BSA-binding properties and anti-proliferative effects of amino acids functionalized polyoxomolybdates

Glycine decorated heteropolymolybdates, K₂Na[AsMo₆O₂₁(O₂CCH₂NH₃)₃]·6H₂O 1 and K₂Na₂[γ-Mo₈O₂₆(O₂CCH₂NH₃)₂]·6H₂O 2, have been synthesized and evaluated for in vitro anti-proliferative effects. The identity and high purity of compounds 1 and 2 were confirmed by elemental analysis, FT-IR spectrum, UV–vis spectrum, and X-ray diffraction. Crystal data for 2: triclinic, P-1, a = 9.792(2) Å, b = 10.077(2) Å, c = 10.351(2) Å, α = 83.865(4)°, β = 71.110(4)°, γ = 62.284(3)°, V = 854.3(3) Å³, Z = 1, R(final) = 0.0486. The inhibitory effects of 1 and 2 on human non-small cell lung cancer cell line A549 were investigated by MTT assay, nuclear staining, and the flow cytometry. It indicated that compound 1 inhibited the proliferation of A549 cells in a dose-dependent and time-dependent manner, which is more effective than the positive control, 5-fluorouracil (5-FU) (P < 0.05). The staining and flow cytometry results showed that compound 1 induced the apoptosis and necrosis of A549 cells and inhibit cell proliferation, which is associated with S-phase arrest. Compound 2 showed a modest activity in a dose-dependent manner. In addition, the interaction between compound 1 and bovine serum albumin (BSA) was evaluated by spectroscopic methods. The results showed that the compound 1 effectively quenched the intrinsic fluorescence of BSA via static quenching and changed the conformation of BSA.     

Enhanced cortical reperfusion protects coagulation factor XII-deficient mice from ischemic stroke as revealed by high-field MRI

Intrinsic coagulation factor XII deficient (FXII^?/?) mice are protected from ischemic stroke. To elucidate underlying mechanisms we investigated the early ischemic period in vivo by multimodal magnetic resonance imaging (MRI) at 17.6 Tesla.Cerebral ischemia was induced by either transient (60 min) or permanent occlusion of the middle cerebral artery (t/pMCAO). 10 FXII^?/? mice underwent t- , 10 FXII^?/? mice p- and 10 Wildtype (Wt) mice tMCAO. Cerebral blood flow (CBF), diffusion-weighted-imaging (DWI) and T2-relaxometry were measured at 2 h and 24 h after MCAO. Outcome measures were evaluated after motion correction and normalization to atlas space. 2 h after tMCAO CBF reduction was similar in FXII^?/? and Wt mice extending over cortical (CBF (ml/100 g/min) 33.6 ± 6.9 vs. 35.3 ± 4.6, p = 0.42) and subcortical regions (25.7 ± 4.5 vs. 31.6 ± 4.0, p = 0.17). At 24 h, recovery of cortical CBF by +36% was observed only in tMCAO FXII^?/? mice contrasting a further decrease of – 30% in Wt mice after tMCAO (p = 0.02, F_(1,18) = 6.24). In FXII^?/? mice in which patency of the MCA was not restored (pMCAO) a further decrease of ? 75% was observed. Cortical reperfusion in tMCAO FXII^?/? mice was related to a lower risk of infarction of 59% vs. 93% in Wt mice (p = 0.04). Subcortical CBF was similarly decreased in both tMCAO groups (Wt and FXII^?/?) relating to a similar risk of infarction of 89% (Wt) vs. 99% (FXII^?/?, p = 0.17).Deficiency of FXII allows neocortical reperfusion after tMCAO and rescues brain tissue by this mechanism. This study supports the concept of FXII as a promising new target for stroke prevention and therapy.     

Proatherogenic disturbances in lipoprotein profile,associated enzymes and transfer proteins in women with iron deficiency anaemia

ObjectiveTo characterize the lipid-related atherogenic risk factors in iron deficiency anaemia (IDA) patients.Design and methodsTwenty IDA women were compared to healthy age-matched controls. Lipoprotein profile, cholesteryl ester transfer protein (CETP), paraoxonase (PON) 1 and lipoprotein-associated phospholipase A₂ (LpPLA₂) activities and plasma levels of oxidized-LDL were evaluated.ResultsTriglycerides were higher (median [range]) (1.0 [0.5–1.9] vs. 0.7 [0.5–1.5] mmol/L, p < 0.05) and HDL-C lower (mean ± SD) (1.3 ± 0.3 vs. 1.6 ± 0.4 mmol/L, p < 0.01) in the patients group. CETP (197 ± 29% vs. 151 ± 29% mL^? 1 h^? 1, p < 0.001), PON 1 (122 ± 17 vs. 140 ± 33 μmol mL^? 1 min^? 1, p < 0.05) and LpPLA₂ (9.6 ± 2.0 vs. 8.1 ± 1.7 μmol mL^? 1 h^? 1, p < 0.05) activities were different in IDA women. No difference was observed in oxidized-LDL. Haemoglobin correlated negatively with triglycerides (r = ? 0.35, p < 0.05), CETP (r =  ?  0.62, p < 0.001) and LpPLA₂ (r =  ?  0.34, p < 0.05), while ferritin was positively associated with HDL-C (r =  0.39, p < 0.05) and inversely with CETP (r =  ?  0.49, p < 0.005).ConclusionThe alterations in lipoprotein profile, CETP, PON 1 and LpPLA₂ activities described in the present study indicate that non-treated IDA might represent a proatherogenic state.     

Synthesis and photodynamic activity of unsymmetrical A3B tetraarylporphyrins functionalized with l-glutamate and their Zn(II) and Cu(II) metal complex derivatives

Four novel unsymmetrical A₃B porphyrins 1, 2, 3 and 4 were synthesized following Lindsey procedure. Porphyrins 3 and 4 include one and three l-glutamate groups, respectively, and all porphyrins were metallated with Zn(II) (1a–4a) or Cu(II) (1b–4b). Porphyrins and metalloporphyrins presented values of singlet oxygen quantum yields (ΦD) ranging from 0.21 to 0.67. The tetraaryl derivatives in this study showed phototoxicity in SiHa cells with IC₅₀ values ranging from <0.01 to 6.56 ± 0.11 μM, the metalloporphyrin 4a showed the lowest IC₅₀ value. Comparing the phototoxic activity between all porphyrins, functionalization of porphyrins with glutamate increased 100 times phototoxic activity (1 (IC₅₀ 4.81 ± 0.34 μM) vs. 3 (IC₅₀ 0.04 ± 0.02 μM) and 2 (IC₅₀ 5.19 ± 0.42 μM) vs. 4 (IC₅₀ 0.05 ± 0.01 μM)). This increased activity could be attributed to reduced hydrophobicity and increased ΦΔ, given by functionalization with l-glutamate. Metalloporphyrins 3a (IC₅₀ 0.04 ± 0.01 μM) and 4a (IC₅₀ <0.01 μM) presented the best values ​​of phototoxic activity. Therefore, functionalization and zinc metalation increased the phototoxic activity. SiHa cells treated with porphyrins 3, 4, 3a and 4a at a final concentration of 10 μM, showed increased activity of caspase-3 enzyme compared to the negative control; indicating the induction of apoptosis. Differential gene expression pattern in SiHa cells was determined; treatments with metalloporphyrins 4a and 4b were performed, respectively, comparing the expression with untreated control. Treatments in both cases showed similar gene expression pattern in upregulated genes, since they share about 25 biological pathways and a large number of genes. According to the new photophysical properties related to the structural improvement and phototoxic activity, these molecules may have the potential application as photosensitizers in the photodynamic therapy.     

Detection of serum β2-GPI–Lp(a) complexes in patients with systemic lupus erythematosus

BackgroundCirculating β₂-glycoprotein-I-oxidized low-density lipoprotein (β₂-GPI–ox-LDL) complexes have been found in patients with systemic lupus erythematosus (SLE) and other autoimmune diseases as a contributor to the development of autoimmune-mediated atherosclerosis. In vitro study showed that β₂-GPI also bound with high affinity to atherogenic lipoprotein (a) [Lp(a)] which shares structural similarity to LDL. We examined the existence and clinical significance of serum complexes of β₂-GPI with Lp(a) in SLE patients.MethodsA “sandwich” ELISA was developed for measuring serum concentrations of β₂-GPI–Lp(a) complexes, using rabbit anti-human β₂-GPI antibody as capturing antibody, and quantitating with antibody against apo(a). Forty-seven SLE patients and 42 healthy controls were studied.ResultsBoth Lp(a) (400 ± 213 mg/l vs. 181 ± 70 mg/l) and ox-Lp(a) (27.07 ± 22.30 mg/l vs. 8.20 ± 4.55 mg/l) concentrations were higher in SLE patients than in controls (P < 0.0001). β₂-GPI–Lp(a) complexes were detectable in both controls and SLE. The complexes levels in SLE were higher than in controls (0.96 ± 0.41 U/ml vs. 0.59 ± 0.20 U/ml, P < 0.0001) and was positively correlated with ox-Lp(a) (P < 0.001).ConclusionsWe report the existence of β₂-GPI–Lp(a) complexes in both controls and SLE patients. The complexes levels increase in SLE.     

Paraoxonase activity in athletes with depleted iron stores and iron-deficient erythropoiesis

ObjectiveThe 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 methodsBased 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.ResultsSignificant 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, η² = 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 (χ² = 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.ConclusionsDeficient erythropoiesis in athletes contributes to impaired PON1 activity. In contrast, iron depletion, regardless of increased oxidative stress, does not affect PON1 activity.     

Increased low-density lipoprotein oxidation,but not total plasma protein oxidation,in Alzheimer's disease

ObjectivesThe two most common forms of dementia are Alzheimer's disease (AD), and vascular dementia (VaD). In the overlap of biochemical processes which have been identified in AD and VaD, oxidative stress is believed to contribute to the numerous pathologies of both dementias.Design and methodsThis study assessed oxidative damage in total plasma proteins, and isolated LDL in AD patients and age matched controls, in addition total antioxidant capacity (TAC) was measured.ResultsSignificantly higher LDL protein carbonylation was observed in AD compared to age-matched controls (AD: 4.17 ± 0.73 vs. control: 3.85 ± 0.86 nmol/mg LDL; p = 0.05, 2-tailed Mann–Whitney), in addition to reduced TAC (AD: 924.708 ± 174.429 vs. control: 1078.536 ± 252.633 μM; p = 0.001, 2-tailed Mann–Whitney). No differences were seen in total plasma protein carbonyl content (AD: 3.88 ± 0.31 vs. control: 3.98 ± 0.48 nmol/mg protein).ConclusionThe results further support the view that oxidation events in AD may be specific in nature, and represent functional changes to proteins, rather than random global events.     

Long-term stability of soluble ST2 in frozen plasma samples

ObjectiveThe aim of this study was to investigate the long-term in vitro stability of soluble ST2 (sST2).Design and methodsEDTA plasma samples were drawn from 15 individuals with various diseases. The Presage^TM ST2 assay was used for measurement of sST2 concentrations directly after blood collection and after storing plasma samples for 18 months at ?20 °C and ?80 °C. The default criterion for analyte stability was set at 95%.ResultssST2 concentrations in the 15 individuals ranged from 12 U/mL to 140 U/mL. Directly after blood collection, the mean (± SD) sST2 concentration was 51 ± 37 U/mL, and absolute analyte recoveries were 50 ± 35 U/mL and 51 ± 34 U/mL after storage of samples for 18 months at ?20 °C and ?80 °C, respectively. Relative analyte recoveries after 18 months of storage at ?20 °C and ?80 °C were 99 ± 5% and 101 ± 7%.ConclusionsST2 is stable for at least 1.5 years in plasma samples stored at ?20 °C and ?80 °C.     

Human plasma concentrations of malondialdehyde (MDA) and the F2-isoprostane 15(S)-8-iso-PGF2α may be markedly compromised by hemolysis: Evidence by GC-MS/MS and potential analytical and biological ramifications

Objectives:Malondialdehyde (MDA) and the F₂-isoprostane 15(S)-8-iso-prostaglandin F_2α (15(S)-8-iso-PGF_2α) belong to the most frequently analyzed biomarkers of oxidative stress in basic and clinical research. The objective of the present study was to examine the effect of hemolysis on free MDA and total (free + esterified) 15(S)-8-iso-PGF_2α concentrations in human plasma.Design and methods:MDA and 15(S)-8-iso-PGF_2α were determined by GC-MS/MS in plasma samples from venous heparinized blood drawn under resting conditions (n = 22) as well as under physical exercise (n = 158) in 22 healthy young subjects. In vitro, we prepared plasma samples with hemolysis degrees up to 0.8% using artificially hemolyzed, freshly obtained heparinized blood.Results:In some plasma samples of the exercise study both under resting and exercise conditions, clinically significant hemolysis was macroscopically visible. Both in vivo (r = 0.74) and in vitro (r = 0.87), we found a significant positive correlation between hemolysis degree (0–0.2%) and MDA plasma concentrations (50–250 nmol/L). Unlike in vitro (r = 0.84), in vivo, 15(S)-8-iso-PGF_2α and MDA plasma concentrations correlated weakly (r = 0.50).Conclusions:We hypothesize that free hemoglobin catalyzes the formation of MDA and 15(S)-8-iso-PGF_2α from free and esterified arachidonic acid. Plasma concentrations of MDA and total 15(S)-8-iso-PGF_2α may be markedly compromised by hemolysis. Measurements of MDA and 15(S)-8-iso-PGF_2α should be treated with caution regarding involvement of oxidative stress in disease as well as in health both under resting conditions and under exercise.     

Glutamate dehydrogenase activity in lymphocytes of B-cell chronic lymphocytic leukaemia patients

Objectives: To investigate the pattern of glutamate dehydrogenase (GLDH) activity, GLUD1 and GLUD2 expressions in peripheral blood mononuclear cells (PBMC) of untreated B-chronic lymphocytic leukemia (B-CLL) in healthy individuals (HI) and patients with infectious mononucleosis (IM).Design and methods: GLDH activity was determined in a supernatant obtained from pelleted PBMC. GLUD1 and GLUD2 mRNA expression was determined using a quantitative real-time polymerase chain reaction. CD19⁺ B cells from PBMC were purified by using positive selection.Results: The highest GLDH activity was found in PBMC of the B-CLL group followed by the HI group and IM group. The PBMC GLDH activity was higher in 60% of the B-CLL patients according to the established reference interval for our HI (2.17–5.70 μkat/g protein). The greater GLDH activity was also found in the CD19⁺ cell preparation of the B-CLL patients (two of the three) but not in HI (n = 3). The median value of GLUD1 expression was highest in the IM group (n = 11), followed by the HI (n = 14) and B-CLL groups (n = 59) (median 4.69/3.78, P < 0.005 and 4.69/2.91, P < 0.0005, respectively). GLUD2 expression was not significantly different between groups.Conclusions: The increased GLDH activity is specific for the PBMC of B-CLL patients. The GLUD1 but not the GLUD2 gene expression pattern is different between the PBMC of IM and B-CLL patients.     

Development and validation of a rapid liquid chromatography isotope dilution tandem mass spectrometry (LC-IDMS/MS) method for serum creatinine

Objectives:To develop an isotope dilution liquid chromatography tandem mass spectrometry (LC-IDMS/MS) method for the standardization of serum creatinine.Design and methods:Supernatants obtained by protein precipitation were injected into a LC-MS/MS. Chromatography was performed on a cation exchanger pre-column in normal phase isocratic mode. Creatinine and creatinine-D₃ were quantified using ion transitions of m/z 114→44 and 117→47, respectively.Results:The method was calibrated with the NIST Standard Reference Material 914a and was found to be exact in analyzing the certified reference material SRM 967 from NIST (97.1 ± 0.9% and 102.1 ± 0.9% of target value for levels 1 and 2, respectively) and by calculating recuperation of spiked creatinine in 10 different patient samples (103.6 ± 4.1%). Intra-assay imprecision was 0.9% at both 64.6 and 354 μmol/L creatinine, while inter-assay imprecisions were 1.9% and 1.8%. Absence of ion suppression was confirmed by spiking experiments. The method was shown to be free of carryover. A good correlation was obtained between the LC-MS/MS method and a Jaffe method run on an automated analyzer (r = 0.999).Conclusions:We have developed a fast and simple method for the quantification of serum creatinine by isotope dilution tandem mass spectrometry (IDMS/MS) and we propose that this method can be used as a reference method by laboratories that wish to validate their serum creatinine automated assay.     

Antioxidant status and lipid peroxidation in the blood of breast cancer patients of different ages after chemotherapy with 5-fluorouracil,doxorubicin and cyclophosphamide

ObjectivesBreast carcinoma is related to the increase of lipid peroxidation in plasma with concomitant decrease of antioxidant (AO) defense capacity in blood cells, which becomes more pronounced during aging of the patients. This work evaluated the potential age-related effect of chemotherapy with 5-fluorouracil, doxorubicin and cyclophosphamide (FAC) on the level of lipid hydroperoxides (LP), glutathione (GSH), AO enzyme activities of copper, zinc superoxide dismutase (CuZnSOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) in breast cancer patients. The level of CuZnSOD protein was assessed after the FAC therapy and radiotherapy of breast cancer.Design and methodsAO parameters were measured in the blood of 58 breast cancer patients and 60 healthy age-matched healthy subjects by biochemical and Western blot analyses.ResultsIncreased oxidative stress (LP: p < 0.05) and decreased AO enzyme activities (CuZnSOD: p < 0.01, GPx: p < 0.05, GR: p < 0.01) and GSH level (p < 0.01) in the blood of breast cancer patients in response to FAC chemotherapy seem not to be age-dependent. CuZnSOD enzyme expression decreased after the FAC chemotherapy (p < 0.05), while it increased after the radiotherapy of breast cancer (p < 0.05).ConclusionFAC chemotherapy and radiotherapy promote further oxidative shift, which potentiate already existing chronic oxidative stress linked to breast cancer. In these effects, impaired capacity for H₂O₂ detoxification (CAT, GPX and GSH) seems to have major contribution.     

The association between transforming growth factor β3 polymorphisms and left ventricular structure in hypertensive subjects

BackgroundTransforming growth factor β (TGF-β) may be a crucial regulator of cardiac remodeling. We investigated the association between the TGF-β gene polymorphisms and left ventricular structure.MethodsA total of 658 hypertensive subjects were genotyped for the TGF-β1 T869C and TGF-β3 (rs3917187 and rs4252338) polymorphisms.ResultsTGF-β3 rs3917187 AA homozygotes had, while accounting for covariates, greater left ventricular end-systolic (LVESD, P = 0.004) and end-diastolic dimension (LVEDD, P = 0.007) than G allele carriers. Moreover, left ventricular mass index (LVMI) in AA genotype was 123.0 ± 3.1 g/m² significantly higher than that in AG (114.6 ± 1.6 g/m²) and GG (115.4 ± 2.1 g/m², P = 0.03) genotypes. In multivariate regression analysis, TGF-β3 rs3917187 genotype as an independent predictor had statistically significant effects on LVESD (β = 0.164, P = 0.002), LVEDD (β = 0.172, P = 0.003) and LVMI (β = 0.136, P = 0.016), respectively. In further analyses, we observed a significant interaction between the rs3917187 and alcohol intake in relation to LVESD (P_int = 0.04) and left ventricular fractional shortening (LVFSH, P_int = 0.012). However, no relationship could be found between left ventricular parameters and the T869C or the rs4252338.ConclusionThe present results demonstrated that the TGF-β3 rs3917187 polymorphism was associated with left ventricular structure, and had an interactive influence with alcohol on LVESD and LVFSH in hypertensive subjects.     

Stability of plasma adrenocorticotrophic hormone (ACTH): Influence of hemolysis,rapid chilling,time, and the addition of a maleimide

ObjectivesThe aim of this study was to examine the effects of hemolysis, rapid chilling, time, and the addition of a maleimide on the stability of human plasma ACTH measurements.Design and methodsPartially hemolyzed EDTA blood (n = 10), initially at 37 °C, was centrifuged at 4 °C either immediately or after rapid chilling in ice/water. Plasma ACTH was then measured either immediately, or after 1 h at 22 °C with or without the addition of 2 mM N-phenyl maleimide (NPM).ResultsFor 0.2% hemolysis compared to no hemolysis, the mean (±SEM) loss with immediate centrifugation and immediate ACTH measurement was 11 ± 1%. This loss was significantly (p < 0.002) reduced to 6 ± 1% by an initial rapid chilling of the samples. For analysis after 1 h at 22 °C, the addition of NPM decreased the loss of ACTH from 15 ± 2% to 2 ± 2% (p < 0.002).ConclusionRapid chilling, prompt analysis, and addition of NPM can each reduce the interference of hemolysis in the measurement of plasma ACTH concentrations.     

MP2RAGE,a self bias-field corrected sequence for improved segmentation and T1-mapping at high field

The large spatial inhomogeneity in transmit B₁ field (B₁⁺) observable in human MR images at high static magnetic fields (B₀) severely impairs image quality. To overcome this effect in brain T₁-weighted images, the MPRAGE sequence was modified to generate two different images at different inversion times, MP2RAGE. By combining the two images in a novel fashion, it was possible to create T₁-weigthed images where the result image was free of proton density contrast, T₂? contrast, reception bias field, and, to first order, transmit field inhomogeneity.MP2RAGE sequence parameters were optimized using Bloch equations to maximize contrast-to-noise ratio per unit of time between brain tissues and minimize the effect of B₁⁺ variations through space. Images of high anatomical quality and excellent brain tissue differentiation suitable for applications such as segmentation and voxel-based morphometry were obtained at 3 and 7 T.From such T₁-weighted images, acquired within 12 min, high-resolution 3D T₁ maps were routinely calculated at 7 T with sub-millimeter voxel resolution (0.65–0.85 mm isotropic). T₁ maps were validated in phantom experiments. In humans, the T₁ values obtained at 7 T were 1.15 ± 0.06 s for white matter (WM) and 1.92 ± 0.16 s for grey matter (GM), in good agreement with literature values obtained at lower spatial resolution. At 3 T, where whole-brain acquisitions with 1 mm isotropic voxels were acquired in 8 min, the T₁ values obtained (0.81 ± 0.03 s for WM and 1.35 ± 0.05 for GM) were once again found to be in very good agreement with values in the literature.     

In vitro and in vivo antileishmanial properties of a 2-n-propylquinoline hydroxypropyl β-cyclodextrin formulation and pharmacokinetics via intravenous route

2-n-propylquinoline (2-n-PQ) had shown interesting in vivo antileishmanial activities after administration by oral route on leishmaniasis animal models. However, the lipophilic properties of this compound avoid its use by intravenous route, this route being indicated in cases of severe visceral leishmaniasis with vomiting. Thus, a 2-n-propylquinoline hydroxypropyl beta-cyclodextrin (2-n-PQ-HPC) formulation was set up in this aim. The formulation was active in vitro both on Leishmania donovani axenic and intramacrophage amastigotes with IC₅₀ values at 6.22 ± 0.82 μM and 20.01 ± 0.52 μM, respectively, without any toxicity on macrophages. 2-n-PQ-HPC exhibited similar activity on WT and drug-resistant parasites. Its in vitro interactions with antimonials, amphotericin B and miltefosine were found as additive both in axenic amastigotes and intramacrophage amastigotes. 2-n-PQ-HPC was not able to generate drug resistance after in vitro drug pressure since the resistance index was less than 4. 2-n-PQ-HPC was also active on the L. donovani/Balb/c mice model with an intravenous treatment regimen at 10 mg kg⁻¹ day⁻¹ on 10 consecutive days without hepatic, renal and blood toxicity. The pharmacokinetics of 2-n-PQ in rats showed that after an intravenous treatment of the formulation at 10 mg kg⁻¹, the plasma drug concentrations rapidly declined bi-exponentially with a half-life of 58.7 min and a total clearance of 18.63 l h⁻¹ kg⁻¹. The apparent volume of distribution was higher than the blood volume in rats, indicating that 2-n-PQ was well distributed in tissues, allowing parasite elimination. Such a formulation is worth of further antiparasitic and toxicological evaluations.     

MUC1 expression and its association with other aetiological factors and localization to mitochondria in preneoplastic and neoplastic gastric tissues

BackgroundThe molecular events that underlie the conversion of normal human gastric epithelium into adenocarcinoma are poorly understood. MUC1 overexpression and localization in mitochondria might confer cancer cells with attenuation of stress induced apoptosis. We studied MUC1 expression pattern, interaction with HSP70 and localization in mitochondria in preneoplastic and neoplastic human gastric tissues.MethodsImmunohistochemistry and Western blot were used to study MUC1 expression pattern and localization in mitochondria. Coimmunoprecipitation was used to study MUC1 interaction with HSP70. MUC1 expression was correlated with other causative features including erbB2 expression.ResultsMUC1 was expressed in 75.8% (147/194). MUC1 overexpression was detected in 50.0% (19/38 cases) dysplasia and 58.2% (32/55 cases) adenocarcinoma tissues. MUC1-CT–HSP70 interaction was seen in 71.66% (43/60 cases) and MUC1 localized to mitochondria in 33.33% (5/15) dysplasia samples and in 47.05% (8/17) adenocarcinoma samples. MUC1 expression showed significant association with smoking (χ² = 5.945; p < 0.015), alcohol consumption (χ² = 4.055; p < 0.044) and erbB2 positivity (χ² = 10.75; p < 0.001). MUC1 expression did not show appreciable association with age (χ² = 0.15; p < 0.698), sex (χ² = 0.22; p < 0.640) or Helicobacter pylori infection (χ² = 3.06; p < 0.080).ConclusionsSignificant correlation was found between MUC1 expression and smoking, alcohol and erbB2 expression. MUC1 showed aberrant expression in dysplasia and adenocarcinoma stages. MUC1 cytosolic tail was bound by HSP70 in all the stages but MUC1-CT was found to localize in mitochondria only in dysplasia and adenocarcinoma. MUC1-CT localization to mitochondria in dysplasia and adenocarcinoma might aid in the attenuation of epithelial stress response induced loss of polarity.     

A ventilation technique for oxygenation and carbon dioxide elimination in CPR: Continuous insufflation of oxygen at three levels of pressure in a pig model

AimPulmonary ventilation remains an important part of cardiopulmonary resuscitation, affecting gas exchange and haemodynamics. We designed and studied an improved method of ventilation for CPR, constructed specifically to support both gas exchange and haemodynamics. This method uses continuous insufflation of oxygen at three levels of pressure, resulting in tri-level pressure ventilation (TLPV). We hypothesized that TLPV improves gas exchange and haemodynamics compared to manual gold standard ventilation (GSV).MethodsIn 14 pigs, ventricular fibrillation was induced and automated CPR performed for 10 min with either TLPV or GSV. After defibrillation, CPR was repeated with the other ventilation method. Gas exchange and haemodynamics were monitored. Data are presented as mean ± standard error of the mean.ResultsTLPV was superior to GSV for PaO₂ (163 ± 36 mmHg difference; P = 0.001), and peak AWP (−20 ± 2 cmH₂O difference; P = 0.000) and higher for mean AWP (8 ± 0.2 cmH₂O difference; P = 0.000). TLPV was comparable to GSV for CPP (5 ± 3 mmHg difference; P = 0.012), VCO₂ (0.07 ± 0.3 mL/min/kg difference; P = 0.001), SvO₂ (4 ± 3%-point; P = 0.001), mean carotid flow (−0.5 ± 4 mL/min difference; P = 0.016), and pH_a (0.00 ± 0.03 difference; P = 0.002). The PaCO₂ data do not provide a conclusive result (4 ± 4 mmHg difference).ConclusionWe conclude that the ventilation strategy with a tri-level pressure cycle performs comparable to an expert, manual ventilator in an automated-CPR swine model.     

The mouth-to-bag resuscitator during standard anaesthesia induction in apnoeic patients

AimVentilation of a non-intubated emergency patient by inexperienced rescuers with a standard bag-valve device may result in high inspiratory flow rates and subsequently high airway pressures with stomach inflation. Therefore, a self-inflating bag has been developed that requires lay rescuers to blow up a single-use balloon inside an adult bag-valve device, which, in turn, displaces air within the bag towards the patient. This concept has been compared to standard adult bag-valve devices earlier in bench models but not in patients.MethodsAn anaesthetist who was blinded to all monitor tracings ventilated the lungs of 40 apnoeic patients during routine anaesthesia induction either with a standard bag-valve device or with the mouth-to-bag resuscitator in a random order. Study endpoints were peak inspiratory flow rates, peak airway pressure, tidal volumes and inspiratory time.ResultsPeak inspiratory flow was 40 ± 10 l min^?1 for the standard bag-valve device versus 33 ± 13 l min^?1 for the mouth-to-bag resuscitator (P < 0.0001); peak airway pressure was 17 ± 5 cmH₂O versus 14 ± 5 cmH₂O (P < 0.0001); inspiratory tidal volume was 477 ± 133 ml versus 644 ± 248 ml (P < 0.001) and inspiratory time was 1.1 ± 0.3 s versus 1.9 ± 0.6 s (P < 0.0001).ConclusionEmploying the mouth-to-bag resuscitator during simulated ventilation of a non-intubated patient in respiratory arrest significantly decreased peak inspiratory flow and peak airway pressure and increased inspiratory tidal volume and inspiratory times compared to a standard bag-valve device.     

Discovery of 2-(3,4-dichlorophenoxy)-N-(2-morpholin-4-ylethyl)acetamide: A selective σ1 receptor ligand with antinociceptive effect

Compound 2-(3,4-dichlorophenoxy)-N-(2-morpholin-4-ylethyl)acetamide (1) was designed, prepared and the in vitro binding evaluation against σ₁ and σ₂ receptors was measured. Compound 1 showed high σ₁ receptor affinity (K_i = 42 nM) and it was 36-times more selective for σ₁ than σ₂ receptor. Also, it was performed a molecular docking of compound 1 into the ligand binding pocket homology model of σ₁ receptor, showing a salt bridge between the ionized morpholine ring and Asp126, as well as important short contacts with residues Tyr120, His154 and Trp164. Ligand efficiency indexes and predicted toxicity analysis revealed an excellent intrinsic quality of 1. The antinociceptive effect of compound 1 was determined using the formalin test. The ipsilateral local peripheral (10–300 μg/paw) and intrathecal (100 μg/rat) administration of 1 produced a reduction in formalin-induced nociception. The in vivo results indicated that 1 may be effective in treating inflammatory pain.