Search

Spray-drying is a common technique for the microencapsulation of bioactive compounds, which is crucial for improving their stability and bioavailability. In this study, the encapsulation efficiency ( EE ), physicochemical properties and in vitro bioaccessibility of phenolic compounds from spray-dried encapsulated phenol-rich extracts of grape pomace, a winery waste, were evaluated. Sodium alginate alone (SA) or in a mixture with gum Arabic (SA-GA) or gelatin (SA-GEL) was used as a coating. SA-GEL achieved the highest EE (95.90-98.01%) and outperformed the intestinal release of phenolics by achieving a bioaccessibility index ( BI ) for total phenolic compounds of 37.8-96.2%. The release mechanism of phenolics from the microcapsules adhered to Fickian diffusion. Encapsulation significantly improved the BI of individual phenolics, with the highest BI values for gallocatechin gallate (2028.7%), epicatechin gallate (476.4%) and o -coumaric acid (464.2%) obtained from the SA-GEL microcapsules. Structural analysis confirmed amorphous matrices in all systems, which improved solubility and stability. These results suggest that encapsulation by spray-drying effectively protects phenolics during digestion and ensures efficient release in the intestine, which improves bioaccessibility. This study contributes to the understanding of biopolymer-based encapsulation systems, but also to the valorisation of grape pomace as a high-value functional ingredient in sustainable food processing.

Background: The SNP rs855791 has been linked to increased hepcidin levels, variations in serum iron, transferrin saturation and red blood cell indices. Our goal was to determine the prevalence of this polymorphism among COPD patients and to assess its impact on iron status parameters in patients with stable COPD., Methods: We analysed iron status parameters and genetic data from 29 COPD patients with wild-type genotype (WT group) and 65 COPD patients with either homozygous or heterozygous genotype (HH group). Additionally, the prevalence of SNP rs855791 was assessed in 192 volunteers., Results: The frequency distribution of SNP rs855791 was comparable between the COPD patients and control subjects (p=0.791). Iron status parameters were within their respective reference values and showed neither statistically nor clinically significant difference between the WT and HH group of COPD patients. However, after excluding patients with (sub)clinical vitamin B12 deficiency and/or hypoxemia, WT group of patients exhibited significantly lower erythropoietin levels (p=0.015). The area under the curve for erythropoietin was 0.688 (95% CI: 0.545-0.830, p=0.015), with an optimal cut-off of 9.74, sensitivity of 61.2% (95% CI: 58.1-64.3) and specificity of 65.0% (95% CI: 61.8-68.3)., Conclusions: Iron status parameters do not differ between WT and HH groups of stable COPD patients. Statistical but not clinical difference in EPO levels was observed in a subgroup of patients. In addition to promoting erythropoiesis, EPO may regulate hepcidin levels and thus influence the development of iron deficiency and/or anaemia. Also, EPO's direct effect on immune cells and down-regulation of inflammatory reactions should be considered in this context., Competing Interests: All the authors declare that they have no conflict of interest in this work.Conflict of Interest: The authors stated that they have no conflicts of interest regarding the publication of this article., (2025 Marko Trtica, Ivana Novaković, Violeta Dopsaj, Branislava Milenković, Jelena Janković, Sanja Dimić-Janjić, Vesna Dopuđa-Pantić, Jelena Martinović, Snežana Jovičić, published by CEON/CEES.)

Most infectious diseases of the gastrointestinal tract can easily be treated by exploiting the already available antibiotics with the change in administration approach and delivery system. Ciprofloxacin (CIP) is used as a drug of choice for many bacterial infections; however, long-term therapy and off-site drug accumulation lead to an increased risk of tendinitis and peripheral neuropathy. To overcome this issue, nanotechnology is being exploited to encapsulate antibiotics within polymeric structures, which not only facilitates dose maintenance at the infection site but also limits off-site side effects. Here, sodium alginate (SA) and thiol-anchored chitosan (TC) were used to encapsulate CIP via a calcium chloride (CaCl 2 ) cross-linker. For this purpose, the B-390 encapsulator was employed in the preparation of nanobeads using a simple technique. The hydrogel-like sample was then freeze-dried, using trehalose or mannitol as a lyoprotectant, to obtain a fine dry powder. Design of Experiment (DoE) was utilized to optimize the nanobead production, in which the influence of different independent variables was studied for their outcome on the polydispersity index (PDI), particle size, zeta potential, and percentage encapsulation efficiency (% EE). In vitro dissolution studies were performed in simulated saliva fluid, simulated gastric fluid, and simulated intestinal fluid. Antibacterial and anti-inflammatory studies were also performed along with cytotoxicity profiling. By and large, the study presented positive outcomes, proving the advantage of using nanotechnology in fabricating new delivery approaches using already available antibiotics.

The phenols from grape pomace have remarkable beneficial effects on health prevention due to their biological activity, but these are often limited by their bioaccessibility in the gastrointestinal tract. Encapsulation could protect the phenolics during digestion and influence the controlled release in such an intestine where their potential absorption occurs. The influence of freeze-drying encapsulation with sodium alginate (SA) and its combination with gum Arabic (SA-GA) and gelatin (SA-GEL) on the encapsulation efficiency ( EE ) of phenol-rich grape pomace extract and the bioaccessibility index ( BI ) of phenolics during simulated digestion in vitro was investigated. The addition of a second coating to SA improved the EE , and the highest EE was obtained with SA-GEL (97.02-98.30%). The release of phenolics followed Fick's law of diffusion and the Korsmeyer-Peppas model best fitted the experimental data. The highest BI was found for the total phenolics (66.2-123.2%) and individual phenolics (epicatechin gallate 958.9%, gallocatechin gallate 987.3%) using the SA-GEL coating were used. This study shows that freeze-dried encapsulated extracts have the potential to be used for the preparation of various formulations containing natural phenolic compounds with the aim of increasing their bioaccessibility compared to formulations containing non-encapsulated extracts.

Both grape pomace and whey are waste products from the food industry that are rich in valuable ingredients. The utilization of these two by-products is becoming increasingly possible as consumer awareness of upcycling increases. The biological activities of grape pomace extract (GPE) are diverse and depend on its bioavailability, which is influenced by processes in the digestive system. In this work, goat whey protein (GW) was used as the primary coating to protect the phenolic compounds of GPE during the spray drying process. In addition, trehalose (T), sucrose (S), xylose (X), and maltodextrin (MD) were added to the goat whey proteins as co-coatings and protein stabilizers. All spray drying experiments resulted in microcapsules (MC) with a high encapsulation efficiency (77.6-95.5%) and yield (91.5-99.0%) and almost 100% recovery of phenolic compounds during the release test. For o -coumaric acid, the GW-coated microcapsules (MC) showed a bioavailability index of up to 731.23%. A semi-crystalline structure and hydrophilicity were characteristics of the MC coated with 10% T, S, X, or 5% MD. GW alone or in combination with T, S, MD, or X proved to be a promising carrier for polyphenols from grape pomace extract and ensured good bioavailability of these natural antioxidants.

While the effects of chronic exposure to microplastic particles (MPs) are extensively studied, the outcomes of a single treatment have received relatively less attention. To investigate MPs' potential acute toxicity, including their impact on general health status (victual consumption, sensorimotor deficits, and clinical toxicity signs) and serum biochemical parameters (markers of organ/tissue function and oxidative stress indicators), we administered thoroughly characterized MPs (1.4, 35, or 125 mg/kg), generated from polyethylene terephthalate (PET) bottles, to adult male Wistar rats via oral gavage. The MPs' short-term effects were assessed with well-established tests and methods. The results point to the absence of sensorimotor deficits and clinical toxicity signs, while levels of markers of liver, heart, and kidney function were altered in all MP groups. Decreased victual consumption and increased levels of oxidative stress indicators were evident following treatment with the two higher MP doses. Presented data indicate that examined MPs are able to initiate the development of local changes in tissues and organs within a short time frame, potentially leading to their damage and dysfunction. This study may increase the awareness of the detrimental effects of plastic contamination, as even a single exposure to MPs may provoke adverse health outcomes.

Grape pomace is a byproduct of wineries and a sustainable source of bioactive phenolic compounds. Encapsulation of phenolics with a well-chosen coating may be a promising means of delivering them to the intestine, where they can then be absorbed and exert their health-promoting properties, including antioxidant, anti-inflammatory, anticancer, cardioprotective, and antimicrobial effects. Ionic gelation of grape pomace extract with natural coatings (sodium alginate and its combination with maltodextrins, gelatin, chitosan, gums Tragacanth and Arabic) was performed, and the resulting hydrogel microbeads were then air-, vacuum-, and freeze-dried to prevent spoilage. Freeze-drying showed advantages in preserving the geometrical parameters and morphology of the microbeads compared to other drying techniques. A good relationship was found between the physicochemical properties of the dried microbeads and the in vitro release of phenolics. Freeze-dried microbeads showed the highest cumulative release of phenols in the intestinal phase (23.65-43.27 mg GAE /g MB ), while the most suitable release dynamics in vitro were observed for alginate-based microbeads in combination with gelatin, gum Arabic, and 1.5% ( w / v ) chitosan. The results highlight the importance of developing encapsulated formulations containing a natural source of bioactive compounds that can be used in various functional foods and pharmaceutical products.

Grape pomace is a by-product of winemaking characterized by a rich chemical composition from which phenolics stand out. Phenolics are health-promoting agents, and their beneficial effects depend on their bioaccessibility, which is influenced by gastrointestinal digestion. The effect of encapsulating phenol-rich grape pomace extract (PRE) with sodium alginate (SA), a mixture of SA with gelatin (SA-GEL), and SA with chitosan (SA-CHIT) on the bioaccessibility index ( BI ) of phenolics during simulated digestion in vitro was studied. A total of 27 individual phenolic compounds (IPCs) were quantified by UHPLC. The addition of a second coating to SA improved the encapsulation efficiency ( EE ), and the highest EE was obtained for SA-CHIT microbeads (56.25%). Encapsulation affected the physicochemical properties (size, shape and texture, morphology, crystallinity) of the produced microbeads, which influenced the delivery of phenolics to the intestine and their BI . Thus, SA-GEL microbeads had the largest size parameters, as confirmed by scanning electron microscopy (SEM), and the highest BI for total phenolic compounds and IPCs (gallic acid, 3,4-dihydroxybenzoic acid and o -coumaric acid, epicatechin, and gallocatechin gallate) ranged from 96.20 to 1011.3%. The results suggest that encapsulated PRE has great potential to be used as a functional ingredient in products for oral administration.

Grape pomace is a sustainable source of bioactive phenolic compounds used in various industries. The recovery of phenolic compounds could be improved by biological pretreatment of grape pomace, as they are released from the lignocellulose structure by the activity of the enzymes produced. The influence of grape pomace pretreatment with Rhizopus oryzae under solid-state conditions (SSF) on the phenolic profile and chemical composition changes was studied. SSF was performed in laboratory jars and in a tray bioreactor for 15 days. Biological pretreatment of grape pomace resulted in an increase in the content of 11 individual phenolic compounds (from 1.1 to 2.5-fold). During SSF, changes in the chemical composition of the grape pomace were observed, including a decrease in ash, protein, and sugar content, and an increase in fat, cellulose, and lignin content. A positive correlation ( r > 0.9) was observed between lignolytic enzymes and the hydrolytic enzyme's xylanase and stilbene content. Finally, after 15 days of SSF, a weight loss of GP of 17.6% was observed. The results indicate that SSF under experimental conditions is a sustainable bioprocess for the recovery of phenolic compounds and contributes to the zero-waste concept by reducing waste.

Grape pomace is a byproduct of wineries and a rich source of phenolic compounds that can exert multiple pharmacological effects when consumed and enter the intestine where they can then be absorbed. Phenolic compounds are susceptible to degradation and interaction with other food constituents during digestion, and encapsulation may be a useful technique for protecting phenolic bioactivity and controlling its release. Therefore, the behavior of phenolic-rich grape pomace extracts encapsulated by the ionic gelation method, using a natural coating (sodium alginate, gum arabic, gelatin, and chitosan), was observed during simulated digestion in vitro. The best encapsulation efficiency (69.27%) was obtained with alginate hydrogels. The physicochemical properties of the microbeads were influenced by the coatings used. Scanning electron microscopy showed that drying had the least effect on the surface area of the chitosan-coated microbeads. A structural analysis showed that the structure of the extract changed from crystalline to amorphous after encapsulation. The phenolic compounds were released from the microbeads by Fickian diffusion, which is best described by the Korsmeyer-Peppas model among the four models tested. The obtained results can be used as a predictive tool for the preparation of microbeads containing natural bioactive compounds that could be useful for the development of food supplements.

Background: Colorectal carcinoma is one of the most commonly diagnosed malignancies worldwide. Consumption of dietary supplements and nutraceuticals such as phenolic compounds may help combat colorectal carcinoma. The effect of two phenolic-rich extracts prepared from biotransformed grape pomace on the antioxidant properties and antiproliferative activity against two colorectal cancer cell lines (Caco-2 and SW620) were investigated., Methods: A 15-day solid-state fermentation with the white-rot fungi Phanerochaete chrysosporium and Trametes gibbosa was used to biotransform grape pomace. Solid-liquid extraction was then performed to extract bioactive compounds. The extract was analyzed for the determination of phenolic compounds by ultra-high performance liquid chromatography and in vitro assays of biological activities (antioxidant activity, antiproliferative activity, cell cycle analysis)., Results: The 4 days of solid-state fermentation proved to be the optimal period to obtain the maximum yield of phenolic compounds. The tested extracts showed significant antioxidant and antiproliferative activities. Grape pomace treated with P. chrysosporium and T. gibbosa reduced cancer cell growth by more than 60% at concentrations (solid/liquid ratio) of 1.75 mg/mL and of 2.5 mg/mL, respectively. The cell cycle perturbations induced by the grape pomace extracts resulted in a significant increase in the number of cells in the S (9.8%) and G2/M (6.8%) phases of SW620 exposed to T. gibbosa after 48 hours, while P. chrysosporium increased the percentage of cells in the G1 phase by 7.7%. The effect of grape pomace extracts on Caco-2 was less pronounced., Conclusions: The obtained results suggest the presence of bioactive compounds in biotransformed grape pomace as a residue from winemaking, which could be used to prevent colon cancer., (© 2023. The Author(s).)

This paper presents quantitative research results regarding a predictive model of pandemic disaster fear caused by the coronavirus disease (COVİD-19). The aim of this paper was to establish the level and impact of certain demographic and socioeconomic characteristics on pandemic disaster fear caused by the coronavirus (COVID-19). The research was conducted using a questionnaire that was provided and then collected online for 1226 respondents during May 2021. A closed, five-point Likert scale was used to create the structured questionnaire. The first section of the questionnaire included research questions about the participants' socioeconomic and demographic characteristics, while the second section included issue questions about fear caused by COVID-19. The results of multivariate regression analyses showed the most important predictor for fear of COVID-19 to be gender, followed by age and education level. Furthermore, the results of t -tests showed statistically significant differences between men and women in terms of different aspects of pandemic disaster fear caused by the coronavirus disease. Our results have several significant public health implications. Women who were more educated and knowledgeable, married, and older, reported a greater fear of the outbreak at various levels. Decision-makers can use these findings to identify better strategic opportunities for pandemic disaster risk management.

Sustained activation of pro-apoptotic signaling due to a sudden and prolonged disturbance of cerebral blood circulation governs the neurodegenerative processes in prefrontal cortex (PFC) of rats whose common carotid arteries are permanently occluded. The adequate neuroprotective therapy should minimize the activation of toxicity pathways and increase the activity of endogenous protective mechanisms. Several neuroprotectants have been proposed, including progesterone (P 4 ). However, the underlying mechanism of its action in PFC following permanent bilateral occlusion of common carotid arteries is not completely investigated. We, thus herein, tested the impact of post-ischemic P 4 treatment (1.7 mg/kg for seven consecutive days) on previously reported aberrant neuronal morphology and amount of DNA fragmentation, as well as the expression of progesterone receptors along with the key elements of Akt/Erk/eNOS signal transduction pathway (Bax, Bcl-2, cytochrome C, caspase 3, PARP, and the level of nitric oxide). The obtained results indicate that potential amelioration of histological changes in PFC might be associated with the absence of activation of Bax/caspase 3 signaling cascade and the decline of DNA fragmentation. The study also provides the evidence that P 4 treatment in repeated regiment of administration might be effective in neuronal protection against ischemic insult due to re-establishment of the compromised action of Akt/Erk/eNOS-mediated signaling pathway and the upregulation of progesterone receptors.

Background: Different variables determine the performance of cyclists, which brings up the question how these parameters may help in their classification by specialty. The aim of the study was to determine differences in cardiorespiratory parameters of male cyclists according to their specialty: flat riders (N.=21), hill riders (N.=35), or sprinters (N.=20) and obtain the multivariate model for further cyclists classification by specialties, based on selected variables., Methods: Seventeen variables were measured at submaximal and maximum load on the cycle ergometer Cosmed E 400HK (Cosmed, Rome, Italy) (initial 100 W with 25-W increase, 90-100 rpm). Multivariate discriminant analysis was used to determine which variables group cyclists within their specialty, and to predict which variables can direct cyclists to a particular specialty., Results: Among nine variables that statistically contribute to the discriminant power of the model, achieved power on the anaerobic threshold and the produced CO2 had the biggest impact. The obtained discriminatory model correctly classified 91.43% of flat riders, 85.71% of hill riders, while sprinters were classified completely correct (100%), i.e. 92.10% of examinees were correctly classified, which point out the strength of the discriminatory model., Conclusions: Respiratory indicators mostly contribute to the discriminant power of the model, which may significantly contribute to training practice and laboratory tests in future.

Background: Extracellular adenine nucleotides and nucleosides, such as ATP and adenosine, are among the most recently identified and least investigated diffusible signaling factors that contribute to the structural and functional remodeling of the brain, both during embryonic and postnatal development. Their levels in the extracellular milieu are tightly controlled by various ectonucleotidases: ecto-nucleotide pyrophosphatase/phosphodiesterases (E-NPP), alkaline phosphatases (AP), ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDases) and ecto-5'- nucleotidase (eN)., Methods: Studies related to the expression patterns of ectonucleotidases and their known features during brain development are reviewed, highlighting involvement of these enzymes in synapse formation and maturation in physiological as well as in pathological states., Results: During brain development and in adulthood all ectonucleotidases have diverse expression pattern, cell specific localization and function. NPPs are expressed at early embryonic days, but the expression of NPP3 is reduced and restricted to ependymal area in adult brain. NTPDase2 is dominant ectonucleotidase existing in the progenitor cells as well as main astrocytic NTPDase in the adult brain, while NTPDase3 is fully expressed after third postnatal week, almost exclusively on varicose fibers. Specific brain AP is functionally associated with synapse formation and this enzyme is sufficient for adenosine production during neurite growth and peak of synaptogenesis. eN is transiently associated with synapses during synaptogenesis, however in adult brain it is more glial than neuronal enzyme., Conclusion: Control of extracellular adenine nucleotide levels by ectonucleotidases are important for understanding the role of purinergic signaling in developing tissues and potential targets in developmental disorders such as autism., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

17β-Estradiol (E2) rapidly, by binding to membrane estrogen receptors, activates cell signaling cascades which induce formation of new dendritic spines in the hippocampus of males as in females, but the interaction with other metabolic processes, such as extracellular adenine nucleotides metabolism, are currently unknown. Extracellular adenine nucleotides play significant roles, controlling excitatory glutamatergic synapses and development of neural circuits and synaptic plasticity. Their precise regulation in the synaptic cleft is tightly controlled by ecto-nucleoside triphosphate diphosphohydrolase (NTPDase)/ecto-5'-nucleotidase (eN) enzyme chain. Therefore, we sought to clarify whether a single systemic injection of E2 in male rats is accompanied by changes in the expression of the pre- and postsynaptic proteins and downstream kinases linked to E2-induced synaptic rearrangement as well as alterations in NTPDase/eN pathway in the hippocampal synaptosomes. Obtained data showed activation of mammalian target of rapamycin and upregulation of key synaptic proteins necessary for spine formation, 24 h after systemic E2 administration. In E2-mediated conditions, we found downregulation of NTPDase1 and NTPDase2 and attenuation of adenine nucleotide hydrolysis by NTPDase/eN enzyme chain, without changes in NTPDase3 properties and augmentation of synaptic tissue-nonspecific alkaline phosphatase (TNAP) activity. Despite reduced NTPDase activities, increased TNAP activity probably prevents toxic accumulation of ATP in the extracellular milieu and also hydrolyzes accumulated ADP due to unchanged NTPDase3 activity. Thus, our initial evaluation supports idea of specific roles of different ectonucleotidases and their coordinated actions in E2-mediated spine remodeling and maintenance.

Nucleoside triphosphate diphosphohydrolase3 (NTPDase3) is membrane-bound ecto-enzyme which hydrolyzes extracellular ATP, thus modulating the function of purinergic receptors and the pattern of purinergic signaling. Here we analyzed the developmental expression of NTPDase3 in female hypothalamus, cerebral cortex and hippocampal formation at different postnatal ages (PD7-PD90) by qRT-PCR and immunohistochemistry. In hypothalamus and hippocampus, a similar developmental profile was seen: NTPDase3 gene expression was stable during postnatal development and increased in adults. In the cortex, upregulation of NTPDase3 mRNA expression was seen at PD15 and further increase was evidenced in adults. Immunohistochemical analysis at PD7 revealed faint neuronal NTPDase3 localization in a dorsal hypothalamus. The immunoreactivity (ir) gradually increased in PD15 and PD20, in clusters of cells in the lateral, ventral and dorsomedial hypothalamus. Furthermore, in PD20 animals, NTPDase3-ir was detected on short fibers in the posterior hypothalamic area, while in PD30 the fibers appeared progressively longer and markedly varicose. In adults, the strongest NTPDase3-ir was observed in collections of cells in dorsomedial hypothalamic nucleus, dorsal and lateral hypothalamus and in several thalamic areas, whereas the varicose fibers traversed entire diencephalon, particularly paraventricular thalamic nucleus, ventromedial and dorsomedial hypothalamic nuclei, the arcuate nucleus and the prefornical part of the lateral hypothalamus. The presumably ascending NTPDase3-ir fibers were first observed in PD20; their density and the varicose appearance increased until the adulthood. Prominent enhancement of NTPDase3-ir in the hypothalamus coincides with age when animals acquire diurnal rhythms of sleeping and feeding, supporting the hypothesis that this enzyme may be involved in regulation of homeostatic functions., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Pregnancy is a stressful condition linked with altered lipid profile, increased oxidative stress and increased inflammation processes. The purpose of the present study was to determine the associations between those alterations with increased weight gain during pregnancy., Methods: The atherogenic index of plasma (AIP) and oxidative stress status parameters were determinated in 50 healthy and 172 pregnant women with non-complicated pregnancy. Pregnant women were divided in four groups according to body mass index (BMI) values (BMI quartiles)., Results: Oxidative stress parameters were significantly lower in the control group compared with all the pregnant women quartiles. Unexpectedly, differences in oxidative stress parameters between BMI quartiles groups were not significant. The antioxidant defence parameters remained quite similar in the different BMI quartiles. Weight gain and paraoxonase-1 (PON1) activities were independently associated with increased AIP while weight gain and triglyceride concentration were found to be significant predictors of PON1 activities., Conclusions: The results of our current study indicate the association of maternal weight gain during pregnancy and altered lipid profile, elevated oxidative stress and changed antioxidative capacity of PON1. Taken together all these facts indicate possible increased risk of cardiovascular disease (CVD) development in later life if the weight gain during pregnancy is excessive.

The objectives of this study were to determine (a) if reactive oxygen metabolites (ROMs) are a reliable parameter for monitoring oxidative stress in athletes alone or in association with other parameters of oxidative stress and depending on whether antioxidant supplements are taken or not; (b) the level of oxidative stress in athletes before the competition season; and (c) if oxidative status could be improved in volleyball athletes. Sixteen women athletes (supplemented group) received an antioxidant cocktail containing vitamin E, vitamin C, zinc gluconate, and selenium as a dietary supplement during a 6-week training period, whereas 12 of them (control group) received no dietary supplement. Blood samples were taken before and after the training period. The following parameters were measured: ROMs, superoxide anion (O2⁻₂), malondialdehyde (MDA), advanced oxidation protein products (AOPP), lipid hydroperoxide (LOOH), biological antioxidative potential (BAP), paraoxonase activity toward paraoxon (POase) and diazoxon (DZOase), superoxide dismutase(SOD), total sulfydryl group concentration (SH groups), and lipid status. Reactive oxygen metabolites were taken as the dependent variable and MDA, O2⁻₂, AOPP, and LOOH as independent variables. In the group of athletes who have received supplementation, linear regression analysis revealed that the implemented model had a lower influence on dROMs (70.4 vs. 27.9%) after the training period. The general linear model showed significant differences between parameters before and after training/supplementation (Wilks' lambda = 0.074, F = 11.76, p < 0.01). At the partial level, significant increases in ROM levels (p <0.05, 95% confidence interval [CI]: 286-337), SOD activity (CI: 113-144), and BAP (CI: 2,388-2,580) (p < 0.01) were observed. The association between ROMs and other parameters of oxidative stress was reduced in athletes who received supplements. During the precompetition training period, treatment with dietary supplements prevented the depletion of antioxidative defense in volleyball athletes.

Aim: The objectives of this study were: 1) to evaluate pro-oxidant-antioxidant balance (PAB) associations with the commonly measured parameters of oxidative stress and antioxidatve defence in elite female volleyball athletes; 2) to investigate changes in the parameters of oxidative stress during a period of intense training and dietary antioxidant supplementation., Methods: Twenty-seven female volleyball players participated in this study. Blood samples were collected the day before the pre-competitive mesocycle training period began. After the first blood sample donation and during the next six weeks fourteen players (supplemented group) received a cocktail of antioxidants while thirteen of them (control group) received no dietary supplementation. The following parameters were measured: reactive oxygen metabolites (ROMs), superoxide anion (O2-), malondialdehyde (MDA), advanced oxidation protein products (AOPP), lipid hydroperoxides (LOOH), biological antioxidative potential (BAP), paraoxonase activity toward paraoxon (POase) and diazoxon (DZOase), superoxide dismutase (SOD), total sulphydryl group concentration (SH groups) and pro-oxidant-antioxidant balance., Results: Significant associations were observed between biomarkers of oxidative damage with PAB in multiple linear regression model in the supplemented and the control groups (82.3% vs. 83.1%) before training and in the control group (82.1%) after training. Significant associations between antioxidative defence parameters and PAB values were found in the supplemented group after six-weeks of training (57%)., Conclusion: In the absence of antioxidant supplementation, PAB values were dependent on the association with biomarkers of oxidative damage before and after training. After a six-week training period and the applied antioxidant supplementation, PAB values were under the influence of non-enzymatic anti-oxidative defence.

Objective: The aim of this study was to evaluate how conditions that precede anaemia (iron store depletion and iron-deficient erythropoiesis) affect human serum paraoxonase PON1 activity., Design and Methods: Based on haemoglobin, transferrin saturation and serum ferritin values 119 athletes were divided into three groups: with iron depletion, with deficient erythropoiesis and controls. The following parameters were measured: paraoxonase activity towards paraoxon (POase) and diazoxon (DZOase), lipid hydroperoxides (LOOH), the pro-oxidant-antioxidant balance (PAB), red blood cells (RBC) and lipid status., Results: Significant differences were found between athletes with different stages of iron deficiency and controls with respect to PON 1 activity and oxidative stress status parameters (Wilks' Lambda=0.712, F=5.241, p<0.001, η(2)=0.156). There was no significant difference between the PON1 192 Q and R polymorphism distribution in the two groups of athletes with different stages of iron deficiency and controls (χ(2)=1.086; p=0.896). PON1 activity was positively correlated with RBCs, haemoglobin, transferrin saturation (p<0.001) and ferritin (p=0.037) and negatively correlated with LOOH (p=0.044) in all three study groups., Conclusions: Deficient erythropoiesis in athletes contributes to impaired PON1 activity. In contrast, iron depletion, regardless of increased oxidative stress, does not affect PON1 activity., (Crown Copyright © 2010. Published by Elsevier Inc. All rights reserved.)

The activity of mitochondrial superoxide dismutase (MnSOD) and cytosol superoxide dismutase (CuZnSOD) was measured in corresponding subcellular fractions prepared from the thymi of intact and chronically gonadectomized (GX) rats of both sexes, as well as of GX male and female rats injected subcutaneously with a single dose of 5 microg estradiol benzoate (EB) and/or 2 mg progesterone (P). Animals were sacrificed 2 h or 24 h following hormone treatment. In the females, the activity of MnSOD in the thymus was stable during the estrous cycle and did not change after ovariectomy. Treatment of GX females with estradiol benzoate resulted 2 h later in a significant elevation of MnSOD activity, whereas 24 h later the activity returned back to control values. On the other hand, treatment of GX females with progesterone had no effect on the MnSOD activity. However, combined hormone treatment, in which EB injection preceded progesterone injection by one hour, enhanced the effect on MnSOD activity similar to that of estradiol benzoate alone. The activity of CuZnSOD in cycling rats was increased in proestrus, whereas removal of the ovaries kept the values at low diestrus and estrus levels. Contrary to MnSOD, CuZnSOD activity did not change after EB treatment of GX females, while progesterone increased the enzyme activity at 2 h and 24 h after hormone treatment. However, combined EB+P treatment proved to be ineffective. In the males, neither MnSOD nor CuZnSOD activity was affected by the removal of testes or by progesterone treatment of GX animals. Only EB injection to GX rats significantly increased CuZnSOD activity 24 h later.

The effects of gonadal steroid hormone, 17beta-estradiol (E2), in vitro on rat brain mitochondria Ca2+ movement were investigated. Intrasynaptosomal mitochondria Ca2+ uptake via an energy-driven Ca2+ uniporter have Km = 112.73 +/- 7.3 micromol x l(-1) and Vmax = 21.97 +/- 1.7 nmol 45Ca2+ mg(-1). Ca2+ release trough a Na+/Ca2+ antiporter was measured with a Km for Na+ of 43.7 +/- 2.6 mmol x l(-1), and Vmax of 1.5 +/- 0.3 nmol 45Ca2+ mg(-1). Addition of estradiol in preincubation mixture did not affect the uptake of Ca2+ mediated by the ruthenium red-sensitive uniporter, while it produced biphasic effect on Na-dependent Ca2+ efflux. Estradiol at concentrations up to 1 nmol x l(-1) decreased the efflux significantly (63% inhibition with respect to the control), and at concentrations above 10 nmol x l(-1) increased it exponentially. The maximum inhibiting concentration of estradiol (0.5 nmol x l(-1)) increased the affinity of the uniporter (Km reduced by about 30%), without affecting significantly the capacity (Vmax) for Na+. The results presented suggest that estradiol inhibits Na-dependent Ca2+ efflux from mitochondria and acts on mitochondrial retention of Ca2+, which may modulate mitochondrial and consequently synaptosomal content of Ca2+, and in this way exerts its role in the homeostasis of calcium in nerve terminals.

The activities of glutathione dependent antioxidant enzymes were measured in subcellular fractions of whole brain homogenates prepared from ovariectomized (OVX) female rats, untreated or treated 2 h or 24 h prior to sacrifice with a single dose of 2 mg progesterone (P) or 5 micrograms estradiol benzoate (EB). Glutathione peroxidase (GSH-Px) activity was not changed following systemic administration of EB, but P increased GSH-Px in the brain of OVX rats 24 h after the treatment. The activity of glutathione reductase (GR) was suppressed by EB short time, only 2 h following treatment, whereas P increased the enzyme activity 24 h after treatment. On the other hand, the activities of catalase (CAT) and glutathione-S-transferase (GST) were not changed following systemic administration of EB or P. The present work was carried out to study the involvement of ovarian steroids, especially P, in the control of GSH-Px and GR activities, and our results suggest that oxidative stress in the brain of female rats may be modulated by the level of progesterone.

In the present study distribution and enzymatic properties of ecto-Mg(2+)-ATPase were determined in synaptic plasma membrane (SPM) preparations isolated from the hippocampus, caudate nucleus and whole brains of female rats. Western blot analysis using anti-ecto-Mg(2+)-ATPase antibody revealed the association of Mg(2+)-ATPase with SPM prepared from all the three brain sources, yet the enzyme was most abundant in caudate nucleus membranes, being 30% and 22% more abundant than in the hippocampal and whole brain tissue SPM, respectively. The evidence is also presented that kinetic properties of the brain Mg(2+)-ATPase are not under the control of circulating sex steroids. It was confirmed that the enzyme is activated by millimolar concentrations of Mg2+ and that it cannot be effectively inhibited by known ATPase inhibitors. The most pronounced differences in kinetic properties observed were 2.5 fold higher apparent affinity for ATP and 59% higher specific activity of Mg(2+)-ATPase of the caudate nucleus as compared with the enzyme from the hippocampus. On the other hand, the apparent enzyme affinity for Mg2+ was almost equal in all SPM preparations tested. Taken together, our results show that ecto-Mg(2+)-ATPase is not uniformly distributed and differs in respect to affinity for ATP in rat brain regions, thus indicating its substantial role in the process of signal transduction via controlling the levels of extracellular ATP.

Membrane vesicles loaded with [Na+], prepared from synaptosomal plasma membranes (SPM) of whole brains (WB), hippocampi (Hip) and caudate nuclei (NC) of female rats, were used to study Na+ -dependent Ca2+ transport across SPM vesicles under the influence of 17beta-estradiol (E2) in vitro. In concentrations near to physiologic, E2 significantly increased 45Ca2+ uptake by SPM vesicles from all the brain tissues investigated. The maximum increase was observed for WB (21%) and Hip (33%) at 10(-9) mol/l, and for NC (31%) at 5 x 10(-9) mol/l of E2. These results (a) confirm our earlier finding that E2 in vitro modulates Na+-dependent Ca2+ transport across synaptosomal membrane in rat brain regions, and (b) suggest Na+/Ca2+ exchange as principal mechanism of the E2-stimulated Na-dependent Ca2+ uptake by membrane vesicles. The involvement of any ATPases as possible mediators is discussed.

The activities and basic enzymatic properties of Na,K-ATPase were examined in synaptosomal plasma membranes (SPM) prepared from rat hippocampus and striatum. A kinetic analysis showed profound differences in apparent affinities for ATP (Km) between hippocampal (1.21 mmol/l) and striatal (0.76 mmol/l) enzyme preparations, as well as in the corresponding Vmax values. However, physiological efficiencies were almost the same. The complex pattern of dose-response curves to ouabain indicated the presence of two high-affinity forms of Na,K-ATPase in the striatum ("very high-": Ki = 3.73 x 10(-8) mol/l and "high-": Ki = 4.21 x 10(-5) mol/l), and one high affinity form in the hippocampus (Ki = 6.6 x 10(-7) mol/l). In addition, both SPM preparations contained one low affinity form with similar Ki. The "very high-affinity" form had positive cooperativity for ouabain inhibition of Na,K-ATPase activity, in contrast to "high" and "low-affinity" forms, which exhibited negative cooperativity. The respective contributions of ouabain-sensitive forms to the total activity were estimated as 22%, 46%, 19% for the striatum and 36%, 45% for the hippocampus. These data clearly demonstrate striking differences in kinetic properties of the hippocampal and striatal Na,K-ATPase that may be due to the isoenzyme diversity and adaptation to specific physiological demands of the examined rat brain regions.

The equilibrium parameters (Bmax and Km) of estradiol binding to synaptosomal plasma membranes prepared from brain regions of adult female rats were examined. Michaelis-Menten plots of the specific binding in samples prepared from the telencephalon and mesencephalic reticular formation showed only one saturation plateau, indicating the presence of a single population of synaptosomal membrane binding sites for estradiol. According to Bmax and Km values, the binding sites appeared to be similar in the two regions. On the other hand, two distinct populations, one with high capacity and lower affinity and another with low capacity and higher affinity, were detected in samples prepared from the nucleus caudatus, brain stem and hippocampus, as shown by two saturation plateaus on corresponding Michaelis-Menten graphs. The equilibrium parameters of the two populations were different in the three regions. The shapes of the Michaelis-Menten plots were substantially the same for intact and for ovariectomized animals. Scatchard and Hill plots revealed cooperativity for estradiol binding in all the regions investigated. These results suggest that central non-genomic effects of estradiol are mediated via multiple types of neuronal membrane binding sites, which appear not to be under ovarian control.

The authors examined the relationship between body mass, body height, haemogram and vital capacity in two groups of pupils exposed to long aeropollution. The first group consisted of children with low vital capacity. In the second group the spirometric test was normal. The authors found differences between these two groups in relation to body mass and hemoglobin concentration.

The article deals with a 16-year old patient who was suffering from bronchial asthma with frequent severe respiratory obstructive crises and decreased values of functional pulmonary tests since his eleventh year. He was on salbutamol and teoline therapy, and since one year and a half on continuous inhalation corticosteroid therapy. During one of the episodes of bronchobstruction when he was on broncholdilatation and oxygen therapy the patient was given novalgetol because of headache. This provoked a grave astmatic attack with loss of consciousness. One and a half year later he was given novalgetol during a dyspneic drisis with headacke. Five minutes later asthatic attack and generalized urticaria appeared as anaphylactic reaction to novalgetol. This is a good example how drugs form the group of amonomethansulphonate (novalgetol), used in analgetic purposes, can induce, like aspirin, asthamtic crisis and anaphylactic reaction. Therefore they should be avoided in asthamtic patients.

The authors followed up the health of 322 elementary-school pupils at Sabac who were exposed to increased air pollution with chemical agents (ammonia, fluor hydrogen, chloracetic acid, sulfur dioxide, soot). The mean annual concentration of these gases were above or insignificantly under maximally permitted values. The results of the examination were as follows: recurrent cough in 12.4 per cent of cases and clinical signs of chronic or obstructive bronchitis in 8.4 per cent of children. Values of vital capacity were under normal in 27.6 per cent of cases and almost in the half of children vital capacity was at the lower normal limit. FEV1/FVC was under normal in 15 per cent of cases and at the lower normal limit in about 18.6 per cent of children. In spite of a great number of children with positive cutaneous tests to inhalation allergens with Prick's method IgE was within normal limits in these children. Thus, a significant noxious effect of allergic components on respiratory organs should be excluded. This was also confirmed by spirometric measurements of the two subsequent maximal expiriums when differences of +/- 3% were registered. However, in children with allergic bronchitis the value dispersion was by 2-3 times greater. Consequently, the authors concluded that damaged respiratory organs in children at Sabac appeared in a greater number of cases than in other places and that these damages were due to increased concentrations of different chemical air pollution agents.

Young adult and elderly male and female intact rats, as well as chronically ovariectomized (OVX) young and elderly female rats, were subjected to an acute stress by cutting the tip of the tail and prolactin (Prl) concentrations were measured in their blood collected by decapitation at various times thereafter. Maximum concentrations of the hormone were markedly lower in all the three groups of elderly rats than those found in the corresponding young animals, and appeared to occur with a delay in the females, but not in the males. In addition, the Prl-response to stress was attenuated in OVX animals regardless of their age. The result of these experiments, performed at two points on the age scale, suggests that in sexually mature rats of both sexes the stress-induced secretion of Prl is inversely related to the age of the animal and that the reverse relationship is retained in OVX females.

The authors present values of immunoglobulins IgA, IgM, IgG and IgE in 40 pupils at Sabac, who were exposed to high aeropollution and in whom spirometric pulmonary functions (vital capacity, FEV1 and FEV1/PVC relation) were under the normal or at its upper limit. Frequent respiratory diseases, sometimes followed by wheezing, were observed in these children. The aim of the study was to detect a possible predisposing factor responsible of these conditions. This was done with the control examinations of immunoglobin values when atopic constitution (IgE) or immunodeficency should be detected. IgG values were, however, within the normal. Thus, the atopic role in the onset of these respiratory disorders was excluded. High-degree aeropollution was cited as the main cause of frequent respiratory diseases and low spirometric values of the pulmonary function.

Prolactin (Prl) secretion in response to an acute stress was studied in ovariectomized (OVX) and/or adrenalectomized (ADX) adult female rats, nontreated or injected sc with a single dose of 5 micrograms oestradiol-17 beta benzoate (OB) or 2 mg progesterone (P). The stress applied consisted of cutting the tip of tail of conscious animals. Radioimmunoassay was used to measure Prl in the serum prepared from blood collected by decapitation 10 min following the stress, i.e., at the point of maximum recorded Prl response. It was found that the capacity of the animals to secrete large quantities of Prl under stress was, when compared to that in intact controls, markedly reduced in OVX or ADX rats and substantially absent in OVX + ADX rats. A 10-fold increase of basal serum Prl, similar in magnitude to the increase in intact controls, was induced by the stress in OVX animals pretreated with OB. On the contrary, pretreatment of OVX animals with P resulted in a complete block of the Prl response to the stress. The stimulative effect of OB was greatly attenuated in stressed OVX + ADX rats. The results suggest that OB potentiates whereas P attenuates the stress-induced secretion of Prl in the female rat, and that the potentiating effect of OB is dependent on functionally intact adrenals.

Thirty-eight children with signs of hydrocephalus have been tested for toxoplasmosis. On the basis of clinical data, including roentgenographical and ophthalmological findings, and serological data (Sabin-Feldman and IgM-IFA tests), congenital toxoplasmosis was confirmed in 15 (39%) infants. The diagnosis was established by demonstration of the persistence of Toxoplasma antibodies. In 13 (34%) infants aged 1 to 3 months there was not enough serological data and these were placed in a group of cases with "suspected congenital toxoplasmosis." Ten children with negative serology for toxoplasmosis and with hydrocephalus were considered not to be infected.

Effects of progesterone (P) and estradiol-17 beta benzoate (EB), applied s.c. into sexually mature, long-term ovariectomized (OVX) rats, on subsequent depolarization-induced uptake of Ca2+ were studied in synaptosomes isolated from the brain stem, mesencephalic reticular formation (MRF), nucleus caudatus putamen (NCP) and the hippocampus. In intact animals, synaptosomal Ca2+ uptake differed from region to region: it was lowest in the brain stem and highest in the hippocampus. In comparison to intact animals, ovariectomy resulted in a marked increase of the uptake regardless the structure investigated, suggesting an inhibitory action of ovaries on the uptake of Ca2+ in a considerable portion of rat brain. Single injection of 2 mg P, given to OVX rats 24 h prior to decapitation, evoked a marked decrease in Ca2+ uptake by synaptosomes of the brain stem and MRF and particularly by those of NCP and the hippocampus. Single injection of 5 micrograms EB into OVX animals 72 h prior to the experiment was as effective as P in inhibiting Ca2+ uptake by synaptosomes of the brain stem and MRF, but less effective than P in case of NCP and the hippocampus. This suggests involvement of P and EB in the modulation of synaptic transmission by affecting neuronal Ca2+ uptake.

In their work authors have analyzed some epidemiologic and etiologic data that can be taken as the elements of risk in leukemogenesis. The investigations have been done retrospectively in 212 children suffering from acute leukemia and treated in Pediatric Clinic in Beograd. Certain factors of risk are particularly studied as: repeated viral infections, repeated use of antibiotics, diagnostic and therapeutic irradiation of children, familiar data on congenital anomalies and cancer and same harmful prezygotic and prenatal influences of possible significance. The authors also presented the results of their cytogenetic investigations obtained from 32 children. It is especially pointed out the significance of detailed data taken from patients suffering from these diseases.