Serotype Distribution and Multi Locus Sequence Type (MLST) of Erythromycin-Resistant Streptococcus Pneumoniae Isolates in Tehran, Iran.

Background: The number of erythromycin-resistant Streptococcus pneumoniae has significantly increased around the world. The present study aimed to determine the serotype distribution and molecular epidemiology of the erythromycin-resistant Streptococcus pneumoniae (ERSP) isolated from patients with invasive disease. Methods: A total of 44 Streptococcus pneumoniae isolates were tested for susceptibility to several antimicrobial agents. Additionally, the polymerase chain reaction (PCR) was applied to evaluate ERSP isolates in terms of the presence of erythromycin resistance genes (e.g., ermB and mefA). The isolates were serotyped using the sequential multiplex-PCR method, and molecular epidemiology was assessed through the multilocus sequence typing (MLST) analysis. Results: The results represented multidrug resistance (MDR) in approximately half of the pneumococcal isolates. Among 22 ERSP isolates, 20 (90.9%) and 12 (56%) ones contained ermB and mefA, respectively. Further, 14 (31.8%), 3 (22.7%), and 19A (18.1%) were the common serotypes among the isolates. No significant correlation was observed between serotypes and erythromycin resistance genes. Furthermore, the MLST results revealed 18 different sequence types (STs), the top ones of which were ST3130 (3 isolates) and ST166 (3 isolates). Population genetic analysis disclosed that CC63 (32%), CC156 (18%), and CC320 (18%) were identified as the predominant clonal complexes. Conclusions: The ERSP isolates exhibited high genetic diversity. The large frequency of MDR isolates suggests the emergence of high resistant strains, as well as the need to implement vaccination in the immunization schedule of Iran. These accumulating evidences indicate that 13-valent pneumococcal conjugate vaccines provided higher serotype coverage in the ERSP isolates.

Objective measurement of Inferior-Directed stiffness in glenohumeral joint using a specially designed robotic device in healthy shoulders; Within- and Between-Session reliability.

Clinical assessment of capsuloligamentous structures of the glenohumeral joint has been qualitative and subjective in nature, as demonstrated by limited intra- and inter-rater reliability. Robotic devices were utilized to develop a clinically objective measurement technique for glenohumeral joint stiffness. The purpose of this study was to quantify the amount of inferior-direction stiffness of the glenohumeral joint using a safe clinical device in the asymptomatic individuals, and to determine between trial and between session reliability of the robotic device. Twenty healthy subjects were recruited via convenience sampling. Inferior-directed translation and applying force were measured using displacement and force sensors of a robotic device. The stiffness values were calculated as the mean of the slopes of the linear portions of the force-displacement curves for the cycles obtained after familiarization and preconditioning. Four trials for each measurement occasion were averaged to determine the stiffness value for each subject in one session. Repeatability of glenohumeral joint stiffness measurements for between trials and between two sessions was determined using intraclass correlation values and standard error of the measurements. The mean stiffness value was 1.50 N/mm (±0.40) and 1.52 N/mm (±0.40), respectively. The robotic device for stiffness assessment was reliable for repeated measures of stiffness in one session, and between sessions with ICC equal 0.96 (95% CI 0.93-0.98), and 0.97 (95% CI 0.95-0.99), respectively. The SEM between the trials was in each session 0.08 N/mm. The results of this study provide that our robotic technique for quantifying glenohumeral joint stiffness is precise and reproducible.

Impact of Laboratory Methods and Gene Targets on Detection of Streptococcus pneumoniae in Isolates and Clinical Specimens.

BACKGROUND: Timely identification of Streptococcus pneumoniae infections can lead to a decrease in mortality rates. Differentiation of S. pneumoniae from other similar species using traditional culture-based and molecular methods is problematic. In this study, we assessed the efficacy of identifying the blpA and lytA for the detection of S. pneumoniae from isolates and various clinical samples using molecular methods. METHODS: A total of 440 clinical samples were collected from patients with suspected invasive pneumococcal infections during February 2016 to October 2018. Biochemical tests were used to confirm the dubious colonies on 5% sheep blood agar. Fifty-seven confirmed isolates, 57 culture-positive samples, and 57 culture-negative samples were analyzed for the presence of blpA and lytA using both conventional and real-time PCR. RESULTS: All the isolates and culture-positive samples were positive for blpA and lytA by both PCR methods. Of the 57 culture-negative samples, conventional and real-time PCR amplified blpA from six and two samples, and lytA from seven and two samples, respectively. CONCLUSION: The specificity of real-time PCR assay was significantly higher than that of conventional PCR for the identification of S. pneumoniae. In addition, it is suggested that respiratory secretions are not suitable specimen for direct diagnosis of pneumococcal infections.

Autolysin (lytA) recombinant protein: a potential target for developing vaccines against pneumococcal infections.

PURPOSE: N-acetylmuramoyl-l-alanine amidase known as lytA, is an immunogenic protein that plays an important role in the pathogenesis of Streptococcus pneumoniae. It is highly conserved among S. pneumoniae strains and is absent among other Streptococcus species. In the present study, the level of antibodies against the lytA recombinant protein was evaluated in healthy individuals' sera. MATERIALS AND METHODS: DNA was extracted from S. pneumoniae ATCC 49619 to amplify lytA gene by polymerase chain reaction assay. The lytA amplicon and pET28a vector were separately double digested using Nde-1 and Xho1 restriction enzymes and then ligated together with ligase enzyme. The recombinant plasmid was expressed in Escherichia coli BL21 strain and the lytA recombinant protein purified using nickel-nitrilotriacetic acid affinity chromatography. Western blot was carried to detect lytA recombinant protein. Sixty healthy individual's sera (at three age groups: group 1, <2; group 2, 2-40; and group 3, 60-90 years old) were collected and the titers of anti-lytA antibodies were determined. RESULTS: The lytA gene was highly expressed in E. coli BL21 host. The recombinant lytA protein was purified and confirmed by western blotting. Tukey test analysis showed that there were no significant differences among the age groups considering the anti-lytA titer of 10. However, at the anti-lytA titer of 60, significant differences were observed between group 1 vs. group 2 (p<0.001); group 1 vs. group 3 (p=0.003), and group 2 vs. group 3 (p=0.024). CONCLUSION: The lytA protein seems to be a highly immunogenic antigen and a potential target for developing vaccines against pneumococcal infections.

Serotype Distribution and Antibiotic Susceptibility of Streptococcus pneumoniae Isolates in Tehran, Iran: A Surveillance Study.

INTRODUCTION: Encapsulated Streptococcus pneumoniae strains cause high morbidity and mortality, mainly in countries with no pneumococcal conjugate vaccines (PCVs) immunization program. This study investigated the epidemiological changes of S. pneumoniae isolates including serotype distribution and antimicrobial susceptibility in Tehran, Iran. METHODS: A total of 80 S. pneumoniae samples were collected from patients admitted to Shariati hospital over two periods. Half of the isolates were collected from February to September 2017 and the other half from July 2018 to March 2019. The antimicrobial susceptibility testing and PCV-13 serotype coverage of S. pneumoniae isolates were evaluated among patients with invasive and non-invasive infections. RESULTS: The most common serotypes were 23F (17.5%), 14 (16.3%), 3 (16.3%) 19F (12.5%), and 19A (12.5%) in the present study. The vaccine coverage rates of PCV-7, PCV-10 and PCV-13 were 52.6%, 52.6%, and 83.7%, respectively. S. pneumoniae isolates with the serotype of the PCV-13 showed an increasing trend during the study. Nearly half of the S. pneumoniae strains were MDR, while MDR serotype 19A increased (40%) during the study periods. A small minority of isolates (16%) belonged to non-vaccine serotypes, 65% of which were assigned to MDR. In general, the frequency of penicillin resistant and MDR strains were estimated about 27.5% and 51%, respectively. An increase was observed in resistance to erythromycin and co-trimoxazole. CONCLUSION: The results showed that majority of the circulating serotypes in our study are related to PCV-13 serotypes. The use of conjugate vaccine in the immunization program and surveillance of antimicrobial resistance can be effective in reducing the pneumococcal clinical burden.

Design and Preliminary Evaluation of a Novel Robotic System for Mobilization of Glenohumeral Joint.

Joints mobilization is an essential but subjective treatment in the physical therapy of the patients with joint hypomobility such as frozen shoulder. Recently many instrumented force and displacement indentations have been proposed for assessing and diagnosis of joints stiffness. The devices are not, however, feasible and applicable for use in clinical and therapeutic conditions considering the requirements of the joints mobilization principles in physiotherapy. This paper describes a novel design of a robotic system for mobilization of glenohumeral joint and the preliminary evaluation of mobilization robot in a subject with hypomobile glenohumeral joint. A new mechanism is presented which enables the robotic system to execute the mobilization maneuver in 1 inch linear motion path when it grasps the glenohumeral joint and holds the upper limb situated in the 90 degrees relaxed abduction. It was shown that the mobilization robot can be used effectively and practical for mobilization treatment. Furthermore such a device may be used as a diagnostic and assessing device for evaluating the stage of hypomobility based on Maitland method.

Fibrinogen and Fibronectin Binding Activity and Immunogenic Nature of Choline Binding Protein M.

BACKGROUND: Choline-binding proteins (CBPs) are a group of surface-exposed proteins, which play crucial and physiological roles in Streptococcus pneumoniae. The novel member of CBPs, choline-binding protein M (CbpM) may have binding activity to plasma proteins. This study aimed to clone and express CbpM and demonstrate its interaction with plasma proteins and patients' sera. METHODS: The total length of cbpM gene was cloned in pET21a vector and expressed in BL21 expression host. Verification of recombinant protein was evaluated by Western blot using anti-His tag monoclonal antibody. Binding ability of the recombinant protein to plasma proteins and the interaction with patients' sera were assessed by Western blot and ELISA methods. RESULTS: The cbpM gene was successfully cloned into pET21a and expressed in BL21 host. Binding activity to fibronectin and fibrinogen and antibody reaction of CbpM to patients' sera was demonstrated by Western blot and ELISA methods, respectively. CONCLUSION: CbpM is one of the pneumococcal surface-exposed proteins, which mediates pneumococcal binding to fibronectin and fibrinogen proteins.

Inter- and intrarater reliability of modified lateral scapular slide test in healthy athletic men.

OBJECTIVE: The reliability of lateral scapular slide test (LSST) at 90 degrees of abduction is controversial; therefore, in order to achieve more reliability it may be necessary to make changes in this particular position. METHODS: Modified lateral scapular slide test (MLSST) was done on thirty male basketball players with two examiners in one session and for the retest with one examiner in the next week. The test was done in 7 positions: arm relaxed at the side (P1), 90 degrees of abduction (P2), 90 degrees of scaption without having a weight in hands (P3), 90 degrees of scaption with having 3 different weights (1, 2, and 4 kg) in hands (P4, P5, and P6, resp.), and 180 degrees of scaption without having a weight in hands (P7). RESULTS: In P1 and P6, the ICC scores indicated the highest level of intrarater reliability. In P2, the ICC scores showed a fair level of intrarater reliability, as the minimum reliability. The maximum and minimum interrater reliability were P1 and P4, respectively. CONCLUSION: Scaption with loading, as a functional position in the overhead athletes, is a reliable positioning and may be replaced with the third position of the traditional LSST.

The Modified Tardieu Scale for the measurement of elbow flexor spasticity in adult patients with hemiplegia.

PRIMARY OBJECTIVE: To investigate the inter-rater reliability of the Modified Tardieu Scale (MTS) in the measurement of elbow flexor muscle spasticity in patients with adult hemiplegia. RESEARCH DESIGN: A cross-sectional study to examine the agreement between two raters on elbow flexor muscle spasticity using Modified Tardieu Scale. INTERVENTIONS: Not applicable. METHODS AND PROCEDURES: Thirty patients (21 men, nine women) with an age range of 21-79 years (mean = 54.9; SD = 15.7) were tested. Two inexperienced raters assessed the elbow flexor muscle spasticity in the affected side during the same session. The order of assessments by the two raters was randomized. The main outcome measure was dynamic component of spasticity (R2-R1). Reliability was also calculated for MTS scores, angle of muscle reaction (R1), and passive range of motion (R2). MAIN OUTCOMES AND RESULTS: The ICC for R2-R1 was 0.72. The ICC for MTS scores, R1 and R2 were 0.74, 0.74 and 0.56, respectively. CONCLUSIONS: The Modified Tardieu Scale did not provide acceptable high inter-rater reliability in the measurement of muscle spasticity in patients with hemiplegia when used by raters of limited experience.