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Background: Our study compared the haemodynamic changes after spinal anesthesia with 2% lignocaine and 0.5% plain bupivacaine.Methods: A controlled, randomized trial was performed on 30 patients scheduled for arthroscopic knee surgery. 2% lignocaine and 0.5% plain bupivacaine were used for spinal anaesthesia. We measured cardiac output (electrical bioimpedance cardiography), blood pressure and development of sensory blockade before and for 25 minutes after spinal anaesthesia.Results: In patients developing a sensory block below T6 there were no differences between both anaesthetics in haemodynamic parameters. But in patients developing a sensory block at or above T6 there was a greater drop in mean arterial pressure and cardiac output and a faster decrease in heart rate for bupivacaine compared to patients receiving Lignocaine.Conclusion: In patients developing a sensory block at or above the T6 dermatome, the decrease in cardiac output and mean arterial pressure in the first 25 min after spinal anaesthesia is smaller when 2% lignocaine rather than 0.5% bupivacaine is used for blockade. [ABSTRACT FROM AUTHOR]

Background/Aim. Intracranial aneurysms are pathological enlargement of the wall of cerebral arteries. Intracranial aneurysms rupture is a dramatic event with a significant morbidity and mortality. The Fisher Grade is widely accepted in assessment of the extensiveness of aneurysmal subarachnoid hemorrhage (aSAH) and the presence of other intracranial hemorrhage on the computed tomography (CT) scan. Significant early complication of a aSAH may be a cerebral vasospasm. The aim of this study was to determine the relationship between the extensiveness of aSAH, assessed by the Fisher Grade on admission, with the intensity of cerebral vasospasm in patients with ruptured intracranial aneurysm. Methods. This prospective clinical study included 50 patients with aSAH hospitalized at the Clinic of Neurosurgery, Clinical Center of Vojvodina, Novi Sad, Serbia. All the patients underwent 256-layer cranial CT and CT angiography on admission and on the day 9. Based on native CT scans, they were classified according to the Fisher Grade. On CT angiography images, intensity of cerebral vasospasm was determined. Results. On the basis of admission CT images, 24% of patients were classified into the Fisher Grade group 2, while 34% and 42% were in the groups 3 and 4, respectively. A positive correlation of the Fischer Grade on admission with the intensity of cerebral vasospasm was established, but with no statistical significance (ρ = 0.273, p = 0.160). Conclusion. This study showed that the Fisher Grade is not significant in predicting the intensity of cerebral vasospasm in patients hospitalized with intracranial aneurysm rupture.

Background: Fraud attempts create large losses for financing subjects in modern economies. At the same time, leasing agreements have become more and more popular as a means of financing objects such as machinery and vehicles, but are more vulnerable to fraud attempts. Objectives: The goal of the paper is to estimate the usability of the data mining approach in discovering fraud in leasing agreements. Methods/Approach: Real-world data from one Croatian leasing firm was used for creating tow models for fraud detection in leasing. The decision tree method was used for creating a classification model, and the CHAID algorithm was deployed. Results: The decision tree model has indicated that the object of the leasing agreement had the strongest impact on the probability of fraud. Conclusions: In order to enhance the probability of the developed model, it would be necessary to develop software that would enable automated, quick and transparent retrieval of data from the system, processing according to the rules and displaying the results in multiple categories.

Background/Aim. At the moment there are few scoring systems for malignant astrocytoma but they are not widely accepted. The aim of this study was to evaluate malignant astrocytoma score (MAS) on a new group of patients with malignant astrocytoma, to compare MAS with other prognostic tools and to describe the use of MAS in everyday practice in neurooncology. Methods. The study was performed on 124 patients with supratentorial malignant astrocytoma grade III or IV. They were operated on and subsequently irradiated with 50-60 Gy. Results. The mean age of the patients was 57.3 years. The mean Karnofski performance status (KPS) of the functional inpairment was 54. The removal of the tumor > 90% was done in 59.7% of patients. The mean survival was 9.1 months, and 27.4% of patients had a 12-month survival. The area under receiver operating characteristic (ROC) curve (AUC) of the MAS for predicting 6-, 12- and 18-month survival was 0.754, 0.783 and 0.882, respectively. We compared the MAS with the two mostly cited scoring systems. The AUC for the same prediction for medical research council (MRC) was 0.601, 0.693, 0.772 respectively. For the Radiation Therapy Oncology Group (RTOG) the AUC was 0.732, 0.765, 0.827, respectively. Conclusion. MAS represents a useful scoring system for determining illness severity and prognosis in patients with malignant supratentorial astrocytoma. It can be helpful in comparing single patients or groups of patients, as well as results of different treatments and in controlling the quality of hospital treatment and so on.

Introduction. A gunshot head injury, characterized by a huge intensity of mechanical force, in addition to the direct tissue damage at the location of direct impact, may cause a skull and skull base fracture, distant from the the point of direct impact, which could be further complicated by creating a communication between endocranium and nasal/paranasal cavities. Such cases pose a great diagnostic and therapeutic challenge for every clinician. Case report. The patient is presented with the history of a perforating gunshot head injury six years ago, with recurrent attacks of meningoencephalitis subsequently, without rhinorrhea. By using high resolution CT scans, previous traumatic skull injury was verified and a fissure in the frontoethmoidal region, far from the point of direct impact, was detected. The patient underwent transnasal endoscopic surgery, in order to seal the communication on skull basis. The patient did not suffer from meningoencephalitis during the next two years. Conclusion. In the cases with late occurrence of posttraumatic meningoencephalitis with no signs of rhinorrhea, a possibility of an existing communication between intracranial and nasal cavities should be considered, as well. By applying modern diagnostic and therapeutic procedures such communication should be precisely located and sealed.

Ion exchange chromatography and SDS-PAGE followed by identification by MALDI-TOF/TOF, and two-dimensional gas chromatography with time-of-flight mass spectrometry (GC × GC-TOF-MS) were used for comprehensive proteomics and volatilomics evaluation of the effects of pre-fermentative oxygenation, skin contact and use of pectolytic enzymes in production of Malvazija istarska white wine, respectively. Many protein species and an unprecedented number of volatiles have been identified and (semi)quantified, revealing high complexity of the observed effects. Compared to a standard control wine, oxygenation treatment modulated the protein composition and resulted with a volatilome characterized by decreased levels of several important volatiles. Skin contact treatments, especially in combination with pectolytic enzymes, significantly increased the levels of a large number of proteins, but were also deprived of particular protein species found in other wines. Wines obtained by skin contact with exogenous enzymes exhibited the most complex volatile composition with increased levels of many key monoterpenoids, alcohols and esters., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Introduction: Oral cancer (OC) is a disease with a high mortality rate due to its late recognition. Since the oral cavity is easily accessible for visual inspection, enabling early diagnosis, the education of healthcare workers about preventive oral examinations is critical. This research aimed to assess the level of participants' OC knowledge, as well as to raise awareness about this diagnosis., Materials and Methods: The research was conducted as an online survey among students of dental medicine, students of medicine, doctors of dental medicine and doctors of medicine. The questionnaire was designed solely for the purpose of this study and consisted of 29 questions. The first part of the questionnaire consisted of general questions about the participants, whereas the questions in the second part addressed their knowledge and attitudes towards OC., Results: The surveyed population comprised of 140 dental students, 105 medical students, 159 doctors of dental medicine and 100 medical doctors. The level of knowledge about OC among the participants is not yet satisfactory. The group of dental medicine students scored highest, while medical doctors showed the weakest knowledge., Conclusion: Additional education about OC for doctors of dental medicine and medical doctors is needed. This step will improve prevention and increase chances for early detection.

Six different vinification treatments, including a control treatment (7-day standard maceration) (K7), were performed to study the effects of non-standard techniques on bioactive compounds and sensory attributes of Teran red wine. Pre-fermentative mash cooling (8 °C; 48 h) and heating (50 °C; 48 h) followed by prolonged post-fermentative maceration of 13 days (C15;H15) or 28 days (C30;H30) were applied. In another treatment, after cooling, saignée was performed followed by 13-day prolonged maceration (CS15). Wine phenols and vitamins were analyzed by HPLC-DAD-FLD, minerals by ICP-OES, and sensory analysis was performed using the QDA and 100-point O.I.V./U.I.O.E. methods. Obtained results showed total phenolic concentration was the highest in the H30 treatment. The concentration of anthocyanins, flavan-3-ols and phenolic acids was significantly higher in wines of all vinification techniques compared to the control. Stilbene content was highly affected by pre-fermentative heating. Treatments CS15, H15, C30 and H30 resulted in the highest scores by both the QDA and 100-point sensory methods. The obtained results suggest that advanced non-standard vinification techniques have a significant impact on Teran wine by enhancing its composition of bioactive compounds and improving its sensory profile, which gives it an additional market value. Furthermore, a comprehensive comparison of such techniques applied simultaneously in one study is of substantial importance for additional research in wine production.

Metabolomics has evolved into a particularly useful tool to study interactions between metabolites and serves as an aid in unraveling the complexity of entire metabolomes. Nonetheless, it is increasingly viewed as a methodology with practical applications in the clinical setting, where identifying and quantifying biomarkers of interest could prove useful for diagnostics. Starting from a concise overview of the most prominent analytical techniques employed in metabolomics, herein we present a review of its application in studies of brain metabolism and cerebrovascular diseases, paying most attention to its uses in researching aneurysmal subarachnoid hemorrhage. Both animal models and human studies are considered, and metabolites identified as potential biomarkers are highlighted., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Lasica, Raicevic, Stojanovic, Djilvesi, Horvat, Jelaca, Pajicic and Vulekovic.)

There is a lack of studies evaluating the metabolic contribution of non- Saccharomyces yeasts in early fermentation phases. This study aimed to investigate the volatile aroma profiles produced by various non- Saccharomyces yeasts just before sequential inoculation with Saccharomyces cerevisiae to provide an insight into the particular effects they induce at this stage. The grape must of Malvazija istarska was inoculated with monocultures of Torulaspora delbrueckii , Metschnikowia pulcherrima , Pichia kluyveri , Lachancea thermotolerans , and Schizosaccharomyces pombe , alongside a S. cerevisiae control. Eighty volatile compounds were quantified via headspace solid-phase microextraction and gas chromatography-mass spectrometry, and the data were statistically elaborated. Volatile profiles of non- Saccharomyces yeasts differed significantly from the S. cerevisiae control. Most treatments caused increases in linalool and β-damascenone, decreases in higher alcohols and fatty acids, and improved synthesis of odoriferous esters. Torulaspora delbrueckii and M. pulcherrima produced compounds not commonly found in S. cerevisiae fermented wines. Multivariate statistical analysis linked the investigated yeasts to specific, particularly abundant compounds. Future studies should explore to what degree these contributions persist after sequential inoculation with S. cerevisiae in diverse grape must matrices.

Shallots are a perennial plant from the Alliaceae family, classified with the common onion under the name of the Allium cepa Aggregatum group. The term shallot is also used for diploid and triploid viviparous onions, known as Allium × proliferum (Moench) Schrad and Allium × cornutum Clementi ex Vis., respectively. In this study, we compared the dry matter, pyruvic acid content, sugar content, flavonoid content, antioxidant capacity and mineral composition of 34 shallot accessions falling into three shallot species ( Allium × cornutum , Allium × proliferum and A. cepa Aggregatum). Shallot accessions belonging to the A.× cornutum and A. × proliferum groups are characterized by high dry matter content (around 25%), of which a little less than 50% is formed of inulin-type sugars, polysaccharides, considered an excellent prebiotic with beneficial effects on human health. On the other hand, accessions belonging to the A. cepa Aggregatum group have lower dry matter content and, as a result, lower pungency (measured as pyruvic acid content), making them more suitable for fresh consumption by a broader range of consumers, but, at the same time, abundant in phenolic compounds, especially quercetin and isorhamnetin glycosides. We also observed a greater biodiversity among accessions within the A. cepa Aggregatum group in all the analyzed physico-chemical parameters compared to the other shallot groups. The investigated shallot accessions have an excellent in vitro antioxidant capacity, as well as excellent nutritional properties.

The present study aimed to assess the association of 16 polymorphisms in genes encoding prothrombotic and cardiovascular risk factors with COVID-19 disease severity: FV G1691A, FV H1299R, FII G20210A, MTHFR C677T, MTHFR A1298, factor XIII V34L, PAI-1 4G/5G, EPCR haplotypes (A1/A2/A3), eNOS -786 T > C, eNOS G894T, LTA C804A, ACE I/D, ITGB3 PIA1/A2, ITGA2B Baka/b, β-Fbg -455 G > A and ApoB R3500Q. The study included 30 patients with severe COVID-19 and 49 non-severe COVID-19 patients. All studied polymorphisms except ITGA2B Baka/b were determined using multilocus genotyping assays CVD StripAssays (ViennaLab Diagnostics), while ITGA2B was genotyped using a real-time PCR method based on TaqMan technology. A higher frequency of carriers of at least one ITGB3 PIA2 allele was found in severe COVID-19 patients (p = 0.009). The distribution of genotypes was significantly different for ß-Fbg -455 G > A (p = 0.042), with only three homozygous AA genotypes found among severe COVID-19 patients. The association with an increased risk for severe COVID-19 was found for ITGB3, with carriers of at least one ITGB3 PIA2 allele having a 3.5-fold greater risk of severe COVID-19 (p = 0.011). Genotype distribution differences were obtained for the combinations of FV H1299R and FXIII V34L (p = 0.026), ITGB3 PIA1/A2 and ITGA2B Baka/b (p = 0.024), and ACE I/D and PAI-1 4G/5G (p = 0.046). ITGB3 polymorphism emerged as an independent risk factor for severe COVID-19 and homozygosity for ß-Fbg -455 G > A mutation could contribute to disease severity. The combined effect of polymorphisms in genes encoding prothrombotic and cardiovascular risk factors could further contribute to disease severity., (© 2022 Wiley Periodicals LLC.)

Moderate wine consumption is often associated with healthy lifestyle habits. The role of wine as a healthy drink is mainly due to its bioactive compounds, which differ according to various viticultural and enological factors. The aim of the present study was to observe the differences in bioactive compounds of white and red autochthonous Croatian wines, differing in terms of the grape variety and production technology. Our further aim was to explore the effect of their moderate consumption (200 mL per day) over the course of six weeks on some aspects of health in sixty-six healthy individuals. Participants were divided into eight groups depending on the wines consumed, while one group formed a non-consuming control group. Medical examination and laboratory tests were performed before the start and at the end of the consumption period. Systolic and diastolic blood pressure, total cholesterol, and LDL concentrations decreased. Additionally, an increase in HDL concentrations, and serotonin and dopamine levels, was observed. ALT, ALP, and GGT levels did not significantly increase in consumer groups, although alcohol concentration was relatively high in all the wines. Such results support the beneficial effects of wine-derived bioactive compounds on some health aspects resulting from moderate white and red wine consumption.

The onset and manifestations of irritable bowel syndrome (IBS) is associated with several factors, and the pathophysiology involves various central and peripheral mechanisms. Most studies indicate that the management of gut microbiota could significantly affect the improvement of subjective disorders in patients with IBS. Numerous clinical trials have assessed the efficacy of probiotics for IBS with controversial conclusions. Several clinical trials have suggested that probiotics can improve global IBS symptoms, while others only improve individual IBS symptoms, such as bloating scores and abdominal pain scores. Only a few clinical trials have found no apparent effect of probiotics on IBS symptoms. Generally, probiotics appear to be safe for patients with IBS. However, the question of which probiotics should be used for certain IBS subtypes remains unresolved. In everyday practice, the dose of the recommended probiotic remains questionable, as well as how long the probiotic should be used in therapy. The use of probiotics in the M subtype and non-classified IBS is particularly problematic, in which combination therapy should be recommended due to the change in symptoms. Therefore, new approaches are needed in the design of clinical studies that should address certain subtypes of IBS., Competing Interests: Conflict-of-interest statement: We declare that there is no conflict of interest., (©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: Associated with epidemics of obesity, nonalcoholic fatty liver disease (NAFLD) is becoming the most prevalent liver disease worldwide. The cornerstone of therapy for NAFLD is lifestyle intervention, mainly focused on weight loss. Significant weight loss results from energy-restricted diets, regardless of macronutrient distribution. An anti-inflammatory diet was related to lower odds of NAFLD among daily alcohol drinkers and individuals with metabolic syndrome. This study aims to evaluate the effect of an energy-reduced anti-inflammatory diet on liver status in younger adults with obesity after a 6-month follow-up., Methods: A two-arm randomized controlled trial surveyed 81 participants' (mean age, 43 years) anthropometric and body composition changes. Metabolic status was determined with glycaemic and lipid status, inflammatory status with hs-CRP, IL-6, and TNF- α , and liver status with liver enzymes, NAFLD-FLS, FLI, and FIB-4 indices. The inflammatory potential of the diet was assessed by the Dietary Inflammatory Index, DII®., Results: Energy-restricted anti-inflammatory diet resulted in significant weight loss (-7.1%, p  < 0.001), in reducing the visceral adiposity (-22.3%, p  < 0.001), metabolic (HOMA-IR, -15.5%; total cholesterol, -5.3%; LDL-C, -4.6%; triglycerides, -12.2%), and inflammatory biomarkers (hs-CRP, -29.5%; IL-6, -18.2%; TNF- α , -34.2%), with significant improvement of liver parameters (NAFLD-FLS, -143.4%; FLI, -14.3%; FIB-4, -2.5%)., Conclusion: The study showed the effectiveness of the anti-inflammatory diet with significant improvement of liver parameters in younger adults with obesity, which may reinforce the effectiveness of nutrition-based lifestyle programs, with an anti-inflammatory dietary approach for the treatment and resolution of NAFLD., Competing Interests: Dario Rahelić is the director of Vuk Vrhovac University Clinic for Diabetes, Endocrinology and Metabolic Diseases at Merkur University Hospital, Zagreb, Croatia. He is the president of Croatian Society for Diabetes and Metabolic Disorders of Croatian Medical Association. He serves as an Executive Committee member of Croatian Endocrine Society, Croatian Society for Obesity and Croatian Society for Endocrine Oncology. He was a board member and secretary of IDF Europe and currently, he is the chair of IDF YLD Program. He has served as an Executive Committee member of Diabetes and Nutrition Study Group of EASD and currently, he serves as an Executive Committee member of Diabetes and Cardiovascular Disease Study Group of EASD. He has served as principal investigator or co-investigator in clinical trials of AstraZeneca, Eli Lilly, MSD, Novo Nordisk, Sanofi Aventis, Solvay, and Trophos. He has received honoraria for speaking or advisory board engagements and consulting fees from Abbott, Amgen, AstraZeneca, Bauerfeund, Bayer, Boehringer Ingelheim, Eli Lilly, Lifescan–Johnson and Johnson, Novartis, Novo Nordisk, MSD, Merck Sharp and Dohme, Mylan, Pfizer, Pliva, Roche, Salvus, Sanofi Aventis, and Takeda. Sanja Klobučar Majanović is the Vice President of Croatian Society for Diabetes and Metabolic Disorders of Croatian Medical Association and the Vice President of Croatian Society for Obesity of Croatian Medical Association. She serves as an Executive Committee member of Croatian Endocrine Society. She has served as principal investigator or co-investigator in clinical trials of Eli Lilly, MSD, and Sanofi Aventis. She has received honoraria for speaking or advisory board engagements and consulting fees from Abbott, AstraZeneca, Boehringer Ingelheim, Eli Lilly, Lifescan–Johnson and Johnson, Novartis, Novo Nordisk, MSD, Merck, Sharp and Dohme, Mylan, Pliva, and Sanofi Aventis. Other authors declare that they have no conflicts of interest., (Copyright © 2021 Gordana Kenđel Jovanović et al.)

Objectives: Radiological and clinical cerebral vasospasm (CV) is defined either as a delayed narrowing of cerebral arteries after aneurysmal subarachnoid hemorrhage (aSAH) or/and occurrence of new neurological deficit/worsening of Modified Glasgow coma score for 2 or more points. The objective of this study is to determine the presence and correlation between clinical and radiological presence of vasospasm in patients with aSAH., Methods: This study was designed as a clinical, prospective single center study at the Clinic of Neurosurgery, Clinical Center of Vojvodina, Novi Sad, Serbia. A total of 50 patients was included in the study after having radiologically confirmed aSAH. Intensity and region of CV was determined by CT and CTA performed both on admission and on day 9 of hospitalization, except for cases where clinical protocol required earlier imaging due to occurrence of clinical signs and symptoms of CV. In all patients, values of arterial blood pressure (PABP), headache (HA), body temperature (PBT), nonspecific behaviors (NSB), deterioration of consciousness (DC), new neurological deficit (NND), deterioration of two points or more per modified Glasgow Coma Scale (DmGCS ≥ 2) were monitored., Results: CTA showed angiographic vasospasm detected in 100% patients with aSAH. Statistically significant positive correlation was found between the intensity of radiological CV and appearance of NND and DmGCS ≥ 2., Conclusions: This study confirms that CV always follows aSAH. Future research into pathophysiology of CV is needed in order to determine exact treatment strategies and targets so treatment towards zero mortality can be achieved.

To test the effect of the moment of bentonite addition, co-addition of tannins, and bentonite type on the differential affinity of pathogenesis-related (PR) proteins towards bentonite during grape must fermentation, three separate experiments were set up. PR proteins in the obtained wines were analyzed by reverse phase and size exclusion high-performance liquid chromatography (HPLC). The most significant reduction of bentonite dose and PR protein concentration was achieved by applying bentonite in the last third of fermentation. Particular thaumatin-like proteins (TLP) and proteins with lower molecular mass in general were more affected than others, while TLPs were more affected than chitinases. Exogenous enological tannins interacted with particular PR proteins, mostly TLPs, and lowered the total bentonite dose required. The combined application of tannins and bentonite in fermentation removed more PR proteins than bentonite alone, but did not achieve a synergistic effect in reducing the bentonite dose. Various bentonite types, including two Na-activated bentonites, an activated Na bentonite with specifically adsorbed silica, and an active Na-Ca bentonite, exhibited differential affinity towards different PR proteins. The results obtained could be used in developing wine fining protocols which combine treatments with complementary affinity for adsorption and removal of PR proteins, and in this way achieve greater efficiency of bentonite fining by reducing its total dose, which is of significant interest to the wine industry.

To test the effects of bentonite addition at various stages of fermentation, five Malvazija istarska white grape must vinification treatments were performed with 100 g/hL of bentonite added in clear juice, at the beginning, in the middle, and at the end of fermentation, while control was fermented without bentonite. Phenols and free and bound volatile aromas were determined by HPLC-DAD and SPE-GC-MS. Wines were evaluated sensorially. Fining during fermentation reduced the total bentonite dose required, and was most effective near the end of fermentation with the reduction of 16% and 21%, depending on the protein stability test. All treated wines preserved more hydroxycinnamoyltartaric acids with respect to control. The side-effect of these treatments on varietal aromas was moderate, but enhanced the preservation of key fermentation volatiles in relation to control, and exhibited positive sensory effects. It was concluded that bentonite added during fermentation may positively affect wine quantity and quality., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Acne vulgaris is a common chronic skin disorder of the pilosebaceous unit with a wide range of clinical presentations, which depend on the age of onset of acne, sex, family history of acne, and genetic factors, especially the genes affecting keratinization and desquamation. This retrospective study investigated pediatric acne using the patients' past medical history, with patients aged from newborns to 15 years of age. Acne were further stratified by 5 parameters: sex, age, family history, acne type, and localization. Our main aim was to investigate the possible association between selected parameters and the presence or absence of family history of acne. We did not find statistically significant correlation between sex, age of onset, and positive family history of acne. Furthermore, we did not find any association between age of onset and family history according to family members (mother/father/brother or sister). However, we found statistically significant correlation between sex and type of acne. This retrospective analysis of pediatric acne in Croatia did not reveal statistically significant correlation between positive family history and sex, age of onset, and clinical type of acne. In analyzing the correlation between family history and localization of acne, however, we found that the number of patients with acne localized on both the face and trunk and positive family history was statistically significant higher than expected.

Background: High resolution melting (HRM) analysis is one of the newer, reliable, and sensitive genotyping techniques, which offers considerable time and cost savings. P-selectin is an adhesion molecule that has a role in the initial phases of leukocyte adhesion to stimulated platelets and endothelial cells in inflammation. Multiple polymorphisms in P-selectin gene (SELP) that affect the protein sequence have been described. The aim of this study was to design, optimize, and validate a simple and rapid in-house HRM-based method for genotyping the NM&#95;003005.3:c.992G>A (c.992G>A), NM&#95;003005.3:c.1918G>T (c.1918G>T), and NM&#95;003005.3:c.2266A>C (c.2266A>C) SELP polymorphisms., Methods: Initial genotyping of three SELP polymorphisms was performed by applying polymerase chain reaction (PCR) with sequence-specific primers (SSP), which was used as a reference method for determination of analytical sensitivity. PCR-HRM was performed with primers for c.2266A>C reported in the literature. Primers for the remaining two polymorphisms were designed using Primer-BLAST. Precision testing was performed using three samples with different genotypes. For accuracy, analytical sensitivity and specificity testing, 20 wild type, 10 heterozygous, and 10 homozygous samples were chosen per polymorphism. Results were expressed as percentage of concordance with the acceptability criterion ≥95%., Results: Agreement of results was 100% for all validation parameters except for analytical sensitivity for c.1918G>T and c.2266A>C, with agreement of 90%. Repeated analysis using both methods revealed an error in initial genotyping and correct genotyping by PCR-HRM, which was confirmed by Sanger sequencing., Conclusion: The validation confirmed PCR-HRM as a precise, accurate, and specific method for genotyping the c.992G>A, c.1918G>T, and c.2266A>C SELP polymorphisms., (© 2018 Wiley Periodicals, Inc.)

Introduction: Thrombosis is the most common complication in Philadelphia chromosome negative (Ph-) myeloproliferative neoplasms patients., Patients and Methods: In a cohort of 258 Ph- myeloproliferative neoplasm patients, the difference between patients with and without thrombosis was analyzed according to genetic thrombophilia factors, JAK2 V617F status and burden allele, blood count, cardiovascular risk factors and age. Patients were also divided in polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF) subgroups as well as by the type of thrombosis., Results: Analysis of cardiovascular risk factors regarding arterial thrombosis showed that PV patients with thrombosis had higher incidence of diabetes (P = .030), ET patients more often had hypertension (P = .003) and hyperlipidemia (P = .005), while PMF patients had hyperlipidemia (P = .046) and at least one cardiovascular risk factor (P = .044). Moreover, leukocytes > 18 × 10 9 /L and V617F burden allele > 25.7% were statistically significantly different in PV patients (P = .019 and borderline significant at P = .055, respectively), while in ET patients leukocytes > 9.2 × 10 9 /L (P < .001) and age at diagnosis of > 55 years were statistically significantly different (P = .002). PMF patients with V617F burden allele ≤ 34.8% were more prone to thrombosis (P = .032). When comparing patients with and without venous thrombosis, cutoff value of V617F burden allele > 90.4% was significant for PV patients with thrombosis (P = .036), as was > 56.7% for PMF patients with thrombosis (P = .046). Platelets ≤ 536 × 10 9 /L and age at diagnosis > 54 years showed statistically significant difference for ET patients with thrombosis (P = .015 and P = .041, respectively)., Conclusion: On the basis of our results, a new scoring system for thrombosis risk in PV could be made, while PMF prognostic model may be expanded for better recognition of potential thrombotic risk factors., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC × GC-TOF-MS) was combined with conventional mono-dimensional GC-MS to differentiate Croatian virgin olive oils (VOO) according to variety and geographical origin, based on the profile of volatile aroma compounds isolated by HS-SPME. More than 1000 compounds were detected after untargeted profiling and 256 were identified or tentatively identified, providing one of the most detailed profiles of volatile aroma compounds in VOO up to date. Among them, 131 volatile compounds were significantly different across monovarietal VOOs, while 60 were found useful for the discrimination according to geographical origin. Many major lipoxygenase and minor non-lipoxygenase-generated compounds were shown to have discriminating ability with respect to both factors. Multivariate statistical analysis extracted twenty-one volatile markers with the highest discriminant power for varietal differentiation. The approach reported may have practical application in better understanding, defining, managing, and communicating the varietal or geographical typicity of monovarietal VOOs., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Objectives: Serum chitotriosidase activity (CHIT1) is a biomarker of macrophage activation with an important role in inflammation-induced tissue remodeling and fibrosis. Macrophages have been described to play a crucial role in regulating pathological erythropoiesis in polycythemia vera (PV). The aim of this study was to evaluate CHIT1 in patients diagnosed with Philadelphia-negative myeloproliferative neoplasms (MPNs)., Methods: Using fluorometric assay, we measured CHIT1 in 28 PV, 27 essential thrombocythemia (ET), 17 primary myelofibrosis (PMF), 19 patients with secondary myelofibrosis and in 25 healthy controls., Results: CHIT1 was significantly higher in PV (p < .001) and post-PV myelofibrosis (MF) transformation (post-PV MF) (p = .020), but not in ET (p = .080), post-ET MF transformation (p = .086), and PMF patients (p = .287), when compared to healthy controls. CHIT1 in PV was positively correlated with hemoglobin (p = .026), hematocrit (p = .012), absolute basophil count (p = .030) and the presence of reticulin fibrosis in the bone marrow (p = .023)., Discussion: A positive correlation between CHIT1 and these distinct laboratory PV features might imply macrophages closely related to clonal erythropoiesis as cells of CHIT1 origin. In addition, a positive association between CHIT1 and reticulin fibrosis might indicate its potential role in PV progression., Conclusion: CHIT1 might be considered as a circulating biomarker in PV. Additional studies are needed to clarify the role of CHIT1 in promoting disease progression and bone marrow fibrosis in PV.

To investigate if the anecdotal varietal typicity concepts for two monovarietal virgin olive oils (VOO) can be scientifically supported and explained, VOO samples were sorted according to typicity grade and related to sensory and compositional data for volatiles and phenols. Clear inter-varietal, as well as intra-varietal boundaries between more and less typical samples were established by univariate and multivariate statistics. A variety-typical complex odor was related to a complex VOO volatile composition with relatively high amounts of lipoxygenase-derived compounds, accompanied by hydrocarbons, terpenes, and benzenoids. A simpler odor, described mostly as green and typical for the other monovarietal VOO, coincided with the lower amounts of the same volatiles. The intensity of bitterness and pungency was mostly in correlation with the amount of major phenols. Two monovarietal VOOs were clearly distinguished with respect to their variety-typical expression of pungency: the one described as gradually increasing was tentatively related to pinoresinol, while the other, described as immediately strong and burning, was assumed to originate mainly from p-HPEA-EDA. Intra-varietal typicity grades correlated with hedonic liking, as well as with general VOO positive sensory attributes related to key volatiles and phenols., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: To analyze the frequency of atypical fluorescence in situ hybridization signal patterns and estimate the complete cytogenetic response (CCyR) and major molecular response (MMR) during 12 months of tyrosine kinase inhibitor therapy in patients with newly diagnosed chronic myeloid leukemia., Methods: The study included bone marrow and peripheral blood samples from 122 patients with newly diagnosed chronic myeloid leukemia. Detection of the breakpoint cluster region - Abelson fusion gene ( BCR-ABL1 ) was performed using fluorescence in situ hybridization with a dual-color dual-fusion translocation probe, and MMR analysis was performed using the real-time quantitative polymerase chain reaction method., Results: Variant translocation was determined in 10 samples and a deletion on the derivative chromosome 9 (del/der(9)) was found in 20 samples. The rates of CCyR and MMR were similar between patients with reciprocal translocation, variant translocation, deletion of derivative BCR , or ABL1-BCR fusion gene. The Kaplan-Meier test did not show any significant differences in the rates of CCyR and MMR among those groups of patients., Conclusion: The frequencies of variant translocation and del/der(9) in the present study agree with the results of other studies performed worldwide. No differences were observed in the rates of CCyR and MMR between patients with atypical patterns and reciprocal translocation., Competing Interests: Authors' Disclosures of Potential Conflicts of Interest: No potential conflicts of interest relevant to this article were reported.

Despite the identification of a wide range of inherited and acquired risk factors for arterial ischemic stroke (AIS) in children, genetic risk factors are incompletely characterized and may vary among different populations. We investigated the role of individual and combined inherited prothrombotic and intermediate-risk factors in 73 children with perinatal (n = 35) and childhood AIS (n = 38) and 100 age- and sex-matched controls. Ten polymorphisms in 8 candidate genes encoding coagulation and fibrinolytic proteins (factor V [FV] Leiden, FV HR2, factor II [FII] G20210A, β-fibrinogen [β-FBG]-455G>A, factor XIII [FXIII]-A p.Val34Leu, plasminogen activator inhibitor 1 4G/5G), homocysteine metabolism (methylenetetrahydrofolate reductase [MTHFR] C677T, MTHFR A1298C), and intermediate-risk factors (angiotensin-converting enzyme I/D, apoE ∊2-4) were detected using a multilocus genotyping assay. Allele-specific polymerase chain reaction was used for the determination of human platelet alloantigens (HPA-1, HPA-2, HPA-3, and HPA-5). Factor V Leiden was associated with an increased risk of AIS (odds ratio [OR]: 4.72, 95% confidence interval [CI]: 1.22-18.27) and perinatal AIS (OR: 8.29, 95% CI: 1.95-35.24). Human platelet antigen-3b allele carriers had a 2-fold lower risk of AIS (OR: 0.51, 95% CI: 0.26-0.98) and perinatal AIS (OR: 0.40, 95% CI: 0.18-0.92). A 2.21-fold increased risk of childhood AIS (95% CI: 1.03-4.73) was identified in FXIII-A Leu34 allele carriers. Combined FV Leiden/FV HR2, FV Leiden/MTHFR A1298C, FV Leiden/MTHFR C677T/MTHFR A1298C, and FV Leiden/FV HR2/MTHFR A1298C heterozygosity was identified in children with AIS but not in controls, which revealed a statistically significant difference. This case-control study shows that besides already documented association between FV Leiden and AIS, other previously unreported polymorphisms (FXIII-A p.Val34Leu, HPA-3) and several genotype combinations that always include heterozygous FV Leiden can be related to AIS in Croatian population.

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.