Desipramine administered chronically inhibits lipopolysaccharide-stimulated production of IL-1ß in the brain and plasma of rats.

Nowadays, it is assumed that therapeutic efficacy of antidepressants depends, at least partly, on their anti-inflammatory properties. The present study investigated for the first time the effect of 21-day oral administration of desipramine on the lipopolysaccharide (LPS)-stimulated IL-1ß concentration in the olfactory bulb, hypothalamus, frontal cortex, hippocampus and plasma of rats, and on the LPS-induced IL-1ß mRNA level in the olfactory bulb. Desipramine (15mg/kg/day) reduced significantly the LPS (250 µg/kg i.p.)-induced IL-1ß concentration in the olfactory bulb, hypothalamus and in plasma, and diminished the LPS effect on IL-1ß mRNA in the olfactory bulb. Plasma concentration of desipramine was comparable to its therapeutic range. By using the α1/α2-adrenoceptor antagonist prazosin and the unspecific ß-adrenoceptor antagonist propranolol given prior to LPS, we found that the effect of desipramine on LPS-induced IL-1ß production was partially mediated by both adrenoceptors in the olfactory bulb and plasma, and that ß-adrenoceptors contributed also to its effect on the stimulated IL-1ß concentration in the hypothalamus. The effect of LPS on the cerebral IL-1ß levels was, in part, mediated by ß-adrenoceptors and, in a region-specific manner, by α1/α2-adrenoceptors. The findings provide evidence for central and peripheral anti-inflammatory activity of desipramine and confirm the impact of the noradrenergic system on IL-1ß production induced by an immunostimulatory challenge.

Affect of antidepressants on the in vitro differentiation of rat bone marrow mesenchymal stem cells into neuronal cells.

BACKGROUND: Bone marrow is a valuable source of mesenchymal stem cells (MSCs) that can be used in regenerative medicine. MSCs are able to differentiate into cells from all three germ layers under specific conditions. The aim of the current work was to study the differentiation of rat MSCs (rMSCs) into neuron-like cells. NEW METHOD: We investigated how the antidepressants imipramine, desipramine, fluoxetine and tianeptine affect the differentiation of rMSCs. Furthermore, we present differentiation cocktails using a cortex astrocyte-conditioned medium (CACM) separately or in conjunction with each of the antidepressants and investigated their additive effect on the efficiency of differentiation. We also observed how various differentiation conditions affect the number of primary dendrites and branching dendrites per cell. RESULTS: Gene expression for an early neuronal marker (ß-III-tubulin) and markers that are typical for adult neurons such as Th, Htr2A and Slc6a4 were observed. The Tubb3 and Htr2A gene expression were up-regulated, Th decreased slightly and Slc6a4 was down-regulated after differentiation We observed a two-fold higher percentage of ß-III-tubulin positive cells after treatment with antidepressants and two-fold increase of neuron-like cells after using CACM with imipramine or fluoxetine simultaneously. Differentiation using imipramine or in conjunction with CACM and desipramine or fluoxetine simultaneously increased the number of cell dendrites. COMPARISON WITH EXISTING METHODS: The results that were obtained are completely new and need further investigations in the nearest future. CONCLUSIONS: These results suggest that antidepressants improve differentiation efficiency of rMSCs and may be useful in the preparation of rMSCs for transplantation. Differentiation efficiency is higher after long-term exposure to antidepressants, than after a 24-h exposure. Nearly additive effect of CACM and imipramine or fluoxetine suggests a beneficial role of antidepressants after transplantation.

Chronic social instability stress enhances vulnerability of BDNF response to LPS in the limbic structures of female rats: a protective role of antidepressants.

The aim of the present study was to estimate the influence of antidepressants given chronically on brain-derived neurotrophic factor (BDNF) alterations induced by lipopolysaccharide (LPS) in the amygdala and hippocampus of female rats subjected to chronic social instability stress (CSIS) for 29-30 days. CSIS was used as a paradigm known to be more stressful for females because stress induces affective disorders more frequently in women than men. An increased relative adrenal weight and a tendency towards the enhanced plasma corticosterone concentration were found in the stressed rats. Sucrose preference was not changed. On the last experimental day, the rats in the estrus phase were injected ip with LPS (1mg/kg). In the stressed rats, LPS administration decreased BDNF mRNA levels in both limbic structures. Desipramine (10mg/kg), fluoxetine (5mg/kg) or tianeptine (10mg/kg) given ip once daily reversed the effect of the combined stress and LPS, and tianeptine induced the strongest effects. These results indicate that chronic stress enhances vulnerability of BDNF response to deleterious influence of neuroinflammation in the examined limbic structures, what may account for its role in triggering neuropsychiatric diseases. The observed effect of antidepressants may be of significance for their therapeutic effects in the stress-induced affective disorders in females.

Biological agents database in the armed forces.

Rapid detection and identification of the biological agent during both, natural or deliberate outbreak is crucial for implementation of appropriate control measures and procedures in order to mitigate the spread of disease. Determination of pathogen etiology may not only support epidemiological investigation and safety of human beings, but also enhance forensic efforts in pathogen tracing, collection of evidences and correct inference. The article presents objectives of the Biological Agents Database, which was developed for the purpose of the Ministry of National Defense of the Republic of Poland under the European Defence Agency frame. The Biological Agents Database is an electronic catalogue of genetic markers of highly dangerous pathogens and biological agents of weapon of mass destruction concern, which provides full identification of biological threats emerging in Poland and in locations of activity of Polish troops. The Biological Agents Database is a supportive tool used for tracing biological agents' origin as well as rapid identification of agent causing the disease of unknown etiology. It also provides support in diagnosis, analysis, response and exchange of information between institutions that use information contained in it. Therefore, it can be used not only for military purposes, but also in a civilian environment.

Imipramine and fluoxetine inhibit LPS-induced activation and affect morphology of microglial cells in the rat glial culture.

BACKGROUND: Recent evidence has suggested that antidepressants evoke neuroprotective and immunomodulatory effects in the brain, partly at least, by inhibiting glia activation. This study has been conducted on the lipopolysaccharide (LPS)-stimulated primary rat mixed glial cell culture in order to better recognize the influence of imipramine (a tricyclic antidepressant) and fluoxetine (a selective serotonin reuptake inhibitor) on the important balance between pro- and anti-inflammatory cytokines produced by the glial cells. Moreover, microscopic observations were made to describe the morphological alterations in the studied cell cultures exposed to the drugs. METHODS: The effect of both antidepressants on TNF-α, IL-1ß and IL-10 levels was determined by ELISA. The mRNA levels of mentioned cytokines were evaluated by qRT-PCR assay. Moreover, drug influence on the LPS-stimulated level of NF-κB p65 subunit in nuclear fraction was determined by the colorimetric transcription factor assay. RESULTS: After LPS-stimulation both drugs decreased concentration of TNF-α and IL-1ß in culture medium and expression of TNF-α and IL-1ß mRNAs in cellular extracts. They also diminished the LPS-induced nuclear translocation of NF-κB p65 subunit. In contrast, imipramine and fluoxetine induced a few-fold weaker suppressing effect on the levels of IL-10. Parallelly to the inhibition of the LPS-induced inflammatory response, the antidepressants prevented the morphological alterations of cells elicited by LPS. Moreover, in unstimulated cultures imipramine but not fluoxetine caused transformation of microglia cells into cells with neuron-like morphology. CONCLUSIONS: Imipramine and fluoxetine, by modulating glia activation, may exert anti-inflammatory effects in the CNS. It also seems that microglia cells are important target particularly for imipramine.

State-of-the-art in biosafety and biosecurity in European countries.

The terms biosafety and biosecurity are widely used in different concepts and refer not only to protection of human beings and their surrounding environment against hazardous biological agent, but also to global disarmament of weapons of mass destruction. As a result, the biosafety and biosecurity issues should be considered interdisciplinary based on multilateral agreements against proliferation of biological weapons, public health and environmental protection. This publication presents information on both, international and national biosafety and biosecurity legislation. Status of national implementation of the Biological and Toxin Weapons Convention, penalization issues and measures to account for and secure production, use, storage of particularly dangerous pathogens or activities involving humans, plants and animals where infection may pose a risk have been analyzed. Safety and security measures in laboratories have been studied. Moreover, dual-use technology and measures of secure transport of biohazard materials have been also taken into account. In addition, genetic engineering regulations, biosecurity activities in laboratories and code of conducts have been investigated, as well.

Severe influenza outbreak in Western Ukraine in 2009--a molecular-epidemiological study.

INTRODUCTION: In the autumn of 2009 the authors participated in a humanitarian operation in Western Ukraine by undertaking an epidemiological investigation of an influenza-like-illness (ILI) in the L'viv Oblast region. Mobile biological survey teams took samples from civilian patients with severe acute respiratory distress syndrome, rapid transportation of the samples, and their molecular analysis in Poland to provide accurate results. OBJECTIVE: The aim of the study was the molecular and epidemiological analysis of the biological samples collected. MATERIAL AND METHODS: Real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR), multiplex PCR techniques, traditional Sanger Sequencing and classical viral culture methods were used. RESULTS: Among the 124 influenza-like illness cases, ~50% (58) were positive for influenza A virus in WHO-CDC molecular assay, including subtyping. The specimens were further analyzed to confirm results and determine the genetic sequence. Phylogenetically, the nucleotide similarity of both the Ukraine specimens and reference A/California/7/2009 (pH1N1) was 99.2-99.3%. Oseltamivir resistance was not registered. HA1 region characterization showed an overall protein identity of 98.5-99.4%. CONCLUSIONS: An unexpected high contribution of influenza A was confirmed among ILI patients, as well as a very limited number of other detected viruses, indicate that the 2009 epidemic in western Ukraine was strongly related to novel influenza A/H1N1. The importance of swift sharing of information and reference laboratories networking in surveillance, as well as serving governments and international agencies in pursuing adequate actions, should be stressed.

Antidepressant drugs as a complementary therapeutic strategy in cancer.

In the last decade, it has been increasingly recognized that antidepressant drugs may exert a range of effects, in addition to their well-documented ability to modulate neurotransmission. Although as a group they act on monoaminergic systems and receptors in different ways, a number of studies have demonstrated that at least some antidepressants might have other properties in common, including immunomodulatory, cyto/neuroprotective, analgesic and anti-inflammatory activities. These properties are partly related to the influence of antidepressants on glial cell function. Recently, emerging information about the possible anticancer properties of antidepressants has sparked increased interest within scientific community, and there is now evidence that these drugs affect the key cellular mechanisms of carcinogenesis. This review examines the putative cellular targets for the anticancer action of antidepressant drugs, and presents examples of the interaction between antidepressants and anticancer drugs. By reviewing the current state of research in this area, we hope to focus the attention of oncologists and researchers engaged in the study of cancer on the role that antidepressant drugs could play in the complementary therapy of cancer.

[Analysis of the prevalence of HBV markers relevant to the risk of reactivation of infection in patients with hematologic diseases]. / Analiza wystepowania markerów wirusa HBV istotnych dla ryzyka reaktywacji zakazenia u osób z chorobami hematologicznymi.

The study assessed the incidence of HBV markers (HBsAg, anti-HBc, anti-HBs) important for determination of the risk of reactivation of infection, with particular interest of occult infection (presence of HBV DNA in the absence of HBsAg) in patients treated at the Institute of Hematology and Transfusion Medicine. Anti-HBc frequency was correlated with the age and sex of patients. HBsAg was detected in 16/468 examined patients, 98/468 (21%) were anti-HBc positive. HBV DNA was detected in 41/98 anti-HBc positives; in 13 simultaneously with HBsAg. 28 patients had occult HBV infection (HBV DNA+/HBsAg). Antibody to HBsAg was detected in 163/430 (38%) patients, 81 out of them on protective level (> 100 IU/l). It was shown that occult HBV infection occurs in approximately 6% of patients. In most of them the protective levels of anti-HBs are detected.

Atorvastatin and fenofibric acid differentially affect the release of adipokines in the visceral and subcutaneous cultures of adipocytes that were obtained from patients with and without mixed dyslipidemia.

In this study, we compared the effects of atorvastatin and fenofibric acid, which were administered alone or in combination, on the secretory function of human adipocytes that were obtained from the visceral and subcutaneous adipose tissues of 19 mixed dyslipidemic patients and 19 subjects with a normal lipid profile. The adipocytes were incubated in vitro in the presence of atorvastatin and/or fenofibric acid. The secretory function of the cells was determined using ELISA assays. The visceral adipocytes released significantly more adiponectin and IL-6 and less PAI-1 than those that were obtained from subcutaneous tissue. The levels and patterns of adipokine release differed between the patients with or without lipid abnormalities and between the adipocytes that were obtained from visceral or subcutaneous adipose tissue. The culture that contained hypolipidemic drugs resulted in the significant changes of the release of adipokines. The effects of atorvastatin and fenofibric acid on the hormonal function of human adipocytes may be, in part, responsible for the clinical efficacy of these drugs in the prevention and treatment of dyslipidemia-related cardiovascular and metabolic disorders. The study supports the concept that the pleiotropic effects of fenofibrate and atorvastatin may be, in part, a result of their impact on the secretory function of adipocytes.

Immunosuppressant cytoprotection correlates with HMGB1 suppression in primary astrocyte cultures exposed to combined oxygen-glucose deprivation.

The protective potential of immunosuppressants has been reported in many experimental models of ischemia both in vivo and in vitro, suggesting a novel therapeutic application of these drugs. Because high-mobility group box 1 (HMGB1) protein has recently been reported to be involved in ischemic brain injury, the purpose of the present study was to determine whether treatment with immunosuppressants could decrease the expression and release of HMGB1 in astrocytes exposed to simulated ischemic conditions (combined oxygen-glucose deprivation, OGD). We also investigated whether immunosuppressive drugs could attenuate necrosis in astrocyte cultures exposed to OGD. Finally, we studied the influence of immunosuppressants on the expression of NFκB, inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). Cells were treated with cyclosporine A, FK506 and rapamycin (all drugs at concentrations of 0.1, 1 and 10 µM). Our study provides evidence that immunosuppressants decrease the expression and release of HMGB1 in ischemic astrocytes. Our data suggest that HMGB1 release may be partly an active process triggered by oxidative stress because the antioxidant N-acetylcysteine (NAC) clearly attenuated HMGB1 expression and release. Furthermore, we show that the immunosuppressants, at the same concentrations that significantly suppressed HMGB1 expression and release, were also able to prevent the necrosis of ischemic astrocytes and inhibit the expression of inflammatory mediators (NFκ, iNOS and COX-2). These results provide further information about the cytoprotective mechanisms of immunosuppressants on ischemic astrocytes, especially in relation to the pathophysiology of ischemic brain injury. It appears that the protective effects of immunosuppressants can be mediated in part by the suppressing the expression and release of HMGB1 in astrocytes, which leads to the attenuation of ischemia-induced necrosis and neuroinflammation.

[Cultivated oral epithelial cells in oral cavity tissue losses treatment with autologous transplantation--case reports]. / Leczenie ubytków tkanek w jamie ustnej przy pomocy autologicznego przeszczepu nablonka uzyskanego droga hodowli tkankowej--opis przypadków.

UNLABELLED: The rapid development of bioengineering in recent years enables to search for new therapies involving the use of tissue cultures. The aim of this paper is to study and apply the method of keratinocytes culture of mucous membrane on amniotic membrane to cover the losses in the oral cavity. MATERIAL AND METHODS: In four cases keratinocytes culture on amniotic membrane was applied and transplanted to cover antro-oral fistulas and mucosal and mandibular bone losses. RESULTS: Histopathological and immunohistochemistrical examinations of the culture were performed. Cytokeratin 14, protein 63, conexin 43 and IgG were indicated. CONCLUSION: Autologous transplants of epithelial cells on amniotic membrane are a new effective method to cover unhealed tissue losses in oral cavity with the use of modern methods of tissue engineering.

Culture morphology of the autologous cultivated corneal epithelium.

PURPOSE: Ocular surgery based on cultivated corneal epithelium has become a very promising procedure eligible to restore the ocular surface. Analysis of morphologic features and the phenotype of cultivated epithelial cells determines their quality and eligibility of transplantation. MATERIAL AND METHODS: Corneal epithelial cultures were carried out in 25 patients suffering from limbal deficiency after chemical or thermal burns. Fellow healthy eyes were the source of limbal epithelium for the culture. Limbal cells from a 2 mm2 biopsy were seeded on an amniotic membrane after enzymatic pretreatment. Cultures were carried in standard conditions in a supplemented DMEM HAM/F12 medium in the presence of 3T3 fibroblasts. Light microscopy was used to analyze the regularity of the cultivated epithelial layer, histologic examination was used to establish number of epithelial layers, and immunohistochemistry for epithelial and proliferation markers was applied to confirm cell origin and proliferative potential. Staining for cytokeratin 3, 12, 19, connexin 43, and protein p63 was performed. RESULTS: In 25 donors, 27 cultures of the epithelium were performed. In 2 cases, plates were contaminated. Both cultures were repeated. In 84% of the cultures, regular stratified growth of the epithelium with complete covering of amniotic membrane was observed. In 16% of cultures, growth was not regular, showing differences in the number of cell layers. Staining for cytokeratin 3/12 confirmed the corneal origin of cultivated epithelia. The number of epithelial layers ranged from 3 to 9; the average was 5.3 +/- 1.9 layers. CONCLUSION: Cultures of limbal epithelial cells are a valuable source of tissue for restoration of the corneal epithelium.

Cytosolic phospholipase A2 inhibition is involved in the protective effect of nortriptyline in primary astrocyte cultures exposed to combined oxygen-glucose deprivation.

The protective potential of nortriptyline has been reported in a few experimental models of brain ischemia, both in vivo and in vitro. However, the detailed molecular mechanisms of the protective action of the drug are still unresolved. The aim of the present study was to determine whether treatment with low or medium concentrations of nortriptyline (0.1-10 µM) might have an effect on cPLA2 protein and/or mRNA expression in ischemic astrocytes, and whether this influence might be related to its potential positive influence on cell viability. On the 21(st) day in vitro, primary cultures of rat cortical astrocytes were subjected to ischemia-simulating conditions (combined oxygen glucose deprivation, OGD) for 24 h and exposed to nortriptyline. The drug at concentrations of 0.1 and 1 µM attenuated the expression of cPLA2 (both the phosphorylated and unphosphorylated forms) together with a significant decrease in the cPLA2 mRNA level in ischemic astrocytes. We have demonstrated that nortriptyline influences a decrease in cPLA2-mediated arachidonic acid (AA) release through a mechanism that appears to involve the attenuation of both PKC and Erk1/2 kinase expression. Nortriptyline also significantly prevented mitochondrial depolarization in ischemic astrocytes. Moreover, the antidepressant protected glial cells against OGD-induced apoptosis and necrosis. Our findings document a role for cPLA2 expression attenuation and AA release inhibition in the protective effect of nortriptyline in ischemic astrocytes.

Quantification of metformin by the HPLC method in brain regions, cerebrospinal fluid and plasma of rats treated with lipopolysaccharide.

Recently, it has been reported that metformin may attenuate inflammation and directly act on the central nervous system. Using the HPLC method, in Wistar rats, we assessed the changes in metformin concentrations in various brain regions (pituitary gland, olfactory bulb, hypothalamus, cerebellum, hippocampus, striatum, frontal cortex), cerebrospinal fluid and plasma after single and chronic oral administration, in the model of systemic inflammation induced by lipopolysaccharide (ip). Regarding the influence of systemic inflammation on metformin distribution, the pituitary gland demonstrated the highest its level after single and chronic administration (28.8 ± 3.5 nmol/g and 24.9 ± 3.2 nmol/g, respectively). We concluded that orally-dosed metformin rapidly crosses the blood-brain barrier and differently accumulates in structures of the central nervous system.

Antiapoptotic action of lithium and valproate.

Lithium and valproate (VPA) have been the most widely prescribed mood stabilizers for the therapy of bipolar disorders (BD) for more than 50 years. However, the precise molecular mechanism of their pharmacological activity is not fully known. Recent studies have suggested that both drugs exert antiapoptotic effects. This review focuses on the influence of lithium and VPA on intracellular apoptotic signaling pathways. The active sites, which are implicated in mediating their action, have been described. It has been found that both drugs block the key proapoptotic molecules (GSK-3beta, caspase cascades) and enhance survival pathways (ERK1/2 and Bax proteins). The potential significance of the reported antiapoptotic effects has been discussed.