The Role of Medicago lupulina Interaction with Rhizophagus irregularis in the Determination of Root Metabolome at Early Stages of AM Symbiosis.

The nature of plant-fungi interaction at early stages of arbuscular mycorrhiza (AM) development is still a puzzling problem. To investigate the processes behind this interaction, we used the Medicago lupulina MlS-1 line that forms high-efficient AM symbiosis with Rhizophagus irregularis. AM fungus actively colonizes the root system of the host plant and contributes to the formation of effective AM as characterized by a high mycorrhizal growth response (MGR) in the host plant. The present study is aimed at distinguishing the alterations in the M. lupulina root metabolic profile as an indicative marker of effective symbiosis. We examined the root metabolome at the 14th and 24th day after sowing and inoculation (DAS) with low substrate phosphorus levels. A GS-MS analysis detected 316 metabolites. Results indicated that profiles of M. lupulina root metabolites differed from those in leaves previously detected. The roots contained fewer sugars and organic acids. Hence, compounds supporting the growth of mycorrhizal fungus (especially amino acids, specific lipids, and carbohydrates) accumulated, and their presence coincided with intensive development of AM structures. Mycorrhization determined the root metabolite profile to a greater extent than host plant development. The obtained data highlight the importance of active plant-fungi metabolic interaction at early stages of host plant development for the determination of symbiotic efficiency.

Mycorrhiza-Induced Alterations in Metabolome of Medicago lupulina Leaves during Symbiosis Development.

The present study is aimed at disclosing metabolic profile alterations in the leaves of the Medicago lupulina MlS-1 line that result from high-efficiency arbuscular mycorrhiza (AM) symbiosis formed with Rhizophagus irregularis under condition of a low phosphorus level in the substrate. A highly effective AM symbiosis was established in the period from the stooling to the shoot branching initiation stage (the efficiency in stem height exceeded 200%). Mycorrhization led to a more intensive accumulation of phosphates (glycerophosphoglycerol and inorganic phosphate) in M. lupulina leaves. Metabolic spectra were detected with GS-MS analysis. The application of complex mathematical analyses made it possible to identify the clustering of various groups of 320 metabolites and thus demonstrate the central importance of the carbohydrate and carboxylate-amino acid clusters. The results obtained indicate a delay in the metabolic development of mycorrhized plants. Thus, AM not only accelerates the transition between plant developmental stages but delays biochemical "maturation" mainly in the form of a lag of sugar accumulation in comparison with non-mycorrhized plants. Several methods of statistical modeling proved that, at least with respect to determining the metabolic status of host-plant leaves, stages of phenological development have priority over calendar age.

Metabolic Alterations in Pisum sativum Roots during Plant Growth and Arbuscular Mycorrhiza Development.

Intensive exchange of nutrients is a crucial part of the complex interaction between a host plant and fungi within arbuscular mycorrhizal (AM) symbiosis. For the first time, the present study demonstrates how inoculation with AMF Rhizophagus irregularis affects the pea (Pisum sativum L.) root metabolism at key stages of plant development. These correspond to days 21 (vegetation), 42 (flowering initiation), and 56 (fruiting-green pod). Metabolome profiling was carried out by means of a state-of-the-art GC-MS technique. The content shifts revealed include lipophilic compounds, sugars, carboxylates, and amino acids. The metabolic alterations were principally dependent on the stage of plant development but were also affected by the development of AM fungi, a fact which highlights interaction between symbiotic partners. The comparison of the present data with the results of leaf metabolome profiling earlier obtained did not reveal common signatures of metabolic response to mycorrhization in leaves and roots. We supposed that the feedback for the development and symbiotic interaction on the part of the supraorganismic system (root + AM fungi) was the cause of the difference between the metabolic profile shift in leaf and root cells that our examination revealed. New investigations are required to expand our knowledge of metabolome plasticity of the whole organism and/or system of organisms, and such results might be put to use for the intensification of sustainable agriculture.

EVIDENCE-BASED MEDICINE AS PATIENT'S PROTECTION MEASURE IN JUDICIAL PRACTICE.

OBJECTIVE: The aim: To identify the role of evidence-based medicine, its principles and approaches to patients' rights protection and the provision of medical service optimization, the skills of medical staff improvement, increasing the objectivity of court decisions in cases of non-providing quality care or death. PATIENTS AND METHODS: Materials and methods: The authors used the decisions by the European Court of Human Rights (ECtHR) on the statements of victims of unprofessional actions by doctors, international and domestic regulations on patients' rights, statistics on the results of criminal proceedings on violations of patients' rights over the past 5 years, case law of criminal and civil jurisdiction in this category of cases, the results of surveys of prosecutors, as well as the results of research by scientists in the field of medical law and criminalistics. The research is carried out on the basis of a harmonious combination of philosophical approaches, general scientific and special methods of scientific knowledge, the complex of which is chosen taking into account the goals and objectives, object and subject of research. CONCLUSION: Conclusions: In order to implement the patients' rights by legal means in accordance with the Convention for the Protection of Human Rights and Fundamental Freedoms, the European Charter of Patients' Rights and other international regulations in the field of medicine, the authors argue the need for greater implementation of evidence-based medicine into the practice of medical institutions as a means of improving the level of medical care and an obvious source of relevant information for litigation to protect the rights of patients and doctors. For patients' rights protection in the diagnosis and treatment of diseases, it is proposed to regulate a set of such measures: 1) to include in the educational programs of pharmacy and medicine such disciplines as «Evidence-Based Medicine¼ and «Rights of the patient and medical worker's rights¼; 2) to oblige the doctor to explain to the patient or to his/her representative the differences of treatment protocols, to provide information to patients about official sources, which contain information on unified and updated treatment protocols and diagnosis of certain diseases, to obtain informed consent by the patient (his/her representative) on certain medical guideline; 3) to recognize medical guidelines as sources of law in criminal and civil proceedings as a kind of benchmarks for clarifying and assessing the facts of non-performance or improper performance of professional duties by a medical or pharmaceutical worker, violation of patients' rights, as well as means for doctors' legal protection and etc.

Metabolic alterations in pea leaves during arbuscular mycorrhiza development.

Arbuscular mycorrhiza (AM) is known to be a mutually beneficial plant-fungal symbiosis; however, the effect of mycorrhization is heavily dependent on multiple biotic and abiotic factors. Therefore, for the proper employment of such plant-fungal symbiotic systems in agriculture, a detailed understanding of the molecular basis of the plant developmental response to mycorrhization is needed. The aim of this work was to uncover the physiological and metabolic alterations in pea (Pisum sativum L.) leaves associated with mycorrhization at key plant developmental stages. Plants of pea cv. Finale were grown in constant environmental conditions under phosphate deficiency. The plants were analyzed at six distinct time points, which corresponded to certain developmental stages of the pea: I: 7 days post inoculation (DPI) when the second leaf is fully unfolded with one pair of leaflets and a simple tendril; II: 21 DPI at first leaf with two pairs of leaflets and a complex tendril; III: 32 DPI when the floral bud is enclosed; IV: 42 DPI at the first open flower; V: 56 DPI when the pod is filled with green seeds; and VI: 90-110 DPI at the dry harvest stage. Inoculation with Rhizophagus irregularis had no effect on the fresh or dry shoot weight, the leaf photochemical activity, accumulation of chlorophyll a, b or carotenoids. However, at stage III (corresponding to the most active phase of mycorrhiza development), the number of internodes between cotyledons and the youngest completely developed leaf was lower in the inoculated plants than in those without inoculation. Moreover, inoculation extended the vegetation period of the host plants, and resulted in increase of the average dry weight per seed at stage VI. The leaf metabolome, as analyzed with GC-MS, included about three hundred distinct metabolites and showed a strong correlation with plant age, and, to a lesser extent, was influenced by mycorrhization. Metabolic shifts influenced the levels of sugars, amino acids and other intermediates of nitrogen and phosphorus metabolism. The use of unsupervised dimension reduction methods showed that (i) at stage II, the metabolite spectra of inoculated plants were similar to those of the control, and (ii) at stages IV and V, the leaf metabolic profiles of inoculated plants shifted towards the profiles of the control plants at earlier developmental stages. At stage IV the inoculated plants exhibited a higher level of metabolism of nitrogen, organic acids, and lipophilic compounds in comparison to control plants. Thus, mycorrhization led to the retardation of plant development, which was also associated with higher seed biomass accumulation in plants with an extended vegetation period. The symbiotic crosstalk between host plant and AM fungi leads to alterations in several biochemical pathways the details of which need to be elucidated in further studies.

Independent forensic medical examination as a mean of proving the facts of a torture usage.

OBJECTIVE: Introduction: In most countries detainees are often subjected to physical and mental abuse by law enforcement officials, but very rarely victims of torture can prove the guilt of these offenders due to the poor quality of findings of state forensic medical and forensic psychiatric experts and due to inconsistency of their findings with international guidelines (Istanbul Protocol). The aim: To determine the role of forensic medical examination in the investigation of torture crimes, to provide arguments for necessity to security of the victim's right to collect evidence independently, including through using of special knowledge of independent forensic medical and forensic psychiatric experts in criminal proceedings. PATIENTS AND METHODS: Materials and methods: The authors used the European Court of Human Rights (ECHR) Decisions on the complaints of the victims' torture, international and Ukrainian human rights legal acts, the results of numerous torture investigations conducted by medical and criminalistics scientists. The research is carried out on the basis of a harmonious combination of philosophical approaches, general and special scientific methods. CONCLUSION: Conclusions: In order to exercise the rights of victims of torture in accordance with the Convention on Human Rights and Fundamental Freedoms, the Convention against Torture and Other Cruel, Inhuman or Degrading Treatment or Punishment, and according to other international and Ukrainian legal acts on human rights the arguments given for the need to enable the victim to engage independent forensic medical and forensic psychiatric experts to provide their conclusions in torture-related criminal proceedings.