Amended on 14 June 2019

I have searched Adenosine A1 receptor, Actin, Exosome-related protein, Microtubule protein of Tubulin alpha and beta, and Guanylyl cyclase in vain among my human protein database (please see files; HepG2 fucoidan, JMBT Alopecia and The Fascio effect). This seems to be due to famous Species differences. Then, Biochemical studies using Rat are contributing to Veterinary medicine. Aquaporin, AChR, GABA(A)-R, and Usher syndrome protein are present in Humans. Human serum biotinidase seems to differenciate Human hormones and other Animal Hormones (please see file; Enk serBIN). Therefore, I am now studying Human Receptors in general.

We have already found that enzyme kinetics (Biotinidase kinetics) are different between right cerebrum and left cerebrum in the LEW Rat (please see Fig. 1 of the file; J Chrom B Rat BIN LIP Km). Therefore, study of Müller glia/Müller cells should be performed by using both Müller cells derived from Right eye and from Left eye. Enzyme study should be performed by utilizing sensitive and reliable HPLC-photometric and/or HPLC-fluorimetric method. Please do not utilize un-reliable and non-quantitative neither ELISA method, Immunological method (using IgG). nor Avidin (binding protein to biotin/vitamin H and Lipoic acid/Thioctic acid) binding method.

Healed hepatocyte HepG2 (cultured with fucoidan at 0.102 mg/mL for 3 days) has Adenylyl cyclase-associated protein 1/CAP-1 at 0.25 μg/mg of cell protein (please see again; HepG2 Fucoidin). This protein may be working as a membrane protein transporter (protein localization to plasma membrane). Therefore, Fucoidan can increase the ratio of Membrane components. Increase of Membrane components in Fucoidan-treated HepG2 cells is also helped by Lamp-1/Lysosome-associated membrane glycoprotein 1 at 0.21, MICAL-like protein 2/Molecule interacting with CasL-like 2 at 1.1, Protein transport protein Sec 61 subunit alpha isoform 1 at 0.6, Secretory carrier-associated membrane protein 1 at 0.31, Sorting nexin-6 at 1.1, and TNF receptor-associated factor 2/Tumor necrosis factor type 1 receptor-associated protein 2 at 1.05 μg/mg of cell protein, respectively.

I am now considering that cAMP and/or cGMP is only a second messenger of Rat, but they are not the second messenger of Humans. Rhodopsin-like G-protein coupled receptors seem to utilize phosphatidylinositol-calcium (PI/Ca) and/or other Ions as second messengers in Humans.

Aquaporin, AChR, GABA(A)-R, and Usher syndrome protein are present in Humans. Therefore, I have determined the amounts of these proteins utilizing PI/Ca or Ion channel system, and the correlation to the up-regulating virus are searched by using Data Mining method. Data Mining is performed on reliable and quantitative Human protein database made by PDMD (Protein-Direct-Microsequencing-Deciphering) method.

It is found that proteins utilizing PI/Ca or Ion channel system are up-regulated by Retrovirus and Coronavirus.

I am grateful to Dr. Kristina Forsch (Max Zeller & Söhne AG, Seeblickstrasse 4, Romanshorn, Switzerland) for leading me to the role of Retrovirus such as HIV-1 in the gene/protein expression. Human hepatocellular carcinoma (HCC) is caused by the co-infection of HIV-1 (Human-specific Retrovirus) and HCV (Human-specific Flavivirus (Hepacivirus)) (please see file again; HepG2 Fucoidin). Further, Lethal HCC is caused by the co-infection of HIV-1, HCV, and GB virus C/GBV-HGV/GBV-C/HGV (Pig-specific Flavivirus (Pegivirus)) and/or Hog cholera virus/Classical swine fever virus/CSFV (Pig-specific Flavivirus (Pestivirus)) (my unpublished observation).

Thank you for your feedback. I am currently trying to perform a cAMP accumulation assay (Lance by Perkin Elmer) in the Müller cells. Müller cells express, according to literature, all three adenosine receptors. I get a good concentration dependence after treatment with forskolin, but as soon as I incubate forskolin (1 μM) with a concentration series of CCPA (50 nM - 0.195 nM) I do not see any concentration dependence at all. According to the literature, CCPA is a very selective A1 receptor agonist. Do you have any idea why it does not work?