Paulina Rachwalska

Interleukin-1 � (IL-1�) level is modulated during multiple stress reactions both in brain structures involved in hypothalamicpituitary-adrenal (HPA) axis regulation and peripheral systems. Multiple distinct stressors induce different IL-1 � and HPA axis responses. The purpose of the present study was to determine if the effect of prior repeated restraint stress on IL-1 � levels in prefrontal cortex, hippocampus, hypothalamus and plasma may have an impact on alterations induced in HPA axis responses. Experiments were performed on male Wistar rats which were exposed to 10 min restraint stress twice a day for 3 days. Twenty four hours after the last stress period rats were restrained for 10 min and decapitated at 0, 1, 2 or 3 h after cessation of stress. Control rats were injected ip with saline and some of experimental groups with IL-1 � receptor antagonist (IL-1ra). After rapid decapitation, trunk blood was collected and prefrontal cortex, hippocampus and hypothalamus were excised an...

Drug delivery to the eye remains a real challenge due to the presence of ocular anatomical barriers and physiological protective mechanisms. The lack of effective siRNA delivery mechanism has hampered the real potential of RNAi therapy, but recent literature suggests that nanocarrier systems show great promise in enhancing siRNA bioavailability and reducing the need for repeated intraocular injections. A diverse range of materials are under exploration worldwide, including natural and synthetic polymers, liposomes, peptides, and dendrimeric nanomaterials. This chapter describes a simple workflow for feasibility assessment of a proposed ocular surface siRNA delivery system. Gel retardation assay is used for investigation of optimal siRNA to carrier loading ratio. Fluorescent siRNA allows for initial in vitro testing of cellular uptake to corneal epithelial cells and investigation of in vivo siRNA delivery into mouse cornea by live animal imaging and fluorescence microscopy.

BACKGROUND The aim of this study was to compare the expression of interleukin-1β (IL-1β), neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) in the prefrontal cortex (PFC), hippocampus (HIP) and hypothalamus (HT) during chronic crowding (CS) (psychosocial) and restraint (RS) (physico-psychological) stress. Adaptational changes of these stress mediators to a subsequent acute RS, in two models of chronic stress were investigated. METHODS Rats were crowded (24 in one cage) or restrained in metal tubes for 10min twice a day for 3, 7, and 14 consecutive days and decapitated. For determination of adaptational changes the chronically crowded and restrained rats 24h after the last stress session were subjected to a single 10min RS. The IL-1β, nNOS and iNOS protein levels in brain structures samples were analyzed by Western blot procedure. RESULTS Chronic CS for 3days did not markedly change the subsequent acute stress induced expression of nNOS, iNOS and IL-1β protein level in PFC and iNOS protein level in HT. CS markedly decreased the expression of nNOS, iNOS and IL-1β in HIP. By contrast, parallel chronic RS, significantly increased the subsequent acute stress-induced expression of iNOS and IL-1β in PFC and considerably increased iNOS level in HT. CONCLUSION Chronic psychosocial stress, may protect against possible harmful action of hyperproduction of iNOS and iNOS derived nitric oxide (NO) mainly in PFC and HIP. By contrast, chronic physico-psychosocial stress may strongly potentiate additional stress-induced harmful effects of NOS and IL-1β hyperproduction.

Stress response involves activation of hypothalamic-pituitary-adrenal axis and expression of nitric oxide synthase (NOS) and cyclooxygenase (COX) in different brain regions. This study evaluated whether alterations in hypothalamic COX and NOS isoenzymes levels in response to acute restraint are affected by vagotomy and prior crowding stress. Subdiaphragmatic vagotomy or sham surgery was performed 10 days before experimental procedure. Adult male Wistar rats were subjected to crowding stress for 3 days, then exposed to 10 min restraint and decapitated immediately after stress or 1, 2 and 3 hours later. Western blot analyses were performed to determine the level of COX-1, COX-2, nNOS and iNOS in hypothalamus. Acute restraint significantly decreased hypothalamic COX-1 in sham-operated rats but increased COX-1 parallel to decreasing of COX-2 in vagotomized animals. After vagotomy basal nNOS level in hypothalamus was increased compared with sham-operated rats, but nNOS expression was ins...

Aims The study evaluated whether the vagus nerve is involved in restraint-induced activation of hypothalamic-pituitary-adrenal (HPA) axis and investigated levels of constitutive and inducible cyclooxygenase (COX-1 and COX-2) in frontal cortex, hippocampus and hypothalamus in response to acute restraint stress after vagotomy. Methods Subdiaphragmatic vagotomy or sham surgery was performed 10 days before experimental procedure. Adult male Wistar rats were subjected to 10 min restraint and decapitated immediatelly after stress or 1, 2 and 3 hours later. Trunk blood for ACTH and corticosterone assays was collected and brain structures were excised for COX-1 and COX-2 levels determination by Western blot. Results Acute restraint induced a significant and relatively identical parallel increase in ACTH and corticosterone secretion immediately after stress termination. These changes disappeared an hour later and returned to control level in sham-operated rats but corticosterone remained sli...

Streszczenie Wino jest produktem złożonych interakcji mikrobiologicznych i biochemicznych między różnymi gatunkami drożdży, bakterii i grzybów strzępkowych, dlatego niezmiernie ważne jest kontrolowanie jego potencjalnego mikrobiologicznego skażenia. Szybkie metody wykrywania i identyfikacji drobnoustrojów w winach mogą umożliwić rozpoznanie wady i uratować trunek przed zepsuciem na różnych etapach jego wytwarzania. Celem pracy była izolacja i identyfikacja mikroorganizmów z psujących się win metodami klasycznymi oraz zbadanie wiarygodności tych metod za pomocą elektroforezy SDS-PAGE. Z prób zepsutych win wyizolowano 17 szczepów drożdży, które analizowano metodami klasycznymi (m.in. analiza mikroskopowa, zdolność do zarodnikowania, testy fermentacyjne i asymilacyjne, i in.). Badane szczepy wykazały bardzo zbliżone cechy, ale dopiero za pomocą rozdziału całkowitego rozpuszczalnego białka komórkowego metodą SDS-PAGE udowodniono, że wszystkie należały do gatunku Candida parapsilosis. Wy...

ABSTRACT In the present study, we examined whether the vagus nerve is involved in mediating the stimulation of hypothalamic-pituitary-adrenal (HPA) axis by acute restraint stress and whether it is affected by chronic crowding stress. Subdiaphragmatic vagotomy or sham surgery was performed 10 days before experimental procedure. Adult male Wistar rats were subjected to crowding stress for 3 days, then exposed to 10 min restraint in metal tubes and decapitated immediatelly after restraint or 1 hour later. Trunk blood was collected and adrenocorticotropic hormone (ACTH) and corticosterone levels were determined in plasma using commercially available kits. Basal plasma corticosterone level in sham operated and subdiaphragmatically vagotomized rats was increased in comparison to control non-operated rats. Three-day crowding stress alone substantially augmented the resting plasma corticosterone level in control non-operated rats. This effect was absent in sham operated and vagotomized animals. Acute restraint stress induced significant and relatively identical parallel increase in ACTH and corticosterone secretion immediately after stress termination. These changes completely disappeared an hour later. Restraint-induced ACTH and corticosterone increased levels in vagotomized and sham operated rats were lower than in non-operated animals. Crowding stress for 3 days substantially decreased the restraint-induced ACTH and corticosterone response in non-operated and sham operated rats. By contrast, prior crowding stress moderately intensified the restraint-induced ACTH and corticosterone secretion immediatelly after restraint in subdiaphragmatically vagotomized rats. These results suggest that 3-day crowding stress desensitized HPA axis response to acute restraint, possibly by the involvement of vagal afferents. This research was supported by the project Interdisciplinary PhD Studies "Molecular sciences for medicine" (co-financed by the European Social Fund).