ResearchPad - research-letters https://www.researchpad.co Default RSS Feed en-us © 2020 Newgen KnowledgeWorks <![CDATA[The COVID-19 outbreak and the angiotensin-converting enzyme 2: too little or too much?]]> https://www.researchpad.co/article/N847ad536-bd17-4faf-9ae8-81eeba67ca42 <![CDATA[Fosfomycin suppresses RS-virus-induced Streptococcus pneumoniae and Haemophilus influenzae adhesion to respiratory epithelial cells via the platelet-activating factor receptor]]> https://www.researchpad.co/article/N42e72f66-9f12-4426-a63b-3905d2c1b046

Abstract

Human respiratory syncytial virus (RSV) sometimes causes acute and severe lower respiratory tract illness in infants and young children. The platelet-activating factor (PAF) receptor, which is a receptor for Streptococcus pneumoniae and Haemophilus influenzae, is upregulated by RSV infection in the pulmonary epithelial cell line A549. Fosfomycin, an antimicrobial agent, significantly suppressed PAF receptor induction by RSV infection at the mRNA and cell surface expression levels. Fosfomycin also suppressed RSV-induced adhesion of fluorescence-labeled S. pneumoniae and H. influenzae cells, as determined by flow cytometry and fluorescence microscopy. The RSV-induced bacterial adhesion was suggested to be host-PAF-receptor and bacterial-phosphocholine mediated. Fosfomycin, which has been shown to exhibit antimicrobial and immunomodulatory activities, was found here to suppress adhesion by disease-causing bacteria. Thus, fosfomycin might prevent secondary bacterial infection during RSV infection.

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<![CDATA[Human Bocavirus in Very Young Infants Hospitalized with Acute Respiratory Infection in Northeast Brazil]]> https://www.researchpad.co/article/Ncfba4a31-f104-4e3f-a0e4-a4e2f3745cda

Abstract

A cross-sectional study was carried out over a period of 12 months to investigate the occurrence of human bocavirus (HBoV) infection in infants hospitalized for respiratory infections in a teaching hospital in Salvador, Brazil, and to describe the clinical manifestations of this infection. Nasopharyngeal aspirates were collected from the children and immunofluorescence and polymerase chain reaction were performed to investigate the presence of respiratory viruses. HBoV was detected in 4 out of 66 patients. Two of the HBoV-positive infants were co-infected with other viruses. The principal clinical findings in HBoV-positive children were: nasal obstruction, catarrh, cough, fever and dyspnea. This study revealed HBoV infection in children aged <2 months, suggesting that the infection may occur at a very early age.

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<![CDATA[Human G-CSF synthesis using stress-responsive bacterial proteins]]> https://www.researchpad.co/article/Nb6167d58-7b03-4b50-859c-a2c9f5855aee

Abstract

We previously reported that under the stress condition caused by the addition of 2-hydroxyethyl disulfide, a thiol-specific oxidant, to growing cultures of Escherichia coli BL21(DE3), a population of stress-responsive proteins [peptidyl-prolyl cistrans isomerase B (PpiB), bacterioferritin (Bfr), putative HTH-type transcriptional regulator yjdC (YjdC), dihydrofolate reductase (FolA), chemotaxis protein cheZ (CheZ), and glutathione synthetase (GshB)] were significantly upregulated when compared with the nonstress condition. When those stress-responsive proteins were used as fusion partners for the expression of human granulocyte colony-stimulating factor (hG-CSF), the solubility of hG-CSF was dramatically enhanced in E. coli cytoplasm, whereas almost all of the directly expressed hG-CSF were aggregated to inclusion bodies. In addition, the spectra of circular dichroism measured with the purified hG-CSF were identical to that of standard hG-CSF, implying that the synthesized hG-CSF has native conformation. These results indicate that the bacterial stress-responsive proteins could be potent fusion expression partners for aggregation-prone heterologous proteins in E. coli cytoplasm.

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<![CDATA[Occurrence of enterotoxigenic and nonenterotoxigenic Bacteroides fragilis in calves and evaluation of their antimicrobial susceptibility]]> https://www.researchpad.co/article/N6f6e665b-c0e5-42a3-be70-6048ff305774

Abstract

Bacteroides fragilis is considered an important clinical pathogen and the most common anaerobe isolated from human and animal clinical specimens; enterotoxigenic strains produce diarrhea. The presence of enterotoxigenic (ETBF) and nonenterotoxigenic B. fragilis in stool samples from calves with or without acute diarrhea and the antimicrobial susceptibility of the strains were evaluated. The stool samples were plated onto a selective B. fragilis–bile–esculin agar, and incubated anaerobically (10% CO2/90% N2), at 37°C, for 72 h. Species of the B. fragilis group were identified by using the API 32‐A kit. Enterotoxigenic strains were detected by PCR and the cytotoxic assay. From 54 diarrhea and 54 nondiarrhea stools, 124 and 92 members of the B. fragilis group, respectively, were recovered. Only two ETBF strains were isolated from two different diarrhea samples and the bft gene was detected in both. Moreover, the bft gene was detected in DNA from four different diarrheal stools samples but no ETBF strain was recovered. All the bacteria were susceptible to chloramphenicol, imipenem, moxifloxacin, piperacillin/tazobactam, metronidazole and tigecycline. Most of the isolates from both calves with and without diarrhea were resistant to all metals. Our results are of concern, and suggest the need to increase the surveillance of antibiotic and metal resistance of this microbial group isolated from animal production such as calves.

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<![CDATA[Loop-mediated isothermal amplification method for rapid detection of the toxic dinoflagellate Alexandrium, which causes algal blooms and poisoning of shellfish]]> https://www.researchpad.co/article/N11078a7a-149f-4a2b-8d46-9a3ad8dd01a7

Abstract

The marine dinoflagellate genus Alexandrium includes a number of species that produce potent neurotoxins responsible for paralytic shellfish poisoning, which in humans may cause muscular paralysis, neurological symptoms and, in extreme cases, death. Because of the genetic diversity of different genera and species, molecular tools may help to detect the presence of target microorganisms in marine field samples. Here we employed a loop-mediated isothermal amplification (LAMP) method for the rapid and simple detection of toxic Alexandrium species. A set of four primers were designed based upon the conserved region of the 5.8S rRNA gene of members of the genus Alexandrium. Using this detection system, toxic Alexandrium genes were amplified and visualized as a ladder-like pattern of bands on agarose gels under isothermal condition within 60 min. The LAMP amplicons were also directly visualized by eye in the reaction tube by the addition of SYBR Green I. This LAMP assay was 10-fold more sensitive than a conventional PCR method with a detection limit of 5 cells per tube when targeting DNA from Alexandrium minutum. The LAMP assay reported here indicates the potential usefulness of the technique as a valuable simple, rapid alternative procedure for the detection of target toxic Alexandrium species during coastal water monitoring.

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<![CDATA[Identification of Mycobacterium species by comparative analysis of the dnaA gene]]> https://www.researchpad.co/article/N6e82d0cb-a8e5-4824-ae62-8b65b3908a62

Abstract

For the establishment of a diagnostic tool for mycobacterial species, a part of the dnaA gene was amplified and sequenced from clinically relevant 27 mycobacterial species as well as 49 clinical isolates. Sequence variability in the amplified segment of the dnaA gene allowed the differentiation of all species except for Mycobacterium tuberculosis, Mycobacterium africanum and Mycobacterium microti, which had identical sequences. Partial sequences of dnaA from clinical isolates belonging to three frequently isolated species revealed a very high intraspecies similarity, with a range of 96.0–100%. Based on the dnaA sequences, a species-specific primer set for Mycobacterium kansasii and Mycobacterium gastri was successfully designed for a simple loop-mediated isothermal amplification method. These results demonstrate that the variable sequences in the dnaA gene were species specific and were sufficient for the development of an accurate and rapid diagnosis of Mycobacterium species.

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<![CDATA[Existence and characterization of allelic variants of Sao, a newly identified surface protein from Streptococcus suis]]> https://www.researchpad.co/article/Ne9cd5b0c-077b-4d33-999f-4e02e380da8d

Abstract

Surface antigen one (Sao) is a newly identified protein from the major zoonotic pathogen, Streptococcus suis. In search of functional proteins related to the pathogenesis of Chinese S. suis 2 (SS2), unexpectedly, a variant of Sao protein was obtained. To test its prevalence in S. suis, PCR assay was adopted to address the coding genes systematically. It was found that there are three allelic variants of sao gene, namely sao-S, sao-M, and sao-L based on the different lengths of the genes (∼1.5, ∼1.7, and ∼2.0 kb, respectively). These differences were determined to be caused by heterogeneity within the number of C-terminal repeat sequences (R), which had been seen as a pathogenicity-related domain in the plant pathogen, Xanthomonas oryzae. Two variants (sao-M and sao-L) were only found in SS2. All three variant proteins were prepared in vitro and their biochemical and biophysical properties were characterized. A soluble form of Sao-M protein was then used as a capture antigen to develop an enzyme-linked immunosorbent assay method to detect antibodies against SS2 in convalescent pig sera. Taken together, the results exhibit the properties of Sao proteins and provide an efficient Sao-M-based method for monitoring SS2 infection.

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<![CDATA[Secretion of M2e:HBc fusion protein by Lactobacillus casei using Cwh signal peptide]]> https://www.researchpad.co/article/Nbca377d5-dc57-486f-828c-704a23970efa

The ability to serve as a delivery vehicle for various interesting biomolecules makes lactic acid bacteria (LAB) very useful in several applications. In the medical field, recombinant LAB expressing pathogenic antigens at different cellular locations have been used to elicit both mucosal and systemic immune responses. Expression–secretion vectors (ESVs) with a signal peptide (SP) are pivotal for protein expression and secretion. In this study, the genome sequence of Lactobacillus casei ATCC334 was explored for new SPs using bioinformatics tools. Three new SPs of the proteins Cwh, SurA and SP6565 were identified and used to construct an ESV based on our Escherichia coli–L. casei shuttle vector, pRCEID-LC13.9. Functional testing of these constructs with the green fluorescence protein (GFP) gene showed that they could secrete the GFP. The construct with CwhSP showed the highest GFP secretion. Consequently, CwhSP was selected to develop an ESV construct carrying a synthetic gene encoding the extracellular domain of the matrix 2 protein fused with the hepatitis B core antigen (M2e:HBc). This ESV was shown to efficiently express and secrete the M2e:HBc fusion protein. The identified SPs and the developed ESVs can be exploited for expression and secretion of homologous and heterologous proteins in L. casei.

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<![CDATA[Evaluation of P1 adhesin epitopes for the serodiagnosis of Mycoplasma pneumoniae infections]]> https://www.researchpad.co/article/N1f7d1888-38f2-4d0f-a618-2fccfb501a93

Abstract

Most glycolipid antigens used for serological tests of Mycoplasma pneumoniae are not M. pneumonia‐specific, and can cross‐react with other microorganism antigens and body tissues, resulting in false positives. It is important to identify M. pneumonia‐specific antigen(s) for serological testing and correct diagnosis. Two epitopes, rP1‐534 and rP1‐513, of P1 adhesin predicted by bioinformatics were successfully expressed and purified, and could be recognized by serum samples from M. pneumoniae‐infected patients and His tag antibodies by Western blot. There was no cross‐reactivity between the anti‐recombinant proteins serum and other respiratory antigens. A total of 400 patients were investigated, their respiratory specimens tested by PCR, and sera tested by a commercial test kit; 56 with positive sera and positive respiratory specimens were designated as standard positive serum and 63 patients were designated as standard negative serum. The purified recombinant proteins were used as a combination of antigens or separately to test the serum. Serological test demonstrated that rP1‐513 of the C terminal of P1 adhesin is a new candidate antigen with greater sensitivity and specificity for IgG and IgM serodiagnosis of M. pneumoniae‐infected patients. The results confirmed that rP1‐513 could be a useful new antigen for the immunodiagnosis of M. pneumoniae infection.

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<![CDATA[A conserved OmpA-like protein in Legionella pneumophila required for efficient intracellular replication]]> https://www.researchpad.co/article/Nee61258f-4d6e-4ba5-bf69-4b0f4aa1ddee

The OmpA-like protein domain has been associated with peptidoglycan-binding proteins, and is often found in virulence factors of bacterial pathogens. The intracellular pathogen Legionella pneumophila encodes for six proteins that contain the OmpA-like domain, among them the highly conserved uncharacterized protein we named CmpA. Here we set out to characterize the CmpA protein and determine its contribution to intracellular survival of L. pneumophila. Secondary structure analysis suggests that CmpA is an inner membrane protein with a peptidoglycan-binding domain at the C-teminus. A cmpA mutant was able to replicate normally in broth, but failed to compete with an isogenic wild-type strain in an intracellular growth competition assay. The cmpA mutant also displayed significant intracellular growth defects in both the protozoan host Acanthamoeba castellanii and in primary bone marrow-derived macrophages, where uptake into the cells was also impaired. The cmpA phenotypes were completely restored upon expression of CmpA in trans. The data presented here establish CmpA as a novel virulence factor of L. pneumophila that is required for efficient intracellular replication in both mammalian and protozoan hosts.

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<![CDATA[Comparison of expression vectors in Lactobacillus reuteri strains]]> https://www.researchpad.co/article/Na3d975b8-fb8e-489a-b96f-a4f02639cd41

Abstract

The synthesis of heterologous proteins in lactobacilli is strongly influenced by the promoter selected for the expression. In addition, the activity of the promoters themselves may vary among different bacterial hosts. Three different promoters were investigated for their capability to drive enhanced green fluorescent protein (EGFP) expression in Lactococcus lactis spp. cremoris MG1363, in Lactobacillus reuteri DSM 20016T and in five L. reuteri strains isolated from chicken crops. The promoters of the Lactobacillus acidophilus surface layer protein gene (slp), L. acidophilus lactate dehydrogenase gene (ldhL) and enterococcal rRNA adenine N-6-methyltransferase gene (ermB) were fused to the coding sequence of EGFP and inserted into the backbone of the pTRKH3 shuttle vector (pTRKH3-slpGFP, pTRKH3-ldhGFP, pTRKH3-ermGFP). Besides conventional analytical methods, a new quick fluorimetric approach was set up to quantify the EGFP fluorescence in transformed clones using the Qubit fluorometer. ermB proved to be the most effective promoter in L. reuteri isolates, producing 3.90 × 10−7 g of fluorescent EGFP (mL ODstationary culture)−1. Under the same conditions, the ldhL promoter produced 2.66 × 10−7 g of fluorescent EGFP (mL ODstationary culture)−1. Even though the slp promoter was efficient in L. lactis spp. cremoris MG1363, it was nearly inactive both in L. reuteri DSM 20016T and in L. reuteri isolates.

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<![CDATA[Is pulmonary arterial hypertension associated with interferon-β therapy for multiple sclerosis reversible? A case study to explore the complexity]]> https://www.researchpad.co/article/N8f301b8a-50e4-4f08-83a8-04f8a872601b

Interferon (IFN)-β has been classified as a drug that is possibly associated with development of pulmonary arterial hypertension (PAH), a devastating disease that can lead to right heart failure and premature death [1].

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<![CDATA[Nonpharmacological cough control therapy for chronic refractory cough and cough associated with underlying lung disease]]> https://www.researchpad.co/article/N4a8a8b86-424d-4ae8-87f8-ab1f4f99fe8a

Cough is a common and frequently debilitating symptom. A persistent cough unresponsive to empirical treatment (chronic refractory cough (CRC)) or unexplained may represent a distinct phenotype [1]. The pathophysiology of CRC is poorly understood. “Cough hypersensitivity syndrome” has recently been the dominant paradigm. Evidence suggests a complex interplay of aberrant vagal afferent pathways [2] and central factors including impaired cough suppression [3]. Cough is more usually associated with an underlying cause (explained cough) and may persist in patients with underlying lung disease even after optimising treatment of the condition. Cough reflex hypersensitivity may be a specific treatable trait in airway disease [4]. Cough is common in asthma; studies demonstrate a link between airway dysfunction and cough hypersensitivity [5]. There are similar findings in bronchiectasis [6] and COPD [7], although the underlying mechanisms may differ.

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<![CDATA[Tau (297‐391) forms filaments that structurally mimic the core of paired helical filaments in Alzheimer’s disease brain]]> https://www.researchpad.co/article/Nc26f9bfb-6a6e-48db-8fd8-8cea19285000

The constituent paired helical filaments (PHFs) in neurofibrillary tangles are insoluble intracellular deposits central to the development of Alzheimer’s disease (AD) and other tauopathies. Full‐length tau requires the addition of anionic cofactors such as heparin to enhance assembly. We have shown that a fragment from the proteolytically stable core of the PHF, tau 297‐391 known as ‘dGAE’, spontaneously forms cross‐β‐containing PHFs and straight filaments under physiological conditions. Here, we have analysed and compared the structures of the filaments formed by dGAE in vitro with those deposited in the brains of individuals diagnosed with AD. We show that dGAE forms PHFs that share a macromolecular structure similar to those found in brain tissue. Thus, dGAEs may serve as a model system for studying core domain assembly and for screening for inhibitors of tau aggregation.

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<![CDATA[A missense mutation in the catalytic domain of O‐GlcNAc transferase links perturbations in protein O‐GlcNAcylation to X‐linked intellectual disability]]> https://www.researchpad.co/article/N4cff86d7-f6c8-48a5-b3fb-920d0e162958

X‐linked intellectual disabilities (XLID) are common developmental disorders. The enzyme O‐GlcNAc transferase encoded by OGT, a recently discovered XLID gene, attaches O‐GlcNAc to nuclear and cytoplasmic proteins. As few missense mutations have been described, it is unclear what the aetiology of the patient phenotypes is. Here, we report the discovery of a missense mutation in the catalytic domain of OGT in an XLID patient. X‐ray crystallography reveals that this variant leads to structural rearrangements in the catalytic domain. The mutation reduces in vitro OGT activity on substrate peptides/protein. Mouse embryonic stem cells carrying the mutation reveal reduced O‐GlcNAcase (OGA) and global O‐GlcNAc levels. These data suggest a direct link between changes in the O‐GlcNAcome and intellectual disability observed in patients carrying OGT mutations.

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<![CDATA[Magma Degassing as a Source of Long‐Term Seismicity at Volcanoes: The Ischia Island (Italy) Case]]> https://www.researchpad.co/article/N978e9b20-749d-4a9f-ac68-8956762af4c2

Abstract

Transient seismicity at active volcanoes poses a significant risk in addition to eruptive activity. This risk is powered by the common belief that volcanic seismicity cannot be forecast, even on a long term. Here we investigate the nature of volcanic seismicity to try to improve our forecasting capacity. To this aim, we consider Ischia volcano (Italy), which suffered similar earthquakes along its uplifted resurgent block. We show that this seismicity marks an acceleration of decades‐long subsidence of the resurgent block, driven by degassing of magma that previously produced the uplift, a process not observed at other volcanoes. Degassing will continue for hundreds to thousands of years, causing protracted seismicity and will likely be accompanied by moderate and damaging earthquakes. The possibility to constrain the future duration of seismicity at Ischia indicates that our capacity to forecast earthquakes might be enhanced when seismic activity results from long‐term magmatic processes, such as degassing

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<![CDATA[LAMTOR/Ragulator regulates lipid metabolism in macrophages and foam cell differentiation]]> https://www.researchpad.co/article/N972e3e0b-40a3-46ff-8be1-ca6fc17b3d6d

Late endosomal/lysosomal adaptor and MAPK and mTOR activator (LAMTOR/Ragulator) is a scaffold protein complex that anchors and regulates multiprotein signaling units on late endosomes/lysosomes. To identify LAMTOR‐modulated endolysosomal proteins, primary macrophages were derived from bone marrow of conditional knockout mice carrying a specific deletion of LAMTOR2 in the monocyte/macrophage cell lineage. Affymetrix‐based transcriptomic analysis and quantitative iTRAQ‐based organelle proteomic analysis of endosomes derived from macrophages were performed. Further analyses showed that LAMTOR could be a novel regulator of foam cell differentiation. The lipid droplet formation phenotype observed in macrophages was additionally confirmed in MEFs, where lipidomic analysis identified cholesterol esters as specifically downregulated in LAMTOR2 knockout cells. The data obtained indicate a function of LAMTOR2 in lipid metabolism.

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<![CDATA[Increasing mortality rate due to rheumatoid arthritis-related lung diseases in Japan]]> https://www.researchpad.co/article/N3efdaab2-fd8a-429b-9edd-6e0892489812

The quality of life of rheumatoid arthritis (RA) patients has dramatically improved in the approximately two decades since biological agents were first introduced for its treatment [1]. However, whether current treatment strategies for RA have helped improve the prognosis of patients suffering from complications due to RA remains controversial. National data from the USA have shown that the mortality rate of RA-associated interstitial lung disease (ILD), a major complication associated with a poor prognosis in RA patients, seems to be gradually increasing, whereas the mortality rate of patients with any type of RA-associated disease has been decreasing [2].

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<![CDATA[Quantifying the Timescale and Strength of Southern Hemisphere Intraseasonal Stratosphere‐troposphere Coupling]]> https://www.researchpad.co/article/Ne2d0b07a-2452-4f6d-ba26-c3d3b5569795

Abstract

The Southern Hemisphere zonal circulation manifests a downward influence of the stratosphere on the troposphere from late spring to early summer. However, the strength and timescale of the connection, given the stratospheric state, have not been explicitly quantified. Here, SH zonal wind reanalysis time series are analyzed with a methodology designed to detect the minimal set of statistical predictors of multiple interacting variables via conditional independence tests. Our results confirm from data that the variability of the stratospheric polar vortex is a predictor of the tropospheric eddy‐driven jet between September and January. The vortex variability explains about 40% of monthly mean jet variability at a lead time of 1 month and can entirely account for the observed jet persistence. Our statistical model can quantitatively connect the multidecadal trends observed in the vortex and jet during the satellite era. This shows how short‐term variability can help understand statistical links in long‐term changes.

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