A Model of Acute Peritonitis.

The biological models used in the study of generalized peritonitis can be subdivided into 5 groups (introduction of foreign bodies, cultures of microorganisms, suspensions of feces, chemicals, and mechanical damage to the gastrointestinal tract) or into 4 groups (introduction of foreign bodies, chemicals, bacterial contamination of the abdominal cavity, and combined methods). After analysis of published reports, the most justified classification of methods of peritonitis modelling is based on the type of peritonitis-inducing agent and the administration route and on the nature of peritonitis developing in the abdominal cavity. The choice of the model maximally close reproducing clinical conditions of peritonitis should be based on the specific objectives of the study, focusing on the etiology, pathogenesis, and severity of the disease course, planned measures aimed at eliminating the process, and other factors.

Simulation of Mechanical and Thermal Wounds of Soft Tissues.

Millions of accidental and surgical injuries of soft tissues are registered annually around the world [5]. Untimely and insufficiently effective treatment of wounds in 50-70% leads to the development of purulent-septic infection and the development of septic conditions and fatal outcomes [1-4], which necessitates thorough study of inflammatory and regenerative processes occurring in the injured soft tissues. Various models of mechanical and thermal damage to soft tissues are proposed for studying the inflammatory and reparative processes, for assessing the therapeutic effects and developing new approaches to wound treatment. However, the developed models do not fully meet the requirements of researchers and are not always simple and uniformly reproducible, close to the course of the pathology in humans, and highly reliable. When choosing the model of mechanical and thermal wounds, the experience of other researchers should be taken into consideration due to the need of actualization and improvement of existing models.

Generation of an induced pluripotent stem cell line ICGi030-A from a Wilson's disease patient carrying a frameshift mutation p.Lys1013fs and missense mutation p.H1069Q in the ATP7B gene.

Wilson's disease is a rare autosomal recessive disorder of copper metabolism. The copper accumulation in the viscera appears due to the functional impairment of copper-transporting ATPase, which is encoded by the ATP7B gene. In this study, PBMCs of a patient with two ATP7B mutations were reprogrammed. The first mutation is a missense mutation p.H1069Q, which is the most frequent mutation in the human population. At the same time, the second one is a frameshift mutation p.Lys1013fs. The generated iPSC line had a normal karyotype, maintained the original genotype, expressed pluripotency markers, and demonstrated the ability to differentiate into derivatives of the three germ layers.

Generation of induced pluripotent stem cell lines ICGi021-A and ICGi022-A from peripheral blood mononuclear cells of two healthy individuals from Siberian population.

ICGi021-A and ICGi022-A iPSC lines were obtained by reprogramming PBMCs of two healthy women of the Siberian population using episomal non-integrating vectors expressing Yamanaka factors. iPSC lines expressed pluripotency markers, had a normal karyotype and demonstrated the ability to differentiate into derivatives of the three germ layers. Clinical exome sequencing data of the original biosamples of the donors are available in the NCBI SRA database. The generated cell lines are useful as "healthy" control in biomedical studies.

Generation of two induced pluripotent stem cell lines from peripheral blood mononuclear cells of a patient with Wilson's disease.

Wilson's disease is an inherited disorder associated with copper accumulation in the liver, brain and other vital organs. Wilson's disease is caused by mutations in the ATP7B gene. Over 300 mutations of ATP7B have been described. Despite the disease is autosomal recessive, the patient whose PBMCs were reprogrammed in the study harbours heterozygous mutation c.3207C > A (p.H1069Q). Detailed analysis of the ATP7B complete gene sequencing data has not revealed other known disease associated mutation. The generated iPSC lines maintained the original genotype, expressed pluripotency markers, had normal karyotype and demonstrated the ability to differentiate into derivatives of the three germ layers.

Generation of induced pluripotent stem cell line ICGi018-A from peripheral blood mononuclear cells of a patient with Huntington's disease.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by CAG repeat expansion in the HTT gene. HD patient-specific induced pluripotent stem cells (iPSCs) represent an excellent model for the disease study. We generated iPSC line from blood mononuclear cells of HD patient with 38 CAG repeats in the HTT exon 1 using integration free episomal plasmids expressing Yamanaka factors. The iPSC line retained the disease causing mutation and expressed pluripotency markers. It also displayed a normal karyotype and the ability to differentiate into derivatives of three germ layers.

Generation of two iPSC lines, (ICGi015-A and ICGi015-B), by reprogramming peripheral blood mononuclear cells from a patient with Parkinson's disease.

Studying Parkinson's disease (PD), one of the most common neurodegenerative disorders worldwide, requires different model systems, including patient-specific induced pluripotent stem cell lines. With the help of non-integrating episomal vectors the iPSC lines ICGi015-A and ICGi015-B were generated from blood mononuclear cells of PD patient, carrying three SNPs, associated with PD development. The obtained iPSC lines express pluripotency markers and demonstrate the ability to in vitro differentiate into the three germ layers. These cell lines may be useful for studying molecular mechanisms of PD and for drug screening.

Generation of two iPSC lines (ICGi008-A and ICGi008-B) from skin fibroblasts of a patient with early-onset Alzheimer's disease caused by London familial APP mutation (V717I).

The induced pluripotent stem cell (iPSC) lines ICGi008-A and ICGi008-B were generated from dermal fibroblasts using episomal vectors expressing pluripotency factors. Dermal fibroblasts were obtained from a 55 year old male Сaucasian familial Alzheimer's disease (AD) patient carrying heterozygous V717I mutation in the APP gene. The generated iPSC lines maintained the original APP genotype, expressed pluripotency markers, exhibited a normal karyotype and retained the ability to differentiate into cell types of the three germ layers. The iPSC lines will be useful for the study of the AD molecular and cellular mechanisms and drug screening.

Generation of two spinal muscular atrophy (SMA) type I patient-derived induced pluripotent stem cell (iPSC) lines and two SMA type II patient-derived iPSC lines.

Spinal muscular atrophy (SMA) is a neuromuscular disease caused by deletion or mutation in SMN1 gene. SMA human induced pluripotent stem cells (iPSCs) represent a useful and valid model for the study of the disorder, as they provide in vitro the target cells. We generated iPSCs from a SMA type I patient and SMA type II patient by using non-integrating episomal plasmid vectors. The resulting iPSCs are episomal-free, express pluripotency markers, display a normal karyotype, retain the mutation (homozygous deletion of SMN1) and are able to differentiate into the three germ layers.

Generation of induced pluripotent stem cell line, ICGi007-A, by reprogramming peripheral blood mononuclear cells from a patient with Huntington's disease.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by mutation in the HTT gene encoding HTT protein. The mutant protein leads to the neuronal death through dysregulation of multiple cellular processes. HD human induced pluripotent stem cells (iPSCs) represent a useful and valid model for the disease study. iPSC line from HD patient with 47 CAG repeats in HTT was generated from blood mononuclear cells by non-integrating episomal vectors. The iPSC line retained the mutation, expressed pluripotency markers, had a normal karyotype and displayed in vitro differentiation to the three germ layers. Resource table.

[Surgical treatment patients with drug-resistant unilateral MRI-positive temporal forms of epilepsy]. / Khirurgicheskoe lechenie patsientov s farmakorezistentnymi MR-pozitivnymi simptomaticheskimi odnostoronnimi visochnymi formami épilepsii.

AIM: To evaluate the results of surgical treatment of patients with unilateral MRI-positive temporal forms of drug-resistant epilepsy. MATERIAL AND METHODS: A prospective analysis of 50 patients with unilateral MRI-positive temporal forms of drug-resistant epilepsy, who had undergone resective surgery in the Scientific Research Institute of Emergency Medicine of N.V. Sklifosovsky (Moscow) and in University Clinic of Moscow State University of Medicine and Dentistry between 01.01.14 and 12.12.17, has been performed. MRI-positive temporal symptomatic epilepsy was identified in 79 (55%) patients. MRI results showed unilateral temporal epileptogenic lesions in 50 (67%) patients. These patients were assigned to medial frontal lobectomy. RESULTS: 82% patients become 'seizure free' 12 and 24 months after surgical treatment. In other cases, seizures become less frequent and severe. CONCLUSION: The results confirm the efficacy and safety of surgical treatment of drug-resistant MRI-positive temporal forms of epilepsy.

[Surgical prevention of ischaemic strokes by means of an extraintracranial microanastomosis (EICMA) in the carotid basin]. / Khirurgicheskaya profilaktika ishemicheskikh insul'tov s ispol'zovaniem ekstra-intrakranial'nogo mikroanastomoza (EIKMA) v karotidnom basseine.

BACKGROUND: Not less than 50% of all ischaemic strokes appear to occur resulting from pathology of extracranial arteries. Occlusions and stenoses are more commonly encountered in carotid arteries, with the incidence of occlusion of the internal carotid artery (ICA) ranging from 5 to 10% within the structure of all lesions of brachiocephalic arteries (BCA). AIM: The study was aimed at assessing the results of a surgical procedure of extra-intracranial microanastomosis (EICMA) performed in patients presenting with occlusive and stenotic lesions of BCA at the Neurosurgical Department. PATIENTS AND METHODS: During the period between January 1st 2009 to September 30th 2015, specialists of the Neurosurgery Department of the Research Institute of Emergency Medical Care named after N.V. Sklifosovsky treated a total of 1,101 patients presenting with atherosclerotic lesions of BCA, with a total of 1,038 surgical interventions performed. Of these, there were 837 operations of carotid endarterectomy and 180 operations of EICMA in the carotid basin. The indications for performing EICMA in patients from the group with occlusive-stenosing lesions of the major arteries of the head and neck were as follows: the presence of occlusion of the internal carotid artery or M1 segment of the middle cerebral artery, verified by the findings of angiographic examinations; previously endured ischaemic-type cerebral circulation impairment in the basin on the side of occlusion; a decrease in the perfusion reserve of the brain on the side of occlusion by the findings of positron emission tomography (PET), single-photon emission computed tomography (SPECT) or CT-perfusion with loading tests. RESULTS: After performing EICMA, the neurological status in patients averagely improved by 1.2 points according to the NIH Stroke Scale; by 0.5 point according to the Rankin scale, and by 3.5 points according to the Rivermead Mobility Index (Table 1). The dynamics of functional outcomes of EICMA turned out to depend on the terms of the operation (R=2143, p<0.05) and the degree of neurological deficit prior to surgery (R=3422, p<0.05): the deeper the neurological deficiency was and the sooner after acute cerebral ischaemia (ACI) the operation was performed, the more significant restoration of the functions was at the moment of discharge from hospital. Major complications of EICMA in patients operated on at various terms of ischaemic stroke were as follows: recurrent cerebral circulatory impairments - in 6 (3.3%) patients, subdural and intracranial haemorrhage - in 2 (1.1%), temporal deepening of the neurological deficit on the background of reperfusion, confirmed by the findings of SPECT - in 2 (1.1%), pneumonia with a lethal outcome - in 1 (0.6%), fatal ischaemic stroke of the opposite hemisphere - in 1 (0.6%) subject. The lethality rate amounted to 1.1% (2 cases). The risk for the development of complications was higher in elderly and aged patients (75 years and over), in patients with pronounced neurological deficit and significant somatic disorders. CONCLUSION: The indications for surgical prevention of recurrent ischaemic stroke by means of EICMA should be based on the following factors: the presence of confirmed occlusion of the ICA, a history of endured acute cerebral ischaemia, and decreased cerebrovascular reserve. Performing a surgical intervention is not appropriate for patients presenting with pronounced neurological deficit and significant somatic pathology. Careful selection of patients, the use of modern methods of neuroimaging, and intraoperative control make it possible to achieve favourable functional outcomes and to decrease the postoperative complications rate.

[The use of combined revascularization surgery for successful elimination of a middle cerebral artery aneurysm].

OBJECTIVE: The study objective is to present a clinical case of successful surgical treatment of a complex middle cerebral artery (MCA) aneurysm using various types of bypasses. MATERIAL AND METHODS: A 59-year-old female patient presented with nontraumatic intracranial hemorrhage caused by rupture of a complex right MCA aneurysm. The anatomical features of the MCA aneurysm were identified using computed tomography (CT) in angiographic and 3D modes. The surgical intervention included aneurysmectomy and an end-to-end reanastomosis between the M1 and M2 segments of the MCA followed by an extra-intracranial microvascular anastomosis (EICMA) between the frontal branch of the right superficial temporal artery (STA) and the cortical branch of the right MCA located on the frontal lobe surface. RESULTS: The intraoperative blood flow via an intra-intracranial bypass (IC-IC bypass) was 30 mL/min, and the linear velocity of blood flow (LVBF) was 50 cm/s; the blood flow and LVBF via the STA-MCA bypass were 7-8 mL/min and 15 cm/s, respectively. CT angiography performed on the 1st postoperative day revealed the patency of the IC-IC and STA-MCA bypasses. The patient was discharged in satisfactory condition (Glasgow Outcome Scale -V) 1 month after surgery. CONCLUSION: Revascularization surgery is the sought-after surgical technique for complex intracranial aneurysms that enables efficient exclusion of the aneurysm from blood flow and prevention of ischemic brain injuries in the carrying artery territory.

[The use of extra-intracranial microanastomosis in the treatment of cerebral ischemia in patients with nontraumatic subarachnoid hemorrhage]. / Primenenie ekstra-intrakranial'nogo mikroanastomoza v lechenii ishemii golovnogo mozga u bol'nykh s netravmaticheskim subarakhnoidal'nym krovoizliyaniem.

OBJECTIVE: to evaluate the use of extra-intracranial microanastomosis (EICMA) in the treatment of brain ischemia in patients with nontraumatic subarachnoid hemorrhage (SAH). MATERIAL AND METHODS: In the period from 01.01.14 to 01.07.15, there were 229 surgeries for ruptured intracranial aneurysms performed in the urgent surgery unit. Nine patients with marked and widespread angiospasm, subcompensated and decompensated cerebral ischemia underwent the simultaneous clipping of ruptured intracranial aneurysms and EICMA. The age of patients varied from 32 to 52 years (mean 36 years). The severity of patient's state was assessed as III-IV grades on the Hunt and Hess scale before operation. The surgery was performed 1-2 days after admission to the hospital, 1-8 days after the development of SAH. RESULTS AND CONCLUSION: Excellent and good outcome was recorded in 4 patients, severe disability in 3 patients, fatal outcome in 2 patients. The fatal outcome was due to decompensated cerebral ischemia and progressive angiospasm with the high linear blood flow rate and the following reduction in perfusion in the affected hemisphere. The simultaneous clipping of ruptured intracranial aneurysms and EICMA in the acute stage of SAH of patients with subcompensated cerebral ischemia allow to improve treatment RESULTS: This technique is most applicable for patients with proximal angiospasm of M1- and M2-segments of the middle cerebral artery in the first 24 h of the development of a focal neurological deficit supported by the reduction in perfusion in the corresponding vascular area.

[Combined subchronic toxicity of nickel and manganese oxides nanoparticles, and its decrease due to bioprotectors complex].

Stable suspensions of NiO and/or Mn304 nanoparticles with average diameter 16,7?8,2 nm and 18,4?5,4 nm respectively, obtained via laser ablation of the metals with 99,99% purification in deionized water, were injected intraperitoneally into rats in dose of 0,5 mg or 0,25 mg three times per week up to 18 times separately or in various dose combinations. A group of rats received combined injections of nanoparticles in the highest dose or merely water with oral <> containing pectin, vitamins A, C and E, glutamate, glycine, N-acetylcysteine, selenium, iodine and polyunsaturated fatty acids of omega-3 class. Intoxication development was assessed through multiple functional parameters and histologic changes in liver, spleen, kidneys and brain. Nickel and manganese accumulation in these organs was measured-via various methods. Both types of metallic oxide nanoparticles appeared to be hazardous for body, but Mn304 caused more harm according to major nonspecific toxicity manifestations. Moreover, they caused more intense injury to caudate nucleus and hippocamp neurons - that can be considered as an experimental model of manganese parkinsonism. Mathematic analysis based on response pattern revealed ambiguity of the combined toxicity type, depending on the effects assessed and on its level. Due to the bioprotector complex, organic and systemic toxicity and genotoxicity of Mn304 and NiO nanoparticles combined were diminished.

[Assessment of cerebral hemodynamics using computed tomography perfusion in patients with intracranial aneurysms].

OBJECTIVE: To determine the impact of aneurysm of intracranial arteries on cerebral collateral circulation and to assess the relationship between these changes and the site of the aneurysm. MATERIAL AND METHODS: CT perfusion and CT angiography were performed in 41 patients with 47 unruptured intracranial aneurysms of different sites and sizes. RESULTS: Impaired perfusion was not found in the majority of the patients with intracranial aneurysms less than 15 mm in diameter. Hypoperfusion in the respective temporal and occipital lobes was noted in 70% of patients with aneurysms greater than 16-25 mm. Generalized hypoperfusion was diagnosed in 100% of patients with aneurysms more than 25 mm, with the partially thrombosed aneurysms accompanied by the severer, irreversible deficit immediately around the aneurysmal sac, which was induced by the effect of volumetric exposure while in the presence of unthrombosed aneurysm, hypoperfusion occasionally involved the cortex of the entire hemisphere. In this investigation, the extent and depth of hypoperfusion depended on the thrombosis of the lumen and the size of an aneurysm rather than the site of the latter. CONCLUSION: CT perfusion may be recommended as a standard examination of patients with detected intracranial aneurysm that is 16 mm or more in size for the timely diagnosis of cortical microcirculatory disorders and collateral circulatory insufficiency, which can affect the choice of a treatment option for the patient.

Induced Pluripotent Stem Cells of Microtus levis x Microtus arvalis Vole Hybrids: Conditions Necessary for Their Generation and Self-Renewal.

Every year, the list of mammalian species for which cultures of pluripotent stem cells (PSCs) are generated increases. PSCs are a unique tool for extending the limits of experimental studies and modeling different biological processes. In this work, induced pluripotent stem cells (iPSCs) from the hybrids of common voles Microtus levis and Microtus arvalis, which are used as model objects to study genome organization on the molecular-genetic level and the mechanisms of X-chromosome inactivation, have been generated. Vole iPSCs were isolated and cultured in a medium containing cytokine LIF, basic fibroblast growth factor (bFGF), ascorbic acid, and fetal bovine serum. Undifferentiated state of vole iPSCs is maintained by activation of their endogenous pluripotency genes - Nanog, Oct4, Sox2, Sall4, and Esrrb. The cells were able to maintain undifferentiated state for at least 28 passages without change in their morphology and give rise to three germ layers (ectoderm, mesoderm and endoderm) upon differentiation.

[CT perfusion in patients after EICMA in the postoperative period].

OBJECTIVE: To evaluate the influence of the EICMA on the circulation of the brain in patients with unilateral occlusion of the internal carotid artery (ICA) in the late postoperative period using CT perfusion. MATERIAL AND METHODS: Ten patients with unilateral internal carotid artery occlusion and stenosis of the opposite ICA 50-60%, with a history of a single ischemic stroke were examined. All patients underwent pre-and postoperative CT angiography of the brachiocephalic and intracranial arteries and CT perfusion. We also compared the results of neurological examinations preoperatively and during the year after the intervention. RESULTS: The neurological data during the first year after surgery demonstrated an improvement of neurological status and quality of life in all patients. Preoperative CT perfusion showed the patchy decrease in the cerebral blood flow (CBF) to 18 ml/100 g/min (average of 44-56 ml/100 g/min) and increase in the mean transit time (MTT) to 7.2 s (normally less 6c) in all cases on the side of occlusion. The most susceptible to chronic ischemic changes was the frontal region, temporal and parietooccipital regions were affected to a lesser extent. Due to stenosis of the opposite ICA, minimum CBF of the cortex in the opposite hemisphere was 24 ml/100 g/min and MTT was increased to 5.6 s. Six months after the applying of EICMA, the significant improvement of CT perfusion was noted on the side of the anastomosis in all patients: an increase in CBF (at least 44 ml/100 g/min) and MTT reduction (up to 6.1s in the frontal region), as well as the "synchronization" of CBF and CBV in similar areas of the cerebral cortex of the right and left hemisphere. CONCLUSION: CT perfusion in the late postoperative period after applying EICMA in patients with unilateral ICA occlusion demonstrates not only changes of the cerebral perfusion on the side of the occlusion, but also the increased collateral blood flow of the cortex in both hemispheres, which significantly improves brain blood flow generally within 6-12 months after surgery.

[Certain toxicodynamic and toxicokinetic features of combined subchronic intoxication with hexavalent chromium and nickel].

Repeated intraperitoneal injections of nickel and chromium (VI) into rats appeared to demonstrate that the combined subchronic toxicity can be additive or vary (mostly to subadditivity) in accordance with effect on which they are evaluated. With moderate general toxic effects, the studied combination has marked genotoxicity with additive effect. The studies demonstrated reciprocal influence of nickel and chromium on accumulation of the second metal in some organs (especially, in spleen), but not on its renal excretion.

Characteristics of induced human pluripotent stem cells using DNA microarray technology.

We performed transcriptome analysis of some human induced pluripotent stem cells, embryonic stem cells, and human somatic cells using DNA microarrays. PluriTest bioinformatic system was used for evaluation of cell pluripotency. Changes in the genome structure and status of X-chromosome gene expression was analyzed using microarray technology.