are toxic secondary metabolites that may have detrimental effects on human health, particularly if the magnitudes of GAs are higher than the recommended food safety level (200 mg per kg of fresh mass). In this study, the content of solasonine compound and the expression patterns of solasodine galactosyltransferase (SGT1) gene were assessed in different tissues (mature leaves, flower buds, young, mature, and physiologically ripe fruits) of two Iranian eggplant genotypes (D1 and J10) under field conditions. The maximum mass fraction of solasonine in D1 was detected in flower buds (135.63 µg/g), followed by leaf (113.29 µg/g), physiologically ripe fruit (74.74 µg/g), young fruit (61.33 µg/g), and mature fruit (21.55 µg/g). Comparing both genotypes, the genotype of bitter fruits (J10) contained higher mass fraction of solasonine, as one of the main factors for producing bitter flavour of the plant. Regarding the expression profiles of SGT1, in both genotypes, the activity of the gene was increased nearly parallel with the concentration of solasonine. In the J10 genotype, transcript level of the gene was significantly higher than the genotype of sweet fruits (D1). Although both D1 and J10 genotypes are possibly recommendable for human food consumption, D1 is more suitable for daily diet.

Za dobivanje zdravih proizvoda u industrijskoj proizvodnji mesnih emulzija važno je smanjiti udjel fosfata. Ispitan je utjecaj sedam vrsta fosfata na fizikalno-kemijska svojstva te kakvoću proizvoda radi odabira najoptimalnijeg spoja za proizvodnju kuhanih kobasica. Zatim je udjel fosfata postupno smanjivan. Dodatkom tetranatrijevog difosfata i natrijevog tripolifosfata povećala se pH-vrijednost, umanjile strukturne značajke te postigla najbolja stabilnost emulzije i najmanji kalo kuhanja, bez značajnog utjecaja na tvrdoću. Na osnovi viskoelastičnih svojstava zaključeno je da je za dobivanje proizvoda prihvatljive kakvoće dovoljan dodatak minimalnog masenog udjela tetranatrijevog difosfata od 0,06 %. Reološka su se ispitivanja pokazala prikladnima za ocjenu kakvoće mesnih proizvoda, jer su pružila uvid u strukturne značajke proizvoda, a osobito funkcionalna svojstva proteina iz mesa. Na osnovi dobivenih rezultata zaključeno je da se udjel fosfata u mesnim emulzijama može bitno smanjiti, bez negativnih utjecaja na kakvoću proizvoda., Phosphate reduction is of important industrial relevance in the manufacturing of emulsified meat products because it may give rise to a healthier product. The effect of seven different phosphate types was tested on the physicochemical and quality characteristics to select the most promising phosphate type for further cooked sausage manufacturing. Next, phosphate mass fraction was gradually reduced. Tetrasodium di- or pyrophosphate (TSPP) and sodium tripolyphosphate (STPP) increased pH, reduced structural properties, resulted in the highest emulsion stability, lowest cooking loss and had little effect on hardness. Based on the viscoelastic properties, a minimum mass fraction of 0.06 % TSPP was sufficient to obtain an acceptable quality product. Rheology proved to be a very useful tool to evaluate the quality of meat products, as it gives insight in the structure of the meat product and especially the functional properties of meat proteins. Based on the obtained results, it can be concluded that the current amount of phosphate added to emulsified meat products can be significantly reduced with minimal loss of product quality.

This study evaluated the use of three solid brewery wastes: brewer’s spent grain, hot trub and residual brewer’s yeast, as alternative media for the cultivation of lactic acid bacteria to evaluate their potential for proteolytic enzyme production. Initially, a mixture experimental design was used to evaluate the effect of each residue, as well as different mixtures (with the protein content set at 4 %) in the enzyme production. At predetermined intervals, the solid and liquid fractions were separated and the extracellular proteolytic activity was determined. After selecting the best experimental conditions, a second experiment, factorial experimental design, was developed in order to evaluate the protein content in the media (1 to 7 %) and the addition of fermentable sugar (glucose, 1 to 7 %). Among the wastes, residual yeast showed the highest potential for the production of extracellular enzymes, generating a proteolytic extract with 2.6 U/mL in 3 h. However, due to the low content of the fermentable sugars in the medium, the addition of glucose also had a positive effect, increasing the proteolytic activity to 4.9 U/mL. The best experimental conditions of each experimental design were reproduced for comparison, and the enzyme content was separated by ethanol precipitation. The best medium produced a precipitated protein with proteolytic activity of 145.5 U/g.

In the current study, the antibacterial effect of Echinophora platyloba essential oil and common liquid smoke (individually and in combination) against Staphylococcus aureus in beef meat samples is investigated. Using an automated microbiological growth analyser and the turbidimetric technique, the minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) of the essential oil and liquid smoke were determined. Anti-S. aureus activity of essential oil and liquid smoke (individually and in combination) was defined by disk diffusion assay, generation time and cell constituent release. Apart from that, the interactions between these two compounds were measured by the checkerboard assay and by calculating the fractional inhibitory concentration (FIC) indices. Related MIC values of essential oil and smoke were found to be 7200 and 5500 mg/L, and MBC values were 8500 and 8000 mg/L, respectively. The conducted organoleptic assay showed that the addition of 0.05 g of essential oil and 0.6 g of liquid smoke to 100 g of meat samples did not have adverse effect on the overall acceptance. Weaker antibacterial effect against Staphylococcus aureus was observed when only Echinophora platyloba essential oil was used than when it was used in combination with liquid smoke.

The effect of surfactants on the bioreduction of acetophenones mediated by carrot (Daucus carota) root was investigated. Among the tested surfactants, Tween® 20 slightly improved the conversion. The amount of surfactant was optimised, and the presented methodology was applied to other substituted acetophenones, which led to the preparation of their respective optically enriched 1-phenylethanols with high conversion levels and high enantioselectivities.

The study examines the potential of wine industry by-product, the lees, as a rich mixture of natural polyphenols, and its physiological potential to reduce postprandial metabolic and oxidative stress caused by a cholesterol-rich diet in in vivo model. Chemical analysis of wine lees showed that their total solid content was 94.2%. Wine lees contained total phenols, total nonflavonoids and total flavonoids expressed in mg of gallic acid equivalents per 100 g of dry mass: 2316.6±37.9, 1332.5±51.1 and 984.1±28.2, respectively. The content of total anthocyanins expressed in mg of cyanidin-3-glucoside equivalents per 100 g of dry mass was 383.1±21.6. Antioxidant capacity of wine lees determined by the DPPH and FRAP methods and expressed in mM of Trolox equivalents per 100 g was 259.8±1.8 and 45.7±1.05, respectively. The experiment lasted 60 days using C57BL/6 mice divided in four groups: group 1 was fed normal diet and used as control, group 2 was fed normal diet with added wine lees, group 3 was fed high-cholesterol diet (HCD), i.e. normal diet with the addition of sunflower oil, and group 4 was fed HCD with wine lees. HCD increased serum total cholesterol (TC) by 2.3-fold, triacylglycerol (TAG) by 1.5-fold, low-density lipoprotein (LDL) by 3.5-fold and liver malondialdehyde (MDA) by 50%, and reduced liver superoxide dismutase (SOD) by 50%, catalase (CAT) by 30% and glutathione (GSH) by 17.5% compared to control. Conversely, treatment with HCD and wine lees reduced TC and LDL up to 1.4 times more than with HCD only, with depletion of lipid peroxidation (MDA) and restoration of SOD and CAT activities in liver, approximating values of the control. HDL levels were unaffected in any group. Serum transaminase activity showed no hepatotoxic properties in the treatment with lees alone. In the proposed model, wine lees as a rich polyphenol source could be a basis for functional food products without alcohol.

Nodule bacteria (rhizobia) in symbiotic associations with legumes enable considerable entries of biologically fixed nitrogen into soil. Efforts are therefore made to intensify the natural process of symbiotic nitrogen fixation by legume inoculation. Studies of field populations of rhizobia open up the possibility to preserve and probably exploit some indigenous strains with hidden symbiotic or ecological potentials. The main aim of the present study is to determine genetic diversity of common bean rhizobia isolated from different field sites in central Croatia and to evaluate their symbiotic efficiency and compatibility with host plants. The isolation procedure revealed that most soil samples contained no indigenous common bean rhizobia. The results indicate that the cropping history had a significant impact on the presence of indigenous strains. Although all isolates were found to belong to species Rhizobium leguminosarum, significant genetic diversity at the strain level was determined. Application of both random amplification of polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC- -PCR) methods resulted in similar grouping of strains. Symbiotic efficiency of indigenous rhizobia as well as their compatibility with two commonly grown bean varieties were tested in field experiments. Application of indigenous rhizobial strains as inoculants resulted in significantly different values of nodulation, seed yield as well as plant nitrogen and seed protein contents. The most abundant nodulation and the highest plant nitrogen and protein contents were determined in plants inoculated with R. leguminosarum strains S17/2 and S21/6. Although, in general, the inoculation had a positive impact on seed yield, differences depending on the applied strain were not determined. The overall results show the high degree of symbiotic efficiency of the specific indigenous strain S21/6. These results indicate different symbiotic potential of indigenous strains and confirmed the importance of rhizobial strain selection. These are the first studies of indigenous common bean rhizobia in Croatia that provide the basis for further characterization and selection of highly efficient indigenous strains and their potential use in agricultural practice and future research.

To differentiate monovarietal wines made from native and introduced varieties in Istria (Croatia), samples of Malvazija istarska, Chardonnay and Muscat yellow from two harvest years (2013 and 2014) were subjected to headspace solid-phase microextraction and gas chromatographic/mass spectrometric analysis (HS-SPME-GC/MS) of volatile aroma compounds. Significant effects of variety and harvest year were determined, but their interaction complicated the differentiation. Particular compounds were consistent as markers of variety in both years: nerol for Malvazija, ethyl cinnamate and a tentatively identified isomer of dimethylbenzaldehyde for Chardonnay, and terpenes for Muscat yellow. Wines from 2013 contained higher concentrations of the majority of important volatiles. A 100% correct differentiation of Malvazija istarska and Chardonnay wines according to both variety and harvest year was achieved by stepwise linear discriminant analysis.

Uvod: Sindrom pekućih usta (SPU) kronični je poremećaj koji uglavnom zahvaća žene u postmenopauzi, a svojstven mu je osjećaj pečenja u usnoj šupljini unatoč potpuno zdravoj oralnoj sluznici. Rezultati dosadašnjih istraživanja sugeriraju moguću ulogu centralnih i/ili perifernih neuroloških poremećaja. Ispitanici i postupci: U skupini od 20 ispitanika sa SPU-om (64,7 ± 12,3 godina) i njih 20 u kontrolnoj skupini (61,5 ± 15 godina) s bolovima u lumbosakralnom dijelu leđa, transkranijalnom sonografijom evaluirani su moždani parenhim, crna tvar (substantia nigra), središnji niz stanica smješten u formatio reticularis lateralno od rapha medulla oblongata (nucleus raphe) i crvena jezgra (nucleus ruber). Statistička analiza obavljena je Studentovim t-testom sa značajnošću od p < 0,05. Rezultati: Rezultati ovog istraživanja pokazuju da se kod oboljelih od SPU-a (p < 0,05), u usporedbi s kontrolnom skupinom, pojavljuje hipoehogenost substantia nigra i nuclei raphe, a hiperehogen ostaje nucleus ruber. Zaključak: Možemo zaključiti da promijenjeni nalazi transkranijalne sonografije moždanoga parenhima, nucleus raphe i crvene jezgre (nucleus ruber) kod bolesnika sa SPU-om mogu odražavati centralne poremećaje u sklopu ovog sindroma., Objective: Burning mouth syndrome (BMS) is a chronic disorder which is affecting mostly postmenopausal women and is characterized by burning symptoms in the oral cavity on the clinically healthy oral mucosa. The results of previous studies suggested a possible role of peripheral and/or central neurological disturbances in these patients. The aim of this study was to analyze patients with burning mouth syndrome using transcranial sonography. Methods: By use of transcranial sonography of the brain parenchyma, substantia nigra, midbrain raphe and brain nucleus were evaluated in 20 patients with BMS (64.7±12.3 years) and 20 controls with chronic pain in the lumbosacral region (61.5±15). Statistical analysis was performed by use of Student t test with significance set at p

Coffee bean fermentation is a spontaneous, on-farm process involving the action of different microbial groups, including bacteria and fungi. In this study, high-throughput sequencing approach was employed to study the diversity and dynamics of bacteria associated with Brazilian coffee bean fermentation. The total DNA from fermenting coffee samples was extracted at different time points, and the 16S rRNA gene with segments around the V4 variable region was sequenced by Illumina high-throughput platform. Using this approach, the presence of over eighty bacterial genera was determined, many of which have been detected for the first time during coffee bean fermentation, including Fructobacillus, Pseudonocardia, Pedobacter, Sphingomonas and Hymenobacter. The presence of Fructobacillus suggests an influence of these bacteria on fructose metabolism during coffee fermentation. Temporal analysis showed a strong dominance of lactic acid bacteria with over 97% of read sequences at the end of fermentation, mainly represented by the Leuconostoc and Lactococcus. Metabolism of lactic acid bacteria was associated with the high formation of lactic acid during fermentation, as determined by HPLC analysis. The results reported in this study confirm the underestimation of bacterial diversity associated with coffee fermentation. New microbial groups reported in this study may be explored as functional starter cultures for on-farm coffee processing.

Enzimska interesterifikacija koristi se za obradu ulja i masti radi dobivanja restrukturiranih lipida sa željenim tehnološkim svojstvima. Međutim, s obzirom na to da sirovi materijali sadržavaju velike udjele bioaktivnih tvari, još uvijek je nejasan utjecaj ovakvih enzimskih procesa na biološku aktivnost konačnog produkta. Stoga je svrha ovog istraživanja bila procijeniti antioksidacijski potencijal i modulacijski učinak dvaju sirovih materijala iz područja Amazone (buriti ulja i murumuru masti) na stanice jetre (HepG2), i to prije i nakon procesa interesterifikacije. Istraživanje je pokazalo da su neke bioaktivne komponente sirovih materijala i njihov antioksidacijski učinak očuvani nakon enzimske esterifikacije restrukturiranih lipida, pri čemu dolazi do modulacije HepG2 endogenog antioksidacijskog enzima., Enzymatic interesterification is used to manipulate oil and fat in order to obtain improved restructured lipids with desired technological properties. However, with raw materials containing significant amounts of bioactive compounds, the influence of this enzymatic process on the bioactivity of the final product is still not clear. Thus, the aim of this study is to evaluate the antioxidant potential and modulatory effects of two raw materials from the Amazonian area, buriti oil and murumuru fat, before and after lipase interesterification, on human hepatoma cells (HepG2). The results indicate that minor bioactive compounds naturally found in the raw materials and their antioxidant capacity are preserved after enzymatic interesterification, and that the restructured lipids modulate HepG2 endogenous antioxidant enzyme.

This research was undertaken to investigate the infl uence of extraction parameters during microwave-assisted extraction on total phenolic content, total fl avonoids, total hydroxycinnamic acids and total fl avonols of blackthorn fl owers as well as to evaluate the antioxidant capacity by two diff erent methods (2, 2-diphenyl-1-picrylhydrazyl free radical scavenging capacity and ferric reducing antioxidant power assays). The investigated extraction parameters were: solvent type and volume fraction of alcohol in solvent (50 and 70 % aqueous solutions of ethanol and methanol), extraction time (5, 15 and 25 min) and extraction temperature (40, 50 and 60 °C) controlled by microwave power of 100, 200 and 300 W. Multivariate analysis of variance (MANOVA) was used to evaluate the diff erences at a 95 % confi dence level (p≤0.05). The obtained results show that aqueous solution of ethanol was more appropriate solvent for extraction of phenolic compounds (total fl avonoids, total hydroxycinnamic acids and total fl avonols) than aqueous solution of methanol. The amount of phenolic compounds was higher in 70 % aqueous solution of ethanol or methanol, while higher antioxidant capacity was observed in 50 % aqueous solution of methanol. Higher temperature of extraction improved the amount of phenolic compounds and also antioxidant capacity determined by 2, 2-diphenyl-1-picrylhydrazyl free radical scavenging capacity assay. Extensive duration of extraction (15- to 25-minute interval) has a signifi cant eff ect only on the increase of total phenolic content, while specifi c phenolic compound content and antioxidant capacity were the highest when microwave extraction time of 5 min was applied.

Acidic liquids such as verjuice, lemon juice and vinegar are frequently consumed in Iran. Different kinds of acidic liquids are packaged in polyethylene terephthalate (PET) and high-density polyethylene (HDPE) bottles. There is evidence indicating that phthalates can leach from PET and HDPE bottles into their contents. In this work the effect of storage time, temperature and bottle type on the migration of phthalates from packaging materials into acidic liquids is studied by analyzing the samples stored under different conditions, before storage and after 2, 4 and 6 months of storage. The determined mean phthalate concentrations in µg/L were

The eff ect of 24-hour fermentation of lupin seeds by different yeast strains on their chemical composition was determined. After fermentation, the mass fraction of proteins increased and their in vitro digestibility and biological activity significantly improved. The amino acid profi le of fermented products was similar to that of raw lupin seeds. The significant reduction in the mass fraction of oligosaccharides and phytate, but not of alkaloids was found. The pH level of fermented products decreased as a consequence of the increase of lactic and propionic acid mass fractions. The most favourable changes in the Chemical composition of blue lupin seeds were obtained in fermentation with Saccharomyces cerevisiae baker’s yeast and Fermivin 7013 strain.

Physicochemical properties, consumer acceptance, antioxidant and angiotensin-converting enzyme (ACE) inhibitory activities of infusions and fermented beverages of Eucalyptus camaldulensis and Litsea glaucescens were compared. Among physicochemical parameters, only the pH of fermented beverages decreased compared with the unfermented infusions. No relevant changes were reported in consumer preference between infusions and fermented beverages. Phenolic profile measured by UPLC MS/MS analysis demonstrated significant concentration changes of these compounds in plant infusions and fermented beverages. Fermentation induced a decrease in the concentration required to stabilize 50% of DPPH radical (i.e. lower IC50). Additionally, it enhanced the antioxidant activity measured by the nitric oxide scavenging assay (14% of E. camaldulensis and 49% of L. glaucescens); whereas relevant improvements in the fermented beverage were not observed in the lipid oxidation assay compared with unfermented infusions. The same behaviour was observed in the inhibitory activity of ACE; however, both infusions and fermented beverages had lower IC50 than positive control (captopril). The present study demonstrated that fermentation has an influence on the concentration of phenolics and their potential bioactivity. E. camaldulensis and L. glaucescens can be considered as natural sources of biocompounds with antihypertensive potential used either as infusions or fermented beverages.

In order to detect thermotolerant Campylobacter spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase chain reaction/restriction enzyme analysis method for exact species determination. Campylobacter spp. prevalence was 73.86 %. Campylobacter jejuni and Campylobacter coli were isolated from 53.53 and 15.35 % of the samples, respectively. In 4.98 % of isolates final species of thermotolerant Campylobacter spp were not determined. The multilocus sequence typing method was used to evaluate genetic diversity of eight Campylobacter jejuni and four Campylobacter coli isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs) and clonal complexes (CCs) of Campylobacter jejuni and C. coli isolates in Croatia. By applying the multilocus sequence typing, a new allele of tkt gene locus was discovered and marked as tkt508. The C. jejuni ST 6182 and C. coli ST 6183 genotypes were described for the first time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of Campylobacter jejuni and Campylobacter coli in Croatia.

Fruits and vegetables are considered a good source of polyphenols and antioxidant capacities which are beneficial in protecting the human body against damage induced by reactive species. The objective of this work is to conduct an assessment of the polyphenol content and antioxidant activities of diff erent fruit (kiwi, pear, green apple, raspberry, blackberry, strawberry and blueberry) and vegetable (pumpkin, green and red pepper) extracts using both chemical extraction and a modified in vitro digestive enzymatic extraction in order to compare results. Polyphenol content and antioxidant capacity of different fruits, vegetables and fruit juices were determined by Folin-Ciocalteu and FRAP methods, respectively. It was observed that polyphenol content expressed as gallic acid equivalents of extracts obtained with the two extraction methods was signifi cantly (p

Corynebacterium parvum used alone to enhance immunological reactivity produced transient inhibition of the growth of chemically induced isogenic mouse tumours. Attempts were made to combine C. Parvum with cyclophosphamide to see whether this would increase the latter's effectiveness in inhibiting early but established tumours. Of the various regimens tested, the administration of the C. parvum 12 days after a single dose of chemotherapy produced dramatic inhibition of tumour growth and resulted in complete and lasting regressions in up to 70% of tumour-bearing animals. The most important variable in this regimen is the time between the chemotherapy and the subsequent immunotherapy.It is possible that non-specific active immunotherapy with agents such as C. parvum may be a valuable adjunct to the conventional cyto-reductive treatments of cancer, but the time of administration of such therapy is probably critical for each tumour and for each chemotherapeutic regimen.

The pattern of free amino-acid concentrations in maternal venous plasma, fetal umbilical arterial plasma, fetal urine, and amniotic fluid at 15 to 20 weeks9 gestation has been determined. Free amino-acid concentrations were greater in fetal plasma than in maternal plasma, amniotic fluid, or fetal urine. The ratios of amino-acid concentrations in fetal umbilical arterial plasma and urine indicate that the fetal kidney can effectively conserve amino-acids, possibly reaching an adult level of competence in this respect. There was little correlation between amino-acid concentrations in the fluids analysed with the exception of that between amniotic fluid and fetal urine.

Rifampicin showed high activity against experimental leprosy, inhibiting the multiplication of dapsone-sensitive and dapsone-resistant strains of Mycobacterium leprae in mice fed 5 mg./kg. body weight. In a formal pilot-type trial on six previously untreated patients with active lepromatous leprosy, rifampicin (600 mg. daily by mouth) was as effective as standard treatment with dapsone. Myco. leprae, however, appeared to be killed more rapidly by rifampicin than by dapsone or other antileprosy drugs so far studied. This was confirmed on a further 10 patients, including two with dapsone resistance, and from the infectivity in mice of bacilli recovered from patients during treatment with rifampicin or dapsone. These results are consistent with the bactericidal activity of rifampicin against other micro-organisms, which could be important to the chemotherapy of leprosy, since all antileprosy drugs in current use are bacteriostatic.

The leucocyte migration test, an in-vitro measure of cellular immunity, has been used to follow the changes in cell-mediated hypersensitivity to kidney and histocompatibility antigens in three patients after renal transplantation. Inhibition of leucocyte migration, indicating strong sensitization to the antigens used, occurred in each patient, starting five to seven days after transplantation. Satisfactory renal function had not been established in any of the patients at this time. In one case inhibition of leucocyte migration persisted almost continuously until the 24th day and was associated with poor renal function proved histologically to be due to rejection. Treatment with increased dosage of prednisone was associated with a rapid reversion to normal of the migration index and improvement in renal function. Later, inhibition of migration occurred again, and shortly afterwards the graft ceased to function. In the other two cases the migration index became normal without alteration in immunosuppressive therapy and a satisfactory diuresis followed. It is suggested that this simple test should prove useful in the specific diagnosis of rejection and in control of immunosuppressive therapy.