Correction to: Identification of the meiotic toolkit in diatoms and exploration of meiosis-specific SPO11 and RAD51 homologs in the sexual species Pseudo-nitzschia multistriata and Seminavis robusta.

Following the publication of this article [1], the authors reported that the link to Additional file 11 linked to the wrong set of data. The correct supplementary data is provided in this Correction article (Additional file 11).

In vitro Protective Effect of Rutin and Quercetin against Radiation-induced Genetic Damage in Human Lymphocytes.

PURPOSE OF THE STUDY: Rutin (RUT) and quercetin (QRT) which are dietary compounds were investigated for their ability to protect against ionizing radiation (IR)-induced genotoxicity in human lymphocytes. MATERIALS AND METHODS: The radiation antagonistic potential of RUT and QRT was assessed by alkaline comet and cytokinesis-block micronucleus (CBMN) assay. RESULTS: Treatment of lymphocytes with RUT and QRT (25 µg/ml) prior exposure to 2 Gy gamma radiation resulted in a significant reduction of frequency of micronuclei as compared to the control set of cells evaluated by CBMN assay. Similarly, treatment of lymphocytes with RUT and QRT before radiation exposure showed significant decrease in the DNA damage as assessed by comet parameters, such as percent tail DNA and olive tail moment. CONCLUSION: The study demonstrates the protective effect of RUT and QRT against IR-induced DNA damage in human lymphocytes, which may be partly attributed to scavenging of IR-induced free radicals and also by the inhibition of IR-induced oxidative stress.

Finding a partner in the ocean: molecular and evolutionary bases of the response to sexual cues in a planktonic diatom.

Microalgae play a major role as primary producers in aquatic ecosystems. Cell signalling regulates their interactions with the environment and other organisms, yet this process in phytoplankton is poorly defined. Using the marine planktonic diatom Pseudo-nitzschia multistriata, we investigated the cell response to cues released during sexual reproduction, an event that demands strong regulatory mechanisms and impacts on population dynamics. We sequenced the genome of P. multistriata and performed phylogenomic and transcriptomic analyses, which allowed the definition of gene gains and losses, horizontal gene transfers, conservation and evolutionary rate of sex-related genes. We also identified a small number of conserved noncoding elements. Sexual reproduction impacted on cell cycle progression and induced an asymmetric response of the opposite mating types. G protein-coupled receptors and cyclic guanosine monophosphate (cGMP) are implicated in the response to sexual cues, which overall entails a modulation of cell cycle, meiosis-related and nutrient transporter genes, suggesting a fine control of nutrient uptake even under nutrient-replete conditions. The controllable life cycle and the genome sequence of P. multistriata allow the reconstruction of changes occurring in diatoms in a key phase of their life cycle, providing hints on the evolution and putative function of their genes and empowering studies on sexual reproduction.

Effect of dietary supplementation of garlic, ginger and their combination on feed intake, growth performance and economics in commercial broilers.

AIM: The present study was carried out to evaluate the effect of supplementation of garlic, ginger and their combination in the diets of broiler chickens and assessment in terms of feed intake, growth performance and economics of feeding. MATERIALS AND METHODS: A total of 240 1-day-old Cobb-400 broiler chicks were randomly assigned to four dietary treatments each with three replicates of 20 chicks per replicate (n=60). Four experimental diets were formulated in such a way that control diet (T1) contained neither ginger nor garlic. While, birds in group T2 and T3 were fed with diets containing 1% garlic and ginger, respectively. Diet 4 (T4 group) contained a combination of 1% of garlic and ginger. The feeding experiment was carried out for 42 days, and different parameters evaluated includes feed intake, weight gain, feed conversion ratio, gut morphometry, and economics of feeding in terms of return over feed cost (ROFC) and European Performance Efficiency Index. RESULTS: Feed intake of experimental birds in ginger and mixture of garlic and ginger supplemented groups, i.e., T3 and T4 groups have significantly (p<0.05) higher feed intake as compared to control. While, feeding of garlic have non-significant effect on feed intake as compared to other groups. A body weight gain (g/bird) was found to be significantly (p<0.05) higher in garlic (T2 group) and ginger (T3 group) supplemented group as compare to control and garlic and ginger mixture supplemented group (T4 group). Feed conversion ratio was significantly (p<0.05) lower in ginger (T3 group) supplemented group as compare to other groups. Mean villi length, villi width and cryptal depth were significantly (p<0.05) higher in T3 group than rest of all three groups, indicating increased absorptive surface area. ROFC was significantly (p<0.05) lower in T3 and T4 groups as compare to control. However, it was not significantly different between control and T2 group. CONCLUSION: On the basis of the results of the study, it is concluded that supplementation of garlic improves the performance of broilers when added at the rate of 1% of broiler ration and can be a viable alternative to antibiotic growth promoter in the feeding of broiler chicken.

Identification of the meiotic toolkit in diatoms and exploration of meiosis-specific SPO11 and RAD51 homologs in the sexual species Pseudo-nitzschia multistriata and Seminavis robusta.

BACKGROUND: Sexual reproduction is an obligate phase in the life cycle of most eukaryotes. Meiosis varies among organisms, which is reflected by the variability of the gene set associated to the process. Diatoms are unicellular organisms that belong to the stramenopile clade and have unique life cycles that can include a sexual phase. RESULTS: The exploration of five diatom genomes and one diatom transcriptome led to the identification of 42 genes potentially involved in meiosis. While these include the majority of known meiosis-related genes, several meiosis-specific genes, including DMC1, could not be identified. Furthermore, phylogenetic analyses supported gene identification and revealed ancestral loss and recent expansion in the RAD51 family in diatoms. The two sexual species Pseudo-nitzschia multistriata and Seminavis robusta were used to explore the expression of meiosis-related genes: RAD21, SPO11-2, RAD51-A, RAD51-B and RAD51-C were upregulated during meiosis, whereas other paralogs in these families showed no differential expression patterns, suggesting that they may play a role during vegetative divisions. An almost identical toolkit is shared among Pseudo-nitzschia multiseries and Fragilariopsis cylindrus, as well as two species for which sex has not been observed, Phaeodactylum tricornutum and Thalassiosira pseudonana, suggesting that these two may retain a facultative sexual phase. CONCLUSIONS: Our results reveal the conserved meiotic toolkit in six diatom species and indicate that Stramenopiles share major modifications of canonical meiosis processes ancestral to eukaryotes, with important divergences in each Kingdom.

Transcriptome sequencing of three Pseudo-nitzschia species reveals comparable gene sets and the presence of Nitric Oxide Synthase genes in diatoms.

Diatoms are among the most diverse eukaryotic microorganisms on Earth, they are responsible for a large fraction of primary production in the oceans and can be found in different habitats. Pseudo-nitzschia are marine planktonic diatoms responsible for blooms in coastal and oceanic waters. We analyzed the transcriptome of three species, Pseudo-nitzschia arenysensis, Pseudo-nitzschia delicatissima and Pseudo-nitzschia multistriata, with different levels of genetic relatedness. These species have a worldwide distribution and the last one produces the neurotoxin domoic acid. We were able to annotate about 80% of the sequences in each transcriptome and the analysis of the relative functional annotations allowed comparison of the main metabolic pathways, pathways involved in the biosynthesis of isoprenoids (MAV and MEP pathways), and pathways putatively involved in domoic acid synthesis. The search for homologous transcripts among the target species and other congeneric species resulted in the discovery of a sequence annotated as Nitric Oxide Synthase (NOS), found uniquely in Pseudo-nitzschia multistriata. The predicted protein product contained all the domains of the canonical metazoan sequence. Putative NOS sequences were found in other available diatom datasets, supporting a role for nitric oxide as signaling molecule in this group of microalgae.

Establishment of Genetic Transformation in the Sexually Reproducing Diatoms Pseudo-nitzschia multistriata and Pseudo-nitzschia arenysensis and Inheritance of the Transgene.

We report the genetic transformation of the planktonic diatoms Pseudo-nitzschia arenysensis and Pseudo-nitzschia multistriata, members of the widely distributed and ecologically important genus Pseudo-nitzschia. P. arenysensis and P. multistriata present the classical size reduction/restitution life cycle and can reproduce sexually. Genetic transformation was achieved with the biolistic method, using the H4 gene promoter from P. multistriata to drive expression of exogenous genes. The transformation was first optimized introducing the Sh ble gene to confer resistance to the antibiotic zeocin. Integration of the transgene was confirmed by PCR and Southern blot analyses. Subsequently, we simultaneously transformed in P. arenysensis two plasmids, one encoding the ß-glucuronidase (GUS) gene together with the plasmid carrying the Sh ble resistance gene, demonstrating the possibility of co-transformation. By transforming a gene encoding a fusion between the histone H4 and the green fluorescent protein (GFP), we demonstrated that fluorescent tagging is possible and that studies for protein localization are feasible. Importantly, we crossed P. arenysensis- and P. multistriata-transformed strains with a wild-type strain of opposite mating type and demonstrated that the transgene can be inherited in the F1 generation. The possibility to transform two diatom species for which genetic crosses are possible opens the way to a number of new approaches, including classical loss of function screens and the possibility to obtain different combinations of double transformants.

Antigenotoxic potential of rutin and quercetin in Swiss mice exposed to gamma radiation.

BACKGROUND: Ionizing radiation induces a variety of genetic damages through the formation free radicals such as reactive oxygen species (ROS). Appropriate antioxidant intervention may inhibit or reduce free radical toxicity and thus offer protection against radiation. Rutin (RUT) and quercetin (QRT) are flavonoids known to be potent dietary antioxidants. METHODS: The present study tested the antigenotoxic effect of RUT and QRT in vivo against radiation- induced chromosomal damage. Swiss albino mice were administered orally with RUT and QRT (10 and 20 mg/kg b.wt.) once daily for five consecutive days. One hour after the last administration of RUT and QRT on the fifth day, the animals were whole body exposed to 3 Gy gamma radiation. The anti-genotoxic potential was assessed in terms of chromosomal aberrations, micronucleus test, and alkaline comet assay. RESULTS: Significant decline in dicentric formation was observed in RUT and QRT treated group. Further, the antigenotoxic potential of RUT and QRT caused a significant (p < 0.001) reduction in micronucleated polychromatic, normochromatic erythrocytes; increased PCE/NCE ratio was observed in the RUT and QRT treated group. Administration of RUT and QRT before irradiation resulted in a significant (p < 0.01) decrease in the DNA damage at the post-irradiation time when compared with irradiation alone group. CONCLUSIONS: Present findings demonstrate the potential of RUT and QRT in mitigating radiation-induced mortality and cytogenetic damage, which may be attributed to scavenging of radiation-induced free radicals.

Antioxidative and radioprotective potential of rutin and quercetin in Swiss albino mice exposed to gamma radiation.

The radioprotective potential of bioflavonoid, rutin (RUT) and quercetin (QRT) was investigated in Swiss albino mice exposed to gamma radiation. The radioprotective potential of RUT and QRT was assessed in pre-treatment group of mice followed on radiation-induced changes in glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation (LPO) levels were also analyzed. Elevation in the GSH, GST, SOD, CAT, and decreased LPO levels were observed in RUT and QRT pretreated group when compared to the irradiated animals. Furthermore, it was observed that RUT and QRT treatment was found to inhibit various free radicals generated in vitro, viz., 2,2-diphenyl-1-picrylhydrazyl(DPPH), O2, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS)(+), and OH in a concentration-dependent manner. This study clearly demonstrates the free radical scavenging action of RUT and QRT, indicating that it may have its potential as a radioprotective agent. Furthermore, the presence of a phenolic group in RUT and QRT is known to contribute to scavenging the radiation-induced free radicals and inhibition of oxidative stress. Present findings demonstrate the potential of RUT and QRT in mitigating radiation-induced oxidative stress, which may be attributed to the inhibition of radiation-induced decline in the endogenous antioxidant levels and scavenging of radiation-induced free radicals.

Evaluation of biofield treatment dose and distance in a model of cancer cell death.

OBJECTIVE: This study assessed the potential influence of biofield treatment on cultured human cancer cells and whether such influence was affected by varying the duration of the treatment (dose) or the distance between the biofield practitioner and the target cells. DESIGN: Biofield treatment dosage was assessed from a short distance (0.25 meters) in three independent experiments involving 1, 2, or 5 treatments, along with another set of three independent and comparable mock experiments. Biofield treatment distance was assessed at 0.25, 25, and ∼ 2000 meters involving two treatments in three independent experiments along with another set of three mock experiments. INTERVENTION: Biofield treatments were delivered by a highly acclaimed biofield practitioner with the intention of diminishing growth of the cells or inducing cancer-cell death. OUTCOME MEASURE: Cell viability was quantified 20 hours after treatments, using a spectrophotometric assay for live-cell counting. The dependent measure for each experiment was the log ratio of the cell viability values of treated samples (biofield or mock) over the values of untreated control samples. RESULTS: A trend of decreasing cell viability with increasing biofield dose was evident in the first set of experiments assessing dose-response; however, no such effect was evident in the second set of experiments evaluating biofield treatment distance. Mock experiments yielded relatively stable viability ratios in both sets of experiments. Linear regression analysis and hypothesis testing of the data taken as a whole did not yield statistical significance at p<0.05. CONCLUSIONS: These results represent the first indication of a biofield treatment dose-response in a controlled laboratory setting. The data are inconclusive because of the inability of reproduce the cellular response in a replicate experiment.

Simultaneous quantification of berberine and lysergol by HPLC-UV: evidence that lysergol enhances the oral bioavailability of berberine in rats.

A sensitive and simple HPLC method was developed for the simultaneous quantification of berberine and lysergol in rat plasma. The chromatographic separation was achieved on a C(18) column using isocratic elution with methanol-acetonitrile-0.1% ortho-phosphoric acid (25:20:55, v/v/v), pH adjusted to 6.5 with triethylamine and detected at a UV wavelength of 230 nm. The extraction of the berberine and lysergol from the rat plasma with methylene chloride resulted in their high recoveries (82.62 and 90.17%). HPLC calibration curves for both berberine and lysergol based on the extracts from the rat plasma were linear over a broad concentration range of 50-1000 ng/mL. The limit of quantification was 50 ng/mL. Intra- and inter-day precisions were <15% and accuracy was 87.12-92.55% for berberine and 87.01-92.26% for lysergol. Stability studies showed that berberine and lysergol were stable in rat plasma for short- and long-term period for sample preparation and analysis. The described method was successfully applied to study the pharmacokinetics of berberine as well as lysergol following oral administration in Sprague-Dawley rats. The results of the study inferred that lysergol improved the oral bioavailability of berberine.

Novel and validated titrimetric method for determination of selected angiotensin-II-receptor antagonists in pharmaceutical preparations and its comparison with UV spectrophotometric determination.

A novel and simple titrimetric method for determination of commonly used angiotensin-II-receptor antagonists (ARA-IIs) is developed and validated. The direct acid base titration of four ARA-IIs, namely eprosartan mesylate, irbesartan, telmisartan and valsartan, was carried out in the mixture of ethanol:water (1:1) as solvent using standardized sodium hydroxide aqueous solution as titrant, either visually using phenolphthalein as an indicator or potentiometrically using combined pH electrode. The method was found to be accurate and precise, having relative standard deviation of less than 2% for all ARA-IIs studied. Also, it was shown that the method could be successfully applied to the assay of commercial pharmaceuticals containing the above-mentioned ARA-IIs. The validity of the method was tested by the recovery studies of standard addition to pharmaceuticals and the results were found to be satisfactory. Results obtained by this method were found to be in good agreement with those obtained by UV spectrophotometric method. For UV spectrophotometric analysis ethanol was used as a solvent and wavelength of 233 nm, 246 nm, 296 nm, and 250 nm was selected for determination of eprosartan mesylate, irbesartan, telmisartan, and valsartan respectively. The proposed titrimetric method is simple, rapid, convenient and sufficiently precise for quality control purposes.

Radiomodulatory role of Rutin and Quercetin in Swiss Albino mice exposed to the whole body gamma radiation.

CONTEXT: Radiation therapy is the prime treatment modality against various cancers. However, its use is limited due to the effects of radiation on normal tissues. AIMS: In view of this, present study was carried out to evaluate the radioprotective potential of Rutin (RUT) and Quercetin (QRT) in Swiss Albino mice exposed to the whole body gamma radiation. To gain insight into the mechanism of action, RUT and QRT were tested for its antioxidant levels in mice. SETTINGS AND DESIGNS: Optimum protective dose of RUT and QRT against radiation induced animal mortality was selected by administration of various doses of the RUT and QRT before 10 Gy gamma irradiation. MATERIALS AND METHODS: Swiss Albino mice were used for the assessment of radiation induced sickness along with the survival analysis and anti-oxidative properties of RUT and QRT. STATISTICAL ANALYSIS USED: Survival studies were determined using the Kaplan-Meier survival curves. RESULTS: The maximum survival was observed with 10 mg/kg. b. wt. and 20 mg/kg. b. wt. of RUT and QRT respectively, this dose was considered as an optimal dose for radioprotection. Treatment of mice with RUT and QRT before irradiation delayed the onset of mortality as compared with the untreated irradiated controls. The oral administration of RUT and QRT resulted in an increase in the radiation tolerance and the dose reduction factor was found to be 1.15 and 1.11 respectively. RUT and QRT pre-treatment significantly (P < 0.01) elevated levels of reduced glutathione, glutathione-S-transferase, catalase, Superoxide dismutase, and a decreased lipid peroxidation in mouse liver homogenate at 24 h after exposure to 4.5 Gy. CONCLUSIONS: Present findings demonstrate the potential of RUT and QRT in mitigating radiation-induced mortality, which may be attributed to the elevation in the antioxidant status, anti-lipid peroxidative potential.

Statistical optimization of floating pulsatile drug delivery system for chronotherapy of hypertension.

INTRODUCTION: A pulsatile drug delivery system is characterized by a lag time that is an interval of no drug release followed by rapid drug release. The purpose of this work was to develop hollow calcium alginate beads for floating pulsatile release of valsartan intended for chronopharmacotherapy. Floating pulsatile concept was applied to increase the gastric residence of the dosage form having lag phase followed by a burst release. MATERIALS AND METHODS: To overcome the limitations of various approaches for imparting buoyancy, hollow/porous beads were prepared by simple process of acid-base reaction during ionotropic crosslinking by low viscosity sodium alginate and calcium chloride as a crosslinking agent. In this study, investigation of the functionality of the sodium alginate to predict lag time and drug release was statistically analyzed using the response surface methodology (RSM). RSM was employed for designing of the experiment, generation of mathematical models and optimization study. The chosen independent variables, i.e. sodium alginate and potassium bicarbonate were optimized with a 3(2) full factorial design. Floating time and cumulative percentage drug release in 6 h were selected as responses. RESULTS: Results revealed that both the independent variables are significant factors affecting drug release profile. A second-order polynomial equation fitted to the data was used to predict the responses in the optimal region. The optimized formulation prepared according to computer-determined levels provided a release profile, which was close to the predicted values. The floating beads obtained were porous (21-28% porosity), hollow with bulk density <1 and had Ft(70) of 2-11 h. The floating beads provided expected two-phase release pattern with initial lag time during floating in acidic medium followed by rapid pulse release in phosphate buffer. CONCLUSION: The proposed mathematical model is found to be robust and accurate for optimization of time-lagged formulations for programmable pulsatile release of valsartan.

Variation in common lipid parameters in malaria infected patients.

The heart is remarkably resilient even in the face of heavy parasite sequestration and other vital organ dysfunction, and deaths from cardiac arrhythmias in severe malaria are rare. Malaria may prove fatal for patients with pre-existing cardiac failure due to valvular stenosis or myocardial disease. High grade fever, parasitaemia, and fluid overload can all contribute to the problem. Cardiac arrhythmias are very rarely observed in severe falciparum malaria. An attempt has been made to evaluate the risk factors for cardiovascular diseases in malaria infected patients. In the present study the levels of total cholesterol, low density lipoproteins, triglycerides were high and the levels of high density lipoproteins were low in malaria infected patients compared to controls. The markers of free radical induced injury i.e. malondialdehyde were high. The study therefore suggests the importance of assessing these markers of oxidative stress along with the other routine investigations in malaria infected patients for initiating therapy in addition to primary and secondary preventive measures to mitigate the devastating consequences hyperlipidemia in malaria infected patients leading to cardiovascular diseases.