The increased spontaneous fragility in the centromere regions of the chromosomes predisposes to abnormal cellular division due to abnormal interaction with the mitotic spindle, which leads to number chromosomal aberrations. The abnormal structure of the microtubule-associated proteins (MAPs) could also be among the internal factors, predisposing to these pathologies. On the other hand, the increased frequency of the spontaneous chromosomal fragility suggests abnormal structure of the respective chromosomal/chromatin components and thus an increased risk about gene mutations and structural chromosomal aberrations. The spontaneous chromosomal fragility was tested by light microscopy observation of metaphases from peripheral blood lymphocytes of 8 patients with polygene pathologies (neoplasms, neuro-degenerative diseases, and neuro-psychiatric disorders) and of 18 healthy controls. In the tested patients was established significantly higher frequency of the spontaneous chromosomal fragility, including in the centromere chromosomal regions, compared to the controls. Different inter-molecular interactions, which could prevent the pathology appearance, were investigated by appropriate experimental in vivo models. As a whole, in the most cases, the presented data showed significantly lower average titers of anti-ganglioside antibodies and gangliosides in the samples from myocardium and liver than in the samples from brain and pancreas. By taking into consideration that the myocardium and liver are known as the most enriched of GSH anatomic organs, the presented data could be explained with the literature messages of higher amounts of de novo-synthesized free GSH tri-peptide in the same two organs. Moreover, a possibility about the production of antibodies/immunoglobulins by non-lymphoid types of cells, tissues, and organs in appropriate conditions was shown. Because the so-produced antibodies are out of the germinative centers in the specialized lymphoid tissues and organs, the control of their functions by small ions and molecules as gangliosides is very important. In the current study, the attention was particularly directed to the iteractions with the participation of gangliosides, the reduced form of the tri-peptide glutathione (GSH), and the protein HACE1. Besides the tumor-suppressor activity of protein HACE1, also its neuroprotective influence was proven, namely by interactions with specific molecules.

Multifactor polygene diseases, experimental in vivo models, inter-molecular interactions, regulatory mechanisms

Fragile sites in the chromosomes have been determined as key breakpoints in the chromosomal rearrangements (Daniel 1986). Abnormal interaction with the fibers of the mitotic spindle, increasing in this way the risk of abnormal cellular division in the actively dividing cells and thus of number chromosomal aberrations, has been proposed. The abnormal expression of the microtubule-associated proteins (MAPs) is also an important reason for changes in the signal transduction in the affected cells (Arnold et al. 1991). Also, the increased frequency of the spontaneous chromosomal fragility proposes abnormal structure of the respective chromosomal/chromatin components and thus an increased risk of gene mutations and structural chromosomal aberrations. In this way, the spontaneous chromosomal fragility could be underlining many cases of multifactor pathologies such as malignancies, neuro-degenerative and cardio-vascular disorders, diabetes, etc. (Lukasa and Frins 2007).

As molecules, performing key protective and regeneration functions, preventing the development of malignancies and degenerative pathologies, have been determined gangliosides (Magistretti et al. 2019). They are acidic glycosphingolipids, known as regulators of many biological processes on cellular, tissue, organ, and organism levels, by influencing various inter-molecular interactions. Different changes in the gangliosides’ titers, types, composition, distribution, and proportions during the processes of development and aging, according to the healthy status, as well as in various pathologies, even in several cases with equal clinical symptoms, have been established (Zurita et al. 2001). These differences could vary between the separate cells, tissues, organs, and organisms. Furthermore, the existence of each ganglioside, besides in free form, also in various bind forms with different bio-molecules, depending on the respective functions in which it participates, should also be taken into consideration.

Among the main mechanisms that minimize the genomic instability has been proven the depletion of glutathione reduced form (GSH) (Hanot et al. 2012). GSH is known as a main immunomodulator, antioxidant, cardioprotector, neuroprotector, anti-neoplastic, and anti-aging substance, depending on the respective inter-molecular interactions in which this molecule participates (Meister 1983; Jahren-Hodje et al. 1997). Reduced intra-cellular GSH levels have been established in neurodegenerative diseases (Cacciatore et al. 2012).

One of the main proteins that regulates the distribution of cytoplasmic organelles in cell division is the protein HACE1 (Tang et al. 2011). This is an E3 protein ligase enzyme, which suppresses besides the malignancies also the neuro-inflammation and pathogenesis of neurodegenerative processes in experimental models of Parkinson’s and Alzheimer’s diseases (Jin et al. 2006). This function could be performed by regulating the activity of gene RAC1 and/or by degradation of protein Rac1, respectively. Additionally, a key role of the enzyme CUL7 E3-ubiquitin ligase in the proteolytic targeting of insulin receptor substrate-1 has been demonstrated (Zhang et al. 2014). Hence, this enzyme could be a critical mediator of the insulin/IGF1 signaling. Down-regulation and/or deletion of the HECT domain containing the enzyme E3 ubiquitin protein ligase 1 (the protein product of gene HACE1) is of key importance in the occurrence, invasion, metastatic process, and prognosis of many human malignancies (for instance, hepatocellular carcinoma) (Gao et al. 2016; Li et al. 2019). In this way, gene HACE1 as well as its protein product HACE1 have been determined as potential prognostic markers and therapeutic targets.

In this aspect, the main goal of the current study is to understand the inter-molecular interactions in different cells, tissues, and organs, which could prevent a multifactor disease appearance. For this aim, appropriate experimental in vivo models were developed. In the concrete case, the attention was particularly directed to the interactions with the participation of gangliosides, GSH, and/or HACE1 protein.

Peripheral blood from eight patients with neoplasms and neurodegenerative and neuro-psychiatric disorders (known to belong to the polygene multifactor pathologies), as well as from 18 healthy controls, was taken. The so-taken samples were ex vivo-incubated in 4 mL medium RPMI 1640 (Sigma-Aldrich) with previously added 0.8–1 mL fetal bovine serum (FBS) and 0.2 mL phyto-hemagglutinin at 37 °C in an incubator with 5% СО₂ and 95% air humidification for 68–70 hours. Subsequently, the further cell growth and proliferation were blocked by the addition of 0.2 mL Colchicin, and the samples were put back at 37 °C for 50 minutes. After careful shaking, they were centrifuged for 10 minutes at 1000 rpm. The supernatants were taken off, and 0.3 mL from the pellets were resuspended. 10 mL of warm hypotonic 0.555% KCl solution was added to each suspension, and the samples were put at 37 °C for 10 minutes. After centrifugation at 1000 rpm for 10 minutes, the supernatants were taken off. After resuspendation of 0.3 mL of the pellets, a 10 mL fixative mixture of methanol-glacial acetic acid (3:1) was added to each sample (pellet). After centrifugation at 1000 rpm for 10 minutes, this step was repeated 3–4 times by changing the fixative mixture. The prepared light microscopy slides were stained by the G-banding technique and observed by microscope Amplival with immersion to determine the frequency of fragility in the respective chromosomes from both patients and healthy controls. The differences were accepted as statistically significant at p < 0.05 and p < 0.01.

Total extracts from rodent brain, pancreas, myocardium, and liver were prepared by mechanical homogenization, followed by treatment with 10% trichloroacetic acid (Cl₃CCOOH) and a 0.48 M solution of K₃PO₄. The samples were subsequently centrifuged at 3000 × for 10 minutes. Each sample was separated in two parts: controls, containing the full composition of molecules of the respective anatomic organ, as well as samples, containing molecules with affinity to GSH, received by passing through GSH-agarose columns (Cuattrecasas 1970).

All prepared samples from the four rodent anatomic organs were subjected to an enzyme-linked immuno-sorbent assay (ELISA) to assess the average titers of anti-ganglioside antibodies and of gangliosides in them. In all cases, the received titers were expressed as mean ± standard deviation (SD). The differences were considered statistically significant at p < 0.05 and p < 0.01.

Fixed light microscopy slides from the brain, pancreas, myocardium, and liver with rodent (rat) origin, stained by the hematoxillin/eosin (H/E) technique, were prepared. For this goal, slices from the four anatomic organs were fixed in 10% neutral phosphate buffered formalin and subsequently included in paraffin. The prepared paraffin slices were dehydrated in an ethanol series and subjected to H/E staining. The stained and dried slides were observed by an inverted light microscope, Televal, supplied by a mega-pixel CCD camera.

The normal presence and expression of the tested tumor suppressor gene HACE1 in normal/non-malignant mature cells in the body were studied by standard polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR), respectively. Subsequent electrophoresis in 1% agarose gel was performed. Appropriate primers, complementary to the cellular genome, with sizes 750–2500 base pairs (bp), were applied. Cellular DNA (total genomic material) was isolated from mature epithelial cells of the tail skin from two inbred lines of adult mice: Wild Type (WT) Dcn1+/Dcn1+ homozygotes and partially Knocked Down (KD) Dcn1+/Dcn1- heterozygotes, respectively, but also from experimental mice of line Balb/c. A single-strand conformational polymorphism (SSCP) assay was applied, taking into consideration the protocol developed by Dong and Zhu (2000).

A significantly higher frequency of the spontaneous chromosomal fragility in the patients compared with the controls was established (Fig. 1). The most often affected was chromosome #11 (p < 0.01), followed by chromosomes #1 (p < 0.05) and #17 (p < 0.01).

Figure 1. 

Frequency of the spontaneous chromosomal fragility in patients with polygene multifactor disorders and in healthy controls.

The higher frequency of the spontaneous chromosomal fragility in the patients compared with the healthy controls was established in human white peripheral blood cells. Experimental in vivo models of several types of tissues and organs were developed to understand the inter-molecular interactions, which could prevent multifactor disease development.

In most cases, increased average titers of both anti-ganglioside antibodies and gangliosides in the samples from rodent brain and rodent pancreas containing molecules with affinity to GSH were noted than in the control samples (Fig. 2A, B, D, E, G, H). Only at dilution 1:400 of the rodent brain total extract was a statistically significant difference between the average titers of the anti-ganglioside antibodies (Fig. 2A) and of the gangliosides (Fig. 2B) between the controls and the samples, containing molecules with affinity to GSH. Approximately equal average titers of the gangliosides and the anti-ganglioside antibodies in the total pancreas extract and in the sample of the same organ, containing molecules with affinity to GSH, were assessed at dilution 1:100 (Fig. 2D). At dilution 1:40, the average titers of the gangliosides in the total lysate of pancreas and in the sample containing molecules with affinity to GSH were also approximately equal (Fig. 2D). At the same dilution of the total pancreas extract, a significantly higher average titer of the anti-ganglioside antibodies was noted compared with the sample of the same organ lysate, containing molecules with affinity to GSH. In most of the cases, in the samples of rodent pancreas, the average titers of the anti-ganglioside antibodies were significantly higher (Fig. 2D) compared with the average titers of the gangliosides (Fig. 2E). Only at dilution 1:200 of the pancreas lysate were noted higher average titers of the anti-ganglioside antibodies and of the gangliosides in the total organ lysate compared with the sample of the same organ, containing molecules with affinity to GSH (Fig. 2D, E). This difference was statistically significant about the average titers of gangliosides (Fig. 2E) and not statistically significant about the average titers of the anti-ganglioside antibodies (Fig. 2D). The opposite tendency was assessed in the samples of the myocardium extract (Fig. 2G, H). Only at dilution 1:40 was assessed a significantly higher average titer of the anti-ganglioside antibodies in the sample, containing molecules with affinity to GSH compared to the total extract of the same anatomic organ (Fig. 2G). At all other dilutions of the same samples, the opposite tendency was noted – significantly higher average titers of the anti-ganglioside antibodies in the total myocardium extract compared to the samples containing molecules with affinity to GSH.

Figure 2. 

Samples showing some general biochemical and morphological characteristics between rodent brain, pancreas, and myocardium: A, B, D, E, G, H are average titers in total organ lysates and in samples containing selected molecules possessing affinity to GSH by passing through an agarose-GSH column. A of the anti-ganglisodie antibodies in samples of rodent brain; B of the gangliosides in samples of rodent brain; D of the anti-ganglisodie antibodies in samples of rodent pancreas; E of the gangliosides in samples of rodent pancreas; G of the anti-ganglioside antibodies in samples of rodent myocardium; H of the ganglisodies in samples of rodent myocardium. On the abscise are shown the dilutions of the respective biological samples in µM/g tissue material, and on the ordinate are the values of the optical density (OD); C, F, I, L are fixed histological preparations, stained by H/E; C of rat brain; F of rat pancreas; I of rat myocardium; and L of rat liver.

The highest average titers of gangliosides at dilutions 1:100 and 1:200 of all samples of rodent brain extract compared with the data at dilutions 1:40 and 1:400 were observed (Fig. 2B). These data could be explained by the presence of many other non-specific molecules at the lowest dilution (1:40), but an insufficient amount of gangliosides at the highest dilution (1:400) in equal volumes of biological material. Due to the same reason are probably the established highest average titers of the anti-ganglioside antibodies at dilutions 1:100 and 1:200 compared to dilutions 1:40 and 1:400 of the total brain extract (Fig. 2A). This explanation could also be given about the noted highest average titers of the gangliosides at dilutions 1:100 and 1:200 of the total rodent liver extract compared to the results at dilutions 1:40 and 1:400 of the total lysate of the same organ (Fig. 2K). In most of the cases, significantly higher average titers of the gangliosides were observed in the total myocardium extract compared with the sample of the same organ, containing molecules with affinity to GSH (Fig. 2H). Only at dilution 1:400 were established equal average titers of the gangliosides in the two samples of the myocardium extract. At dilutions 1:40 and 1:100 of the total liver extract, significantly higher average titers of the anti-ganglioside antibodies compared to the sample containing molecules with affinity to GSH (Fig. 2J). The opposite tendency was established at dilutions 1:200 and 1:400 of the samples from the same organ lysate – significantly higher average titers of the anti-ganglioside antibodies in the sample, containing molecules with affinity to GSH compared to the average titers of these antibodies in the total liver lysate. In most of the cases, significantly higher average titers of gangliosides were assessed in the total liver extract compared to the sample of the same organ, containing molecules with affinity to GSH, with the exclusion of the dilution 1:40, where the opposite tendency was observed (Fig. 2K). As a whole, significantly lower average titers of the anti-ganglioside antibodies and of the gangliosides were assessed in the samples from rodent myocardium (Fig. 2G, H) and rodent liver (Fig. 2J, K) compared with the same samples from rodent brain (Fig. 2A, B) and rodent pancreas (Fig. 2D, E). Morphological features in each of the four tested anatomic organs were assessed by observation of light microscopy slides (Fig. 2C, F, I, L).

The normal presence and expression of the tumor suppressor gene HACE1 in the normal/non-malignant cells in the body were confirmed by performing standard PCR and RT-PCR, respectively. Cellular DNA (genome material) was isolated from mature cells of the tail skin of adult experimental mice from the two inbred lines described above (Fig. 3A), but also from Balb/c mice (Fig. 3B). Specific primers, complementary to the cellular DNA genome, were applied. Besides in the DNA profiles of the standard PCR between the two inbred lines of experimental mice, varieties in the DNA profiles of the RT-PCR between them were also noted. These features suggested differences besides in the tested genes in the cellular DNA genome of the experimental mice from the two inbred lines and also in the respective mRNA transcripts of each one of the two genes of interest – the tumor suppressor gene HACE1 and the opposite oncogene Dcn1, respectively.

Figure 3. 

Agarose gel electrophoresis of samples from mature cells of adult experimental mice: A: presence (lanes 2 and 4) and expression (lanes 3 and 5) of tumor suppressor gene HACE-1 in each one of the two inbred lines of experimental mice: lanes 2 and 3 – in WT Dcn1+/Dcn1+ homozygotes; lanes 4 and 5 – in partially KD Dcn1+/Dcn1- heterozygotes, assessed by standard PCR (lanes 2 and 4) and RT-PCR (lanes 3 and 5). Specific primers, complementary to the cellular DNA genome, were applied. Lane 1: specific molecular marker (M) with size 750–2500 bp. B: in Balb/c experimental mice: lanes 2–6: normal presence (lane 2–4) and expression (lanes 5 and 6) of oncogene Dcn-1; normal presence (lanes 8–10) and expression (lanes 11 and 12) of the opposite tumor suppressor gene HACE1 in mature epithelial cells from the skin of an adult animal, assessed by standard PCR (lanes 2–4 and 8–10) and RT-PCR (lanes 5, 6, 11, and 12). Specific primers, complementary to the cellular DNA genome, are applied. Lane 1: specific molecular marker (M) with size 750–2500 bp.

Significantly lower amounts (Fig. 3A, lane 2) and expression levels (Fig. 3A, lane 4) of the tumor suppressor gene HACE1 were noted in the homozygotes on the opposite oncogene Dcn1 compared to its presence (Fig. 3A, lane 3) and expression (Fig. 3A, lane 5) in the heterozygotes Dcn1 gene. These variations were also observed in comparison of each one of both inbred lines (Fig. 3A) with the data from the experimental mice of line Balb/c (Fig. 3B). In the last was assessed normal presence (Fig. 3B, lanes 8–10) and expression (Fig. 3B, lines 11 and 12) besides of the tumor suppressor gene HACE1, also normal presence (Fig. 3B, lanes 2–4) and expression (Fig. 3B, lanes 5 and 6) of the opposite oncogene Dcn1.

The centromere regions have been proven as places where are located many genes, coding long non-coding RNAs (lncRNAs) as ribosomal RNAs (rRNAs), transport/transfer RNAs (tRNAs), etc. Centromeres have also been characterized as the main structures of the cellular nucleoli, and in the last years they have been proven among the main targets about epigenetic influence and reparation (Dong and Zhu 2000; Bersaglieri and Santoro 2019). The noted features could suggest the role of centromere regions in the chromosomes (of the cellular nucleoli, respectively) as appropriate targets about gene diagnostics as well as about genetic and epigenetic regulation. Transposon-rich centromere regions have been found to be favored by RNAi machinery, which silences the transposons by suppression of meiotic cross-overs at centromeres and/or by their targeting (Talbert and Henikoff 2022). Thus, the centromere regions could reveal new genetic and epigenetic properties (Mattick et al. 2023). The role of the cytoskeleton components in the neuro-transmission processes in the brain has been proven (Butner and Kirschner 1991; Nguyen-Ngoc et al. 2007; Zhang et al. 2010). In the human chromosome #1, which is shown among the most frequently affected of spontaneous fragility, is localized the gene coding trans-active DNA-binding protein-43 (TAR DNA-binding protein 43/TDP-43) (https://www.ncbi.nlm.nih.gov/gene/23435). The abnormal variations of this gene have been found to form aggregates in the cells of the brain and spinal cord, disrupting the normal functions in the neural cell. Furthermore, these aggregates have also been determined as a common pathological feature of amyotrophic lateral sclerosis (ALS/Lou Gerig’s disease), fronto-temporal dementia (FTD) (Tran and Lee 2022), fronto-temporal lobar degeneration (FTLD), as well as other neuro-degenerative disorders, including some forms of encephalopathy, Alzheimer’s, and Parkinson’s diseases (Lee et al. 2018).

The neuro-protective role of the gangliosides in the neural cell nucleus has been indicated, including after the ischemic brain injury by inhibition of the following neuronal autophagy (Sasaki et al. 2019). Furthermore, the role of gangliosides on important signaling pathways has been proven, which has delineated these molecules as appropriate targets for anti-neoplastic therapeutic directions (Jaumot et al. 1994). In this way, the presented results suggest different activities of gangliosides by their participation in various inter-molecular interactions. Additionally, the received data were in agreement with literature findings about the inter-molecular interactions with the participation of gangliosides in their role as neuroprotectors. In this context, mutations in the gene coding the muscle-specific immunoglobulin domain-containing protein myopalladin (MYPN) have been associated with cardiomyopathy development (Filomena et al. 2021). Recently, protein MYPN, as well as other members of the family to which it belongs, have been found to bind to the filamentous actin (F-actin), thus preventing actin depolymerization (Cutillo et al. 2020). MYPN protein has also been established as capable of binding to myocardin-related transcription factors A and B (MRTF-A and MRTF-B). In this way, this protein could provide a shuttle between the cytosol and the nucleus in response to alterations in actin dynamics. Additionally, MYPN binds to various z-line proteins in the myocardium, thus providing efficient signal production and transmission (Bang 2017). The observed features on light microscopy slides from each one of the four tested anatomic organs were in agreement with the literature data about the importance of the different methods for assay in a better understanding of the metabolic processes (Cabrera et al. 1998). On the other hand, the anti-ganglioside antibodies have been proven to target the immune reactions against neuronal cells and to neutralize their complement inhibitory activity (Xu et al. 2010; Li et al. 2021). In this way, our results were in confirmation of the literature data about various mechanisms of the anti-malignant functions of the different gangliosides. These mechanisms could underline the activated apoptosis of the injured cells, as well as anti-neoplastic actions expressed in enhanced activity of tumor suppressor genes and/or of their protein products. The current results were also in agreement with the literature findings about the possibility of expression of genes, coding immunoglobulins, and/or their components/domains by non-lymphoid types of cells, tissues, and organs (Bebbington 1991; Deyev et al. 1993). The shown property of non-immune cells to produce immune molecules directs attention to the development of novel diagnostic, prophylactic, and therapeutic approaches. One of the explanations about the production of immune molecules by non-immune cells is related to the proven in the scientific literature possibility of sub-populations of immature stem-like cells to acquire initial lymphoid and myeloid properties (McDonnald et al. 1997). According to another hypothesis, functions of immunoglobulins could perform molecules, which act as enzymes in the pancreas (Takeda et al. 1989) and/or as neurofillaments in the neurons (Otey et al. 2009). These properties could be among the underlining components in regulatory mechanisms, by which different tissues and organs communicate with each other. Because the so-produced immune molecules and/or their components are out of the germinative centers in the specialized lymphoid tissues and organs, the control of their production and functions by appropriate inter-molecular interactions is of key importance. In this relation, we investigated the interactions and influences of small ions and molecules as gangliosides. In this way could be proposed new methods about reparation of gene mutations leading to neurodegenerative pathologies, by transfer of appropriate genes/DNA fragments or by application of appropriate RNAs against the respective abnormal genes or genomic variations. The current results showed significantly lower average titers of the anti-ganglioside antibodies and of the gangliosides in the samples from myocardium and liver compared to the samples from brain and pancreas. By taking into consideration that the myocardium and liver are known as the most enriches of GSH anatomic organs, these data could be explained with the proven in the literature higher amounts of de novo-synthesized free GSH tri-peptide in the same two organs compared with brain and pancreas, in which probably the molecule participates in various inter-molecular interactions (Lu 2013). The presented results supported the literature data about the link and protective role of gangliosides and GSH against neuro-degenerative processes and diabetes (Jahren-Hodje et al. 1997). In the current study, the determination of the variations in the tested biochemical parameters in the separate tested anatomic organs was combined with the assessment of the specific structural features by light microscopy observation. Similar combinations of physiological systems approach and mathematical model development have been applied to a better understanding of the inter-molecular interactions, metabolic processes, and the mechanisms of their regulation (Cabrera et al. 1998).

According to the current data, besides the observed differences in the normal presence and expression of the tumor suppressor gene HACE1 between the two inbred mice lines, differences were also noted in the normal presence and expression of its opposite oncogene, Dcn1. Oncogene Dcn1, which in humans is located in chromosome 12, codes the enzyme NEDD8 ligase (https://www.ncbi.nlm.nih.gov/gene/850819) (Zhang et al. 2017). This ligase has been determined as a novel target for the treatment of prostate cancer, in which it is highly expressed. A possibility about neuro-protective functions of tumor suppressor proteins (on the level of gene products) together with the preservation of their anti-malignant actions was proven. Tumor suppressor gene HACE1, which in humans is localized in the long arm of chromosome 6 (https://www.ncbi.nlm.nih.gov/books/NBK470040), codes protein-kinase, which has been proven as a target for degradation of key proteins, responsible for malignancy and metastasis processes (Zhang et al. 2007). The relationships between DNA replication, chromatin, and proteolysis have been confirmed by proved cullin-RING E3-ubiquitin ligases assembled on the CUL4 platform (Anzai et al. 2020). Besides against neoplasms, HACE1 protein has shown protective influence against many neurodegenerative pathologies, as well as cardiac and infectious diseases (Zhang et al. 2014; Zang et al. 2024). Similarities in maturation marker genes between myocardium of mouse, rat, and human origin have recently been shown (Okada et al. 2022).

In the current research paper were performed initial studies on genes and molecules, which abnormalities could lead to malignancies, neuro-degenerative, and neuro-psychiatric disorders. Various inter-molecular interactions, underlining the regulatory mechanisms, that could prevent the pathological process, were investigated by appropriate experimental in vivo models. The assessed varieties in the average titers of the gangliosides and of the anti-ganglioside antibodies could be related to the differences in the structure and functions of the separate cells, tissues, and organs. As a whole, in most of the cases were observed significantly lower average titers of the anti-ganglioside antibodies and of the gangliosides in the samples from myocardium and liver than in the samples from brain and pancreas. By taking into consideration that the myocardium and liver are known as the most enriched of GSH anatomic organs, the current results could be explained with the established, according to many literature messages, higher amounts of de novo-synthesized free GSH tri-peptide in the same two organs. Additionally, a possibility about the production of antibodies/immunoglobulins by non-lymphoid types of cells, tissues, and organs in appropriate conditions was proposed. Because the so-produced antibodies are out of the germinative centers in the specialized lymphoid tissues and organs, the control of their functions by small ions and molecules as gangliosides is of key importance. One of the explanations is that gangliosides participate in various inter-molecular interactions. In the concrete study were taken into consideration the interactions with participation of gangliosides, GSH tri-peptide, and protein HACE1. Despite the importance of protein HACE-1 as a tumor suppressor, its role to prevent the development of many neurodegenerative processes by appropriate inter-molecular interactions was confirmed.

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statements

Clinical trials: All human rights are taken in consideration

The authors declared that no experiments on humans or human tissues were performed for the present study.

The authors declared that no informed consent was obtained from the humans, donors or donors’ representatives participating in the study.

Experiments on animals: All rights of the work with experimental animals are taken in consideration according to Helsinki rules

The authors declared that no commercially available immortalised human and animal cell lines were used in the present study.

Funding

No funding was reported.

Author contributions

I.S. – main idea and research; V.K. – statistical assay; V.N. – light microscopy photos; D.D. – biological materials (sera and blood samples) from human patients with neurodegenerative and neuro-psychiatric disorders; D.M. – relation with the tested patients with neurodegenerative and neuro-psychiatric disorders; D.D. –providing of siurces, necessary with experiments with the tested experimental models; Tz.M. – providing the tested experimental models.

Author ORCIDs

Dimitrina Dimitrova-Dikanarova  https://orcid.org/0000-0001-5399-6954

Data availability

All of the data that support the findings of this study are available in the main text.