**3.3 Anaerobic synergistic preparations containing LSSM for support of human protective systems**

Due to high distribution in organism, oxidative stress (as the power destructive factor initiating diseases) needs the constant presence of the power protective antioxidant systems [28, 29]. Some therapeutic proteins regulating cellular consumption of oxygen can be involved into development of tumor and other side pathologies in organism. We isolated system anaerobic (without oxidases initiating of oxygen and peroxide radicals) preparations of acidic/anionic and alkaline/cationic LSSM from cultures of symbiotic (probiotic) industrial strains of human bifidobacteria and lactobacilli as consortia that were successfully applied. Such preparations devoid the ability to induce destructive oxidative stress (crosslinking and inactivation of therapeutic proteins, etc.) in respect to surrounding infrastructures.

The used synthetic GC in our work were characterized with antioxidant properties in respect to LS as carriers of GC (prolongation of chemiluminescence of protective complexes was observed). Similar resulting protection of LSSM was also registered in the presence of neutral and cationic bifidobacterial and lactobacillar cultural exopolymeric compounds (EPC) of nonprotein origin (as observed on the blot after IEF-PAG). Acidic and alkaline anaerobic LS of bifidobacteria and lactobacilli revealed the following general antipathogenic actions: (a) own and overlapped/synergistic; (b) toward communicative bodies of microbial massifs and biofilms of the potentially pathogenic yeast-like fungi and Gram-positive bacteria. All four types of preparations of LSSM used were characterized by own mechanisms of antimicrobial actions in comparison to action of other antimicrobial systems (antibiotics, bacteriocins, phytolectins, subisotype products of isotypes С4В and С4А of the human complement component C4) [30, 31]. LS from human probiotic bacterial cultures revealed the ability to act as cascades in such reactions as initiation/changing or switching recognition of GC of different types (imitators of mannans, mucins, components of bacterial walls, Forssman antigens, Tn, blood group substance A) using the same original pool of lectin forms of taken multistrain probiotic. The presence of cations Ru2+ (ingredient of SYPRO involved in photosensibilization) strongly increased discreteness and number of forms of acidic lectins—potential carriers and deliveries of GC. Stability of obtained mosaic asymmetric landscape pictures of the systems LSSM-GC as multistrain probioticdepending and multistrain probiotic-supporting biotope balance of recognition and reversible retaining/depositing of GC (therapeutics, biomarkers, others) was observed. Combinations of anaerobic LS-containing proteins revealed themselves in respect to yeast-like and Gram-positive pathogenic targets as more selective in the choice of the adequate regional territory of massif of pathogen and limitation of early and late time (depending on localization of targeted region of communicative

body of pathogen) for the mostly effective visible actions of LS, obtaining uniform pure landscapes of LSSM action on massif of *Candida albicans* (the absence or minimization of LSSM-resistant residual fungal colonies in the interacting intestine system "LS of human intestinal bifidobacteria and lactobacilli—human intestinal *C. albicans*"). Antimicrobial activities of LSSM and phytolectins (phytohemagglutinin from the kidney bean) could be realized not only directly but also through the influence (together with synthetic mannans and mucins) in respect to macrophage migration as well as through inducing production of cytokines by stimulated blood lymphocytes (on the example of tumor necrosis factor-α). Results indicate prospects of anaerobic LSSM as assistant ingredients of the possible drug forms.

#### **3.4 Synbiotic minibioreactor using LSSM for screening GC**

During the last time, synbiotics and symbiotics (as synergistic sum of probiotics and prebiotics) are of increased investigator interest due to their antimicrobial and other useful reactions [1, 2]. In this respect LSSM represent new class of antipathogenic proteins (possessing extended potential of application) which recognize different GC. LSSM represent multifunctional potential of relatively highly molecular mass polymeric metabolites of cultures of the human microbiota (microbiocenoses) and consortia (also multistrain probiotics) of human indigenous microorganisms. LSSM cofunction together with artificial and biologically active natural GC [32].

According to own results, we proposed suitable laboratory minisystem for screening prebiotic and therapeutical GP using LSSM and sterile heparinized insulin syringes of 1 ml volume. The following results were obtained: (1) LSSM-containing fraction stimulated production of both the whole and adhesive mass of bifidobacteria, (2) LiCl (15 mM and higher) increased dose depending on the number of adhesive colonies, (3) bifidobacterial LSSM (within pI 4–4.5) were characterized with strong affinity to anionic synthetic GC (possessing exposed residues of sulfated galactosides or, in a less extent of affinity, exposed residues of mannose-6-phosphates), and (4) sulfated glycosaminoglycans together with cations Li+ and LSSM as potential carriers of Li+ participated in functioning bioreactor imitating synbiotope (multiplication of bifidobacterial colonies and their survival were observed).

Proposed synbiotic system is perspective for screening prebiotic GC (as it is known for prebiotic derivatives of chitin, chitosan, fucoidan, and glycopeptides [33, 34]). **Table 1** includes potential prebiotic sources such as chitin and α-L-fucan which react to LSSM (LSSM may serve carriers of metabiotic GC; the list of GC can be unlimitedly extended).

#### **3.5 Membrane technological prospects of LSSM**

Progress in membrane and solid-phased technologies using LS-GC interactions includes the potential of their application in microassays, biochips (membrane bound glycoarrays or lectin arrays), and biosensors [35, 36].

The use of affine pore hydrophobic (uncharged) membranes predictably covered with mosaics of multifunctional sets of LSSM (additional significant purification of LSSM on membranes is reached) allows prolonged storing LSSM without decreasing samples in activities. The following prospects of LSSM-GC combinations may be of practical interest: (a) antifungal covers of prolonged action in combination with antibiotics and physical factors of stress [radiation (ultraviolet and ultrasonic), light (biorhythm "day-night"), temperature, pH, oxygen, season changes (also biorhythmic), others] and (b) chemiluminescent systems cofunctioning in regime of real time for medical and industrial biotechnology and

**83**

*Metabolite Multiprobiotic Formulas for Microbial Health DOI: http://dx.doi.org/10.5772/intechopen.86449*

understandable, and reliable results.

bionanotechnologies [our results include the following coupled systems: "low acidic LSSM, low acidic oxidoreductases of lactobacilli"; "alkaline bifidobacterial LS, alkaline bifidobacterial exopolymeric compounds"; "neutral lactobacillar/bifidobacterial LS, neutral lactobacillar/bifidobacterial biosurfactants"; "LS, strongly acidic

Membrane technologies using separated proteins, oligopeptides, and their complexes (especially) together with intrinsic or exogenic (SYPRO dye) fluorescence registered in live bioimagination are especially sensitive and perspective (protein band discreteness using fluorescence technique was better compared to the chemiluminescence technique). The latter allows identification stabile boundaries of the whole protein massifs for further establishment of LSSM and other biologically and physiologically active components among protein mosaics. Bioluminescence (fluorescent technique in combination with chemiluminescence technique) in optimal (depending on the object and the goal of study) conditions allows express-ranging cultural fluid groups of proteins and LS according to molecular mass (for additional standardization and typing of strains), evaluation of interstrain synergism and contribution of protease and oxidoreductase systems of mono- and multistrain probiotics (symbiotics) and other type consortia, and identification of mosaics of complexes containing fluorochromes in extended interval of pI/pH (complexes and cell wall fragments as carriers of visible energy which is ready for energetic exchange with surrounding infrastructures as well as for monitoring directed supramolecular assembling and their reorganization). Aforementioned data presented develop other possible important prognostic approaches and proposals. The choice of wished prognostic (LSSM-selected-type and GC-type interactions)-directed events in biotope can be determined and regulated by involving GC types needed (panels of natural and artificial GC used in biotope; extending the list of GC indicated in **Table 1** is possible). The use of artificial polymeric GC with established chemical structures allows receiving adequate,

According to the multifunctional potential of artificial GC used, resulting events in biotope can be the following ones: antipathogenic actions (the use of GC-imitating pathogenic cell surface structures), prebiotic and symbiotic (synbiotic) actions (the use of GC-imitating prebiotic structures), increasing own human organism protective systems (the use of GC-interacting and GC-regulating macrophages and macrophage-like lymphocytes through their systemic receptor lectins), antitumor actions (potential using antigenic GC together with LSSM as antagonists and synergists in intercellular antitumor and anti-infectious communications [37]), and cytokine lectin/lectin-like cascades initiated/regulated with LSSM potentially

**3.6 Prospects of LSSM for prophylaxis and therapy of infectious diseases**

Innate immunity plays an important role especially when antibody immunity is under development, suppression, or alteration. So the search of new natural antiinfectious agents is of actuality. In contrast to probiotic cells, LSSM (as systems of LL or LB) are not sensitive to the presence of antibiotics and chemotherapeutics as well as do not need special conditions for survival. LSSM function as metabolomebiotics according to the principle "the network in the network"; participation of major lectins in supporting physiologically significant glycodecor of biotope mucosal probiotic compartments, and minor lectins—in signal communications; support of such compartments (compensation of the absence of probiotic cells in MB upon therapy with antibiotics, delivery of fucosylated and galactosylated prebiotics of mutual supporting influence between populations of bifidobacteria and lactobacilli,

influencing redistribution of cytokine network in organism.

(pI 3–4) serial phyto-oxidoreductases/phyto[glycosyl]oxidases"].

#### *Metabolite Multiprobiotic Formulas for Microbial Health DOI: http://dx.doi.org/10.5772/intechopen.86449*

*Oral Health by Using Probiotic Products*

body of pathogen) for the mostly effective visible actions of LS, obtaining uniform pure landscapes of LSSM action on massif of *Candida albicans* (the absence or minimization of LSSM-resistant residual fungal colonies in the interacting intestine system "LS of human intestinal bifidobacteria and lactobacilli—human intestinal *C. albicans*"). Antimicrobial activities of LSSM and phytolectins (phytohemagglutinin from the kidney bean) could be realized not only directly but also through the influence (together with synthetic mannans and mucins) in respect to macrophage migration as well as through inducing production of cytokines by stimulated blood lymphocytes (on the example of tumor necrosis factor-α). Results indicate prospects of anaerobic LSSM as assistant ingredients of the possible drug forms.

During the last time, synbiotics and symbiotics (as synergistic sum of probiotics and prebiotics) are of increased investigator interest due to their antimicrobial and other useful reactions [1, 2]. In this respect LSSM represent new class of antipathogenic proteins (possessing extended potential of application) which recognize different GC. LSSM represent multifunctional potential of relatively highly molecular mass polymeric metabolites of cultures of the human microbiota (microbiocenoses) and consortia (also multistrain probiotics) of human indigenous microorganisms. LSSM cofunction together with artificial and biologically active natural GC [32]. According to own results, we proposed suitable laboratory minisystem for screening prebiotic and therapeutical GP using LSSM and sterile heparinized insulin syringes of 1 ml volume. The following results were obtained: (1) LSSM-containing fraction stimulated production of both the whole and adhesive mass of bifidobacteria, (2) LiCl (15 mM and higher) increased dose depending on the number of adhesive colonies, (3) bifidobacterial LSSM (within pI 4–4.5) were characterized with strong affinity to anionic synthetic GC (possessing exposed residues of sulfated galactosides or, in a less extent of affinity, exposed residues of mannose-6-phosphates), and

participated in functioning bioreactor imitating synbiotope (multiplication

Proposed synbiotic system is perspective for screening prebiotic GC (as it is known for prebiotic derivatives of chitin, chitosan, fucoidan, and glycopeptides [33, 34]). **Table 1** includes potential prebiotic sources such as chitin and α-L-fucan which react to LSSM (LSSM may serve carriers of metabiotic GC; the list of GC can

Progress in membrane and solid-phased technologies using LS-GC interactions includes the potential of their application in microassays, biochips (membrane

The use of affine pore hydrophobic (uncharged) membranes predictably covered with mosaics of multifunctional sets of LSSM (additional significant purification of LSSM on membranes is reached) allows prolonged storing LSSM without decreasing samples in activities. The following prospects of LSSM-GC combinations may be of practical interest: (a) antifungal covers of prolonged action in combination with antibiotics and physical factors of stress [radiation (ultraviolet and ultrasonic), light (biorhythm "day-night"), temperature, pH, oxygen, season changes (also biorhythmic), others] and (b) chemiluminescent systems cofunctioning in regime of real time for medical and industrial biotechnology and

and LSSM as potential car-

**3.4 Synbiotic minibioreactor using LSSM for screening GC**

(4) sulfated glycosaminoglycans together with cations Li+

**3.5 Membrane technological prospects of LSSM**

of bifidobacterial colonies and their survival were observed).

bound glycoarrays or lectin arrays), and biosensors [35, 36].

**82**

riers of Li+

be unlimitedly extended).

bionanotechnologies [our results include the following coupled systems: "low acidic LSSM, low acidic oxidoreductases of lactobacilli"; "alkaline bifidobacterial LS, alkaline bifidobacterial exopolymeric compounds"; "neutral lactobacillar/bifidobacterial LS, neutral lactobacillar/bifidobacterial biosurfactants"; "LS, strongly acidic (pI 3–4) serial phyto-oxidoreductases/phyto[glycosyl]oxidases"].

Membrane technologies using separated proteins, oligopeptides, and their complexes (especially) together with intrinsic or exogenic (SYPRO dye) fluorescence registered in live bioimagination are especially sensitive and perspective (protein band discreteness using fluorescence technique was better compared to the chemiluminescence technique). The latter allows identification stabile boundaries of the whole protein massifs for further establishment of LSSM and other biologically and physiologically active components among protein mosaics. Bioluminescence (fluorescent technique in combination with chemiluminescence technique) in optimal (depending on the object and the goal of study) conditions allows express-ranging cultural fluid groups of proteins and LS according to molecular mass (for additional standardization and typing of strains), evaluation of interstrain synergism and contribution of protease and oxidoreductase systems of mono- and multistrain probiotics (symbiotics) and other type consortia, and identification of mosaics of complexes containing fluorochromes in extended interval of pI/pH (complexes and cell wall fragments as carriers of visible energy which is ready for energetic exchange with surrounding infrastructures as well as for monitoring directed supramolecular assembling and their reorganization).

Aforementioned data presented develop other possible important prognostic approaches and proposals. The choice of wished prognostic (LSSM-selected-type and GC-type interactions)-directed events in biotope can be determined and regulated by involving GC types needed (panels of natural and artificial GC used in biotope; extending the list of GC indicated in **Table 1** is possible). The use of artificial polymeric GC with established chemical structures allows receiving adequate, understandable, and reliable results.

According to the multifunctional potential of artificial GC used, resulting events in biotope can be the following ones: antipathogenic actions (the use of GC-imitating pathogenic cell surface structures), prebiotic and symbiotic (synbiotic) actions (the use of GC-imitating prebiotic structures), increasing own human organism protective systems (the use of GC-interacting and GC-regulating macrophages and macrophage-like lymphocytes through their systemic receptor lectins), antitumor actions (potential using antigenic GC together with LSSM as antagonists and synergists in intercellular antitumor and anti-infectious communications [37]), and cytokine lectin/lectin-like cascades initiated/regulated with LSSM potentially influencing redistribution of cytokine network in organism.

#### **3.6 Prospects of LSSM for prophylaxis and therapy of infectious diseases**

Innate immunity plays an important role especially when antibody immunity is under development, suppression, or alteration. So the search of new natural antiinfectious agents is of actuality. In contrast to probiotic cells, LSSM (as systems of LL or LB) are not sensitive to the presence of antibiotics and chemotherapeutics as well as do not need special conditions for survival. LSSM function as metabolomebiotics according to the principle "the network in the network"; participation of major lectins in supporting physiologically significant glycodecor of biotope mucosal probiotic compartments, and minor lectins—in signal communications; support of such compartments (compensation of the absence of probiotic cells in MB upon therapy with antibiotics, delivery of fucosylated and galactosylated prebiotics of mutual supporting influence between populations of bifidobacteria and lactobacilli, cofunctioning to anti-infectious agents (antibiotics, metabiotics, GC) and cells of organism protective systems (macrophages and leukocytes).

We characterize LSSM (acidic, low acidic, cationic: aLL, laLL, cLL, aLB, and cLB) of domestic probiotics which recognize polymeric artificial GC mucin-like analogs of antigens and EPC (fungal and bacterial mannans, bifidobacterial α-L-Fuc-containing, sulfated and phosphorylated).

*Antimicrobial directions of LSSM action* were established [26, 31, 38].
