Ex Vivo Production of IL-1ß and IL-10 by Activated Blood Cells of Wistar Rats with Different Resistance to Hypoxia after Systemic Inflammatory Response Syndrome.

We studied spontaneous and ex vivo activated cytokine production by blood cells of male Wistar rats with different resistance to hypoxia against the background of an LPS-induced systemic inflammatory response. In rats with low (LR) and high resistance (HR) to hypoxia, the number of leukocytes, granulocytes, and peripheral blood lymphocytes was determined, the levels of spontaneous and stimulated production of IL-1ß and IL-10 and their ratio were assessed ex vivo. Against the background of a systemic inflammatory response, only HR animals showed a decrease in spontaneous and stimulated production of IL-1ß and spontaneous production of IL-10. The IL-1ß/IL-10 ratio decreased only in LR rats during the development of a systemic inflammatory response, while in HR animals, no changes in this indicator were observed. The obtained data suggest a high proinflammatory potential of blood cells in LR rats, which apparently determines the development of a more severe course of the systemic inflammatory response.

[Physical therapy - the main focus of modern ophthalmic rehabilitation]. / Fizioterapiya ­ osnovnoe napravlenie sovremennoi oftal'moreabilitatsii.

Ophthalmic rehabilitation consists of improving or maintaining the functions of the visual analyzer within the possibilities of the "therapeutic window". Ophthalmic rehabilitation involves physiotherapeutic methods, as well as additional ones that improve the condition of the body and indirectly affect the organ of vision. This article presents schematic algorithms of physiotherapeutic ophthalmic stimulation in neurodystrophic diseases of the visual organ and the main results of their multifactorial objective and subjective analysis. It was shown that even with the consistent visual resolution, treatment courses can help get positive changes at the level of nerve structures lasting for three to six months. This allows us to recommend physiotherapeutic ophthalmic stimulation for maintaining the therapeutic effects obtained after the main medical or surgical treatment.

[Modern capabilities of ophthalmic tonometry in pathologically altered corneas]. / Sovremennye vozmozhnosti oftal'motonometrii pri patologicheskikh izmeneniyakh rogovitsy.

Changes in intraocular pressure have great influence on the course of various processes in ocular tissues. Presently, the scientific discussion has returned to the theoretical issues of tonometry and the creation of models that would allow assessing the influence of various ocular parameters on IOP values. Some of the modern tonometers (Icare, Tono-Pen) are convenient and effective when measuring IOP in eyes with damaged, uneven or edematous cornea. It is necessary to analyze the results of ophthalmic tonometry by modern methods in accordance with the data of direct intracameral manometry in pathological conditions of the cornea and after keratoplasty.

[Comparative evaluation of the results of one- and two-piece mushroom keratoplasty]. / Sravnitel'naya otsenka rezul'tatov mono- i dvukhkomponentnoi modifikatsii gribovidnoi keratoplastiki.

PURPOSE: To compare and evaluate the outcomes of one- and two-piece mushroom keratoplasty configurations for various corneal pathologies. MATERIAL AND METHODS: A total of 32 patients (32 eyes) with corneal perforations, descemetocele and deep corneal opacities underwent surgery. Depending on the surgery technique, the patients were divided into 2 groups: group I (17 eyes) underwent manual one-piece mushroom keratoplasty according to our own technique; group II (15 eyes) underwent modified two-piece microkeratome-assisted mushroom keratoplasty according to the technique by M. Busin. All patients underwent clinical and functional studies before surgery and in the course of a 1-year follow-up. RESULTS: Transparent engraftment of the cornea was achieved in 82 and 80% of cases; best corrected visual acuity (BCVA) averaged 0.57±0.19 and 0.53±0.17; spherical component of refraction varied from 1.25 to +5.5 and from 1.25 to 6.0 diopters; mean corneal astigmatism was 3.15±1.73 and 3.21±1.89 diopters in groups I and II, respectively. At 6 months after surgery, the mean endothelial cell density (ECD) was 2336±198 and 2291±175 cells/mm2, at 1 year - 2041±189 and 1955±161 cells/mm2 in groups I and II, respectively. CONCLUSION: One- and two-piece mushroom keratoplasty is effective in surgical treatment of various corneal pathologies ensuring a high rate of transparent corneal engraftment. The revealed risk of false chambers formation between separate parts of the graft after the two-piece method could in most cases be eliminated by re-injecting air into the anterior eye chamber.

[Diagnosis and treatment of adenoviral keratoconjunctivitis]. / Diagnostika i lechenie adenovirusnogo keratokon"yunktivita.

PURPOSE: To improve the treatment of adenoviral lesions of the eye based on express diagnostics by the fluorescent antibody technique (FAT) and the use of modern drugs. MATERIAL AND METHODS: The study included 184 patients (333 eyes) with various manifestations of adenoviral lesions of the ocular surface, who were divided into two groups: group 1 (149 patients, 196 eyes) - acute form, and group 2 (76 patients, 137 eyes) - long lasting form. Effectiveness of the proposed treatment was evaluated against separate group 3 (controls) consisting of 28 people (46 eyes) with completed acute adenovirus infection, who had previously received antibiotic and corticosteroid therapy in other clinics. Conjunctival scrapings of study patients were examined with FAT in our proposed modification. Study patients received local therapy with modern drugs (Okomistin, Aktipol). RESULTS: FAT detected the adenovirus antigen in 169 cases in group 1 (86%) and in 99 cases in group 2 (72%). Treatment duration amounted to 12±6 days in group 1, 18±8 days in group 2, and 29±7 days in controls. In both study groups, the duration of treatment was significantly reduced in comparison with the controls (p<0.01). Stable clinical effect and complete restoration of visual acuity have been achieved in most cases. There were no allergic and side effects from the therapy. CONCLUSION: Fluorescent antibody technique is a fast and effective way to diagnose adenovirus infection in ophthalmology. In terms of therapy, the use of an antiseptic, an antiviral drug and diluted corticosteroids is the most rational approach.

[Results of the application of complex physiotherapeutic neurostimulation in optical neuropathies of various genesis]. / Rezul'taty primeneniya kompleksnoi fizioterapevticheskoi neirostimulyatsii pri opticheskikh neiropatiyakh sosudistogo geneza.

Optical neuropathies (ON) are the outcome of many diseases of various origins. The main ones are classified as inflammatory, vascular and traumatic ON. ON lead to subatrophy of the optic nerve, but even after the completion of treatment, it is possible to improve visual functions by using physiotherapeutic means of ophthalmic rehabilitation. OBJECTIVE: To evaluate the effectiveness of complex physiotherapeutic neuro-ophthalmostimulation in case of ON of vascular origin. MATERIAL AND METHODS: The study included 60 patients (120 eyes) with a verified diagnosis of optic neuropathy of vascular origin, who were divided into 2 groups comparable in age, gender and anatomical and functional characteristics: the main group of 30 patients (60 eyes) and the control group - 30 patients (60 eyes), including 24 men and 36 women, mean age was 66.2±4.1 years, disease duration was 4.1±1.7 years. All patients underwent courses of conservative treatment with vitamins according to the available ophthalmological standards, repeating them 1-2 times a year, the last of which was six months before the present study. Physiotherapy courses and patients did not pass. 20 healthy volunteers were taken to create basic indicators of the «norm¼ of the applied research methods. Patients of the main group used a set of procedures: transcranial magnetic electrical stimulation, endonasal electrophoresis with the drug neuroprotector Semax 0.1%, oxybaric chamber and acupuncture. Patients in the control group were prescribed basic therapy, including taking the vitamin complex BEROCCA for 3 months. Breakpoints: before treatment and at times: 1st week, 12 weeks and 24 weeks after the course of treatment, according to the standard recommendations for international multicenter studies. The following were assessed: visual field boundaries (dilation meridians; in deg.), light sensitivity (MS, MD; in dB), indicators of the state of the retinal ganglion layer (GCS thickness, volume loss): Avg CCG (in µm), FLV, GLV (in %). RESULTS: When evaluating the results in patients of the main group who received complex neurostimulation, the therapeutic efficacy in a week after the end of treatment was 94%, in 12 weeks - in 88% and in 24 weeks - 83%, while in patients of the control group for all studied indicators showed only a positive trend and therapeutic efficacy did not exceed 30-42%. CONCLUSION: Under the influence of the developed neurostimulating complex, the activity of nerve cells objectively increases, leading to a significant increase in the boundaries of the field of view and light sensitivity and a decrease in global losses of the retinal ganglion complex and optic nerve.

[Differential diagnosis of chylothorax in therapeutic practice].

The article is devoted to the differential diagnosis of chylous pleural effusion. The spectrum of traumatic and non-traumatic causes of chylothorax is discussed in detail. Examples of necessary diagnostic measures are given for the accumulation of milky fluid in the pleural cavity in order to verify chylothorax.

[Anterior eye segment damage in coronavirus infection (COVID-19)]. / Porazhenie perednego segmenta glaza pri koronavirusnoi infektsii (COVID-19).

Conjunctivitis may appear as the first symptom of the coronavirus infection (COVID-19). In isolated cases, the lesion of the conjunctiva evokes a systemic infectious process. Currently, the conjunctiva is not considered as an area of long-term reproduction of coronavirus, and its damage is caused by hyperproduction of pro-inflammatory cytokines (especially IL-6); development of iridocyclitis and keratoconjunctivitis is also possible. Most often, local corticosteroids are used to treat these processes, although their use requires caution due to the risk of activating secondary infection (herpetic bacterial, fungal), which often develops as a result of immunodeficiency caused both by COVID-19 and the massive corticosteroid and antibiotic therapy employed when the course of the disease is severe. The severe condition of patients, the lung ventilation, and the prone position all contribute to corneal erosions, exposure keratopathy, pseudomonas aeruginosa keratitis and angle-closure glaucoma attacks. The risk of transmission of coronavirus infection during keratoplasty is estimated as minimal.

Genetic diversity of enteroaggregative Escherichia coli.

Studied 74 E. coli strains isolated from stool samples (60) and urine samples (14) of patients examined for clinical indications. Molecular methods included: PCR with electrophoretic detection of genes associated with diarrheal E. coli pathogroup EAgEC (aggR, aaf, aap, aatA, pet, ast, aai) and ExPEC pathogroup UPEC (pap, sfa, afa, kpsMT II, iutA, hlyA, cnf), MLST typing, whole genome sequencing. Strains isolated from stool samples were significantly more likely (88.3%, p > 0.05) to be typical EAgECaggR+ compared to atypical EAgECaggR-. Strains isolated from urine samples, significant differences between typical and atypical EAgEC were not detected (p > 0.05). Genes associated with ExPEC were present in all strains isolated from urine samples and in 45 strains (75%) isolated from stool samples. Coproisolates belonged to 10 serogroups and 13 serovars: O3:H2, O11:H10, O16:H48, O51:H30, O55:H21, O73:H18, O73:H33, O86:H2, O86:H10, O92:H33, O140:H2, O159:H10. Two strains had unique nucleotide sequences of genes encoding O-antigens that were missing from the SerotypeFinder database. 80% of EAgEC isolated from feces and urine was characterized by an enteroaggregative/uropathogenic genotype (EAgEC/UPEC). Most of the strains isolated from urine belonged to the virulent clone of high-risk epidemic spread ST 38 associated with hybrid strains of UPEC / EAgEC.

[Phylogenetic groups and virulence genes of Escherichia coli strains isolated from the children gut microbiota.]

Escherichia coli is characterized by a wide intraspecific diversity. The species includes both commensals and pathogens that cause diarrhea and extra-intestinal diseases. Pathogenic strains differ from non-pathogenic ones by the presence of virulence factors and their genes. The phylogenetic structure of the species is represented by four main groups (A, B1, B2, D), which differ in their prevalence among residents of different geographical regions. Pathogenic members of the species have been studied in detail, while non-pathogenic strains have not received such attention. This report presents the results of a study of 511 E. coli strains isolated from the gut microbiota of children without diarrhea and urinary tract infections, aged from 1 month to 17 years, living in St. Petersburg. The main phylogenetic groups were determined by PCR, and E. coli virulence genes associated with diarrhea and extra-intestinal diseases were identified. Results: population structure of E. coli is represented by the following groups: A-33.3%, B1-6.7%, B2-34.0%, D-26%. In the studied population 2.5% of strains belonded to EPEC and 4.5% to EAggEC. EPEC virulence genes were more often detected in strains of phylogroup B1, and EAggEC virulence genes in isolates of phylogroup D. The prevalence of extra - intestinal virulence genes was as follows: pap - 29.5%; sfa - 19.8%; afa - 3.3%; hly - 20.9%; cnf - 17.4%; aer-20.0%. The pap, sfa, hly, and cnf genes were detected mostly in the B2 phylogenetic group. Obtained data shows the similarity of E. coli phylogenetic groups structure in St. Petersburg with E. coli populations isolated from residents of Paris and Sydney. Analysis of the virulence genes prevalence showed the dependence of their presence on the genetic background bacteria.

[Laboratory errors for identification of the escherichia coli strains of the serological groups O6 and O25 as acute intestinal infections.]

Were studied the genes encoding the virulence factors of 221 strains: E. coli O6:H1 (194) and E. coli O25:H4 (27), isolated in 2014-2018 from stool samples of children and adults examined according to epidemic indications. Molecular methods included PCR with hybridization-fluorescence and electrophoresis detection of amplified products. The strains did not have virulence genes for diarrheagenic E. coli (DEC) pathogroups EPEC, ETEC, EIEC, EHEC, EAggEC, and belonged to the phylogenetic group B2. They contained from four to eight genes encoding virulence factors of ExPEC: E. coli O6:H1 - pap (68,6%), sfa (87,6%), fimH (96,4%), hly (62,4%), cnf (74,7%), iutA (97,9%), fyuA (95,9%), chu (100%); E. coli O25:H4 - pap (66,7%), afa (22,2%), fimH (100%), hly (44,4%), cnf (44,4%), iutA (100%) , fyuA (100%), chu (100%). The antimicrobial susceptibility testing to 6 classes of antimicrobials (beta-lactams, fluoroquinolones, aminoglycosides, nitrofurantoin, sulfanilamide, trimethoprim / sulfamethoxazole) according the EUCAST. 60,3% of E. coli O6:H1 were sensitive to antibiotics, E. coli O25:H4 remained sensitive to carbapenems and nitrofurans. Extended-spectrum cephalosporins resistance was due to the production ESBL (CTX-M). The 57,1% resistant strains of E. coli O6:H1 and 100% of E. coli O25:H4 strains belonged to the MDR phenotype. The XDR phenotype had one in five MDR strains of E. coli O6:H1 and E. coli O25:H4. All strains of E. coli O25:H4 belonged to ST131. Given the important role of E. coli in human pathology, detection of virulence genes should be performed to confirm the etiological significance of the isolated strain.

[Methods of preserving donor corneas]. / Metody konservatsii donorskikh rogovits.

Corneal transplantation is the most successful and frequently performed allotransplantation procedure. Benign outcome of penetrating and posterior keratoplasty depends on viability of the corneal tissue and presence of the proper endothelial layer, which retaining is one of the main objectives of preservation of donor material. Methods designed to reach this goal may be classified according to storage duration as short-term (storage in a moist chamber), medium-term (hypothermia), long-term (organ cultivation) and unlimited (cryopreservation). While there are reports of successful application of cryo-conserved corneas for penetrating keratoplasty, its use for this type of transplantation is limited due to complexity of the method and very high requirements for the quality of donor material. However, being a method of unlimited storage, it still attracts the attention of researchers. Taking into account the ongoing development of innovative keratoplasty technologies and the tendency for predominant use of layered grafts that in some cases do not need viable endothelium, methods of preservation of such material for anterior lamellar and intralamellar keratoplasties become relevant. In this context, cryopreservation without the use of cryoprotective agents is promising because it allows simple and fast accumulation of large amount of material with long storage time. Further research on the development of preservation methods of the corneal endothelial layer at sub-zero temperatures may allow storing the material by cryopreservation that will be suitable for penetrating and posterior keratoplasty.

[Molecular-genetic characterization of shiga-toxin producing Escherichia coli isolated during a food-borne outbreak in St. Petersburg in 2013].

UNLABELLED: Shiga toxin-producing Escherichia coli (STEC) food-borne infections are reported worldwide and represent a serious problem for public healthcare. In the Russian Federation there is little information on epidemiology and etiology of STEC-infections as well as on molecular-genetic peculiarities of STEC pathogens. OBJECTIVE: Our aim was to describe a food-borne outbreak as hemorrhagic colitis (HC) along with hemolytic uremic syndrome (HUS), enterocolitis, and acute gastroenteritis in children in St. Petersburg in 2013. METHODS: Epidemiological, microbiological, molecular-genetic and bioinformatic methods were applied. RESULTS: Objects to study were clinical specimens, milk and food samples, as well as STEC strains isolated during the outbreak. The outbreak of food-borne infection was found to be caused by STEC-contaminated raw milk as confirmed by epidemiological analysis, detection of STEC DNA and isolation of relevant pathogens in milk and sick children fecal specimens. The whole-genome sequencing revealed two groups ofpathogens, E. coli O157:H7 and E. coli O101:H33 among collected strains. Group I strains were attributed to the previously known sequence type ST24, while group II strains belonged to the previously non-described sequence type ST145. In strain genomes of both groups there were identified nucleotide sequences of VT2-like prophage carrying stx2c gene, plasmid enterohemolysin gene, and gene of the STEC main adhesion factor intimin. Gene of intimin gamma was identified in E. coli O157:H7 strains and intimin iota 2 in E. coli O101:H33 strains. The latter previously was identified only in enteropathogenic E. coli (EPEC) strains. CONCLUSION: The additional knowledge of epidemiology and biology of STEC pathogens would assist clinicians and epidemiologists in diagnosing, treating and preventing hemorrhagic colitis.

[Characteristics of enterohemorrhagic Escherichia coli O145:H28 isolated from a patient with hemolytic-uremic syndrome].

AIM: Determine etiologic significance of clinical strains of E. coli O145:H28 isolated from feces of a patient with hemolytic-uremic syndrome (HUS). MATERIALS AND METHODS: 20 E. coli strains isolated from feces of a patient with HUS that developed after an acute intestine infection were studied. Antigenic structure of strains was determined by sequencing of rib and fliC genes; presence of virulence genes (pap, aaf, sfa, afa, eaeA, bfpA, ial, hly, cnf, stx1, stx2, LT, ST and aer)--in PCR; ESBL production --by double disk method, ESBL genes--in PCR. RESULTS: The strains contained rfb gene coding O145, fliC gene coding H7. Genes coding synthesis of stx2-toxin and intimin (eaeA) were detected. The strains were resistant to beta-lactams due to production of CTX-M class ESBL. CONCLUSION: A causative agent E. coli O145:H28 was isolated from a patient with HUS that produces stx2-toxin and CTX-M class ESBL and has not been previously registered in Russian Federation.

[Prevalence and risk factors of delirium in COVID-19]. / Rasprostranennost' i faktory riska deliriya pri COVID-19.

OBJECTIVE: To assess the frequency and risk factors of delirium in patients hospitalized with COVID-19. MATERIAL AND METHODS: Four hundred and forty patients admitted to the Infectious Diseases Hospital of the University Clinical Hospital No.3 of Sechenov University were included in the study. The Patient Health Questionnaire (PHQ-9), the Generalized Anxiety Disorder Screening Questionnaire-7 (GAD-7), the Insomnia Severity Index (ISI), the Delirium severity rating scale (DRS-R-98) were administered. RESULTS: Delirium was detected in 27.8% of patients. Significant risk factors were age (p=0.002), severity of respiratory failure (p=0.005), concomitant somatic disease (p=0.003), and respiratory therapy (p<0.001). There was an association between severe anxiety (p<0.001) and insomnia (p=0.07) observed at admission with the risk of developing delirium during the hospital stay. CONCLUSION: The study reveals a high prevalence of delirium in patients with COVID-19. In order to prevent delirium and/or reduce the risk, early diagnosis and identification of preclinical forms are of particular importance.

Characterization of the C6D7-RBD Human Monoclonal Antibody Specific to the SARS-CoV-2 S Protein Receptor-Binding Domain.

The new coronavirus infection COVID-19 is an acute viral disease that affects primarily the upper respiratory tract. The etiological agent of COVID-19 is the SARS-CoV-2 RNA virus (Coronaviridae family, Betacoronavirus genus, Sarbecovirus subgenus). We have developed a high-affinity human monoclonal antibody, called C6D7-RBD, which is specific to the S protein receptor-binding domain (RBD) from the SARS-CoV-2 Wuhan-Hu-1 strain and exhibits virus-neutralizing activity in a test with recombinant antigens: angiotensin-converting enzyme 2 (ACE2) and RBD.

[The characteristics of biological properties of E. coli O104:H4--the causative agent of large-scale alimentary ictus in Germany may 2001].

The review presents the characteristics of E. coli O104:H4, the causative agent of large-scale alimentary ictus in Germany in spring time 2011. The antigenic characteristics and factors of E. coli pathogenicity are taken into account. The causative agent has a combination of pathogenic factors of two groups of diarrheigenic Escherichia: shigella similar toxin, specific for entero-hemorrhagic E. coli and adhesins of enteroaggregative E. coli.

[Virulence markers of Escherichia coli O1 strains].

AIM: To detect virulence genes in clinical isolates of Escherichia coli O1 using polymerase chain reaction (PCR). MATERIALS AND METHODS: One hundred and twenty strains of E.coli O1 strains isolated from faeces of patients with acute diarrhea (n = 45) and healthy persons (n = 75) were studied. PCR with primers for rfb and fliC genes, which control synthesis of O- and H- antigens respectively, was used. Fourteen virulence genes (pap, aaf, sfa, afa, eaeA, bfpA, ial, hly, cnf, stx1, stx2, lt, st, and aer) were detected by PCR primers. K1-antigen was determined by Pastorex Meningo B/E. coli O1 kit (Bio-Rad). RESULTS: rfb gene controlling O-antigen synthesis in serogroup O1 as well as fliC gene controlling synthesis of H7 and K1 antigens were detected in all strains. Thus all E. coli strains had antigenic structure O1:K1 :H-:F7. Virulence genes aafl, sfa, afa, eaeA, bfpA, ial, hly, cnf, stx1, stx2, lt, and st were not detected. All strains owned pap and aer genes regardless of the presence of acute diarrhea symptoms. CONCLUSION: It was shown that E. coli O1:KI:H-:F7 strains do not have virulence genes which are characteristic for diarrhea-causing Escherichia. In accordance with the presence of pap and aer genes they could be attributed to uropathogenic Escherichia (UPEC) or avian-pathogenic Escherichia (APEC). It is necessary to detect virulence factors in order to determine E. coli as a cause of intestinal infection.

[Unipolar depression: a gender-based comparative analysis]. / Depressivnye rasstroistva s monopolyarnym techeniem: sravnitel'nyi analiz po gendernomu priznaku.

OBJECTIVE: To compare socio-demographic and clinical characteristics in men and women with depression. MATERIAL AND METHODS: We studied 388 patients with unipolar depression, 74 men and 314 women. The survey was carried out using the clinical-psychopathological method and psychometric scales for depression. RESULTS AND CONCLUSION: Gender differences are identified for several parameters. Women are older, more often had comorbid somatic and organic disorders; men have personality disorders and chronic depression. The severity of depression in women was higher due to symptoms such as reported sadness, anxiety, reduced sleep and appetite, concentration difficulties, and suicidal thoughts. Apparent sadness did not significantly differ in men and women. Anhedonia was more pronounced in men.

[Recurrent herpetic corneal erosion: diagnosis and treatment].

Recurrent corneal erosion (CE) is a common anterior eye disease, which usually occurs after injury, substantially limits a patient's ability to work, and is intractable. The authors single out an individual form of CE herpetic CE (HCE) on the basis of immunofluorescence assay of a conjunctival scrape, which shows the high concentration of herpes simplex virus antigen in 53% of CE cases. Confocal microscopy revealed epithelocyte polymorphism and basement membrane defects. The treatment of patients with HCE involved 2 steps: 1) relief of acute signs of the disease via drug therapy, rapid local autocyte cinotherapy (RLACCT), or phototherapeutic keratectomy (PTK) and 2) prevention of recurrences. For this the authors developed a method based on the systemic use of a new composition of the intradermal herpes vaccine Vitaherpavac in combination with the subcutaneous interferon inducer Poludan. RLACCT was found to be the most effective medical treatment for CE and PTK was the most effective surgical one. Vaccination with the concurrent subcutaneous injection of Poludan is an effective method in preventing recurrent HCE. During a follow-up of 2 years or more, 81% of the patients achieved clinical resolution; there was a decrease in recurrence rates and severity in 15.1% and no effect in 3.8%.