Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal.

To determine the antibacterial effect of propolis nanoparticles (PNs) as an endodontic irrigant against Enterococcus faecalis biofilm inside the endodontic root canal system. Two-hundred-ten extracted human teeth were sectioned to obtain 6 mm of the middle third of the root. The root canal was enlarged to an internal diameter of 0.9 mm. The specimens were inoculated with E. faecalis for 21 days. Following this, specimens were randomly divided into seven groups, with 30 dentinal blocks in each group including: group I-saline; group II-propolis 100 µg/mL; group III-propolis 300 µg/mL; group IV-propolis nanoparticle 100 µg/mL; group V-propolis nanoparticle 300µg/mL; group VI-6% sodium hypochlorite; group VII-2% chlorhexidine. Dentin shavings were collected at 200 and 400 µm depths, and total numbers of CFUs were determined at the end of one, five, and ten minutes. The non-parametric Kruskal-Wallis and Mann-Whitney tests were used to compare the differences in reduction in CFUs between all groups, and probability values of p < 0.05 were set as the reference for statistically significant results. The antibacterial effect of PNs as an endodontic irrigant was also assessed against E. faecalis isolates from patients with failed root canal treatment. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were also performed after exposure to PNs. A Raman spectroscope, equipped with a Leica microscope and lenses with curve-fitting Raman software, was used for analysis. The molecular interactions between bioactive compounds of propolis (Pinocembrin, Kaempferol, and Quercetin) and the proteins Sortase A and ß-galactosidase were also understood by computational molecular docking studies. PN300 was significantly more effective in reducing CFUs compared to all other groups (p < 0.05) except 6% NaOCl and 2% CHX (p > 0.05) at all time intervals and both depths. At five minutes, 6% NaOCl and 2% CHX were the most effective in reducing CFUs (p < 0.05). However, no significant difference was found between PN300, 6% NaOCl, and 2% CHX at 10 min (p > 0.05). SEM images also showed the maximum reduction in E. faecalis with PN300, 6% NaOCl, and 2% CHX at five and ten minutes. CLSM images showed the number of dead cells in dentin were highest with PN300 compared to PN100 and saline. There was a reduction in the 484 cm-1 band and an increase in the 870 cm-1 band in the PN300 group. The detailed observations of the docking poses of bioactive compounds and their interactions with key residues of the binding site in all the three docking protocols revealed that the interactions were consistent with reasonable docking and IFD docking scores. PN300 was equally as effective as 6% NaOCl and 2% CHX in reducing the E. faecalis biofilms.

Using easy-to-biodegrade co-substrate to eliminate microcystin toxic on electrochemically active bacteria and enhance bioelectricity generation from cyanobacteria biomass.

Cyanobacterial biomass is a promising natural resource for power generation, through the reactions bio-catalyzed by electrochemically active bacteria (EAB). However, the major limitation is the involvement of Microcystin-LR (MC-LR) in inhibiting EAB activation. In this work, toxic M. aeruginosa biomass was employed as analyte of a microbial fuel cell (MFC), and sodium acetate was applied as easy-to-biodegrade co-substrate to alleviate the MC-LR stress on EAB survival. The running stability was continuously enhanced with the increment of co-substrate concentration. The sufficient co-substrate supply (6.0 mM) eliminated the negative effects of MC-LR on the cyanobacteria biomass fed-MFC performance; it contributed 12.7% extension on the electric cyclic terms and caused the productions of the power density which was comparable and even 3.8% higher than its corresponding control (MFC treated with acetate alone). The co-substrate addition also increased coulombic efficiency by 60.1%, microcystin-LR removal efficiency increased by 64.7%, and diversified the microbial community with more species able to biodegrade the MC-LR, bio-transforming the metabolites and EAB. Microcystin-degrading bacteria, such as Sphingopyxis sp., Burkholderia-Paraburkholderia, and Bacillus sp., were remarkably increased, and EAB, including Shewanella sp., Desulfovibrio desulfuricans, Aeromonas hydrophila, were also much more enriched in co-substrate use protocol. Therefore, this study verified a co-substrate strategy for simultaneously eliminating MC-LR toxin and enhancing bioelectricity generation from cyanobacterial biomass via an MFC.

Effectiveness of chitosan-propolis nanoparticle against Enterococcus faecalis biofilms in the root canal.

BACKGROUND: The successful outcome of endodontic treatment depends on controlling the intra-radicular microbial biofilm by effective instrumentation and disinfection using various irrigants and intracanal medicaments. Instrumentation alone cannot effectively debride the root canals specially due to the complex morphology of the root canal system. A number of antibiotics and surfactants are being widely used in the treatment of biofilms however, the current trend is towards identification of natural products in disinfection. The aim of the study was to determine the antibacterial effect of chitosan-propolis nanoparticle (CPN) as an intracanal medicament against Enterococcus faecalis biofilm in root canal. METHODS: 240 extracted human teeth were sectioned to obtain 6 mm of the middle third of the root. The root canal was enlarged to an internal diameter of 0.9 mm. The specimens were inoculated with E. faecalis for 21 days. Following this, specimens were randomly divided into eight groups (n = 30) according to the intracanal medicament placed: group I: saline, group II: chitosan, group III: propolis100 µg/ml (P100), group IV: propolis 250 µg/ml (P250), group V: chitosan-propolis nanoparticle 100 µg/ml (CPN100), group VI: chitosan-propolis nanoparticle 250 µg/ml (CPN250), group VII: calcium hydroxide(CH) and group VIII: 2% chlorhexidine (CHX) gel. Dentine shavings were collected at 200 and 400 µm depths, and total numbers of CFUs were determined at the end of day one, three and seven. The non-parametric Kruskal Wallis and Mann-Whitney tests were used to compare the differences in reduction of CFUs between all groups and probability values of p < 0.05 were set as the reference for statistically significant results. The scanning electron microscope (SEM) and confocal laser scanning microscopy (CLSM) were also performed after exposure to CPNs. The effectiveness of CPNs were also evaluated against E. faecalis isolated obtained from patients having failed root canal treatment. RESULTS: The treatments of chitosan, P100, P250, CPN100, CPN250, CH and 2% CHX reduced the CFUs significantly compared to saline (p < .05). On day one and three, at 200 and 400-µm, CPN250 showed significant reduction of CFUs compared to all other groups (p < .05), while CPN100 was significantly better than other groups (p < .05) except CPN250 and 2% CHX. On day seven, at 200-µm CPN250 showed significant reduction of CFUs compared to all other groups (p < .05) except CPN100 and CHX, while at 400 µm CPN250 showed similar effectiveness as CPN100, CH and 2% CHX. SEM images showed root canal dentin treated with CPN250 had less coverage with E. faecalis bacteria similarly, CLSM images also showed higher percentage of dead E. faecalis bacteria with CPN250 than to CPN100. CONCLUSION: CPN250 was the most effective in reducing E. faecalis colonies on day one, three at both depths and at day seven CPN250 was equally effective as CPN100 and 2% CHX.

Antimicrobial sensitivity pattern, demographic findings and risk factors amongst meningitis and non-meningitis invasive Streptococcus pneumoniae at Aga Khan University Hospital Clinical Laboratory, Karachi, Pakistan.

OBJECTIVE: To determine the resistance rate of penicillin and ceftriaxone amongst invasive meningitis and nonmeningitis isolates of streptococcus pneumoniae. METHODS: The prospective cross-sectional study was conducted from January 2011 to March 2014 at the Clinical Microbiology Laboratory of Aga Khan University, Karachi, and comprised all invasive strains of streptococcus pneumoniae. Penicillin and ceftriaxone susceptibilities were performed and interpreted based on minimum inhibitory concentration breakpoints recommended by Clinical and Laboratory Standards Institute guidelines. Data was analysed using Stata 12. RESULTS: There were 163 strains isolated from sterile body fluids of 109 patients. Of the total, 46(28%) samples were meningitic while 117(72%) were non-meningitic. Of the meningeal isolates, 12(26%) were resistant to penicillin, while none was resistant to ceftriaxone and vancomycin. None of non meningeal isolates showed resistance to penicillin, ceftriaxone or vancomycin. CONCLUSION: There was considerable penicillin resistance among meningeal strains of streptococcus pneumoniae, but here appeared to be no need to add vancomycin for empirical treatment of invasive streptococcus pneumonia infection.

Severity of Caregiver Stress in Relation to Severity of Disease in Persons with Parkinson's.

Introduction Parkinson's disease (PD) is a progressive neurological disorder. It presents with motor symptoms and gradually progresses to cognitive impairment. It has debilitating impact not only on the psychological health of the patient but also of the caregivers. The aim of this study is to evaluate the stress level among caregivers of PD and assess its correlation with the disease factors including duration, severity, and presence of on-off phenomenon. Methods One hundred and fifty-six patients of Parkinson's disease and their caregivers were enrolled. Severity of Parkinson's disease was assessed using Hoehn and Yahr Scale. Caregiver stress was evaluated using Caregiver Burden Inventory. Data was entered and analyzed using Statistical Package for the Social Sciences (SPSS) version 22.0 (IBM Corp., Armonk, NY, USA). Results There were 112 (71.8%) women and 44 (28.2%) men in the caregiver group. Their mean age was 47.75 ± 11.98 years. There were 98 (62.8%) stressed caregivers and 58 (37.2%) non-stressed caregivers. In patients with stage 4 and 5 PD, 67-80% caregivers were stressed as compared to only 28% caregivers of stage 1 PD (p-value = 0.0008). Duration of Parkinson's disease more than 10 years and presence of on-off phenomenon was also significantly associated with higher stress in the caregivers (p-value < 0.00001; p-value = 0.002, respectively). Among the stressed caregivers, 85 (86.7%) were women and only 13 (13.3%) were men (p-value < 0.0001). Conclusion Psychological health of caregivers of persons with Parkinson's is bleak. As the disease progresses, they further succumb to debilitating stress and depression. Qualitative and quantitative studies must be conducted in Pakistan to understand the psychosocial status of Parkinson's disease caregivers and plan strategies to improve their quality of life.

Antimicrobial Susceptibility Patterns of Salmonella Species in Southern Pakistan.

Introduction Typhoid fever is a major infectious disease among the pediatric population of Pakistan. With inappropriate use of antibiotics and rising trends of multidrug-resistant (MDR) and extended drug-resistant (XDR) typhoid, it is becoming a public health emergency. This study evaluated the current trends in antibiotic susceptibilities to Salmonella (S) typhi and paratyphi A, B, and C in southern Pakistan. Materials and methods This cross-sectional study, conducted in the Pediatrics Department, Civil Hospital, Jamshoro from July to December 2018, included children with S. typhi and S. paratyphi A and B strains isolated from the laboratory-based culture of blood samples. Results There were 223 (81.1%) children with S. typhi and 52 (18.9%) with S. paratyphi isolates. Their mean age was 5 ± 3 years. The most common age group with S. typhi strains was two to five years (n = 102; 37.1%). Previous trials of antibiotics were taken by 162 (58.9%) children; 65 (40.1%) of these were physician-prescribed. Cefixime was most commonly taken (66.6%), followed by ciprofloxacin (33.3%). Cefixime and ceftriaxone showed 60.9% and 65.8% sensitivity, respectively. Ciprofloxacin sensitivity was seen in 50.1% S. typhi isolates. There were six (2.6%) cases of MDR typhoid and two (0.9%) cases of XDR typhoid. Conclusion Resistance to second-line antityphoid agents is increasing. Therefore, there is a need to modify prescribing behavior. The outbreak of XDR typhoid among children is an alarming public health concern for Pakistan. Widespread antibiotic stewardship programs must be conducted.

Circadian Variation In The Onset Of Acute Myocardial Infarction In Diabetics.

BACKGROUND: It has been shown in previous studies there is circadian variation in the onset of acute myocardial infarction. The objective of this study was to evaluate the relation of circadian variation in onset of Acute Myocardial infarction in Diabetic subjects. METHODS: This study was conducted at the Services Institute of Medical Sciences Lahore and Punjab Institute of Cardiology from January 2015 to February 2016. Hundred diabetic and 100 Non-diabetic patients with Myocardial infarction were included in the study. Among diabetics those were included in the study that had diabetes for ≥5 years. The time of onset of symptoms to determine the circadian rhythm was noted. In order to determine the frequency of acute myocardial infarction associated with circadian rhythm, 24 hours of the day were divided into four equal sections of 6 hours each. We noted time of onset of acute MI. Thereafter, patients were bracketed in their respective six-hour time periods. These six-hour periods were 0-6, 6:01-12, 12:01-18, and 18:01-24 hours. RESULTS: In this study patients' mean age was 59.16±13.81. Forty-two (71.2%) non-diabetic patients had acute myocardial infarction (AMI) during 6:00-12:00 hours whereas 17 (28.8%) diabetic patients presented with AMI during this time. CONCLUSIONS: Incidence of AMI is significantly increased in the morning 6:01-12:00 hours in non-diabetics. However, diabetic subjects did not show significant increased incidence of AMI during this time rather there was increased incidence of AMI during 0-6:00 hours.

Agreement of Direct Antifungal Susceptibility Testing from Positive Blood Culture Bottles with the Conventional Method for Candida Species.

Early availability of antifungal susceptibilities can ensure timely institution of targeted therapy in candidemia, which can improve patient outcomes. This study prospectively determines the agreement between the results of direct testing of antifungal susceptibilities from blood culture bottles by disk diffusion and Etest and the results of standardized susceptibility testing methods; direct testing would allow susceptibility results to be available 1 to 2 days earlier. A total of 104 blood cultures with different Candida species (28% C. albicans, 27% C. parapsilosis, 26% C. tropicalis, etc.) were evaluated between January 2012 and May 2013 for agreement of fluconazole, voriconazole, and amphotericin B susceptibility results by disk diffusion. Agreement in MICs obtained by Etest was determined for fluconazole (21 isolates), voriconazole (28 isolates), amphotericin (29 isolates), and caspofungin (29 isolates). The kappa scores for categorical agreement were highest for fluconazole by disk diffusion (0.902, standard error [SE] = 0.076) and Etest (1.00, SE = 0.218) and for amphotericin B by disk diffusion (1.00, SE = 0.098). The Pearson correlation (r) of zone diameters was strongest for fluconazole (0.69) and amphotericin (0.70) and moderate for voriconazole (0.60), and the Pearson correlation of MICs was strongest for fluconazole (0.94) and caspofungin (0.88). However, the moderate correlation of amphotericin MICs with zone diameters (-0.42) precludes the use of amphotericin B disk diffusion for susceptibility testing. There were no very major errors; however, there were 1 (1%) major and 5 (4.8%) minor errors with disk diffusion and 4 (13.3%) minor errors with Etest. Thus, antifungal disk diffusion directly from blood culture bottles is a rapid and easy method for fluconazole and voriconazole susceptibility testing for timely tailoring of candidemia therapy.

Characteristics and outcomes in community-acquired versus hospital-acquired acute kidney injury.

AIM: Relatively little is known about the prevalence of acute kidney injury developing outside a hospital setting (CA-AKI) or the impact of CA-AKI on short-term or long-term clinical outcomes. The objective of this study was to compare the prevalence, causes, severity and outcomes of patients with CA-AKI and hospital-acquired (HA)-AKI. METHODS: A retrospective cohort study of patients with AKI identified by ICD-9 code at a single VA (Veterans Affairs) hospital from September 1999 to May 2007 was performed. AKI was verified by applying the RIFLE criteria, and patients were categorized as CA-AKI if RIFLE criteria were met at admission. Demographic, clinical, and outcome variables were extracted by chart review. RESULTS: Four hundred twenty-two patients met inclusion criteria, of which 335 (79.4%) developed CA-AKI. Patients with CA-AKI were more likely to have volume depletion as the aetiology, had fewer chronic illnesses and hospital complications, had a shorter length of stay, and had a reduced mortality, compared with HA-AKI. Distribution among the three RIFLE classes did not differ between groups, and recovery of renal function was incomplete in both groups. CONCLUSION: We conclude that CA-AKI is a common cause of AKI that is as severe as that seen in HA-AKI. CA-AKI has a significant impact on length of hospital stay, mortality, and the development and/or progression of chronic kidney disease. Strategies to limit the risk of CA-AKI are likely to have a significant impact on healthcare costs and patient care.

Effect of L-arginine therapy on plasma NO2 and NO3 levels, and blood pressure in uremic rabbits.

BACKGROUND: Normal kidney function is regulated by Nitric oxide (NO) and Superoxide (O2-) in the body, and consequently controls blood pressure. Nitric Oxide promotes natriuresis and diuresis, and therefore results in reduction of blood pressure. The objective of this study was to determine the effect of L-arginine supplementation on blood pressure, urinary protein, nitrite and nitrate in addition to blood urea, serum creatinine and creatinine clearance in uremic rabbits. METHODS: This study was carried out in the Department of Biochemistry Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre, Karachi. A total of 48 rabbits were included in the study. Twenty-four of the rabbits on surgical intervention were prepared as uremic and so became hypertensive as well. Two groups were uremic, one group was given L-arginine and the other was remained untreated. Systolic and diastolic blood pressure was measured on week 0, week 2, week 4, and week 6, while blood and urine was collected on week 0 and week 6. RESULTS: On supplementation with L-arginine to uremic rabbits systolic and diastolic blood pressures were decreased significantly. Nitrite/nitrate and urinary protein were corrected to some extent while blood urea and serum creatinine were unaffected. CONCLUSION: L-arginine has a beneficial role as blood pressure lowering agent in uremic rabbits. It corrects NO2/NO3 plasma level and proteinuria which is indicator of renal failure.

Survival of patients treated with intra-aortic balloon counterpulsation at a tertiary care center in Pakistan - patient characteristics and predictors of in-hospital mortality.

BACKGROUND: Intra-aortic balloon counterpulsation (IABC) has an established role in the treatment of patients presenting with critical cardiac illnesses, including cardiogenic shock, refractory ischemia and for prophylaxis and treatment of complications of percutaneous coronary interventions (PCI). Patients requiring IABC represent a high-risk subset with an expected high mortality. There are virtually no data on usage patterns as well as outcomes of patients in the Indo-Pakistan subcontinent who require IABC. This is the first report on a sizeable experience with IABC from Pakistan. METHODS: Hospital charts of 95 patients (mean age 58.8 (+/- 10.4) years; 78.9% male) undergoing IABC between 2000-2002 were reviewed. Logistic regression was used to determine univariate and multivariate predictors of in-hospital mortality. RESULTS: The most frequent indications for IABC were cardiogenic shock (48.4%) and refractory ischemia (24.2%). Revascularization (surgical or PCI) was performed in 74 patients (77.9%). The overall in-hospital mortality rate was 34.7%. Univariate predictors of in-hospital mortality included (odds ratio [95% CI]) age (OR 1.06 [1.01-1.11] for every year increase in age); diabetes (OR 3.68 [1.51-8.92]) and cardiogenic shock at presentation (OR 4.85 [1.92-12.2]). Furthermore, prior CABG (OR 0.12 [0.04-0.34]), and in-hospital revascularization (OR 0.05 [0.01-0.189]) was protective against mortality. In the multivariate analysis, independent predictors of in-hospital mortality were age (OR 1.13 [1.05-1.22] for every year increase in age); diabetes (OR 6.35 [1.61-24.97]) and cardiogenic shock at presentation (OR 10.0 [2.33-42.95]). Again, revascularization during hospitalization (OR 0.02 [0.003-0.12]) conferred a protective effect. The overall complication rate was low (8.5%). CONCLUSIONS: Patients requiring IABC represent a high-risk group with substantial in-hospital mortality. Despite this high mortality, over two-thirds of patients do leave the hospital alive, suggesting that IABC is a feasible therapeutic device, even in a developing country.