The using of the polymerase chain reaction for the detection of resistance genes in gram-negative bacteria in routine practice in a pediatric hospital.

Objective - assessment of RT-PCR for the detection of carbapenem-resistance genes in gram-negative bacteria. A total, 499 strains of gram-negative microorganisms isolated in two pediatric hospitals in 2019-2020 were studied. Species identification was performed using MALDI-ToF mass-spectrometry (Bruker Daltonics, Germany). Meropenem and imipenem minimal inhibitory concentration (MIC) was determined by E-test method (BioMerieux, France). The presence of acquired carbapenemase genes of IMP, NDM, VIM, KPC, OXA-48, OXA-23, OXA-40, OXA-58-groups was determined by RT-PCR. Klebsiella pneumoniae (34%), Escherichia coli (4%), Serratia marcescens (6%) and other members of Enterobacterales (6%), also gram-negative non-glucose-fermenting bacteria Acinetobacter baumannii (14%), Pseudomonas aeruginosa (36%) were found among selected strains. Carbapenemase production was found in 385 isolates (77%). The main mechanism determining carbapenem resistance in P. aeruginosa was the production of blaVIM (100%). A. baumanii strains harbored OXA-23 (55%) and OXA-40 (45%) carbapenemases. The major determinant of carbapenem resistance in K. pneumoniae isolates was OXA-48 carbapenemase, detected in 63% strains, 13% of the strains possessed blaNDM-group, 16% isolates had a combination of blaNDM-group and blaOXA-48-like. Carbapenemase of KPC-group was found in 8% K. pneumoniae strains. OXA-48 carbapenemase prevailed (95%) among S. marcescens strains. Most of E. coli isolates harbored metallo-beta-lactamase NDM (89%). Other members of Enterobacterales most often had OXA-48 carbapenemase (57%), 39% of the isolates carried blaNDM-group. In one strain, a combination of blaNDM-group and blaOXA-48-like was discovered. RT-PCR is a fast and reliable method for the detection of acquired carbapenemases and can be recommended for routine use in bacteriological laboratories.

[Clinical and sociodemographic characteristics of patients with the first depressive episode and recurrent depression]. / Klinicheskaya i sotsiodemograficheskaya kharakteristika bol'nykh s pervym depressivnym epizodom i rekurrentnoi depressiei.

OBJECTIVE: To comparare socio-demographic and clinical characteristics of patients with the first depressive episode and recurrent depression. MATERIAL AND METHODS: Three hundred and twenty one patients with unipolar depression, including 96 patients with first depressive episode and 225 patients with recurrent depression, were examined using clinical and psychometric methods. RESULTS AND CONCLUSION: There were differences in clinical characteristics between groups but such factors as gender, marital status, level of education, family history of mental disorders and personality were similar. With each new episode of recurrent depression, the next episode tends to be more severe with more intense pessimistic and suicidal thoughts but fewer anxiety and complaints of depressive mood that affects the differences and requires further research, especially considering the effect of therapy.

Emergence of a ST307 clone carrying a novel insertion element MITEKpn1 in the mgrB gene among carbapenem-resistant Klebsiella pneumoniae from Moscow, Russia.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) represents a major nosocomial pathogen with only a few antimicrobial agents, including colistin, remaining active. However, the emergence of colistin-resistant (Col-R) isolates is compromising the activity of colistin. In this study, a collection of 159 CRKP recovered from three hospitals in Moscow (Russian Federation) was examined. The isolates demonstrated resistance to cephalosporins (100%), ciprofloxacin (92.5%), fosfomycin (90.1%), netilmicin (81.1%), gentamicin (84.3%) and amikacin (49.7%). The rate of colistin resistance (MIC > 2 mg/L by broth microdilution) was 44.7%; moreover, 6.7% of isolates were tigecycline-resistant. Among 18 sequence types (STs) discovered, isolates of five lineages including ST307 (n = 46; 28.9%), ST395 (n = 40; 25.2%), ST377 (n = 17; 10.7%), ST48 (n = 17; 10.7%) and ST23 (n = 16; 10.1%) dominated. Carriage of a blaOXA-48-like carbapenemase gene was detected in 146 CRKP (91.8%); 11 (6.9%) and 2 (1.3%) isolates harboured blaNDM-1 and blaKPC-3, respectively. Among 71 Col-R isolates, colistin MICs ranged from 4 mg/L to >1024 mg/L (MIC50/90, 2/512 mg/L). All Col-R isolates were mcr-1-negative. In 19 (26.8%) Col-R isolates, inactivation of mgrB by insertion sequences IS1A, IS1R, ISKpn14 and ISKpn26 and a novel miniature inverted-repeat transposable element (MITE) Kpn1 was observed. Carriage of MITEKpn1 was restricted to six ST307 isolates and affected mgrB at nucleotide position 75. mgrB deletion was observed in four (5.6%) Col-R isolates. Moreover, PmrA and/or PmrB were altered in three (4.5%) Col-R isolates with wild-type mgrB. Thus, blaOXA-48-like-carrying Col-R ST307 K. pneumoniae is emerging as a dominant clone in Moscow.

[Integron diversity in blavim-2-carrying carbapenem-resistant clinical Pseudomonas aeruginosa isolates.]

The growing prevalence of metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa in nosocomial pathogen populations has been attributed to their clonal spread, and/or horizontal transfer of MBL determinants in mobile genetic elements, including integrons. To characterize the genetic background of the beta-lactamase VIM-2 encoding gene in the population of carbapenemresistant (Carba-R) P. aeruginosa clinical isolates.The detection of class 1 integrons was performed by PCR. Typing of the class 1 integrons containing the blaVIM gene cassette was performed by the PCR-restriction fragment length polymorphism (RFLP) approach followed by sequencing of variable regions of class 1 integrons. Five types of the blaVIM-2-carrying integrons were identified: ST654-isolates accounting for more than 50% of the Carba-R population harbored In56; ST235-isolates contained In559 (26% Carba-R isolates); ST111-isolates (19% Carba-R isolates) were characterized by carrying In59-like integron; two ST235-isolates harbored In59 and In249 each. Except In56, carrying the only blaVIM-2-gene cassette, all other identified integron types harbored the genes of resistance to trimethoprim and/or aminoglycosides. No new types of integrons were identified in the P. aeruginosa clinical isolates. The observed correlation of the integron type with specific STs indicates a clonal dissemination of significant resistance determinant producers - ST111, ST654 and ST235 epidemic lines. The features of the integron variable regions can be used for the epidemiological characterization of clinical P. aeruginosa isolates.

[The comparison of methods for determination of colistin susceptibility in carbapenem-resistant Klebsiella pneumoniae.]

In recent years, because of carbapenemase spreading in Klebsiella pneumoniae strains, the antibiotic of reserve group, colistin, is increasingly prescribed. In vitro testing of colistin susceptibility in everyday practice has a number of difficulties due to the cationic properties of molecule and weak diffusion into agar. Therefore it is recommended to use the reference Broth Microdilution Method (BMD) for determination of the Minimum Inhibitory Concentration (MIC) for colistin. The purpose of the study was to determine susceptibility to colistin in 119 carbapenem-resistant K. pneumoniae (CRKp) which were isolated from the patients at three hospitals in Moscow in 2012-2016 by the broth microdilution method (BMD) and to compare these data with the ones obtained by epsilometer test (E-test) and VITEK 2 Compact. The proportion of resistant isolates (MIC>2 mg/L) was 52%, 39%, 35% respectively. Both commercial methods demonstrated a high level of the very major error (VME) that was 26% for the E-test method and 34% for the VITEK 2 Compact. The values of categorical agreement and essential agreement (CA, EA) were less than 90%. A single major error (ME) was detected for the VITEK 2 Compact. In conclusion, results of both commercial tests for determination of MIC for colistin showed differences with the results of the reference BMD. It is necessary for clinical laboratories to be aware about this discrepancy and to use E-tests and VITEK 2 Compact with caution to determine colistin susceptibility.

Bilateral thalamic stroke in patient with patent foramen ovale and hereditary thrombophilia.

Patent foramen ovale and hereditary thrombophilia are both known risk factors for ischemic stroke. Artery of Percheron is a rare anatomical variant in which vast areas of the midbrain and thalamus have a single source of blood supply. This case report presents a 45-years old female patient with bilateral thalamic stroke due to Percheron artery occlusion, with a combination of hereditary thrombophilia and patent foramen ovale as the risk factors. Modern approaches to the diagnosis and secondary prevention of this pathology are also discussed herein.

[SPECTRUM OF ANTIBIOTIC RESISTANCE AND PREVALENCE OF OXA-CARBAPENEMASES AMONG ACINETOBACTER BAUMANNII STRAINS, ISOLATED FROM PATIENTS OF SURGICAL AND REANIMATION DEPARTMENTS IN MOSCOW].

AIM: Characterize spectrum of antibiotics resistance of Acinetobacter baumannii strains, isolated from patients of 8 surgical and reanimation departments of 3 medical institution of Moscow, and determine molecular-genetic mechanisms of stability of their carbapenem-resistant forms. MATERIALS AND METHODS: 95 strains of A. baumannii, isolated from patients of reanimation and surgical departments of Moscow in 2012-2014, were studied. Sensitivity of strains to antibiotics was tested phenotypically according to recommendations of EUCAST. The presence of VIM, IMP, OXA-23, OXA-40, OXA-48, OXA-58 and NDM genes in the studied strains was determined by polymerase chain reaction in real time. RESULTS: 86.3% of strains turnedout to be non-sensitive to carbapenems, sensitive--13.7%. 80.0% of strains were non-sensitive to gentamicin, 80.0% of strains--to netilmicin, 94.7% of strains--to ciprofloxacin 2.1%--to colistin. 91.6% of isolates have shown non-sensitivity to members of 2 and more classes of antibiotics, 78.9% of strains--to members of 3 classes. 2 strains were panresistant, 4.2% (4/95) of the isolates were sensitive to all the classes of antibiotics. Metallo-ß-lactamases were not detected. Genes of carbapenemases (OXA-23 and/or OXA-40) were detected in 85.3% (81/95) of strains, characterized phenotypically as non-sensitive to carbapenems. CONCLUSION: The results obtained shown an increase of resistance to carbapenems and multiple resistance in clinically significant strains of A. baumannii. Resistance to carbapenems is associated with OXA-23 and OXA-40 genes. The conclusions allow to justify perspectives of introduction of technologies of molecular-genetic testing of antibiotics resistance.

Macrolide Resistance and Its Molecular Genetic Mechanisms in Streptococcus pyogenes Isolated from Children.

The frequency and mechanisms of resistance to macrolides in Streptococcus.pyogenes isolated within 3 periods: 2011-2012 (246 strains), 2013-2014 (273 strains) and from January to November of 2015 (120 strains) were studied. The strains of S.pyogenes (639) were isolated from 17107 nasopharyngeal, vaginal and middle ear discharge smears of children on their visits to physiciants or hospitalization at somatic hospital departments. The susceptibility was tested by the disk diffusion method and E-test strips. Identification of the mechanisms of resistance to macrolides and lincosamides included phenotypic and molecular genetic methods. PCR was used to determine ermB and mef genes in 23 erythromycin resistant isolates. As compared to 2011-2012, resistance of S.pyogenes to macrolides increased from 5 to 16% in 2015 and that to clindamycin from 2 to 10%. Among 23 erythromycin resistant strains 6 (26.1%) belonged to the M phenotype, 3 (13.0%) belonged to the iMLS(b) phenotype and 14 (60.9%) belonged to the cMLS(b) pheno-type. The results of detecting the macrolide resistance genes in S.pyogenes showed that only 26.1% of the isolates expressed the mefA gene. The predominant share (65.2%) of the erythromycin resistant isolates possesed the ermB gene as a determinant and in 4.3% of the isolates the ermB gene was associatied with the mefgene. No resistance genes were detected 1 isolate. Therefore, the main mech- anism that determined resistance of S.pyogenes to macrolides was methylation of ribosomes mediated by the ermB gene.

[Antibiotic Resistance and Its Molecular Mechanisms in Carbapenem-Nonsusceptible Klebsiellapneumoniae Isolated in Pediatric ICUs in Moscow.]

Klebsiellapneumoniae is a significant pathogen associated with hospital infections. Its was isolated in intensive care units (ICU) at two pediatric hospitals in Moscow in 2012-2014 from 41% (387/935) of the patients. The rate of carbapenem-nonsusceptibility (Carba-NS) amounted to 25% for imipenem and 27% for meropenem. For further analyses, 67 isolates were selected, including 57 Carba-NS and 10 Carba-susceptible (Carba-S). Among the isolates, 100% was nonsusceptible to the III-IV generation cephalosporins, 50-84% was resistant to aminoglycosides. The rate of nonsusceptibility to ciprofloxacin and phosphomycin exceeded 90%. All the tested Carba-S Kpneumoniae isolates were susceptible to tigecycline, whereas 25% of the Carba-NS isolates was tigecycline-NS. The prevalence of the colistin-NS isolates was the same in Carba-S (20%) and Carba-NS (26%) bacteria. The blamrx_ gene was carried by 100% of the Carba-S isolates, combining with the blaTEM gene in 60% of the isolates. In 89% of the Carba-NS isolates the OXA-48 carbapenemase was detected, which was combined with CTX-M and/or TEM in all but 1 isolate. Thus, over the last decade, the rate of Carba-NS among nosocomial Kpneuynoniae increased and the OXA-48 carbapenemase was shown to be dominating in the mechanism of Carba-NS in the pediatric ICUs in Moscow.

[BIOFILM FORMATION BY STREPTOCOCCUS PNEUMONIAE].

The biofilm process in Streptococcus pneumoniae (pneumococcus) is described. Virtually all wild-type pneumococci are capable of the biofilm formation. The pneumococcal capsule may reduce the biofilm production, and the propensity to form biofilms has a reverse correlation with the amount of the capsule material. Invasive pneumococcal isolates and noninvasive strains that persist in the nasopharynx have different biofilm potential. A number of issues related to effector and regulatory factors in the pneumococcal biofilms are discussed in this review. In the summary, a biofilm may be essential only for the persistent pneumococcal infection.

[Serotype diversity and antimicrobial resistance of Streptococcus pneumoniae].

The pneumococcus (Streptococcus pneumoniae) is a common bacterial pathogen responsible for various infections, especially in children below 5 years of age. The severity of pneumococcal infections varies from self-limiting mucosal infections, including acute otitis media, sinusitis, and noninvasive pneumonia, to life-threatening invasive disease like bacteremia and meningitis. A high incidence of pneumococcal infections is combined with a constantly growing antibiotic resistance of this pathogen. The growing resistance is thought to be associated with misuse of antibiotics and emerging of resistant clones that may spread throughout the entire population. Pneumococcal polysaccharide conjugate vaccines (PCV) contain an assortment of pneumococcal capsular polysaccharides (from 7 to 13) that produce serotype-specific protective antibodies. Since early 2000's, the introduction of PCV into national immunization programs has been shown to substantially decrease the incidence of invasive pneumococcal disease and pneumococcal carriage associated with vaccine-type pneumococci in many countries. In 2014, PCV vaccination was included in the Russian national calendar of prophylactic vaccination. The present article reviews the current literature on serotype prevalence, antibiotic susceptibility, and PCV effect on the evolution of pneumococcus.

[Monitoring and antibiotic resistance profile of tracheal aspirate microbiota in ICU children with severe craniocerebral trauma].

Nosocomial infections and their rational antibiotic treatment represent a major challenge for the healthcare nowadays. In this context, gramnegative bacteria including Pseudomonas aeruginosa, Acinetobacter baumanii and Enterobacteriaceae spp. are etiologically important and characterized by a significant level of antibiotic resistance. To examine dynamics of the respiratory tract colonization by hospital flora, tracheal aspirates obtained at three time points from 69 children with severe craniocerebral trauma during their stay in ICU were analysed. Colonization was observed on the 4th day of the ICU stay with predomination of K. pneumoniae (45%) and A. baumanii (27-37%). P. aeruginosa was detected after the 8th day of the ICU stay with the isolation rate of 33%. Substantial proportions of P. aeruginosa (61%), A. baumanii (78%) and K. pneumoniae (25%) were resistant to carbapenems. In 65 carbapemen resistant isolates, the presence of carbapenemases was examined using PCRs. OXA-48 carbapenemase was detected in 11 out of 14 (78%) K. pneumoniae isolates. Among the A. baumanii isolates, 30/31 (97%) carried OXA-40 and 1/31 (3%) had OXA-23 carbapenemases. None of the examined A. baumanii and K. pneumoniae isolates produced metallo-betalactamases (MBL). In contrast, all 20 carbapenem resistant P. aeruginosa isolates produced a MBL, and in 12 out of 20 (60%) of theme VIM-2 was detected. Thus, gramnegative nosocomial microflora rapidly colonizes ICU patients and has a high level of resistance to antibiotics, including carbapenems.

[Acinetobacter: microbiological, pathogenetic and resistant properties].

Species of the genus Acinetobacter represent opportunistic bacteria with a growing clinical significance. In this literature review, we focus on the current role of Acinetobacter in infectious pathology and describe physiology, taxonomy, ecology, pathogenicity, and antibiotic resistance of these bacteria. Molecular pathogenesis and regulation of virulence factors in Acinetobacter spp. are described in detail. The majority of acinetobacterial infections are associated with A. baumannii and occur predominantly in an immunocompromised host. Usually, acinetobacterial infections are characterized by local purulent inflammation; in severe cases, meningitis and sepsis may develop. Antibiotic resistance ofAcinetobacter is a major clinical problem; therefore we give special attention to laboratory testing of resistance as well as identification of Acinetobacter. In addition, treatment and prophylaxis of acinetobacterial infections are discussed.

Palmitic and stearic acids bind Ca2+ with high affinity and form nonspecific channels in black-lipid membranes. Possible relation to Ca2+-activated mitochondrial pores.

A mitochondrial hydrophobic component that forms Ca2+-induced nonspecific ion channels in black-lipid membranes (Mironova et al., 1997) has been purified and its nature elucidated. It consists of long-chain saturated fatty acids--mainly palmitic and stearic. These fatty acids, similar to the mitochondrial hydrophobic component, bind Ca2+ with high affinity in comparison with unsaturated fatty acids, saturated fatty acids with shorter aliphatic chains, phospholipids, and other lipids. Ca2+-binding is inhibited by Mg2+ but not by K+. For palmitic acid, the Kd for Ca2+ was 5 microM at pH 8.5 and 15 microM at pH 7.5, with the Bmax of 0.48 +/- 0.08 mmol/g. This corresponds to one Ca2+ ion for eight palmitic acid molecules. The data of IR spectroscopy confirm that Ca2+ does not form ionic bonds with palmitic and stearic acids under hydrophobic conditions. It has been found that in the presence of Ca2+, palmitic and stearic acids, but not unsaturated FFA induce a nonspecific permeability in black-lipid membranes. Addition of Ca2+ in order to induce the permeability transition, increases the extractable amount of palmitic and stearic acids, the effect being prevented by a phospholipase A2 inhibitor. The possible involvement of palmitic and stearic acids in the mitochondrial nonspecific permeability is discussed.

Calcium-binding properties of the mitochondrial channel-forming hydrophobic component.

A hydrophobic, low-molecular weight component extracted from mitochondria forms a Ca2+-activated ion channel in black-lipid membranes (Mironova et al., 1997). At pH 8.3-8.5, the component has a high-affinity binding site for Ca2+ with a Kd of 8 x 10(-6) M, while at pH 7.5 this Kd was decreased to 9 x 10(-5) M. Bmax for the Ca2+-binding site did not change significantly with pH. In the range studied, 0.2 +/- 0.06 mmol Ca2+/g component were bound or one calcium ion to eight molecules of the component. The Ca2+ binding was strongly decreased by 50-100 mM Na+, but not by K+. Treatment of mitochondria with CaCl2 prior to ethanolic extraction resulted in a high level of Ca2+-binding capacity of the partially purified component. Cyclosporin A, a specific inhibitor of the mitochondrial permeability transition, when added to the mitochondrial suspension, decreased the Ca2+-binding activity of the purified extract severalfold. The calcium-binding capability of the partially purified component correlates with its calcium-channel activity. This indicates that the channel-forming component might be involved in the permeability transition that stimulates its formation.

Backbone Dynamics of Triple-helical Collagen-like Structure.

Some details of the backbone dynamics in the collagen-like triple helix is discussed and the role of backbone dynamics in functioning collagen proteins is illustrated. On a series of oligotripeptides synthetic analogs of collagen formation of high-frequency vibrational backbone dynamics and low-frequency nonlinear backbone dynamics upon stepwise elongation of peptide chain have been described using infrared spectroscopy and hydrogen-exchange method. In the fully completed triple helix the level of high-frequency backbone dynamics is regulated firstly by contact interactions of adjacent atoms and chemical bounded groups, while the level of low-frequency large-amplitude backbone dynamics depends mainly on cooperative interactions attributed by conjugation of interpeptide hydrogen bonds. In native collagens the nonlinear large-amplitude dynamics following by non-denaturational micro-unfolding of the triple-helical structure appears to be under the natural selection control delivering an optimal condition for formation, functioning and utilization of collagen fibrils.

[Effect of chain length on formation of a double-layer structure of Z-(Gly-Pro-Gly)n-OMe oligotripeptides]. / Obrazovanie struktury dvoinogo sloia v riadu oligotripeptidov Z-(Gly-Pro-Gly)n-OMe pri posledovatel'nom udlinenii molekuliarnoi tsepi.

Effect of chain length on the formation of the double-layered sheet in synthetic oligotripeptides Z-(Gly-Pro-Gly)n-OMe (n = 1,2,3,4) in the solid state has been studied by the IR-spectroscopy method. The major spectral features characteristic for the polymer structure was found in the oligomer spectra beginning with that of hexapeptide (n = 2). So a minimum peptide length when peptide chains are still able to adopt the double-layered sheet conformation in the solid state includes two triplets. The result is similar to that we have revealed early by investigation of the collagen-like triple helix formation in the oligomer series Z-(Gly-Pro-Pro)n-OMe. The double-layered sheet structure of the hexapeptide was found completely regular as that of the polymer. So the formation of the double-layered sheet structure unlike the formation of the triple-helical structure occurs at once without gradual ordering of interpeptide bonds and molecular packing by further increasing of the triplet number in oligomer chains. Additional information concerning specific incorporation of bound water in the double-layered sheet and water influence on peptide packing has been derived from analyses of hydration effect on the oligotripeptide IR-spectra especially on the components of the amide I band.

[Thermodynamic studies of triple-helical structures of the collagen type in oligotripeptides during study of molecular chain elongation]. / Termodinamicheskie issledovaniia trekhspiral'noi struktury kollagenovogo tipa v oligotripeptidakh pri posledovatel'nom udlinenii molekuliarnoi tsepi.

The conformational transition collagen-like triple helix in equilibrium with chains of oligotripeptides Z-(Gly-Pro-Pro)n-OMe with n = 6, 7, 8 in water by variation of solution temperature and sample concentration has been studied using IR-, CD-spectroscopy and microcalorimetry methods. The straight line correlation between the obtained value of the transition enthalpy and entropy and the number of the triplets (3n - 2), involved in the interpeptide set of hydrogen bonds was revealed. Evidently the effect of terminal groups is really weak in this case, and the interpeptide bonds of the triple helix may be regarded as equivalent one another. The estimated cooperative block of nucleation corresponds in length to the one full turn of the superhelix. The state diagrams of the oligotripeptides with n = 6, 7, 8 in aqueous solution are presented.

[Effect of the length of molecular chain on the formation of triple-helical collagen-like complex in solutions. The role of water in the formation of triple helix]. / Vliianie dliny molekuliarnoi tsepi na obrazovanie trekhspiral'nogo kompleksa kollagenovogo tipa v rastvorakh. Rol' vody v formirovanii troinoi spirali.

Effect of molecular chain length on the formation of the collagen-like triple helix in synthetic oligotripeptides Z-(Gly-Pro-Pro)n-OMe (n-1,2,3,...,8) in solutions has been studied, using IR- and CD-spectroscopy methods. The helix formation under different conditions is investigated: in the presence of a relatively inert solvent (chloroform), in the presence of a hydrogen bond acceptor (dioxan), in the presence of a hydrogen bond acceptor and donor as well (ethanol). Special attention is paid to the role of water in the formation of a stable triple-helical structure. Successive stages in the formation of a stable triple-helical structure in solutions during elongation of the peptide chain is revealed, which may be correlated with the helix nucleation process. A minimum peptide length, when peptide chains are still able to associate in the collagen-like triple-helical complex, involves three triplets for oligomers in chloroform solution, four triplets for oligomers in dioxan and ethanol solutions, six triplets for oligomers in aqueous solution. The main features of the completed triple-helical structure in water are found already in the oligotripeptide with n-8, where the formation one full turn of the superhelix is finished.

Bound water in the collagen-like triple-helical structure.

The ir amide bands of the triple-helical polytripeptides and collagens upon hydration of films are investigated. On the basis of our assignment of the amide I components, the formation of hydrogen bonds between the peptide backbone and structural water is studied. The C1O1--HOH hydrogen bonds are found more ordered than the C3O3--HOH hydrogen bonds. The specific incorporation of water in the triple helix is followed by multistep conformational changes and by increasing of the interpeptide hydrogen-bond strength. The formation of the polypeptide hydrate structure depending on the amino acid composition and the chain length is examined.