Antigenic Peptide-Thioredoxin Fusion Chimeras for In Vitro Stimulus of CD4+ TCR+ Jurkat T Cells.

Study of CD4+ T cell response and T cell receptor (TCR) specificity is crucial for understanding etiology of immune-mediated diseases and developing targeted therapies. However, solubility, accessibility, and stability of synthetic antigenic peptides used in T cell assays may be a critical point in such studies. Here we present a T cell activation reporter system using recombinant proteins containing antigenic epitopes fused with bacterial thioredoxin (trx-peptides) and obtained by bacterial expression. We report that co-incubation of CD4+ HA1.7 TCR+ reporter Jurkat 76 TRP cells with CD80+ HLA-DRB1*01:01+ HeLa cells or CD4+ Ob.1A12 TCR+ Jurkat 76 TRP with CD80+ HLA-DRB1*15:01+ HeLa cells resulted in activation of reporter Jurkat 76 TPR after addition of recombinant trx-peptide fusion proteins, containing TCR-specific epitopes. Trx-peptides were comparable with corresponding synthetic peptides in their capacity to activate Jurkat 76 TPR. These data demonstrate that thioredoxin as a carrier protein (trx) for antigenic peptides exhibits minimal interference with recognition of MHC-specific peptides by TCRs and consequent T cell activation. Our findings highlight potential feasibility of trx-peptides as a reagent for assessing the immunogenicity of antigenic fragments.

[The impact of masticatory muscles hypertonicity on the bite formation]. / Vliyanie gipertonusa zhevatel'noi muskulatury na formirovanie okklyuzii.

THE AIM OF THE STUDY: Was to assess the impact of masticatory muscles hypertonicity on the bite formation. MATERIALS AND METHODS: The study comprised 60 patients aged 7-14 years. Group 1 consisted of 20 individuals with Angle class 1 occlusion without masticatory muscle hypertonicity. Group 2 comprised 20 patients with class II malocclusion with hypertonicity of the masticatory muscles, group 3 - 20 patients class II malocclusion and no hypertonic masticatory muscles. All patients were examined according to common diagnostic protocol that included electromyography of the temporal and masticatory muscles at rest and in dynamics). RESULTS: In group 1 the mean IMPACT at rest was 242.8±133.6 µV, IMPACT during contraction was 880.50±201.5 µV; in group 2 - 797.9±413.0 and 1561.23±568.0 µV; in group 3 - 236.7±93.5 and 955.60±295.5 µV, correspondingly. The ratio of the activity of the temporal muscles to the masticatory muscles with neutral occlusion at rest correlates as 1:09, with compression 1:1. In patients with distal occlusion and the presence of hypertonicity at rest, the temporal muscles correspond to the chewing proper as 1:0.8, and with compression 1:09. CONCLUSION: The estimated ratio can contribute to the retroposition of the mandible, as well as inhibition of the growth of the mandible in the sagittal direction.

[Cephalometric CBCT analysis of the upper airways]. / Tsefalometricheskii analiz prosveta verkhnikh dykhatel'nykh putei s pomoshch'yu konusno-luchevoi komp'yuternoi tomografii.

The purpose of this literature review is to optimize the use of cephalometric diagnostic methods for assessing the condition of the upper respiratory tract. The article examines the areas of the upper respiratory tract associated with obstructive sleep apnea syndrome (OSA). Cephalometric analysis can be a useful tool for screening this pathology. However, it should be noted that there is still no clear understanding of the results of cephalometric changes in OSA, and therefore further research is necessary.

Morphological changes in salivary glands of neonatal rats after intra-abdominal hypertension.

OBJECTIVE: Given the overall prevalence of elevated Intra-abdominal pressure (IAP), along with earlier detection and appropriate therapy of Intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS), a significant reduction in patient morbidity and mortality is currently achieved by modern medicine. This article assesses the long-term degree of salivary gland damage in rats depending on the severity of experimental IAH during the neonatal period. MATERIALS AND METHODS: To simulate IAH, newborn rats, under the control of intravesical manometry, were injected into the abdominal cavity with bulking collagen filler in the amount necessary to create a given level of IAP. RESULTS: As shown by the results obtained, rats exposed to intra-abdominal hypertension for ten days had pathological changes in their salivary glands within 120 days. The severity of sialadenitis revealed a correlation with the severity of IAH. Some rats had individual reactions expressed in relative resistance to their organs' abnormalities under hypoxia. CONCLUSIONS: It was concluded that children with severe IAH history might need the disease prevention of the CNS, kidneys, digestive, and respiratory systems and oral diseases, particularly diseases involving the salivary glands. Future research is supposed to investigate further the IAH effect on various organs and tissues, including the dentofacial system.

Peculiarities of the Presentation of the Encephalitogenic MBP Peptide by HLA-DR Complexes Providing Protection and Predisposition to Multiple Sclerosis.

Predisposition to multiple sclerosis (MS), a chronic autoimmune disease of the central nervous system, is due to various factors. The genetic component is considered one of the most important factors. HLA class II genes contribute the most to the development of MS. The HLA-DRB1*15 allele group is considered one of the main genetic risk factors predisposing to MS. The group of HLA-DRB1*01 alleles was shown to have a protective effect against this disease in the Russian population. In this work, we compared the binding of the encephalitogenic fragment of the myelin basic protein (MBP) to two HLA-DR complexes that provide protection against and predisposition to MS: HLA-DR1 (HLA-DRB1*0101) and HLA-DR15 (HLA-DRB1*1501), respectively. We found that the myelin peptide MBP88-100 binds to HLA-DR1 at a rate almost an order of magnitude lower than the viral peptide of hemagglutinin (HA). The same was true for the binding of MBP85-97 to HLA-DR15 in comparison with viral pp65. The structure of the C-terminal part of the peptide plays a key role in the binding to HLA-DR1 for equally high-affinity N-terminal regions of the peptides. The IC50 of the myelin peptide MBP88-100 competing with viral HA for binding to HLA-DR1 is almost an order of magnitude higher than that of HA. As for HA, the same was also true for the binding of MBP85-97 to HLA-DR15 in comparison with viral pp65. Thus, autoantigenic MBP cannot compete with the viral peptide for binding to protective HLA-DR1. However, it is more competitive than viral peptide for HLA-DR15.

[Protocol of cone beam computed tomography analysis for patients with craniomandibular dysfunction]. / Protokol analiza konusno-luchevoi komp'yuternoi tomografii u bol'nykh s kraniomandibulyarnoi disfunktsiei.

The article presents a detailed protocol of cone beam computed tomography (CBCT), which includes not only the classic analysis of dentoalveolar and skeletal disorders, but diagnosis of cranial patterns (craniocervical cephalometric analysis by M. Rocabado, cephalometric diagnosis by Sassouni PLUS), analysis of transverse jaw sizes (Penn analysis) and measuring the volume of the airway for screening of obstructive sleep apnea. The literature review was conducted using the PubMed in the Scopus and Medline electronic databases through April 2020. Combining several methods of interpretation of CBCT in a single protocol will allow doctors to obtain a more complete amount of information to make a complete plan of comprehensive treatment.

[Use of fixed individual micro-implant supported orthodontic appliance in children with bilateral cleft lip and palate]. / Primenenie nes"emnykh individual'nykh ortodonticheskikh apparatov s oporoi na mikroimplantatakh u detei s dvustoronnei rasshchelinoi guby i neba.

Congenital malformations of the maxillofacial region are significant, not completely decisive, medical and social problems. Recent literature data indicate a trend towards improvement. PURPOSE OF THE STUDY: Improving the effectiveness of treatment of children with bilateral orthodontic and surgical training. MATERIAL AND METHODS: Under the supervision were 80 children with cleft lip and palate, with an age of up to 3 years. Of these 56 patients, 28 patients are the 2nd group prepared for surgery according to the developed technique. RESULTS: Preoperative orthodontic preparation of 28 children with bilateral cleft and an indicator that 22 (78.6%) patients should not have fully achieved results between the interhuman and fragmentary alveolar process of the upper jaw. In the second group of patients, 27 (96.4%) patients showed a normal ratio of the intermaxillary bone and lateral fragments. CONCLUSION: The use of the developed orthodontic design with active elements and mini implants in 96.4% of cases requires the presence of pre-spiral orthodontic preparation, normalization of the position of the intercellular bone and shape, followed by primary chelorinoplastics and in relation to additional uranoplastics, as well as the periodic stages of rehabilitation of patients with bilateral cleft lip and achieved thereby a stable aesthetic and functional result.

Biochar type and pyrolysis temperature effects on soil quality indicators and structural stability.

Quality of soils of the arid zones with low organic matter can be improved through the application of natural amendments especially biochar from various available feedstocks. The objective of this study was to evaluate the impacts of corn residue and poultry manure and their biochars on soil organic carbon (SOC), hot-water soluble carbohydrates (HWSC), basal soil respiration (BSR),and structural stability determined by HEMC (high-energy moisture characteristic). A sandy loam soil in pots were thoroughly mixed with 1, 2 and 4% w/w of corn residues (CR) and poultry manure (PM) feedstock and their biochars prepared at 350 and 650 °C of slow pyrolysis. Maize seeds were planted in pots and grown until physiological maturity when soil characteristics were measured. Treatments considerably altered the means of studied soil quality indicators, and increased SOC (1.5-10 times) and HWSC (1-7 times), and HEMC indices: volume of drainable pores ratio (VDPR, 1.5- 3.5 times), and stability ratio (SR, 1-3 times). Increasing pyrolysis temperature, regardless of the type and rate of feedstock, significantly decreased the SOC, BSR and percent of water-stable aggregates, and consequently structural stability indices. Contribution of both PM feedstock and its biochars was less effective than the CR ones (particularly the biochar produced at higher pyrolysis temperature), due to elevated sodium adsorption ratio (SAR) associated with higher slaking, physico-chemical dispersion and lower aggregate and structural stability.

An MDS-specific frailty index based on cumulative deficits adds independent prognostic information to clinical prognostic scoring.

The frailty index (FI) is based on the principle that the more deficits an individual has, the greater their risk of adverse outcomes. It is expressed as a ratio of the number of deficits present to the total number of deficits considered. We developed an MDS-specific FI using a prospective MDS registry and assessed its ability to add prognostic power to conventional prognostic scores in MDS. The 42 deficits included in this FI included measurements of physical performance, comorbidities, laboratory values, instrumental activities of daily living, quality of life and performance status. Of 644 patients, 440 were eligible for FI calculation. The median FI score was 0.25 (range 0.05-0.67), correlated with age and IPSS/IPSS-R risk scores and discriminated overall survival. With a follow-up of 20 months, survival was 27 months (95% CI 24-30.4). By multivariate analysis, age >70, FI, transfusion dependence, and IPSS were significant covariates associated with OS. The incremental discrimination improvement of the frailty index was 37%. We derived a prognostic score with five risk groups and distinct survivals ranging from 7.4 months to not yet reached. If externally validated, the MDS-FI could be used as a tool to refine the risk stratification of current clinical prognostication models.

Loading Rate of Exogenous and Autoantigenic Determinants on Major Histocompatibility Complex Class II Mediates Resistance to Multiple Sclerosis.

Genetic analysis of thousands of patients with multiple sclerosis (MS) and healthy Russian donors showed that the carriage of groups of HLA-DRB1*15 and HLA-DRB1*03 alleles is associated with the risk of MS, whereas the carriage of groups of HLA-DRB1*01 and HLA-DRB1*11 alleles is protective. Recombinant HLA-DRB1*01:01 with a high affinity can recognize the fragments of myelin basic protein (MBP), one of the autoantigens in MS. However, the comparison of the kinetic parameters of the load of MBP and viral HA peptides on HLA-DRB1*01:01, which is catalyzed by HLA-DM, showed a significantly lower rate of exchange of CLIP for MBP peptides. We assume that the observed protective properties of the group of HLA-DRB1*01 alleles may be directly associated with the ability of HLA-DRB1*01:01 to kinetically distinguish peptides of exogenous and endogenous nature.

Prostate-specific Antigen Bounce After Stereotactic Body Radiotherapy for Prostate Cancer: A Pooled Analysis of Four Prospective Trials.

AIMS: We conducted a pooled analysis of four prospective stereotactic body radiotherapy (SBRT) trials of low- and intermediate-risk prostate cancer to evaluate the incidence of prostate-specific antigen (PSA) bounce and its correlation with the time-dose-fraction schedule. The correlation between bounce with PSA response at 4 years (nadir PSA < 0.4 ng/ml) and biochemical failure-free survival (BFFS) was also explored. MATERIALS AND METHODS: The study included four treatment groups: 35 Gy/five fractions once per week (QW) (TG-1; n = 84); 40 Gy/five fractions QW (TG-2; n = 100); 40 Gy/five fractions every other day (TG-3; n = 73); and 26 Gy/two fractions QW (TG-4; n = 30). PSA bounce was defined as a rise in PSA by 0.2 ng/ml (nadir + 0.2) or 2 ng/ml (nadir + 2.0) above nadir followed by a decrease back to nadir. Patients with fewer than three follow-up PSA tests were excluded from the pooled analysis. RESULTS: In total, 287 patients were included, with a median follow-up of 5.0 years. The pooled 5-year cumulative incidence of bounce by nadir + 2.0 was 8%. The 2-year cumulative incidences of PSA bounce by nadir + 0.2 were 28.9, 21, 19.6 and 16.7% (P = 0.12) and by nadir + 2.0 were 7.2, 8, 2.7 and 6.7% (P = 0.32) for TG-1 to TG-4, respectively. Multivariable analysis revealed that for nadir + 2.0, pre-treatment PSA (odds ratio 0.49; 95% confidence interval 0.26-0.97) correlated with PSA bounce. Although PSA bounce by nadir + 0.2 (odds ratio 0.10; 95% confidence interval 0.04-0.24) and nadir + 2.0 (odds ratio 0.29; 95% confidence interval 0.09-0.93) was associated with a lower probability of PSA response at 4 years, there was no association between bounce by nadir + 0.2 (hazard ratio 0.36; 95% confidence interval 0.08-1.74) or nadir + 2 (hazard ratio 1.77; 95% confidence interval 0.28-11.07) with BFFS. CONCLUSION: The incidence of PSA bounce was independent of time-dose-fraction schedule for prostate SBRT. One in 13 patients experienced a bounce high enough to be misinterpreted as biochemical failure, and clinicians should avoid early salvage interventions in these patients. There was no association between PSA bounce and BFFS.

The Contribution of Major Histocompatibility Complex Class II Genes to an Association with Autoimmune Diseases.

Genetic studies of patients with autoimmune diseases have shown that one of the most important roles in the developing of these diseases is played by a cluster of genes of the major histocompatibility complex (MHC), as compared with other genome areas. Information on the specific contribution of MHC alleles, mostly MHC class II ones, to the genetic predisposition to autoimmune diseases is crucial for understanding their pathogenesis. This review dwells on the most relevant aspects of this problem: namely, the correlation between carriage of certain MHC II alleles and an increased (positively associated allele) or reduced (negatively associated allele) probability of developing the most common autoimmune diseases, such as type 1 diabetes, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, autoimmune thyroiditis, etc. The most universal haplotypes, DR3-DQ2 and DR4-DQ8, are positively associated with many of these diseases, while the universal allele HLA-DRB1*0701 is protective.

[Clinical and diagnostic rationale for the final stage of orthodontic treatment]. / Kliniko-diagnosticheskoe obosnovanie zavershaiushchego étapa ortodonticheskogo lecheniia.

The aim of the study was to improve the efficiency of the final stage of orthodontic treatment. The study included 32 patients divided into two groups of 16. Orthodontic treatment was performed using a bracket system active self-ligation straight arc technique with step adjustment anthropometric quantitative evaluation system (The ABO Model Grading System). Centric occlusion and centric relation were analyzed at baseline and after treatment by mandible position indicator MPI (Mandibular Position Indicator). The first group received treatment according to conventional protocol, in the second group braces were readjusted by indirect bonding after ABO anthropometric measurements. The efficiency of orthodontic treatment using ABO system at the late stages of adjustment in the second group was significantly higher than in the first group (p=0.002248; p=0.023431, respectively). MPI analysis in the first group in 10 (62.5%) patients showed mismatch of centric occlusion and centric relation not seen in the second group. In 3 (18.75%) patients of group 2 the difference did not exceed 0.5 mm which was considered normal. Thus, the application of ABO anthropometric system followed by readjustment of incorrectly positioned brackets by indirect bonding promotes MPI values improvement, which in turn allows getting the most accurate, and stable results of orthodontic treatment.

Long-term Toxicity and Health-related Quality of Life after Single-fraction High Dose Rate Brachytherapy Boost and Hypofractionated External Beam Radiotherapy for Intermediate-risk Prostate Cancer.

AIMS: To report health-related quality of life (HRQOL) and toxicity in prostate cancer patients treated with single-fraction high dose rate (HDR) brachytherapy boost and external beam radiotherapy (EBRT). MATERIALS AND METHODS: Patients with intermediate-risk prostate cancer were accrued to a phase II clinical trial of 15 Gy HDR boost and EBRT to a dose of 37.5 Gy in 15 fractions. HRQOL (Expanded Prostate Cancer Index Composite [EPIC]), urinary symptoms (International Prostate Symptom Score [IPSS]), erectile function (International Index of Erectile Function [IIEF]) and toxicity (Common Terminology Criteria for Adverse Events [CTCAE], version 3.0) were monitored prospectively. Univariate and multivariate logistic regression analysis was used to investigate associations between HRQOL/toxicity and baseline covariates. RESULTS: The median follow-up time was 5.2 years. The change in the median EPIC scores from baseline to year 5 in the urinary domain was from 91 to 85 (P = 0.0028), in the bowel domain was from 98 to 96 (P = 0.03), in the sexual domain was from 63 to 35 (P < 0.0001) and the hormonal domain remained unchanged at 95 (P = 0.93). Fifty-nine per cent and 46% of the patients with normal erectile function at baseline remained potent at year 1 and year 5, respectively. Late genitourinary toxicity grade 1, 2 and ≥3 occurred in 29, 59 and 4% of patients, respectively. The rates of late gastrointestinal toxicity grade 1, 2 and ≥3 were documented as 45, 19 and 0%, respectively. On multivariate logistic regression analysis, patients with larger prostates were more likely to develop a urinary late toxicity grade ≥2 (P = 0.01). The dose to 10% of the urethra was the only factor associated with a decline in the EPIC urinary domain score (P = 0.012). Prostate volume >43 ml was associated with higher late genitourinary toxicity grade ≥2. CONCLUSIONS: Single 15 Gy HDR brachytherapy with EBRT has a low rate of late genitourinary and gastrointestinal toxicities. Late urinary morbidity may be minimised by limiting the dose to the urethra, particularly for patients with larger prostates.

Prophylactic RhCE and Kell antigen matching: impact on alloimmunization in transfusion-dependent patients with myelodysplastic syndromes.

BACKGROUND AND OBJECTIVES: Thirty to 80 per cent of patients with myelodysplastic syndromes (MDS) become transfusion-dependent and are at risk for red blood cell (RBC) alloimmunization. This study compared alloimmunization rates in transfusion-dependent patients with MDS at an institution with a policy of prophylactic antigen matching for RhCE and K (PAM) with those transfused at institutions without such a policy (non-PAM). MATERIALS AND METHODS: Transfusion records were retrospectively reviewed to determine total number of RBC transfusions received, whether RBC phenotyping was performed, the type and date of first alloantibody development and receipt of prophylactic antigen matching for RhCE and K. RESULTS: In 176 transfusion-dependent patients with MDS, the overall rate of new alloimmunization was 17%; the majority of patients (87%) developed at least one alloantibody to Rh or Kell antigens. The alloimmunization rate at the institution with a PAM policy was 11% compared with 23% at non-PAM institutions (P = 0·06). The rate of Rh/K alloimmunization was 7 vs. 22%, respectively (P = 0·008). No patient who received PAM developed a Rh/K alloantibody. CONCLUSION: The rate of alloimmunization was 11% at an institution with a PAM policy which was non-significantly lower than 23% at institutions without a PAM policy. However, rates of Rh/K alloimmunization were significantly lower. Such a policy should be considered in transfusion-dependent patients with MDS, although further studies on cost-effectiveness and careful consideration of resource availability in the local context are required.

Heterodimer HLA-DM Fused with Constant Fragment of the Heavy Chain of the Human Immunoglobulin Accelerates Influenza Hemagglutinin HA306-318 Loading to HLA-DR1.

Major histocompatibility complex class II (MHC II) plays an important role not only in the adaptive immune responses to foreign pathogens, but also in the development of some autoimmune diseases. Non-classical MHC, HLA-DM is directly involved in MHC II loading with the peptide. To study this process, we synthesized recombinant proteins HLA-DR1 and HLA-DM. α/ß-Chains of DR1 heterodimer contained C-terminal leucine domains of the fos and jun factors, respectively. Each DM chain contained constant fragment of human antibody heavy chain fused via a long linker domain. In addition, DM α-chain carried N165D substitution suppressing potential glycosylation at this site. We observed significant acceleration of DR1 peptide loading with influenza HA306-318 hemagglutinin in the presence of DM, which indicates functionality of recombinant DR1-DM protein couple. Our results can be used to study the presentation of other viral and self-antigens and can become the basis for the development of new drug modeling.

Accelerated hypofractioned postoperative radiotherapy for prostate cancer: a prospective phase I/II study.

AIMS: To present the initial findings of a single institution, phase I/II study investigating hypofractionated radiotherapy in patients undergoing post-prostatectomy treatment. MATERIALS AND METHODS: Patients requiring postoperative radiotherapy were prospectively enrolled. Dose was prescribed to the prostate bed with 51 Gy in 17 daily fractions. Androgen deprivation was optional. Acute and late gastrointestinal/genitourinary toxicity were assessed using the National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0 and quality of life was assessed using the Expanded Prostate Cancer Index Composite evaluation tool. Prostate-specific antigen (PSA) was evaluated at every follow-up. RESULTS: Thirty patients were enrolled between 2009 and 2011. The median age was 65 years and most had Gleason 7 disease (86%) with pT2c or pT3a (82%). Positive margins were documented in 67% of the patients. The median pre-treatment PSA was 0.12 ng/ml. The median follow-up was 24 months. Overall toxicity was low, with >80% of patients having ≤ grade 1 acute toxicity in both genitourinary and gastrointestinal realms. Similarly, only two patients (6%) experienced grade 2/3 late gastrointestinal/genitourinary toxicity. Quality of life scores were also indicative of a well-tolerated treatment. PSA failure was seen in five patients (17%). CONCLUSIONS: We present a hypofractionated schedule of postoperative prostate radiotherapy that is both well tolerated in terms of both toxicity and quality of life measures. Initial PSA control is encouraging. Further evaluation with a longer follow-up and a larger cohort is warranted.

[The experience of complex treatment of children with bilateral cleft lip and palate].

Three grades of premaxilla deformation were revealed in children with bilateral cleft lip and palate. All patients with grade I and some children with grade II deformation received early orthopedic treatment by Sharova appliance while in patients with grade III deformation orthodontic devices with bone fixation with microimplants were used or microimplants alone for orthodontic anchorage. This approach allowed achieving full side contact of premaxilla and maxillary fragments in 1-2 months thus creating favorable conditions for surgical procedure.