Relationship between Biofilm-Formation, Phenotypic Virulence Factors and Antibiotic Resistance in Environmental Pseudomonas aeruginosa

The formation of a protective biofilm by Pseudomonas aeruginosa (PA) is one of the hallmarks of their survival both in vivo and in harsh environmental conditions, thus, biofilm-eradication has relevance from therapeutic perspectives and for infection control. The aim of our study was to investigate the possible relationship between antibiotic resistance, biofilm-forming capacity and virulence factors in n = 166 PA isolates of environmental origin. Antimicrobial susceptibility testing and the phenotypic detection of resistance determinants were carried out using standard protocols. The biofilm-forming capacity of PA was tested using a standardized crystal violet microtiter plate-based method. Motility (swimming, swarming, and twitching) and siderophore production of the isolates were also assessed. Resistance rates were highest for ciprofloxacin (46.98%), levofloxacin (45.18%), ceftazidime (31.92%) and cefepime (30.12%); 19.28% of isolates met the criteria to be classified as multidrug-resistant (MDR). Efflux pump overexpression, AmpC overexpression, and modified Hodge-test positivity were noted in 28.31%, 18.07% and 3.61%, respectively. 22.89% of isolates were weak/non-biofilm producers, while 27.71% and 49.40% were moderate and strong biofilm producers, respectively. Based on MDR status of the isolates, no significant differences in biofilm-production were shown among environmental PA (non-MDR OD570 [mean ± SD]: 0.416 ± 0.167 vs. MDR OD570: 0.399 ± 0.192; p > 0.05). No significant association was observed between either motility types in the context of drug resistance or biofilm-forming capacity (p > 0.05). 83.13% of isolates tested were positive for siderophore production. The importance of PA as a pathogen in chronic and healthcare-associated infections has been described extensively, while there is increasing awareness of PA as an environmental agent in agriculture and aquaculture. Additional studies in this field would be an important undertaking to understand the interrelated nature of biofilm production and antimicrobial resistance, as these insights may become relevant bases for developing novel therapeutics and eradication strategies against PA.

Awareness of Chronic Kidney Disease, Medication, and Laboratory Investigation among Nephrology and Urology Patients of Quetta, Pakistan.

Patients' awareness is critical in medical care, as it can serve as an input into the adjustment of interventions. The aim of study was to explore the level of awareness regarding chronic kidney disease (CKD), its medications, and laboratory investigations among nephrology and urology patients of Quetta. The cross-sectional study was used by adopting and culturally modifying a questionnaire. By convenient sampling technique, a total of 500 questionnaires were self-administered to inpatients, outpatients, and dialysis patients, and 468 responses (response rate 93.6%) were analyzed. Descriptive statistics, inferential statistics, and reliability analysis were performed on SPSS v25. A majority, 50.3% (n = 235), was unaware of symptoms that will develop due to worsening of disease, while 56.2% (n = 263) were unaware of what aggravates their kidney function. Almost half of the affected individuals, 47.4% (n = 222), have no understanding about the long-term prognosis of the disease. The majority of the respondents, 51.5% (n = 248), do not know about the names and usage of medications, and 62.4% (n = 292) were unaware of the medicines that may impair kidney function; more than half, 66.7% (n = 312), were unaware about the necessary laboratory investigations. A strong association between awareness and patient education level was found (p < 0.001). Awareness regarding disease condition, medications, and laboratory investigations of CKD among nephrology and urology patients of Quetta was found out to be low, which needs immediate educational intervention.

No Correlation between Biofilm-Forming Capacity and Antibiotic Resistance in Environmental Staphylococcus spp.: In Vitro Results.

The production of biofilms is a critical factor in facilitating the survival of Staphylococcus spp. in vivo and in protecting against various environmental noxa. The possible relationship between the antibiotic-resistant phenotype and biofilm-forming capacity has raised considerable interest. The purpose of the study was to assess the interdependence between biofilm-forming capacity and the antibiotic-resistant phenotype in 299 Staphylococcus spp. (S. aureus n = 143, non-aureus staphylococci [NAS] n = 156) of environmental origin. Antimicrobial susceptibility testing and detection of methicillin resistance (MR) was performed. The capacity of isolates to produce biofilms was assessed using Congo red agar (CRA) plates and a crystal violet microtiter-plate-based (CV-MTP) method. MR was identified in 46.9% of S. aureus and 53.8% of NAS isolates (p > 0.05), with resistance to most commonly used drugs being significantly higher in MR isolates compared to methicillin-susceptible isolates. Resistance rates were highest for clindamycin (57.9%), erythromycin (52.2%) and trimethoprim-sulfamethoxazole (51.1%), while susceptibility was retained for most last-resort drugs. Based on the CRA plates, biofilm was produced by 30.8% of S. aureus and 44.9% of NAS (p = 0.014), while based on the CV-MTP method, 51.7% of S. aureus and 62.8% of NAS were identified as strong biofilm producers, respectively (mean OD570 values: S. aureus: 0.779±0.471 vs. NAS: 1.053±0.551; p < 0.001). No significant differences in biofilm formation were observed based on MR (susceptible: 0.824 ± 0.325 vs. resistant: 0.896 ± 0.367; p = 0.101). However, pronounced differences in biofilm formation were identified based on rifampicin susceptibility (S: 0.784 ± 0.281 vs. R: 1.239 ± 0.286; p = 0.011). The mechanistic understanding of the mechanisms Staphylococcus spp. use to withstand harsh environmental and in vivo conditions is crucial to appropriately address the therapy and eradication of these pathogens.

Relationship between the Biofilm-Forming Capacity and Antimicrobial Resistance in Clinical Acinetobacter baumannii Isolates: Results from a Laboratory-Based In Vitro Study.

The relationship between the multidrug-resistant (MDR) phenotype and biofilm-forming capacity has been a topic of extensive interest among biomedical scientists, as these two factors may have significant influence on the outcomes of infections. The aim of the present study was to establish a possible relationship between biofilm-forming capacity and the antibiotic-resistant phenotype in clinical Acinetobacter baumannii (A. baumannii) isolates. A total of n = 309 isolates were included in this study. Antimicrobial susceptibility testing and the phenotypic detection of resistance determinants were carried out. The capacity of isolates to produce biofilms was assessed using a crystal violet microtiter-plate-based method. Resistance rates were highest for ciprofloxacin (71.19%; n = 220), levofloxacin (n = 68.61%; n = 212), and trimethoprim-sulfamethoxazole (n = 66.02%; n = 209); 42.72% (n = 132) of isolates were classified as MDR; 22.65% (n = 70) of tested isolates were positive in the modified Hodge-test; the overexpression of efflux pumps had significant effects on the susceptibilities of meropenem, gentamicin, and ciprofloxacin in 14.24% (n = 44), 6.05% (n = 19), and 27.51% (n = 85), respectively; 9.39% (n = 29), 12.29% (n = 38), 22.97% (n = 71), and 55.35% (n = 170) of isolates were non-biofilm-producing and weak, moderate, and strong biofilm producers, respectively. A numerical, but statistically not significant, difference was identified between the MDR and non-MDR isolates regarding their biofilm-forming capacity (MDR: 0.495 ± 0.309 vs. non-MDR: 0.545 ± 0.283; p = 0.072), and no association was seen between resistance to individual antibiotics and biofilm formation. Based on numerical trends, MER-resistant isolates were the strongest biofilm producers (p = 0.067). Our study emphasizes the need for additional experiments to assess the role biofilms have in the pathogenesis of A. baumannii infections.

Association between biofilm-production and antibiotic resistance in Escherichia coli isolates: A laboratory-based case study and a literature review.

Bacteria can enhance their survival by attaching to inanimate surfaces or tissues, and presenting as multicellular communities encased in a protective extracellular matrix called biofilm. There has been pronounced interest in assessing the relationship between the antibiotic resistant phenotype and biofilm-production in clinically-relevant pathogens. The aim of the present paper was to provide additional experimental results on the topic, testing the biofilm-forming capacity of Escherichia coli isolates using in vitro methods in the context of their antibiotic resistance in the form of a laboratory case study, in addition to provide a comprehensive review of the subject. In our case study, a total of two hundred and fifty (n = 250) E. coli isolates, originating from either clean-catch urine samples (n = 125) or invasive samples (n = 125) were included. The colony morphology of isolates were recorded after 24h, while antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. Biofilm-formation of the isolates was assessed with the crystal violet tube-adherence method. Altogether 57 isolates (22.8%) isolates were multidrug resistant (MDR), 89 isolates (35.6%) produced large colonies (>3 mm), mucoid variant colonies were produced in 131 cases (52.4%), and 108 (43.2%) were positive for biofilm formation. Biofilm-producers were less common among isolates resistant to third-generation cephalosporins and trimethoprim-sulfamethoxazole (P = 0.043 and P = 0.023, respectively). Biofilms facilitate a protective growth strategy in bacteria, ensuring safety against environmental stressors, components of the immune system and noxious chemical agents. Being an integral part of bacterial physiology, biofilm-formation is interdependent with the expression of other virulence factors (especially adhesins) and quorum sensing signal molecules. More research is required to allow for the full understanding of the interplay between the MDR phenotype and biofilm-production, which will facilitate the development of novel therapeutic strategies.

Antimicrobial Resistance in the Context of the Sustainable Development Goals: A Brief Review.

The reduction in infectious disease morbidity and mortality may be attributed to a variety of factors; however, improved sanitation and public health, and the introduction of vaccines and antibiotics are among the most significant. The development of antimicrobial resistance (AMR) in bacterial pathogens is an expected consequence of evolutionary adaptation to these noxious agents and the widespread use of these drugs has significantly sped up this process. Infections caused by multidrug resistant pathogens are directly associated with worse clinical outcomes, longer hospital stays, excess mortality in the affected patients and an increasing burden and costs on the healthcare infrastructure. The Sustainable Development Goals (SDGs) were published in 2015 by the United Nations to serve as a global blueprint for a better, more equitable, more sustainable life on our planet. The SDGs contextualize AMR as a global public health and societal issue; in addition, the continuing emergence of AMR may limit the attainment on many SDGs. The aim of this mini-review is to provide insight on the interface between attainment of SDGs and the clinical problem of drug resistance in bacteria.

Correlation Between Biofilm-Formation and the Antibiotic Resistant Phenotype in Staphylococcus aureus Isolates: A Laboratory-Based Study in Hungary and a Review of the Literature.

INTRODUCTION: Staphylococcus aureus (S. aureus) is an important causative pathogen in human infections. The production of biofilms by bacteria is an important factor, leading to treatment failures. There has been significant interest in assessing the possible relationship between the multidrug-resistant (MDR) status and the biofilm-producer phenotype in bacteria. The aim of our present study was to assess the biofilm-production rates in clinical methicillin-susceptible S. aureus [MSSA] and methicillin-resistant S. aureus [MRSA] isolates from Hungarian hospitals and the correlation between resistance characteristics and their biofilm-forming capacity. METHODS: A total of three hundred (n=300) S. aureus isolates (corresponding to MSSA and MRSA isolates in equal measure) were included in this study. Identification of the isolates was carried out using the VITEK 2 ID/AST system and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method and E-tests, confirmation of MRSA status was carried out using PBP2a agglutination assay. Biofilm-production was assessed using the crystal violet (CV) tube-adherence method and the Congo red agar (CRA) plate method. RESULTS: There were significant differences among MSSA and MRSA isolates regarding susceptibility-levels to commonly used antibiotics (in case of erythromycin, clindamycin and ciprofloxacin: p<0.001, gentamicin: p=0.023, sulfamethoxazole/trimethoprim: p=0.027, rifampin: p=0.037). In the CV tube adherence-assay, 37% (n=56) of MSSA and 39% (n=58) of MRSA isolates were positive for biofilm-production, while during the use of CRA plates, 41% (n=61) of MSSA and 44% (n=66) of MRSA were positive; no associations were found between methicillin-resistance and biofilm-production. On the other hand, erythromycin, clindamycin and rifampin resistance was associated with biofilm-positivity (p=0.004, p<0.001 and p<0.001, respectively). Biofilm-positive isolates were most common from catheter-associated infections. DISCUSSION: Our study emphasizes the need for additional experiments to assess the role biofilms have in the pathogenesis of implant-associated and chronic S. aureus infections.

Small, but smelly: the importance of Solobacterium moorei in halitosis and other human infections.

Solobacterium moorei (S. moorei) has been described as Gram-positive, non spore forming, obligate anaerobic bacillus from human feces. The traditional culture and identification of these strains is very difficult (as the strains are often not cultivable or they grow only relatively slowly, in addition to producing only a very few positive biochemical reactions in commercially available identification kits); thus, reliable identification may only be carried out using methods, such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and DNA sequencing. Regarding its pathogenic role, the relevance of S. moorei in halitosis (oral malodor) has a good standing, as it has been suggested by multiple studies, while the isolation of these bacteria from invasive infections is very rare; there are only a few reports available in the literature, regarding infections outside the oral cavity. Based on these reports, affected patients are predominantly characterized compromised immunity and are frequently associated with a dental focus of infection. The aim of our present review is to summarize the currently available knowledge on the pathogenic role of S. moorei in halitosis and other infections and to emphasize the relevance of this neglected anaerobic pathogen.

Effects of different decontaminating solutions used for the treatment of peri-implantitis on the growth of Porphyromonas gingivalis-an in vitro study.

Implants have been considered the treatment of choice to replace missing teeth, unfortunately, peri-implant disease is still an unresolved issue. Contaminated implants may be decontaminated by physical debridement and chemical disinfectants; however, there is a lack of consensus regarding the ideal techniques/agents to be used for the decontamination. The objective of our study was to compare the decontaminating efficacy of different chemical agents on a titanium surface contaminated with Porphyromonas gingivalis, a typical representative of the bacterial flora associated with peri-implantitis. Commercially pure Ti grade 4 discs with a polished surface were treated with a mouthwash containing chlorhexidine digluconate (0.1%), povidone-iodine (PVP-iodine) solution (10%) or citric acid monohydrate (40%). Qualitative and quantitative assessment of cellular growth and survival were assessed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and scanning electron microscopy (SEM). Significant differences in the quantity of P. gingivalis could be observed after 6 days of incubation. A numerical, but not statistically significant (P = 0.066) decrease in the amount of living bacteria was observed in the group treated with the PVP-iodine solution as compared to the control group. The chlorhexidine (CHX)-treated group presented with significantly higher cell counts, as compared to the PVP-iodine-treated group (P = 0.032), while this was not observed compared to the control group and citric acid-treated group. Our results have also been verified by SEM measurements. Our results suggest that for P. gingivalis contamination on a titanium surface in vitro, PVP-iodine is a superior decontaminant, compared to citric acid and chlorhexidine-digulconate solution.

Streptococcus suis: An Underestimated Emerging Pathogen in Hungary?

Streptococcus suis (S. suis) is an emerging zoonotic pathogen, demonstrated as an etiological agent in human infections in increasing frequency, including diseases like purulent meningitis, sepsis, uveitis-endophtalmitis and arthritis. Due to the increased availability and utility of novel diagnostic technologies in clinical microbiology, more studies have been published on the epidemiology of S. suis, both in veterinary and human medicine; however, there are no comprehensive data available regarding human S. suis infections from East-Central European countries. As a part of our study, data were collected from the National Bacteriological Surveillance (NBS) system on patients who had at least one positive microbiological result for S. suis, corresponding to an 18-year study period (2002-2019). n = 74 S. suis strains were isolated from invasive human infections, corresponding to 34 patients. The number of affected patients was 1.89 ± 1.53/year (range: 0-5). Most isolates originated from blood culture (63.5%) and cerebrospinal fluid (18.9%) samples. Additionally, we present detailed documentation of three instructive cases from three regions of the country and with three distinctly different outcomes. Hungary has traditional agriculture, the significant portion of which includes the production and consumption of pork meat, with characteristic preparation and consumption customs and unfavorable epidemiological characteristics (alcohol consumption, prevalence of malignant diseases or diabetes), which have all been described as important predisposing factors for the development of serious infections. Clinicians and microbiologist need to be vigilant even in nonendemic areas, especially if the patients have a history of occupational hazards or having close contact with infected pigs.

A Clinicopathological Approach to Odontogenic Cysts: the Role of Cytokeratin 17 and bcl2 Immunohistochemistry in Identifying Odontogenic Keratocysts.

Odontogenic keratocysts (OKCs) are developmental cysts of the jaws that require proper diagnosis due to their potential for local aggressive growth and recurrences. OKCs have a typical parakeratotic epithelium demonstrating transepithelial cytokeratin 17 (CK17) and basal bcl2 staining on immunohistochemistry (IHC), which distinguishes them from other common jaw cysts. Secondary to inflammation, the epithelial lining may be altered and loses the typical IHC phenotype. The aim of the present study was to analyse a series of consecutive jaw cysts for their expression of CK17 and bcl2 and assess how these IHC stains may help in their diagnosis. All cysts were retrospectively assessed for available clinical, radiological and pathological findings and diagnoses were revised whenever needed. 85 cysts from 72 patients were collected from two departments. The series had 21 OKCs, the remaining non-OKCs included radicular/residual, dentigerous, paradental, lateral periodontal, botryoid odontogenic cysts. OKCs with typical epithelium showed the typical IHC phenotype, which was generally lost in inflammation-associated altered epithelium. Contrarily to earlier descriptions, a wide variety of CK17 positivity was seen in the majority of non-OKCs, including focal transepithelial staining. Basal bcl2 staining was also seen in 16 non-OKCs. These stainings were never as strong in intensity as seen in OKCs. One case was histopathologically identified as OKC due to focally maintained IHC profile. CK17 and bcl2 IHC may help in the diagnosis of OKCs, but must be interpreted with caution and is not a yes or no tool in the diagnostic puzzle.

Utility of Photodynamic Therapy in Dentistry: Current Concepts.

The significant growth in scientific and technological advancements within the field of dentistry has resulted in a wide range of novel treatment modalities for dentists to use. Photodynamic therapy (PDT) is an emerging, non-invasive treatment method, involving photosensitizers, light of a specific wavelength and the generation of singlet oxygen and reactive oxygen species (ROS) to eliminate unwanted eukaryotic cells (e.g., malignancies in the oral cavity) or pathogenic microorganisms. The aim of this review article is to summarize the history, general concepts, advantages and disadvantages of PDT and to provide examples for current indications of PDT in various subspecialties of dentistry (oral and maxillofacial surgery, oral medicine, endodontics, preventive dentistry, periodontology and implantology), in addition to presenting some images from our own experiences about the clinical success with PDT.

Diagnosis and Management of Cervicofacial Actinomycosis: Lessons from Two Distinct Clinical Cases.

Members of the Actinomyces genus are non-spore-forming, anaerobic, and aerotolerant Gram-positive bacteria that are abundantly found in the oropharynx. They are the causative agents of actinomycosis, a slowly progressing (indolent) infection with non-specific symptoms in its initial phase, and a clinical course of extensive tissue destruction if left untreated. Actinomycoses are considered to be rare; however, reliable epidemiological data on their prevalence is lacking. Herein, we describe two representative and contrasting cases of cervicofacial actinomycosis, where the affected patients had distinctively different backgrounds and medical histories. Identification of the relevant isolates was carried out using matrix-assisted laser desorption/ionization mass spectrometry; antimicrobial susceptibility was performed using E-tests. Cervicofacial actinomycoses are the most frequent form of the disease; isolation and identification of these microorganisms from relevant clinical samples (with or without histological examination) is the gold standard for diagnosis. The therapy of these infections includes surgical debridement and antibiotic therapy, mainly with a penicillin-derivative or clindamycin.

Immunohistochemical Characterization of Reactive Epithelial Changes in Odontogenic Keratocysts.

Odontogenic keratocysts (OKCs) have a diagnostic thin epithelial lining characterised by a linear epithelial connective tissue interface generally lacking inflammatory changes, basal palisading of the nuclei and a wavy parakeratotic layer on the surface. This typical epithelium may convert to a thicker non-keratinizing one with rete pegs and a relatively flat surface after operative decompression. The aim was to characterize this type of epithelial change by immunohistochemistry for bcl2, keratin17, 10 and 19. Eleven out of 33 archived OKCs demonstrated an altered epithelium related to previous biopsy, decompressing drainage or inflammation. The typical basal bcl2 staining was lost in 10/11 cases; transepithelial CK17 was lost or markedly reduced in 9/11 cases. CK10 displayed a segmental upper layer staining in OKCs, and its loss or partial loss in the altered epithelium did not differ from negative areas of OKCs. CK19 displayed various staining patterns in the altered epithelium from lost to maintained in a patchy transepithelial distribution, the latter of which did not differ from the typical OKC staining pattern. Three of four non-keratinizing epithelial linings with basal palisading displayed immunostaining reminiscent of typical OKC epithelium. The lack of a typical epithelium is not sufficient to exclude the diagnosis of OKC if the sampling is not generous (e.g. biopsy), and the presence of non-keratinizing epithelium with basal palisading and an immunophenotype characteristic of OKC (basal bcl2, patchy or diffuse CK17 and upper layer CK10 positivity) may be consistent with the OKC diagnosis even in the absence of typical epithelial lining.

Comparison of effects of different fluoride containing prophylactics on the growth of Streptococcus mutans on titanium surfaces - an in vitro study.

The combined use of high fluoride (F-) concentration and acidic pH can weaken the corrosion resistance of titanium (Ti). Caries prophylactic products contain high amounts of F- and are applied at a low pH. The aim of our study was to determine whether the different forms of applied flouride has different effects on the growth of Streptococcus mutans on different titanium surfaces. Titanium with polished surface were treated with a gel (pH: 4,8) containing 1,25% olaflur, a rinse (pH: 4,4) containing 0,025% olaflur or a 1% aqueous solution of NaF at a pH of 4,5. Control discs were not treated. All discs were incubated with S. mutans for 21 days. To assess the amount of S. mutans protein assay analysis was performed at 5, 10 and 21 days. Scanning electron microscopic (SEM) investigations were also executed. By the 21st day significant differences could be observed in the bacterial protein quantity. The between group- comparisons showed that the rinse and gel were superior to NaF or control group (p < 0,01 and p < 0,05). Furthermore signs of corrosion could be observed in the group of gel treated discs. The results suggest that amine-fluoride content mouthwashes might be a suitable choice for prevention to the patients with dental implants.

Short- and Long-Term Influence of Fluoride-Containing Prophylactics on the Growth of Streptococcus mutans on Titanium Surface.

OBJECTIVES: Acidic pH and high fluoride (F(-)) concentration impair the corrosion resistance of titanium (Ti). Caries-preventive products contain high amounts of F(-) and are applied at low pH. The purpose of this study was to evaluate whether fluoride applied in different forms has different short-, mid-, and long-term effects on the growth of the bacteria Streptococcus mutans. MATERIALS AND METHODS: Ti discs with polished surface were treated with a rinse containing 0.025% olaflur, a gel containing 1.25% olaflur, or a 1% aqueous solution of NaF (pH 4.5), and they were incubated with S mutans for 21 days. Control discs did not get prophylactic treatment. Protein assay analysis was performed at regular intervals to estimate the amount of S mutans. Scanning electron microscopic (SEM) images were also taken. RESULTS: Bacterial protein quantity became significantly different only by the 21st day. Fluoride in rinse and gel proved to be superior to NaF in aqueous solution or no treatment (P < 0.01 and P < 0.05, respectively). However, the discs treated with fluoride in gel showed signs of corrosion in SEM images. CONCLUSION: The results suggest that the use of fluoride-containing mouthwashes might be the best and safest oral hygienic choice for patients with oral implants. Furthermore, olaflur seems to be superior to NaF for long-term use at low pH.

Prevalence of exocrine pancreatic insufficiency in type 2 diabetes mellitus with poor glycemic control.

OBJECTIVES: To evaluate the relationship between exocrine pancreatic insufficiency and the level of glycemic control in diabetes (DM). METHODS: Patients with type 2 DM treated in our clinic were prospectively recruited into the study. Pancreatic diabetes was excluded. Cases with HbA1c ≥7% formed Group A (n = 59), and with HbA1c <7% Group B (n = 42). The fecal level of pancreatic elastase (PE-1) was measured and morphological examinations of the pancreas were performed. RESULTS: The PE-1 level was significantly lower in Group A than in Group B (385.9 ± 171.1 µg/g, vs. 454.6 ± 147.3 µg/g, p = 0.038). The PE-1 level was not correlated with HbA1c (r = -0.132, p = 0.187), the duration of DM (r = -0.046, p = 0.65), age (r = 0.010, p = 0.921), BMI (r = 0.203, p = 0.059), or pancreatic steatosis (r = 0.117, p = 0.244). The size of the pancreas did not differ significantly between Groups A and B. CONCLUSIONS: An exocrine pancreatic insufficiency demonstrated by fecal PE-1 determination is more frequent in type 2 DM patients with poor glycemic control. The impaired exocrine pancreatic function cannot be explained by an alteration in the size of the pancreas or by pancreatic steatosis.

Effect of caries preventive products on the growth of bacterial biofilm on titanium surface.

Fluorides may affect the oxide layer on titanium surface. Caries preventive mouthwashes or gels contain fluorides and are applied at low pH. The aim of the present work was to study whether various concentrations of fluoride at acidic pH cause changes in the surface structure on the polished region of Ti implants, and alter the adherence and colonization of bacteria. Commercially pure Ti grade 4 discs with a polished surface were treated with a mouthwash containing 0.025% fluoride, a gel containing 1.25% fluoride or a 1% aqueous solution of NaF (pH 4.5). The change of surface roughness of the samples and the colonization of Porphyromonas gingivalis strains were studied by scanning electron microscopy after 5 days of anaerobic incubation. The quantity of the bacterial protein was determined by protein assay analysis. Agents with high fluoride concentration at acidic pH increased the roughness of the Ti surface. A slight increase in the amount of bacteria was found on the surfaces treated with 1% NaF and gel in comparison with the control surface. This study suggested that a high fluoride concentration at acidic pH may hinder the development of a healthy transgingival epithelial junction on Ti implants, due to bacterial colonization.

[Streptococcus mutans colonization on titanium surfaces treated with various fluoride-containing preventive solutions]. / A Streptococcus mutans kolonizációja különbözô fluoridot tartalmazó prevenciós oldatokkal kezelt titánfelszínen.

Fluoride is a reductive agent and may modify the oxide layer of titanium (Ti) in the transgingival region of dental implants. The low pH and the high fluoride concentration of prophylactic mouthwashes and gels (used in caries prevention) may play a role in this phenomenon. Our main goal was to examine whether changes on the surface structure of Ti caused by high fluoride concentration and acidic pH alter the adherence and the colonization of bacteria. Polished commercially pure Ti discs (CP grade 4, Camlog, Biotechnologies AG, Switzerland) were used in the study. Each sample was treated for 1 hour with one of the solutions: mouthwash containing 0.025% (250 ppm) fluoride, a gel containing 1.25% (12500 ppm) fluoride, and a solution of 1% NaF (3800 ppm fluoride), pH 4.5. The surface structure of the discs was analyzed by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). The colonization of Streptococcus mutans was studied by scanning electron microscope (SEM) after a 5-day incubation period. The roughness of the treated sample surfaces (Ra), as revealed by AFM measurements, increased 1.3 times for the gel and the mouthwash, and approximately seven folds for the 1% NaF solution, as compared to the control surface. The high fluoride concentration and acidic pH of the gel and the 1% NaF solution resulted in a strong corrosion and a modification of the composition of the Ti surface. The XPS spectra showed the formation of a fluoride containing complex (Na2TiF6) bound strongly to the surface. A correlation was revealed between the roughness of the surface and thickness and maturity of the S. mutans bacterial colonies developed on the modified Ti surface. High fluoride concentration and acidic pH increased the roughness of the Ti surface. Bacterial biofilm colonization on this rough surface proved to be more mature. The amount of bacteria was increased due to the changes in the surface caused by fluoride treatment. The present study indicates that high fluoride concentration in an acidic pH environment may affect the development of a healthy transgingival epithelial junction on the Ti surface. This work was supported by the SIMI-NAS Project of the 5th FWP of the European Commission (Growth Program, GRD3-2001-61801), the Hungarian Ministry of Economy and the EC (GVOP-3.2.1.-2004-04-0408/3.0), the Hungarian Ministry of Health (ETT, 434/2006), and the Hungarian Scientific Research Fund (OTKA F-68440).

Corrosive effects of fluoride on titanium: investigation by X-ray photoelectron spectroscopy, atomic force microscopy, and human epithelial cell culturing.

High fluoride (F(-)) concentrations and acidic pH impair the corrosion resistance of titanium (Ti). Effects of F(-)-containing caries-preventive prophylactic rinses, and gels on Ti were investigated by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). Human epithelial cell attachment and proliferation were investigated by dimethylthiazol-diphenyl tetrazolium bromide (MTT) and protein content assays. Aqueous 1% NaF solution (3800 ppm F(-), pH 4.5) or high (12,500 ppm) F(-) content gel (pH 4.8) strongly corroded the surface and modified its composition. XPS revealed formation of a strongly bound F(-)-containing complex (Na(2)TiF(6)). AFM indicated an increase in roughness (R(a)) of the surfaces: 10-fold for the NaF solution and smaller for the gel or a mouthwash (250 ppm F(-), pH 4.4). MTT revealed that cell attachment was significantly increased by the gel, but was not disturbed by either the mouthwash or the NaF. Cell proliferation determined by MTT decreased significantly only for the NaF-treated samples; protein content assay experiments showed no such effect. This study indicates that epithelial cell culturing results can depend on the method used, and the adverse effects of a high F(-) concentration and low pH should be considered when prophylactic gels are applied by patients with Ti implants or other dental devices.