Antibodies Produced Toward Recombinant RBD and Nucleocapsid Neutralize SARS-COV-2.

Background: The highly contagious SARS-COV-2 virus spread rapidly from China and formed a global pandemic. The virus has infected over 509 million people worldwide and killed about 6.32 million up to date. Up on invasion, the Receptor Binding Domain (RBD) of Spike protein plays a crucial role in the entry of the virus into the host cell. The virus N protein is another protein that has a critical role for genome packaging. Methods: As bioinformatics approaches, the cassette design, codon adaptation, and protein stability were investigated in this study. Synthetic genes of RBD and N were cloned separately in pET28a + expression vector. They were transferred into Escherichia coli (E. coli) BL21 DE3 host cell, and expression of recombinant proteins was induced with IPTG. The recombinant proteins were purified by column chromatography and approved by Western blotting. Animal immunization was performed with each of the recombinant proteins individually and in combination of the two. The antibody titer of the blood serum from control and immunized mice groups was determined by ELISA technique. Finally, the anti-spike neutralization test was performed. Results: The expression and purification of RBD protein were monitored on SDS-PAGE, two bands of about 28 and 45 kDa for RBD and N appeared on gel distinctly, which were further validated by Western blotting. According to ELISA results, related antibodies were traced to a dilution of 1/64000 in immunized sera. The neutralization test exhibited produced antibodies' potency to bind the virus proteins. Using SPSS software, statistical analysis was performed by Duncan's test and T-test. Conclusion: According to the present study, recombinant proteins, either RBD alone or in combination with N adequately stimulated the immune response, and the raised antibodies could neutralize the virus in in vitro test.

Phyto-Fabrication of Silver Nanoparticles Using Typha azerbaijanensis Aerial Part and Root Extracts.

Background: Silver nanoparticles (AgNPs) were phyto-synthesized using Typha azerbaijanensis aerial part and root extracts, and their biological activities were investigated. Methods: This study was conducted in the Science and Research Branch, Islamic Azad University, Tehran, Iran in 2019. In this experimental study, silver nanoparticles (AgNPs) were phyto-synthesized and the physicochemical properties of AgNPs were determined using UV-Vis (UV-Vis) spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectroscopy. Antibacterial and anticancer activity of synthesized AgNPs was determined using microdilution assay, and MTT 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) methods, respectively. The apoptotic effects of AgNPs were investigated using Real-Time PCR and flow cytometry techniques. Results: Morphological analysis of the synthesized AgNPs confirmed the spherical shape of AgNPs with an average size of 10.67 to 16.69 nm. The FTIR spectrum confirmed the presence of phytochemicals from T. azerbayenensis extract at the AgNP surface. Antibacterial experiments showed that phyto-fabricated AgNPs had significant antibacterial activity against Gram-negative bacteria. The AgNPs were significantly cytotoxic against breast cancer cell line (MCF-7) through induction of apoptosis. Conclusion: The phyto-synthesized AgNPs had biological activities could be useful in pharmaceutical applications.

Inflammatory Markers in the Serum and Bronchoalveolar Lavage in Children with Non-Cystic Fibrosis Bronchiectasis.

Background: It is known that inflammatory responses occur in the airways of patients with non-cystic fibrosis bronchiectasis during respiratory exacerbations but the role of these cytokines is not clear in this condition. Herein we evaluated the levels of interleukin-1ß (IL-1ß), IL-8 and tumor necrosis factor α (TNF-α) in the serum and bronchoalveolar lavage among children with non-cystic fibrosis bronchiectasis. Materials and Methods: This cross-sectional study was performed on all children with non-cystic fibrosis bronchiectasis who were admitted with respiratory exacerbation in the pediatric pulmonology ward of Masih Daneshvari Hospital, Tehran-Iran. All patients underwent fiberoptic bronchoscopy and spirometry before and after the bronchoscopy. IL-1ß, IL-8, and TNF-α levels were measured in the serum and bronchoalveolar lavage. Results: Patients included 10 (59%) female and 7 (41%) male subjects with mean age of 13.8 years (range, 5-18). Mean values for forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) were below the normal range before and after bronchoscopy. Mean value for FVC (from 55% to 63%, P= 0.01) and FEV1 (from 60% to 64%, P= 0.26) increased after bronchoscopy compared to before that. IL-1ß and IL-8 levels were increased and TNF-α level was decreased in the serum and bronchoalveolar lavage but no significant correlation was found between spirometry and these cytokines levels. Conclusion: Changes in inflammatory cytokines levels in serum and bronchoalveolar lavage during respiratory exacerbation in patients with noncystic fibrosis bronchiectasis have no significant correlation with spirometry and cannot be used in clinical practice.

In vitro effects of silver nanoparticles on gills morphology of female Guppy (Poecilia reticulate) after a short-term exposure.

The application of nanoparticles in various industries has grown significantly in recent years. The aims of this study were evaluation effects of silver nanoparticles (AgNPs) on survival rate and tissues (Gills) of Guppy (Poecilia reticulate) as a model of the freshwater organism. For this purpose, 90 mature fish were exposed to a different level of AgNPs for 96 hr. Data analyzed showed there was a significant correlation between fish mortality rate and AgNPs concentrations. Histological assays showed some typical tissue damages such as hyperplasia, hypertrophy, hyperemia, hemorrhage, and necrosis. Moreover, fish exposed to a lethal concentration of AgNPs showed some clinical signs, such as increasing operculum movement, swimming near the water surface, anxiety, and death with open mouth. The results of the present study showed that AgNPs can have toxicity effects on Guppy, also to sub-lethal concentrations, leading to several tissue damages and reduced survival rate of fish.

Bronchiectasis in common variable immunodeficiency: A systematic review and meta-analysis.

BACKGROUND: Common variable immunodeficiency (CVID) is the most prevalent symptomatic primary immunodeficiency disorder characterized by infectious and noninfectious complications. Bronchiectasis continues to be a common respiratory problem and therapeutic challenge in CVID. The aim of this study is to estimate the overall prevalence of bronchiectasis and its associated phenotype in patients with CVID. METHODS: A systematic literature search was performed in Web of Science, PubMed, and Scopus from the earliest available date to February 2019 with standard keywords. All pooled analyses of bronchiectasis prevalence and the corresponding 95% confidence intervals (CIs) were based on random-effects models. RESULTS: Fifty-five studies comprising 8535 patients with CVID were included in the meta-analysis. Overall prevalence of bronchiectasis was 34% (95% CI: 30-38; I2 = 90.19%). CVID patients with bronchiectasis had significantly lower serum immunoglobulin A (IgA) and IgM levels at the time of diagnosis compared with those without bronchiectasis. Among the clinical features, the frequencies of splenomegaly, pneumonia, otitis media, and lymphocytic interstitial pneumonia were significantly higher in CVID patients with bronchiectasis compared with those without bronchiectasis, respectively. CONCLUSION: A higher prevalence of bronchiectasis in patients with CVID should be managed by controlling recurrent and severe pneumonia episodes which are immune dysregulation since this complication is associated with poor prognosis in these patients.

Approach for determination of detonation performance and aluminum percentage of aluminized-based explosives by laser-induced breakdown spectroscopy.

Energetic materials containing aluminum powder are hazardous compounds, which have wide applications as propellants, explosives, and pyrotechnics. This work introduces a new method on the basis of the laser-induced breakdown spectroscopy technique in air and argon atmospheres to investigate determination of aluminum content and detonation performance of 1,3,5-trinitro-1,3,5-triazine (RDX)-based aluminized explosives. Plasma emission of aluminized RDX explosives are recorded where atomic lines of Al, C, H, N, and O, as well as molecular bands of AlO and CN are identified. The formation mechanism of AlO and CN molecular bands is affected by the aluminum percentage and oxygen content present in the composition and plasma. Relative intensity of the Al/O is used to determine detonation velocity and pressure of the RDX/Al samples. The released energy in the laser-induced plasma of aluminized RDX composition is related to the heat of explosion and percentage of aluminum.

Environmental impact assessment of the industrial estate development plan with the geographical information system and matrix methods.

BACKGROUND: The purpose of this study is environmental impact assessment of the industrial estate development planning. METHODS: This cross-sectional study was conducted in 2010 in Isfahan province, Iran. GIS and matrix methods were applied. Data analysis was done to identify the current situation of the region, zoning vulnerable areas, and scoping the region. Quantitative evaluation was done by using matrix of Wooten and Rau. RESULTS: The net score for impact of industrial units operation on air quality of the project area was (-3). According to the transition of industrial estate pollutants, residential places located in the radius of 2500 meters of the city were expected to be affected more. The net score for impact of construction of industrial units on plant species of the project area was (-2). Environmental protected areas were not affected by the air and soil pollutants because of their distance from industrial estate. CONCLUSION: Positive effects of project activities outweigh the drawbacks and the sum scores allocated to the project activities on environmental factor was (+37). Totally it does not have detrimental effects on the environment and residential neighborhood. EIA should be considered as an anticipatory, participatory environmental management tool before determining a plan application.

Juvenile churg-strauss syndrome as an etiology of myocarditis and ischemic stroke in adolescents; a case report.

BACKGROUND: Churg-Strauss syndrome (CSS), a systemic vasculitis accompanied by asthma and eosinophilia, almost invariably affects the lung and is frequently associated with cutaneous involvement. It rarely has cardiac involvement. We report an unusual case of CSS with myocardial involvement and stroke. CASE PRESENTATION: A 16-year old female suffered of allergic asthma for 4 years. She was under treatment with oral prednisolone and seretide inhalation. After CSS diagnosis, she developed paroxysmal atrial tachycardia. Serum levels of Troponin I and Troponin T were increased indicating massive myocardial damage probably due to myocarditis. After 5 months she developed acute hemiparesis without any evidence of ischemic or hemorrhagic event. She was treated with IVIg, intravenous pulses of methylprednisone and cyclophosphamide for each complication. CONCLUSION: Myocarditis and stroke may also complicate CSS which should be taken in consideration for better management.

Characterization of 11 new cases of leukocyte adhesion deficiency type 1 with seven novel mutations in the ITGB2 gene.

BACKGROUND: Leukocyte adhesion deficiency type 1 (LAD I) is an autosomal recessive disorder caused by mutations in the ITGB2 gene, encoding the beta2 integrin family. Severe recurrent infections, impaired wound healing, and periodontal diseases are the main features of disease. METHODS: In order to investigate clinical and molecular manifestations of new LAD I cases, 11 patients diagnosed in one center during 7 years were studied. Patients were screened for the ITGB2 gene mutations, using polymerase chain reaction, followed by single-strand conformation polymorphism and sequencing. RESULTS: The most common first presenting feature of the patients was omphalitis. The mean age of cord separation was 19.9 +/- 1 days. The most common clinical manifestations of the patients during the follow-up period included omphalitis, skin ulcers with poor healing, sepsis, and otitis media. During the follow-up, eight patients died. Eight homozygous changes, including seven novel mutations, were detected: two splicing (IVS4-6C>A, IVS7+1G>A), three missense (Asp128Tyr, Ala239Thr, and Gly716Ala), and three frameshift deletions (Asn282fsX41, Tyr382fsX9, and Lys636fsX22). CONCLUSION: Our results indicate that different mutations underlie the development of LAD I. Definitive molecular diagnosis is valuable for genetic counseling and prenatal diagnosis. Regarding clinical presentations, it seems that omphalitis is the most consistent finding seen in LAD I infants.

Spectrophotometric study of acidity constants of Alizarine Red S in various water-organic solvent.

The acidity constants of Alizarine Red S were determined spectrophotometrically at 25 degrees C and at constant ionic strength 0.1 M (KNO3) in pure water as well as in aqueous media containing variable mole percentages (5-70%) of organic solvents. The organic solvents used were methanol, ethanol, N,N-dimethyl formamide (DMF), dimethyl sulfoxide (DMSO), acetonitrile and dioxan. The acidity constants of all related equilibria are estimated using the whole spectral fitting of the collected data to an established factor analysis model. DATAN program was applied for determining of acidity constants and pure spectra of different form of Alizarine Red S. The obtained results indicated that acidity constants decrease as the content of an organic solvent in the medium increases. There are linear relationship between acidity constants and the mole fraction of various organic solvents in the solvent mixtures. Effect of various solvents on acidity constants and pure spectrum of each component are also discussed.

Pentoxifyllin attenuates the systemic inflammatory response induced during isolated limb perfusion with recombinant human tumor necrosis factor-alpha and melphalan.

BACKGROUND: Isolated limb perfusion (ILP) with recombinant human tumor necrosis factor-alpha (rhTNF-alpha) and melphalan harbors the risk of septic shock-like syndrome. Pentoxifyllin (PTX) produced a beneficial effect on cytokine response and survival in animal experiments of septic shock, and we were interested to explore its effect during TNF-ILP in humans. METHODS: Eighteen consecutive patients underwent TNF-ILP and received PTX (30 mg/kg/day), whereas another 13 consecutive patients did not. PTX was given systemically after the limb extracorporeal circulation was started. Cardiac index, systemic vascular resistance (SVR), and pulmonary vascular resistance were recorded via a Swan-Ganz catheter. Blood levels of TNF-alpha, interleukin-6, procalcitonin, and lipopolysaccharide-binding protein were determined before, during, and after ILP. RESULTS: After reperfusion, systemic levels of TNF-alpha were significantly less increased in the PTX group (peak, 2.8 vs. 1.3 ng/mL; P <.05), as were interleukin-6 values (peak, 68 vs. 22 pg/mL; P <.02) and lipopolysaccharide-binding protein plasma levels (peak, 215 vs. 105 micro g/mL; P <.03). Differences in cardiac index, SVR, and mean arterial blood pressure were not significantly different. Norepinephrine or dobutamine to maintain SVR was less required in the PTX group. CONCLUSIONS: PTX attenuates systemic cytokine production and influences components of the systemic inflammatory response after TNF-ILP. PTX may play a beneficial role in the management of septic shock-like syndrome, particularly in patients with leakage from the ILP circuit.