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In Vitro Bactericidal and Virucidal Efficacy of Povidone-Iodine Gargle/Mouthwash Against Respiratory and Oral Tract Pathogens.

IntroductionRecent virus epidemics and rising antibiotic resistance highlight the importance of hygiene measures to prevent and control outbreaks. We investigated the in vitro bactericidal and virucidal efficacy of povidone-iodine (PVP-I) 7% gargle/mouthwash at defined dilution against oral and respiratory tract pathogens.MethodsPVP-I was tested against Klebsiella pneumoniae and Streptococcus pneumoniae according to bactericidal quantitative suspension test EN13727 and against severe acute respiratory syndrome and Middle East respiratory syndrome coronaviruses (SARS-CoV and MERS-CoV), rotavirus strain Wa and influenza virus A subtype H1N1 according to virucidal quantitative suspension test EN14476. PVP-I 7% gargle/mouthwash was diluted 1:30 with water to a concentration of 0.23% (the recommended concentration for “real-life” use in Japan) and tested at room temperature under clean conditions [0.3 g/l bovine serum albumin (BSA), viruses only] and dirty conditions (3.0 g/l BSA + 3.0 ml/l erythrocytes) as an interfering substance for defined contact times (minimum 15 s). Rotavirus was tested without protein load. A ≥ 5 log10 (99.999%) decrease of bacteria and ≥ 4 log10 (99.99%) reduction in viral titre represented effective bactericidal and virucidal activity, respectively, per European standards.ResultsPVP-I gargle/mouthwash diluted 1:30 (equivalent to a concentration of 0.23% PVP-I) showed effective bactericidal activity against Klebsiella pneumoniae and Streptococcus pneumoniae and rapidly inactivated SARS-CoV, MERS-CoV, influenza virus A (H1N1) and rotavirus after 15 s of exposure.ConclusionPVP-I 7% gargle/mouthwash showed rapid bactericidal activity and virucidal efficacy in vitro at a concentration of 0.23% PVP-I and may provide a protective oropharyngeal hygiene measure for individuals at high risk of exposure to oral and respiratory pathogens.FundingMundipharma Research GmbH & Co. KG (MRG).

Increased intracellular activity of MP1102 and NZ2114 against Staphylococcus aureus in vitro and in vivo

Treatment of Staphylococcus aureus infections remains very difficult due to its capacity to survive intracellularly and its multidrug resistance. In this study, the extracellular/intracellular activities of plectasin derivatives-MP1102/NZ2114 were investigated against three methicillin-susceptible/-resistant S. aureus (MSSA/MRSA) strains in RAW 264.7 macrophages and mice to overcome poor intracellular activity. Antibacterial activities decreased 4–16-fold under a mimic phagolysosomal environment. MP1102/NZ2114 were internalized into the cells via clathrin-mediated endocytosis and macropinocytosis and distributed in the cytoplasm; they regulated tumor necrosis factor-α, interleukin-1β and interleukin-10 levels. The extracellular maximal relative efficacy (Emax) values of MP1102/NZ2114 towards the three S. aureus strains were >5-log decrease in colony forming units (CFU). In the methicillin-resistant and virulent strains, MP1102/NZ2114 exhibited intracellular bacteriostatic efficacy with an Emax of 0.42–1.07-log CFU reduction. In the MSSA ATCC25923 mouse peritonitis model, 5 mg/kg MP1102/NZ2114 significantly reduced the bacterial load at 24 h, which was superior to vancomycin. In MRSA ATCC43300, their activity was similar to that of vancomycin. The high virulent CVCC546 strain displayed a relatively lower efficiency, with log CFU decreases of 2.88–2.91 (total), 3.41–3.50 (extracellular) and 2.11–2.51 (intracellular) compared with vancomycin (3.70). This suggests that MP1102/NZ2114 can be used as candidates for treating intracellular S. aureus.

Development of novel ultrashort antimicrobial peptide nanoparticles with potent antimicrobial and antibiofilm activities against multidrug-resistant bacteria.

Conventional antibiotics are facing strong microbial resistance that has recently reached critical levels. This situation is leading to significantly reduced therapeutic potential of a huge proportion of antimicrobial agents currently used in clinical settings. Antimicrobial peptides (AMPs) could provide the medical community with an alternative strategy to traditional antibiotics for combating microbial resistance. However, the development of AMPs into clinically useful antibiotics is hampered by their relatively low stability, toxicity, and high manufacturing costs. In this study, a novel in-house-designed potent ultrashort AMP named RBRBR was encapsulated into chitosan-based nanoparticles (CS-NPs) based on the ionotropic gelation method. The encapsulation efficacy reported for RBRBR into CS-NPs was 51.33%, with a loading capacity of 10.17%. The release kinetics of RBRBR from the nanocarrier exhibited slow release followed by progressive linear release for 14 days. The antibacterial kinetics of RBRBR-CS-NPs was tested against four strains of Staphylococcus aureus for 4 days, and the developed RBRBR-CS-NPs exhibited a 3-log decrease in the number of colonies when compared to CS-NP and a 5-log decrease when compared to control bacteria. The encapsulated peptide NP formulation managed to limit the toxicity of the free peptide against both mammalian cells and human erythrocytes. Additionally, the peptide NPs demonstrated up to 98% inhibition of biofilm formation when tested against biofilm-forming bacteria. Loading RBRBR into CS-NPs could represent an innovative approach to develop delivery systems based on NP technology for achieving potent antimicrobial effects against multidrug-resistant and biofilm-forming bacteria, with negligible systemic toxicity and reduced synthetic costs, thereby overcoming the obstructions to clinical development of AMPs.

CureCuma–cationic curcuminoids with improved properties and enhanced antimicrobial photodynamic activity

The naturally occurring photosensitizer curcumin has excellent biocompatibility, but its antimicrobial photodynamic efficacy is limited by (i) weak adherence to Gram(-) bacteria cell walls, (ii) low (photo-)stability and (iii) limited solubility in water.In this study novel curcuminoids bearing cationic substituents were prepared by different synthetic routes. The derivatives exhibit excellent water solubility, improved photostability and low aggregation.All novel curcuminoids showed antibacterial photodynamic effects (>3 log10 reduction of CFU) against Escherichia coli and Staphylococcus aureus upon blue light illumination. In contrast to natural curcumin, effective photokilling of E. coli was possible without the addition of permeabilizing agents. Ten micromolar of the most active compound (8) achieved a 7 log10 decrease of E. coli after light activation with a fluence of 33.8 J/cm2, whereas S. aureus was inactivated by more than 4 log10 at a fluence of 5.3 J/cm2. Overall the reduction in bacterial count was at least 100-fold more effective with these new curcuminoids in comparison to natural curcumin.

Two-phase kinetics of biofilm removal during CIP. Respective roles of mechanical and chemical effects on the detachment of single cells vs cell clusters from a Pseudomonas fluorescens biofilm

The objective of this study was to investigate the roles of mechanical and chemical actions on the detachment kinetics during CIP of biofilms of Pseudomonas fluorescens at a pilot-plant scale.Biofilms were grown 48 h at 20 °C in diluted milk. Shear stress conditions ranged from 0 (static) to 20 Pa and NaOH concentrations from 0.1 to 0.5% w/w. A simple two-phase model was applied to model the removal kinetics and compare detachment parameters.A quick removal of the biofilm during the first phase (4–5 log decrease in three minutes) was followed by a second phase of slow removal of the cells and small aggregates still present at the surface (0–2 log decrease). Hydrodynamics was responsible for removal of both biofilm and single cells while chemicals mainly disrupted biofilm clusters during the first phase. No complete biofilm removal was observed, suggesting a significant role of the interaction forces between bacteria and substrata in the CIP efficiency.

Activity of innate antimicrobial peptides and ivacaftor against clinical cystic fibrosis respiratory pathogens

There is a clear need for new antimicrobials to improve current treatment of chronic lung infection in people with cystic fibrosis (CF). This study determined the activities of antimicrobial peptides (AMPs) and ivacaftor, a novel CF transmembrane conductance regulator potentiator, for CF treatment. Antimicrobial activities of AMPs [LL37, human β-defensins (HβD) 1–4 and SLPI] and ivacaftor against clinical respiratory isolates (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus spp., Achromobacter spp. and Stenotrophomonas maltophilia) were determined using radial diffusion and time–kill assays, respectively. Synergy of LL37 and ivacaftor with tobramycin was determined by time–kill, with in vivo activity of ivacaftor and tobramycin compared using a murine infection model. LL37 and HβD3 were the most active AMPs tested, with MICs ranging from 3.2– ≥ 200 mg/L and 4.8– ≥ 200 mg/L, respectively, except for Achromobacter that was resistant. HβD1 and SLPI demonstrated no antimicrobial activity. LL37 demonstrated synergy with tobramycin against 4/5 S. aureus and 2/5 Streptococcus spp. isolates. Ivacaftor demonstrated bactericidal activity against Streptococcus spp. (mean log10 decrease 3.31 CFU/mL) and bacteriostatic activity against S. aureus (mean log10 change 0.13 CFU/mL), but no activity against other genera. Moreover, ivacaftor demonstrated synergy with tobramycin, with mean log10 decreases of 5.72 CFU/mL and 5.53 CFU/mL at 24 h for S. aureus and Streptococcus spp., respectively. Ivacaftor demonstrated immunomodulatory but no antimicrobial activity in a P. aeruginosa in vivo murine infection model. Following further modulation to enhance activity, AMPs and ivacaftor offer real potential as therapeutics to augment antibiotic therapy of respiratory infection in CF.

An Innovative Approach to Addressing the HIV Care Continuum: Implementation of a Clinical Pharmacy Resident in a Veterans Affairs HIV Specialty Clinic

Purpose: Engagement of patients in the HIV care continuum and adherence to antiretroviral therapy (ART) continue to limit successful viral suppression. Innovative practices to improve this continuum and ameliorate potential physician shortages are needed. The objective of this evaluation was to determine the clinical benefits of incorporating pharmacy resident involvement on a multidisciplinary team in caring for patients with HIV. Methods: A single-center pre–post cohort pilot evaluation was conducted at the Tennessee Valley Healthcare Systems VA Medical Center. Patients were enrolled in an HIV pharmacotherapy clinic implemented by an ambulatory care pharmacy resident. The primary end point of the evaluation was the percentage of patients achieving an undetectable plasma HIV viral load after enrollment. Secondary end points included change from baseline in CD4 T-cell count and self-reported adherence. Results: A total of 55 patients were seen in the HIV pharmacotherapy clinic over a 28-week evaluation period. Of those patients with detectable viral load at enrollment, 70% reached viral suppression during follow-up, with a significant 0.75 log10 decrease in the median viral load (P < .0001 for both). The median CD4 T-cell count increased from 464 to 525 cells/mm3 (P = .01). Reported adherence, assessed using the Visual Analogue adherence Scale (VAS) increased significantly (P = .0001). Conclusion: After enrollment in an HIV pharmacotherapy clinic, a significant decrease in viral load was seen, as were improvements in secondary end points of CD4 T cells and adherence. These data demonstrate the clinical benefits of pharmacy resident involvement on a multidisciplinary team in caring for patients with HIV.

In Vitro Activity of Colistin and Trimethoprim/Sulfamethoxazole Against Consortia of Multidrug Resistant Non-Fermenting Gram-Negative Bacilli Isolated from Lower Respiratory Tract

Background: Multidrug resistant (MDR) Pseudomonas aeruginosa, Acinetobacter baumannii and Stenotrophomonas maltophilia have a leading role in nosocomial infections, including lower respiratory tract (LRT) infections. When polymicrobial infection by these three bacteria occurs, colistin against MDR P. aeruginosa and A. baumannii and trimethoprim/sulfamethoxazole (SXT) against S. maltophilia can be an optional antimicrobial strategy. Objectives: The aim of this study was to investigate the potential synergic effect of colistin-plus-SXT against those MDR P. aeruginosa, A. baumannii and S. maltophilia isolates that were isolated at the same time, from the same LRT sample of patients. Methods: Sixty connected isolates from 20 different patients were collected in a two-year study period. The checkerboard method and time-kill assays were used for synergy testing. Results: All P. aeruginosa and A. baumannii strains were susceptible to colistin, whereas all S. maltophilia isolates were resistant to it. Fifteen percent of MDR A. baumannii strains and all S. maltophilia isolates were susceptible to SXT. By the checkerboard method, colistin-plus-SXT showed synergy in 50%, 35% and 45% of S. maltophilia, MDR P. aeruginosa and MDR A. baumannii strains, respectively. Antagonistic effect was not found. A time-kill assay was performed on strains which showed synergy by the checkerboard method: 70%, 57% and 56% of S. maltophilia, P. aeruginosa and A. baumannii strains showed the same results. Synergic activity of the combination was already detected after 6 h incubation in 86% of S. maltophilia isolates and 50% of P. aeruginosa strains. Regrowth of A. baumannii after 24 hour in the presence of colistin was prevented by the combination. The results gained by CB and TKA methods correlated in 61% of cases, but the ΣFIC values did not correlate with the rate of log10 decrease in TKA. Colistin-plus-SXT combination had synergic effect on 35% of S. maltophilia, 20% of P. aeruginosa and 25% A. baumannii strains by both methods. Conclusions: According to our in vitro results, colistin-plus-SXT combined therapy can be used efficiently in clinical practice as no antagonistic effect was detected. In certain cases colistin-plus-SXT has a synergic effect against MDR P. aeruginosa, A. baumannii and S. maltophilia.

Genetic and environmental factors influence Listeria monocytogenes nisin resistance.

Listeria monocytogenes nisin resistance increases when first exposed to NaCl and other stresses, such as low pH. In addition to environmental stressors, specific genomic elements can confer nisin resistance, such as the stress survival islet (SSI-1). As SSI-1 is variably present among L. monocytogenes strains, we wanted to determine if SSI-1 was associated with salt-induced nisin resistance. The presence of SSI-1 was determined using PCR for 48 strains of L. monocytogenes. When combined with multilocus sequence typing data, we found that the distribution of SSI-1 is clonal, where strains from clonal complexes (CC) 2, 6 and 11 do not have SSI-1, while strains from CCs 3, 5, 7 and 9 contain SSI-1. The impact of SSI-1 on salt-induced nisin resistance was dependent on CC. The average log decrease after 24h of exposure to nisin at 7°C under salt-inducing conditions was 2·6±1·1 for CC 9 strains and 2·3±0·7 for CC 11 strains, which had significantly lower survival compared to the other CCs, such as 1·3±0·3 for CC 6. Deletion of SSI-1 from a CC 7 strain demonstrated the role SSI-1 plays in salt-induced nisin resistance, as the deletion mutant had lower resistance compared to the parent strain. These data suggest that inducible nisin resistance in L.monocytogenes can be influenced by environmental conditions as well as the genetic composition of the strain, which should be considered when selecting control measures for ready-to-eat foods. The foodborne pathogen L.monocytogenes can grow in suboptimal conditions, including low temperature and high osmolarity, which makes it a safety concern for ready-to-eat foods. When using antimicrobial peptide inhibitors such as nisin, it is important to understand how food components can impact antimicrobial resistance across the genetic diversity of L. monocytogenes.

Disinfection efficiencies of sage and spearmint essential oils against planktonic and biofilm Staphylococcus aureus cells in comparison with sodium hypochlorite

Staphylococcus aureus causes human infections and foodborne intoxications. This study explored the potential antibacterial actions of sage and spearmint essential oils (EOs) against both its planktonic and biofilm cells, in comparison with sodium hypochlorite (NaOCl), a commonly applied chemical sanitizer. Initially, the minimum inhibitory and bactericidal concentrations (MICs, MBCs) of each plant mixture were determined against planktonic cultures, following growth at 30°C for 24h. Stationary phase planktonic bacteria were then individually exposed for 6min to either each EO (applied at 1–2×MBC; 2.5–5%), or NaOCl (250–450ppm). These were also left to form biofilms on 96-well polystyrene microplates, at 30°C for 96h, with medium renewal at 48h, in the presence of 10 different concentrations of each EO, expanding from sub- to super-inhibitory for planktonic growth, and the minimum biofilm inhibitory concentrations (MBICs; >90% inhibition) of each plant mixture were calculated. Formed biofilms were finally exposed for 6min to either each EO (applied at 2–6×MBC; 5–15%), or NaOCl (7500–25,000ppm; applied either alone or in combination with each EO at 5%). Results showed that both EOs presented MIC and MBC equal to 1.25 and 2.5%, respectively. As expected, their application at their MIC and above significantly inhibited biofilm formation, while spearmint EO was still able to cause this at ½ of its MIC, with MBICs equal to 1.25 and 0.63% for sage and spearmint EOs, respectively. Alarmingly, the application of both EOs at 1/8 to 1/16 of their MIC further increased biofilm formation. Regarding biofilm disinfection experiments, the individual application of each EO against the pre-established sessile communities resulted in log decrease ranges of 0.8–3logCFU/cm2, while in the case of NaOCl application (either alone or combined with each EO), the observed reductions never exceeded 1.7logCFU/cm2. These last results highlight the great antimicrobial recalcitrance of biofilm communities, found here to be ca. 100 times more resistant to NaOCl compared to planktonic ones, and stress the urgent need for further research on alternative, adequate and safe disinfection strategies to control them in food processing and other environments.

Crop bioaccumulation and human exposure of perfluoroalkyl acids through multi-media transport from a mega fluorochemical industrial park, China

Significant quantities of perfluoroalkyl acids (PFAAs) are released to the environment from fluorochemical manufacturing processes through wastewater discharge and air emission in China, which may lead to human exposure and health risks through crop bioaccumulation from PFAAs-contaminated soil and irrigation water. This paper systematically studied the distribution and transport of PFAAs in agricultural soil, irrigation water and precipitation, followed by crop bioaccumulation and finally human exposure of PFAAs within a 10km radius around a mega-fluorochemical industrial park (FIP). Hotspots of contamination by PFAAs were found near the FIP and downstream of the effluent discharge point with the maximum concentrations of 641ng/g in agricultural soil, 480ng/g in wheat grain, 58.8ng/g in maize grain and 4,862ng/L in precipitation. As the distance increased from the FIP, PFAAs concentrations in all media showed a sharp initial decrease followed by a moderate decline. Elevated PFAA concentrations in soil and grains were still present within a radius of 10 km of the FIP. The soil contamination was associated with the presence of PFAAs in irrigation water and precipitation, and perfluorooctanoic acid (PFOA) was the dominant PFAA component in soil. However, due to bioaccumulation preference, short-chain perfluoroalkyl carboxylic acids (PFCAs), especially perfluorobutanoic acid (PFBA), became the major PFAA contaminants in grains of wheat and maize. The bioaccumulation factors (BAFs) for both grains showed a decrease with increasing chain length of PFAAs (approximately 0.5 log decrease per CF2 group). Compared to maize grain, wheat grain showed higher BAFs, possibly related to its higher protein content. The PFCA (C4-C8) concentrations (on a log10 basis) in agricultural soil and grain were found to show a linear positive correlation. Local human exposure of PFOA via the consumption of contaminated grains represents a health risk for local residents, especially for toddlers and children.

Clinical and prognostic value of spot urinary creatinine in chronic heart failure—An analysis from GISSI-HF

ObjectivesThis study aimed to identify patient characteristics associated with low urinary creatinine in morning spot urine and investigate its association with clinical outcome. BackgroundTwenty-four-hour creatinine excretion is an established marker of muscle mass in heart failure and other populations. Spot urine creatinine might be an easy obtainable, cheap marker of muscle wasting and prognosis in heart failure (HF) patients. MethodsSpot urine creatinine concentration was measured in 2130 patients included in the GISSI-HF trial. We evaluated the prognostic value of urinary creatinine and its relation with clinical variables. ResultsMedian spot urinary creatinine was 0.80 (IQR 0.50-1.10) g/L. Lower spot urinary creatinine was associated with older age, smaller height and weight, higher NYHA class, worse renal function and more frequent spironolactone and diuretic use (all P<.02). During a median follow-up of 2.8 years, 655 patients (31%) experienced the combined endpoint of all-cause mortality or HF hospitalization. Lower urinary creatinine was independently associated with an increased risk of all-cause mortality or HF hospitalization (HR, 1.59 [1.21-2.08] per log decrease, P=.001), and all-cause mortality (HR, 1.75 [1.25-2.45] per log decrease, P=.001). ConclusionLower urinary creatinine, measured in morning spot urine in patients with chronic HF, is associated with worse renal function, smaller body size, more severe HF and is independently associated with an increased risk of all-cause death and HF hospitalization.

A Fresh Shine onCystic Fibrosis Inhalation Therapy: Antimicrobial Synergy of Polymyxin B in Combination with Silver Nanoparticles.

This in vitro study aimed to investigate the synergistic antibacterial activity of polymyxin B in combination with 2 nm silver nanoparticles (NPs) against Gram-negative pathogens commonly isolated from the cystic fibrosis (CF) lung. The in vitro synergistic activity of polymyxin B with silver NPs was assessed using the checkerboard assay against polymyxinsusceptible and polymyxin-resistant Pseudomonas aeruginosa isolates from the lungs of CF patients. The combination was also examined against the Gram-negative species Haemophilus influenzae, Burkholderia cepacia, Burkholderia pseudomallei, Stenotrophomonas maltophilia, Klebsiella pneumoniae and Acinetobacter baumannii that are less common in the CF lung. The killing kinetics of the polymyxin B-silver NPs combinations was assessed against P. aeruginosa by static time-kill assays over 24 h. Polymyxin B and silver NPs alone were not active against polymyxin-resistant (MIC ≥4 mg/L) P. aeruginosa. Whereas, the combination of a clinically-relevant concentration of polymyxin B (2 mg/L) with silver NPs (4 mg/L) successfully inhibited the growth of polymyxin-resistant P. aeruginosa isolates from CF patients as demonstrated by ≥2 log10 decrease in bacterial count (CFU/mL) after 24 h. Treatment of P. aeruginosa cells with the combination induced cytosolic GFP release and an increase of cellular reactive oxygen species. In the nitrocefin assay, the combination displayed a membrane permeabilizing activity superior to each of the drugs alone. The combination of polymyxin B and silver NPs displays excellent synergistic activity against highly polymyxin-resistant P. aeruginosa and is potentially of considerable clinical utility for the treatment of problematic CF lung infections.

Kinome Profiling Identifies Druggable Targets for Novel Human Cytomegalovirus (HCMV) Antivirals

Human cytomegalovirus (HCMV) is a significant cause of disease in immune-compromised adults and immune naïve newborns. No vaccine exists to prevent HCMV infection, and current antiviral therapies have toxic side effects that limit the duration and intensity of their use. There is thus an urgent need for new strategies to treat HCMV infection. Repurposing existing drugs as antivirals is an attractive approach to limit the time and cost of new antiviral drug development. Virus-induced changes in infected cells are often driven by changes in cellular kinase activity, which led us to hypothesize that defining the complement of kinases (the kinome), whose abundance or expression is altered during infection would identify existing kinase inhibitors that could be repurposed as new antivirals. To this end, we applied a kinase capture technique, multiplexed kinase inhibitor bead-mass spectrometry (MIB-MS) kinome, to quantitatively measure perturbations in >240 cellular kinases simultaneously in cells infected with a laboratory-adapted (AD169) or clinical (TB40E) HCMV strain. MIB-MS profiling identified time-dependent increases and decreases in MIB binding of multiple kinases including cell cycle kinases, receptor tyrosine kinases, and mitotic kinases. Based on the kinome data, we tested the antiviral effects of kinase inhibitors and other compounds, several of which are in clinical use or development. Using a novel flow cytometry-based assay and a fluorescent reporter virus we identified three compounds that inhibited HCMV replication with IC50 values of <1 μm, and at doses that were not toxic to uninfected cells. The most potent inhibitor of HCMV replication was OTSSP167 (IC50 <1.2 nm), a MELK inhibitor, blocked HCMV early gene expression and viral DNA accumulation, resulting in a >3 log decrease in virus replication. These results show the utility of MIB-MS kinome profiling for identifying existing kinase inhibitors that can potentially be repurposed as novel antiviral drugs.

68 The bifidobacterium longum 35624® culture transits in high numbers through the human gut

Background Most probiotic products rely on in vitro laboratory tests to assess transit and lack data on survival through the human gastrointestinal tract (GIT). However, in vitro tests are not representative of the multiple physiological states of the gut. Aim To confirm transit of the Bifidobacterium longum35 624 strain, found in Alflorex®, in different formats in humans. Method 35 624 strain was administered in a capsule, straw, sachet or milk format to volunteers for 7 days at doses ranging from 1x107–1 × 1010 CFU/day (n=1–6) followed by a wash-out period. The transit of viable 35 624 strain through the human GIT was assessed from stool collected at various timepoints by bifidobacteria selective agar plates and/or typing representative colonies using strain-specific PCR. Results Independent of form, the 35 624 strain transited in humans to high numbers, ranging from 1 × 104 – 1 × 108 CFU/g of stool in a dose-dependent manner at Day 7. Furthermore, despite antibiotic use in a 10-year-old cystic fibrosis sufferer (250 mg Zithromax 3 times/week), the 35 624 strain transited well (≥1 × 106 CFU/g). Co-administration of the antibiotic and probiotic resulted in a 1 log decrease in levels recovered from stool compared to administration of the probiotic 12 hour post antibiotic. Conclusion Independent of delivery format, the 35 624® strain was detected in stool at day 7 in a viable form at levels greater than 1×104 CFU/g of stool. Transit of viable 35 624® strain in a patient with long term antibiotic use was confirmed demonstrating that the Zithromax did not affect the 35 624® viability under the conditions of the experiment.

Effect of indigenous interferon - alpha on Hepatitis B virus deoxyribonucleic acid level in hepatitis b e antigen-positive chronic Hepatitis B patients

&lt;p class="ABS"&gt;&lt;span class="ABS_Bold-Italic" lang="en-GB"&gt;Background and Objective:&lt;/span&gt; HBeAg-positive chronic hepatitis B patients have high serum HBV DNA level showing high viral replication. Goal of treatment of hepatitis B is to prevent cirrhosis, hepatic failure and hepatocellular carcinoma by serum alanine transaminase (ALT) normalization, decrease in serum hepatitis B virus (HBV) deoxyribonucleic acid (DNA) and loss in hepatitis B e antigen (HBeAg). Interferons (IFNs) have antiviral, anti-proliferative, and immunomodulatory effects. IFN-&lt;span class="Symbol_Bold"&gt;α&lt;/span&gt; is effective in suppressing HBV replication and in inducing remission of liver disease. &lt;span class="ABS_Bold-Italic" lang="en-GB"&gt;Materials and Methods:&lt;/span&gt; In this prospective, single treatment arm study, HBeAg-positive chronic hepatitis patients without decompensated liver disease were enrolled to receive indigenous recombinant IFN-&lt;span class="Symbol_Bold"&gt;α&lt;/span&gt; 2b in the dose of 5 MU daily for 6 days a week subcutaneously for 16 weeks. Quantitative HBV-DNA, HBeAg, and hepatitis B surface antigen (HBsAg) were assessed at baseline and at the end of treatment. ALT level assessment was done at baseline and during therapy at week 1, week 2, week 8, week 12, and week 16. &lt;span class="ABS_Bold-Italic" lang="en-GB"&gt;Res&lt;/span&gt;&lt;span class="ABS_Bold-Italic" lang="en-GB"&gt;ults:&lt;/span&gt; Out of 37 patients enrolled in the study, 8 patients (21.62%) did not complete study due to lost to follow-up (3 patients), discontinuation due to adverse event (3 patients), and consent withdrawal (2 patients). Among 29 patients who completed the study, 10 patients (34.48%) had clearance of HBeAg and 1 patient (3.44%) had lost HBsAg after 16 weeks of therapy. Mean ALT level started decreasing after 4 weeks of therapy but did not come to normal range till 16 weeks of therapy. At least 2 log decreases in HBV DNA was observed in 9 (31.03%) patients and at least 1 log decrease in 18 (62.06%) patients. Overall decline in HBV DNA level was observed in 62% patients after 16 weeks of therapy. &lt;span class="ABS_Bold-Italic" lang="en-GB"&gt;Conclusion:&lt;/span&gt; IFN-&lt;span class="Symbol_Bold"&gt;α&lt;/span&gt; treatment does result in HBeAg and HBsAg loss and decreases HBV-DNA levels in chronic hepatitis B patients. Most of adverse events were mild to moderate in intensity. So, interferon-&lt;span class="Symbol_Bold"&gt;α&lt;/span&gt; therapy was well tolerated, safe, and efficacious to treat HBeAg-positive chronic hepatitis B patients without decompensated liver disease.&lt;/p&gt;

Survival of vaginal microorganisms in three commercially available transport systems

Transport systems are used to collect and maintain the viability of microorganisms. Two Amies media based transport systems, BD CultureSwab™ MaxV(+) Amies Medium without Charcoal (MaxV(+)) and Fisherfinest® with Amies gel Transport Medium without charcoal (Fisherfinest®) were compared to a Cary-Blair media based transport system, Starswab® Anaerobic Transport System (Starswab®), for their capacity to maintain the viability of 17 clinical microorganisms commonly isolated from the vagina (Lactobacillus crispatus, L. jensenii, L. iners, group B streptococci, Candida albicans, Escherichia coli, Enterococcus faecalis, Atopobium vaginae, Peptoniphilus harei, Mycoplasma hominis, Gardnerella vaginalis, Dialister microaerophilus, Mobiluncus curtisii, Prevotella amnii, P. timonensis, P. bivia, and Porphyromonas uenonis). Single swabs containing mixtures of up to five different species were inoculated in triplicate and held at 4 °C and room temperature for 24, 48, 72, and 96 h (h). At each time point, swabs were eluted into a sterile salt solution, serially diluted, inoculated onto selected media, and incubated. Each colony type was quantified and identified. A change in sample stability was reported as a ≥1 log increase or decrease in microorganism density from baseline. Overall, the viability of fastidious anaerobes was maintained better at 4 °C than room temperature. At 4 °C all three transport systems maintained the viability and prevented replication of C. albicans, E. faecalis, GBS, and E. coli. Microorganisms having a ≥1 log decrease in less than 24 h at 4 °C included A. vaginae, G. vaginalis, and P. uenonis in Starswab®, L. iners, A. vaginae, and P. amnii in MaxV(+), and A. vaginae, G. vaginalis, P. bivia, and P. amnii in Fisherfinest®. At 48 h at 4 °C, a ≥1 log decrease in concentration density was observed for P. harei and P. amnii in Starswab®, G. vaginalis, P. bivia and P. uenonis in MaxV(+), and L. iners, P. harei, P. timonensis, and P. uenonis in Fisherfinest®. Overall, at 4 °C the viability and stability of vaginal microorganisms was maintained better in the Cary-Blair based transport system (Starswab®) than in the two Amies based transport systems.

Synthesis and bactericidal properties of porphyrins immobilized in a polyacrylamide support: influence of metal complexation on photoactivity.

Spectroscopic and photodynamic properties of three novel polymeric hydrogels bearing porphyrins have been studied in vitro on the recombinant bioluminescent Gram-negative Escherichia coli DH5α to assess their ability to inactivate bacterial strains in solution. The three different hydrogels were formed by polymerization of 5-[4-2-(2-(2-acrylamidoethoxy)ethoxy)ethyl]carboxyphenyl-10,15,20-tris(4-N-methylpyridyl)porphyrin trichloride (5) and its complexes with Pd(ii) (6) and Cu(ii) (7) respectively, to form three optically transparent polyacrylamide hydrogels. All of the porphyrins are tricationic and they bear at the meso positions three N-methylpyridyl rings and one terminal acryloyl group connected through a flexible hydrophilic linker, particularly suitable for the later polymerization and incorporation into a hydrogel. The hydrogels were characterized by IR and scanning electron microscopy and incorporation of the dye was confirmed by UV-visible spectroscopy. All the hydrogels are characterized by a non-ordered microporous structure. The E. coli exhibited a decrease of 1.87 log after 25 min irradiation when the porphyrin hydrogel 9 was evaluated. When the Pd(ii) and Cu(ii)porphyrin hydrogels were tested (10, 11), they showed a 2.93 log decrease and 1.26 log decrease in the survival of the E. coli after 25 min irradiation, respectively. Similar results were obtained when the porphyrins were tested in solution. Of the three hydrogels, the Pd(ii)porphyrin hydrogel (10) proved to be the one with the highest photokilling ability under illumination, and also exhibited the lowest toxicity in the absence of light. Hydrogels 9 and 10 were found to be active for five cycles, suggesting the possibility of reuse.

From Breast Cancer to Antimicrobial: Combating Extremely Resistant Gram-Negative "Superbugs" Using Novel Combinations of Polymyxin B with Selective Estrogen Receptor Modulators.

Novel therapeutic approaches are urgently needed to combat nosocomial infections caused by extremely drug-resistant (XDR) "superbugs." This study aimed to investigate the synergistic antibacterial activity of polymyxin B in combination with selective estrogen receptor modulators (SERMs) against problematic Gram-negative pathogens. In vitro synergistic antibacterial activity of polymyxin B and the SERMs tamoxifen, raloxifene, and toremifene was assessed using the microdilution checkerboard and static time-kill assays against a panel of Gram-negative isolates. Polymyxin B and the SERMs were ineffective when used as monotherapy against polymyxin-resistant minimum inhibitory concentration ([MIC] ≥8 mg/L) Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii. However, when used in combination, clinically relevant concentrations of polymyxin B and SERMs displayed synergistic killing against the polymyxin-resistant P. aeruginosa, K. pneumoniae, and A. baumannii isolates as demonstrated by a ≥2-3 log10 decrease in bacterial count (CFU/ml) after 24 hours. The combination of polymyxin B with toremifene demonstrated very potent antibacterial activity against P. aeruginosa biofilms in an artificial sputum media assay. Moreover, polymyxin B combined with toremifene synergistically induced cytosolic green fluorescence protein release, cytoplasmic membrane depolarization, permeabilizing activity in a nitrocefin assay, and an increase of cellular reactive oxygen species from P. aeruginosa cells. In addition, scanning and transmission electron micrographs showed that polymyxin B in combination with toremifene causes distinctive damage to the outer membrane of P. aeruginosa cells, compared with treatments with each compound per se. In conclusion, the combination of polymyxin B and SERMs illustrated a synergistic activity against XDR Gram-negative pathogens, including highly polymyxin-resistant P. aeruginosa isolates, and represents a novel combination therapy strategy for the treatment of infections because of problematic XDR Gram-negative pathogens.

First Week HCV RNA Level Under the Pegylated Interferon and Ribavirin Treatment Predicts Sustained Virological Response

This study was planned to investigate whether the decrease in the hepatitis C virus (HCV) RNA levels at the first week of combined pegylated interferon and ribavirin treatment of naive genotype 1 patients with HCV was predicting sustained virologic response (SVR). Fifty-two patients were enrolled into the study. HCV RNA levels were measured at the baseline, first, fourth, and 12th weeks of treatment. Thirty-four patients achieved SVR, which basal, first week, and fourth week HCV RNA levels were log 5.57, log 3.65, and log 1.92, respectively. Eighteen patients could not achieve SVR, which basal, first week, and fourth week HCV RNA levels were log 6.22, log 5.45, and log 3.84, respectively (P < 0.05). Patients were distributed in 2 groups according to the amount of decrease in HCV RNA levels at the first week as less or more than 1.5 log. There were 20 patients with ≥1.5 log decrease in the HCV RNA levels at the first week. They were named as patients with very rapid virologic response (VRVR). All patients (100%) with VRVR were achieved SVR. In only 14 (44%) of the 32 patients without VRVR, SVR was achieved. In 16 (84%) of the 19 patients with rapid virologic response and 33 (79%) of the 42 patients with early virologic response, SVR was achieved. A ≥1.5 log decrease (VRVR) in HCV RNA levels of patients with HCV at the first week of combined pegylated interferon and ribavirin treatment predicts SVR very strongly.