Viruses in shellfish — food safety risks

Shellfish production in the EU has declined in recent decades, which is not the case with global aquaculture production of shellfish. The trend towards a healthy lifestyle and diet is becoming increasingly topical and often involves the consumption of uncooked shellfish. Unfortunately, sh ellfish can often be contaminated with various pathogens, especially virus-es, which can endanger human health. Among the outbreaks of shellfish-borne viruses, the most notable are those caused by Norovirus (NoV) and hepatitis A virus (HAV). However, other viruses belonging to the Herpeviridae, Picornaviridae, Adenoviridae, Astroviridae, and Reoviridae can mainly cause intestinal disease in humans after consumption of contami-nated shellfish. The listed viruses have been detected in shellfish worldwide and they are mostly the consequence of sewage-contaminated water. Numerous preventive and control measures are recommended to solve this problem

Influence of two different culture media on biofilm formation by Listeria monocytogenes isolated from a small-scale meat processing facility

In this study, 20 Listeria monocytogenes isolates detected in a food processing environment and food products were tested for biofilm-forming ability in two different culture media: Tryptone Soya Broth and Luria Bertani Broth. Statistical analysis of the data obtained was performed with the MINITAB software package, version 16.0. The two-sample t-test and confidence interval were used for data analysis. Significant differences between the isolates were observed in the ability to form biofilms

Prolonged Alprazolam Treatment Alters Components of Glutamatergic Neurotransmission in the Hippocampus of Male Wistar Rats—The Neuroadaptive Changes following Long-Term Benzodiazepine (Mis)Use

Alprazolam (ALP), a benzodiazepine (BDZ) used to treat anxiety, panic, and sleep disorders, is one of the most prescribed psychotropic drugs worldwide. The side effects associated with long-term (mis)use of ALP have become a major challenge in pharmacotherapy, emphasizing the unmet need to further investigate their underlying molecular mechanisms. Prolonged BDZ exposure may induce adaptive changes in the function of several receptors, including the primary target, gammaaminobutyric acid receptor type A (GABAAR), but also other neurotransmitter receptors such as glutamatergic. The present study investigated the potential effects of prolonged ALP treatment on components of glutamatergic neurotransmission, with special emphasis on N-Methyl-D-aspartate receptor (NMDAR) in the hippocampus of adult male Wistar rats. The study revealed behavioral changes consistent with potential onset of tolerance and involvement of the glutamatergic system in its development. Specifically, an increase in NMDAR subunits (NR1, NR2A, NR2B), a decrease in vesicular glutamate transporter 1 (vGlut1), and differential modulation of excitatory amino acid transporters 1 and 2 (EAAT1/2, in vivo and in vitro) were observed, alongside a decrease in α1-containing GABAAR following the treatment. By describing the development of compensatory actions in the glutamatergic system, the present study provides valuable information on neuroadaptive mechanisms following prolonged ALP intake

Knowledge of and recommendations for infant safe sleeping among primary health care paediatricians in Split-Dalmatia, Dubrovnik-Neretva and Bjelovar-Bilogora Counties

Cilj: Istražiti poznavanje rizičnih čimbenika za sindrom iznenadne dojenačke smrti od pedijatara primarne zdravstvene zaštite uSplitsko-dalmatinskoj, Dubrovačko-neretvanskoj i Bjelovarsko-bilogorskoj županiji.Ispitanici i metode: Presječnim istraživanjem obuhvaćeni su PPZZ-i u SDŽ-u, DNŽ-u i BBŽ-u u razdoblju od veljače do rujna 2018.godine. Pedijatrima je dostavljen upitnik sa 18 pitanja koja se odnose na mišljenja o spavanju i dojenju, savjete što ih daju roditeljimasvojih bolesnika te znanje o SIDS-u.Rezultati: Analizirano je 29 anketa, a većina ispitanika je starija od 55 godina. Većina pedijatara (72%) tvrdi da rutinski razgovara sroditeljima svojih bolesnika o sigurnom spavanju. Pritom svi ispitani PPZZ-i savjetuju roditeljima da dojenčad stave spavati u kolijevkuu sobi s njima. Njih 72% preporuča položaj na leđima, a ostali na boku. Više od pola (52%) ispitanika ne smatra da pušenje oca iliizloženost duhanskom dimu majke u trudnoći povećava rizik od SIDS-a, a 38% ih ne smatra ni da majčino pušenje povećava rizik.Većina (76%) PPZZ-a ne zna da hranjenje formulom značajno povećava rizik od SIDS-a. Polovina ispravno smatra da je zajedničkospavanje povezano s duljim trajanjem dojenja.Zaključak: Nedostatno je znanje PPZZ-a o rizičnim čimbenicima za SIDS. S obzirom na to da većina pedijatara rutinski razgovara sroditeljima dojenčadi o sigurnom spavanju, bitno je da budu upoznati s najnovijim preporukama.Aim: To assess the knowledge about risk factors for sudden infant death syndrome (SIDS) among primary health care paediatricians (PHCP) in the Split-Dalmatia County (SDC), Dubrovnik-Neretva County (DNC) and Bjelovar-Bilogora County (BBC). Subjects and methods: This cross-sectional study was conducted from February to September 2018 and included PHCPs from SDC, DNC and BBC. Paediatricians were sent a questionnaire with 18 questions on their attitudes about sleep and breastfeeding communicated to parents of their patients, and on their knowledge about SIDS. Results: The analysis included 29 questionnaires received from PHCPs, most of them aged >55. The majority of paediatricians (72%) reported they routinely talked to parents of their patients about safe sleeping. All PHCPs advised parents to put infants to sleep in a cradle in their bedroom; 72% of PHCPs advised putting infants to sleep on his/her back and the rest advised to do it on his/her side. More than half (52%) of PHCPs considered that the risk of SIDS was not increased by father’s smoking habit or exposure to maternal tobacco smoke in pregnancy, and 38% believed that neither mother’s smoking habit increased that risk. The majority (76%) of PHCPs did not know that formula feeding significantly increased the risk of SIDS. Half of the study paediatricians correctly considered mother-infant bed-sharing to be associated with longer breastfeeding. Conclusion: Study results revealed the knowledge of PHCPs about risk factors for SIDS to be inadequate. As the majority of paediatricians regularly talked to infant parents about safe sleeping, it is of utmost importance for them to be informed on the latest recommendations in the field

Microglial- and Astrocyte-Specific Expression of Purinergic Signaling Components and Inflammatory Mediators in the Rat Hippocampus During Trimethyltin-Induced Neurodegeneration

The present study examined the involvement of purinergic signaling components in the rat model of hippocampal degeneration induced by trimethyltin (TMT) intoxication (8 mg/kg, single intraperitoneal injection), which results in behavioral and neurological dysfunction similar to neurodegenerative disorders. We investigated spatial and temporal patterns of ecto-nucleoside triphosphate diphosphohydrolase 1 (NTPDase1/CD39) and ecto-5′ nucleotidase (eN/CD73) activity, their cell-specific localization, and analyzed gene expression pattern and/or cellular localization of purinoreceptors and proinflammatory mediators associated with reactive glial cells. Our study demonstrated that all Iba1+ cells at the injured area, irrespective of their morphology, upregulated NTPDase1/CD39, while induction of eN/CD73 has been observed at amoeboid Iba1+ cells localized within the hippocampal neuronal layers with pronounced cell death. Marked induction of P2Y12R, P2Y6R, and P2X4-messenger RNA at the early stage of TMT-induced neurodegeneration might reflect the functional properties, migration, and chemotaxis of microglia, while induction of P2X7R at amoeboid cells probably modulates their phagocytic role. Reactive astrocytes expressed adenosine A1, A2A, and P2Y1 receptors, revealed induction of complement component C3, inducible nitric oxide synthase, nuclear factor-kB, and proinflammatory cytokines at the late stage of TMT-induced neurodegeneration. An increased set of purinergic system components on activated microglia (NTPDase1/CD39, eN/CD73, and P2X7) and astrocytes (A1R, A2AR, and P2Y1), and loss of homeostatic glial and neuronal purinergic pathways (P2Y12 and A1R) may shift purinergic signaling balance toward excitotoxicity and inflammation, thus favoring progression of pathological events. These findings may contribute to a better understanding of the involvement of purinergic signaling components in the progression of neurodegenerative disorders that could be target molecules for the development of novel therapies

Ecto-5'-nucleotidase marks amoeboid microglial cells in the rat model of neurodegeneration

Adenosine 5'-triphosphate (ATP) and adenosine are versatile signaling molecules involved in many pathophysiological processes in the nervous system. They can be released from all types of brain cells in the extracellular space and activates purinergic receptors. Signaling via extracellular ATP is regulated by cell-surface located ectonucleotidases. Extracellular AMP resulting from the hydrolysis of ATP and ADP can in turn be hydrolyzed into adenosine by ecto-5'-nucleotidase (eN). We examined the involvement of purinergic signaling components in the rat model of trimethyltin (TMT)-induced hippocampal neurodegeneration (8mg/kg, single ip), which results in behavioral and neurological dysfunction similar as in Alzheimer's disease models. Enzyme histochemistry and immunohistochemistry (ir) showed that products of AMPase activity and eN-ir were accumulated in the neuronal strata, infiltrating within neuronal cell layers, depicting individual round-shaped elements that covered neuronal layers with pronounced cell death mostly at the late stage of TMT-induced neurodegeneration. Co-localization with Iba1+ specifically marked eN at amoeboid microglial cells. Neither of the tested pro-inflammatory cytokines (IL-1β, TNF-α, IL10) and C3 nor polarization marker iNOS was found in association with those Iba1/eN+ -cells. Iba1-ir cells co-localized with Arg1-ir and phagocytic marker CD68- ir. Marked induction of P2Y12R-, P2Y6R-, and P2X4-mRNA at the early stage of TMT-induced neurodegeneration might reflect the migration, and chemotaxis of microglia, while induction of P2X7R at amoeboid cells probably modulates their phagocytic role. These findings may contribute to a better understanding of the involvement of purinergic signaling components in the progression of neurodegenerative disorders that could be target molecules for development of novel therapies.Poster Session: Neuroimmunoendocrine Interaction

Estradiol receptors mediate estradiol-induced inhibition of mitochondrial Ca2+ efflux in rat caudate nucleus and brain stem

Our earlier studies found that in vitro estradiol modulates mitochondrial Ca2+ transport in discrete brain regions. The present study examined the role of estradiol receptors (ERs) in estradiol-induced inhibition of Ca2+ efflux from synaptosomal mitochondria isolated from rat caudate nuclei and brain stems. Radioactively labeled CaCl2 (0.6-0.75 mu Ci (CaCl2)-Ca-45) was used for Ca2+ transport monitoring. The results revealed that in the presence of ER antagonist 7 alpha, 17 beta-[9[(4,4,5,5,5-pentafluoropentyl) sulfinyl] nonyl] estra-1,3,5( 10)-triene-3,17-diol (ICI 182,780) (1 mu mol/L), the inhibitory effect of estradiol on mitochondrial Ca2+ efflux was more than 60% decreased, suggesting the involvement of ER in this mode of estradiol neuromodulatory action. When particular contributions of ER alpha and ER beta were tested, it was found that ER beta agonist 2,3-bis(4-hydroxy phenyl)-propionitrile (10 nmol/L) inhibited Ca2+ efflux more than 20%, while the inhibition with ER alpha agonist 4,4', 4''-(4-propyl-[1H]-pyrazole-1,3,5-triyl) trisphenol (10 nmol/L) was about 10%, both compared to the control. Both agonists demonstrated attenuation of Ca2+ efflux decrease in the presence of mitochondrial Na+/Ca2+ exchanger antagonist 7-chloro-5-(2-chlorophenyl)-1,5-dihyhdro-4,1-benzothiazepin-2(3H)-one (10 mu mol/L), showing interference with the inhibitory action of that agent. Our results strongly indicate ERs as the mediators of estradiol-induced mitochondrial Ca2+ efflux inhibition in rat caudate nucleus and brain stem synaptosomes

Intermittent Theta Burst Stimulation Ameliorates Cognitive Deficit and Attenuates Neuroinflammation via PI3K/Akt/mTOR Signaling Pathway in Alzheimer’s-Like Disease Model

Neurodegeneration implies progressive neuronal loss and neuroinflammation further contributing to pathology progression. It is a feature of many neurological disorders, most common being Alzheimer’s disease (AD). Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive stimulation which modulates excitability of stimulated brain areas through magnetic pulses. Numerous studies indicated beneficial effect of rTMS in several neurological diseases, including AD, however, exact mechanism are yet to be elucidated. We aimed to evaluate the effect of intermittent theta burst stimulation (iTBS), an rTMS paradigm, on behavioral, neurochemical and molecular level in trimethyltin (TMT)-induced Alzheimer’s-like disease model. TMT acts as a neurotoxic agent targeting hippocampus causing cognitive impairment and neuroinflammation, replicating behavioral and molecular aspects of AD. Male Wistar rats were divided into four experimental groups–controls, rats subjected to a single dose of TMT (8 mg/kg), TMT rats subjected to iTBS two times per day for 15 days and TMT sham group. After 3 weeks, we examined exploratory behavior and memory, histopathological and changes on molecular level. TMT-treated rats exhibited severe and cognitive deficit. iTBS-treated animals showed improved cognition. iTBS reduced TMT-induced inflammation and increased anti-inflammatory molecules. We examined PI3K/Akt/mTOR signaling pathway which is involved in regulation of apoptosis, cell growth and learning and memory. We found significant downregulation of phosphorylated forms of Akt and mTOR in TMT-intoxicated animals, which were reverted following iTBS stimulation. Application of iTBS produces beneficial effects on cognition in of rats with TMT-induced hippocampal neurodegeneration and that effect could be mediated via PI3K/Akt/mTOR signaling pathway, which could candidate this protocol as a potential therapeutic approach in neurodegenerative diseases such as AD

Time-Related Sex Diffrences in Cerebral Hypoperfusion-Induced Brain Injury

Although the model of cerebral hypoperfusion in rats has been a matter of many investigations over the years, the exact intracellular and biochemical mechanisms that lead to neuron loss and memory decline have not been clearly identified. In the current study, we examined whether cerebral hypoperfusion causes changes in hippocampal protein expression of apoptotic markers in the synaptosomal fraction and neurodegeneration in a time-dependent and sex-specific manner. Adult male and female Wistar rats were divided into two main groups, controls that underwent sham operation, and animals subjected to permanent bilateral occlusion of common carotid arteries. Both male and female rats were killed 3, 7 or 90 days following the insult. The obtained results indicate that the peak of processes that lead to apoptosis occured on postoperative day 7 and that they were more prominent in males, indicating that neuroprotective effects of certain substances (planned for future experiments), should be tested at this time point

Dietary supplementation with flaxseed oil ameliorates trimethyltin (TMT)-induced neurodegeneration and gliosis in female Wistar rats

It is increasingly apparent that the prevention/treatment of neurodegenerative disorders is not only achieved through pharmacological therapy but also through the consumption of natural products. Flaxseed oil (or linseed oil, FSO) derived from the seeds of the flax (Linum usitatissimum L.) gained worldwide awareness as a neuroprotective agent due to its high content of omega-3 polyunsaturated fatty acids (n-3 PUFAs). Thus, the aim of this study was to examine the preventive effects of dietary FSO in trimethyltin (TMT) - induced hippocampal neurodegeneration and gliosis in female Wistar rats. Animals were continuously treated with FSO (1 ml/kg, orally) for two weeks, then received a single dose of TMT (8 mg/kg, i.p.), and application of FSO continued for twenty-one days. Data have convincingly shown that FSO continuous treatment ameliorated TMT-induced neuronal loss in the CA3 hippocampal region and ameliorated astrogliosis and microgliosis. FSO treatment elevated all tested n-3 fatty acids in the hippocampus: α-linolenic acid (ALA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA), and consequently increased total amount of n-3 PUFA. However, no changes in n-6 fatty acids due to FSO treatment were observed. Consequently, FSO lowered n-6/n-3 ratio compared to TMT, having a protective effect on fatty acid profile in hippocampus. These findings support beneficial neuroprotective properties of FSO against TMT-induced model of neurodegeneration and hint at a promising preventive use of FSO in hippocampal degeneration and dysfunctionPoster Session: Brain Metabolism & Dietary Intervention