Propensity-matched analysis does not support angiosome-guided revascularization of multilevel peripheral artery disease (PAD).

BACKGROUND: This retrospective comparative cohort study evaluated the clinical outcome of angiosome-guided endovascular arterial reconstructions in chronic limb-threatening ischemia (CLTI) due to multilevel peripheral artery disease (PAD). METHODS: Patients treated in an endovascular fashion for CLTI with tissue loss due to multilevel PAD were analyzed. Limbs were classified as having undergone either angiosome-guided (direct) revascularization (DR) or nonangiosomic (indirect) revascularization (IR). DR was defined as uninterrupted in-line flow to the affected angiosome, revascularization through the pedal arch was also considered direct. Groups were adjusted with propensity score (PS) matching and compared for amputation-free survival (AFS), freedom from major adverse limb events (MALE), and healing rate at 12 months. RESULTS: A total of 174 patients (81 men, mean age 70.0 ± 10.4 y) were included. PS matching produced two groups of 55 patients each: DR (24 men, mean age 71.7 ± 10.7 y) and IR (26 men, mean age 72.0 ± 9.4 y). The matched groups had no significant differences in baseline variables. At 12 months there were no significant differences in AFS (73.2% vs 71.6%; p = 0.841), freedom from MALE (71.7% vs 66.1%; p = 0.617), and healing rate (72.7% vs 72.0%; p = 1.000) between DR and IR, respectively. CONCLUSION: This study failed to support the use of angiosome concept in CLTI due to multilevel disease.

Popliteal Artery Angioplasty for Chronic Total Occlusions with versus without the Distal Landing Zone.

BACKGROUND: The purpose of this study was to evaluate the outcome of percutaneous transluminal angioplasty in patients with critical limb ischemia due to popliteal artery (PA) chronic total occlusions depending on the presence of a patent portion of the PA distal to the occlusive lesion-the distal landing zone (DLZ). MATERIALS AND METHODS: We retrospectively analyzed 80 patients with critical limb ischemia (all Rutherford class 5-6), who underwent percutaneous transluminal angioplasty with or without stenting for PA chronic total occlusions with no inflow disease. Baseline demographic and clinical variables, periprocedural outcome, 12-month overall survival, limb salvage, primary patency, freedom from target lesion revascularization (TLR), amputation-free survival, and freedom from major adverse limb events in DLZ versus no-DLZ lesions were assessed. RESULTS: Of all patients (43 men; mean age 70.2 y), 40 (50%) had DLZ in the PA, whereas another 40 (50%) did not (no-DLZ). Diabetes was significantly more common among DLZ patients and was found to be a risk factor for DLZ compared with no-DLZ lesions (HR 2.58; 95% CI 1.03-6.46; P = 0.04). Other demographic and clinical variables were similar between the groups. The stenting rate was 45.0% versus 42.5% in DLZ versus no-DLZ (P = 1.0). At 12 months, there was no significant difference in primary patency (64.7% vs. 51.6%; P = 0.156), overall survival (73.4% vs. 84.0%; P = 0.283), amputation-free survival (60.0% vs. 68.8%; P = 0.432), and limb salvage rate (93.6% vs. 82.2%; P = 0.126) between DLZ and no-DLZ groups, respectively. However, freedom from TLR (92.1% vs. 67.7%; P = 0.03) and major adverse limb events (80.1% vs. 41.8%; P = 0.003) was significantly higher in DLZ compared with no-DLZ lesions (92.1% vs. 67.7%; P = 0.03). CONCLUSIONS: Diabetes was found to be a significant risk factor for DLZ compared to no-DLZ lesions. Technical success and stenting rates were similar in DLZ versus no-DLZ patients. At 12 months, there was no significant difference in limb salvage, primary patency, and overall survival between the study groups. The DLZ lesions were associated with a significantly higher freedom from TLR than no-DLZ lesions.

Diabetes noninvasive diagnostics and monitoring through volatile biomarkers analysis in the exhaled breath using optical absorption spectroscopy.

The review is aimed on the analysis the abilities of noninvasive diagnostics and monitoring of diabetes mellitus (DM) and DM-associated complications through volatile molecular biomarkers detection in the exhaled breath. The specific biochemical reactions in the body of DM patients and their associations with volatile molecular biomarkers in the breath are considered. The applications of optical spectroscopy methods, including UV, IR, and terahertz spectroscopy for DM-associated volatile molecular biomarkers measurements, are described. The applications of similar technique combined with machine learning methods in DM diagnostics using the profile of DM-associated volatile molecular biomarkers in exhaled air or "pattern-recognition" approach are discussed.

Quantitative Assessment of Low-Dose Photodynamic Therapy Effects on Diabetic Wound Healing Using Raman Spectroscopy.

One of challenges that faces diabetes is the wound healing process. The delayed diabetic wound healing is caused by a complicated molecular mechanism involving numerous physiological variables. Low-dose photodynamic therapy (LDPDT) provides excellent results in rejuvenation and wound healing. In this study, the LDPDT effect on diabetic wounds in mice was studied using two photosensitizers, 5-aminolevulinic acid and methylene blue, and two laser dose expositions of 1 J/cm2 and 4 J/cm2 by Raman spectroscopy (RS). The latter was used as a noninvasive method, providing specific information about tissue state based on the fundamental vibrational modes of its molecular components. RS allows high spatial resolution acquisition of biochemical and structural information through the generation of point spectra or spectral images. An approach to in vivo quantitative assessment of diabetic wound healing state was developed. This approach is based on an application of the principal component analysis combined with the Mahalanobis metrics to skin Raman spectra, in particular, intensities of the amide I and CH2 bands.

A Criterion of Colorectal Cancer Diagnosis Using Exosome Fluorescence-Lifetime Imaging.

This study was aimed to investigate the applicability of the exosome fluorescence-lifetime imaging microscopy (FLIM) for colorectal cancer (CRC) diagnosis. Differential ultra-centrifugation was used to extract exosomes from the blood plasma of 11 patients with colon polyps (CPs) and 13 patients with CRC at the T2-4, N0-3, and M0-1 stages. Analysis was performed using a two-photon FLIM device. In total, 165 and 195 FLIM images were recorded for the CP and CCR patient groups, respectively. Two classes of exosomes differentiated by autofluorescence average lifetime tm were discovered in the samples. The first class of exosomes with tm = (0.21 ± 0.06) ns was mostly found in samples from CRC patients. The second class with tm = (0.43 ± 0.19) ns was mostly found in samples from CP patients. The relative number of "CRC-associated" exosomes Nch in the FLIM dataset was shown to be very small for the CP patient group and large for the CRC patient group. This difference was statistically significant. Therefore, the suggested CRS diagnostics criterion can be as follows. If Nch > 0.5, the probability of CRC is high. If Nch < 0.3, the probability of CRC is low.

IPD3 controls the formation of nitrogen-fixing symbiosomes in pea and Medicago Spp.

A successful nitrogen-fixing symbiosis requires the accommodation of rhizobial bacteria as new organelle-like structures, called symbiosomes, inside the cells of their legume hosts. Two legume mutants that are most strongly impaired in their ability to form symbiosomes are sym1/TE7 in Medicago truncatula and sym33 in Pisum sativum. We have cloned both MtSYM1 and PsSYM33 and show that both encode the recently identified interacting protein of DMI3 (IPD3), an ortholog of Lotus japonicus (Lotus) CYCLOPS. IPD3 and CYCLOPS were shown to interact with DMI3/CCaMK, which encodes a calcium- and calmodulin-dependent kinase that is an essential component of the common symbiotic signaling pathway for both rhizobial and mycorrhizal symbioses. Our data reveal a novel, key role for IPD3 in symbiosome formation and development. We show that MtIPD3 participates in but is not essential for infection thread formation and that MtIPD3 also affects DMI3-induced spontaneous nodule formation upstream of cytokinin signaling. Further, MtIPD3 appears to be required for the expression of a nodule-specific remorin, which controls proper infection thread growth and is essential for symbiosome formation.

Application of machine learning and laser optical-acoustic spectroscopy to study the profile of exhaled air volatile markers of acute myocardial infarction.

Conventional acute myocardial infarction (AMI) diagnosis is quite accurate and has proved its effectiveness. However, despite this, discovering more operative methods of this disease detection is underway. From this point of view, the application of exhaled air analysis for a similar diagnosis is valuable. The aim of the paper is to research effective machine learning algorithms for the predictive model for AMI diagnosis constructing, using exhaled air spectral data. The target group included 30 patients with primary myocardial infarction. The control group included 42 healthy volunteers. The 'LaserBreeze' laser gas analyzer (Special Technologies Ltd, Russia), based on the dual-channel resonant photoacoustic detector cell and optical parametric oscillator as the laser source, had been used. The pattern recognition approach was applied in the same manner for the set of extracted concentrations of AMI volatile markers and the set of absorption coefficients in a most informative spectral range 2.900 ± 0.125µm. The created predictive model based on the set of absorption coefficients provided 0.86 of the mean values of both the sensitivity and specificity when linear support vector machine (SVM) combined with principal component analysis was used. The created predictive model based on using six volatile AMI markers (C5H12, N2O, NO2, C2H4, CO, CO2) provided 0.82 and 0.93 of the mean values of the sensitivity and specificity, respectively, when linear SVM was used.

Symbiosis-related pea genes modulate fungal and plant gene expression during the arbuscule stage of mycorrhiza with Glomus intraradices.

The arbuscular mycorrhiza association results from a successful interaction between genomes of the plant and fungal symbiotic partners. In this study, we analyzed the effect of inactivation of late-stage symbiosis-related pea genes on symbiosis-associated fungal and plant molecular responses in order to gain insight into their role in the functional mycorrhizal association. The expression of a subset of ten fungal and eight plant genes, previously reported to be activated during mycorrhiza development, was compared in Glomus intraradices-inoculated wild-type and isogenic genotypes of pea mutated for the PsSym36, PsSym33, and PsSym40 genes where arbuscule formation is inhibited or fungal turnover modulated, respectively. Microdissection was used to corroborate arbuscule-related fungal gene expression. Molecular responses varied between pea genotypes and with fungal development. Most of the fungal genes were downregulated when arbuscule formation was defective, and several were upregulated with more rapid fungal development. Some of the plant genes were also affected by inactivation of the PsSym36, PsSym33, and PsSym40 loci, but in a more time-dependent way during root colonization by G. intraradices. Results indicate a role of the late-stage symbiosis-related pea genes not only in mycorrhiza development but also in the symbiotic functioning of arbuscule-containing cells.

Seasonal Migrations of Pantala flavescens (Odonata: Libellulidae) in Middle Asia and Understanding of the Migration Model in the Afro-Asian Region Using Stable Isotopes of Hydrogen.

In Middle Asia, the dragonfly Pantala flavescens makes regular seasonal migrations. In spring, sexually mature dragonflies (immigrants) arrive in this region for reproduction. Dragonflies of the aboriginal generation (residents) develop in about two months, and migrate south in autumn. Residents of Middle Asia have significantly lower δ2H values (-123.5 (SD 17.2)‱, n = 53) than immigrants (-64.4 (9.7)‱, n = 12), as well as aboriginal dragonfly species from Ethiopia (-47.9 (10.8)‱, n = 4) and the Sahel zone (-50.1 (15.5)‱, n = 11). Phenological data on P. flavescens in the Afro-Asian region and a comparison with published isotopic data on migratory insects from this region suggest that (i) the probable area of origin of P. flavescens immigrants is located in tropical parts of East Africa and/or the Arabian Peninsula and (ii) the autumn migration of Middle Asian residents to the south may also pass through the Indian Ocean. We assume that in the Afro-Asian region, there is an extensive migration circle of P. flavescens covering East Africa, Central Asia and the Indian subcontinent with a total length of more than 14,000 km.

Initiation of a legume nodule with an indeterminate meristem involves proliferating host cells that harbour infection threads.

A comparative analysis of nodule morphogenesis was carried out for three symbiotically defective pea (Pisum sativum) mutants that show abnormalities in nodule development.In the wild-type lines, resumption of cell proliferation in the pericycle and inner cortex results in the development of a nodule primordium, within which are found proliferating cells that harbour infection threads. However, this class of cell is not observed in the mutants RisFixA (sym41) and SGEFix­-2 (sym33) where nodule development is arrested at the point of formation of the apical nodule meristem. Itis proposed that the presence of proliferating cells harbouring infection threads is a prerequisite for normal formation of the nodule meristem.In mutant SGEFix­-1 (sym40), nodule development does not differ from that of wild-type plants in the early stages but is blocked at the stage after nodule meristem persistence. A scheme is proposed for the sequential functioning of pea symbiotic genes Sym33, Sym40 and Sym41 in the programme of nodule development.

New sources of Sym2A allele in the pea (Pisum sativum L.) carry the unique variant of candidate LysM-RLK gene LykX.

At the onset of legume-rhizobial symbiosis, the mutual recognition of partners occurs based on a complicated interaction between signal molecules and receptors. Bacterial signal molecules named Nod factors ("nodulation factors") are perceived by the plant LysM-containing receptor-like kinases (LysM-RLKs) that recognize details of its structure (i.e., unique substitutions), thus providing the conditions particular to symbiosis. In the garden pea (Pisum sativum L.), the allelic state of Sym2 gene has long been reported to regulate the symbiotic specificity: for infection to be successful, plants with the Sym2 A allele (for "Sym2 Afghan", as these genotypes originate mostly from Afghanistan) require an additional acetylation of the Nod factor which is irrelevant for genotypes with the Sym2 E allele (for "Sym2 European"). Despite being described about 90 years ago, Sym2 has not yet been cloned, though phenotypic analysis suggests it probably encodes a receptor for the Nod factor. Recently, we described a novel pea gene LykX (PsLykX) from the LysM-RLK gene family that demonstrates a perfect correlation between its allelic state and the symbiotic specificity of the Sym2 A-type. Here we report on a series of Middle-Eastern pea genotypes exhibiting the phenotype of narrow symbiotic specificity discovered in the VIR plant genetic resources gene bank (Saint-Petersburg, Russia). These genotypes are new sources of Sym2 A, as has been confirmed by an allelism test with Sym2 A pea cv. Afghanistan. Within these genotypes, LykX is present either in the allelic state characteristic for cv. Afghanistan, or in another, minor allelic state found in two genotypes from Tajikistan and Turkmenistan. Plants carrying the second allele demonstrate the same block of rhizobial infection as cv. Afghanistan when inoculated with an incompatible strain. Intriguingly, this "Tajik" allele of LykX differs from the "European" one by a single nucleotide polymorphism leading to an R75P change in the receptor part of the putative protein. Thus, our new data are in agreement with the hypothesis concerning the identity of LykX and the elusive Sym2 gene.

Application of multiphoton imaging and machine learning to lymphedema tissue analysis.

The results of in-vivo two-photon imaging of lymphedema tissue are presented. The study involved 36 image samples from II stage lymphedema patients and 42 image samples from healthy volunteers. The papillary layer of the skin with a penetration depth of about 100 µm was examined. Both the collagen network disorganization and increase of the collagen/elastin ratio in lymphedema tissue, characterizing the severity of fibrosis, was observed. Various methods of image characterization, including edge detectors, a histogram of oriented gradients method, and a predictive model for diagnosis using machine learning, were used. The classification by "ensemble learning" provided 96% accuracy in validating the data from the testing set.

Diagnosis of oral lichen planus from analysis of saliva samples using terahertz time-domain spectroscopy and chemometrics.

The ability to diagnose oral lichen planus (OLP) based on saliva analysis using THz time-domain spectroscopy and chemometrics is discussed. The study involved 30 patients (2 male and 28 female) with OLP. This group consisted of two subgroups with the erosive form of OLP (n = 15) and with the reticular and papular forms of OLP (n = 15). The control group consisted of six healthy volunteers (one male and five females) without inflammation in the mucous membrane in the oral cavity and without periodontitis. Principal component analysis was used to reveal informative features in the experimental data. The one-versus-one multiclass classifier using support vector machine binary classifiers was used. The two-stage classification approach using several absorption spectra scans for an individual saliva sample provided 100% accuracy of differential classification between OLP subgroups and control group.

Massive Analysis of cDNA Ends (MACE) for transcript-based marker design in pea (Pisum sativum L.).

Aimed at gene-based markers design, we generated and analyzed transcriptome sequencing datasets for six pea (Pisum sativum L.) genetic lines that have not previously been massively genotyped. Five cDNA libraries obtained from nodules or nodulated roots of genetic lines Finale, Frisson, Sparkle, Sprint-2 and NGB1238 were sequenced using a versatile 3'-RNA-seq protocol called MACE (Massive Analysis of cDNA Ends). MACE delivers a single next-generation sequence from the 3'-end of each individual cDNA molecule that precisely quantifies the respective transcripts. Since the contig generated from the 3'-end of the cDNA by assembling all sequences encompasses the highly polymorphic 3'-untranslated region (3'-UTR), MACE efficiently detects single nucleotide variants (SNVs). Mapping MACE reads to the reference nodule transcriptome assembly of the pea line SGE (Transcriptome Shotgun Assembly GDTM00000000.1) resulted in characterization of over 34,000 polymorphic sites in more than 9700 contigs. Several of these SNVs were located within recognition sequences of restriction endonucleases which allowed the design of co-dominant CAPS markers for the particular transcript. Cleaned reads of sequenced libraries are available from European Nucleotide Archive (http://www.ebi.ac.uk/) under accessions PRJEB18101, PRJEB18102, PRJEB18103, PRJEB18104, PRJEB17691.

Exhaled air analysis using wideband wave number tuning range infrared laser photoacoustic spectroscopy.

The infrared laser photoacoustic spectroscopy (LPAS) and the pattern-recognition-based approach for noninvasive express diagnostics of pulmonary diseases on the basis of absorption spectra analysis of the patient's exhaled air are presented. The study involved lung cancer patients ( N = 9 ), patients with chronic obstructive pulmonary disease ( N = 12 ), and a control group of healthy, nonsmoking volunteers ( N = 11 ). The analysis of the measured absorption spectra was based at first on reduction of the dimension of the feature space using principal component analysis; thereafter, the dichotomous classification was carried out using the support vector machine. The gas chromatography­mass spectrometry method (GC­MS) was used as the reference. The estimated mean value of the sensitivity of exhaled air sample analysis by the LPAS in dichotomous classification was not less than 90% and specificity was not less than 69%; the analogous results of analysis by GC­MS were 68% and 60%, respectively. Also, the approach to differential diagnostics based on the set of SVM classifiers usage is presented.

De Novo Assembly of the Pea (Pisum sativum L.) Nodule Transcriptome.

The large size and complexity of the garden pea (Pisum sativum L.) genome hamper its sequencing and the discovery of pea gene resources. Although transcriptome sequencing provides extensive information about expressed genes, some tissue-specific transcripts can only be identified from particular organs under appropriate conditions. In this study, we performed RNA sequencing of polyadenylated transcripts from young pea nodules and root tips on an Illumina GAIIx system, followed by de novo transcriptome assembly using the Trinity program. We obtained more than 58,000 and 37,000 contigs from "Nodules" and "Root Tips" assemblies, respectively. The quality of the assemblies was assessed by comparison with pea expressed sequence tags and transcriptome sequencing project data available from NCBI website. The "Nodules" assembly was compared with the "Root Tips" assembly and with pea transcriptome sequencing data from projects indicating tissue specificity. As a result, approximately 13,000 nodule-specific contigs were found and annotated by alignment to known plant protein-coding sequences and by Gene Ontology searching. Of these, 581 sequences were found to possess full CDSs and could thus be considered as novel nodule-specific transcripts of pea. The information about pea nodule-specific gene sequences can be applied for gene-based markers creation, polymorphism studies, and real-time PCR.

Coupling of CpCr(CO)3 and heterocyclic dithiadiazolyl radicals. synthetic, x-ray diffraction, dynamic NMR, EPR, CV, and DFT studies.

The reaction of the 1,2,3,5-dithiadiazolyls (4-R-C(6)H(4)CN(2)S(2))(2) (R = Me, 2a; Cl, 2b; OMe, 2c; and CF3, 2d) and (3-NC-5-tBu-C(6)H(3)CN(2)S(2))(2) (2e) with [CpCr(CO)(3)](2) (Cp = eta(5)-C(5)H(5)) (1) at ambient temperature respectively yielded the complexes CpCr(CO)(2)(eta(2)-S(2)N(2)CC(6)H(4)R) (R = 4-Me, 3a; 4-Cl, 3b; 4-OMe, 3c; and 4-CF(3), 3d) and CpCr(CO)(2)(eta(2)-S(2)N(2)CC(6)H(3)-3-(CN)-5-(tBu)) (3e) in 35-72% yields. The complexes 3c and 3d were also synthesized via a salt metathesis method from the reaction of NaCpCr(CO)(3) (1B) and the 1,2,3,5-dithiadiazolium chlorides 4-R-C(60H(4)CN(2)S(2)Cl (R = OMe, 8c; CF(3), 8d) with much lower yields of 6 and 20%, respectively. The complexes were characterized spectroscopically and also by single-crystal X-ray diffraction analysis. Cyclic voltammetry experiments were conducted on 3a-e, EPR spectra were obtained of one-electron-reduced forms of 3a-e, and variable temperature 1H NMR studies were carried out on complex 3d. Hybrid DFT calculations were performed on the model system [CpCr(CO)(2)S(2)N(2)CH] and comparisons are made with the reported CpCr(CO)(2)(pi-allyl) complexes.

A chemically induced new pea (Pisum sativum) mutant SGECdt with increased tolerance to, and accumulation of, cadmium.

BACKGROUND AND AIMS: To date, there are no crop mutants described in the literature that display both Cd accumulation and tolerance. In the present study a unique pea (Pisum sativum) mutant SGECd(t) with increased Cd tolerance and accumulation was isolated and characterized. METHODS: Ethylmethane sulfonate mutagenesis of the pea line SGE was used to obtain the mutant. Screening for Cd-tolerant seedlings in the M2 generation was performed using hydroponics in the presence of 6 microm CdCl2. Hybridological analysis was used to identify the inheritance of the mutant phenotype. Several physiological and biochemical characteristics of SGECd(t) were studied in hydroponic experiments in the presence of 3 microm CdCl2, and elemental analysis was conducted. KEY RESULTS: The mutant SGECd(t) was characterized as having a monogenic inheritance and a recessive phenotype. It showed increased Cd concentrations in roots and shoots but no obvious morphological defects, demonstrating its capability to cope well with increased Cd levels in its tissues. The enhanced Cd accumulation in the mutant was accompanied by maintenance of homeostasis of shoot Ca, Mg, Zn and Mn contents, and root Ca and Mg contents. Through the application of La(+3) and the exclusion of Ca from the nutrient solution, maintenance of nutrient homeostasis in Cd-stressed SGECd(t) was shown to contribute to the increased Cd tolerance. Control plants of the mutant (i.e. no Cd treatment) had elevated concentrations of glutathione (GSH) in the roots. Through measurements of chitinase and guaiacol-dependent peroxidase activities, as well as proline and non-protein thiol (NPT) levels, it was shown that there were lower levels of Cd stress both in roots and shoots of SGECd(t). Accumulation of phytochelatins [(PCcalculated) = (NPT)-(GSH)] could be excluded as a cause of the increased Cd tolerance in the mutant. CONCLUSIONS: The SGECd(t) mutant represents a novel and unique model to study adaptation of plants to toxic heavy metal concentrations.

On the motion of a heavy rigid body in an ideal fluid with circulation.

We consider Chaplygin's equations [Izd. Akad. Nauk SSSR 3, 3 (1933)] describing the planar motion of a rigid body in an unbounded volume of an ideal fluid while circulation around the body is not zero. Hamiltonian structures and new integrable cases are revealed; certain remarkable partial solutions are found and their stability is examined. The nonintegrability of the system describing the motion of a body in the field of gravity is proved and the chaotic behavior of the system is illustrated.