How Does CBG Administration Affect Sphingolipid Deposition in the Liver of Insulin-Resistant Rats?

BACKGROUND: Cannabigerol (CBG), a non-psychotropic phytocannabinoid found in Cannabis sativa plants, has been the focus of recent studies due to its potential therapeutic properties. We proposed that by focusing on sphingolipid metabolism, which plays a critical role in insulin signaling and the development of insulin resistance, CBG may provide a novel therapeutic approach for metabolic disorders, particularly insulin resistance. METHODS: In a rat model of insulin resistance induced by a high-fat, high-sucrose diet (HFHS), we aimed to elucidate the effect of intragastrically administered CBG on hepatic sphingolipid deposition and metabolism. Moreover, we also elucidated the expression of sphingolipid transporters and changes in the sphingolipid concentration in the plasma. RESULTS: The results, surprisingly, showed a lack of changes in de novo ceramide synthesis pathway enzymes and significant enhancement in the expression of enzymes involved in ceramide catabolism, which was confirmed by changes in hepatic sphingomyelin, sphinganine, sphingosine-1-phosphate, and sphinganine-1-phosphate concentrations. CONCLUSIONS: The results suggest that CBG treatment may modulate sphingolipid metabolism in the liver and plasma, potentially protecting the liver against the development of metabolic disorders such as insulin resistance.

Cell cycle-specific phosphorylation of p53 protein in A549 cells exposed to cisplatin and standardized air pollutants.

Exposure to particulate matter is associated with DNA damage and the risk of lung cancer. Protein p53 is activated by multi-site phosphorylation in the early stages of DNA damage and affects cell outcome. Our study aimed to assess the effect of (100 µg/mL-1/24 h) standardized air pollutants: carbon black (CB), urban dust (UD), and nanoparticle carbon black (NPCB) on cell cycle, DNA damage and 53 phosphorylation at Ser 9, Ser 20, Ser 46, and Ser 392 in proliferating and quiescent A549 cells and in cells that survived cisplatin (CisPT) exposure. Phosphorylated p53 was quantified in cell subpopulations by flow cytometry using specific fluorochrome-tagged monoclonal antibodies and analysis of bivariate fluorescence distribution scatterplots. CisPT, UD and NPCB increased site-specific p53 phosphorylation producing unique patterns. NPCB activated all sites irrespectively on the cell cycle, while the UD was more selective. p53 Ser 9-P and p53 Ser 20-P positively correlated with the numbers of CisPT-treated cells at G0/G1, and NPCB and NPCB + CisPT produced a similar effect. A positive correlation and integrated response were also found between Ser 20-P and Ser 392-P in resting A549 cells treated with NPCB and CisPT but not UD. Interdependence between the expression of p53 phosphorylated at Ser 20, and Ser 392 and cell cycle arrest show that posttranslational alterations are related to functional activation. Our data suggest that p53 protein phosphorylation in response to specific DNA damage is driven by multiple independent and integrated pathways to produce functional activation critical in cancer prevention and treatment.

Cytotoxicity of Methacrylate Dental Resins to Human Gingival Fibroblasts.

This study aimed to assess the acute and delayed cytotoxicity of three, popular light-cured methacrylate-based restorative resins (MRs): Charisma (C), Estelite (E), and Filtek (F), to human gingival fibroblasts in culture. Cells were grown for up to 24 h with light-cured (or pre-cured) resins. We evaluated resin cytotoxicity, redox imbalance, necrosis/apoptosis, miR-9, and heat shock protein 70 (HSP70). The role of resin-induced oxidative stress (damage) in HSP70-response (repair) was assessed using binary fluorescence labeling. All MRs decreased viable cell numbers and cell proliferation and damaged cell membranes, and their 24 h-delayed toxicity was lower (C), higher (F), or similar (E) to that induced by freshly-cured resins. Cell membrane damage induced by C and E decreased with time, while F produced a linear increase. All resins generated intracellular oxidative stress with the predominant necrotic outcome, and produced heterogeneous responses in miR-9 and HSP70. The double fluorescence (damage/repair) experiments pointed to common features of E and F but not C. In the subset of cells, the binary response induced by E and F was different from C, similar to each other, and positively interrelated. Experimental data show that selective MR cytotoxicity should be taken into account when considering repetitive use or massive reconstruction.

Cytotoxicity and Oxidative Stress Induction by Root Canal Sealers in Human Periodontal Ligament Fibroblasts: An in vitro Study.

Introduction: The aim of the study was to investigate the in vitro cytotoxicity, the profile of cell death, and the level of oxidative stress in human periodontal ligament fibroblasts (HPdLFs) after exposure to selected root canal sealers. Methods and Materials: Freshly mixed or set Endomethasone N (EN), RealSeal (RSEAL), Roeko Seal Automix (RSA), and Sealapex (SP) were incubated with HPdLFs. Fluorescein isothiocyanate (FITC)-annexin V (AnV) and propidium iodide (PI) staining followed by flow cytometry was used to identify the effects of the materials on cell viability and the profile of cell death. 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) with fluorescence-activated cell sorting was used to determine reactive oxygen species (ROS) formation in HPdLFs. Statistical analyses were performed using one-way ANOVA followed by post hoc tests, and significance was determined at P<0.05. Results: All materials reduced the viability of the cultured cells compared with the controls (P<0.05). Fresh SP and EN, and set RSA generated an increase of necrotic cells (P<0.05), whilst fresh RSEAL and RSA induced an elevation of apoptotic cells (P<0.001). Set RSEAL caused a rise in both apoptotic and necrotic cells compared with the controls (P<0.05). Fresh EN, RSEAL, and SP resulted in increased intracellular ROS generation compared with the negative control (P<0.001), whilst fresh RSA and all set materials were ineffective. Conclusions: This in vitro study showed us the materials tested were characterized by differentiated cytotoxic effects on HPdLFs. The fresh and set forms of sealers were capable of eliciting toxic action, inducing apoptosis and/or necrosis in HPdLFs. The toxic effects of fresh EN, RSEAL, and SP might have been due to the induction of oxidative stress in human periodontal fibroblasts. The cytotoxicity of RSA seemed to be related to the involvement of other mechanisms.

Pharmacological inhibition of sphingosine-1-phosphate lyase partially reverses spatial memory impairment in streptozotocin-diabetic rats.

Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid with strong neuroprotective properties that is important for normal excitability and synaptic transmission in the hippocampal neurons. Considering the above, the aim of the present study was to determine whether increasing brain S1P level is able to reverse spatial memory impairment in streptozotocin-diabetic rats. The experiment was carried out on diabetic (n = 22) and nondiabetic (n = 10) male Wistar rats. Diabetes was induced by a single injection of streptozotocin. Eleven weeks later, 11 diabetic animals received injections of THI (S1P lyase inhibitor) for seven days. During the last five days of the experiment spatial reference memory acquisition and retention were tested in the Morris water maze task. The animals were then anaesthetized and samples of the hippocampus, prefrontal cortex, striatum, and cerebellum were excised. The content of S1P and related sphingolipids was measured using a HPLC method. Diabetes induced a depletion of ceramide in the hippocampus and cerebellum that was associated with impaired spatial memory and learning. Administration of THI to the diabetic animals prevented ceramide depletion in the hippocampus and cerebellum, and induced an increase in S1P content in all examined brain structures. These effects were associated with an improvement in spatial memory. We conclude that pharmacological inhibition of S1P lyase partially reverses the impairment in spatial memory induced by chronic hyperglycemia, and that this effect may be related to the prevention of ceramide depletion in the hippocampus and cerebellum, the increase in brain S1P level, or both.

Sialylation pattern in lung epithelial cell line and Siglecs expression in monocytic THP-1 cells as cellular indicators of cigarette smoke - induced pathology in vitro.

PURPOSE: Cellular response to cigarette smoke (CS) involves activation of recognition receptors resulting in changes in immune status, oxidative stress and cell turnover. We investigated the effects of CS on sialic acid-binding immunoglobulin type lectins (Siglecs) expression and their sialylated ligands in human immune and non-immune cells. METHODS: Human monocytes (THP-1) and epithelial cells (A549) were cultured in CS-conditioned medium (CSM). Expression of Siglec-8 and Siglec-5/Siglec-14 was analysed in THP-1 cells using flow cytometry. The effects of CS on immune activity was evaluated flow cytometrically in these cells by assessment of phagocytosis and intracellular expression IL-1ß and IL-10. Detection and differentiation of sialic acids was analyzed by dot blot, western blot and flow cytometry using plant lectins and antibodies. RESULTS: Exposure to CS significantly increased expression of Siglec-8 and Siglec-5/Siglec-14 in THP-1 cells. These changes were accompanied by enhanced intracellular level of IL-1ß and IL-10 but reduced phagocytic activity. In THP-1 and A549 cells, the level of α2,3-sialic acids, but not α2,6-sialic acid, was significantly increased when compared to naïve cells. The level of α2,8-sialic acids increased significantly in A549 cells, but not in THP-1 cells, after exposure to CS. CONCLUSION: These results show that cellular response to CS involves changes in expression of Siglec receptors and sialylated ligands functionally associated with immunity.

Effect of root canal sealers on human periodontal ligament fibroblast viability: ex vivo study.

The aim of the study was to compare ex vivo the toxic effects of six root canal sealers immediately after mixing or setting on human periodontal ligament fibroblasts (HPdLF). Freshly mixed (I group) or set (allowed to dry for 24 h) (II group) specimens of AH Plus Jet (AH), Apexit Plus (AP), MTA Fillapex (FL), GuttaFlow (GF), MetaSEAL Soft (META), and Tubli-Seal (TS) were prepared. HPdLF were exposed for 24 h to the specimens. 3-(4,5-dimethylthiazolo-2-yl)-2,5-diphenyltetrazolium bromide assay was used to examine the effect of the root canal sealers on mitochondrial metabolic activity. Fluorescein isothiocyanate (FITC)-annexin V (AnV) and propidium iodide staining followed by flow cytometry was used to identify the effects of the materials on cell apoptosis/necrosis. Statistical analyses were performed by one-way ANOVA followed by post hoc tests, and significance was determined at P < 0.05. Most materials from the two groups reduced the viability of the cultured cells compared with the control group (P < 0.05). Statistical analysis showed significant differences in HPdLF viability between the individual materials in each group (P < 0.001). AH and AP induced a significant increase in the percentage of apoptotic cells, while TS, FL, and META elevated the proportion of necrotic cells compared with other materials and the controls (p < 0.05). The cytotoxic effects of the tested root canal sealers (both fresh and set) on HPdLF varied. Both forms of sealers were able to cause toxic effects by inducing apoptosis and necrosis in HPdLF. The cytotoxicity of FL, META, TS was mainly associated with necrosis, while AH and AP with apoptosis.

Evaluation of the cytotoxicity of selected conventional glass ionomer cements on human gingival fibroblasts.

BACKGROUND: Dentistry materials are the most frequently used substitutes of human tissues. Therefore, an assessment of dental filling materials should cover not only their chemical, physical, and mechanical characteristics, but also their cytotoxicity. OBJECTIVES: To compare the cytotoxic effects of 13 conventional glass ionomer cements on human gingival fibroblasts. MATERIAL AND METHODS: The assessment was conducted using the MTT test. Six samples were prepared for each material. Culture plates with cells and inserts with the materials were incubated at 37°C, 5% CO2, and 95% humidity for 24 h. Then the inserts were removed, 1 mL of MTT was added in the amount of 0.5 mg/1 mL of the medium, and the samples were incubated in the described conditions without light for 2 h. The optical density was measured with an absorption spectrophotometer at a wavelength of 560 nm. RESULTS: The cytotoxic effects of the Argion Molar was significantly stronger than the Fuji Triage (p = 0.007), Chemfil Molar (p < 0.0001), and Ionofil Molar AC Quick (p < 0.001). The Fuji IX GP and Fuji IX Extra had a significantly stronger adverse effect than the Chemfil Molar (p = 0.014, p = 0.029, respectively) and Ionofil Molar AC Quick (p = 0.017, p = 0.034, respectively). The cements from the low cytotoxicity group were significantly more toxic vs materials whose presence resulted in fibroblast growth (p < 0.001). CONCLUSIONS: The research conducted indicates that, although the materials studied may belong to the same group, they are characterized by low, yet not uniform, cytotoxicity on human gingival fibroblasts. The toxic effects should not be assigned to a relevant group of materials, but each dentistry product should be evaluated individually.

Evaluation of cytotoxicity and pH changes generated by various dental pulp capping materials - an in vitro study.

INTRODUCTION: Various materials are used in direct dental pulp capping method. Their biocompatibility and alkalizing abilities are of primary importance affecting therapeutic effects. The aim of this study was to evaluate and compare the cytotoxicity of various pulp-capping materials on human gingival fibroblasts and investigate the pH changes induced by these materials. MATERIAL AND METHODS: Human gingival fibroblasts were cultured with nine direct pulp materials using culture plate inserts. The cytotoxic effects were recorded by using an MTT-based colorimetric assay after 3 and 24 h. In the second part of the experiment, the materials were inserted in dialysis tubes and transferred into plastic vials containing deionized water. The changes of the medium pH were measured after 3 and 24 h. RESULTS: We showed differences in cell viability of gingival fibroblasts after varied time of exposition for the tested materials. Cell viability after 24 h increased for Dycal, Biopulp, and Calcipro, and decreased for Calcipulpe, Angelus, Angelus White, and ProRoot Regular. Cell viability for ProRoot and Life did not change. Non-setting calcium hydroxide preparations followed by the MTA group and setting calcium hydroxide materials produced the highest pH. All the tested materials significantly increased pH (p < 0.0001) at 24 h. CONCLUSIONS: Currently used pulp capping materials varied in their cytotoxicity relative to human gingival fibroblasts and their alkalizing capacities. Since most likely pH does not affect the viability of cultured cells, further investigations are required to determine physicochemical properties of these materials and the biological activity of the dental pulp.

Assessment of cytotoxic potential of root canal sealers after hardening - an ex vivo study.

AIM: The aim of this study was to perform a comparative assessment of the toxic action of root canal sealers currently on the market on human gingival fibroblasts after setting. MATERIAL/METHODS: The inserts with an equal quantity of set root canal sealers were transferred into 24-well culture dishes containing human gingival fibroblasts cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum (FCS). The dishes with materials were incubated at 37°C, 100% humidity and in an atmosphere of 5% CO2 for 24 h. The cytotoxic effects of the root canal materials were measured by the mitochondrial succinate dehydrogenase activity in living cells using tetrazolium bromide (MTT assay). RESULTS: Epiphany and Sealapex exhibited high toxicity towards human gingival fibroblasts - 25.57% ± 0.88 and 27.63 % ± 2.35 respectively (less than 30% live cells in the culture). The remaining materials were characterized by lack of a cytotoxic effect (over 90% of live cells in the culture). None of the preparations exhibited moderate or low toxicity. CONCLUSIONS: The majority of root canal sealers tested after hardening were well tolerated by human gingival fibroblasts. Only two materials were characterized by high toxicity: with methacrylate (Epiphany) and calcium hydroxide (Sealapex).

CYP3A4-dependent cellular response does not relate to CYP3A4-catalysed metabolites of C-1748 and C-1305 acridine antitumor agents in HepG2 cells.

High CYP3A4 expression sensitizes tumor cells to certain antitumor agents while for others it can lower their therapeutic efficacy. We have elucidated the influence of CYP3A4 overexpression on the cellular response induced by antitumor acridine derivatives, C-1305 and C-1748, in two hepatocellular carcinoma (HepG2) cell lines, Hep3A4 stably transfected with CYP3A4 isoenzyme, and HepC34 expressing empty vector. The compounds were selected considering their different chemical structures and different metabolic pathways seen earlier in human and rat liver microsomes C-1748 was transformed to several metabolites at a higher rate in Hep3A4 than in HepC34 cells. In contrast, C-1305 metabolism in Hep3A4 cells was unchanged compared to HepC34 cells, with each cell line producing a single metabolite of comparable concentration. C-1748 resulted in a progressive appearance of sub-G1 population to its high level in both cell lines. In turn, the sub-G1 fraction was dominated in CYP3A4-overexpressing cells following C-1305 exposure. Both compounds induced necrosis and to a lesser extent apoptosis, which were more pronounced in Hep3A4 than in wild-type cells. In conclusion, CYP3A4-overexpressing cells produce higher levels of C-1748 metabolites, but they do not affect the cellular responses to the drug. Conversely, cellular response was modulated following C-1305 treatment in CYP3A4-overexpressing cells, although metabolism of this drug was unaltered.

Prognostic relevance of HSP70 antigen and antibody measurement in patients with acute myeloid leukemia of intermediate and unfavorable cytogenetic risk.

INTRODUCTION: Heat shock proteins (HSPs) are overexpressed in many types of cancers and are implicated in tumor cell proliferation, differentiation, invasion, metastasis, death, and recognition by the immune system. It has been postulated that the HSP70 protein can be used as a prognostic indicator of overall patient survival in many types of cancer including leukemia. OBJECTIVES: The aim of the study was to evaluate the concentrations of anti­HSP70 antibody and its antigen in the peripheral blood of patients with acute myeloid leukemia (AML), as well as to assess the usefulness of this measurement. PATIENTS AND METHODS: The study included 80 patients with AML of intermediate and high cytogenetic risk, scheduled for allogenic stem cell transplantation after initial intensive chemotherapy. Plasma concentrations of anti­HSP70 antibodies and HSP70 antigen were measured by enzyme­linked immunosorbent assay. The antigen was additionally measured by Western blot analysis. The control group consisted of healthy subjects. RESULTS: Patients with AML had significantly higher anti­HSP70 antibody concentrations compared with the control group. The concentration of HSP70 antigen as well as anti­HSP70 antibody showed no associations with the type of response after induction chemotherapy. However, patients with higher antigen levels and lower anti­HSP70 antibody levels had significantly shorter overall survival. CONCLUSIONS: The study suggests that anti­HSP70 antibodies and HSP70 antigen may be valuable indicators of a poor prognosis in patients with AML.

Is mycobacterial heat shock protein 16 kDa, a marker of the dormant stage of Mycobacterium tuberculosis, a sarcoid antigen?

We demonstrated opposite presence of mycobacterial heat shock proteins (Mtb-hsp) 70 kDa, 65 kDa, 16 kDa in sera and lymph nodes in sarcoidosis (SA). Higher occurrence of serum Mtb-hsp70 than Mtb-hsp 65 and Mtb-hsp 16 could be caused by sequestration of Mtb-hsp 65 and Mtb-hsp 16 in circulating immune complexes (CIs). It is possible that in genetically different predisposed hosts, Mtb-hsp 16 induced by dose-dependent nitrate/nitrite (NOx) may be involved in latent tuberculosis (TB), active TB, or SA development. We evaluated Mtb-hsp 70, Mtb-hsp 65, Mtb-hsp 16 presence in precipitated CIs and serum NOx level in 20 SA patients, 19 TB patients, and 21 healthy volunteers using PEG precipitation, Western Blot, and Griess methods. We revealed higher NOx concentrations in SA and TB than in controls, but lower in SA than TB. Mtb-hsp 16, Mtb-hsp 65, and Mtb-hsp70 concentrations in precipitated CIs were higher in SA than in TB and controls. In all tested groups, Mtb-hsp 16 concentration was higher than Mtb-hsp70 and Mtb-hsp 65. We suggest that lower levels of NOx may induce a M. tuberculosis genetic dormancy program via higher Mtb-hsp 16 expression in SA. It seems that Mtb-hsp 16 may be more important than Mtb-hsp70 and Mtb-hsp 65 in CIs formation and initiate an autoimmune response in SA related to mycobacteria's stationary-phase.

St. John's wort may relieve negative effects of stress on spatial working memory by changing synaptic plasticity.

Beneficial effects of St. John's wort (Hypericum perforatum) in the treatment of stress-evoked memory impairment were recently described. In this study, we tested a hypothesis that St. John's wort alleviates stress- and corticosterone-related memory impairments by restoring levels of synaptic plasticity proteins: neuromoduline (GAP-43) and synaptophysin (SYP) in hippocampus and prefrontal cortex. Stressed and corticosterone-treated rats displayed a decline in the acquisition of spatial working memory (p < 0.001) in the Barnes maze (BM). Chronic administration of H. perforatum (350 mg kg(-1) for 21 days), potently and significantly improved processing of spatial information in the stressed and corticosterone-injected rats (p < 0.001). Also, St Johns' wort statistically significantly (p < 0.05) increased levels of GAP-43 and SYP, respectively in the hippocampi and prefrontal cortex as measured by western immunoblotting. We found that H. perforatum prevented the deleterious effects of both chronic restraint stress and prolonged corticosterone administration on working memory measured in the BM test. The herb significantly (p < 0.01) improved hippocampus-dependent spatial working memory in comparison with control and alleviated some other negative effects of stress on cognitive functions. These findings increase our understanding of the reaction of the hippocampus and prefrontal cortex to stressful assaults and provide new insight into the possible actions of H. perforatum in the treatment of patients with impaired adaptation to environmental stressors and simultaneously suffering from cognitive impairment.

[Treatment of chronic obstructive pulmonary disease--traditional bronchodilatation and targeted anti-inflammatory therapy]. / Leczenie przewleklej obturacyjnej choroby pluc--bronchodylatacja i przeciwzapalna farmakoterapia celowana.

Currently available pharmacological treatment of COPD relies mostly on prophylaxis (smoking cessation) and symptomatic treatment, i.e. inhaled anticholinergic agents, ß2-agonists and phosphodiesterase inhibitors, aiming in their bronchodilatation capacity. Inhaled corticosteroid therapy is mainly prescribed in far advanced stages of the disease and its role in disease modification is still controversial. The authors analize currently available treatment modalities with regards to their potential anti-inflammatory and pleiotropic mode of action, which may lead to disease course modification.

Tiotropium increases cytosolic muscarinic M3 receptors and acetylated H3 histone proteins in induced sputum cells of COPD patients.

OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is characterized by irreversible progressive airflow limitation related to tobacco smoking. This limitation is caused by chronic inflammation of the airways and lung parenchyma and is associated with increased activity of parasympathetic system. The most effective bronchodilators in COPD are muscarinic receptor antagonists (MRA), which reverse, at least in part, compromised respiratory function. MRA also contribute to control inflammatory processes via interactions with inflammatory signaling molecules. The use of the long-acting cholinolytic bronchodilatator - tiotropium, with high affinity to M3 receptors, is suggested as a first line maintenance treatment in COPD patients. MATERIAL AND METHODS: In this study we assessed M3 receptor protein expression in induced sputum of 27 stable COPD patients before and after therapy consisting of 18 µg once daily tiotropium for 12 weeks. Lung function tests including spirometry, lung volumes, and DLCO were performed before and after therapy in all COPD patients. The patients were subjected to the sputum induction procedures before and after therapy. Sputum cells were isolated, sample-specific cell profiles were characterized, and the cells were processed to isolate pure cytosolic fractions. Cytosolic M3 protein and HDAC2 levels and nuclear acetylated histone H3 (AcH3) expression was quantified using specific antibodies against human proteins and Western blot with enhanced luminescence detection. RESULTS: Therapy significantly increased the mean forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) volume (P<0.05). The mean expression of M3 protein was higher by 37% after therapy (P<0.05), HDAC2 expression was not altered, while AcH3 level was increased by about 90% (P<0.01), compared with the corresponding data before therapy. HDAC2 expression before therapy was positively correlated with AcH3 expression (r = 0.74), while after therapy no correlation was detected. FEV1, FCV, and cytosolic M3 protein expression did not correlate with other biochemical parameters tested. CONCLUSIONS: Twelve weeks of tiotropium therapy in COPD patients improves clinical indices of lung function and involves alterations in sputum cell chromatin acetylation and also increased cholinergic M3 receptor internalization.

Activation of CREB by St. John's wort may diminish deletorious effects of aging on spatial memory.

St. John's wort (Hypericum perforatum) is one of the leading psychotherapeutic phytomedicines. Beneficial effects of this herb in the treatment of mild to moderate depression are well known. In this study we tested a hypothesis that St. John's wort alleviates age-related memory impairments by increasing the levels of cyclic adenosine 3', 5'-monophosphate response element binding protein (CREB) and phosphorylated CREB (pCREB) in hippocampus. Middleaged rats (18 month-old) displayed a decline in the acquisition of spatial working memory (p < 0.001) in the Morris water maze (MWM). Chronic administration of Hypericum perforatum (HP) (350 mg/kg for 21 days), potently and significantly improved the processing of spatial information in the aged rats (p < 0.001). Also the herb increased the levels of pCREB in the aged rat's hippocampus (p < 0.01) as measured by western immunoblotting. Aging caused significant locomotor impairments as tested in the open field (p < 0.001) but not in the MWM test. However, these were unaffected by treatment with HP. Thus, this study indicates that St. John's wort effectively prevents aging-induced deterioration of spatial memory in 18 month-old rats, possibly by the activation of CREB regulated genes associated with memory formation. It appears that mechanism is probably inactive in young rats.

Cerebrospinal fluid oxidative stress during chemotherapy of acute lymphoblastic leukemia in children.

In this study the authors addressed the question whether neurotoxicity due to the chemotherapy of acute lymphoblastic leukemia (ALL) is associated with cerebrospinal fluid (CSF) oxidative stress. Examination of 38 ALL patients revealed significant increases in 8-isoprostane concentration and important decreases in total antioxidative capacity of CSF during therapy. The mean 8-isoprostane level at diagnosis was 9.05 +/- 1.62 pg/mL, and no correlations with initial leukocytosis, organomegaly, and lactate dehydrogenase levels were noted. 8-Isoprostane concentrations were increased on the 59th day of treatment (mean level: 24.85 +/- 7.59 pg/mL [P < .01]) and remained elevated at 4 points of the consolidation phase (17.28 +/- 2.16 pg/mL [P < .05]; 22.72 +/- 6.04 pg/mL [P < .05]; 24.92 +/- 6.31 pg/mL [P < .01]; 32.32 +/- 7.94 pg/mL [P < .01]) as compared to their level at diagnosis. The mean total antioxidative capacity at diagnosis was 203.08 +/- 6.17 mumol/L and was remarkably decreased on the 59th day of treatment (189.76 +/- 1.9 mumol/L [P < .05]) and at one point of the consolidation phase (188.29 +/- 3.46 mumol/L [P < .05]) as compared to the level at diagnosis. This study indicates that neurotoxicity of standard ALL treatment may be related to oxidative stress.

Negative correlation between cerebrospinal fluid tau protein and cognitive functioning in children with acute lymphoblastic leukemia.

The aim of the study was to assess whether cerebrospinal fluid tau protein is associated with cognitive changes in children with acute lymphoblastic leukemia (ALL). Examination of 38 ALL patients revealed a statistically significant increase in tau protein on treatment day 59 and at two points during consolidation phase. Cognitive functioning was examined in 19 patients at an average of 3.7 years after diagnosis. The level of tau at the initiation of maintenance therapy was negatively correlated with verbal abilities measured on an intellectual scale. The study suggests that standard ALL treatment may cause a decline in cognitive functioning.

Ruthenium red protects HepG2 cells overexpressing CYP2E1 against acetaminophen cytotoxicity.

We studied the mechanisms of acetaminophen (APAP) cytotoxicity in HepG2 cells overexpressing cytochrome p4502E1, particularly the role of oxidative/nitrosative stress and ryanodine Ca2+ channel. Cells were grown for 24 h with APAP in the presence or absence of 4-methylpyrazole (4MP), L-arginine methyl ester (L-NAME), superoxide dismutase (SOD), or ruthenium red (RuR). Drug cytotoxicity was also tested in cells pretreated overnight with V-PYRRO/NO. APAP was without effect on empty vector-transfected cells, but damaged CYP2E1-transfected cells and this was abolished by RuR, reduced by 4MP, or V-PYRRO/NO but affected by L-NAME or SOD. APAP increased microsomal [3H]-ryanodine binding, while microsomal Ca2+ uptake was significantly lowered. RuR increased net microsomal Ca2+ uptake and normalized cytosolic Ca2+ levels. We can conclude that neither oxidative nor nitrosative stress is relevant to APAP cytotoxicity in cultured HepG2 cells, but our results point to ryanodine receptors as a potential crucial protein in the early stages of APAP cytotoxicity.