Bacterial Penetration Research Articles

Nanostructures For Studying The Physics Of Biomolecules And Cells

I will argue that nanostructures open up new opportunities for biophysics and briefly present two examples: 1. We use fabricated nanopores to study DNA and DNA-protein constructs. 2. Micro/nanofabricated chambers and channels are used to study the motility and adaptive behavior of E. coli bacteria. I will report the limits of bacterial penetration through narrow fabricated constrictions.

Inhibitory effects of sucrose fatty acid esters on survival ofYersinia enterocoliticaon eggshell surface and use of blue lake as indicator of bacterial penetration into eggs

To determine the effectiveness of sucrose monolaurate (SML) and sucrose monocaprate (SMC), alone and in combination with ethylenediaminetetraacetic acid (EDTA), propionic acid (PA) or citric acid (CA) in reducing mesophilic aerobic bacteria (MAB) and Yersinia enterocolitica O:9 populations on eggshells and their damage potential on the microstructure of shell cuticle. Uninoculated eggs and eggs submerged in a solution of Y. enterocolitica were immersed in solutions of the various treatments. MAB and Y. enterocolitica counts on the surface of the eggs were carried out before and after treatment. MAB counts decreased less than 2 logs on uninoculated eggshells irrespective of treatment and reductions of 3.2 and 3.0 logs of Y. enterocolitica were obtained with 1000 microg ml(-1) SML plus 0.1% CA or 1000 microg ml(-1) SML plus 600 microg ml(-1) EDTA solutions, respectively. Y. enterocolitica 2/O:9 was recovered from natural microflora. Use of blue lake staining revealed minimal damage to the shells from the washing treatments. SML and SMC at 1000 microg ml(-1) combined with CA or EDTA could be effective in reducing Y. enterocolitica on eggshells with a minimal risk of later bacterial recontamination. Eggs are a recognized vehicle for transmission of Y enterocolitica although a prevalence of only 2.7% was detected in this study. Washing eggs in solutions containing SML or SMC could eliminate Y. enterocolitica contamination of egg shells.

Effect of phosphate buffer saline on coronal leakage of mineral trioxide aggregate

This study was carried out to compare the bacterial leakage of MTA used as a root-end filling material when it was kept in phosphate buffer saline (PBS) or normal saline. In this study, 72 freshly extracted teeth were used. The roots were randomly divided into four experimental groups of 15 each (groups I and II gutta-percha obturation + MTA, groups III and IV only MTA) and two positive and negative control groups of six each. The samples in groups I and III were kept in normal saline for 1 month while the samples in groups II and IV were kept in PBS. Enterococcus faecalis was used for determination of the bacterial penetration. Kaplan-Meier survival curve and chi(2) test were employed for data analysis. The obturated samples with root-end filling showed significantly longer duration of resistance to bacterial penetration than canals without obturation (P < 0.05). The roots that were placed in PBS (groups II and IV) showed significantly less bacterial penetration in comparison with the roots that were stored in normal saline (P < 0.05). In conclusion, MTA, which acts as a bioactive material, should be placed in a synthetic tissue fluid before any leakage evaluation.

Paneth cells directly sense gut commensals and maintain homeostasis at the intestinal host-microbial interface

The intestinal epithelium is in direct contact with a vast microbiota, yet little is known about how epithelial cells defend the host against the heavy bacterial load. To address this question we studied Paneth cells, a key small intestinal epithelial lineage. We found that Paneth cells directly sense enteric bacteria through cell-autonomous MyD88-dependent toll-like receptor (TLR) activation, triggering expression of multiple antimicrobial factors. Paneth cells were essential for controlling intestinal barrier penetration by commensal and pathogenic bacteria. Furthermore, Paneth cell-intrinsic MyD88 signaling limited bacterial penetration of host tissues, revealing a role for epithelial MyD88 in maintaining intestinal homeostasis. Our findings establish that gut epithelia actively sense enteric bacteria and play an essential role in maintaining host-microbial homeostasis at the mucosal interface.

The Effect of Age on Bacterial Penetration of Radicular Dentin

The aim of this study was to determine the effects of patient's age on the prevalence and depth of bacterial penetration inside dentinal tubules. Fifty-six single-rooted teeth were divided in two groups based on the patient's age: young (ages 18–25) and old (age ≥60). Teeth were instrumented and inoculated with a standardized bacterial load and incubated for 20 days. Histological analysis was performed to determine the degree of infection of the dentinal tubules by counting the number of invaded tubules and the depth of penetration of bacteria inside the tubules. A significantly higher number of tubules were invaded by bacteria in the young group compared with the old group (p = 0.014). Also, the depth of invasion by bacteria was significantly higher in the young than in the old group (p = 0.033). These results suggest that bacterial infection of dentinal tubules occur to a lesser extent in older patients.

Pathogen-dependent induction of cytokines and other soluble inflammatory mediators during intramammary infection of dairy cows1

Mastitis is a highly prevalent and costly disease of dairy cows that is commonly caused by intramammary bacterial infection. The innate immune response to bacterial penetration of the mammary gland is evoked within hours of infection, and the rapidity and magnitude of this response have been demonstrated to influence the resolution of this disease. Cytokines and other mediators of inflammation are known to play critical roles in the innate immune response to intramammary infection. The objectives of this review are to summarize the current understanding of the cytokine response to intramammary infection, highlight recent findings identifying differences in the cytokine response to various bacterial pathogens, and discuss future research directions that will increase our knowledge of the role of inflammatory mediators in predicting and governing the outcome of mastitis.

Chronic peripheral administration of corticotropin-releasing factor causes colonic barrier dysfunction similar to psychological stress

Chronic psychological stress causes intestinal barrier dysfunction and impairs host defense mechanisms mediated by corticotrophin-releasing factor (CRF) and mast cells; however, the exact pathways involved are unclear. Here we investigated the effect of chronic CRF administration on colonic permeability and ion transport functions in rats and the role of mast cells in maintaining the abnormalities. CRF was delivered over 12 days via osmotic minipumps implanted subcutaneously in wild-type (+/+) and mast cell-deficient (Ws/Ws) rats. Colonic segments were excised for ex vivo functional studies in Ussing chambers [short-circuit current (Isc), conductance (G), and macromolecular permeability (horseradish peroxidase flux)], and analysis of morphological changes (mast cell numbers and bacterial host-interactions) was determined by light and electron microscopy. Chronic CRF treatment resulted in colonic mucosal dysfunction with increased Isc, G, and horseradish peroxidase flux in+/+but not in Ws/Ws rats. Furthermore, CRF administration caused mast cell hyperplasia and abnormal bacterial attachment and/or penetration into the mucosa only in+/+rats. Finally, selective CRF agonist/antagonist studies revealed that stimulation of CRF-R1 and CRF-R2 receptors induced the elevated secretory state and permeability dysfunction, respectively. Chronic CRF causes colonic barrier dysfunction in rats, which is mediated, at least in part, via mast cells. This information may be useful in designing novel treatment strategies for stress-related gastrointestinal disorders.

Persistence of bacterial pathogens as a physiological phenomenon

The complex ecosystem of humans and microbes has been presented as an associative symbiosis based on a mutual support of symbionts with different consequences for them. Conditions for the persistence of bacteria have been defined: their resistance to environmental factors, antagonism in biocenosis, and stability to withstand host defense mechanisms. The key role of bacterial peptidoglycan has been defined for their survival in an infected organism, and the classification for persistent mechanisms of pathogens has been given. The group of bacterial secreted protease providing microbial resistance to defense factors of the organism has been described. Host colonizing resistance has been presented as a physiological regulatory system controlling bacterial penetration into an organism. Regulatory mechanisms for bacterial persistence have been discussed.

Development and Evaluation of Silver-Impregnated Amniotic Membrane as an Antimicrobial Burn Dressing

Silver has been widely used as an antimicrobial agent in burn wound care. A number of dressings containing silver have been developed using textiles, polyurethane films, foams, hydrogels, and hydrocolloids. However, biological-derived wound dressings have been advocated for their ability to more effectively promote healing and regulation of evaporation and exudation at the wound site. An antimicrobial burn dressing was developed by impregnation of silver on the amniotic membrane. The dressing was assessed for antimicrobial effectiveness and physical properties of relevance to clinical performance. Silver-impregnated dressings exhibited antimicrobial activity in contact for 5 days. An in vitro evaluation of the microbicidal efficacy of the silver-impregnated dressing was performed using Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans with different contamination level. More than 95% reduction in viable counts was observed in 2 to 4 hours. The release of silver from the dressings was observed for up to 4 days. Moisture vapor transmission rate was 1037 +/- 38 g/m2/24 hr for silver-impregnated dressings and 1024 +/- 44 g/m2/24 hr for amniotic membrane. The fluid absorption by the silver-impregnated dressings was comparable to the amniotic membranes. The total fluid handling capacity of the silver-impregnated amniotic membrane dressing examined ranged from 4 to 6.6 g/10 cm2 in 24 to 96 hours. The silver-impregnated amniotic membranes also provided an effective barrier to bacterial penetration. The study has demonstrated the ability of silver-impregnated amniotic membrane to combat microbial infection and its ideal physical characteristics for clinical use as a burn wound dressing.

Bacteria are not the primary cause of bleaching in the Mediterranean coral Oculina patagonica

Coral bleaching occurs when the endosymbiosis between corals and their symbionts disintegrates during stress. Mass coral bleaching events have increased over the past 20 years and are directly correlated with periods of warm sea temperatures. However, some hypotheses have suggested that reef-building corals bleach due to infection by bacterial pathogens. The 'Bacterial Bleaching' hypothesis is based on laboratory studies of the Mediterranean invading coral, Oculina patagonica, and has further generated conclusions such as the coral probiotic hypothesis and coral hologenome theory of evolution. We aimed to investigate the natural microbial ecology of O. patagonica during the annual bleaching using fluorescence in situ hybridization to map bacterial populations within the coral tissue layers, and found that the coral bleaches on the temperate rocky reefs of the Israeli coastline without the presence of Vibrio shiloi or bacterial penetration of its tissue layers. Bacterial communities were found associated with the endolithic layer of bleached coral regions, and a community dominance shift from an apparent cyanobacterial-dominated endolithic layer to an algal-dominated layer was found in bleached coral samples. While bacterial communities certainly play important roles in coral stasis and health, we suggest environmental stressors, such as those documented with reef-building corals, are the primary triggers leading to bleaching of O. patagonica and suggest that bacterial involvement in patterns of bleaching is that of opportunistic colonization.

Resistance of Disposable Drapes to Bacterial Penetration

To test the bacterial penetrability of disposable non-woven drapes used specifically for total hip arthroplasty. 12 round agar plates were inoculated with 107 colony-forming units/ml of coagulase-negative staphylococci (CNS) and incubated in air at 37 degrees Celsius for 18 hours to obtain a semi-confluent growth of organisms. Six brands of disposable drapes were tested; each was assigned to 2 plates. Each disposable drape was placed between a round agar plate and an inverted square agar plate filled with blood agar. After 30 and 90 minutes, the square agar plates were removed and incubated for 48 hours and inspected for growth of CNS. Bacterial penetration was time dependant. Certain brands of drapes were more impenetrable than others; none was impenetrable at all time points, but most remained so or allowed passage of fewer than 100 colony-forming units at 90 minutes. It is recommended that drapes be rigorously tested with regard to their resistance to bacterial penetration.

Bacterial leakage along the implant–abutment interface of premachined or cast components

In recent clinical studies, contamination of the inner parts of dental implants through bacterial penetration along the implant components has been observed. The aim of the present in-vitro study was to investigate leakage of Fusobacterium nucleatum through the interface between implants and premachined or cast abutments. Both premachined ( n = 10) and cast ( n = 10) implant–abutment assemblies were inoculated with 3.0 μL of microbial inoculum. The assemblies were completely immersed in 5.0 mL of tryptic soy broth culture medium to observe leakage at the implant–abutment interface after 14 days of anaerobic incubation. Bacterial growth in the medium, indicative of microbial leakage, was found only in 1 out of 9 samples (11.1%) in each group. Both premachined and cast abutments connected to external hexagonal implants provide low percentages of bacterial leakage through the interface in in vitro unloaded conditions if the manufacturer's instructions and casting procedures are properly followed.

Bacterial penetration along different root canal filling materials in the presence or absence of smear layer

To study the effect of the smear layer on the penetration of bacteria along different root canal filling materials and to examine the dentine/sealer and sealer/core material interfaces for the presence of bacteria. A total of 110 human root segments were instrumented to size 80 under irrigation with 1% sodium hypochlorite. Half of the roots were irrigated with a 5-mL rinse of 17% EDTA. Roots with and without smear layer were filled with gutta-percha (GP) and AH Plus sealer (AH), GP and Apexit sealer (AP), or RealSeal cones and sealer (RS). Following storage in humid conditions at 37 degrees C for 7 days, the specimens were mounted into a bacterial leakage test model for 135 days. Survival analyses were performed to calculate the median time of leakage and log-rank test was used for pairwise comparisons of groups. The level of significance was set at P = 0.05. Selected specimens were longitudinally sectioned and inspected by scanning electron microscopy for the presence of bacteria at the interfaces. In the presence of the smear layer, RS and AP leaked significantly more slowly than in its absence. In the absence of the smear layer, AH leaked significantly more slowly than RS. SEM results indicated a differential pattern of bacterial penetration among the sealers. Removal of the smear layer did not impair bacterial penetration along root canal fillings. A comparison of the sealers revealed no difference except that AH performed better than RS in the absence of the smear layer.

Responses of the pulp–dentin organ to dental restorative biomaterials

Placement of a restorative material in dentin produces the possibility of pulpal injury. In vitro studies have shown that the constituents of dental biomaterials have toxic potentials. In the clinical setting, there may be an immediate reaction of the pulp to, for example, acid etching and to the placement of a bonding agent; however, in most cases the remaining dentin serves as protection against long‐term or permanent damage to the pulp. Important factors for long‐term pulpal outcome are microleakage with possible bacterial penetration and leakage products from restorative materials. Both factors are influenced by the cavity depth, i.e., the remaining thickness of sound dentin. Received 13 February 2009; accepted 29 August 2009.

Unilaterale Halsschwellung bei Psoriasis

Local inflammation and systemic symptoms such as fever, fatigue and loss of appetite suggest the diagnosis of an abscess even if the patient is in a good general physical condition. The site of bacterial penetration should be identified along the corresponding lymphatic vessels. In this case, periauricular scratching induced by severe psoriasis led to a cervical abscess, the causing bacteria were identified as Streptococcus pyogenes.

Dentin structure in familial hypophosphatemic rickets: benefits of vitamin D and phosphate treatment

To evaluate the outcome of 1-(OH) vitamin D and oral phosphate treatment on dentin structure in patients with familial hypophosphatemic rickets, and expression of SIBLINGs (a family of non-collagenous proteins involved in dentinogenesis) and osteocalcin. Seven patients with familial hypophosphatemic rickets (age 3-16 years) were studied before or during treatment. Deciduous and permanent teeth were prepared for scanning electron microscopy (SEM) analysis and immunohistochemistry. Untreated or inadequately treated patients had necrotic teeth with impaired dentin mineralization including unmerged calcospherites and accumulation of non-collagenous proteins in wide interglobular spaces. Most of the primary incisors analyzed displayed fissures linking enamel subsurface to pulp horn. These elements may explain the bacterial penetration and dental abscesses despite the absence of carious lesions. Well-treated patients had healthy teeth with good dentin mineralization and little evidence of calcospherites. Treatment of hypophosphatemic children with 1-(OH) vitamin D and oral phosphate insures good dentin development and mineralization, and prevents clinical anomalies such as the dental necrosis classically associated with the disease. Starting treatment during early childhood and good adherence to the therapy are mandatory to observe these beneficial effects.

Septic osteitis of the distal phalanx in foals: 22 cases (1995–2002)

To determine the clinical characteristics and outcome of foals with septic osteitis of the distal phalanx. Retrospective case series. 22 foals. Information obtained from medical records included signalment; clinical, laboratory, and radiographic findings; treatment method; and outcome. Foals included in the study had lameness referable to the foot, radiographic evidence of localized lysis or focal loss of bone density of the distal phalanx, and suppurative discharge or necrosis of the affected bone evident at surgery. Foals with a history or evidence of penetrating wounds or subsolar abscessation were excluded. Mean age of foals at initial evaluation was 40.8 days (range, 3 to 122 days). Twenty-one (95%) foals had lameness as the primary complaint. Lesions consistent with septic osteitis of the distal phalanx localized to specific areas of the bone on the basis of radiographic and surgical findings were located on the solar margin or toe (14/22 [64%]), extensor process (5/22 [23%]), and palmar or plantar process (3/22 [13%]). Hind limbs (18/26 [69%] affected limbs) were more frequently affected. Two foals had > 1 affected limb, 2 had additional sites of osteomyelitis, and 4 had concurrent septic arthritis. Surgical debridement and regional antimicrobial perfusion were performed during general anesthesia. Extensor process lesions were not debrided. Nineteen of 22 (86%) foals survived to be discharged from hospital, and 16 horses reached racing age. Eleven of 16 had race starts, of which 8 had official race starts and 3 had unofficial race starts. Septic osteitis of the distal phalanx should be considered as a source of lameness in foals with signs referable to the foot and does not necessarily preclude a career in racing. Although infection may occur secondary to bacterial penetration of the hoof or sole, the distal phalanx should also be considered as a potential site for hematogenous septic arthritis or osteomyelitis in foals.

This Month in Gastroenterology

This Month in Gastroenterology

Efficacy of Two Contemporary Single-cone Filling Techniques in Preventing Bacterial Leakage

This in vitro study evaluated the sealing efficacy of three root-filling systems/techniques in preventing bacterial leakage. Instrumented single-rooted root segments were filled with (1) warm vertical compaction with gutta-percha/AH Plus; (2) single-cone technique with ActiV GP; and (3) single-cone technique with Gutta-Flow. A dual-chamber leakage model using S. mutans as a microbial marker was used for leakage evaluation. Bacterial penetration was monitored over a 100-day period. Leakage was recorded when turbidity was observed in the lower chamber. Gutta-percha warm vertical compaction exhibited the best seal with bacterial leakage observed in only 16.7% of the specimens between 59 and 100 days. All ActiV GP specimens leaked between 7 and 100 days; 50% of the Gutta-Flow specimens leaked between 22 and 100 days. The two contemporary single-cone techniques did not insure a durable apical seal against bacterial leakage. A warm vertical compaction technique using thermoplasticized gutta-percha and AH Plus sealer appears to be more effective in minimizing bacterial leakage.

Intracellular, Intercellular, and Stromal Invasion of Gastric Mucosa, Preneoplastic Lesions, and Cancer by Helicobacter pylori

It is not clear how Helicobacter pylori, an apparently extracellular pathogen colonizing the luminal side of the gastric epithelium, invariably causes an immune-inflammatory response on the stromal side of the mucosa. Penetration of H pylori into epithelial cell lines and its interaction with immune-inflammatory cells have been documented in vitro. Several investigations also showed in vivo bacterial penetration into the epithelium up to the lamina propria; however, the identification as H pylori of the bacteria-like bodies observed in unchanged, metaplastic, or neoplastic mucosa remained sometimes questionable. To search for bacteria-like organisms, we used transmission electron microscopy on endoscopic biopsy specimens from 20 dyspeptic subjects and surgical specimens of neoplastic and nonneoplastic mucosa from 20 cancerous stomachs. To ascertain the H pylori nature of the organisms found, we used 6 different antibodies directed against bacterial lysates, purified vacuolating cytotoxin A, or purified cytotoxin-associated antigen A in immunogold tests. The results were compared with those of H pylori strains cultivated in vitro. In nonmetaplastic gastric epithelium, cytochemically proven H pylori were detected, in the majority of cases, inside cytoplasm of epithelial cells, in intraepithelial intercellular spaces, and in underlying lamina propria, often in direct contact with immune-inflammatory cells and sometimes inside small blood vessels. Cytochemically proven H pylori were also observed inside 6 of 8 intestinal metaplasias and 9 of 20 cancers. H pylori penetrates normal, metaplastic, and neoplastic gastric epithelium in vivo, intracellularly, or interstitially to cause a strong immune-inflammatory response and promote gastric carcinogenesis.