Cysteine proteases XCP1 and XCP2 aid micro-autolysis within the intact central vacuole during xylogenesis in Arabidopsis roots

Establishing the mechanisms regulating the autolysis of xylem tracheary elements (TEs) is important for understanding this programmed cell death process. These data demonstrate that two paralogous Arabidopsis thaliana proteases, XYLEM CYSTEINE PROTEASE1 (XCP1) and XCP2, participated in micro-autolysis within the intact central vacuole before mega-autolysis was initiated by tonoplast implosion. The data acquisition was aided by the predictable pattern of seedling root xylogenesis, the availability of single and double total knock-out T-DNA lines, anti-sera that recognized XCP1 and XCP2, and the microwave-assisted processing of whole seedlings prior to immunolabeling and observation in the transmission electron microscope. During secondary wall thickening, XCP1 and XCP2 (in wild type), XCP1 (in xcp2 seedlings) or XCP2 (in xcp1 seedlings) were imported into the TE central vacuole. Both XCP1 and XCP2 heavily labeled dense aggregates of material within the vacuole. However, because of XCP1 deficiency in xcp1 and xcp1 xcp2 TEs, non-degraded cellular remnants first accumulated in the vacuole and then persisted in the TE lumen (longer than in the wild type) after the final mega-autolysis was otherwise complete. This delayed TE clearing phenotype in xcp1 was rescued by complementation with wild-type XCP1. Although TEs in the xcp2 single knock-out cleared comparably with wild type, the non-degraded remnants in xcp1 xcp2 TEs were more densely packed than in xcp1 TEs. Therefore, XCP2 has a minor but distinct role in micro-autolysis. After tonoplast implosion, XCP1 and XCP2 remained associated with disintegrating cellular material as mega-autolysis, aided by additional lytic enzymes, destroyed the bulk of the cellular contents.     

Synthesis and antiprotozoal activity of novel bis-benzamidino imidazo[1,2-a]pyridines and 5,6,7,8-tetrahydro-imidazo[1,2-a]pyridines

The key dinitrile intermediates 4a-d were synthesized by reaction of phenacyl bromide 1 and the appropriate 2-amino-5-bromopyridines to yield 3a-d. Suzuki coupling of 3a-d with 4-cyanophenylboronic acid yielded the 2,6-bis(4-cyanophenyl)-imidazo[1,2-a]pyridine derivatives 4a-d. The bis-amidoximes 5a-d, obtained from 4a-d by the action of hydroxylamine, were converted to the bis-O-acetoxyamidoximes which on catalytic hydrogenation in a mixture of ethanol/ethyl acetate gave the acetate salts of 2,6-bis[4-(amidinophenyl)]-imidazo[1,2-a]pyridines 7a-d. In contrast, catalytic hydrogenation of the bis-O-acetoxyamidoxime of 5a in glacial acetic acid gave the saturated analogue 2,6-bis[4-(amidinophenyl)]-5,6,7,8-tetrahydro-imidazo[1,2-a]pyridine 8. O-Methylation of the amidoximes 5a-d gave the N-methoxyamidines 6a-d. The diamidines showed strong DNA binding affinity, were very active in vitro against T. b. r. exhibiting IC(50) values between 7 and 38nM, but were less effective against P. f. with IC(50) values between 23 and 92nM. Two of the diamidines 7c and 7d were slightly more active than furamidine but less active than azafuramidine in the T. b. r. STIB900 mouse model. Only one prodrug 6b showed moderate activity in the same mouse model.     

Sex determination using the femur in an ancient Japanese population

Determination of sex from the femur measurements has been attempted in several populations and various studies have demonstrated the importance of population specific standards in the metric assessment of sex. The present study attempts to establish metric standards for sex determination by using femur measurements for ancient Japanese populations. Osteometric data were obtained from 151 adult skeletal remains from Jomon period, Yoshigo human skeletal collection. Eight femur measurements were taken and the data were analyzed by discriminant analysis using SPSS version 10.0. For the univariate discriminant function derived, precision of sex determination was 93% with the condyle breadth. Prediction values showed that sex differentiation could be done by femur measurements with reliability between 66.9 and 100%, with values for males higher than for females. It is suggested that discriminant formulas developed by femur measurements in this study, can be used for sex determination accurately on fragmentary skeletal remains in ancient Japanese populations.     

<Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> and Its Probiotic and Food Potentialities

The number of studies claiming probiotic health effects of Lactobacillus plantarum is escalating. Lb. plantarum is a lactic acid bacterium found in diverse ecological niches, highlighting its particular capabilities of adaptation and genome plasticity. Another function that needs to be underlined is the capabilities of Lb. plantarum to produce diverse and potent bacteriocins, which are antimicrobial peptides with possible applications as food preservative or antibiotic complementary agents. Taken together, all these characteristics design Lb. plantarum as a genuine model for academic research and viable biological agent with promising applications. The present review aims at shedding light on the safety of Lb. plantarum and run through the main studies underpinning its beneficial claims. The mechanisms explaining probiotic-related features are discussed.     

Design,synthesis and anticancer evaluation of novel pyrazole,pyrazolo[3,4-d]pyrimidine and their glycoside derivatives

The chalcone derivatives 3a,b were cyclized upon reaction with thiourea to give the pyrazolo[3,4-d]pyrimidine derivatives 5a,b. Condensation of 5a,b and their hydrazide derivatives 8a,b with cyclic and acyclic glucose gave the condensed S- and N-glycosides 7a,b and 9a,b, respectively. Reaction of 3b with ethyl cyanoacetate followed by reaction with cyclic glucose afforded a mixture of the O- and/or N-glycoside isomers 12 and 13, respectively. The pyrazolo[3,4-c]pyrazole derivative 14 was also obtained from the reaction of 3b with hydrazine hydrate. A number of the synthesized compounds were screened for their antitumor activity against three different tumor cell lines HEPG2 (liver), HCT116 (colon) and MCF-7 (breast) with a docking study against CDK2.     

Bioavailability of transgenic microRNAs in genetically modified plants

Background

Transgenic expression of small RNAs is a prevalent approach in agrobiotechnology for the global enhancement of plant foods. Meanwhile, emerging studies have, on the one hand, emphasized the potential of transgenic microRNAs (miRNAs) as novel dietary therapeutics and, on the other, suggested potential food safety issues if harmful miRNAs are absorbed and bioactive. For these reasons, it is necessary to evaluate the bioavailability of transgenic miRNAs in genetically modified crops.

Results

As a pilot study, two transgenic Arabidopsis lines ectopically expressing unique miRNAs were compared and contrasted with the plant bioavailable small RNA MIR2911 for digestive stability and serum bioavailability. The expression levels of these transgenic miRNAs in Arabidopsis were found to be comparable to that of MIR2911 in fresh tissues. Assays of digestive stability in vitro and in vivo suggested the transgenic miRNAs and MIR2911 had comparable resistance to degradation. Healthy mice consuming diets rich in Arabidopsis lines expressing these miRNAs displayed MIR2911 in the bloodstream but no detectable levels of the transgenic miRNAs.

Conclusions

These preliminary results imply digestive stability and high expression levels of miRNAs in plants do not readily equate to bioavailability. This initial work suggests novel engineering strategies be employed to enhance miRNA bioavailability when attempting to use transgenic foods as a delivery platform.

     

The Effect of Molecular Adsorption on Electro-Optical Properties of Graphene-Based Sensors

Extraordinary electrical and optical features of graphene-based materials attract researchers to improve sensing center of different sensors using them. In this research, the effects of sensing molecules on electro-optical features of graphene-based sensors are modeled. The adsorption effect on the Hamiltonian of the system based on tight-binding model is explored, and also the system band structure is investigated analytically. Then, refractive index deviations based on band gap variations are discovered which are used in response modeling of a graphene-based surface plasmon resonance (SPR) sensor.

     

Application of BATMAN and BAYESIL for quantitative <Superscript>1</Superscript>H-NMR based metabolomics of urine: discriminant analysis of lean,obese, and obese-diabetic rats

Introduction

BATMAN and BAYESIL are software tools, which can provide a solution for automated metabolite quantifications based on the proton nuclear magnetic resonance (¹H-NMR) spectral data of bio-fluids. However, their specific application for the quantitative ¹H-NMR based metabolomics of urine has not been investigated.

Objectives

The aim of this study is to evaluate the performance of BATMAN and BAYESIL in the quantitative metabolite analysis of urine based on its ¹H-NMR spectra.

Methods

BATMAN and BAYESIL were used for automated metabolite quantification based on the ¹H-NMR spectra of the urine from the lean, obese and obese-diabetic rat groups. PLS-DA model was used to discriminate the three different groups based on the results from the quantifications.

Results

BATMAN was found to be superior to BAYESIL in identifying and quantifying the metabolites in the urine samples, owing to its flexibility that allows users to define and adjust the relevant signals of the pure standard metabolites in the database in order to fit the signals in the samples, a necessary step since variations and peak shift are natural in most ¹H-NMR spectra. The results of BATMAN also agreed well with that of the manual deconvolution method, which indicated the higher accuracy in metabolite quantification, despite the need of pre-processing and longer processing time than BAYESIL. However, in the case where the problems in baseline correction and peak shift of ¹H-NMR spectra are absent, the use of BAYESIL is more advantageous. Application of quantitative ¹H-NMR based metabolomics of the urine showed that PLS-DA model derived from BATMAN could satisfactorily discriminate the lean, obese, and obese-diabetic rat groups.

Conclusion

Both BATMAN and BAYESIL are useful for the quantitative automation of urine metabolites based on its ¹H-NMR spectra. The results from BATMAN method is superior to BAYESIL but require expertise in spectroscopy and longer computer time. Both methods help in simplifying the interpretation of metabolite status in the VIP analysis.

     

Polystoma macrocnemis n. sp. (Monogenea: Polystomatidae) from the Iranian longlegged wood frog Rana macrocnemis (Ranidae) in Turkey

Polystoma macrocnemis n. sp. (Polystomatidae) is described from the urinary bladder of Rana macrocnemis (Ranidae) in Turkey. Its general morphology is similar to that of other members of the genus, but it is distinguished from closely related species by numerical parameters of the hamuli and also by its host species and geographical range.