Differential Expression of Circular RNAs in Glioblastoma Multiforme and Its Correlation with Prognosis

OBJECTIVE: The present study aimed to explore the expression profiles of circular RNAs (circRNAs) in glioblastoma multiforme (GBM) in an attempt to identify potential core genes in the pathogenesis of this tumor. METHODS: Differentially expressed circRNAs were screened between tumor tissues from five GBM patients and five normal brain samples using Illumina Hiseq. Bioinformatics analysis was used to analyze their potential function. CircBRAF was further detected in different WHO grades glioma tissues and normal brain tissues. Kaplan-Meier curves and multivariate Cox's analysis were used to analyze the association between circBRAF expression level and prognosis of glioma patients. RESULTS: A total of 1411 differentially expressed circRNAs were identified in GBM patients including 206 upregulated circRNAs and 1205 downregulated circRNAs. Differential expression of circRNAs was closely associated with the biological process and molecular function. The downregulated circRNAs were mainly associated with ErbB and Neurotrophin signaling pathways. Moreover, the expression level of circBRAF in normal brain tissues was significantly higher than that in glioma tissues (P < .001). CircBRAF was significantly lower in glioma patients with high pathological grade (WHO III & IV) than those with low grade (WHO I & II) (P < .001). Cox analysis revealed that high circBRAF expression was an independent biomarker for predicting good progression-free survival and overall survival in glioma patients (HR = 0.413, 95% CI 0.201-0.849; HR = 0.299, 95% CI 0.135-0.661; respectively). CONCLUSION: The present study identified a profile of dysregulated circRNAs in GBM. Bioinformatics analysis showed that dysregulated circRNAs might be associated with tumorigenesis and development of GBM. In addition, circBRAF could severe as a biomarker for predicting pathological grade and prognosis in glioma patients.     

Influence of Noise Correction on Intra- and Inter-Subject Variability of Quantitative Metrics in Diffusion Kurtosis Imaging

Diffusion kurtosis imaging (DKI) is a promising extension of diffusion tensor imaging, giving new insights into the white matter microstructure and providing new biomarkers. Given the rapidly increasing number of studies, DKI has a potential to establish itself as a valuable tool in brain diagnostics. However, to become a routine procedure, DKI still needs to be improved in terms of robustness, reliability, and reproducibility. As it requires acquisitions at higher diffusion weightings, results are more affected by noise than in diffusion tensor imaging. The lack of standard procedures for post-processing, especially for noise correction, might become a significant obstacle for the use of DKI in clinical routine limiting its application. We considered two noise correction schemes accounting for the noise properties of multichannel phased-array coils, in order to improve the data quality at signal-to-noise ratio (SNR) typical for DKI. The SNR dependence of estimated DKI metrics such as mean kurtosis (MK), mean diffusivity (MD) and fractional anisotropy (FA) is investigated for these noise correction approaches in Monte Carlo simulations and in in vivo human studies. The intra-subject reproducibility is investigated in a single subject study by varying the SNR level and SNR spatial distribution. Then the impact of the noise correction on inter-subject variability is evaluated in a homogeneous sample of 25 healthy volunteers. Results show a strong impact of noise correction on the MK estimate, while the estimation of FA and MD was affected to a lesser extent. Both intra- and inter-subject SNR-related variability of the MK estimate is considerably reduced after correction for the noise bias, providing more accurate and reproducible measures. In this work, we have proposed a straightforward method that improves accuracy of DKI metrics. This should contribute to standardization of DKI applications in clinical studies making valuable inferences in group analysis and longitudinal studies.     

Decreased expression of microRNA-200b is an independent unfavorable prognostic factor for glioma patients

Background and aimAs a member of the microRNA (miR)-200 family, miR-200b has been recognized as one of the fundamental regulators of epithelial–mesenchymal transition, chemosensitivity, cell proliferation, and cell cycle. Especially in glioma, miR-200b targets the CREB1 gene and suppresses the tumor cell growth in vitro. However, its involvement in human glioma has not yet been determined. The aim of this study was to investigate the clinical significance of miR-200b expression in this disease.MethodsmiR-200b expression in 266 pairs of human gliomas and matched nonneoplastic brain tissues was measured by real-time quantitative RT-PCR assay.ResultsCompared with nonneoplastic brain tissues, the expression level of miR-200b was significantly decreased in glioma tissues (tumor vs. normal: 2.87 ± 2.05 vs. 8.78 ± 2.50, P < 0.001). Of 266 patients with gliomas, 166 (62.41%) were in low miR-200b expression group. In addition, we found that the glioma tissues from high-grade tumors (grade III and IV) had much lower miR-200b expression than glioma tissues from low grade tumors (grade I and II). Moreover, the expression level of miR-200b was positively correlated with Karnofsky performance status (KPS) scores of glioma tissues. The results of a 60-month follow-up in 266 glioma patients further demonstrated that lower miR-200b expression was correlated with worse progression-free survival and overall survival in the patients with grade III and IV gliomas. Both univariate and multivariate analyses revealed that miR-200b was an independent prognostic indicator for glioma.ConclusionThese findings prove that the decreased expression of miR-200b may be associated with malignant tumor progression and poor prognosis in patients with gliomas, suggesting the potential role of miR-200b in glioma management. miR-200b may be a novel and valuable signature for predicting the clinical outcome of patients with gliomas.     

Specific PCR-based marker for detection of pathogenic groups of Fusarium oxysporum f. sp. cucumerinum in India

For the detection of Fusarium oxysporum f. sp. cucumerinum pathogenic groups, a specific PCR-based marker was developed. Specific random amplified polymorphic DNA (RAPD) markers which identified in four pathogenic groups I, II, III, and IV were cloned into P^Gem-Teasy vector. Cloned fragments were sequenced, and used for developing sequence characterized amplified regions (SCAR) primers for detection of pathogenic groups. F. oxysporum f. sp. cucumerinum isolates belonging to four pathogenic groups in India, cucumber nonpathogenic F. oxysporum, F. oxysporum f. sp. moniliforme and melonis, Fusarium udum, and isolate of Alternaria sp. were tested using developed specific primers. A single 1.320 kb, 770 bp, 1.119 kb, and 771 bp fragment were amplified from pathogenic group I, II, III, and IV isolates, respectively. Results showed the PCR based marker, which used in this research work, could detect up to 1 ng of fungal genomic DNA. The specific SCAR primers and PCR technique developed in this research easily detect and differentiate isolates of each F. oxysporum f. sp. cucumerinum pathogenic groups.     

Comparison between muscle activation measured by electromyography and muscle thickness measured using ultrasonography for effective muscle assessment

In this study, we aimed to compare the intrarater reliability and validity of muscle thickness measured using ultrasonography (US) and muscle activity via electromyography (EMG) during manual muscle testing (MMT) of the external oblique (EO) and lumbar multifidus (MF) muscles. The study subjects were 30 healthy individuals who underwent MMT at different grades. EMG was used to measure the muscle activity in terms of ratio to maximum voluntary contraction (MVC) and root mean square (RMS) metrics. US was used to measure the raw muscle thickness, the ratio of muscle thickness at MVC, and the ratio of muscle thickness at rest. One examiner performed measurements on each subject in 3 trials. The intrarater reliabilities of the % MVC RMS and raw RMS metrics for EMG and the % MVC thickness metrics for US were excellent (ICC = 0.81–0.98). There was a significant difference between all the grades measured using the % MVC thickness metric (p < 0.01). Further, this % MVC thickness metric of US showed a significantly higher correlation with the EMG measurement methods than with the others (r = 0.51–0.61). Our findings suggest that the % MVC thickness determined by US was the most sensitive of all methods for assessing the MMT grade.     

Role of damage-specific DNA polymerases in M13 phage mutagenesis induced by a major lipid peroxidation product trans-4-hydroxy-2-nonenal

One of the major lipid peroxidation products trans-4-hydroxy-2-nonenal (HNE), forms cyclic propano- or ethenoadducts bearing six- or seven-carbon atom side chains to G > C ? A > T. To specify the role of SOS DNA polymerases in HNE-induced mutations, we tested survival and mutation spectra in the lacZα gene of M13mp18 phage, whose DNA was treated in vitro with HNE, and which was grown in uvrA^? Escherichia coli strains, carrying one, two or all three SOS DNA polymerases. When Pol IV was the only DNA SOS polymerase in the bacterial host, survival of HNE-treated M13 DNA was similar to, but mutation frequency was lower than in the strain containing all SOS DNA polymerases. When only Pol II or Pol V were present in host bacteria, phage survival decreased dramatically. Simultaneously, mutation frequency was substantially increased, but exclusively in the strain carrying only Pol V, suggesting that induction of mutations by HNE is mainly dependent on Pol V. To determine the role of Pol II and Pol IV in HNE induced mutagenesis, Pol II or Pol IV were expressed together with Pol V. This resulted in decrease of mutation frequency, suggesting that both enzymes can compete with Pol V, and bypass HNE-DNA adducts in an error-free manner. However, HNE-DNA adducts were easily bypassed by Pol IV and only infrequently by Pol II.Mutation spectrum established for strains expressing only Pol V, showed that in uvrA^? bacteria the frequency of base substitutions and recombination increased in relation to NER proficient strains, particularly mutations at adenine sites. Among base substitutions A:T → C:G, A:T → G:C, G:C → A:T and G:C → T:A prevailed.The results suggest that Pol V can infrequently bypass HNE-DNA adducts inducing mutations at G, C and A sites, while bypass by Pol IV and Pol II is error-free, but for Pol II infrequent.     

Structured near-infrared Magnetic Circular Dichroism spectra of the Mn4CaO5 cluster of PSII in T. vulcanus are dominated by Mn(IV) d-d ‘spin-flip’ transitions

Photosystem II passes through four metastable S-states in catalysing light-driven water oxidation. Variable temperature variable field (VTVH) Magnetic Circular Dichroism (MCD) spectra in PSII of Thermosynochococcus (T.) vulcanus for each S-state are reported. These spectra, along with assignments, provide a new window into the electronic and magnetic structure of Mn₄CaO₅. VTVH MCD spectra taken in the S₂ state provide a clear g = 2, S = 1/2 paramagnetic characteristic, which is entirely consistent with that known by EPR. The three features, seen as positive (+) at 749 nm, negative (?) at 773 nm and (+) at 808 nm are assigned as ⁴A → ²E spin-flips within the d³ configuration of the Mn(IV) centres present. This assignment is supported by comparison(s) to spin-flips seen in a range of Mn(IV) materials. S₃ exhibits a more intense (?) MCD peak at 764 nm and has a stronger MCD saturation characteristic. This S₃ MCD saturation behaviour can be accurately modelled using parameters taken directly from analyses of EPR spectra. We see no evidence for Mn(III) d-d absorption in the near-IR of any S-state. We suggest that Mn(IV)-based absorption may be responsible for the well-known near-IR induced changes induced in S₂ EPR spectra of T. vulcanus and not Mn(III)-based, as has been commonly assumed. Through an analysis of the nephelauxetic effect, the excitation energy of S-state dependent spin-flips seen may help identify coordination characteristics and changes at each Mn(IV). A prospectus as to what more detailed S-state dependent MCD studies promise to achieve is outlined.     

Earthworm-mediated synthesis of silver nanoparticles: A potent tool against hepatocellular carcinoma,Plasmodium falciparum parasites and malaria mosquitoes

The development of parasites and pathogens resistant to synthetic drugs highlighted the needing of novel, eco-friendly and effective control approaches. Recently, metal nanoparticles have been proposed as highly effective tools towards cancer cells and Plasmodium parasites. In this study, we synthesized silver nanoparticles (EW–AgNP) using Eudrilus eugeniae earthworms as reducing and stabilizing agents. EW–AgNP showed plasmon resonance reduction in UV–vis spectrophotometry, the functional groups involved in the reduction were studied by FTIR spectroscopy, while particle size and shape was analyzed by FESEM. The effect of EW–AgNP on in vitro HepG2 cell proliferation was measured using MTT assays. Apoptosis assessed by flow cytometry showed diminished endurance of HepG2 cells and cytotoxicity in a dose-dependent manner. EW–AgNP were toxic to Anopheles stephensi larvae and pupae, LC₅₀ were 4.8 ppm (I), 5.8 ppm (II), 6.9 ppm (III), 8.5 ppm (IV), and 15.5 ppm (pupae). The antiplasmodial activity of EW–AgNP was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of Plasmodium falciparum. EW–AgNP IC₅₀ were 49.3 μg/ml (CQ-s) and 55.5 μg/ml (CQ-r), while chloroquine IC₅₀ were 81.5 μg/ml (CQ-s) and 86.5 μg/ml (CQ-r). EW–AgNP showed a valuable antibiotic potential against important pathogenic bacteria and fungi. Concerning non-target effects of EW–AgNP against mosquito natural enemies, the predation efficiency of the mosquitofish Gambusia affinis towards the II and II instar larvae of A. stephensi was 68.50% (II) and 47.00% (III), respectively. In EW–AgNP-contaminated environments, predation was boosted to 89.25% (II) and 70.75% (III), respectively. Overall, this research highlighted the EW–AgNP potential against hepatocellular carcinoma, Plasmodium parasites and mosquito vectors, with little detrimental effects on mosquito natural enemies.     

Changes in knee joint muscle activation patterns during walking associated with increased structural severity in knee osteoarthritis

PurposeTo determine whether alterations in knee joint muscle activation patterns during gait were related to structural severity determined by Kellgren–Lawrence (KL) radiographic grades, for those with a moderate knee OA classification.ScopeEighty-two individuals with knee OA, classified as moderate using a functional and clinical criterion were stratified on KL-grade (KL II, KL III and KL IV). Thirty-five asymptomatic individuals were matched for age and walking velocity. Lower limb motion and surface electromyograms from rectus femoris plus lateral and medial sites for the gastrocnemii, vastii and hamstring muscles were recorded during self-selected walking. Gait velocity and characteristics from sagittal plane knee angular displacement waveforms were calculated. Principal component analysis extracted amplitude and temporal features from electromyographic waveform. Analysis of variance models tested for main effects (group, muscle) and interactions (α = 0.05) for these features. No differences in anthropometrics, velocity, knee muscle strength and symptoms were found among the three OA groups (p > 0.05). Specific features from medial gastrocnemius, lateral hamstring and quadriceps amplitude and temporal patterns were significantly different among OA groups (p < 0.05).ConclusionsSystematic alterations in specific knee joint muscle activation patterns were associated with increasing structural severity based on KL-grades whereas other alterations were associated with the presence of OA.     

Angiotensin III stimulates high stretch-induced ANP secretion via angiotensin type 2 receptor

Angiotensin III (Ang III) is metabolized from Ang II by aminopeptidase (AP) A and in turn, Ang III is metabolized to Ang IV by APN. Ang III is known to have a similar effect to Ang II on aldosterone secretion, but the effect of Ang III on atrial natriuretic peptide (ANP) secretion from cardiac atria is not known. The aim of the present study is to define the effect of Ang III on ANP secretion and its receptor subtype using isolated perfused beating atria. The volume load was achieved by elevating the height of outflow catheter connected with isolated atria from 5 cmH₂O to 7.5 cmH₂O. Atrial stretch by volume load increased atrial contractility and ANP secretion. Ang III stimulated stretch-induced ANP secretion in a dose-dependent manner without change in atrial contractility. The stimulated effect of Ang III (1 μM) on stretch-induced ANP secretion was blocked by the pretreatment of Ang II type 2 (AT₂) receptor antagonist but not by AT₁ or Mas receptor antagonist. Pretreatment with inhibitor of phosphoinositide 3-kinase (PI3K), Akt, nitric oxide synthase, soluble guanylyl cyclase, or protein kinase G (PKG) attenuated Ang III-stimulated ANP secretion. When Ang III (40 nM) or Ang II (4 nM) was infused for 10 min into anesthetized rats, mean arterial pressure was increased about 10%. However, Ang III increased plasma ANP level by 35.81 ± 10.19% but Ang II decreased plasma ANP level by 30.41 ± 7.27%. Therefore, we suggest that Ang III, opposite to Ang II, stimulated stretch-induced ANP secretion through AT₂ receptor/PI3K/Akt/nitric oxide/PKG pathway.     

Chemical composition,antioxidant and macromolecule damage protective effects of Picrorhiza kurroa Royle ex Benth

In the present study, we identified the chemical constituents of 70% hydroalcoholic fraction of Picrorhiza kurroa by LC–ESI–MS/MS which showed the presence of iridoid glucosides such as picroside I, picroside II, picroside III, picroside IV, kutkoside, pikuroside and flavonoids like apocynin and vanillic acid. P. kurroa exhibited DPPH radical scavenging and metal chelating activities with IC₅₀ of 75.16 ± 3.2 and 55.5 ± 4.8 μg/mL and also showed potent reducing power and total antioxidant activities. The extract inhibited macromolecule damage such as H₂O₂ induced plasmid DNA damage and AAPH induced oxidation of bovine serum albumin and lipid peroxidation of rat hepatic tissues.     

The protective effect of montelukast sodium on carbon tetrachloride induced hepatopathy in rat

AimThis study investigates the effects of montelukast sodium (MK) (CysLTLT1 receptor antagonist) on CCl₄induced hepatopathy on rat.Material and methodsWe worked on 4 groups of 10 Wistar male rats each. The groups received as follows: group I (control group) – saline, group II – MK 5 mg/kg/day i.p. for 5 days, group III – MK 5 mg/kg/day i.p., 1 day prior to and 4 days concomitantly with CCl₄ p.o., 0.3 ml/Kg/day and group IV – CCl₄, p.o., 0.3 ml/Kg/day for 4 days. One day after the last administration, samples of blood were taken and alanine aminotransferase (ALT), total bilirubin (TB), direct bilirubin (DB), malondialdehyde (MDA), catalase (CAT) as well as total antioxidant capacity (TAC) were determined. The histopathological exam was performed. We also determined superoxide dismutase (SOD), MDA, CAT and GSH in liver homogenate.ResultsCompared to group IV, group III exhibited statistically significant lower levels of ALT (318 ± 15.75 versus 203.14 ± 10.28 UI, p < 0.0001), TB (3.16 ± 0.30 versus 1.99 ± 0.08 mg/dl, p < 0.0001), MDA in blood and in liver homogenate (4.98 ± 1.71 versus 2.15 ± 1.18 nmol/ml, p = 0.0004) and higher levels of SOD and CAT. Histopathologically, group IV presented important macro- and micro-vesicular hepatic steatosis and group III preserved lobular histoarchitecture and had less severe cellular lesions.ConclusionMK exhibits a partial hepatoprotective effect on rats treated with CCl₄.     

Embryo production and progesterone profiles in ewes superovulated with different hormonal treatments

The superovulatory response and embryo yield following hormonal treatments of Merino ewes during late spring and their estrous cycle were evaluated. Ewes (n=17) were treated with progestagen-impregnated sponges and assigned to Group I (800 IU PMSG plus 11.5 mg FSH-p); Group II (1200 IU PMSG); Group III (1600 IU PMSG). Ewes were naturally mated and followed by laparotomy 6 days later. After laparotomy, ewes were injected with a prostaglandin analogue (PGF) and serum samples were obtained prior to surgery and then for 25 days to measure progesterone (P4) by radioimmunoassay. There were no differences among groups neither for estrous incidence (Group I: 83.3%; Group II: 83.3%; Group III: 100%), nor for the time interval to estrous onset (Group I: 26.4±2.4 h; Group II: 28.8±2.9 h; Group III: 24.0±3.8 h). Group I had more corpora lutea than Group II (14.2±1.2 and 6.2±0.8; P<0.05), and Group III was intermediate (11.0±3.0). There was a low incidence of persistent follicles in all treatments (Group I: 0.5±0.5; Group II: 0.6±0.4; Group III: 1.8±1.2). Number of collected ova were 9.0±2.6, 3.8±0.6 and 6.5±0.9 for Groups I, II and III, respectively. Significant differences in number of ova were detected between Groups I and II. Unfertilized ova did not differ among groups (Group I: 3.5±1.0; Group II: 2.8±0.8; Group III: 5.2±1.4; P>0.05). Embryos and high viability embryos were higher (P<0.05) in Group I (5.2±1.9 and 4.8±2.0) than in Group II (1.0±0.5 and 1.0±0.5) or Group III (1.2±0.6 and 1.0±0.5). Total plasma progesterone (P4) and P4 per corpus luteum before PGF administration did not vary (P>0.05) among groups (Group I: 71.0±14.7 and 4.9±0.7 nmol/l; Group II: 50.6±13.3 and 7.9±1.6 nmol/l; Group III: 90.4±42.6 and 6.8±1.8 nmol/l). There was a significant and positive correlation between P4 before PGF administration and number of corpora lutea (r=0.76). No significant differences were detected among groups for: interval PGF to P4 <3.18 nmol/l (Group I: 2.7±0.3 days; Group II: 1.8±0.6 days; Group III: 2.2±0.5 days), cycle length (Group I: 18.3±1.4 days; Group II: 17.9±0.5 days; Group III: 16.8±0.9 days), duration of P4 levels <3.18 nmol/l (Group I: 11.3±1.9 days; Group II: 7.1±1.0 days; Group III: 7.2±2.4 days), duration of P4 levels ≥3.18 nmol/l (Group I: 7.0±1.3 days; Group II: 10.8±0.8 days; Group III: 9.5±1.7 days) and peak of P4 (Group I: 7.4±0.4 nmol/l; Group II: 10.8±1.6 nmol/l; Group III: 9.2±1.9 nmol/l). It was concluded that PMSG–FSH-p treatment was more efficient than PMSG alone for superovulation and embryo production in ewes while P4 profiles were similar among groups.     

Effect of partial replacement of concentrates with jackfruit (Artocarpus heterophyllus) leaves on growth performance of kids grazing on native pasture of Tripura,India

Fifteen Black Bengal kids of about 3 months of age and body weight ranging from 3.8 to 4.9 kg were randomly distributed into three groups of five. Kids grazed native pasture 8 h/d. The kids in group I received supplementary concentrate (maize 35%, mustard cake 32%, rice bran 30%, mineral mixture 2% and common salt 1%) at approximately 2% of BW. However, 25 and 50% of the concentrate was replaced with jackfruit leaves for groups II and III, respectively. Total dry matter intake (DMI) was significantly higher in groups II and III than for group I due to greater forage consumption. Digestibility of CP (P < 0.05) decreased and that of NDF increased (P < 0.01) with increasing level of jackfruit leaves in the diet. Digestibility of ADF (P < 0.01), hemi cellulose (P < 0.05) and cellulose (P < 0.01) was higher in groups II and III in comparison to group I. Ruminal pH and TVFA concentration were not significantly different among the groups; however, rumen ammonia-N concentration decreased (P < 0.01) with increased level of jackfruit leaves in the diet. Similarly, plasma urea nitrogen and blood glucose levels were also reduced (P < 0.05) with increasing level of jackfruit leaves in the diet Average daily gain (ADG) was 47.33, 45.11 and 35.56 g/d in groups I, II and III, respectively. ADG and DMI/kg gain were not adversely affected when the level of replacement was restricted to 25%; however, at the 50% of replacement both parameters were adversely affected (P < 0.05). From the results of this experiment, it was concluded that jackfruit leaves might replace 25% of the supplemental concentrate for growing kids grazing in native pasture of northeast India.     

Chemical access to the mononuclear Mn(III) [(mL)Mn(OMe)]+ complex (mLH = N,N′-bis-(2-pyridylmethyl)-N-(2-hydroxybenzyl)-N′-methyl-ethane-1,2-diamine) and electrochemical oxidation to the Mn(IV) [(mL)Mn(OMe)]2+ species

Chemical oxidation in acetonitrile of the previously reported phenolato-bridged binuclear Mn(II) complex [(mL)MnMn(mL)]²⁺ (1), where mLH is pentadentate N,N′-bis-(2-pyridylmethyl)-N-(2-hydroxybenzyl)-N′-methyl-ethane-1,2-diamine ligand [C. Hureau, et al., Chem. Eur. J. 2004, 10, 1998–2010] using iodosylbenzene PhIO (dissolved in methanol) is described. The addition of one to four equivalents of PhIO per Mn ion leads to the transient formation of the mono-μ-oxo binuclear Mn₂(III,III) complex [(mL)Mn(μ-O)Mn(mL)]²⁺ (2), previously studied. After addition of five equivalents of PhIO per Mn ion, the mononuclear Mn(III) species [(mL)Mn(OMe)]⁺ (3) is quantitatively generated. The UV–Vis spectrum of 3 displays a broad band at 456 nm (ε = 1000 L mol⁻¹ cm⁻¹) attributed to phenolato to Mn(III) charge transfer transition. Complex 3 exhibits a reversible oxidation wave at E^1/2 = 0.68 V versus SCE, and the mononuclear Mn(IV) complex [(mL)Mn(OMe)]²⁺ (3^ox) can thus be generated by exhaustive electrolysis at 1.0 V versus SCE. The 9.4 GHz EPR spectrum of complex 3^ox shows a strong transition near g = 4 consistent with a rhombically distorted S = 3/2 system with a zero-field splitting dominating the Zeeman effect. UV–Vis spectrum displays a large phenolato to Mn(IV) charge transfer transition at 670 nm (ε = 2450 L mol⁻¹ cm⁻¹).     

Saponins from the roots of Chiococca alba and their in vitro anti-inflammatory activity

Five triterpenoidal saponins were isolated from the roots of Chiococca alba (L.) Hitchc. (Rubiaceae). Two of the saponins, chiococcasaponin III (3-O-β-d-glucopyranurosyl-3β-hydroxyolean-12,15-dien-28-oic acid 28-O-β-d-xylopyranosyl (1 → 4)-α-l-rhamnopyranosyl (1 → 2)-α-l-arabinopyranosyl ester) and chiococcasaponin IV (3-O-β-d-glucopyranurosyl-3β-hydroxyolean-12,15-dien-28-oic acid 28-O-α-l-rhamnopyranosyl (1 → 2)-α-l-arabinopyranosyl ester) were new and their structures were elucidated on the basis of extensive application of NMR techniques and high resolution electrospray mass spectrometry together with acid hydrolysis product analysis. As part of our investigations on the chemical profile and pharmacological activity of the roots of C. alba, we report the results of the evaluation of the activity of the saponin fractions against in vitro lipopolysaccharide-induced inflammation. The results found, strongly support the fractions I, III and IV as having anti-inflammatory properties.     

Analysis of a volumetric-modulated arc therapy (VMAT) single phase prostate template as a class solution

AimTo assess a class solution template for volumetric-modulated arc therapy (VMAT) for prostate cancer using plan analysis software.BackgroundVMAT is a development of intensity-modulated radiotherapy (IMRT) with potential advantages for the delivery of radiotherapy (RT) in prostate cancer. Class solutions are increasingly used for facilitating RT planning. Plan analysis software provides an objective tool for evaluating class solutions.Materials and methodsThe class solution for VMAT was based on the current static field IMRT template. The plans of 77 prostate cancer patients were evaluated using a set of in-house plan quality metrics (scores) (PlanIQ™, Sun Nuclear Corporation). The metrics compared the class solution for VMAT planning with the IMRT template and the delivered clinical plan (CP). Eight metrics were associated with target coverage and ten with organs-at-risk (OAR). Individual metrics were summed and the combined scores were subjected to non-parametric analysis. The low-dose wash for both static IMRT and VMAT plans were evaluated using 40 Gy and 25 Gy isodose volumes.ResultsVMAT plans were of equal or better quality than the IMRT template and CP for target coverage (combined score) and OAR combined score. The 40 Gy isodose volume was marginally higher with VMAT than IMRT (4.9%) but lower than CP (−6.6%)(P = 0.0074). The 25 Gy volume was significantly lower with VMAT than both IMRT (−32.7%) and CP (−34.4%)(P < 0.00001).ConclusionsAutomated VMAT planning for prostate cancer is feasible and the plans are equal to or better than the current IMRT class solution and the delivered clinical plan.     

Comparison of two type IV hyperthermophilic adenylyl cyclases characterizations from the archaeon Pyrococcus furiosus

In this paper, two genes that encoded two soluble type IV adenylyl cyclases (AC) from the hyperthermophilic archaeon Pyrococcus furiosus (PFAC I and PFAC II) were cloned and expressed in Escherichia coli (E. coli) BL21 (DE3). Amino acid sequence analysis of the two enzymes showed 29% homology. PFAC I and PFAC II were both Mn²⁺ activated enzyme. They were purified by His-trap chromatography and had a specific activity of 3.1 × 10³ U/mg at pH 10.0, 95 °C (PFAC I) and 2.0 × 10³ U/mg at pH 11.0, 95 °C (PFAC II), respectively. The K_m and k_cat of PFAC I was 1.38 mM and 1.11 s⁻¹. The K_m and k_cat of PFAC II was 1.44 mM and 0.80 s⁻¹. The thermostability of PFAC I and PFAC II were higher than the soluble type IV adenylyl cyclases from Yersinia pestis (YpAC-IV). All of the properties suggested that these two adenylyl cyclases may be useful for the industrial producing of cyclic adenosine 3′,5′-monophosphate (cAMP).