Comparative evaluation of cervical pulpotomy and pulpectomy for primary molars with irreversible pulpitis: a multicentre randomised controlled trial.

PURPOSE: The objective of this multicentre randomised controlled trial was to compare the clinical/radiographic outcomes of cervical pulpotomy using calcium-enriched mixture cement (PCEM) and pulpectomy using Metapex (PM) in primary molars with irreversible pulpitis (IP). METHODS: A total of 134 primary molars from 94 children were randomly assigned to two intervention groups: the PCEM group (n = 74) and the PM group (n = 60). Baseline characteristics including age/gender/molar type/tooth type/jaw were recorded. The primary outcome measures were clinical/radiographic success rates assessed at the first and second follow-up appointments. Secondary outcomes included reasons for clinical/radiographic failures. Multiple logistic regression analysis was performed to determine the impact of various factors on the success rates. RESULTS: The mean age of the participants in both groups was similar (PCEM group: 5.4 years, PM group: 5.5 years). Gender distribution, molar type, tooth type, jaw, and number of practitioners were comparable between the groups. The clinical success rate at the first follow-up was 98.6% in the PCEM group and 96.4% in the PM group. At the second follow-up, the clinical success rate was 97.1% in the PCEM group and 91.1% in the PM group. The radiographic success rates at the first and second follow-up were 98.6% and 96.4% in the PCEM group and 96.4% and 91.1% in the PM group, respectively. Multiple logistic regression analysis did not reveal any significant association between the success rates and age/gender/molar type/jaw, or treatment groups (P > 0.05). CONCLUSION: In primary molars with IP, both simple/conservative cervical pulpotomy using calcium-enriched mixture cement and pulpectomy using Metapex demonstrated high clinical/radiographic success rates. No significant differences were observed between the two treatment modalities. These findings suggest that both techniques can be considered effective treatment options for managing primary molars with IP. TRIAL REGISTRATION NUMBER: Trial registration number: IRCT20201226049838N1, retrospectively registered on 12 January 2021.

Establishment of reverse transcription polymerase chain reaction assay for simultaneous detection of human parainfluenza virus types 1 to 4 in children with influenza like illnesses.

OBJECTIVES: The aim of this study was to develop an in-house multiplex reverse transcription polymerase chain reaction (mRT-PCR), which can recognize HPIV1-4 in clinical samples. BACKGROUND: Human parainfluenza virus (HPIV) is one of the major causes of viral respiratory infections and can affect people at any age, especially infants and young children. METHODS: Four sets of specific primers targeting conserved areas of hemagglutinin-neuraminidase (HN) genes of HPIV1-4, were designed and tested with type-related plasmid controls. Specificity and sensitivity of mPCR were tested. One-step mRT-PCR was set up using a viral panel containing 10 respiratory viruses, including HPIVs. One hundred nasopharyngeal samples of respiratory infection patients were tested using the set One-step mRT-PCR. RESULTS: The specificity of set mPCR for HPIV1-4 using plasmid positive controls was proved and reaction sensitivity was measured. The specificity of set mRT-PCR was confirmed and 4 and 5 out of 100 clinical samples were HPIV1 and HPIV2 positive, respectively. CONCLUSION: The developed one-step mRT-PCR in this study is an effective and specific assay for clinical diagnosis of HPIV1 to 4 (Tab. 1, Fig. 6, Ref. 28).

Generation of GFP Native Protein for Detection of Its Intracellular Uptake by Cell-Penetrating Peptides.

Different types of lipid- and polymer-based vectors have been developed to deliver proteins into cells, but these methods showed relatively poor efficiency. Recently, a group of short, highly basic peptides known as cell-penetrating peptides (CPPs) were used to carry polypeptides and proteins into cells. In this study, expression and purification of GFP protein was performed using the prokaryotic pET expression system. We used two amphipathic CPPs (Pep-1 and CADY-2) as a novel delivery system to transfer the GFP protein into cells. The morphological features of the CPP/GFP complexes were studied by scanning electron microscopy (SEM), Zetasizer, and SDS-PAGE. The efficiency of GFP transfection using Pep-1 and CADY-2 peptides and TurboFect reagent was compared with FITC-antibody protein control delivered by these transfection vehicles in the HEK-293T cell line. SEM data confirmed formation of discrete nanoparticles with a diameter of below 300 nm. Moreover, formation of the complexes was detected using SDS-PAGE as two individual bands, indicating non-covalent interaction. The size and homogeneity of Pep-1/GFP and CADY-2/GFP complexes were dependent on the ratio of peptide/cargo formulations, and responsible for their biological efficiency. The cells transfected by Pep-1/GFP and CADY-2/GFP complexes at a molar ratio of 20 : 1 demonstrated spreading green regions using fluorescent microscopy. Flow cytometry results showed that the transfection efficiency of Pep-based nanoparticles was similar to CADY-based nanoparticles and comparable with TurboFect-protein complexes. These data open an efficient way for future therapeutic purposes.

Expression of HA1 antigen of H5N1 influenza virus as a potent candidate for vaccine in bacterial system.

The impending influenza virus pandemic requires global vaccination to prevent large-scale mortality and morbidity, but traditional influenza virus vaccine production is too slow for rapid responses. In this study, bacterial system has been developed for expression and purification of properly folded HA1 antigen as a rapid response to emerging pandemic strains. Here, a recombinant H5N1 (A/Indonesia/05/05) hemagglutinin globular domain, the synthesized HA1 (1-320 amino acids), was amplified and cloned into pET-28a bacterial expression vector. Then, his-tagged HA1 protein was expressed in Escherichia coli BL21 under 1 mM IPTG induction. The protein expression was optimized under a time-course induction study and further purified using Ni-NTA chromatography. Migration size of protein was detected at 40 KDa by Western blot using anti-His tag monoclonal antibody and demonstrated no discrepancy compared to its calculated molecular weight. Since most antigenic sites are in the HA1 domain of HA, using this domain of influenza virus as antigen is of great importance in vaccine development. The ability of the antibody stimulation against HA1 expressed in bacterial cells is also examined using enzyme-linked immunosorbent assay (ELISA) analysis. Upon immunization of rabbits, oligomeric HA1 elicited potent neutralizing antibodies and high levels of serum antibody binding to HA1. Our findings suggest that HA1-based vaccines can be produced efficiently in bacterial systems and can be easily upscaled in response to a pandemic influenza virus threat.

Expression of antigenic determinants of the haemagglutinin large subunit of novel influenza virus in insect cells.

The global outbreak of novel A/H1N1 spread in human population worldwide has revealed an emergency need for producing a vaccine against this virus. Current influenza vaccines encounter problems with safety issues and weak response in high-risk population. It has been established that haemagglutinin is the most important viral antigen to which antibody responses are directed, and recombinant subunit vaccines, haemagglutinin of influenza A and B viruses, have been considered in order to facilitate vaccine production. In the present study, we have focused on construction of a recombinant baculovirus encoding the large subunit of novel influenza virus A/H1N1 haemagglutinin. The full genome of haemagglutinin was cloned into pGEM-TEasy vector and sequenced. The large subunit of the haemagglutinin gene was amplified by PCR using specific primers and cloned into pFast- BacHTc donor plasmid, which was then confirmed by restriction enzyme analysis and sequencing and transformed into E. coli DH10Bac competent cells. The bacmid DNA was transfected into insect cells to produce recombinant baculovirus. Expression of recombinant haemagglutinin in insect cells was determined by SDS-PAGE and immunoblotting. It has been shown that the recombinant haemagglutinin (rHA) obtained from the baculovirus insect cell expression system has suitable immunogenicity in human and can be considered as a candidate flu vac- cine. Here we produced large amounts of the HA1 protein of novel influenza A/H1N1 (Iranian isolate) in insect cells. The immunogenicity and efficacy of the recombinant HA1 will be evaluated as a vaccine candidate and compared to the recombinant HA1 produced in a prokaryotic system.

The role of 99mTc-MIBI scintigraphy in the management of patients with pulmonary tuberculosis.

BACKGROUND: This study aimed to determine whether 99mTc-methoxyisobutylisonitrile (MIBI) scanning could improve diagnostic accuracy of pulmonary tuberculosis (PTB) and help clinical decision making for an accurate management. MATERIAL AND METHODS: 99mTc-MIBI scintigraphy was performed in 62 cases of PTB 34 cases had active pulmonary tuberculosis (APTB) and were at the beginning of antituberculosis medication (group 1) as well as 28 cases had inactive pulmonary tuberculosis (IPTB) and were post antituberculosis medication (group 2). The qualitative and semiquantitative findings of both scanning methods were assessed. For semiquantitative evaluation, regions of interest (ROIs) were drawn over the lesion (L), non-lesion (NL) and neck soft tissue (NST). The mean count values of ROIs were obtained and L/NL and L/NST were calculated. RESULTS: Thirty-four patients with APTB (15 males and 19 females; mean age of 47.85 +/- 1.91 yrs) and 28 cases with IPTB (9 male and 19 females; mean age of 53.96 +/- 2.33 yrs) were included in this study. The sensitivity, specificity, accuracy, positive and negative predictive (PPV and NPV) values of 99mTc-MIBI were 88.2%, 75%, 82.2%, 81.1% and 84% respectively. The mean value of L/NL in the APTB for 99mTc-MIBI was 1.45 +/- 0.18 and L/NST was 1.57 +/- 0.26 which was significant statistically (p < 0.00). CONCLUSIONS: The study demonstrated that 99mTc-MIBI scanning can be complementary to other diagnostic techniques especially in patients with indeterminate APTB and those in whom recurrent disease is suspected. In addition, because of its availability, rather low costs, easy performance, and objective semiquantitative information supplied, 99mTc-MIBI scanning might be establish in routine imaging center to assess the pulmonary tuberculosis. However, further exploration is needed to validate its clinical role.

Prevalence of HI antibody titer against rubella virus to determine the effect of mass vaccination in Tehran.

BACKGROUND: Rubella is an infectious viral disease, has a worldwide distribution and is normally a mild childhood disease. Infection during early pregnancy may cause fetal death or congenital rubella syndrome. The highest risk of CRS is found in countries with high susceptibility rates among women of childbearing age. In many developed and some developing countries, large-scale rubella vaccination during the past decade has drastically reduced or practically eliminated rubella and CRS. Mass vaccination campaigns and Expanded Program of Immunization (EPI) have increased vaccine coverage in the world with a substantial impact on the reduction of rubella infections, such as CRS. OBJECTIVE: The present study was preformed to evaluate the immune status against rubella before and after the mass campaign vaccination on 22 December 2003. STUDY DESIGN: A total of 320 samples were collected from the healthy subjects before and after the vaccination and 80 paired sera were collected and tested for the presence of rubella antibody using HI test. RESULTS AND CONCLUSIONS: Based on the results, 98.1% of the population has gained anti-rubella antibody, compared with 92.2% before the vaccination. The data revealed that 98.75% of the paired subjects had rubella antibody after mass vaccination which is statistically significant.

Estimation of right ventricular pressure by 201Tl scintigraphy in paediatric cardiac disease.

To estimate right ventricular overload in paediatric cardiac disease, 201Tl scintigraphy was used to quantify right ventricular uptake. Six methods for calculating the 201Tl right-to-left ventricular uptake ratio (TRL) were compared, based on the location of regions of interest (ROIs), use of total count or mean count density, and the ROI-based method or profile method. When the TRL was compared with the right-to-left ventricular peak systolic pressure ratio (RVP/LVP), the mean count density method using entire right and left ventricular ROIs seemed to be appropriate, considering its simplicity, reproducibility and regression line. The linear regression line was RVP/LVP = 1.15 x TRL - 0.13 (n = 15, P = -0.0001), and RVP(mm Hg) = 162 x TRL - 33 (n = 15, P = 0.0001). In four patients with pulmonary stenosis, the changes of TRL were in good agreement with the right ventricular pressure changes after percutaneous balloon valvuloplasty. Thus, evaluation of thallium scintigraphy using this quantitative method can be a simple and effective way to evaluate patients with right ventricular pressure overloading.