Bacterial Deoxyribonucleic Acid Research Articles

Bacterial dysbiosis in newly diagnosed treatment naïve pediatric ulcerative colitis in Saudi Arabia.

The role of microbiota in the pathogenesis of ulcerative colitis (UC) has been increasingly recognized. However, most of the reports are from Western populations. In Middle Eastern countries, including Saudi Arabia, little is known about the role of microbiota. Therefore, our aim was to describe the bacterial microbiota profile and signature in pediatric UC in Saudi Arabia. Twenty children with UC and 20 healthy controls enrolled in the study gave stool samples. Twenty rectal mucosal samples were taken from UC and 20 from non-UC controls. Inclusion criteria included newly diagnosed and untreated children and lack of antibiotic exposure for at least 6 months before stool collection was required for children with UC and controls. Bacterial deoxyribonucleic acid was extracted and sequenced using shotgun metagenomic analysis. Statistical analysis included Shannon alpha diversity metrics, Bray-Curtis dissimilarity, DESeq2, and biomarker discovery. The demographic characteristics were similar in children with UC and controls. There was a significant reduction in alpha diversity (P = 0.037) and beta diversity in samples from children with UC (P = 0.001). Many taxa were identified with log2 abundance analysis, revealing 110 and 102 species significantly depleted and enriched in UC, respectively. Eleven bacterial species' signatures were identified. In Saudi Arabian children with UC, we demonstrate a dysbiosis similar to reports from Western populations, possibly related to changes of lifestyle. Microbial signature discovery in this report is an important contribution to research, leading to the development of adjunctive non-invasive diagnostic options in unusual cases of UC.

Analysis of gut microbiotal diversity in healthy young adults in Sunan County, Gansu Province, China.

To examine gut microbiotal diversity in the Han Chinese and Yugur populations of Sunan County, Gansu Province, living in the same environmental conditions, and to analyze possible causes of differences in diversity. We selected 28 people, ages 18-45 years old, all of whom were third-generation pure Yugur or Han Chinese from Sunan County. Fresh fecal samples were collected, and total bacterial deoxyribonucleic acid (DNA) was extracted. We performed 16S ribosomal ribonucleic acid (16S rRNA) high-throughput sequencing (HTS) and bioinformatics to study the relationships among between gut microbiota structure, genetics, and dietary habits in Yugur and Han Chinese subjects. We found 350 differential operational taxonomic units (OTUs) in Han Chinese and Yugur gut microbiota, proving that gut microbiota differed between the two populations. That were less abundant among Yugurs than Han Chinese were Prevotella_9 and Alloprevotella. That were more abundant among Yugurs than Han Chinese were Anaerostipes and Christensenellaceae_R-7_group. And they were significantly associated with a high-calorie diet In addition. we found differences in predicted gut microbiota structural functions (The main functions were metabolic and genetic information) between the two populations. Yugur subjects demonstrated differences in gut microbiotal structure from Han Chinese subjects, and this difference influenced by dietary and may be influenced by genetic influences. This finding will provide a fundamental basis for further study of the relationships among gut microbiota, dietary factors, and disease in Sunan County.

Design of Two Phase III, Randomized, Multicenter Studies Comparing Gepotidacin with Nitrofurantoin for the Treatment of Uncomplicated Urinary Tract Infection in Female Participants.

BackgroundUncomplicated urinary tract infections (uUTIs) are among the most common community-acquired infections for women worldwide. Treatment options are increasingly limited by antibiotic resistance; novel oral antibiotics are urgently needed. Gepotidacin is a novel, bactericidal, first-in-class triazaacenaphthylene antibiotic that inhibits bacterial deoxyribonucleic acid (DNA) replication by a distinct mechanism of action, which confers activity against most strains of target pathogens, such as Escherichia coli and Staphylococcus saprophyticus, including those resistant to current antibiotics. Here, we describe the designs of two near-identical phase III clinical trials (EAGLE-2 and EAGLE-3) evaluating gepotidacin for the treatment of uUTI.MethodsThese are phase III, randomized, multicenter, parallel-group, double-blind, double-dummy, comparator-controlled, noninferiority studies, comparing the efficacy and safety of gepotidacin to nitrofurantoin in the treatment of uUTI. Eligible participants are women aged ≥ 12 years with ≥ 2 uUTI symptoms, randomized (1:1) to receive oral gepotidacin (1500 mg) plus placebo or nitrofurantoin (100 mg) plus placebo, twice daily for 5 days. The primary therapeutic endpoint is composite clinical and microbiological efficacy, with noninferiority comparisons made in individuals with a qualifying (≥ 105 colony-forming units/mL urine) nitrofurantoin-susceptible uropathogen.ResultsThese trials were designed in accordance with US Food and Drug Administration (2019) and European Medicines Agency (2018) guidance, particularly the composite endpoint and microbiological evaluability requirements. Across the trials ~ 5000 participants are planned to be enrolled from > 200 centers globally.ConclusionsGepotidacin represents an important potential treatment option being evaluated to address the need for novel oral antibiotics to treat uUTI.These trials are registered at ClinicalTrials.gov (https://clinicaltrials.gov/) where the full protocols can be accessed under trial IDs: NCT04020341 (EAGLE-2) and NCT04187144 (EAGLE-3).Supplementary InformationThe online version contains supplementary material available at 10.1007/s40121-022-00706-9.

The Potential Role of Urinary Microbiome in Benign Prostate Hyperplasia/Lower Urinary Tract Symptoms.

Historically, urine in the urinary tract was considered “sterile” based primarily on culture-dependent methods of bacterial detection. Rapidly developing sequencing methods and analytical techniques have detected bacterial deoxyribonucleic acid and live bacteria in urine, improving our ability to understand the urinary tract microbiome. Recently, many studies have revealed evidence of a microbial presence in human urine in the absence of clinical infections. In women, fascinating evidence associates urinary tract microbiota with lower urinary tract symptoms (LUTS). However, the association between urinary tract microbiota and men with LUTS, particularly those with benign prostate hyperplasia (BPH), has not been established. In addition, the identification of the proinflammatory cytokines and pathogens responsible for the clinical progression of BPH is still underway. This review article aimed to address microbiome-related evidence for BPH. Further studies are required for a comprehensive understanding of the relationship between the urogenital microbiome and BPH pathogenesis to facilitate the development of preventive and therapeutic approaches for male LUTS.

Comparing protocols of DNA extraction from Escherichia coli: Analysis of purity and concentration by gel electrophoresis

Background and objective: Given the difficulty in establishing an ideal method that can successfully extract bacterial deoxyribonucleic acid (DNA) in Gram-negative bacteria, the objective of this work was to compare protocols for the extraction of bacterial DNA and to suggest more practical, faster, and less costly methodologies. Methods: Bacterial species used were Escherichia coli, provided by Dr. Leão Sampaio University Center. The evaluated/adapted methodologies were: sodium dodecyl sulfate (SDS); salting-out; Promega kit and cetyltrimethylammonium bromide (CTAB), and phenol/chloroform. Extracted DNA was examined by the agarose gel electrophoresis. Results: The SDS method revealed the best result in extracting DNA from E. coli. Its advantages include rapidness, low cost, and good concentration of the extracted material. Others methods showed low-quality DNA, probably due to the presence of large amounts of proteins in the cell wall of E. coli, interfering with the quality of the samples. Conclusions: It was concluded that the SDS method is better than others with better DNA quality, low cost, and good efficiency.

Surgical Helmets Used During Total Joint Arthroplasty Harbor Common Pathogens: A Cautionary Note

BackgroundThe use of personal-protection surgical helmet/hood systems is now a part of the standard surgical attire during arthroplasty in North America. There are no protocols for the disinfection of these helmets. MethodsThis is a prospective, single-center, observational study. Helmets worn by 44 members of the surgical team and foreheads of 44 corresponding surgical personnel were swabbed at three distinct time points. In addition, 16 helmets were treated with hypochlorite spray to determine if pathogens could be eliminated. Swabs obtained were processed for culture and next-generation sequencing (NGS). ResultsOf the 132 helmet samples, 97 (73%) yielded bacteria on culture and 94 (71%) had evidence of bacterial–deoxyribonucleic acid (DNA) on NGS. Of the swabs sent for bacterial identification at the three time points, at least one from each helmet was positive for a pathogen(s). Of the 132 forehead samples, 124 (93%) yielded bacteria on culture and 103 (78%) had evidence of bacterial-DNA on NGS. The most commonly identified organism from helmets was Cutibacterium acnes (86/132) on NGS and Staphylococcus epidermidis (47/132) on culture. The most commonly identified organism from the foreheads of surgical personnel was Cutibacterium acnes (100/132) on NGS and Staphylococcus epidermidis (70/132) on culture. Sanitization of helmets was totally effective; no swabs taken the following morning for culture and NGS identified any bacteria. ConclusionThis study demonstrates that surgical helmets worn during orthopedic procedures are contaminated with common pathogens that can potentially cause surgical site infections. The findings of this study should at the minimum compel us to develop protocols for the disinfection of these helmets.

Internet of things-based design of maternal and infant monitoring system and adoption of gold nanoparticles bacterial DNA detection technology in probiotic treatment of pregnancy reaction

it aimed to establish a maternal and child monitoring system based on medical Internet of Things (IoT) technology and explore the application of gold magnetic nanoparticles (Au-MNPs) in bacterial detection. The effect of active Bifidobacterium tablet (Siliankang) on intestinal flora of pregnant women and reaction to early pregnancy was analyzed. A real-time fetal heart rate monitoring method was established based on median filtering algorithm, and a maternal and infant monitoring system was established by organically combining portable monitoring equipment and hospital monitoring center through the IoT. Fe3O4 nanoparticles and Au-NPs were prepared by co-precipitation method and Turkevitch-Frens method. Fe3O4-Au-SiO2 NPs was obtained by polymerizing Au-NPs into Fe3O4 nanoparticles and coating the surface with SiO2. The characterization of NPs and its bacterial adsorption efficiency were analyzed, and a capture sensor for the detection of bacterial Deoxyribonucleic acid (DNA) was prepared. A total of 120 patients with pregnancy reaction were selected as the research object, and were divided into the placebo group and the experimental group (compound probiotics supplement group) according to different treatment methods, with 60 patients in each group. The bacterial DNA detection sensor was used to detect the difference of bacterial flora in feces of patients in different groups. The results showed that the particle size of Fe3O4-Au-SiO2 NPs was 15–47 nm, and the adsorption efficiency of Fe3O4-Au-SiO2 NPs to S. aureus, S. epidermidi, E. coli, and P. aeruginosa was higher than 80%. The Fe3O4-Au-SiO2 NPs of the prepared bacterial DNA detection sensor could be hybridized with the target bacterial DNA sequence. the intestinal enterobacteria, enterococci, and Bacteroides of patients in test group were remarkably reduced in contrast to control group, while the Bifidobacterium, Lactobacillus, and Fusobacterium flora were greatly increased (P < 0.05). the number of NVP patients in the control group and test group was 19 case (31.7%) and 5 case (8.3%). The probability of early pregnancy reaction was notably reduced (P < 0.05). It showed that the maternal and infant monitoring system based on Internet of things technology had potential application value for maternal and infant health. Tetralogy of viable Bifidobacterium tablets (Siliankang) can reduce the pathogenic bacteria in the intestines of pregnant women, and then reduced the incidence of early pregnancy reactions.

Screening of bacterial DNA in bile sampled from healthy dogs and dogs suffering from liver- or gallbladder-associated disease.

Although the biliary system is generally aseptic, gallbladder microbiota has been reported in humans and some animals apart from dogs. We screened and analyzed the bacterial deoxyribonucleic acid in canine gallbladders using bile sampled from 7 healthy dogs and 52 dogs with liver- or gallbladder-associated disease. PCR screening detected bacteria in 17.3% of diseased dogs (9/52) and none in healthy dogs. Microbiota analysis of PCR-positive samples showed that the microbial diversity differed between liver- and gallbladder-associated disease groups. Thus, a specific bacterial community appears to occur at a certain frequency in the bile of diseased dogs.

OPTIMIZATION OF GRAM POSITIVE BACTERIA DNA EXTRACTION PROTOCOLS AND PURITY AND CONCENTRATION ANALYSIS BY GEL ELECTROPHORESIS

This work aims to optimize protocols for extraction of bacterial Deoxyribonucleic Acid (DNA) in order to develop more practical, fast and less costly methodologies, therefore bringing improvements to the development of research and applications in laboratory routine based on the difficulty of establishing an ideal method that can extract Gram-positive bacterial DNA with excellent quality. The material and the bacterial strain Staphylococcus aureus 15 were provided by the Leão Sampaio University Center, where tests for DNA extraction were also carried out. The tests used were Sodium Dodecyl Sulfate (SDS); salting-out; PROMEGA’s Kit and CTAB / Phenol / Chloroform and were analyzed using agarose gel electrophoresis. The best results were obtained through the use of PROMEGA's commercial kit, salting-out without the use of Proteinase K and CTAB / Phenol / Chloroform, with salting-out having greater advantages in relation to other methods because it brings speed, low cost, good concentration of extracted material and not be toxic. The molecular tests for the extraction of Staphyloccocus aureus DNA were successful in general, when comparing issues such as low cost, extracted concentration, sharpness of the bands in electrophoresis and practicality of the performance. The best method for a small laboratory routine or for future research would be the salting-out without proteinase K, thus concluding that it is possible to obtain an excellent DNA extraction with less complexity and low cost methods.DOI: http://dx.doi.org/10.16891/2317-434X.v9.e2.a2021.pp1133-1140a

OPTIMIZATION OF GRAM POSITIVE BACTERIA DNA EXTRACTION PROTOCOLS AND PURITY AND CONCENTRATION ANALYSIS BY GEL ELECTROPHORESIS

This work aims to optimize protocols for extraction of bacterial Deoxyribonucleic Acid (DNA) in order to develop more practical, fast and less costly methodologies, therefore bringing improvements to the development of research and applications in laboratory routine based on the difficulty of establishing an ideal method that can extract Gram-positive bacterial DNA with excellent quality. The material and the bacterial strain Staphylococcus aureus 15 were provided by the Leão Sampaio University Center, where tests for DNA extraction were also carried out. The tests used were Sodium Dodecyl Sulfate (SDS); salting-out ; PROMEGA’s Kit and CTAB / Phenol / Chloroform and were analyzed using agarose gel electrophoresis. The best results were obtained through the use of PROMEGA's commercial kit, salting-out without the use of Proteinase K and CTAB / Phenol / Chloroform, with salting-out having greater advantages in relation to other methods because it brings speed, low cost, good concentration of extracted material and not be toxic. The molecular tests for the extraction of Staphyloccocus aureus DNA were successful in general, when comparing issues such as low cost, extracted concentration, sharpness of the bands in electrophoresis and practicality of the performance. The best method for a small laboratory routine or for future research would be the salting-out without proteinase K, thus concluding that it is possible to obtain an excellent DNA extraction with less complexity and low cost methods. DOI: http://dx.doi.org/10.16891/2317-434X.v9.e2.a2021.pp1133-1140a

OPTIMIZATION OF GRAM POSITIVE BACTERIA DNA EXTRACTION PROTOCOLS AND PURITY AND CONCENTRATION ANALYSIS BY GEL ELECTROPHORESIS

This work aims to optimize protocols for extraction of bacterial Deoxyribonucleic Acid (DNA) in order to develop more practical, fast and less costly methodologies, therefore bringing improvements to the development of research and applications in laboratory routine based on the difficulty of establishing an ideal method that can extract Gram-positive bacterial DNA with excellent quality. The material and the bacterial strain Staphylococcus aureus 15 were provided by the Leao Sampaio University Center, where tests for DNA extraction were also carried out. The tests used were Sodium Dodecyl Sulfate (SDS); salting-out ; PROMEGA’s Kit and CTAB / Phenol / Chloroform and were analyzed using agarose gel electrophoresis. The best results were obtained through the use of PROMEGA's commercial kit, salting-out without the use of Proteinase K and CTAB / Phenol / Chloroform, with salting-out having greater advantages in relation to other methods because it brings speed, low cost, good concentration of extracted material and not be toxic. The molecular tests for the extraction of Staphyloccocus aureus DNA were successful in general, when comparing issues such as low cost, extracted concentration, sharpness of the bands in electrophoresis and practicality of the performance. The best method for a small laboratory routine or for future research would be the salting-out without proteinase K, thus concluding that it is possible to obtain an excellent DNA extraction with less complexity and low cost methods. DOI: http://dx.doi.org/10.16891/2317-434X.v9.e2.a2021.pp1133-1140a

Evaluation of Pipemidic Acid Derivatives for Potential Antimicrobial Activity Application: In silico Studies on Bioactivity

Background: Pipemidic acid is a broad-spectrum quinolone antibacterial agent for the treatment of chronic urinary tract infections against both gram-positive and gram-negative bacteria. Both quinolone and fluoroquinolone antibiotics have been useful in combating bacterial infections. However, patients suffer severe side effects when they stop taking the medication. The piperazinyl region of pipemidic acid is highly responsible for the side effects. Objective: The objective of this study is to design new compounds in which the piperazinyl region is masked by way of conjugation to benzoic acid derivatives. Methods: In silico studies were conducted using AutoDockTools software for ligand-protein docking. The docking scores were compared to the parent pipemidic acid docked to Bacterial DNA (deoxyribonucleic acid) gyrase and GABA (gamma-Aminobutyric acid) receptor from the PDB (Protein Data Bank) database. Sites of metabolism, biological activity, quantum chemical descriptors, and ADME (absorption, distribution, metabolism, and excretion) property predictions for each designed ligand were also evaluated. Results: The docking studies and biological activity predictions showed good anti-infective properties (ligand PAR03) whilst also suggesting a reduction in GABA receptor agonist activity. The performance of PAR03 correlates with its electronic properties showing electrophilic character (can generate Reactive Electrophilic Species (RES)). Conclusion: The results from this study indicate that modification of the piperazinyl region of pipemidic acid can be an effective way to improve the drug potency whilst also ensuring reduction of the associated side effects.

Development of Electrochemical DNA Biosensor for Equine Hindgut Acidosis Detection.

The pH drop in the hindgut of the horse is caused by lactic acid-producing bacteria which are abundant when a horse’s feeding regime is excessively carbohydrate rich. This drop in pH below six causes hindgut acidosis and may lead to laminitis. Lactic acid-producing bacteria Streptococcus equinus and Mitsuokella jalaludinii have been found to produce high amounts of L-lactate and D-lactate, respectively. Early detection of increased levels of these bacteria could allow the horse owner to tailor the horse’s diet to avoid hindgut acidosis and subsequent laminitis. Therefore, 16s ribosomal ribonucleic acid (rRNA) sequences were identified and modified to obtain target single stranded deoxyribonucleic acid (DNA) from these bacteria. Complementary single stranded DNAs were designed from the modified target sequences to form capture probes. Binding between capture probe and target single stranded deoxyribonucleic acid (ssDNA) in solution has been studied by gel electrophoresis. Among pairs of different capture probes and target single stranded DNA, hybridization of Streptococcus equinus capture probe 1 (SECP1) and Streptococcus equinus target 1 (SET1) was portrayed as gel electrophoresis. Adsorptive stripping voltammetry was utilized to study the binding of thiol modified SECP1 over gold on glass substrates and these studies showed a consistent binding signal of thiol modified SECP1 and their hybridization with SET1 over the gold working electrode. Cyclic voltammetry and electrochemical impedance spectroscopy were employed to examine the binding of thiol modified SECP1 on the gold working electrode and hybridization of thiol modified SECP1 with the target single stranded DNA. Both demonstrated the gold working electrode surface was modified with a capture probe layer and hybridization of the thiol bound ssDNA probe with target DNA was indicated. Therefore, the proposed electrochemical biosensor has the potential to be used for the detection of the non-synthetic bacterial DNA target responsible for equine hindgut acidosis.

No distinction in the gut microbiota between diarrhea predominant-irritable bowel syndrome and healthy subjects: matched case\u2013control study in Thailand

BackgroundAlteration in the gut microbiota has been proposed in irritable bowel syndrome (IBS) pathogenesis, especially in the diarrheal type (IBS-D). We conducted this study to evaluate the fecal microbiota in Thai IBS-D patients when compared with healthy subjects as well as to evaluate the effects of probiotics on changes in the gut microbiota correlated with symptoms.MethodsA matched case–control study was conducted on diagnosed IBS-D patients, based on the Rome IV criteria and healthy controls. Stool samples were collected in preservation tubes. Bacterial deoxyribonucleic acid extraction was performed and amplified. Next, 16S ribosomal ribonucleic acid genes sequencing was performed to identify the microbiome in both the groups. IBS-D patients were provided with a probiotic mixture that was rich in Lactobacillus acidophillus and Bifidobacterium bifidum over 8 weeks. Changes in the symptoms, stool characteristics, and fecal microbiota were evaluated and compared with the corresponding baseline values.ResultsTwenty IBS-D patients with 20 age and gender-matched controls were included in this study. The baseline characteristics were not significantly different between the groups, including the mode of birth and the history of breastfeeding in infancy. No significant difference was noted in the fecal microbiota between the IBS-D patients and controls. The IBS symptom severity scales (IBS-SSS) were not statistically different after probiotic prescription; although, the bowel movements, the sense of urgency to go to the toilet and passing of mucous stool had obviously decreased. No change was noted in the fecal microbiota after receiving the experimental probiotic, except for an increase in the proportion of B. bifidum.ConclusionAlteration in the gut microbiota composition was probably not the main pathogenic mechanism in the Thai IBS-D patients assessed in this study. However, modifying microbiomes with potentially protective bacteria seems to be a beneficial therapy.Thai Clinical trial registry: TCTR20191211006, Date of registration: 10 Dec 2019. Retrospectively registered, Clinical trial URL: www.clinicaltrials.in.th

Evidence for Differentiation of Colon Tissue Microbiota in Patients with and without Postoperative Hirschsprung's Associated Enterocolitis: A Pilot Study.

PurposeTo investigate the differences in the colon microbiota composition of Hirschsprung's disease (HSCR) patients with and without a history of postoperative Hirschsprung's associated enterocolitis (HAEC).MethodsColon tissue microbiota was characterized by bacterial deoxyribonucleic acid (DNA) extraction and 16S rDNA sequencing for taxonomic classification and comparison.ResultsThe sequence diversity richness within samples was significantly higher in samples from patients with a history of postoperative HAEC. We observed an increased relative abundance of the phyla Bacteroidetes, Firmicutes and Cyanobacteria in HAEC patients and Fusobacteria, Actinobacteria and Proteobacteria in HSCR patients and, an increased relative abundance of the genera Dolosigranulum, Roseouria and Streptococcus in HAEC patients and Propionibacterium and Delftia in HSCR patients.ConclusionOur findings provide evidence that the colon tissue microbiota composition is different in HSCR patients with and without postoperative HAEC.

Bacterial DNA is a prognostic factor for mortality in patients who recover from spontaneous bacterial peritonitis.

BackgroundSpontaneous bacterial peritonitis (SBP) is associated with a high mortality. The aim was to investigate whether bacterial deoxyribonucleic acid (bactDNA) could offer an accurate identification of pathogens and to explore its prognostic role during and early after an SBP episode.MethodsConsecutive patients with SBP (SBP-group) and patients with decompensated cirrhosis without SBP/bacterascites (control-group) were enrolled. Standard culture methodology was used to isolate and identify pathogens from blood and ascitic fluid (AF). The SeptiFast test was used to identify bactDNA directly from AF.ResultsFifty-five patients, median age 60 (interquartile range [IQR] 53-74), model-for-end-stage liver disease (MELD) score 18 (IQR 13-29), with SBP were prospectively included. AF cultures were positive in 52.7% (17.2% drug-resistant bacteria) and bactDNA in 29.1% (58.2% combined sensitivity). BactDNA results were 84.6% concordant with AF cultures. Three patients had positive bactDNA in the culture-negative SBP-group. BactDNA was negative in all 36 of the control group (100% specificity). In multivariate analysis for 7-day survival, factors adversely affecting outcome were MELD (P=0.049) and C-reactive protein (P=0.012). After patients who died during the first week post-admission were excluded, patients with positive bactDNA had a poor prognosis compared to those with a negative test (log-rank P=0.005). Variables independently associated with 30-day mortality were neutrophil-to-lymphocyte ratio (P=0.011) and positive bactDNA (P=0.020).ConclusionsNo evidence was found for the usefulness of bactDNA to improve bacterial identification during an SBP episode. However, bactDNA was a predictor of 30-day mortality in the subset of patients who recovered from the infection episode.

Detection of blaCTX-M Genes among Extended Spectrum Beta Lactamase producing Pseudomonas aeruginosa isolated from Clinical Specimens in Erbil

Pseudomonas aeruginosa is one of the most common opportunistic pathogens that cause infections. The drug resistance in Pseudomonas aeruginosa may relate to the production of broad spectrum beta lactamase enzymes. This study aimed to detect multidrug resistant, extensively drug resistant and pan drug resistant isolates of Pseudomonas aeruginosa and to determine the frequency of extended spectrum beta lactamases enzymes by using a reliable test for detection blaCTX-M1, blaCTX-M2 and blaCTX-M3 genes in Pseudomonas aeruginosa. Pseudomonas aeruginosa isolated from different clinical samples, including sputum, wound, burn and urine. Identification and antibiotic susceptibility test were made by VITEK® 2 automated system. Identification confirmed by 16S ribosomal ribonucleic acid. The bacterial deoxyribonucleic acid was extracted and specific primers were used for detection blaCTX-M1, blaCTX-M2 and blaCTX-M3 genes. 62 Pseudomonas aeruginosa strains were obtained from 650 different clinical specimens (9.54 %). Of the 62 Pseudomonas aeruginosa isolates, the highest resistances were related to the antibiotics of ticarcillin, ticarcillin/clavulanic acid, piperacillin, piperacillin/tazobactam, ceftazidime and cefepime (100 %). However, the lowest resistance was observed to meropenem, imipenem (3.23 %), amikacin, ciprofloxacin (6.45 %) and gentamycin, tobramycin (19.35 %). Out of 62 bacterial isolates, 46 (74.19 %) were multidrugresistance strains, 2 (3.23 %) were extensive drug-resistance and no pan drug resistance was isolated. The gene frequency of blaCTX-M1 was 12 (19.35 %) and blaCTX-M2 5 (8.06 %), all specimens were negative for blaCTX-3 gene. This study highlights the increased prevalence of multidrug resistant Pseudomonas aeruginosa. The blaCTX-M genes provide Pseudomonas aeruginosa with an additional powerful resistance mechanism with potentially serious clinical implications, including limitation of the therapeutic options. The incidence of extended spectrum beta lactamases varies with geographic location and time.

Bacterial colonization of explanted non-endocarditis cardiac valves: evidence and characterization of the valvular microbiome.

Bacterial colonization has been already demonstrated in heart valve tissues of patients without cardiovascular infections. However, the evidence of a valvular microbiome is still scarce. The next-generation sequencing method was carried out on 34 specimens of aortic (n = 20) and mitral valves (n = 14) explanted from 34 patients having neither evidence nor history of infectious diseases, particularly infective endocarditis. While no bacteria were demonstrated using standard culture methods, bacterial deoxyribonucleic acid (DNA) sequences were found using next-generation sequencing in 15/34 (44%) cases. Escherichia coli was present in 6 specimens and was the most frequently identified bacterium. There was a trend towards a higher rate of bacterial DNA positivity in specimens of calcific valves than in those of non-calcific valves (10/17 vs 5/17, P = 0.17). Based on a quantitative test, E. coli accounted for 0.7% ± 1% in calcific valvular tissue and 0.3% ± 0.3% in non-calcific valvular tissue (P = 0.2), and for 11% ± 27% in the valvular tissue of diabetic patients and 0.3% ± 1% in the valvular tissue of non-diabetic patients (P = 0.08). Detection of bacterial DNA in non-endocarditis valvular tissues could be a relatively common finding. There could be an association between the valvular microbiome and certain models of valve degeneration and common metabolic disorders.

Research on sleeve gastrectomy for the treatment of rats with type 2 diabetes mellitus and the regulation of ghrelin and intestinal lesions.

To explore the effect of sleeve gastrectomy on type 2 diabetes mellitus (T2DM) in regulating ghrelin and intestinal lesions. Specific pathogen free (SPF) Wistar rats were injected with streptozotocin (STZ) after giving a high-sugar and high-fat diet, to establish a T2DM rat model. The rats were randomly divided into a sleeve gastric excision group, a non-surgical group and a fake surgical group, with 10 rats in each group. The weight, blood glucose, glucose tolerance and ghrelin hormone of rats were compared. The feces of rats in each group at the 8th week after surgery were collected, to extract the total bacterial deoxyribonucleic acid (DNA). The bacterial 16S universal primer was used to expand the 16SrRNA V46 conserved region. The total Polymerase ChainReaction (PCR) products were sequenced by PE101-bp to classify the gene and genera. The weight of the rats after sleeve gastrectomy significantly decreased (p <0.05). The area under the blood glucose curve and the area under the insulin curve were significantly smaller than those in the non-surgical group and the fake surgical group (p <0.05). Compared with the sleeve gastric excision group, the abundance of PhylumFirmicutes was higher, that of Bacteroidetes was lower. Compared with the sleeve gastric excision group, there were more genera in the fake surgical group and the non-surgical group. The genera with higher abundance in the three groups were Lactobacillus and Bacteroidetes. Compared with the sleeve gastric excision group, the fake surgical group and the non-surgical group had higher abundance of PhylumFirmicutes (p <0.05) and lower abundance of Bacteroidetes (p <0.05). To sum up, sleeve gastrectomy can reduce the weight of rats in T2DM rat model, lower blood glucose levels of rats in the model and improve insulin resistance levels. The related mechanism may be related to the upregulation of ghrelin and intestinal flora.

Disequilibrium of Plasma Protease/Anti-Protease Due to Severe Periodontal Disease Contributes to Human Subarachnoid Hemorrhage

ABSTRACT BACKGROUND The pathophysiology of subarachnoid hemorrhages (SAHs) due to ruptured intracranial aneurysms (IAs) remains unclear. Although a relationship between SAHs and periodontal disease (PD) has been suggested, the mechanism requires clarification. OBJECTIVE To evaluate the relationship between PD and SAHs and to identify periodontal pathogens associated with SAHs. METHODS This prospective study included consecutive patients with ruptured (n = 11) and unruptured (n = 14) IAs and healthy controls (n = 8). The plasma and plaque subgingival bacterial deoxyribonucleic acid (DNA) levels in PD were evaluated by a dentist using the Community Periodontal Index of Treatment Needs (CPITN). Plasma levels of matrix metalloproteinase (MMP-9), tissue inhibitors of matrix metalloproteinase (TIMP2), and procollagen I were analyzed. RESULTS Patients with ruptured IAs, had significantly higher CPITN scores than the controls, suggesting that ruptured IAs were associated with severe PD. Although no rupture-specific bacteria were identified, the positive rate of plaque subgingival bacterial DNA was significantly higher in patients with severe PD than in those without severe PD. Multivariate logistic regression analysis indicated that bleeding on probing (BOP) was associated with ruptured IAs (odds ratio, 1.10; 95% confidence interval 1.04–1.20; P = .0001). BOP was positively associated with plasma MMP-9 levels and a disequilibrium in the MMP-9/TIMP2 ratio. BOP was negatively correlated with plasma procollagen I levels (P &lt; .05, for each). This suggested that local inflammation with severe PD might have systemic effects and lead to ruptured IAs. CONCLUSION Disequilibrium of plasma protease/anti-protease associated with a high BOP rate in severe PD may be attributable to IA rupture.