On 2014 Apr 26, Salvatore Chirumbolo commented:

This interesting paper by Marzotto M et al., evaluated gene expression after 24 hrs incubation with hydroalcoholic/water extracts of Gelsemium sempervirens plant. The authors used human neuroblastoma cell lines SH-SY5Y and also IMR32 cell line, to focus on previously published data on mouse behavioural tests. After an oligonucleotide microarray, the authors selected less than 10 genes for RT-PCR. However, using human neuronal cell line to elucidate results obtained with mouse models appears quite disputable. The Authors referred this study to the previously reported behavioural evidence on animals, when stated (Discussion pag 15) “In previous recent trials, Gelsemium s. showed anxiolytic-like effects in mouse emotional response models and appeared to work even at the high dilutions 9c and 30c [26,27]” ....and moreover “In general, the prevalence of down-regulation seems to indicate a tendency to reduce cell excitability, especially because several of the genes in question belong to surface receptors involved in GPCR signaling and calcium homeostasis. Moreover, this first microarray screening of the effects of Gelsemium s. on neurocytes revealed a significant down-regulation of genes for inflammatory response, olfactory transduction and neuron differentiation” (Discussion pag. 16). This may raise criticism, as the authors studied SH-SY5Y and IMR32 human neuroblastoma cell line, not mouse derived cell lines. Yet, several genes the Authors highlighted do not have homologues in mice. Some examples of this are reported as follows. The gene baculoviral IAP repeat containing 8 (BIRC 8) has no homologues in mice (only orthologue genes in Pan troglodytes), olfactory gene OR4X1 is not expressed (absent) in mouse, gene C1ORF167 has a non characterized orthologue gene LOC102634746 in mouse, certainly not matching the research purpose to relate olfactory gene to the behavioural test. The search for an involvement of neural genes related to anxiety/depression or mood disorders is biased by the expression of human genes having no orthologues/homologues in mice, where the authors reported evidence about Gelsemium action on behavioural tests in animal anxiety models. Genes such as EN2 (engrailed homeobox 2), GALR2 (galanin receptor 2), GPR25 (G-protein coupled receptor 25), KLKBL14, erroneously indicated by the authors as Klkbl14, namely PRSS54 (protease serine 54), tachykinin 4 or TAC4 have orthologues both in human and mouse, while OR5C1 (olfactory receptor 5C1) is an exclusive Homo sapiens gene (mouse has an olfactory receptor 368 gene on chromosome 2). The authors tested Gelsemium hydroalcoholic preparations without a Limulus test to prevent bias from contamination by LPS or bacteria: this may generate bias. Curiously, genes affected by Gelsemium 2c, such as DDl1, are bacterial genes, notoriously absent/non expressed in Homo sapiens: probably is a misprint (delta-like 1 (DLL1) instead of D-alanine-D-alanine ligase (DDl1)). “The genes investigated by quantitative PCR generally confirmed the changes obtained by microarray assay. DDl1, EN2, GALR2, GPR25, OR5C1, Klkbl4 and TAC4 genes were downregulated in Gelsemium s. 2c samples compared to Control 2c in the three replicated experiments” (Results pag. 7). DDl1 is a bacterial gene: maybe the authors would indicate DLL1 (delta-like 1 gene)?

On 2014 Apr 26, Salvatore Chirumbolo commented:

This interesting paper by Marzotto M et al., evaluated gene expression after 24 hrs incubation with hydroalcoholic/water extracts of Gelsemium sempervirens plant. The authors used human neuroblastoma cell lines SH-SY5Y and also IMR32 cell line, to focus on previously published data on mouse behavioural tests. After an oligonucleotide microarray, the authors selected less than 10 genes for RT-PCR. However, using human neuronal cell line to elucidate results obtained with mouse models appears quite disputable. The Authors referred this study to the previously reported behavioural evidence on animals, when stated (Discussion pag 15) “In previous recent trials, Gelsemium s. showed anxiolytic-like effects in mouse emotional response models and appeared to work even at the high dilutions 9c and 30c [26,27]” ....and moreover “In general, the prevalence of down-regulation seems to indicate a tendency to reduce cell excitability, especially because several of the genes in question belong to surface receptors involved in GPCR signaling and calcium homeostasis. Moreover, this first microarray screening of the effects of Gelsemium s. on neurocytes revealed a significant down-regulation of genes for inflammatory response, olfactory transduction and neuron differentiation” (Discussion pag. 16). This may raise criticism, as the authors studied SH-SY5Y and IMR32 human neuroblastoma cell line, not mouse derived cell lines. Yet, several genes the Authors highlighted do not have homologues in mice. Some examples of this are reported as follows. The gene baculoviral IAP repeat containing 8 (BIRC 8) has no homologues in mice (only orthologue genes in Pan troglodytes), olfactory gene OR4X1 is not expressed (absent) in mouse, gene C1ORF167 has a non characterized orthologue gene LOC102634746 in mouse, certainly not matching the research purpose to relate olfactory gene to the behavioural test. The search for an involvement of neural genes related to anxiety/depression or mood disorders is biased by the expression of human genes having no orthologues/homologues in mice, where the authors reported evidence about Gelsemium action on behavioural tests in animal anxiety models. Genes such as EN2 (engrailed homeobox 2), GALR2 (galanin receptor 2), GPR25 (G-protein coupled receptor 25), KLKBL14, erroneously indicated by the authors as Klkbl14, namely PRSS54 (protease serine 54), tachykinin 4 or TAC4 have orthologues both in human and mouse, while OR5C1 (olfactory receptor 5C1) is an exclusive Homo sapiens gene (mouse has an olfactory receptor 368 gene on chromosome 2). The authors tested Gelsemium hydroalcoholic preparations without a Limulus test to prevent bias from contamination by LPS or bacteria: this may generate bias. Curiously, genes affected by Gelsemium 2c, such as DDl1, are bacterial genes, notoriously absent/non expressed in Homo sapiens: probably is a misprint (delta-like 1 (DLL1) instead of D-alanine-D-alanine ligase (DDl1)). “The genes investigated by quantitative PCR generally confirmed the changes obtained by microarray assay. DDl1, EN2, GALR2, GPR25, OR5C1, Klkbl4 and TAC4 genes were downregulated in Gelsemium s. 2c samples compared to Control 2c in the three replicated experiments” (Results pag. 7). DDl1 is a bacterial gene: maybe the authors would indicate DLL1 (delta-like 1 gene)?