The genes encoding of DNA ligases from the thermophilic archaeon Pyrococcus abyssi (PabDNA ligase) and Methanobacterium thermoautotrophicum (MthDNA ligase) were cloned and expressed in Escherichia coli. The activity of purified enzymes was studied by ligation of two oligonucleotides, one of which had preformed hairpin structure. In the used system the maximal output of reaction products for both DNA ligases was observed near 70 degrees C that is explained by substrate thermostability. At stoichiometric ratio of enzymes and substrate the output of a product reaches of plateau at 70-75% of theoretical ones. Investigated DNA ligases showed different thermostability. The half-time life of PabDNA ligase was about 60 min at 90 degrees C. MthDNA ligase was completely inactivated at this temperature during 10 min. Recombinant DNA ligases from P. abyssi and M. thermoautotrophicum possessed high stability during a storage at 4 degrees C.

Introduction. Critical limb ischemia (CLI), which arises from atherosclerotic lesions of the arteries is quite often presently. This problem is actively discussed all over the world. In addition to the disability of the patients due to limb loss, the death rate from this disease is comparable to mortality from malignant tumors. Objective. Evaluate immediate and long-term results and optimize treatment tactics for patients with critical limb ischemia in multilevel lesions of lower limb arteries. Materials and methods. The examination and treatment of patients with CLI was carried out in the Research Institute of Surgery and Urgent Medicine of the First State Medical University of Saint-Petersburg named by I. P. Pavlov, as well as on the basis of the City Clinical Hospital «City Hospital No. 14» and St. Petersburg Research Institute of Emergency Care named by I. I. Dzhanelidze in the period from 2005 to 2016. The study is conducted in patients with atherosclerotic lesion of the main arteries of the lower extremities, which manifests itself as pain at rest and/or tissue loss (stage 3-4 of chronic arterial insufficiency of the limb according to Fontein-Pokrovsky classification). Results. Clinical success of all kinds of interventions was achieved in 166 (88.8 %) patients. However, the negative results were greater in the group where only angioplasty of the lower limb arteries was performed. Long-term results could be traced in 108 (57.8 %) patients for 6 months - 5 years. If we consider all groups as a whole, then the aggravation of ischemia occurred in endovascular intervention group in 62 % of patients, in the group of bypass operations - in 19.61 %, and in the group of hybrid interventions - in 17.65 %. Conclusion. Hybrid methods of treatment are the method of choice for patients with stage 4 ischemia of the lower limb with multilevel lesions of lower extremity arteries. Open surgery on the arteries of the lower extremities remains a very effective method of treating patients with critical ischemia. In turn, the use of endovascular methods is advisable in patients with severe concomitant pathology.

Relevance. The relevance of the study is determined by the fact that hopes are placed in the cell therapy for patients with critical limb-threatening (CLI) ischemia as a method of the restoration of blood circulation in the affected limb in patients who cannot undergo surgical or endovascular intervention. Aim. To evaluate the efficiency of allogeneic MSCs for the treatment of critical lower limb ischemia (randomized placebo-controlled study).Materials and methods. The study included 34 patients with critical lower limb ischemia (grade 4 according to Pokrovsky). There were 18 patients in the MSC group, and 16 patients in the placebo group). The groups were comparable concerning age, disease duration, and comorbidities. Allogeneic MSCs (phenotype CD73+, CD90+, CD105+, CD45–, CD34–, CD14–) were injected into the posterior calf muscles. Clinical outcome, ankle pressure, transcutaneous oxygen tension (tcpO2), and pain-free walking distance (PFWD) were evaluated. The patients were followed-up for 12–36 months. According to the clinical outcome in each group, the patients were divided into subgroups with «effect (+)» or «effect (–)». In 2 patients, there was an «uncertain clinical outcome». When analyzing the results, these patients were assigned to one or another subgroup.Results. In the MSC and placebo groups, the clinical outcome assessed as «effect (+)» or «effect (–)» did not differ (OR 1.5; 95 % CI 0.34–6.7). With different variants of group formation and with the assignment of patients with an «uncertain clinical outcome» to a one or another subgroup, the final results neither differed. According to instrumental research methods (PFWD, tcpO2, ankle pressure, angiography), there were no differences in the MSC and placebo groups. Conclusion. With different variants of analysis and group formation, no convincing evidence that allogeneic MSCs can be effective for the treatment of critical lower limb ischemia have been obtained.

the latest primary care consultation and CV assessment are shown in Table 1. Whilst 2175 (66.7%) underwent a primary care consultation within 7-30 days of index, only 416 (12.8%) had a CV-assessment during the same period. Furthermore, 2073 (63.6%) patients had no CV-assessment within the 3 months prior to their amputation. Of this group, 1230 (59.3%) still underwent a primary care consultation for another reason in the 7-30 days prior to their amputation. Conclusion: 'Missed opportunities' for timely diagnosis of CLTI within general practice potentially exist. Further investigation is required to greater understand the reasoning for these opportunities being missed and to develop strategies to that would help prevent amputation.

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