Molecular Cloning and Docking of speB Gene Encoding Cysteine Protease With Antibiotic Interaction in Streptococcus pyogenes NBMKU12 From the Clinical Isolates

Streptococcus pyogenes causes a variety of diseases ranging from mild diseases to severe invasive infections which result in significant morbidity and mortality. This study focuses on the antibiotic resistance of S. pyogenes and their interaction with cysteine protease. Around 36 beta-hemolytic isolates were collected from the clinical lab, of which seven isolates (19.4%) were identified as Streptococcus pyogenes. One of the seven isolates was collected from a urinary tract infection, which was identified by antibody agglutination and MALTI-TOF-MS, and it is designated as S. pyogenes NBMKU12. Around 8.3 to 66.6 % of the isolates were found to be resistant to one or more antimicrobial agents, especially, penicillin-G resistance was exhibited by 29.1% of the isolates. In the NBMKU12 isolate, the beta lactem (TEM) gene was detected among the 13 antibiotic genes for which it was tested. Furthermore, when analysis for presence of 13 virulence genes were carried out in NBMKU12 isolate, only speJ and speB were detected. The speB (streptococcal pyrogenic exotoxin B) encoding cysteine protease gene was cloned. This was followed by performing DNA sequencing to understand the putative cysteine protease interaction with antibiotics, inhibitors, and substrate. The speB gene consists of 1197 nucleotides and encodes a protein with multiple domains, including a signal peptide (aa 1–22), an inhibitor region (aa 27–156), and a catalytic cysteine domain (aa 160–367). The signal peptide cleavage site is predicted between Ala22 and Asn23. The putative 398 amino acid residues were found to have a theoretical pI of 8.76 and a molecular mass of 43,204.36 Da. The tested culture supernatants of NBMKU12 isolate exhibited the proteolytic activity against casein, papaya and pineapple used as substrates. The proteolytic activity suggests the expression of speB gene. Molecular docking analysis of cysteine protease showed that erythromycin (bond length 2.41 Å), followed by chloramphenicol (2.51 Å), exhibited a strong interaction; while penicillin-G (3.24 Å) exhibited a weak interaction, and this factor could be considered as a cause for penicillin-G resistance. The present study contributes to a better understanding of speB gene encoding cysteine protease, antibiotic resistance, and their interaction in the isolate, S. pyogenes NBMKU12. The antibiotics and cysteine protease interaction study confirms the resistance or sensitivity of S. pyogenes. Hence, it could be hypothesized that the isolate NBMKU12 is resistant to most of the tested antibiotics, and this resistance might be a cause for mutation

Phytochemical Screening, Antimicrobial Potential and GC-MS Analysis of Chloroform Leaf Extract of Hyptis suaveolens (L.) Poit

Bioactive compounds possessing therapeutic value are used for long time by vast group of researchers to treat various diseases for the goodness of the mankind. Most of the commercially available bioactive compounds are derived from plants because of their affirmative property on human health with minimum side effects. Hyptis suaveolens (L.) Poit. is one of the most important traditional and therapeutic plant, which is belongs to Lamiaceae family. The Biochemical test of the leaf extract of Hyptis suaveolens using different solvent shows the presence of biologically significant compounds. Thereby, the Antimicrobial properties of leaf extract of H. suaveolens carried out against six different bacterial pathogens and three fungi pathogens.  Most potential activity was observed in the crude extract of Hexane, Ethyl acetate and Chloroform against E.coli, Enterococcus faecalis and Streptococcus pyrogenes respectively when compared to Klebshilla pneumonia. Among the other crude extract, the chloroform crude extract resulted with more number of molecules in the phytochemical analysis and also it exhibits superior antibacterial activity. Hence, the Gas Chromatography-Mass Spectrometry (GC-MS) analysis of chloroform extract was achieved and 17 different bioactive compounds were discovered.  Neophytadiene, (S,Z)-Heptadeca-1,9-dien-4,6-diyn-3-ol, 2,3-Diazabicyclo[2.2.1]hept-2-ene, 7,7-dime and  Stigmasterol are the significant molecules present in the chloroform leaf extract it has the highest value for the mankind

Protection against discolouration by two over-the counter desensitising products

To determine in vitro the protection potential against discolouration of two OTC (over-the-counter) desensitising products on enamel and dentin in comparison to a standard toothpaste and water by means of a spectrophotometer

Does age influence self-perception of the soft-tissue profile in children?

Introduction: Appreciation of the soft-tissue profile is important in orthodontic diagnosis and treatment. However, are the patients themselves aware of their profile appearance? We aimed to evaluate if age influences self-perception of the soft-tissue profile in children. Methods: The study population for this prospective cross-sectional investigation consisted of 3 groups of 60 patients, classified according to age (&lt;12 years; 12-15 years; &gt;15 years). Each subject's right-sided facial profile was photographed to obtain a silhouette. Facial profile silhouette templates were created to represent the local population. Each subject's photograph was inserted into the corresponding template, and the subjects were asked to identify themselves. Facial profile self-recognition was recorded as a binary variable (yes or no). Other recorded variables included age, sex, and sexual maturity rating (using Tanner staging). Chi-square tests were used to analyze facial profile self-recognition between different subgroups, and stepwise multiple regression was used to predict the probabilities of facial profile self-recognition, with age, sexual maturity rating, and other recorded variables as independent variables. Results: Eighty percent of subjects aged &gt;15 years recognized their own profile, compared with only 55% and 50% of subjects aged 12-15 years and &lt;12 years, respectively. Subjects aged &gt;15 years were significantly more likely to recognize their profile than younger subjects (P = 0.001). Similarly, subjects with the most advanced sexual maturity rating (stage V) were significantly more likely to recognize their profile (85% self-recognition) than those in groups I-IV (P &lt;0.001). Girls were more likely to recognize their profiles than boys (P = 0.028). When using multiple regression analysis, sexual maturity rating appears to be the only significant predictor for facial profile self-recognition (R2 = 0.25; P &lt;0.001). Conclusions: Facial profile self-recognition seems to improve with age and sexual maturity (sexual maturity rating stage V). Because orthodontic treatment planning takes possible soft-tissue changes into account, it is important to evaluate the degree of self-perception of the patients to adapt our goals and treatment discussions.</p

Biocontrol Properties and Functional Characterization of Rice Rhizobacterium Pseudomonas sp. VSMKU4036

A total of 30 fluorescent pseudomonads (FPs) were showed significant antagonistic activity against different fungal phytopathogens with different level of the zone of inhibition (ZOI) for Rhizoctonia solani (5mm-34mm), Macrophomina phaseolina (9mm-37mm), Scleotium rolfsii (4mm-36mm), Helminthusporium solani (5mm-27mm), Fusarium oxysporum (2mm-25mm) and Fusarium oxysporum RACE (4mm-31mm) compared to control. The maximum growth of our selected isolate VSMKU4036 was observed in King’B Broth (KBB), pH 7.0 and at 37°C. The VSMKU4036 isolate has been recognized as Pseudomonas sp, based on the morphological, biological, and different functional characteristics. Antagonistic rhizobacterium Pseudomonas sp VSMKU4036 produced antimicrobial traits, such as plant growth promotion and various functional characters like siderophores, hydrogen cyanide (HCN), phosphate solubilization, indole acetic acid (IAA), biofilms formation, protease, gelatinase, amylase, and pectinase. Our superior biocontrol isolate VSMKU4036 was high resistance to tetracycline, streptomycin and nalidixic acid, however, it was sensitive to ampicillin and rifamycin. Pseudomonas sp VSMKU4036 showed maximum resistance to cadmium, nickel chloride, copper sulphate, magnesium sulphate, zinc chloride and ferric chloride where as highly sensitive to mercuric chloride, and selenium dioxide compared to control

Table_1_Molecular Cloning and Docking of speB Gene Encoding Cysteine Protease With Antibiotic Interaction in Streptococcus pyogenes NBMKU12 From the Clinical Isolates.DOCX

<p>Streptococcus pyogenes causes a variety of diseases ranging from mild diseases to severe invasive infections which result in significant morbidity and mortality. This study focuses on the antibiotic resistance of S. pyogenes and their interaction with cysteine protease. Around 36 beta-hemolytic isolates were collected from the clinical lab, of which seven isolates (19.4%) were identified as Streptococcus pyogenes. One of the seven isolates was collected from a urinary tract infection, which was identified by antibody agglutination and MALTI-TOF-MS, and it is designated as S. pyogenes NBMKU12. Around 8.3 to 66.6 % of the isolates were found to be resistant to one or more antimicrobial agents, especially, penicillin-G resistance was exhibited by 29.1% of the isolates. In the NBMKU12 isolate, the beta lactem (TEM) gene was detected among the 13 antibiotic genes for which it was tested. Furthermore, when analysis for presence of 13 virulence genes were carried out in NBMKU12 isolate, only speJ and speB were detected. The speB (streptococcal pyrogenic exotoxin B) encoding cysteine protease gene was cloned. This was followed by performing DNA sequencing to understand the putative cysteine protease interaction with antibiotics, inhibitors, and substrate. The speB gene consists of 1197 nucleotides and encodes a protein with multiple domains, including a signal peptide (aa 1–22), an inhibitor region (aa 27–156), and a catalytic cysteine domain (aa 160–367). The signal peptide cleavage site is predicted between Ala22 and Asn23. The putative 398 amino acid residues were found to have a theoretical pI of 8.76 and a molecular mass of 43,204.36 Da. The tested culture supernatants of NBMKU12 isolate exhibited the proteolytic activity against casein, papaya and pineapple used as substrates. The proteolytic activity suggests the expression of speB gene. Molecular docking analysis of cysteine protease showed that erythromycin (bond length 2.41 Å), followed by chloramphenicol (2.51 Å), exhibited a strong interaction; while penicillin-G (3.24 Å) exhibited a weak interaction, and this factor could be considered as a cause for penicillin-G resistance. The present study contributes to a better understanding of speB gene encoding cysteine protease, antibiotic resistance, and their interaction in the isolate, S. pyogenes NBMKU12. The antibiotics and cysteine protease interaction study confirms the resistance or sensitivity of S. pyogenes. Hence, it could be hypothesized that the isolate NBMKU12 is resistant to most of the tested antibiotics, and this resistance might be a cause for mutation.</p