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The role of mucoid polysaccharide (Hyaluronic acid) in the virulence of group A hemolytic Streptococci
Abstract
1. A quantitative turbidimetric method for the estimation of hyaluronidase activity, based on the ability of the enzyme to decrease the capacity of the polysaccharide to precipitate acidified protein has been developed. Two units of hyaluronidase, by this method, are equivalent to one viscosity-reducing unit. 2. Hyaluronidase added to a phagocytic system containing defibrinated human blood, immune or non-immune, greatly increases the rate of phagocytosis of group A streptococci. Phagocytosis of Type I pneumococci is not affected by hyaluronidase under the same conditions. 3. The bactericidal activity of non-immune blood against group A streptococci is increased by hyaluronidase; the activity of immune blood is, however, somewhat inhibited by the enzyme. Killing of pneumococci is not affected by the presence of the enzyme. 4. Mice can be protected against group A streptococcal infection by frequent treatment with 200 turbidity-reducing units of hyaluronidase; the protective action of the enzyme is removed by heating at 60 degrees C. for 1 hour. Mice infected with Type I pneumococcus and treated with hyaluronidase die somewhat sooner than the untreated controls.
... Hyaluronidase inhibition activity was determined turbidimetrically by the method of Kass et al [22] by using 0.01 mg/ml enzyme mixed with 250 µg/ml from the extracts with inhibition time 45 min. and the percentage of inhibition %I was calculated according to this equation [23]: The biological activity against various bacterial species was determined. ...
... Such compounds had been reported to have an active effect on the bacterial cells membrane, which may caused destroy these microorganisms [7,18]. The alkaloids interact with the DNA, the tannins inhibit the carrier enzymes and proteins present in the cells membrane, while the phenolic compounds form complex with dissolved protein out of the cells or with cells membrane which made to destroy the bacteria [18,22]. ...
Natural plant Dandelion Taraxacum officinale have been used as a phytomedicine, in this study, the chemical components of the Dandelion Taraxacum officinale leaves in watery and alcoholic extracts were identified. The results showed that watery extract was alkaline (presence of alkaloids) while the alcoholic extract was acidic in these contained: glycosides, alkaloids, phenolic compounds, tannins, flavonoids and proteins, while the saponins and resins were not found. The result also showed that high concentrations of the following trace elements were found in the leaves (K ,Ca , Na ,Fe) with 185.1, 22, 19.5, 11.2 ppm, respectively and low concentrations of (Zn, Cd, Cu) with 6.3, 1.3, 0.2 ppm, respectively. The effect of these extracts on the different microorganisms were studied. It has been found that the concentration 0.5 mg/ml was effective on the inhibition of the growth of the intended bacteria (for both extracts) especially Gram positive, Staphylococcus aureus and the alcoholic extracts with concentration 0.5 mg/ml was more effective on the Gram negative, E.coli, than the watery extract, while less than 0.1 mg/ml failed to inhibit any microorganisms. The High Performance Liquid Chromatography (HPLC) was used to identify some flavonoids as compared to standard one; the analysis showed that both kaempferol and morin were absent.
... Studies have been carried out by making modifications on a method based on the clouding of hyaluronic acid and albumin solution. [44,45] In the 96-well microplate, 20 ml of each plant extract (or nanoflower of the extract) was added and 10 ml of hyaluronic acid solution (concentration of 0.4 mg/ mL) was added and incubated at 37 C for 4 À 5 minutes to ensure the temperature balance. Then 10 ml of hyaluronidase enzyme solution (at concentration of 0.05 mg/mL) is added and incubated for 10 minutes. ...
This study aimed to evaluate the hyaluronidase and gelatinase inhibitory activities and toxicity potentials of Persea americana Mill. extract and its synthesized hybrid nanoflowers. In the first part of the study, characterization of the nanoflower structures was carried out and then, enzyme inhibitory activities of the methanol extract and hybrid nanoflowers were investigated. It was determined that the enzyme inhibitory activity of both nanoflowers was higher than the methanol extract. Zinc nanoflower was more effective in hyaluronidase enzyme inhibition, while copper nanoflower showed higher inhibitory activity on gelatinase enzyme. In the second part of the study, toxicological profiles of these compounds were investigated. Toxicological evaluations demonstrated that zinc nanoflower may be a safer therapeutic alternative than the copper nanoflower, especially at high concentrations. All these data appear to contribute to the development of effective new generation preparations including nanoflowers for skin problems using Persea ameri-cana leaves extract and its nanostructures. ARTICLE HISTORY
... The resistance of Group A streptococci to phagocytosis by blood leucocytes has been shown to be related to the presence of at least two surface components, M protein and hyaluronic acid capsules (3)(4)(5)(6)(7). In the absence of these components, Group A organisms are opsonized by the bloods of all mammalian species studied. ...
In the article by Weissmann and Thomas on Studies on lysosomes. I, in Text-figs. 1, 2, and 3, and in Tables I, II, III, and IV, for /micrograms protein/hour read everywhere /100 micrograms protein/hour .
... The strains of bacteria that produce HA, incorporate the newly synthesized HA to the bacterial mucoid capsule, which confers to the bacteria resistance to opsonization, immune camouflage and protection against the host immune system [35,36]. Kass and Seastone have demonstrated that when group A and C streptococci have their mucoid capsule degraded by hyaluronidases, they decreased their infectiousness in mice, as their susceptibility to phagocytosis by leukocytes increase [37]. Other studies have confirmed the association between HA capsule production and virulence by deletion or mutation of the bacterial Fig. 1. ...
The relationships between hyaluronic acid (HA) and pathological microorganisms incite new understandings on microbial infection, tissue penetration, disease progression and lastly, potential treatments. These understandings are important for the advancement of next generation antimicrobial therapeutical strategies for the control of healthcare-associated infections. Herein, this review will focus on the interplay between HA, bacteria, fungi, and viruses. This review will also comprehensively detail and discuss the antimicrobial activity displayed by various HA molecular weights for a variety of biomedical and pharmaceutical applications, including microbiology, pharmaceutics, and tissue engineering.
... Hyaluronidase inhibition activity was determined turbidimetrically by the method of Kass et al. [24] by using 0.01 mg/ml enzyme mixed with 250 µg/ml from the extracts with inhibition time 45 min. and the percentage of inhibition was calculated according to this equation [25]: ...
Mirabilis jalapa , المركبات من مجموعة على يحتويان والكحولي المائي المستخلصين محلولي أن الدراسة أظهرت إذ والبروتينات والقلويدات والراتنجات والفينولية والعفصية الكاليكوسيدية بينما الصابونيات على تحتوي ال. والفالفونويدات المعد للعناصر الدقيق التحليل اثبت من تراكيز على احتواءها النبات ألوراق نية K و Na و Fe وهي 161.2 ,19 18.7 µg/ml من وكميات , التوالي على Zn و Ca وهي 12, 14.2µg/ml على وكميات Cd , Cu و Pb وهي 0.8 ,0.3,0.1 µg/ml. و التوالي على على احتوائها وعدم , Cr. للتركيز أن لوحظ أذ البكتريا من مختلفة أنواع على والكحولية المائية المستخلصات تأثير درس كما 0.5 بكتريا نمو تثبيط تجاه ّاال فع تأثيرا /مل ملغم Proteus mirabilis و E.coli و Staphylocoous aureus. الكحولي المستخلص تجاه خاصة ABSTRACT The chemical components of the Mirabilis jalapa L. leaves in the watery and alcoholic extracts were identified .The results showed that the extract contain : glycosides ,tannins ,phenolic compounds , resins ,alkaloids and proteins ,while the saponins and flavonoids were not found. The results also showed that there were concentrations of the following trace elements in the leaves K , Na , Fe with 161.2 ,19 ,18.7 µg/ml , respectively and concentrations of Zn ,Ca with 14.2 , 12 µg/ml respectively , concentrations Cd ,Cu, Pb with 0.8 ,0.3,0.1 µg/ml respectively ,and Cr were not founds. The effects of these extracts on growth of different bacteria were studied, has been found that 0.5 mg/ml concentration was effective inhibitor of growth of Staphylocoous aureus , E.coli and Proteus mirabilis .
... A turbidimetric method was used to determine the hyaluronidase enzyme activity as describe previously (Tolksdorf et al., 1949 Kass andSeastone, 1944). Briefly, venom samples with the following venom concentrations; 1, 2.5, 5, 7.5, 10, 15, 20, and 25 mg/ ml were diluted in 50 ml of sodium acetate buffer (0.2 M sodium acetate, 0.15 M NaCl, pH 6.0). ...
... The GAS capsule has been studied extensively since its initial description by Bordet in 1907 (29). Very early studies demonstrated that the GAS capsule is composed of hyaluronic acid, is nonimmunogenic, and is a major virulence factor contributing to resistance to phagocytosis by human polymorphonuclear leukocytes (PMNs) (29)(30)(31). Subsequently, the hasA and hasB genes were shown to encode enzymes required for capsule biosynthesis (32). The frameshift mutation in strain MGAS12503 results in a premature stop codon that truncates the HasA protein by 104 amino acids ( Fig. 2A). ...
Humans commonly carry pathogenic bacteria asymptomatically, but despite decades of study, the underlying molecular contributors
remain poorly understood. Here, we show that a group A streptococcus carriage strain contains a frameshift mutation in the
hasA gene resulting in loss of hyaluronic acid capsule biosynthesis. This mutation was repaired by allelic replacement, resulting
in restoration of capsule production in the isogenic derivative strain. The “repaired” isogenic strain was significantly more
virulent than the carriage strain in a mouse model of necrotizing fasciitis and had enhanced growth ex vivo in human blood. Importantly, the repaired isogenic strain colonized the mouse oropharynx with significantly greater bacterial
burden and had significantly reduced ability to internalize into cultured epithelial cells than the acapsular carriage strain.
We conducted full-genome sequencing of 81 strains cultured serially from 19 epidemiologically unrelated human subjects and
discovered the common theme that mutations negatively affecting capsule biosynthesis arise in vivo in the has operon. The significantly decreased capsule production is a key factor contributing to the molecular détente between pathogen
and host. Our discoveries suggest a general model for bacterial pathogens in which mutations that downregulate or ablate virulence
factor production contribute to carriage.
... Anti-hyaluronidase activity was determined by the turbidimetric method suggested by Kass and Seastone [49] in which hyaluronic acid reacts with albumin acid solution. This turbidity is a function of hyaluronic acid concentration and can hence be related to enzyme activity. ...
Safranal, a monoterpene aldehyde, is present as one of the main volatile constituents of Crocus sativus Linn. (saffron flowers). This volatile constituent not only contributes to the aroma of saffron but has been reported to possess antidiabetic, antiulcer, antiasthamatic, anticonvulsant, antidepressant, cardioprotective, anticancer and UV protective properties. Most of these therapeutic actions are contributed by its potential to quench reactive oxygen species (ROS). Antioxidant properties of phytoconstituents are now being explored for developing photoprotective skin formulations. These bioactives have the potential to protect the epidermal and dermal layers of the skin which mainly comprises of elastin and collagen. When UV rays penetrate the dermal layers, there is an increased production of elastase, collagenase and hyaluronidase leading to degradation of collagen, elastin and hyaluronic acid respectively. These dermal components are responsible to provide strength, elasticity and moisture to the skin. Due to frequent exposure to sunlight, these conditions tend to augment leading to wrinkle formation and sagging of skin.
Although antioxidant properties of safranal have been established on various cell lines but till date no studies have been reported regarding the dermal enzyme inhibition activities. In the current research work, a comprehensive in vitro evaluation of antioxidant, anti-elastase, anti-collagenase, anti-hyaluronidase activities of safranal along with determination of sun protection factor (SPF) was carried out. The in vitro antioxidant activity was carried out by diphenylpicrylhydrazyl (DPPH) method and its IC50 value was found to be 22.7 μg/ml. The enzyme inhibition IC50 values of safranal for anti elastase activity were found to be 43.6 μg/ml, 70 μg/ml for antihyaluronidase activity and 9.4 μg/ml for anticollagenase activity. Photoprotective activity of safranal was determined by UV absorbance method and SPF calculated by Mansur equation which was found to be 6.6.
The significant inhibitory activity of safranal on matrix metalloproteinases (MMPs) responsible for aging and a higher SPF established that this bioorganic molecule is a strong photoprotective agent. Its established free radical scavenging capability along with above characteristics make it a valuable component to be incorporated into herbal antiaging formulations.
... At specified intervals samples in a volume of 0.5 mL were taken from the reaction mixture and added into suitable cuvettes containing 2.5 mL of Acidic Bovine Albumin Solution (24 mM sodium acetate, 79 mM acetic acid with 0.1% (w/v) bovine serum albumin, pH 3.75) and mixed immediately. The method [37,38] is based on the ability of HA (as polymer) to form a complex with serum albumin in an acidic environment. Appearing turbidity is a function of the HA polymer concentration and thus can be associated with enzyme activity. ...
Polymeric drug delivery system, allowing the controlled long-term drug release, is a good alternative to the standard way (oral or intravenous) of pharmaceuticals' administration. The most widespread drug release strategy is based on the drug molecules diffusion: first inside the carrier, then through the external layers of carrier and then, outside the carrier, to the cells surrounding. The proposed type of the local therapy is a particularly proper approach in treatment of cancer diseases, at which the drugs are highly toxic. Unfortunately, antitumor drugs diffusion is quite often limited by hyaluronic acid layers abundantly found in a cancer tissue. The undertaken approach assumed relaxation of hyaluronan layer by using a hydrolytic enzyme - hyaluronidase. The considered process corresponds to the heterogenic catalysis with substrate occurred as a layer and a catalyst in a native form. It was shown that enzyme concentration around 4 mg/L kept for 10 minutes in body fluid surrounding HA layers is enough to allow the drug to diffuse. Hydrogel carriers promoting fast release of hyaluronidase were characterized. An expected rate value (the mass transport coefficient app. 4.0.10-7 m/s) was obtained for blends 0.5% κ-carrageenan, 4% polyvinyl alcohol and 0.5% sodium alginate, 6% polyvinyl alcohol.
... Hirst found that decapsulating S. pyogenes by treatment with leech extract containing hyaluronidase had no effect on virulence of the bacteria in mice (77). However, Kass and Seastone found reduced virulence of decapsulated S. pyogenes in mice, perhaps because the latter authors administered hyaluronidase to the infected mice at frequent intervals in order to prevent regeneration of the capsule after challenge (78). Subsequently, more direct proof that the capsule contributes to virulence has come from several studies using acapsular mutant strains of S. pyogenes derived by transposon mutagenesis or by targeted inactivation of gene(s) required for hyaluronic acid synthesis (Table). ...
Capsular Polysaccharide of Group A Streptococcus, Page 1 of 2
Abstract
Most clinical isolates of Streptococcus pyogenes elaborate a capsular polysaccharide, which is composed of hyaluronic acid, a high-molecular-mass polymer of alternating residues of N-acetyl glucosamine and glucuronic acid. Certain strains, particularly those of the M18 serotype, produce abundant amounts of capsule, resulting in formation of large, wet-appearing, translucent or “mucoid” colonies on solid media, whereas strains of M-types 4 and 22 produce none. Studies of acapsular mutant strains have provided evidence that the capsule enhances virulence in animal models of infection, an effect attributable, at least in part, to resistance to complement-mediated opsonophagocytic killing by leukocytes. The presence of the hyaluronic acid capsule may mask adhesins on the bacterial cell wall. However, the capsule itself can mediate bacterial attachment to host cells by binding to the hyaluronic-acid binding protein, CD44. Furthermore, binding of the S. pyogenes capsule to CD44 on host epithelial cells can trigger signaling events that disrupt cell-cell junctions and facilitate bacterial invasion into deep tissues. This article summarizes the biochemistry, genetics, regulation, and role in pathogenesis of this important virulence determinant.
... Studies of the GAS capsule spanning the past century seemingly solidified its role as a key GAS virulence factor [7]. The GAS capsule consists of hyaluronic acid and is a major contributor to resistance to phagocytosis by human neutrophils [8][9][10]. However, results of studies have suggested that the capsule might be dispensable under certain conditions (eg, asymptomatic carriage) [11]. ...
Background:
Bacterial infections caused by group A Streptococcus (GAS) are common in childhood. Few study reports have provided data on pediatric-specific trends in the epidemiology and bacterial strain characteristics of GAS infections.
Methods:
We prospectively collected GAS isolates from the clinical microbiology laboratory at Texas Children's Hospital between July 1, 2013, and June 30, 2017. Patient characteristics and GAS disease categories were determined through chart review. GAS isolates were obtained from patients in either the inpatient or outpatient setting, and cases were defined as pharyngeal disease, skin and soft-tissue infection (SSTI), or invasive disease on the basis of predefined criteria. All isolates were emm typed to determine trends over time.
Results:
We identified 930 cases over the 4-year period, including 432 (46.4%) pharyngeal, 235 (25.3%) SSTI, and 263 (28.3%) invasive disease types. The most frequently encountered emm types were emm1 (21.4%), emm12 (15.7%), emm89 (14.6%), emm4 (9.2%), and emm3 (8.2%). We observed significant changes over the 4-year period in the relative frequency of infections caused by emm1 (-17.7%; P = .046), emm4 (8.7%; P = .023), or emm6 (-7.9%; P = .024). Using bioinformatic analyses and targeted gene sequencing, we also discovered that all GAS emm28 and emm87 types harbored mutations that rendered them incapable of producing capsule. The relative frequency of GAS disease cases caused by capsule-negative GAS emm types (emm4, emm22, emm28, emm87, and emm89) increased over the 4-year period (32.2%-44.4%), although the difference was statistically significant for only nonpharyngeal disease types (27.1%-43.9%; P = .038).
Conclusions:
Our data suggest an evolving epidemiology of GAS in the Houston pediatric population characterized by an increase in the frequency of capsule-negative emm types.
... A major virulence factor in the pathogenesis of S. pyogenes is the capsule that is composed of hyaluronic acid, the same component in human connective tissues. The capsule contributes to the pathogenesis of S. pyogenes as an antiphagocytic factor [1][2][3] and adhesin to keratinocytes [3][4][5][6]. ...
A hyaluronic acid capsule is a major virulence factor in the pathogenesis of Streptococcus pyogenes. It acts as an anti-phagocytic agent and adhesin to keratinocytes. The expression of the capsule is primarily regulated at the transcriptional level by the two-component regulatory system CovRS, in which CovR acts as a transcriptional repressor. The covRS genes are frequently mutated in many invasive strains, and a subset of the invasive CovRS mutants does not produce a detectable level of the capsule at 37 °C, but produces a significant amount of the capsule at sub-body temperatures. Here, we report that a prophage has a crucial role in this capsule thermoregulation. Passaging CovR-null strains showing capsule thermoregulation using a lab medium produced spontaneous mutants producing a significant amount of the capsule regardless of incubation temperature and this phenotypic change was caused by curing of a particular prophage. The lab strain HSC5 contains three prophages on the chromosome, and only ϕHSC5.3 was cured in all spontaneous mutants. This result indicates that the prophage ϕHSC5.3 plays a crucial role in capsule thermoregulation, most likely by repressing capsule production at 37 °C.
... The mechanism by which the M antigen permits streptococcal cells to resist phagocytosis has not been explained. Moreover, in addition to the M protein, the hyaluronic acid capsule has been shown to contribute a measurable degree of nonspecific resistance to phagocytosis by group A streptococci (37,38,84). Cinephotomicrography studies by the late Armine Wilson (178) demonstrated what appeared to be surface phenomena enabling virulent group A streptococcal chains to slither off the surfaces of leucocytes vainly attempting to engulf them. ...
... One of the first pieces of evidence showing that some bacterial strains produce HA was shown by the study of Kendall, Heidelberger and Dawson, showing the isolation of a polysaccharide from the culture media of Streptococcus haemolyticus group A, which was successively identified as HA [41]. From a structural viewpoint, HA can be found in the capsule of other Streptococcus species belonging to group A and C [42][43][44] and in the capsule of Gram-negative Pasteurella multocida [45], allowing them to escape or overcome the host cellular and humoral immune responses since the capsule does not trigger the immune response of the host, prevents macrophage phagocytosis and enhances the virulence of these bacteria. HA concurring to the formation of S. pyogenes capsule is characterized by its HMW size and can be produced by incubation of cell-free membrane extracts of S. pyogenes with the UDP-sugars in the presence of divalent cations [46,47]. ...
The commensal microbiota plays a fundamental role in maintaining host gut homeostasis by controlling several metabolic, neuronal and immune functions. Conversely, changes in the gut microenvironment may alter the saprophytic microbial community and function, hampering the positive relationship with the host. In this bidirectional interplay between the gut microbiota and the host, hyaluronan (HA), an unbranched glycosaminoglycan component of the extracellular matrix, has a multifaceted role. HA is fundamental for bacterial metabolism and influences bacterial adhesiveness to the mucosal layer and diffusion across the epithelial barrier. In the host, HA may be produced and distributed in different cellular components within the gut microenvironment, playing a role in the modulation of immune and neuronal responses. This review covers the more recent studies highlighting the relevance of HA as a putative modulator of the communication between luminal bacteria and the host gut neuro-immune axis both in health and disease conditions, such as inflammatory bowel disease and ischemia/reperfusion injury.
... Avery and Dubos (15) authored one of the earliest reports of the therapeutic use of enzymes to treat infections and the first use of capsule-degrading enzymes in the 1930s. Relatively crude preparations of capsule-degrading enzymes were used to remove the capsular polysaccharides from Streptococcus pneumoniae and were shown to be effective in treating experimentally infected mice, rabbits, and monkeys (15)(16)(17)(18)(19). Similar therapeutic approaches to remove capsules enzymatically to treat infections with bacterial and fungal pathogens have been reported for groups C and A Streptococcus, Cryptococcus neoformans, Escherichia coli, and Klebsiella pneumoniae (20)(21)(22)(23). These enzymes do not kill the bacteria but render them susceptible to the host innate immune responses by removing the antiphagocytic capsule. ...
Anthrax is considered one of the most dangerous bioweapon agents, and concern about multidrug-resistant strains has led to the development of alternative therapeutic approaches that target the antiphagocytic capsule, an essential virulence determinant of Bacillus anthracis, the causative agent. Capsule depolymerase is a γ-glutamyltransferase that anchors the capsule to the cell wall of B. anthracis. Encapsulated strains of B. anthracis can be treated with recombinant capsule depolymerase to enzymatically remove the capsule and promote phagocytosis and killing by human neutrophils. Here, we show that pegylation improved the pharmacokinetic and therapeutic properties of a previously described variant of capsule depolymerase, CapD-CP, when delivered 24 hours after exposure every 8 hours for 2 days for the treatment of mice infected with B. anthracis. Mice infected with 382 LD50 of B. anthracis spores from a nontoxigenic encapsulated strain were completely protected (10 of 10) after treatment with the pegylated PEG-CapD-CPS334C, whereas 10% of control mice (1 of 10) survived with control treatment using bovine serum albumin (P < 0.0001, log-rank analysis). Treatment of mice infected with five LD50 of a fully virulent toxigenic, encapsulated B. anthracis strain with PEG-CapD-CPS334C protected 80% (8 of 10) of the animals, whereas 20% of controls (2 of 10) survived (P = 0.0125, log-rank analysis). This strategy renders B. anthracis susceptible to innate immune responses and does not rely on antibiotics. These findings suggest that enzyme-catalyzed removal of the capsule may be a potential therapeutic strategy for the treatment of multidrug- or vaccine-resistant anthrax and other bacterial infections.
... Based on the studies by Tolksdorf et al., Kass and Seastone, and Muckenschnabel et al., HA degradation was measured via the effect of the addition of the HYL enzyme on turbidity (Tolksdorf et al., 1949; Kass and Seastone, 1944; Muckenschnabel et al., 1998). Turbidity is a function of HA concentration and can be related to enzyme activity. ...
... These bioactive compounds show antibacterial effects through different mechanisms. For example, saponins have been reported to play a part in managing inflammation (Adebayo and Issah, 2012); tannins work by reacting with proteins has been proved to provide a tanning effect necessary for treating inflammation and inhibiting carrier enzymes and proteins in cell membranes (Kass and Seastone, 1944;Parekh and Chanda, 2007); alkaloids are able to interact with the DNA, and the phenolic compounds formed complex with dissolved protein out of the cells or with cell membranes to kill bacteria (Nikaido, 1994;Hu and Kitts, 2005;Jassim et al., 2012). In other related reports, alpha-tocopherols were reported to destroy the efflux pumps of S. aureus to reduce resistance (Tintino et al., 2016), and organic acids were able to make the bacterial growth environment more acidic, and inhibit the formation of capsular polysaccharide of bacteria to exert antibacterial effects (Chen et al., 2011;Lin et al., 2013;Mitani et al., 2018). ...
Due to the dramatic increase in the use of antibiotics and growing health threat of bacterial resistance to many commonly used antibiotics, many studies have been directed at developing new and effective antibacterial compounds, among which many new, natural, and effective antibacterial compounds discovered from medicinal plants have drawn great interest and raised new hope for treating the challenges of antibiotic resistance. This review aimed to summarize the most important and widely used medicinal plants that were reported to have antibacterial activities. A general literature search from 2010 to 2020 was conducted using different databases, including Science Direct, Web of Science, and PubMed. According to the literature, three medicinal plants with outstanding antibacterial activities, Taraxacum officinale, Coptis Rhizome, and Scutellaria baicalensis, were screened and reviewed by prioritization. The extraction methods, antibacterial activities of different parts of plants or the plant-derived compounds, spectra of antibacterial activities, and toxicity were described, respectively. However, the antibacterial activities of the extracts or pure compounds as reported in the reviewed literature were mostly based on in vitro assays, and moreover, the deeper antibacterial mechanisms have not been elucidated clearly. Therefore, further studies are required in the fields of purification and identification of the antibacterial compounds, its mechanisms of action, and synergistic effects in combination with other antibacterial drugs, which may be helpful in the development of new antibacterial drugs.
... Pathogens and microbes of our normal microbiome synthesize capsules 32,33 or cell walls rich in polysaccharides and mannosylated proteins, 34 The bars represent means ± SEMs. See also Figure S4 demonstrating that force overcomes electrical barriers to particle engagement. ...
Macrophages continuously survey their environment in search of pathogens or apoptotic corpses or debris. Targets intended for clearance expose ligands that initiate their phagocytosis (“eat me” signals), while others avoid phagocytosis by displaying inhibitory ligands (“don’t eat me” signals). We report that such ligands can be obscured by the glycosaminoglycans and glycoproteins that coat pathogenic as well as malignant phagocytic targets. In addition, a reciprocal barrier of self-synthesized or acquired glycocalyx components on the macrophage surface shrouds phagocytic receptors, curtailing their ability to engage particles. The coating layers of macrophages and their targets hinder phagocytosis by both steric and electrostatic means. Their removal by enzymatic means is shown to markedly enhance phagocytic efficiency. In particular, we show that the removal of mucins, which are overexpressed in cancer cells, facilitates their clearance. These results shed light on the physical barriers that modulate phagocytosis, which have been heretofore underappreciated.
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Hyaluronidase is an enzyme has a great potential in diverse medical fields. A group of bacterial isolates was screened for the production of hyaluronidas. The most potent isolate was identified depending on of its morphological, biochemical and molecular characteristics. Thereafter, the new powerful enzyme producer was identified as Staphylococcous aureus. Optimization strategy was conducted to enhance the enzyme production by S. aureus in a medium containing a waste material (milk whey). The first step of the optimization process included choosing of the most significant factors after screening with Plackett-Burman model. Eleven factors, medium composition and some cultural conditions, were screened. Hence, the most significant factors were subjected to further optimization step using Box-Behnken model. Box-Behnken model had the ability to suggest the optimum concentrations of the significant parameters and to predict the maximal enzyme activity. The predicted enzyme activity was 500 U/ml. The predicted value was verified experimentally, and the maximum enzyme activity reached 492U/ml. Overall, the final enzyme outcome 492 U/ml is considered absolutely high if it compared with all previous findings with free cells. In addition to using of competitive cheaper media, attempts were made to increase the effectiveness of the enzyme through partial purification.
1.1. Aphonopelma chalcodes and Dugesiella hentzi tarantulas were successively milked at different intervals and the venoms were analyzed quantitatively.2.2. Multiple successive milkings at defined intervals resulted in relatively constant venom yields.3.3. The amount of venom regenerated was relative to the time interval between successive milkings.4.4. Regeneration of the analyzed venom constituents was not completely uniform: proteins, free amino acids and toxicity reached more or less the same levels as fresh venom, but hyaluronidase was resynthe-sized at a considerably slower rate.5.5. Seasonal variations in the speed of venom regeneration were observed.6.6. The similarity of tarantula venom regeneration to those of snake venoms is discussed.
Bacteria, which are among the most numerous members of the microbiologic population, are at the same time microscopic plant cells and individual organisms. Because of the practical importance of bacteria in medicine, agriculture, and industry, bacteriology is cultivated mainly as an applied science. This chapter discusses the aspects of bacteria as cells and as organisms. The observations of genetic behavior in bacteria include the phenomena of the continuity of genetic determiners, their recombination and segregation, which are currently expressed in the idioms of classical genetics. Analysis of nuclear phenomena in bacteria by the most revealing techniques presently available is exhibiting the sequences of classical mitosis. Bacterial cells conform the familiar pattern of cellular organization, in which they possess external cell wall, cytoplasmic membrane, and protoplast containing well-differentiated nucleus and organelles for organized energy exchange or mitochondria. The fundamental biochemical similarities between the cells of bacteria and of higher plants and animals are also discussed.
The vascular permeability-increasing action of rabbit PMNL lysosomes has been studied in skin and cremaster muscle of the rat. Both an extract of frozen-thawed granules and a cathepsin-free cationic protein fraction of the granules (which had previously been demonstrated to cause leukocyte adhesion and emigration in vivo) induce increased vascular permeability in skin and muscle which resembles that produced by histamine or histamine-liberators with respect to the timing of the response and the predominant type of microvessel affected. Extracts of frozen-thawed lysosomes and the inflammatory lysosomal cationic protein both cause disruption of rat mesenteric mast cells in vitro, whereas a granule-free cytoplasmic fraction of PMN leukocytes and a non-inflammatory cationic protein fraction of the granules do not do so under identical test conditions. The mastocytolytic action of lysosomal materials in vitro is not inhibited in the presence of 10 kallikrein-inhibiting units of trasylol per ml. The mast cell rupturing fraction of PMNL granules (cationic protein) possesses no detectable peroxidase activity or acid-mucopolysaccharase activity. When compared with compound 48/80 on the basis of estimated molecular weight, the lysosomal cationic protein appears to be at least as active as the latter compound with respect to in vitro mastocytolytic potency. Chronic pretreatment of rats with an agent known to reduce tissue mast cell numbers causes marked suppression of the vascular permeability change normally induced in skin and muscle by lysosomal extracts and cationic protein. Similar results are obtained if lysosomal materials are tested in rats pretreated with an antihistaminic. These observations are discussed with respect to the mode of action of PMNL lysosomes in the early and late phases of local tissue-injury reactions.
Antigene und Antikörper sind begrifflich so miteinander verkoppelt, daß der eine Begriff wesentlich durch den anderen definiert wird.
In this study,the chemical components of the Vinca rosea l. (Apocynaceae) leaves in water and alcoholic extracts were glycosides , phenols , tannins , resins , alkaloids and saponins , the medium of two extracts were acidic. The effect of theses extracts on the different microorganisms were studied. It has been found that (1mg/ml) concentration was effective on the inhibition of the growth of the intended microorganisms, and the inhibition of the hyaluronidase enzyme by these extracts were (25%) ,(27.9%) ,respectively. There were concentrations of the following trace elements in the leaves (K,Ca,Fe,Zn) with (227.2 , 33.2 , 26.4 , 17.3) ppm respectively.
The vascular permeability-increasing action of rabbit PMNL lysosomes has been studied in skin and cremaster muscle of the rat.
Both an extract of frozen-thawed granules and a cathepsin-free cationic protein fraction of the granules (which had previously been demonstrated to cause leukocyte adhesion and emigration in vivo) induce increased vascular permeability in skin and muscle which resembles that produced by histamine or histamine-liberators with respect to the timing of the response and the predominant type of microvessel affected.
Extracts of frozen-thawed lysosomes and the inflammatory lysosomal cationic protein both cause disruption of rat mesenteric mast cells in vitro, whereas a granule-free cytoplasmic fraction of PMN leukocytes and a non-inflammatory cationic protein fraction of the granules do not do so under identical test conditions. The mastocytolytic action of lysosomal materials in vitro is not inhibited in the presence of 10 kallikrein-inhibiting units of trasylol per ml. The mast cell rupturing fraction of PMNL granules (cationic protein) possesses no detectable peroxidase activity or acid-mucopolysaccharase activity. When compared with compound 48/80 on the basis of estimated molecular weight, the lysosomal cationic protein appears to be at least as active as the latter compound with respect to in vitro mastocytolytic potency.
Chronic pretreatment of rats with an agent known to reduce tissue mast cell numbers causes marked suppression of the vascular permeability change normally induced in skin and muscle by lysosomal extracts and cationic protein. Similar results are obtained if lysosomal materials are tested in rats pretreated with an antihistaminic.
These observations are discussed with respect to the mode of action of PMNL lysosomes in the early and late phases of local tissue-injury reactions.
Hyaluronic acid is an evolutionarily ancient molecule commonly found in vertebrate tissues and capsules of some bacteria. Here we review modern data regarding structure, properties, and biological functions of hyaluronic acid in mammals and Streptococcus spp. bacteria. Various aspects of biogenesis and degradation of hyaluronic acid are discussed, biosynthesis and degradation metabolic pathways for glycosaminoglycan together with involved enzymes are described, and vertebrate and bacterial hyaluronan synthase genes are characterized. Special attention is given to the mechanisms underlying the biological action of hyaluronic acid as well as the interaction between polysaccharide and various proteins. In addition, all known signaling pathways involving hyaluronic acid are outlined. Impaired hyaluronic acid metabolism, changes in biopolymer molecular weight, hyaluronidase activity, and enzyme isoforms often accompany carcinogenesis. The interaction between cells and hyaluronic acid from extracellular matrix that may be important during malignant change is discussed. An expected role for high molecular weight hyaluronic acid in resistance of naked mole rat to oncologic diseases and the protective role of hyaluronic acid in bacteria are discussed.
Unsere Vorstellungen von der Entzündung sind von jeher ein Abbild unserer biologischen Kenntnisse gewesen. Während im Altertum und Mittelalter das der äußeren Betrachtung zugängliche Syndrom der mit Erwärmung und Schmerzhaftigkeit verbundenen Rötung und Schwellung (rubor et tumor cum calore et dolore)3 für das Wesen der Entzündung gehalten wurde und man in der Neuzeit ihren Kernpunkt zunächst noch in der den Kardinalsymptomen zugrunde liegenden örtlichen Kreislaufstörung suchte, ist es in den letzten hundert Jahren mit der Zunahme unseres Wissens immer deutlicher geworden, daß es bei der Entzündung überhaupt kein „Hauptcharakteristikum“4 gibt. Vielmehr handelt es sich um eine Kette bestimmt charakterisierter Vorgänge, deren einzelne Glieder durch verschieden starkes Hervortreten der Entzündung ihr jeweiliges Gepräge geben. Im Anfang dieser Kette steht eine durch eine Entzündungsursache bedingte Störung des physiologischen Gleichgewichts, am Ende eine Wiederherstellung dieses Gleichgewichts. Dazwischen liegen Reaktionen des Bindegewebsapparates, der darin enthaltenen Gefäße und Nerven und des in den Gefäßen befindlichen Blutes. Auch ist es immer deutlicher geworden, daß dieser Ausgleichsvorgang in vielen Fällen mit der Bildung von Antikörpern einhergeht, die Entzündung also ähnlich wie die als Stressreaktion5 bekannte Gleichgewichtsstörung häufig mit Anpassung verbunden ist. Auch hat sich gezeigt, daß hierbei neben örtlichen Vorgängen auf dem Entzündungsfelde Reaktionen der regionären Lymphknoten und selbst entfernter Strukturen eine erhebliche Rolle spielen. Eine moderne Lehre von der Entzündung darf sich daher nicht auf die örtliche, durch die Entzündungsursache bedingte Störung und ihre Ausgleichung beschränken, sondern muß auch die damit verbundenen Anpassungsvorgänge und die entfernten Reaktionen mit in Betracht ziehen.
Das rheumatische Fieber stellt eine entzündliche Systemerkrankung des Bindegewebes dar, dessen ubiquitäres Vorkommen den vielfältigen Organbefall im Verlauf der Erkrankung verständlich macht. Die klinische Bedeutung des interindividuell wechselnden Organbefalls hat in der Nomenklatur bisher keinen allgemein anerkannten Niederschlag gefunden. So war in Deutschland die Bezeichnung „akute Polyarthritis“ für das Gesamtbild akuter rheumatischer Erscheinungen bis vor kurzem üblich. Erst in den jüngsten Publikationen findet sich die Bezeichnung „rheumatisches Fieber“ in Analogie zum anglo-amerikanischen „rheumatic fever“. Auch diese Bezeichnung ist nicht ganz befriedigend, da in der Anamnese zahlreicher eindeutiger rheumatischer Herzfehler ein Fieber nicht zu eruieren ist, jedoch berücksichtigt der erneut vorgeschlagene Name „rheumatische Infektion“ zu wenig den allergischen Faktor in der Pathogenese dieser Erkrankung, deren Initiator ohne Zweifel ein A-Streptokokken-Infekt ist. Die weltweite Verbreitung des Begriffes „rheumatic fever“ läßt es unzweckmäßig erscheinen, nach anderen Bezeichnungsweisen zu suchen.
A study of the fractions of the venom of Naja naja atra obtained by chromatography on sulphoethyl-Sephadex The venom of Naja naja atra was fractionated on sulphoethyl-Sephadex into fourteen clearly different fractions. The experimental conditions are described. A number of enzymatic activities were studied, viz. phospholipase A, cholinesterase, Lamino acid oxidase, adenosine triphosphatase, 5'-nucleotidase, ribonuclease, phosphodiesterase, nicotinamide dinucleotide phosphatase and hyaluronidase, as well as several inhibiting activities of enzymatic processes, viz. inhibition of anaerobic glycolysis, inhibition of the cytochrome oxidase system and inhibition of acetylcholinesterase. The following biological activities are described: toxicity, anticoagulation, cytotoxicity, and the direct haemolytic factor. The well separated fractions were analysed by electrophoresis on acrylamide gel and their molecular size was estimated by means of gel filtration.RésuméLe venin de Naja naja atra a été fractionné en quatorze fractions bien distinctes par chromatographie sur sulphoéthyl-Sephadex. La composition de ces fractions est étudiée. Un certain nombre d'activités enzymatiques ont été mesurées: la phospholipase A, la cholinestérase, la l-amino acide oxydase, l'adénosine triphosphatase, la 5'-nucléotidase, la ribonucléase, la phosphodiestérase, la nicotinamide dinucléotide pyrophosphatase et l'hyaluronidase. Plusieurs inhibiteurs de processus enzymatiques ont été estimés (inhibition de la glycolyse anaérobie, inhibition du systéme cytochrome oxydase et inhibition de l'acétylcholinestérase). Certaines activités biologiques, comme la toxicité, le pouvoir anticoagulant, la cytotoxicité, le facteur lytique direct sont également décrites. Les électrophérogrammes en gel d'acrylamide sont donnés pour les fractions les mieux séparées; les répartitions de tailles moléculaires dans ces fractions ont été estimées par la technique de la chromatographie sur gel de dextrane.
Wie bereits im letzten Kapitel erwähnt wurde, ist die Abgrenzung der Oligosaccharidasen von den Polysaccharidasen verhältnismäßig einfach; die Unterscheidung ist durch die Kettenlänge der Substrate gegeben.
Im Folgenden sollen der bisherigen Gepflogenheit folgend, unter Präzipitation die Ausfällung einer ursprünglich (molekular- oder kolloidispers) gelösten Substanz und unter Agglutination die Zusammenballung und sich daran anschließende Sedimentierung von zelligen Antigenen (Bakterien, Blutzellen [Hämagglutination] u. a.) verstanden und soweit es der Zusammenhang erlaubt, getrennt behandelt werden. Da der Vorgang der Zusammenballung und Ausflockung bekanntlich in kolloiden Lösungen sowie in gröber dispersen Systemen durch physikalisch-chemische Einflüsse allein ohne Einwirkung spezifischer Immunkörperreaktionen zustandekommen kann, so muß man die Mitwirkung solcher Vorgänge auch bei der spezifischen Präzipitation und Agglutination in Betracht ziehen, so daß es nötig ist, in kurzen Zügen die Faktoren zu erörtern, die für die Stabilität wie auch für die unspezifische Ausfällung von dispersen Systemen verantwortlich sind.
Willoughby, Donald S. (University of Minnesota, Minneapolis), and Dennis W. Watson. Host-parasite relationships among group A streptococci. II. Influence of sex on the susceptibility of inbred mice toward streptococcal infection. J. Bacteriol.
87
1457–1461. 1964—BALB/Sy mice showed marked sex differences in susceptibility to a strain of Streptococcus pyogenes type 18. Castration of the highly susceptible normal adult male increased their resistance 1,000-fold, which approximated that of normal female mice. Treatment of male mice with estrogen and stilbestrol did not affect their resistance significantly. Young, sexually immature male mice, 4 weeks old, were more resistant than mature males. Susceptibility of the resistant females was not affected by age or bilateral ovariectomy, but testosterone injections caused an increase in susceptibility. Treatment of male mice with estrogen and stilbestrol did not modify their resistance. Smears of peritoneal exudates from normal male mice after intraperitoneal injection of streptococci revealed rapid multiplication of the organisms. There was no evidence of multiplication in the resistant normal female and castrated male mice.
Pembangunan ubat sirap batuk dari sirap nanas dengan campuran poli herba. Mengkaji ciri-ciri kimia, rasa dan anti radang.
Regarding the pathogens that cause intractable acute otitis media in adults, mucoid type Streptococcus pneumoniae, which forms mucoid type colonies on blood agar plates, is well known, and the occurrence of otitis media due to this bacterial species is called mucosus otitis media. We encountered an adult patient with intractable acute otitis media due to Group A Streptococcus (GAS) who showed clinical characteristics that were similar to those observed in mucosus otitis media. A 55-year-old female visited the Eura ENT Clinic due to pharyngeal pain and left otorrhea. Left external acoustic meatus swelling, left serous otorrhea, and left mixed hearing loss were observed. A diagnosis of acute otitis media due to GAS was made by a rapid GAS antigen detection test using the otorrhea, and treatment with a series of antibiotics (garenoxacin, ceftriaxone, amoxicillin, and minocycline) was performed, but this regimen could not resolve the left otorrhea. She was therefore admitted in Nishinihon Hospital and received the drip intravenous infusion of piperacillin and a steroid. On the second hospitalization day, the otorrhea was resolved, and she thereafter steadily improved. In adults presenting with intractable acute otitis media, not only mucosus otitis media, but also acute otitis media due to GAS should therefore be considered in the differential diagnosis.
Many bacterial extracellular polysaccharides, which play an important role in various biological processes, contain glucuronic acid (GlcA) as a major component. Glucuronosyltransferase (GlcA-T) is the enzyme responsible for GlcA attachment in the biosynthesis of these polysaccharides. GlcA-T has recently attracted significant attention because of its biological activities as well as its potential application to chemoenzymatic synthesis of GlcA-containing oligosaccharides and polysaccharides that are difficult to achieve chemically due to the problems related to glycosylation reactions with GlcA. At present, nine GlcA-Ts derived from Xanthomonas campestris, Pasteurella multocida, Escherichia coli, Sphingomonas paucimobilis, and Streptococci have been characterized. This article reviews the recent progresses made in understanding the structures, functions, physical, and chemical properties of reported GlcA-Ts.
Als Mucopolysaccharide werden alle hexosaminhaltigen Polysaccharide bezeichnet. Je nach Fehlen oder Vorkommen saurer Gruppen unterteilt man weiter in neutrale und saure Mucopolysaccharide. Sie können als solche frei vorkommen; wenn sie aber mehr oder weniger fest an Peptide bzw. Proteine zu hochmolekularen Komplexen gebunden sind, bedarf es zu ihrer Freisetzung mehr oder minder energischer Methoden. Ist die Bindung salzartig (natürlich nur bei sauren Mucopolysacchariden möglich), soll von Mucoproteiden, im Falle echter, kovalenter Bindung dagegen von Mucoiden gesprochen werden.
By turbidity method, the hyaluronidase activity of the tissue extract and the antihyaluronidase activity of the serum were compared between patients with gastric cancer and gastric ulcer and the following results were obtained:
1. HD-activity of the carcinomatous tissue was much higher than that of binign ulcerous tissue, and anti-HD-activity of the serum in cancer patients was also more increased than that in the patients of gastric ulcer.
2. In general, activities of HD and anti-HD showed the tendency to increase or decrease in parallel with extracellular fluid (Thiocyanate space).
3. HD-activity of the microscopically normal stomach tissue distant from the carcinomatous ulcer was decreased remarkably than that of the carcinomatous tissue.
4. On the carcinomatous ulcer itself. it was recognized that HD activity is little different according to the part or place of the ulcer.
5. The remarkable differences of HD-activity among Borrmann's types were not recognized, but type III and IV showed a little higher activity than that of type I and II.
6. As the result of experiments on dogs, it was confirmed that hyaluronidase increased extracellular fluid and the same action was recognized in the cancerous extract also.
7. In the experimental study on rabbits, it was recognized that hyaluronidase accelerated the diffussion's ability of the pigment in the intracutaneous tissue.
Als Mucopolysaccharide werden alle hexosaminhaltigen Polysaccharide bezeichnet. Je nach Fehlen oder Vorkommen saurer Gruppen unterteilt man weiter in neutrale und saure Mucopolysaccharide. Sie können als solche frei vorkommen; wenn sie aber mehr oder weniger fest an Peptide bzw. Proteine zu hochmolekularen Komplexen gebunden sind, bedarf es zu ihrer Freisetzung mehr oder minder energischer Methoden. Ist die Bindung salzartig (natürlich nur bei sauren Mucopolysacchariden möglich), soll von Mucoproteiden, im Falle echter, kovalenter Bindung dagegen von Mucoiden gesprochen werden.
Streptococcal diseases of man have occurred for millenia, but it was not until the 19th century that the association between the etiological agent and the various clinical forms of the disease was learned. The principal forms of streptococcal disease now recognized are streptococcal sore throat, scarlet fever, streptococcal skin infection (impetigo, or pyoderma, and erysipelas). Streptococci may also cause suppurative infections (abscesses, pneumonia), food poisoning, and systemic disease (septicemia, endocarditis). Of significant clinical and public-health importance are the nonsuppurative sequelae of streptococcal disease, namely, rheumatic fever and acute glomerulonephritis.
Die Behauptung Rickers (1924), daß man die Entzündung nicht definieren kann, ist sicher ebenso unhaltbar wie seine Angabe, daß jede teleologische Deutung als unwissenschaftlich abgelehnt werden muß. Wenn es bisher nicht gelungen ist, die Entzündung allgemein befriedigend zu definieren, wenn auch bei der letzten großen Aussprache über diesen Vorgang auf der 19. Tagung der Deutschen Pathologischen Gesellschaft im Jahre 1923 „fast jeder einzelne eine Neigung gezeigt hat, den Ausdruck ,entzündlich in seinem eigenen Sinne zu gebrauchen“, so besagt das doch nur, daß das Wissen um die Entzündung zu jener Zeit noch zu unvollständig war, um eine befriedigende Begriffsbestimmung zuzulassen. Wie wohl bekannt ist, war Ricker, ähnlich wie Bier (1933), mit den durch Metschnikoff (1880–1913) entdeckten cellulären Abbauvorgängen wenig vertraut. Auch wußte er noch nichts von den Anpassungsvorgängen, welche die durch Antigene verursachten Entzündungen begleiten. Wären ihm diese Kardinalvorgänge der Entzündung bekannt gewesen, hätte es ihm kaum entgehen können, daß dieser Vorgang einen defensiven Charakter hat.
Hyaluronidase has been investigated in various strains of pneumococci and hemolytic streptococci, and in some material of
animal origin. The enzyme activity was measured by a viscosimetric method using as a substrate a fluid containing hyaluronic
acid as the viscous component, and by the hydrolysis of pure hyaluronic acid into its reducing components.
In pneumococci the enzyme was demonstrated in all types and in all strains tested, including smooth and rough forms of Types
I, II, III, and VI.
In hemolytic streptococci the enzyme from strain H44, group A, reported previously, was further investigated. In this strain,
as well as in other hemolytic streptococci containing the enzyme, great variability of the enzyme concentration was found.
Furthermore, the enzyme proved to be very labile, giving in the viscosimetric measurements a typical stoppage of the activity
initially present. In 13 out of 14 other strains of group A organisms investigated, no enzyme was demonstrable, but the variation
in activity in the enzyme-active strains renders the negative findings inconclusive. A very active enzyme, though of great
variability, was found in one group C strain.
The enzyme was prepared from the leech in confirmation of the work of Claude.
The enzyme from testis showed a maximum at pH 4.4 in contrast to the optimum of 5.8 in pneumococcal, streptococcal, and Cl. welchii preparations. The pH curve of the testis enzyme indicated, however, a second optimum coinciding with that of the bacterial
enzymes. The hydrolysis further indicated a break at about 50 per cent hydrolysis, indicating primarily the hydrolysis down
to aldobionic acid units. The depolymerizing action of testis enzyme is more marked than that of pneumococcal enzyme. The
results have been interpreted as due to the presence of two enzymes, one attacking the long chain molecule, the other hydrolyzing
the aldobionic acid formed.
The enzyme was further prepared from beef spleen. Here the hydrolysis of ß-glucuronides was compared to that of hyaluronic
acid. The two actions apparently are catalyzed by two distinct enzymes.
Enzyme preparations were further obtained from rabbit skin. Since hyaluronic acid has also been found in the skin, this organ
may play a considerable rôle in the metabolism of hyaluronic acid.
In addition to hyaluronic acid, it has been shown that hyaluronidases also hydrolyze the sulfuric acid containing polysaccharide
of the cornea. This polysaccharide has previously been characterized as a natural sulfuric acid ester of hyaluronic acid.
The pneumococcal enzyme preparations also attacked a polysaccharide acid prepared from submaxillary gland, which is not hyaluronic
acid. However, it is believed that this hydrolysis is due to a second enzyme contained in the preparations. The testis enzyme,
on the other hand, attacked chondroitinsulfuric acid and also contained a sulfatase.
The depolymerizing action of hyaluronidase has been discussed. It is concluded that depolymerization and hydrolysis are probably
due to the same enzyme attacking hyaluronic acid. It is suggested that the first attack of the enzyme does not cause an opening
of glucosidic linkages. The available evidence indicates that the viscosity of the natural fluids is not due to macromolecules
but to micellae formation, and that these micellae are depolymerized by the enzymatic reaction. It is assumed that the depolymerization
is due to a primary enzyme-substrate reaction, which in itself is insufficient to open the glucosidic linkages. The latter
reaction involves further steps.
The relationship between hyaluronidase and "spreading factor" has been discussed anew. Though more data have been reported
pointing to the identity of hyaluronidase and "spreading factor," our inability to demonstrate hyaluronidase in streptococcal
material of high "spreading" potency, is still a serious obstacle to the unitarian theory. However, it seems possible that
the streptococcal material may contain reversibly inactive enzyme which may be reactivated in vivo.
1. A rapid method for the roughly quantitative estimation of mucoid polysaccharide in hemolytic streptococci has been described. 2. Using this method, about 94 per cent of strains from moderate or severe streptococcal infections in man have been found to produce mucoid polysaccharide in greater or less amount. In a group of streptococci from normal throats only about 8 per cent produced this substance, all of the producers falling into Lancefield's group A. 3. Of the Lancefield group A strains from both normal and infected sources, 92 per cent showed the presence of mucoid polysaccharide in culture dilutions of 1:10 or higher. 4. The probable significance of the mucoid polysaccharide in streptococcal virulence is indicated.
1. Confirming the observations of other experimenters, it has been found that group A hemolytic streptococci produce a capsule containing a polysaccharide which is similar to, if not identical with, certain high molecular weight sugars found in the mammalian body. 2. Leech extract possesses a powerful enzyme capable of splitting one of the linkages in this polysaccharide and of decapsulating group A and group C hemolytic streptococci in vitro and in vivo. 3. Mice and guinea pigs can be protected from intraperitoneal infection with a virulent group C streptococcus by the intraperitoneal administration of leech extract. In contrast there is little protective action of leech extract in mice infected with group A hemolytic streptococci. 4. The protective effect of leech extract against streptococcal group C infection is probably due to the removal of the capsule in vivo. 5. The capsule of mouse virulent group C streptococci plays a major rôle in the virulence of that microorganism, while the capsule of certain mouse virulent group A streptococci plays little, if any, rôle in virulence, at least when the infection is intraperitoneal in the mouse.
A comparative study of spreading factor and hyaluronidase in preparations from various sources revealed the following points of similarity and dissimilarity in the two reactions. 1. Similarities: (a) All preparations containing hyaluronidase also produced spreading. (b) Heating at 65 degrees and 100 degrees C. for 30 minutes produced a comparable effect on both reactions. (c) The demonstration of the presence of hyaluronic acid in skin offers a plausible explanation for the mechanism of spreading on the basis of hyaluronidase activity. 2. Dissimilarities: (a) No parallelism was observed in the degree of activity of spreading factor and hyaluronidase in the same preparations. (b) All preparations which produced spreading did not contain hyaluronidase. (c) Antisera to hyaluronidase preparations specifically and completely inhibited the activity of the homologous enzyme but did not inhibit the spreading factor in the same preparations. The significance of the similarities and dissimilarities between the two reactions is discussed. It is concluded that while hyaluronidase may play a rôle in the spreading reaction the phenomenon is a complex one and cannot be explained on the basis of a simple chemical reaction.
1. Two qualitatively different type-specific antigens, designated M and T, have been found present in matt variants of group A hemolytic streptococci, but only one of these, the T antigen, occurs in the degraded glossy variant. 2. The protein nature of the M antigen, present in matt variants only, has been demonstrated in previous work, but the chemical characteristics of the newly recognized antigenic factor, T, present in both variants, have not been determined. This T factor is identified only by its immunological reactions. It is unknown whether the two type-specific antigenic factors, M and T, occur as separate chemical entities in the matt variant or in conjugation. 3. Antibody to the type-specific protein, M, appears responsible for the M precipitin reaction, for type-specific protection, and, as a rule, for part of the type-specific agglutination of matt variants, but in type 1 it does not cause agglutination. 4. Antibody to the second type-specific antigen, T, seems to be solely responsible for type-specific agglutination of the glossy form and to play a large rôle in type-specific agglutination of the matt form, but apparently it is not involved in protection. This T antibody causes all of the type-specific agglutination of type 1. Consequently, type 1 matt and glossy variants agglutinate and absorb agglutinin alike, and antisera to both are identical in content of type-specific agglutinin though they differ in respect to M antibody. 5. Recognition of the principle underlying type-specific agglutination of glossy variants makes it possible to suggest, with certain reservations, the use of glossy variants for type classification by agglutination. These variants are suitable for preparing type-specific agglutinating antisera, and they form stable suspensions for use in the reaction. Improved methods are needed for deriving glossy from matt variants.
An improved method is described for preparing the enzyme which hydrolyzes the polysaccharide acid contained in vitreous humor, umbilical cord, synovial fluid, and the mucoid phase of group A hemolytic streptococci. Preparations have been obtained from pneumococci, group A hemolytic streptococci, Clostridium welchii, and from splenic tissue, which display the same specific activity. Evidence is presented to show that the hydrolytic enzyme is not the same as that responsible for the lysis of pneumococci. In pneumococci and hemolytic streptococci the major portion of the enzyme is bound to the cell structure. The enzyme from Clostridium welchii is associated with other carbohydrate-splitting enzymes in the culture medium and not with the bacterial cells. It is suggested that the disappearance of the mucoid capsule of group A hemolytic streptococci is due to enzymatic hydrolysis of the acid polysaccharide. The relation between enzyme activity and the virulence and invasiveness of group A hemolytic streptococci is briefly considered.
A non-antigenic mucoid polysaccharide similar to that described by Kendall, Heidelberger, and Dawson was isolated from group C hemolytic streptococci. A simple method for its quantitative estimation is described. By means of this quantitative method, as well as by the direct isolation of the carbohydrates, the size and persistence of capsules in young cultures of various strains have been related to the non-antigenic mucoid polysaccharide. Similarly, invasiveness in different strains has been related to the mucoid polysaccharide. Data indicating the non-haptene nature of this material are presented. An autolytic process, accelerated by bile, is involved in the loss of capsular material from young streptococci.
The venom of several species of poisonous snakes acts to spread India ink through the skin as do the spreading factors procurable from certain tissues and elaborated by invasive bacteria. The factor is most abundant in the venom of the Viperidae (rattlesnake) family and relatively scant in the venom of Colubridae proteroglypha (cobra) family, and it is absent from toad venom. Extracts of the supralabial glands of harmless snakes contain only negligible amounts of the factor. Rattlesnake venom heated at 65 degrees to 100 degrees loses a large proportion of its toxicity but retains the ability to spread ink. Rattlesnake venom that has lost its toxicity on standing or on heating markedly enhances the infection produced by bacterial or virus suspension in the rabbit skin. Antivenine serum inactivates both the toxic and spreading factors of venom.
1. Anantiserum which specifically protects mice against a virulent culture (M variant) of the hemolytic streptococcus contains specific opsonin. Phagocytosis of the organisms can be observed in the peritoneum of the protected mouse. 2. An antiserum prepared by injecting an animal with the living M variant specifically opsonizes both the F and the M variant of the strain. 3. Evidence is presented which indicates the probable identity of the specific opsonin and the anti-M precipitin of Lancefield (7). Agglutination appears to be dependent upon a different antibody. 4. It is possible to type the hemolytic streptococci by means of specific opsonins, and the opsonic method has certain advantages over agglutination, precipitation, and mouse protection tests. It is evident from what little has been done that there are many types. 5. The serum of infants contains no opsonin for the virulent hemolytic streptococci, but the serum of adults may contain specific opsonins for certain strains. Inasmuch as no opsonins were demonstrable in two polyvalent antibacterial sera examined, the possibilities of therapeutic transfusion are discussed.
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