A Viewpoint Commentary: Buffalo as a New Model for Long COVID-19 Study and for Recovering the SARS CoV-2 Polyclonal Neutralizing Antibody, Using an Online Affinity Column Adsorption Technology, for Therapeutic Interventional Modality in Humans: A New Mult

Understanding in population structure of a plant’s root-associated microbiome is applied in good practices in agricultural activities to improve production yields and enhance plant immune responses. The molecular analysis of bacterial populations inhabited in soil faces difficulties in obtaining high yield and high purity of DNA, and different commercial DNA extraction kits have been developed for this purpose. This study focuses on the comparison of DNA extraction of six different rice root-associated bacterial consortium using three commercial kits with two key technologies, spin-column adsorption and magnetic bead adsorption. The quality and quantity of genomic DNA obtained from these extraction methods were analyzed and compared based on DNA concentration, DNA purity and efficiency to be used as a template for 16sRNA amplification. The results showed that the extraction kit with magnetic bead adsorption technology showed the highest concentration (101.32 ng/μl) compared to other DNA extraction kits (32.67 and 1.89 ng/μl). The purity values (A260/A280) were assessed by using Nano-drop spectrophotometer and resulted in purities of nucleic acids in the range of 1.4-1.7. Thus, it was concluded that the extracted DNA obtained from the extraction kit with magnetic bead adsorption technology can be valuable for molecular analysis of microbial communities present in the soil.

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SARS-CoV-2 Proteins Bind to Hemoglobin and Its Metabolites

International Journal of Molecular Sciences ◽

10.3390/ijms22169035 ◽

2021 ◽

Vol 22 (16) ◽

pp. 9035

Author(s):

Guilherme C. Lechuga ◽

Franklin Souza-Silva ◽

Carolina Q. Sacramento ◽

Monique R. O. Trugilho ◽

Richard H. Valente ◽

...

Keyword(s):

In Silico ◽

Plaque Assay ◽

In Silico Analysis ◽

Inhibitory Effect ◽

Parallel Synthesis ◽

Affinity Column ◽

Binding Motifs ◽

N Protein ◽

Column Adsorption ◽

Silico Analysis

(1) Background: coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been linked to hematological dysfunctions, but there are little experimental data that explain this. Spike (S) and Nucleoprotein (N) proteins have been putatively associated with these dysfunctions. In this work, we analyzed the recruitment of hemoglobin (Hb) and other metabolites (hemin and protoporphyrin IX-PpIX) by SARS-Cov2 proteins using different approaches. (2) Methods: shotgun proteomics (LC–MS/MS) after affinity column adsorption identified hemin-binding SARS-CoV-2 proteins. The parallel synthesis of the peptides technique was used to study the interaction of the receptor bind domain (RBD) and N-terminal domain (NTD) of the S protein with Hb and in silico analysis to identify the binding motifs of the N protein. The plaque assay was used to investigate the inhibitory effect of Hb and the metabolites hemin and PpIX on virus adsorption and replication in Vero cells. (3) Results: the proteomic analysis by LC–MS/MS identified the S, N, M, Nsp3, and Nsp7 as putative hemin-binding proteins. Six short sequences in the RBD and 11 in the NTD of the spike were identified by microarray of peptides to interact with Hb and tree motifs in the N protein by in silico analysis to bind with heme. An inhibitory effect in vitro of Hb, hemin, and PpIX at different levels was observed. Strikingly, free Hb at 1mM suppressed viral replication (99%), and its interaction with SARS-CoV-2 was localized into the RBD region of the spike protein. (4) Conclusions: in this study, we identified that (at least) five proteins (S, N, M, Nsp3, and Nsp7) of SARS-CoV-2 recruit Hb/metabolites. The motifs of the RDB of SARS-CoV-2 spike, which binds Hb, and the sites of the heme bind-N protein were disclosed. In addition, these compounds and PpIX block the virus’s adsorption and replication. Furthermore, we also identified heme-binding motifs and interaction with hemin in N protein and other structural (S and M) and non-structural (Nsp3 and Nsp7) proteins.

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The Hitachi Model HS-8 Electron Microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100061203 ◽

1967 ◽

Vol 25 ◽

pp. 238-239

Author(s):

H. Akabori ◽

K. Nishiwaki ◽

K. Yoneta

Keyword(s):

Electron Microscope ◽

New Model ◽

Predecessor Model

By improving the predecessor Model HS- 7 electron microscope for the purpose of easier operation, we have recently completed new Model HS-8 electron microscope featuring higher performance and ease of operation.

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Presence of antibody in virus-infected brain cells of SSPE ferrets

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100077591 ◽

1983 ◽

Vol 41 ◽

pp. 804-805

Author(s):

Hannah R. Brown ◽

Tammy L. Donato ◽

Halldor Thormar

Keyword(s):

Measles Virus ◽

Plasma Cells ◽

Subacute Sclerosing Panencephalitis ◽

Protein A ◽

Neutralizing Antibody ◽

Brain Cells ◽

Antibody Titers ◽

A Cell ◽

The Central Nervous System ◽

The Brain

Measles virus specific immunoglobulin G (IgG) has been found in the brains of patients with subacute sclerosing panencephalitis (SSPE), a slowly progressing disease of the central nervous system (CNS) in children. IgG/albumin ratios indicate that the antibodies are synthesized within the CNS. Using the ferret as an animal model to study the disease, we have been attempting to localize the Ig's in the brains of animals inoculated with a cell associated strain of SSPE. In an earlier report, preliminary results using Protein A conjugated to horseradish peroxidase (PrAPx) (Dynatech Diagnostics Inc., South Windham, ME.) to detect antibodies revealed the presence of immunoglobulin mainly in antibody-producing plasma cells in inflammatory lesions and not in infected brain cells.In the present experiment we studied the brain of an SSPE ferret with neutralizing antibody titers of 1:1024 in serum and 1:512 in CSF at time of sacrifice 7 months after i.c. inoculation with SSPE measles virus-infected cells. The animal was perfused with saline and portions of the brain and spinal cord were immersed in periodate-lysine-paraformaldehyde (P-L-P) fixative. The ferret was not perfused with fixative because parts of the brain were used for virus isolation.

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Immunocytochemical localization of β1-4 galactosyltransferase in endometrial carcinoma cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162284 ◽

1990 ◽

Vol 48 (3) ◽

pp. 944-945

Author(s):

Ichiro Yamamoto ◽

Toshiaki Tachibana ◽

Hiroko Maruyama ◽

Noriyuki Komatsu ◽

Hiroyuki Kuramoto ◽

...

Keyword(s):

Endometrial Carcinoma ◽

Cell Lines ◽

Endometrial Adenocarcinoma ◽

Protein A ◽

Rabbit Antiserum ◽

Carcinoma Cells ◽

Affinity Column ◽

Antibody Technique ◽

Galactosyl Transferase ◽

C Terminus

We have paid attention to the alteration of glycosyltransferase in carcinoma cells, because it might be related to the malignancy of the cells. In this connection, localization of β1-4 galactosyl transferase (β1-4 Gal T) in human endometrial carcinoma cells was examined immunocytochemically using two kinds of cell lines, each of which showed different degree of differentiation.An antibody was purified from the rabbit antiserum against the synthetic peptide, IFNRLVFRGMSC (W89) of human β1-4 Gal T coupled with KLH (keyhole limpet hemocyanine) by protein A column and peptide-affinity column chromatography. The anti-W89 serum reacts to the C-terminus of human β 1-4 Gal T and to both membrane-bound and soluble forms of the enzyme. Cell line of well differentiated endometrial adenocarcinoma (I) and that of poorly differentiated endometrial adenocarcinoma (50B) were cultivated respectively in MEM medium containing 15% FCS and 2 mM glutamine for 4 d at 37°C under 5% CO2. The cells were fixed in a mixture of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M Soerensen’s phosphate buffer (pH 7.4) at 4°C for 30 min, washed with PBS, then freezed and thawed. The indirect method of the peroxidase- labeled antibody technique was used for immunocytochemistry of both LM and TEM on the cell lines. The cells were dehydrated in ethanol and embedded in TAAB 812. Ultrathin sections were observed under a TEM, JEM-100S.

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516: A Simplified Method to Estimate the Absolute Renal Uptake of 99MTC-DMSA: Evaluation with a New Model using the Radioactivity of Nephrectomy Specimens as Reference

The Journal of Urology ◽

10.1016/s0022-5347(18)34756-6 ◽

2005 ◽

Vol 173 (4S) ◽

pp. 140-141

Author(s):

Mariana Lima ◽

Celso D. Ramos ◽

Sérgio Q. Brunetto ◽

Marcelo Lopes de Lima ◽

Carla R.M. Sansana ◽

...

Keyword(s):

New Model ◽

Renal Uptake ◽

Simplified Method ◽

The Absolute

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476 Chronic left atrial volume and pressure overload in the goat: a new model of sustained atrial fibrillation

EP Europace ◽

10.1016/s1099-5129(05)80314-9 ◽

2005 ◽

Vol 7 ◽

pp. 107-107

Keyword(s):

Atrial Fibrillation ◽

Left Atrial ◽

Pressure Overload ◽

Left Atrial Volume ◽

Atrial Volume ◽

New Model

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Analyzing Stochastic Dependence of Cognitive Processes in Multidimensional Source Recognition

Experimental Psychology (formerly Zeitschrift für Experimentelle Psychologie) ◽

10.1027/1618-3169/a000261 ◽

2014 ◽

Vol 61 (5) ◽

pp. 402-415 ◽

Cited By ~ 9

Author(s):

Thorsten Meiser

Keyword(s):

Cognitive Processes ◽

Model Complexity ◽

Multinomial Model ◽

Stochastic Dependence ◽

New Model ◽

Multinomial Processing Tree ◽

Cognitive States ◽

The Family ◽

Tree Models ◽

Flexible Framework

Stochastic dependence among cognitive processes can be modeled in different ways, and the family of multinomial processing tree models provides a flexible framework for analyzing stochastic dependence among discrete cognitive states. This article presents a multinomial model of multidimensional source recognition that specifies stochastic dependence by a parameter for the joint retrieval of multiple source attributes together with parameters for stochastically independent retrieval. The new model is equivalent to a previous multinomial model of multidimensional source memory for a subset of the parameter space. An empirical application illustrates the advantages of the new multinomial model of joint source recognition. The new model allows for a direct comparison of joint source retrieval across conditions, it avoids statistical problems due to inflated confidence intervals and does not imply a conceptual imbalance between source dimensions. Model selection criteria that take model complexity into account corroborate the new model of joint source recognition.

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