Does Platelet-Rich Plasma Deserve a Role in Accelerating the Recovery of Reflex Sympathetic Dystrophy Following Distal Radius Fracture?

Introduction: This study was to evaluate the efficacy of multiple platelet-rich plasma injections in reflex sympathetic dystrophy following distal radius fracture after previous various treatments have failed. Materials and methods: This comparative prospective study was designed for 64 patients of reflex sympathetic dystrophy developed following distal radius fracture, from January 2009 to December 2020 were enrolled in this study. This cohort of patient was given either four multiple subcutaneous platelet-rich plasma injections at weekly interval (n = 32) or two injections in a month with 15 days interval (n = 32). The primary outcome measure assessed with patient rated wrist evaluation questionnaire score. The secondary outcome was a visual analogue scale pain score. The final follow up was at 2 years. p ≤ 0.05 is considered statistically. Results: The patient rated wrist evaluation score for usual and specific activities and EQ-VAS for pain level showed statistically significant greater improvement in group A (42 ± 21%) compared to group B (19 ± 24%), (p = 0.37). Patients also had improvement in wrist movements with no statistically significant differences in both groups. The standard difference in means of all three functional scores was almost similar between both groups A and B (standard difference in means = 0.032; 95% CI 0.236-0.830; p = 0.495), considered clinically meaningful. Conclusion: This study results suggest autologous platelet-rich plasma injections seem to be safe, cost effective, efficacious algorithm treatment for reflex sympathetic dystrophy following distal radius fracture patients where previous treatments have failed.

Optimisation of enzyme-assisted extraction of camptothecin from Nothapodytes nimmoniana and its characterisation.

INTRODUCTION: Efficient extraction of camptothecin (CPT), an anticancer agent from the commercial source Nothapodytes nimmoniana (J. Graham) Mabb in India, is of paramount importance. CPT is present in the highest concentration in the stem portion, and the stem can be readily harvested without uprooting the plant. The fluorescence microscopy mapping of the bark matrix for CPT revealed its presence in a free form within both the outer (epidermal and cortical tissues) and inner (xylem and phloem tissues) sections. The bark matrix primarily consists of cellulose, hemicellulose, and lignin, rendering it woody, rigid, and resistant to efficient solvent penetration for CPT extraction. We proposed a hypothesis that subjecting it to disruption through treatment with hydrolytic enzymes like cellulase and xylanase could enhance solvent diffusion, thereby enabling a swift and effective extraction of CPT. OBJECTIVE: The present study was aimed at enzyme-assisted extraction, using cellulase and xylanase for hydrolytic disruption of the cells to readily access CPT from the stem of the plant N. nimmoniana (J. Graham) Mabb. METHODOLOGY: The hydrolytic cell disruption of ground powder from the stem bark was studied using cellulase and xylanase enzymes. The enzymatically pretreated stem bark powder was subsequently recovered by filtration, dried, and subjected to extraction with methanol to isolate CPT. This process was optimised through a Box-Behnken design, employing a one-factor-at-a-time approach to assess parameters such as enzyme concentration (2-10% w/w), pH (3-7), incubation time (6-24 h), and solid-to-solvent ratio (1:30-1:70 g/mL). CPT was characterised using proton nuclear magnetic resonance (1H-NMR) and Fourier transform infrared (FTIR) spectra, and a high-performance liquid chromatography (HPLC) method was developed for quantification. RESULTS: The cellulase and xylanase treatment resulted in the highest yields of 0.285% w/w and 0.343% w/w, with efficiencies of 67% and 81%, respectively, achieved in a significantly shorter time compared to the untreated material, which yielded 0.18% with an efficiency of only 42%. Extraction by utilising the predicted optimised process parameters, a nearly two-fold increase in the yield, was observed for xylanase, with incubation and solvent extraction times set at 16 and 2 h, respectively. Scanning electron microscopy (SEM) images of the spent material indicated perforations attributed to enzymatic action, suggesting that this could be a primary factor contributing to the enhanced extraction. CONCLUSION: Enzyme-mediated hydrolytic cell disruption could be a potential approach for efficient and rapid isolation of CPT from the bark of N. nimmoniana.

Artificial intelligence: Machine learning approach for screening large database and drug discovery.

Recent research in drug discovery dealing with many faces difficulties, including development of new drugs during disease outbreak and drug resistance due to rapidly accumulating mutations. Virtual screening is the most widely used method in computer aided drug discovery. It has a prominent ability in screening drug targets from large molecular databases. Recently, a number of web servers have developed for quickly screening publicly accessible chemical databases. In a nutshell, deep learning algorithms and artificial neural networks have modernised the field. Several drug discovery processes have used machine learning and deep learning algorithms, including peptide synthesis, structure-based virtual screening, ligand-based virtual screening, toxicity prediction, drug monitoring and release, pharmacophore modelling, quantitative structure-activity relationship, drug repositioning, polypharmacology, and physiochemical activity. Although there are presently a wide variety of data-driven AI/ML tools available, the majority of these tools have, up to this point, been developed in the context of non-communicable diseases like cancer, and a number of obstacles have prevented the translation of these tools to the discovery of treatments against infectious diseases. In this review various aspects of AI and ML in virtual screening of large databases were discussed. Here, with an emphasis on antivirals as well as other disease, offers a perspective on the advantages, drawbacks, and hazards of AI/ML techniques in the search for innovative treatments.

Convolutional neural networks in medical image understanding: a survey.

Imaging techniques are used to capture anomalies of the human body. The captured images must be understood for diagnosis, prognosis and treatment planning of the anomalies. Medical image understanding is generally performed by skilled medical professionals. However, the scarce availability of human experts and the fatigue and rough estimate procedures involved with them limit the effectiveness of image understanding performed by skilled medical professionals. Convolutional neural networks (CNNs) are effective tools for image understanding. They have outperformed human experts in many image understanding tasks. This article aims to provide a comprehensive survey of applications of CNNs in medical image understanding. The underlying objective is to motivate medical image understanding researchers to extensively apply CNNs in their research and diagnosis. A brief introduction to CNNs has been presented. A discussion on CNN and its various award-winning frameworks have been presented. The major medical image understanding tasks, namely image classification, segmentation, localization and detection have been introduced. Applications of CNN in medical image understanding of the ailments of brain, breast, lung and other organs have been surveyed critically and comprehensively. A critical discussion on some of the challenges is also presented.

Comparative evaluation of antimicrobial and antifungal efficacy of bioactive root-end filling materials: An in vitro study.

CONTEXT: Microorganisms are the main cause of failure of endodontic treatment. When retreatment fails periapical surgery followed by retrograde filling is done to seal the apex. A root-end filling material should have antimicrobial property as well as bioactive properties necessary for healing, repair, and regeneration of the apex. AIMS: The aim of this study was to evaluate the antibacterial and antifungal efficacy of three bioactive root-end filling materials: mineral trioxide aggregate (MTA) Plus, Biodentine, Endosequence root repair material (ERRM) against Enterococcus faecalis and Candida albicans. SUBJECTS AND METHODS: E. faecalis and C. albicans standard bacterial strains were used. 100 µl was taken from liquid cultures of E. faecalis and planted in Mueller-Hinton agar and the same amount of C. albicans was planted in Sabouraud dextrose agar by lawn culture. MTA Plus, Biodentine, and ERRM were aseptically filled into the opened pits. Following this, the media were kept in the drying oven at 37°C for 24, 48, and 72 h and the diameters of the inhibition zones were measured. STATISTICAL ANALYSIS USED: Statistical analysis was carried out by Kruskal-Wallis, Post hoc (Mann-Whitney), Friedman, and Post hoc (Wilcoxon-sign) test. RESULTS: Among the three groups, the antimicrobial activity of Biodentine against E. faecalis was statistically higher than MTA Plus and ERRM (P < 0.05). Antifungal activity of MTA Plus against C. albicans was statistically higher than Biodentine and ERRM (P < 0.05). ERRM showed the smallest inhibition zone against E. faecalis and C. albicans among the three groups (P < 0.05). CONCLUSIONS: Biodentine exhibited the greatest antimicrobial activity and MTA Plus exhibited the greatest antifungal activity among the three groups. ERRM exhibited the least antibacterial and antifungal activity among the three groups.

Comparative Assessment of the Antimicrobial Efficacy of Triclosan, Amoxicillin and Eugenol against Enterococcus faecalis.

AIMS: Elimination of microorganisms and prevention of recurrence of infection from the complex root canal system of primary teeth requires an obturating material with broad antimicrobial activity. Hence, the purpose of the study is to assess and compare the antimicrobial efficacy of Triclosan, Amoxicillin and Eugenol individually and in combinations against a resistant microorganism viz., Enterococcus faecalis. MATERIALS AND METHODS: A two-fold serial dilution method was used to check the minimum inhibitory concentration (MIC) of triclosan, amoxicillin and eugenol against thirty E. faecalis (isolated from oral lesions). The resistant strains were subjected to different combinations of three agents by modified checkerboard method. MIC was determined after incubation for 24 hours at 370°C. Then the three dilutions from MIC were inoculated on BHI agar plates and incubated overnight to determine minimum bactericidal concentration (MBC). RESULTS: The mean MIC and MBC of triclosan was 3.43 µg/mL and 3.75 µg/mL respectively. Whereas for amoxicillin, it was 3.43 µg/mL and 3.85 µg/mL. Eugenol did not show any inhibition up to a concentration of 3200 µg/mL. In combination, eugenol showed good synergistic effect with both triclosan and amoxicillin. In combination with triclosan, eugenol showed much promising result as compared with amoxicillin. But triclosan and amoxicillin combination showed inhibition at higher concentrations. CONCLUSION: Triclosan and eugenol combination showed better effectiveness against E. faecalis in comparison to amoxicillin and eugenol. Triclosan and amoxicillin showed antagonism when used in combination against E. faecalis. HOW TO CITE THIS ARTICLE: Gowda J, Tavarageri A, Kulkarni R, et al. Comparative Assessment of the Antimicrobial Efficacy of Triclosan, Amoxicillin and Eugenol against Enterococcus faecalis. Int J Clin Pediatr Dent 2021;14(1):59-62.

Experimentally Validated QSAR Model for Surface pK a Prediction of Heterolipids Having Potential as Delivery Materials for Nucleic Acid Therapeutics.

The application of lipid-based drug delivery technologies for bioavailability enhancement of drugs has led to many successful products in the market for clinical use. Recent studies on amine-containing heterolipid-based synthetic vectors for delivery of siRNA have witnessed the United States Food and Drug Administration (USFDA) approval of the first siRNA drug in the year 2018. The studies on various synthetic lipids investigated for delivery of such nucleic acid therapeutics have revealed that the surface pK a of the constructed nanoparticles plays an important role. The nanoparticles showing pK a values within the range of 6-7 have performed very well. The development of high-performing lipid vectors with structural diversity and falling within the desired surface pK a is by no means trivial and requires tedious trial and error efforts; therefore, a practical solution is called for. Herein, an attempt to is made provide a solution by predicting the statistically significant pK a through a predictive quantitative structure-activity relationship (QSAR) model. The QSAR model has been constructed using a series of 56 amine-containing heterolipids having measured pK a values as a data set and employing a partial least-squares regression coupled with stepwise (SW-PLSR) forward algorithm technique. The model was tested using statistical parameters such as r 2, q 2, and pred_r 2, and the model equation explains 97.2% (r 2 = 0.972) of the total variance in the training set and it has an internal (q 2) and an external (pred_r 2) predictive ability of ∼83 and ∼63%, respectively. The model was validated by synthesizing a series of designed heterolipids and comparing measured surface pK a values of their nanoparticle assembly using a 2-(p-toluidino)-6-napthalenesulfonic acid (TNS) assay. Predicted and measured surface pK a values of the synthesized heterolipids were in good agreement with a correlation coefficient of 93.3%, demonstrating the effectiveness of this QSAR model. Therefore, we foresee that our developed model would be useful as a tool to cut short tedious trial and error processes in designing new amine-containing heterolipid vectors for delivery of nucleic acid therapeutics, especially siRNA.

Low oxygen microenvironment and cardiovascular remodeling: Role of dual L/N.type Ca2+ channel blocker.

OBJECTIVE: Patients exposed to chronic sustained hypoxia frequently develop cardiovascular disease risk factors to ultimately succumb to adverse cardiovascular events. In this context, the present study intends to assess the role of cilnidipine (Cil), a unique calcium channel blocker that blocks both L-type and N-type calcium channels, on cardiovascular pathophysiology in face of chronic sustained hypoxia exposure. MATERIALS AND METHODS: The study involved Wistar strain albino rats. The group-wise allocation of the experimental animals is as follows - Group 1, control (21% O2); Group 2, chronic hypoxia (CH) (10% O2, 90% N); Group 3, Cil + 21% O2; and Group 4, CH (10% O2, 90% N) + Cil (CH + Cil). Cardiovascular hemodynamics, heart rate variability, and endothelial functions (serum nitric oxide [NO], serum endothelial nitric oxide synthase [NOS3], and serum vascular endothelial growth factor [VEGF]) were assessed. Cardiovascular remodeling was studied by histopathological examination of the ventricular tissues, coronary artery (intramyocardial), and elastic and muscular arteries. Normalized wall index of the coronary artery was also calculated. RESULTS AND CONCLUSION: The results demonstrated altered cardiovascular hemodynamics, disturbed cardiovascular autonomic balance, increased levels of VEGF and NOS3, and decreased bioavailability of NO on exposure to chronic sustained hypoxia. The histopathological examination further pointed toward cardiovascular remodeling. Treatment with Cil ameliorated the cardiovascular remodeling and endothelial dysfunction induced by CH exposure, which may be due to its blocking actions on L/N-type of calcium channels, indicating the possible therapeutic role of Cil against CH-induced cardiovascular pathophysiology.

Estimation and correlation of serum and salivary glucose and immunoglobulin A levels and salivary candidal carriage in diabetic and non-diabetic patients.

Background. A correlation has been noted between diabetes mellitus (DM) and changes in the oral cavity. The present study aimed to estimate, compare, and correlate serum and salivary glucose and IgA levels and salivary candidal carriage in diabetic and non-diabetic individuals. Methods. Eighty-eight subjects were categorized into three groups: group 1 (controlled DM; n=27), group 2 (uncontrolled DM; n=32) and group 3 (non-diabetics; n=29). Serum and salivary glucose levels were estimated by glucose oxidase/peroxidase method, serum and salivary IgA by a diagnostic kit, and candidal colonization by inoculating samples into Sabouraud dextrose agar plate. Statistical analyses were carried out by one-way ANOVA, post hoc Tukey tests, and Pearson's correlation coefficient. Results. Significant elevation of serum IgA levels was observed in group 2 compared to group 3 and significant decreases in salivary IgA levels in groups 1 and 2. The candidal carriage was significantly higher in group 2 compared to group 3. Serum glucose and salivary IgA levels showed a significant correlation in group 1. There was a positive correlation between serum/ salivary glucose and serum/salivary IgA levels in group 2. In addition, there was a significant correlation between serum glucose and serum IgA levels in group 3. Conclusion. Saliva could be a potential, non-invasive diagnostic tool to estimate glucose levels. The evaluation of salivary components, like IgA, might be useful in diagnosing and managing oral manifestations in diabetic individuals. Elevated salivary glucose levels contribute to elevated candidal carriage, making individuals susceptible to oral candidiasis.

Effect of L-Ascorbic Acid on Nickel-Induced Alteration of Cardiovascular Pathophysiology in Wistar Rats.

Nickel, a widely used heavy metal is suspected as a cardiotoxic element. The aim of the present study was to assess the possible protective role of l-ascorbic acid on nickel-induced alterations of cardiovascular pathophysiology in male albino rats. Twenty-four albino rats (b.wt. 170-250 g) were randomized into four groups: control; l-ascorbic acid (50 mg/100 g b.wt., orally); NiSO4 (2.0 mg/100 g b.wt., i.p.); NiSO4 with l-ascorbic acid. Cardiovascular electrophysiology, serum and cardiac tissue malondialdehyde (MDA), nitric oxide (NO), ascorbic acid, serum α-tocopherol and serum vascular endothelial growth factor (VEGF) were evaluated. Histopathology of cardiac and aortic tissues was also assessed. NiSO4-treated rats showed a significant increase in heart rate, LF/HF ratio and blood pressure (SBP, DBP and MAP). A significant increase of serum MDA, NO and VEGF in NiSO4 treatment with a concomitant decrease of serum ascorbic acid and α-tocopherol as compared to their respective controls were also observed. Simultaneous supplementation of l-ascorbic acid with NiSO4 significantly decreased LF/HF ratio, BP and oxidative stress parameters, whereas ascorbic acid and α-tocopherol concentration was found to be increased. Histopathology of cardiac and aortic tissues showed nickel-induced focal myocardial hypertrophy and degeneration in cardiac tissue with focal aneurism in aortic tissues. Supplementation with l-ascorbic showed a protective action in both cardiac and aortic tissues. Results indicated the possible beneficial effect of l-ascorbic acid on nickel-induced alteration of the cardiovascular pathophysiology in experimental rats.

A novel ready-to-use dry-reagent polymerase chain reaction for detection of Escherichia coli & Shigella species.

Background & objectives: Polymerase chain reaction (PCR) has wide acceptance for rapid identification of pathogens and also for diagnosis of infectious conditions. However, because of economic and expertise constraints, a majority of small or peripheral laboratories do not use PCR. The objective of the present study was to develop a dry-reagent PCR assay as an alternative to conventional PCR to assess its applicability in routine laboratory practice using malB gene for identification of Escherichia coli as a model. Methods: A total of 184 isolates were selected for the study comprising clinical isolates of E. coli and non-E. coli including Shigella sp. and a few other control strains. The DNA was isolated from all the isolates. The isolated DNA as well as the overnight grown bacterial cultures were subjected to both conventional wet PCR and dry-reagent PCR. Results: The genomic DNA isolated from E. coli showed amplification of malB gene in both conventional wet and dry-reagent PCR and the band was observed at 491 bp. In dry-reagent PCR, the overnight grown E. coli cells also showed positive result. The non-E. coli strains other than Shigella sp. showed negative in both conventional wet and dry-reagent PCR. Shigella sp. showed positive in both conventional wet and dry-reagent PCR. Interpretation & conclusions: Considering the elimination of genomic DNA isolation step, and similar results with the conventional wet PCR, dry-reagent PCR may be a good alternative for the conventional wet PCR.

Membranes for dehydration of alcohols via pervaporation.

Alcohols are the essential chemicals used in a variety of pharmaceutical and chemical industries. The extreme purity of alcohols in many of such industrial applications is essential. Though distillation is one of the methods used conventionally to purify alcohols, the method consumes more energy and requires carcinogenic entertainers, making the process environmentally toxic. Alternatively, efforts have been made to focus research efforts on alcohol dehydration by the pervaporation (PV) separation technique using polymeric membranes. The present review is focused on alcohol dehydration using PV separation technique, which is the most efficient and benign method of purifying alcohols that are required in fine chemicals synthesis and developing pharmaceutical formulations. This review will discuss about the latest developments in the area of PV technique used in alcohol dehydration using a variety of novel membranes.

Novel pH-sensitive interpenetrated network polyspheres of polyacrylamide-g-locust bean gum and sodium alginate for intestinal targeting of ketoprofen: In vitro and in vivo evaluation.

In this report, novel pH-sensitive interpenetrated network (IPN) polyspheres were developed utilizing polyacrylamide-g-locust bean gum (PAAm-g-LBG) in combination with sodium alginate (SA) to achieve intestinal targeted delivery of ketoprofen. PAAm-g-LBG was synthesized under microwave irradiation wherein ceric ammonium nitrate was used as reaction initiator and then conversion of PAAm-g-LBG as pH-sensitive copolymer was carried out by alkaline hydrolysis. The PAAm-g-LBG copolymer was characterized through 1H-NMR, FTIR and elemental analysis. The IPN polyspheres exhibited pH-depended swelling or de-swelling with the alteration of surrounding pH. The in-vitro release of drug from IPN polyspheres was found to be higher (≈ 90%) in phosphate buffer of pH 7.4 in comparison with that in pH 1.2 buffer (10.6%). The in-vivo pharmacokinetic, anti-inflammatory screening and stomach histopathology studies performed on Wistar rats revealed pH sensitivity of IPN polyspheres where ketoprofen was successfully targeted to small intestine resulting in reduced side effects of ketoprofen like ulcer formation, erosion of gastric mucosa and hemorrhages.

Reactive mechanism and the applications of bioactive prebiotics for human health: Review.

Prebiotics plays an important role in improving the growth of gut bacteria and it majorly found in various natural food sources such as fruits and vegetables. Nowadays, the prebiotic sources are added as a supplement in various food products such as dairy products, beverages, health drinks, infant formulae, and meat products. The presence of prebiotics provides various health benefits such as improveing calcium and magnesium absorption, increases bone density, reduces cancer risk, decreases cardiovascular diseases and also improves the immune system.

Novel biocompatible poly(acrylamide)-grafted-dextran hydrogels: Synthesis, characterization and biomedical applications.

An electro-responsive PAAm-g-Dxt copolymer was synthesized and characterized by 1HNMR & FTIR spectroscopy, neutralization equivalent, elemental and thermogravimetric analysis to ascertain the grafting reaction. Further, we developed an electro-responsive transdermal drug delivery system (ETDS) utilizing PAAm-g-Dxt copolymer for rivastigmine tartarate delivery through skin. The ETDS were developed using drug-loaded PAAm-g-Dxt hydrogel as the reservoir, and cross-linked dextran-poly(vinyl alcohol) blend films as rate controlling membranes (RCM). In the absence of electrical stimuli, a small amount of drug was permeated from the ETDS, while in the presence of electrical stimuli, the drug permeability was increased. On application of electric stimulus, the flux was increased by 1.6 fold; drug permeability was enhanced when the strength of applied electric current was raised to 8 mA from 2 mA. The drug permeability characteristics studied under "on-off" stimuli suggested that there was faster drug permeation when electrical stimuli was 'on' and it decreased when electrical stimuli was 'off.' The histopathology study confirmed the altered skin structural integrity after application of electrical stimuli. Hence, the PAAm-g-Dxt based ETDS are useful for transdermal drug delivery triggered by an electric stimulus to deliver on-demand release of drug into systemic circulation.

Integration of biological pre-treatment methods for increased energy recovery from paper and pulp biosludge.

The paper and pulp industry (PPI) produces high quantities of solid and liquid discharge and is regarded as the most polluting industry in the world causing adverse effects to environments and human beings. Hence changes in the way PPI sludge and waste materials are treated is urgently required. Nearly, 10 million tons of waste is generated per year, however PPI waste is enriched with many organic chemicalscontaining a high percentage of lignin, cellulose, and hemicellulose which can be used as valuable raw materials for the production of bioenergy and value-added chemicals. Pretreatment of complex lignocellulosic materials of PPI waste is difficult because of the cellulose crystallinity and lignin barrier. At present most of this waste is recycled in a conventional treatment approach through biological and chemical processes, incurring high cost and low returns. Henceefficient pretreatment techniques are required by which complete conversion of PPI waste is possible. Therefore, the present chapter provides the scope of integration of pretreatment methods through which bioenergy recovery is possible during the PPI waste treatment. Detailed information is presented on the various pre-treatment techniques (chemical, mechanical, enzymatic and biological) in order to increase the efficiency of PPI waste treatment and energy recovery from PPI waste. Along with acid and alkali based efficient chemical treatment process, physical methods (i.e. shearing, high-pressure homogenization, etc.), biochemical techniques (whole cell-based and enzyme-based) and finally biological techniques (e.g. aerobic and anaerobic treatment) are discussed. During each of the treatment processes, scope of energy recovery and bottlenecks of the processes were elaborated. The review thus provides systemic insight into developing efficient pretreatment processes which could increase carbon recovery and treatment efficiency of PPI waste.

Identification of multiple strains of Porphyromonas gingivalis using heteroduplex polymerase chain reaction in varying severity of chronic periodontitis.

AIM: Research has demonstrated that there are multiple strains of Porphyromonas gingivalis with varying potency to cause periodontal disease. The current study aims at using heteroduplex polymerase chain reaction (PCR) to detect the strain diversity of P. gingivalis in periodontitis lesions of varying severity in a sample of the Indian population. MATERIALS AND METHODS: Subgingival plaque samples were collected from 60 individuals with varying severity of chronic periodontitis and 30 individuals with a clinically healthy periodontium. The samples were subjected to PCR analysis to identify P. gingivalis, followed by heteroduplex analysis to identify the strain diversity in a given sample. Bacterial culture was carried out as a comparative standard. RESULTS: Of the 56 samples that were positive for P. gingivalis by PCR, 54 samples yielded eight different heteroduplex patterns. Analysis of these patterns indicated that two strains of P. gingivalis were present in 41 individuals (45.6%) and three strains were present in 13 individuals (14.4%). Detection of P. gingivalis by PCR was significantly more in the periodontitis group as compared to the healthy group. CONCLUSIONS: Species-specific PCR and heteroduplex analysis provide a simple and accurate method to analyse the strain diversity of P. gingivalis. P. gingivalis was detected in both healthy periodontal sites as well as sites with periodontitis. The presence of two or three P. gingivalis strains was seen in 60% of the samples.

Bacterial Biofilms in Chronic Rhinosinusitis and Their Implications for Clinical Management.

To study the microbiological profile in patients with chronic rhino-sinusitis. To correlate disease severity with the presence of biofilms and host risk factors. To assess outcome of Sinus Surgery 2 weeks post operatively in terms of presence of bacteria and their ability to form biofilm. Prospective study. 50 cases of chronic rhino-sinusitis requiring Functional Endoscopic Sinus Surgery admitted in SDM Hospital, Dharwad, Karnataka were studied using intra-operative mucosal samples for microbiological analysis. The organisms isolated were tested for biofilm forming ability using three in vitro tests. Severity of disease was assessed using SNOT 22 scoring system. Of 50 cases studied, 66% showed presence of chronic rhino-sinusitis with polyposis and had higher SNOT scores compared to those without polyps. Bacterial isolates were obtained from only 17 samples. Staphylococcus species was isolated from 16 samples and Klebsiella pneumoniae from one. 11 Staph spp. isolates showed biofilm forming ability in vitro. Postoperative events in 3 cases yielded biofilm-forming Staphylococcus. Staphylococcus was the most dominant organism isolated and 11 isolates were biofilm formers. Thus the detection of biofilm forming organisms can be considered as a negative prognostic indicator and should forewarn the surgeon about the risk of recurrence.

Development of a dry-reagent mix-based polymerase chain reaction as a novel tool for the identification of Acinetobacter species and its comparison with conventional polymerase chain reaction.

BACKGROUND: Nosocomial infections are often caused by multidrug-resistant bacteria and the incidence is increasing. Acinetobacter, a Gram-negative bacillus, is commonly associated with the use of intravascular catheterization and airway intubation. Polymerase chain reaction (PCR) for identification of Acinetobacter baumannii from samples has been standardized that use conventional wet-reagent mix. We have designed and optimized a dry-reagent mix for identification of Acinetobacter species by PCR. The dry-reagent mix can be stored at room temperature, has less chances of contamination, and thus can be used at point-of-care diagnosis. AIM AND OBJECTIVE: The present work was focused on comparing the sensitivity and specificity of dry-reagent PCR mix over conventional wet-reagent PCR mix for identification of Acinetobacter species. MATERIALS AND METHODS: Conventional wet-reagent mix based and dry-reagent mix based PCR were carried out for the DNA isolated from Acinetobacter species. The latter was also applied directly on bacterial growth without prior DNA extraction process. Equal numbers of bacterial isolates other than Acinetobacter species were also subjected to identification by the same protocols for determining the sensitivity and specificity of the test. RESULTS: The Acinetobacter species showed amplification of the target rpoB gene and the band was observed at 397 bp. The dry-reagent PCR mix results matched completely with the conventional wet-reagent PCR mix assay. All the non-Acinetobacter isolates were negative for the PCR. This indicates that the test is highly specific. The dry-reagent mix also contained an enzyme resistant to PCR inhibitors and capable of amplifying DNA directly from cells. CONCLUSION: Performance of dry-reagent PCR mix without the need for DNA extraction and preparation of a PCR mix proved to be more sensitive and reduce the handling error, minimizes the time, manual work, and skilled labor.